december 18 to 20, 2014 the chinese university of hong kong · table of contents the 2014...

December 18 to 20, 2014 The Chinese University of Hong Kong

HONG KONG, CHINA

18th – 20th December 2014

Final Programme and Information to Delegates

Table of Contents The 2014 International Conference on Photodynamic Therapy and Translational Medicine

Table of Contents

General Information .................................................................................................... ii

Welcome Message from the Organizer ....................................................................... iii

Committees ................................................................................................................ iv

Organizers and Sponsors.............................................................................................. v

Site Map ..................................................................................................................... vi

Programme at a Glance ............................................................................................. vii

Floor Plan .................................................................................................................. viii

Scientific Programme ................................................................................................... 1

Workshops ................................................................................................................... 6

List of Posters .............................................................................................................. 7

Keynote Speakers ........................................................................................................ 8

Abstracts – Plenary Lectures ........................................................................................ 9

Abstracts – PDT and Translational Medicine .............................................................. 13

Abstracts – PDT Application and Technologies .......................................................... 18

Abstracts – Poster Session ......................................................................................... 20

List of Participants ..................................................................................................... 30

Index of Authors ........................................................................................................ 32

The 2014 International Conference on Photodynamic Therapy and Translational Medicine General Information

ii

General Information CONFERENCE VENUE

Ground floor, Esther Lee Building, The Chinese University of

Hong Kong, Shatin, Hong Kong

Phone: +852 39434329

Email: [email protected]

OFFICIAL LANGUAGE

The official language of the conference is English.

REGISTRATION

Registration is in the lobby of the ground floor of the

conference venue during the following hours.

18th

December 2014: 15:00 – 20:00

19th

December 2014: 08:00 – 18:00

20th

December 2014: 08:00 – 18:00

CONFERENCE BADGE

Conference badges must be worn all the times while

delegates are on the conference venue. It is for admittance

to all events.

POSTER PRESENTATION

All poster presentations are on the ground floor of the

conference venue. Presenters should place their posters on

the designated poster board indicated in the conference

program. Presenters may set up and take down their posters

during the following hours.

Set up: 18th

December 2014: 15:00-20:00

Take down: 20th

December 2014: 17:30-18:00

LUNCH

Reasonably priced lunch may be purchased in campus. For

food and beverage venues on campus, please kindly refer to

our attached Site Map.

TIPPING

Tipping is acceptable but not necessary in Hong Kong. In

restaurant, a 10% service charge is usually included unless

otherwise specified.

LOST AND FOUND

Please visit conference registration counter for lost and

found.

CLIMATE

With a sub-tropical climate, Hong Kong’s winter (December –

February) is cool and dry, with occasional cold fronts.

The average temperature for the winter season varies

between 12oC and 20

oC. It is often regarded by travelers as

the best time to travel to Hong Kong. Warm clothing is highly

recommended.

CURRENCY EXCHANGE

Hong Kong Dollar is acceptable in all stores. However,

Ranminbi is also accepted in most shops. One of the nearest

foreign currency exchange stores to conference venue is in

the paid concourse of MTR Shatin station.

HONG KONG AIRPORT AND TRANSPORT

As one of the world’s busiest passenger airports located on

Hong Kong Lantau Island, there are inevitable some of the

most convenient transportation systems available to provide

travelling to and from the airport.

The Mass Transit Railway (MTR) allows easy passage

throughout the New Territory, Hong Kong Island and Lantau

Island.

The Airport Express links this new airport with Kowloon and

Hong Kong Island, with trains departing every 10 minutes.

The one-way trip to MTR Hong Kong Station is around 24

minutes and HK$100.

The Chinese University of Hong Kong is located at the MTR

University station (East Rail Line). Taxi from airport to

University is around HK$300.

Courtyard by Marriott Shatin is located within walking

distance to MTR Shek Mun Station (Ma On Shan Line).

Alternatively, buses such as A41P can provide easy access to

the hotel from the airport, and vice versa. Shuttle bus is

available to deliver guests to the Chinese University of Hong

Kong in the morning. Other shuttle buses are also available

for regular destinations.

Hyatt Regency Shatin is located within walking distance to the

MTR University Station (East Rail Line). It is within walking

distance to the conference venue.

ELECTRICITY

Main supply voltage is 220 volts. Adapters are available in

most hotels.

CONFERENCE BANQUET On 19 Dec 2014, a conference banquet will be held at the

Star Seafood Floating Restaurant (沙田明星海鮮舫) at night.

Those who had fully registered and those who had paid the

banquet fees are eligible to attend.

RELEVANT WEBSITES PDT Conference: http://www.cuhk.edu.hk/scm/pdt2014/welcome.html CUHK Transportation: ttp://www.cuhk.edu.hk/english/university/visitors.html Hong Kong Airport: http://www.hongkongairport.com/eng/index.html Hong Kong Immigration Department: http://www.immd.gov.hk/en/home.html Mass Transit Railway: http://www.mtr.com.hk/

Welcome Message The 2014 International Conference on Photodynamic Therapy and Translational Medicine

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Welcome Message from the Organizer

On behalf of the Organizing Committee, I am delighted to welcome all speakers and participants to

the 2014 International Conference on Photodynamic Therapy and Translational Medicine at The

Chinese University of Hong Kong. The International Conference on Photodynamic Therapy in Hong

Kong was first held in 2001, which was well attended by international experts in photodynamic

therapy and diagnosis.

Recently, there have been rapid developments of photodynamic therapy and diagnosis from basic

research to clinical practice all over the world. Hong Kong is known as the gateway between the

East and the West. We are very grateful for the staunch support from the internationally renowned

experts in the fields of photodynamic therapy and diagnosis, and translational medicine from Hong

Kong, Mainland China, Asia and overseas speakers for the event. We aim to provide a platform for

the researchers, engineers, physicians, and graduate students worldwide to exchange your ideas

and strengthen regional and international collaborations.

I would also like to thank the members of the Organizing Committee and Local Organizing

Committee for their invaluable support and dedicated efforts, and the participants for attending

this unique conference.

We hope that all delegates will find this occasion a rewarding and memorable experience.

Albert LEUNG

Conference President

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Committees

iv

Conference Presidents

President: Albert LEUNG (Hong Kong, China)

Co-presidents: Stanley BROWN (UK)

Chuanshan XU (Hong Kong, China)

Honorary President: Dennis NG (Hong Kong, China)

Committees Organizing Committee

Zheng HUANG China Albert LEUNG Hong Kong, China Chuanshan XU Hong Kong, China

Scientific Committee (by alphabetical order)

Chairman Zheng HUANG China Members Mingdong HUANG China Albert LEUNG Hong Kong, China Quanhong LIU China Dennis NG Hong Kong, China Qian PENG Norway

Pan WANG Hong Kong, China Ricky WU Hong Kong, China Chuanshan XU Hong Kong, China Timothy ZHU USA

Local Organizing Committee (by alphabetical order) Chuanshan XU (Group Leader) Sze-Kei NG (Group Leader) Bella Wing-Yi CHENG Yue JIANG Heyu HUA Katy Ho-Kee KOON Dorian Wai-Keung LAU Siu-Kan LAW Winnie Wing-Sze LEE Winnie Wing-Yin LEE Kwan-Chi LEUNG Luva Wai-Ling LUI

Natalie Mei-Kam MOK Xin PANG Mandy Lap-Man TAM Lai-Lin TSANG Tammy TSE Pan WANG Xinna WANG Yin-Mei WONG Qicai XIAO Choice Lin-Choi YAU Angela Siu-Po YAU

Organizers and Sponsors The 2014 International Conference on Photodynamic Therapy and Translational Medicine

v

Organizers and Sponsors

Organizers

School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Centre for Photobiology and Photodynamic Therapy, University of Leeds, UK

MOE Key Laboratory of Optoelectronic Science & Technology for Medicine, Fujian Normal University

Supporting Organizations

International Photodynamic

Association European Platform for

Photodynamic Medicine Photodiagnosis and

Photodynamic Therapy

Sponsors

S. H. Ho College,

The Chinese University of Hong Kong Union Med. Limited

Ms. Po-Ching LEUNG Anonymous sponsor

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Site Map

vi

Site Map

Chung Chi College, The Chinese University of Hong Kong

Esther Lee Building Ground floor Lobby

Poster presentation Lecture Theatre 2

Oral presentation

Third Floor (ELB 308)

Workshop I

Possible Dining Venue *

Programme at a Glance The 2014 International Conference on Photodynamic Therapy and Translational Medicine

vii

Programme at a Glance

THURSDAY 18 DEC 2014 FRIDAY 19 DEC 2014 SATURDAY 20 DEC 2014

(8:45) Opening Ceremony

(8:45) Session I:

PDT and Translational Medicine I

(9:30) Plenary Lecture I

Stanley BROWN (UK)

(10:30) Coffee Break

(10:30) Coffee Break and Poster Session

(11:00) Plenary Lecture II (11:20) Session II:

PDT and Translational Medicine II

Zheng HUANG (China)

Chung-Ku RHEE (Korea)

(13:00) Lunch Break (12:30) Lunch Break

(14:30) Plenary Lecture III (14:00) Session: III

PDT Application and Technologies

Qian PENG (Norway)

Timothy ZHU (USA)

(15:20) Coffee Break

(16:30) Coffee break (16:00) Workshop II: Nanophotosensitizer and

PDT Application (17:00) Plenary Lecture IV

Yiru PENG (China) Zheng HUANG (China) Yiru PENG (China)

(15:00-20:00) Registration, Poster Posting

(18:30) Conference Banquet

(Star Seafood Floating Restaurant)

(17:30) Closing Ceremony

(18:00-21:30) Workshop I:

PDT Physics and Dosimetry

Timothy ZHU (USA)

Zheng HUANG (China)

* The organizer reserves the right to alter the topic/content/speaker of the conference programme without prior notice.

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Floor Plan

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Scientific Programme The 2014 International Conference on Photodynamic Therapy and Translational Medicine

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Scientific Programme

THURSDAY 18 DECEMBER 2014

Room 308, 3/F, Esther Lee Building, Chung Chi College, The Chinese University of Hong Kong

Start Time Topic and Speaker

15:00-20:00 Registration, Posting of Poster Papers

Workshop I: PDT Physics and Dosimetry

Timothy ZHU (USA)

18:00

Lecture I: Fundamental photophysics of PDT Lecture II: PDT dosimetry I- light dosimetry and transport of light in tissue Lecture III: PDT dosimetry II- Intracavitory and interstitial applications

19:45 Coffee Break

Zheng HUANG (China)

20:00

Lecture IV: PDT dosimetry III: topical application Lecture V: Understanding cutaneous phototoxicity

21:00 Q & A Sessions

21:30 End of Workshop I

See Course Introduction on p. 6

* The organizer reserves the right to alter the topic/content/speaker of the conference programme without prior notice.

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Scientific Programme

2

FRIDAY 19 DECEMBER 2014

Lecture Theater 2, Esther Lee Building, Chung Chi College, The Chinese University of Hong Kong


8:00 Registration

8:45

Opening ceremony

Welcoming Remarks

Dennis NG (Hong Kong, China)

Associate Vice-President, The Chinese University of Hong Kong

Albert LEUNG (Hong Kong, China)

Opening address Stanley BROWN (UK) Rongcheng LUO (China) (羅榮城,中國)

Qian PENG (Norway)

9:15 Group photos with Guests and Speakers

Plenary Lecture I

Chair: Albert LEUNG (Hong Kong China)

9:30

Stanley BROWN (UK) PDT, past, present and future

10:30 Coffee Break

Plenary Lecture II

Chair: Qian PENG (Norway)

11:00

Zheng HUANG (China) Vascular-targeted photodynamic therapy

12:00

Chung-Ku RHEE (Korea) Anti-bacterial and anti-biofilm PDT in otitis media

13:00 Lunch Break


3

FRIDAY 19 DECEMBER 2014 (CONT’D)


Plenary Lecture III

Chair: Stanley BROWN (UK)

14:30

Qian PENG (Norway) Photodetection and photodynamic therapy with porphyrin precursors

15:30

Timothy ZHU (USA) Singlet oxygen explicit dosimetry (SOED) for PDT

16:30 Coffee Break

Plenary Lecture IV

Chair: Zheng HUANG (China)

17:00

Yiru PENG (China) Block copolymer encapsulated poly (aryl benzyl ether) dendrimer silicon (IV) phthalocyanine for in vivo and in vitro photodynamic therapy of choroidal neovascularization

18:00 Close

18:30 Conference Banquet

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Scientific Programme

4

SATURDAY 20 DECEMBER 2014



Session I: PDT and Translational Medicine I

Chair: Timothy ZHU (USA)

8:45

9:05

9:25

9:45

10:05

Albert LEUNG (Hong Kong, China) Naturally occurring photosensitizers for photodynamic therapy Qing CHEN (Chongqing, China) AE-PDT induce cell death and inhibit metastasis in human breast cancer Jianmin HU (Xi’an, China) DVDMS is more potent than photofrin in photodynamic therapy in inhibiting breast cancer growth and metastasis in vitro and in vivo Chuan GU (Shanghai, China) The safety of photochemical tissue bonding for treating damaged cornea using limbal stem cells pre-cultured on human amniotic membrane Xinna WANG (Hong Kong, China) Investigation of photodynamically antitumor activity of hypericin upon LED light irradiation

10:30 Coffee Break and Poster Session

Session II: PDT and Translational Medicine II

Chair: Ricky WU (Hong Kong, China)

11:20

11:45

12:00

12:15

Ting-Pong FUNG (Hong Kong, China) The challenges in the development of clinical use of photodynamic therapy (PDT) in Hong Kong, a difficult and unusual path – Experience sharing of the pilot study in use of PDT for treatment of inoperable cholangiocarcinoma Haiping WANG (Xi’an, China) Studies on photo-sensitivity of a novel photosensitizer DVDMS and its anti-tumor efficacy Qicai XIAO (Hong Kong, China) An endogenous enzyme-NQO1-activated hypericin for photodynamic therapy Yue JIANG (Hong Kong, China) Preparation of hypocrellin B nanoparticles for photodynamic therapy

12:30 Lunch Break


5

SATURDAY 20 DECEMBER 2014 (CONT’D)


Session III: PDT Application and Technologies

Chairs: Chuanshan XU (Hong Kong, China), Katy KOON (Hong Kong, China)

14:00

14:20

14:40

15:00

Qingjuan TANG (Qingdao, China) Application of photodynamic inactivation technology in oyster processing Min YAO (Shanghai, China) Enzymatic activated PACT for methicillin-resistant staphylococcus aureus infection in vitro and in vivo Chuan WANG (Shanghai, China) Applications of 450-470nm visible light for killing P. aeruginosa, MRSA, and C. albicans Lin ZHOU (Nanjing, China) Silica nanoparticle-based drug delivery system of photosensitizers

15:20 Coffee Break

Workshop II: Nanophotosensitizer and PDT Application

Zheng HUANG (China)

16:00 Lecture VI: Fundamental - Photosensitizer and nanophotosensitizer

Yiru PENG (China)

Lecture VII: PDT Nanophotosensitizer I- Synthesis and photophysical property Lecture VIII: PDT Nanophotosensitizer II- In vitro and in vivo studies on nano-dendritic phthalocyanine photodynamic therapy

17:30 Close of Conference

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Workshops

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Workshops

Workshop I: PDT Physics and Dosimetry

Course Faculty

Timothy C. Zhu, PhD Professor of Radiation Oncology

Department of Radiation Oncology University of Pennsylvania School of Medicine (USA)

Zheng Huang, PhD Professor of Biomedical Photonics

MOE Key Laboratory of OptoElectronic Science & Technology for Medicine College of Photonics & Electronic Engineering

Fujian Normal University (China)

Course Introduction

Understanding the fundamental aspects of PDT physics and light dosimetry is critical in conducting the basic research and clinical application of PDT. This half-day (4 hours) workshop is carefully designed for both researchers and clinicians and will cover various aspects of PDT dosimetry by in-depth lectures and hands-on demonstration. The workshop will introduce the basics of photophysics and light dosemetry in PDT, provide an overview on the planning and monitoring of PDT dose, highlight the current state of art progress and challenges in PDT dosimetry, and discuss the importance of photophysical impact on biological effect and medical side effect of PDT. The workshop is suitable for both inexperienced and experienced PDT researchers and clinicians. All levels and all background attendants are welcome.

Workshop II: Nanophotosensitizer and PDT Application

Course Faculty

Yiru Peng, PhD Professor of Chemistry

College of Chemistry and Chemical Engineering, Fujian Province Key Laboratory of Polymer Materials, Fujian Normal University, Fuzhou, China

Zheng Huang, PhD

Course Introduction

The workshop will introduce the current progress and challenges of new photosensitizer and nanophotosensitizer. Using block copolymer encapsulated dendritic phthalocyanine photosensizers as an example, this course will provide some insight into the synthesis and potential applications of nanophotosensitizer with multimode potentials. In addition, in vitro and in vivo experimental studies on nano-dendritic phthalocyanine mediated photodynamic therapy of choroidal neovascularization will be discussed in detail.

List of Posters The 2014 International Conference on Photodynamic Therapy and Translational Medicine

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List of Posters No. Abstract Title Presenting Author

P01 Photodynamic therapy of human undifferentiated thyroid carcinoma AHN Jin-Chul

P02

FosPeg®-mediated photocytotoxicity suppressed cancer cell growth through down-regulation of p38MAPK protein

CHU Ellie Shihng-Meir

P03

Energy metabolism targeted drugs synergize with photodynamic therapy to potentiate breast cancer cell death

FENG Xiaolan

P04

Antibacterial efficacy of ultrasonic irrigation with sodium hypochlorite combined with photodynamic therapy in bovine root canals infected by Enterococcus faecalis

HUANG Xiaojing

P05

Inhibitory effect of F7-based photodynamic therapy on the growth of NCI-H460 human lung cancer cell xenograft in nude mice

JIANG Zhihuan

P06

Sonodynamic therapy with hypocrellin B on methicillin-resistant Staphylococcus aureus (MRSA)

LEUNG Albert

P07 Comparison of photobleaching characters for two photosensitizers LIU Hanqing

P08 Design and synthesis of amino acid-modified phthalocyanine LU Shan

P09 Photochemical activation cross-linking for stiffening cornea PENG Yinbo

P10

Comparison of the photodynamic activity of two cationic zinc phthalocyanines substituted with alanine

WANG Ao

P11 Sonodynamic action of hypocrellin B on Enterococcus faecalis WANG Xinna

P12

Comparison of the antitumor activity of nonionic, cationic and zwitter ionic phthalocyanine

WEI Shaohua

P13

Investigation of sonodynamically antibacterial chemotherapy of hyporellin B on biofilm-producing Staphylococcus epidermidis

XU Chuanshan

P14

Sonodynamic action of protoporphyrin IX against Staphylococcus aureus: Dark toxicity and bacterial growth inhibition

XU Chuanshan

P15

Fabrication of microscope compatible laser device for photodynamic therapy (PDT) in malignant brain tumor cases

YOSHIMITSU Kitaro

P16 In vitro anticancer activity of photosensitizer-encapsulated silica nanocage ZHOU Jiahong

P17 Photodynamic therapy by in situ nonlinear photon conversion PENG Xiao

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Keynote Speakers

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Keynote Speakers

Professor Stanley BROWN

Professor Emeritus, University of Leeds, UK

Professor Zheng HUANG

Center for Medical Photonics, Fujian Normal University, Fuzhou, China

School of Engineering and Applied Science, University of Colorado Denver, USA

Professor Qian PENG

Department of Pathology, Oslo University Hospital, University of Oslo, Oslo, Norway

Professor Yiru PENG

College of Chemistry and Chemical Engineering, Fujian Province Key Laboratory of Polymer Materials, Fujian Normal University, Fuzhou, China

Professor Chung Ku RHEE

Medical Laser Research Center and Department of Otolaryngology-HNS, Dankook University, Cheonan, Korea

Professor Timothy ZHU

Department of Radiation Oncology, Perelman Center for Advanced Medicine, University of Pennsylvania, USA

Plenary Lectures The 2014 International Conference on Photodynamic Therapy and Translational Medicine

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Abstracts – Plenary Lectures PDT, past, present and future

Stanley BROWN Professor Emeritus, University of Leeds, UK

Photodynamic therapy (PDT) involves the combined action of a photosensitiser, light and molecular oxygen

to kill cells in a targeted manner. All three of these components are required to achieve a PDT effect. The

technique has been developed to eliminate unwanted cells from the body, especially cancer cells and

microbial cells, but also unwanted vascular cells. Thus, PDT is being developed as an anti-cancer agent, as an

antimicrobial agent and as a treatment in ophthalmology. A cross-section of all of these applications was

discussed in details at the very successful PDT conference held in Hong Kong in December 2001. PDT has

been established since the late 1980s and, following large Phase III trials, several photosensitisers have now

been approved for PDT of cancer, including bladder cancer, head and neck cancer, lung cancer, oesophageal

cancer and skin cancers. However, there remains a need to improve these treatments (including

development of novel photosensitisers and new light sources) and the uptake of PDT into standard oncology

still needs to be optimised. On the other hand, PDT in ophthalmology (for age-related macular degeneration)

has been extremely successful and offered the first real hope for many thousands of patients suffering from

the wet form of this disease. In the antimicrobial field, a large amount of laboratory-based studies have

been carried out, but relatively few clinical studies and no large-scale applications of antimicrobial PDT have

yet been approved by regulators. This talk will briefly consider the history of PDT and how it has progressed

since the last Hong Kong conference in 2001, with an appraisal of what the future holds for PDT and related

technologies such as photochemical internalisation (PCI).

Vascular-targeted photodynamic therapy

Zheng HUANG1,2 1College of Photonic and Electronic Engineering, Fujian Normal University, Fuzhou, China

2 School of Engineering and Applied Science, University of Colorado, Denver, USA

Vascular-targeted photodynamic therapy (PDT) is of growing interest and probably by far the most

successful PDT application worldwide. It is characterized by a short drug-to-light interval (DLI) after the

completion of intravenous injection of photosensitizer. Under this unique approach, light irradiation takes

place while photosensitizers are still circulating in the vascular compartment. The interactions between light

and photosensitizers accumulated in the endothelial cells and bound to the vessel wall can cause vascular

effects through the low-density lipoprotein receptor-mediated endocytosis pathways and selective

endothelial cell damages, ultimately, lead to thrombosis and micro-vessel occlusion. Vascular-targeted PDT

has been used primarily for the management of several non-malignant diseases, such as the

neovascularization lesion (e.g. Visudyn® for wet age-related macular degeneration) and congenital capillary

malformations (e.g. HMME® for port wine stain birthmarks). Recently, vascular-targeted PDT has been

investigated for curative or palliative treatment of solid tumors (e.g. Tooka® soluble for prostate cancer) by

targeting the tumor vasculature. Although the photosensitizers used in vascular targeting mode might not

be selectively accumulated in cancerous cells during short DLI (typically 0-30 min) at all, vascular-targeted

PDT-mediated massive shutdown of pathological and normal vessels can deprive the supply of oxygen and

nutrients to tumor and subsequently achieve tumor ablation. This presentation will highlight the recent

development in vascular-targeted PDT.

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Plenary Lectures

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Antibacterial effects of PDT on bacteria of otitis media with effusion (OME)

Chung-Ku RHEE Medical Laser Research Center & Department of Otolaryngology-HNS, Dankook University, Cheonan-city, Korea

The aim of this study was to evaluate antibacterial effects of PDT on common bacteria causing otitis media with

effusion (OME). In vitro study was carried out using a hematoporphyrin derivative sensitizer (photogem) and 632

nm laser diode (LD) laser on S. pneumoniae, H. influenzae, and M. catarrhalis. One ml of each bacterial suspension

was incubated for 3 hours and various concentrations of photogem were administered into the suspension. The

suspensions were irradiated with 632 nm LD laser (15 J/cm2). The presence of colony forming units of the

bacteria was examined, cytometry of nucleic acid of bacteria was performed, and bacteria were observed by TEM.

The PDT was effective in killing all 3 kinds of bacteria. The flow cytometry showed lower intensity of bacterial

nucleic acid, and TEM showed damaged bacterial cell membrane and cytoplasmic structures. In vivo PDT study

was performed using gerbil. S. pneumoniae or H. influenza was injected into bullae. Photogem was injected into

bullae in 2 days when OME was developed and transcanal irradiation of 632 nm LD laser (90 J/cm2) was given.

Four days after PDT, middle ear and bullae were washed with DPBS and the washed DPBS was cultured. The

presence of bacterial colonies was examined. PDT was effective to kill S. pneumoniae in 87 % of the bullae with

OME while it was effective to eradicate H. influenzae in 50 % of the bullae with OME. The results of these studies

demonstrated that PDT may be effective to treat otitis media.

The effect of PDT on H. influenza biofilm in vitro and in vivo

Chung-Ku RHEE, So-Young CHANG, Phil-Sang CHUNG, Jin-Chul AHN Medical Laser Research Center & Department of Otolaryngology-HNS, Dankook University, Cheonan,-city, Korea

Biofilm formation has been demonstrated for many mucosal pathogens such as Haemophilus influenzae. The

presence of mucosal biofilms with chronic otitis media with effusion (COME) suggests that bacteria do not clear

by antibiotics. Objectives: To test the effect of photodynamic therapy (PDT) on H. influenzae induced biofilm in

vitro and in vivo. Methods: In vitro: Sixteen biofilms of H. influenzae were maintained on flow cell system culture

plates. The biofilms were divided into control, laser, photofrin, and PDT groups. For laser group, 7.2 J/cm2 (4 mW

x 30 min) of 632 nm LED was irradiated to the biofilms. For photofrin group, photofrins 5 and 25 μg/ml were

added to the media. For PDT group, photofrins 5 and 25 μg/ml were added to the media and LED 7.2 J/cm2 was

irradiated to the biofilms. Then the biofilms were cultured for 3 hours. Live/Dead (DAPI/PI) stain was performed

and biofilms were examined under confocal laser microscope for thickness and density of biofilms. In vivo:

Sixteen bullae of 8 gerbils were injected with 200 μl (107 CFU/ml) of H. influenza and formation of biofilms in the

bullae was obtained by 5 days. The bullae were divided into control, laser, photofrin, and PDT groups. The

control group received no treatment. For laser group, 120 J/cm2 (100 mW x 20 min) of 632 nm LD laser was

irradiated into the bullae by a fiber inserted directly into the bullae. For photofrin group, photofrin 40 μl (1

mg/ml) were injected into the bullae. For PDT group, similar to photofrin group, photofrin was injected into the

bullae and LD laser was irradiated into the bullae same way as in laser group. The mucosal tissues in bullae were

examined by H/E staining, and SEM. Results: In vitro: By PI staining 40 - 52 % reduction of biofilms in PDT group

was noted. The thickness of the biofilms were as followings; control group (124.3±11.7μm), laser group

(118.5±18.6 μm), photofrin 5 μg group (110 μm), photofrin 25 μg group (71 μm), PDT group with 5 μg photofrin

(110 μm), PDT group with 25 μg photofrin showed no growth of H. influenzae. In vivo: The control, laser, and

photofrin groups have shown well-formed biofilm. Two-third of the PDT group bullae have shown well resolved

biofilm while 1/3 of the bullae have shown incompletely resolved biofilms. Conclusion: The results of this study

demonstrated that PDT appears to be effective to treat experimental H. influenzae induced biofilms in vitro and in

vivo. Further trial in different dose combinations of photosensitizer and laser needs to be tried for better results

in PDT group. Clinical implication: PDT may be an alternative to antiobiotic treatment on otitis media with biofilm

formation.

Plenary Lectures The 2014 International Conference on Photodynamic Therapy and Translational Medicine

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Photodetection and photodynamic therapy with porphyrin precursors

Qian PENG Department of Pathology, Oslo University Hospital, University of Oslo, Oslo, Norway

Photodynamic Therapy (PDT) typically involves systemic administration of a lesion-localising photosensitiser

and its subsequent activation by visible light. This results primarily in a singlet oxygen-induced photodamage

to the lesion. Results from preclinical and clinical studies conducted worldwide over a 40-year period have

established PDT as a new clinical treatment modality of a number of malignant and non-malignant disorders.

PDT with chemically synthesized photosensitisers, however, has a major side-effect of skin phototoxicity,

limiting clinical PDT to a great extent. Considerable interest has been directed towards the development of a

new PDT regimen that relies on an endogenously synthesized photosensitiser. In the first step of the heme

biosynthetic pathway 5-aminolevulinic acid (ALA) is formed from glycine and succinyl CoA. The last step is

the incorporation of iron into protoporphyrin IX (PpIX, a potent photosensitiser) and takes place in the

mitochondria under the action of the rate-limiting enzyme, ferrochelatase. By adding exogenous ALA, the

naturally occurring PpIX may accumulate because of the limited capacity and/or low activity of

ferrochelatase. Porphobilinogen deaminase is another enzyme of the heme synthesis pathway (catalyzing

the formation of uroporphyrinogen from porphobilinogen). Its activity is higher in some tumours; while that

of ferrochelatase is lower, so that PpIX accumulates with a high degree of selectivity in these tumours. Such

selectivity has therefore been exploited for its application in photodetection (PD) and PDT of tumour.

Clinically, with porphyrin precursors PD of bladder cancer and glioma and PDT of actinic keratosis and

superficial basal cell carcinoma of the skin have already been approved by FDA and EU. This presentation will

update the clinical development of PD/PDT with porphyrin precursors, together with its possible mechanism

of action.

Singlet oxygen explicit dosimetry for PDT

Timothy ZHU Department of Radiation Oncology, University of Pennsylvania, USA

Objectives: Dosimetry of singlet oxygen (1O2) is of particular interest because it is the major cytotoxic agent

causing biological effects for type II photosensitizers during photodynamic therapy (PDT). We have

developed method to determine the reacted singlet oxygen threshold concentration (also called singlet

oxygen threshold dose) (1O2)sh for PDT based on explicit dosimetry of light and photosensitizer (PS) dose

during PDT. Methods: An in vivo RIF tumor mouse model is used to correlate the necrosis depth to the

calculation based on explicit PDT dosimetry of light fluence distribution, tissue optical properties, and

photosensitizer concentrations. Inputs to the model include 5 photosensitizer specific photochemical

parameters along with (1O2)sh. Photosensitizer specific model parameters are determined for several type II

photosensitizers (Photofrin, BPD) and compared with others (mTHPC) from the literature. Results: The in

vivo (1O2)sh (standard deviation) are approximately 0.56 (0.26) and 0.72 (0.13) mM (or 3.6 x 107 and 4.6 x 107

singlet oxygen per cell per 1/e fractional kill) for Photofrin and BPD, respectively, assuming that the fraction

of singlet oxygen generated that interacts with the cell is 1. This compared with the reported 9x 108 per cell

per 1/e fractional kill in an in vitro MLL cell model. Conclusion: The experimental results of threshold singlet

oxygen concentration in in vivo RIF tumor model for Photofrin and BPD are about 20-15 times smaller than

those observed in vitro. A SOED model is established to perform PDT explicit dosimetry during PDT.

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Plenary Lectures

12

Block copolymer encapsulated poly (aryl benzyl ether) dendrimer silicon (IV) phthalocyanine for

in vivo and in vitro photodynamic therapy of choroidal neovascularization

Yiru PENG1, Zheng HUANG2, Limin CHEN3, Guxing XU3 1College of Chemistry and Chemical Engineering, Fujian Province Key Laboratory of Polymer Materials, Fujian Normal

University, Fuzhou, China 2College of Photonic and Electronic Engineering, Fujian Normal University, Fuzhou, China

3Department of Ophthalmology, The Affiliated First Hospital of Fujian Medical University, Fuzhou, China

Objectives: The aim of the study is to demonstrate the photodynamic efficacy and safety of nanoparticles

loaded with poly (aryl benzyl ether) dendrimer Silicon (IV) phthalocyanine by blocking copolymer in vitro and

in vivo treatment of choroidal neovascularization as well as to discuss its therapeutic mechanism. Methods:

Uptake of nano-particles in HUVEC cells and RPE cells were quantified by fluorescence emission method. The

anti-proliferative effects were determined by CCK-8 assay. The changes of morphological of cells and

mitochondrial membrane potential (Δψm) changes of the HUVEC and RPE cells were investigated by

confocal microscope. The death mode of the cells following PDT and the intracellular levels of reactive

oxygen species (ROS) was studied by flow cytometry method. The impact of nano-particles on the vascular

endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) expression were examined

by real time fluorescent quantitative PCR. The feasibility of laser-induced choroidal neovascularization in BN

rat were evaluated by fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA), optical

coherence tomography (OCT), light microscope and histochemistry after photocoagulation. To evaluate the

effects of nano-particles on CNV in the BN rats, PDT with verteporfin was performed on another group.

Fundus examination, FFA, OCT, light microscope, histochemistry and electron microscope were used to

evaluate the effects of nano-particles on CNV in the BN rats. Results: The intracellular uptake amount of

nano-particles by HUVEC or RPE reached peak at 10h and 5h respectively. Nano-particles-based PDT

induced significant HUVEC cell death and relatively less RPE cell death in dose dependent manner. The

highest phototoxic were obtained by 20 μg/ml nano-particles with 10 J/cm2. The nucleolus underwent

apoptosis were observed. Both in HUVEC cells and RPE cells the mitochondrial membrane potentials of PDT

groups were distinctly decreased compared with the normal control group. The death mode of them cells

nano-particles was found to be apoptosis. Laser-induced choroidal neovascularization in BN rat was setup.

CNV was found 14 days after photocoagulation, reaching the peak on day 28. The success rate of CNV

induced by laser power was 84%. Disciform dye leakage appeared in the FFA and ICGA at the irradiated

spots. OCT, light microscope and immuhistochemistry were also taken for double check of CNV. Conclusion:

The kinetic change of the content of nanoparticles in HUVEC and RPE cells presented difference.

Nanoparticles-based PDT could significant kill HUVEC cell and RPE cell death in dose dependent manner.

Nanoparticles could induce apoptosis in HUVEC and RPE cells. Diode laser of 532 nm can successfully

induced CNV in BN rats. Twenty one to twenty eight days after photocoagulation may be the optimal time

to perform PDT. Nanoparticles with proper parameters have a potential effect to close CNV in BN rat. The

effect of PDT with verteporfin is similar to that of nano-particles -PDT.

PDT and Translational Medicine The 2014 International Conference on Photodynamic Therapy and Translational Medicine

13

Abstracts – PDT and Translational Medicine Naturally occurring photosensitizers for photodynamic therapy

Albert LEUNG1, Chuanshan XU 1,2 1 School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong 2 Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China

Photodynamic therapy (PDT) is a promising alternative to treat malignancies and infectious diseases.

Photosensitive drugs (also called photosensitizers), light source and molecular oxygen are three main

components affecting the clinical outcome of photodynamic therapy. The currently used photosensitizers in

clinical practice are mainly from porphyrin derivatives. Growing evidence has shown that many plant and

traditional Chinese herbal medicine contain photosensitive compounds. The findings from our lab showed

that these naturally occurring photosensitizers from traditional Chinese herbal medicine (for example,

curcumin, hypericin, hypocrellin and pheophorbide-a) had significantly photodynamic activity on cancer cells

and pathogenic bacteria. These indicated that traditional Chinese herbal medicine might be a rich source for

screening new and more efficient photosensitizers.

Acknowledgement: This work was supported by Innovation and Technology Fund of Shenzhen (CXZZ20120619150627260).

AE-PDT induces cell death and inhibits metastasis in human breast cancer

Qing CHEN, Dingqun BAI Department of Rehabilitation, the First Affiliated Hospital of Chongqing Medical University, Chongqing, China Objectives: To investigate the mechanism of photodynamic therapy (PDT) with aloe-emodin (AE) as a new

photosensitizer induce death and inhibit cell metastasis in human breast cancer in vitro. Methods: Cell viability

after photodynamic treatment with a series of AE concentration, light doses and times was measured by WST8

assay. Cell proliferation was detected by Edu assay and colony-forming unit assay. The changes of cell

morphology were observed by phase contrast microscope, H-33342 and transmission electron microscopy. The

rate of apoptosis and necrosis and mitochondrial membrane potential were detected by Annexin V-FITC/PI

double staining assay and JC-1 staining. The capacity of adhesion, migration and invasion were measured by

WST8, transwell assay and F-actin staining. Western blots detected the expression of related protein such as Bcl-

2, caspase-9, caspase-3, BIP, CHOP, MMP2, MMP9, VEGF and Nrf2. Results: Analysis of cell viability and

proliferation evidenced that there was a dramatical depression after photodynamic treatment with a series of AE

concentration and light dose showed. We observed changes from predominant apoptosis to predominant

necrosis depending on the light dose used (12 and 19.2 J/cm2 respectively) and expression of Bcl-2, caspase-9,

caspase-3, BIP and CHOP demonstrated that the mechanism of apoptosis was involved of mitochondria and

endoplasmic reticulum death pathway. The results of adhesion, migration and invasion in breast cells

demonstrated that AE-PDT significantly inhibited adhesion, migration and invasion of MCF-7 cells. Analysis of

changes in cytoskeleton component (F-actin) evidenced cytoskeleton disorganization after treatment with AE-PDT.

MMP2, MMP9, VEGF and Nrf2 demonstrated an early increase and a late decrease in expression patterns. This

pattern was related to oxidative stress. Conclusion: The present results indicated that PDT with aloe-emodin

effectively suppressed cancer development in MCF-7 cells, suggesting AE its potential as a new photosensitizer in

PDT has the potential to become mainstream of cancer treatment.

The 2014 International Conference on Photodynamic Therapy and Translational Medicine PDT and Translational Medicine

14

DVDMS is more potent than photofrin in photodynamic therapy in inhibiting breast cancer growth and

metastasis in vitro and in vivo

Jianmin HU1, Pan WANG1,2, Wenli XIONG1, Yichen LIU1, Quanhong LIU1, Xiaobing WANG1 1 College of Life Sciences, Shaanxi Normal University, Xi’an, China

2 School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong

Objectives: To investigate the proliferation and metastasis inhibition effect of DVDMS mediated PDT in a highly

metastatic 4T1 cell line and corresponding mouse xenograft models. Methods: The Multi-Volume

spectrophotometer system and Fluorescence spectrophotometer were used to investigate the spectral

characteristics of DVDMS. Production of reactive oxygen species (ROS) was assessed by flow cytometry. Cell

viability of different treatments was evaluated by MTT assay and colony formation assay. Scratch wound and

transwell assay was adopted to evaluate Cell motility. Furthermore, F-actin cytoskeleton and ultrastructural

changes in the cell were studied by laser scanning confocal microscopy and scanning electron microscopy

respectively. In addition, the in vivo antitumor effects of DVDMS against tumor cells were analysed using a mouse

xenograft model. The tumor cell structure after DVDMS-PDT treatment was observed by hematoxylin and eosin

(H&E) staining. The expression of PCNA was detected by immunohistochemistry. Besides, GPT and GOT levels

were determined by using GPT or GOT assay kit. Results: Light irradiation with DVDMS elicited potent

phototoxicity in vitro. The phototoxic effect was mediated by reactive oxygen species (ROS) and was reduced by

ROS scavengers. Besides, PDT combined with DVDMS effectively inhibited the migration of 4T1 cells. In an in vivo

mouse xenograft model, PDT with DVDMS markedly prolonged the survival of the tumor-bearing animals and

inhibited tumor growth and lung metastasis, consistent with in vitro findings. Interestingly, DVDMS showed higher

anti-tumor efficacy than Photofrin. In addition, the primary toxicological studies indicated that DVDMS is

relatively safe to administer. Conclusion: These results suggested that DVDMS is a promising photosensitizer that

deserves further development for the cancer treatments in photodynamic therapy.

The safety of photochemical tissue bonding for treating damaged corneal epithelium using limbal stem

cells pre-cultured on human amniotic membrane

Chuan GU, Yinbo PENG, Min YAO Department of Burn and Plastic Surgery, No. 3 People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China

Objectives: To determine the safety of photochemical tissue bonding (PTB) for treating damaged corneal

epithelium using limbal stem cells (LSCs) pre-cultured on human amniotic membrane (HAM). Methods: LSCs,

isolated from rabbit eyes and identified by cell markers, were labeled with BrdU prior to the cultivation on

de-epithelialized HAM to fabricate grafts. Rabbit LSCD models were created and randomly divided into

groups for transplantation of fabricated grafts using sutures or PTB (n=10). Possible phototoxicity of PTB to

LSCs was analyzed in vitro and in vivo. Restoration of corneal epithelium was evaluated at 28 days after

grafting. The BrdU labeled LSCs, as well as their phenotype, on restored corneal epithelium were tracked and

evaluated by immunefluorescent staining. Results: Our results showed that phototoxicity did not occur in

the LSCs cultured on HAM after PTB in vitro. Transplantation of grafts with PTB restored the damaged

cornea epithelium effectively and no significant influences on LSC characteristics were found in both sutured

and PTB groups. BrdU positive cells were tracked at 28 days post grafting suggesting that the restored

epithelium was derived from the in vitro fabricated HAM/LSC graft. In addition, the corneal opacity and

neovascularization in the PTB group were significantly lower than that in the sutured group confirming our

previous results. Conclusion: PTB is a safe and potential strategy for securing LSC/HAM grafts that produces

with better outcomes than sutured attachment.


15

Investigation of photodynamically antitumor activity of hypericin upon LED light irradiation

Xinna WANG1, Cindy YEUNG2, Albert LEUNG1, Chuanshan XU1

1 School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong 2 University of Toronto, Toronto, Ontario, Canada

Objectives: To investigate photodynamically antitumor activity of hypericin upon a novel LED light irradiation.

Methods: Hepatocellular carcinoma cells (HepG2 cells), the tumor cell model, were incubated with hypericin (2

μM) for 3 h at 37°C, and irradiated for various time (2, 4, 8, 16 and 32 s) by a novel LED light source with the

wavelength of 590 nm and a dose of 22 mW/cm2. The viability of HepG2 cells was measured using MTT assay and

trypan blue staining after photodynamic treatment. The changes of the cell nucleus were observed using a

confocal laser scan microscope (CLSM) and Hoechst 33342 staining. The collapse of mitochondrial membrane

potential was also measured using CLSM with Rhodamine123 staining. Results: Photocytotoxicity of hypericin on

HepG2 cells was light-dose dependent and more than 90% cells in combined treated group were killed after LED

light irradiation for 32 s. Condensed nuclei and apoptotic bodies were observed in the cells treated by LED-

irradiated hypericin. The notable collapse of mitochondrial membrane potential of HepG2 cells was observed

after photodynamic treatment. Conclusion: LED-irradiated hypericin significantly killed HepG2 cells in vitro.

Mitochondrial dysfunction might be an important event in the photodynamic action of hypericin.

The challenges in the development of clinical use of photodynamic therapy (PDT) in Hong Kong, a

difficult and unusual path – Experience sharing of the pilot study in use of PDT for treatment of

inoperable cholangiocarcinoma

Ting-Pong FUNG, Derek Tak-Lap TAM Department of Surgery, United Christian Hospital, Hong Kong

Background and Objectives: Photodynamic therapy (PDT) has been demonstrated to be beneficial to inoperable

cholangiocarcinoma patients, however, this modality of treatment is still not widely used. In the hospitals run by

the Hospital Authority (HA), Hong Kong, the application of PDT in treatment of inoperable cholangiocarcinoma is

novel. The connection and opinions of the development of clinical practice of PDT from the experts need to be

explored. It is indeed a complicated procedure requiring the involvement of machines, photosensitizers and

medical professionals. Therefore, the development initiation in HA is difficult. We would like to share our

experience and challenges confronted during the development of clinical practice of PDT in Hong Kong. Methods:

Our hospital has tried to apply the PDT treatment modality for patients since 2007. Various obstacles had been

encountered. (1) Funding for the treatment. (2) Supply of the laser machine and accessories. (3) Supply of the

photosensitizer. (4) Clinical Trial pathway. (5) Recruitment of patients. Results: Due to the uncertain supply of

the photosensentizer from the pharmaceutical companies, the project proposal has to be revised and reapproved

by the Ethical Committee of our hospital and Drug Office of the Department of Health, HKSAR from time to time,

resulting in taking 7 years to complete the preparation work for the clinical trial of PDT. Our first clinical trial was

finally carried out on 29/10/2014 and the patient is doing well. However, the supply of the photosensentizer

remains insecure which probably hinders the progress of the clinical trial. Conclusion: The clinical outcome from

our first PDT case for inoperable cholangiocarcinoma is quite promising and more cases have to be recruited to

demonstrate the efficacy. However, the experience in our hospital demonstrated that it is difficult to develop

PDT for clinical use in Hong Kong. The active involvement of the local university parties is no doubt an urgent

need to make the PDT clinical trial pathway easier in Hong Kong.

Acknowledgements: We would like to take this opportunity to thank Professor Huang, Professor Brown and Prof Moghissi for

their valuable time and supports.

The 2014 International Conference on Photodynamic Therapy and Translational Medicine PDT and Translational Medicine

16

Studies on photo- and sono-sensitivity of a novel photosensitizer DVDMS and its anti-tumor efficacy

Haiping WANG1, Jianmin HU1, Wenli XIONG1, Pan WANG1,2, Quanhong LIU1, Xiaobing WANG1 1

College of Life Sciences, Shaanxi Normal University, Xi’an, China. 2 School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong

Sinoporphyrin sodium, also called DVDMS, was a photosensitizer which is separated from Photofrin. In this study,

the physical and chemical properties and photo- and sono- sensitivity are analyzed both in vitro and in vivo. The

spectral of DVDMS showed DVDMS was mainly in the monometric form in buffer solution and the pH and ionic

strength of solution had an effect on DVDMS spectral. DVDMS showed higher auto-fluorescence intensity and

singlet oxygen production efficiency as compared with other photosensitizers in both cancer cells and normal cells.

In addition, the primary safety analysis results indicated that the hepatic damage was not significantly observed in

photosensitizer-treatment groups. The photo-sensitivity analysis demonstrated that the maximal uptake of

DVDMS occurred within 3 hours, with a mitochondrial sub-cellular localization. MTT assay and colony formation

assay demstrated that DVDMS could be effectively activated by light and the phototoxicity was much higher than

that of Photofrin under the same conditions. Besides, PDT combined with DVDMS effectively inhibited the

migration of 4T1 cells. In an in vivo mouse xenograft model, PDT with DVDMS markedly prolonged the survival of

the tumor-bearing animals and inhibited tumor growth and lung metastasis, consistent with in vitro findings.

Interestingly, DVDMS showed higher anti-tumor efficacy than Photofrin. The sono-sensitivity of DVDMS are also

confirmed both in vitro and in vivo. Compared with Hp, DVDMS mediated SDT showed stronger cytotoxicity on

Eca-109 cells. Serious mitochondrial damage, Cyt C release, apoptosis rate enhancement and apoptosis protein

activation were detected after SDT treatment. The three-dimensional optical imaging system shows that DVDMS

has a preferential localization in tumors, but low accumulation in most of the normal tissues. A significant

synergistic effect of ultrasound combined with DVDMS was obtained when the load power was 4 W and DVDMS

dose was above 2 mg/kg. At the fourteenth day after DVDMS-SDT, the tumor volume inhibition ratio was 56.27%.

In addition, the tumor weight inhibition ratio after the synergistic treatment was 55.37% which was obviously

stronger than the ultrasound treatment alone (23.85%) and DVDMS alone (23.15%). Moreover, no metastasis

was happened to the tumors in the SDT-treated mice as compared with control group. All of these results clearly

demonstrated that DVDMS could be activated by light and ultrasound. The potential mechanism research found

that DVDMS-mediated PDT (DVDMS-PDT) induced DNA damage and mitochondrial membrane potential loss.

Moreover, there was abundant intracellular ROS found after the treatment with PDT or SDT, and the cytotoxicity

induced by PDT and SDT was effectively remitted by ROS scavenger. The generated ROS destroy mitochondria

and cause mitochondrial dependent apoptosis. Herein, DVDMS is a promising photosensitizer and sonosensitizer

that deserves further development for the cancer treatments in both photodynamic therapy and sonodynamic

therapy. What is more, DVDMS could be used as a sensitizer in sono-photodynamic therapy due to its sono- and

photo-sensitivity.

An endogenous enzyme-NQO1-activated hypericin for photodynamic therapy

Qicai XIAO, Albert LEUNG, Chuanshan XU School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong

Objectives: To improve the phototherapeutic efficacy by enhancing the reactive oxygen species generation and

the tumor selectivity. Methods: An endogenous enzyme-responsive photosensitizer was designed and

synthesized by conjugating a quinone based enzyme substrate to a natural photosensitizer. And the

photodynamic activity of the prepared molecule was evaluated via investigating its photophysical and

photochemical properties. Results: An endogenous enzyme-responsive photosensitizer was prepared and

characterized, the fluorescence was quenched by photo-induced electron transfer, and the water solubility was

greatly improved. Conclusion: Conjugation a quinone-based enzyme substrate to hypericin has a great influence

on the fluorescence, the photosensitizing property and the water solubility. The effects of its photodynamic

activity are being investigated.


17

Preparation of Hypocrellin B nanoparticles for photodynamic therapy

Yue JIANG, Albert LEUNG, Chuanshan XU School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong

Objectives: Hypocrellin B (HB), a naturally occurring photosensitizer (PS) from traditional Chinese herb, has a

very potent photodynamic antitumor effect. However, it is highly hydrophobic and can produce aggregates

in the physiological solution, which consequently reduces photodynamic efficacy. Apoferritin is a potential

nanovehicle in which PS can be packaged to improve the water solubility as well as avoid unfavorable

biodistribution of free PS. The objective of this work is to prepare HB nanoparticles to achieve a better

therapeutic outcome. Methods: HB-loaded apoferritin nanoparticles (HB-AFT NPs) were prepared by taking

advantage of the reversible unfolding-refolding character of apoferritin in different pH environments. High-

performance liquid chromatography (HPLC) was used to determine HB content and encapsulation efficiency.

The obtained HB-AFT NPs were also characterized by transmission electron microscopy (TEM), dynamic light

scattering (DLS), zeta potential (ζ) and UV/Vis spectrum. MTT assay was performed to investigate the in vitro

photodynamic therapy on breast cancer cell line MDA-MB-231. Results: Results from HPLC, DLS and TEM

indicate that HB molecules were successfully encapsulated within protein cages with about 85%

encapsulation efficiency. Upon such encapsulation, these HB-AFT NPs became water-soluble and kept the

original spectroscopic characters of HB and apoferritin. Interestingly, the phototoxicity to MDA-MB-231 cells

of the HB encapsulated apoferritin nanocages was markedly improved as compared to free HB at specific

light intensity. Conclusion: HB-AFT NPs can successfully increase the solubility of HB and significantly

improve photodynamic therapy of HB on MDA-MB-231 cells. The detailed mechanisms will be explored in

our further investigation.

The 2014 International Conference on Photodynamic Therapy and Translational Medicine PDT Application and Technologies

18

Abstracts – PDT Application and Technologies Application of photodynamic inactivation technology in oyster processing

Juan WU1, Binbin CAO1, Tao ZUO1, Changhu XUE1, Albert LEUNG2, Chuanshan XU2, Qingjuan TANG1 1 College of Food Science and Engineering, Ocean University of China, Qingdao, China


Objectives: The elimination of pathogenic microorganisms in shellfish is an area of increasing concern to

consumers. The effective microbial destruction is imperative in ensuring safety and extending shelf-life of

oyster. However, the traditional sterilization technologies have disadvantages to some extents. Therefore,

it makes an urgent need to seek new non thermal sterilization technologies for reducing the microbial load.

Photodynamic therapy (PDT) is an alternative way to eradicate pathogenic microorganisms. This study aimed

to investigate the application of photodynamic inactivation technology in oyster processing. Methods:

Photodynamic inactivation technology was applied in oyster processing with the treatment of curcumin.

Total bacteria and several epidemic pathogenic microorganisms in oysters such as Norovirus (NoV),

Escherichia coli (E. coli) and Vibrio parahaemolyticus were investigated both in vitro and in vivo. The

mechanism of inactivation was investigated using microbiological method. Results: Results showed that the

activities of microorganisms were substantially inhibited after photodynamic treatment of curcumin in a

dose-dependent manner. Notable damages to the nucleic acid and protein molecule were found in these

microorganisms. Conclusion: Photodynamic inactivation technology has a sufficient level of antimicrobial

activity in oyster processing. This technology is, for the most part, less energy-intensive and therefore more

cost-efficient and environmentally friendly than conventional processing.

Enzymatic activated PACT for methicillin-resistant Staphylococcus aureus infection in vitro and in

vivo

Xiujun FU1, Yunqing ZHU2, Min YAO1 1 Department of Burn and Plastic Surgery, No. 3 People's Hospital, Shanghai Jiao Tong University, Shanghai, China

2 School of Materials Science and Engineering, Tongji University, Shanghai, China

Objectives: To investigate the selective inhibition of methicillin-resistant Staphylococcus aureus (MRSA) by a

modified photosensitizer (LAEtNBS) in vitro and the efficacy of MRSA infection treatment by photodynamic

therapy (PDT) with LAEtNBS in vivo. Methods: Two bacteria strains MSSA and MRSA were used to

determine the efficacy and selectivity of PDT with various concentrations of LAEtNBS at a fluence of 15 J/cm2

with 640 ± 10 nm LED light. Human skin fibroblasts and keratinocytes were also applied to test the

phototoxicity of LAEtNBS. Further, mouse skin wound infected with 108 CFU of MRSA were treated by PDT

with LAEtNBS, EtNBS-COOH, or PBS control at a fluence of 90 J/cm2, and efficacy of bacterial inactivation as

well as wound healing were evaluated. Results: LAEtNBS was shown to have very similar absorption and

emission wavelength with EtNBS-COOH, but tremendously supressed fluorescence quantum yield and less

potency in singlet oxygen production. LAEtNBS was demonstrated to selectively inactivate MRSA in vitro,

and exert less cytotoxicity to host tissues than EtNBS-COOH. Both LAEtNBS and EtNBS-COOH were shown to

be able to reduce MRSA infection in mouse skin wound and enhance tissue repair, although LAEtNBS did not

show significant improvement than EtNBS-COOH. Conclusion: Modification of the photosensitizer by taking

advantage of enzymes expressed by specific bacteria might be ideal for targeting bacterial infection therapy

with less harm to host tissue and bacterial balance.

PDT Application and Technologies The 2014 International Conference on Photodynamic Therapy and Translational Medicine

19

Applications of 450–470 nm visible light for killing P. aeruginosa, MRSA, and C. albicans

Chuan WANG, Min YAO Department of Burn and Plastic Surgery, No. 3 People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

Objectives: To investigate the killing effects of 450–470 nm visible light (blue light) on P. aeruginosa,

methicillin-resistant Staphylococcus aureus (MRSA) and C. albicans. Methods: The planktonic P. aeruginosa,

MRSA, and C. albicans were irradiated with 12, 60, 120, and 240 J/cm2 of blue lights, respectively. The killing

effects were evaluated using tablet coated plate method and flow cytometry after irradiation. Blue light

with energy density of 240 J/cm2 was used to treat P. aeruginosa, MRSA, and C. albicans that inside biofilm.

The survival of cells and biofilm structure were assessed by confocal laser scanning microscope and scanning

electron microscope. A mouse skin wound model infected with C. albicans was created; the infected

wounds were then treated with 240 J/cm2 of blue light. The killing effect was determined at day 1, day 2 and

day 3 after irradiation and the wound-healing rate was detected at every another day up to 14 days after the

treatment. Results: The effects of blue light for killing planktonic P. aeruginosa, MRSA, and C. albicans

increased with higher energy densities, which started from 60 J/cm2 up to 240 J/cm2, compared with blank. P.

aeruginosa, MRSA, and C. albicans in biofilm were destroyed mostly and the biofilm structures became

sparse and incompact after treated with 240 J/cm2 of 450–470 nm visible light. In addition, the infection

with C. aibicans in animal skin wounds was controlled and wound-healing rate was accelerated by blue light

treatment. Conclusion: 450–470 nm visible light shows highly efficient killing of planktonic and biofilm P.

aeruginosa, MRSA, and C. albicans in vitro and it can be applied as an alternative therapy for C. albicans

infections in vivo.

Silica nanoparticle-based drug delivery system of photosensitizers Lin ZHOU College of Chemistry and Materials Science, Nanjing Normal University, Nanjing, China

Objectives: Photodynamic therapy (PDT) has attracted great attention in cancer treatment. However, many

photosensitizers (PSs) are hydrophobic. Therefore, for delivery of these PSs, special formulations are

required to make their aqueous dispersion, often using nanoparticle-based delivery systems. Methods:

Silica nanoparticle has been receiving great attentions because of its water dispersion ability, high stability

and compatibility with biological systems. Our researches indicated that many PSs can be encapsulated

inside silica nanoparticles to improve their water dispersion ability and PDT activity. Results: The result PSs

encapsulated silica-based nanoparticles are monodisperse and stable in aqueous solution. Comparative

studies with free PS and entrapped one have demonstrated that the encapsulation effect on the embedded

PSs significantly enhances their efficacy of singlet oxygen generation and, thereby, the in vitro photodynamic

efficacy. Furthermore, these silica nanoparticle-based drug delivery system of PSs can be modified by heavy

atoms, upconversion nanocrystals or graphene quantum dots to enhance PDT activity and prolong the

wavelength of sensitive light to near infrared or multi-color imaging. Conclusion: All these properties of

silica nanoparticle make it especially promising to be used as a drug delivery system for PSs in the field of

PDT.

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Poster Session

20

Abstracts – Poster Session Photodynamic therapy of human undifferentiated thyroid carcinoma

Jin-Chul AHN1,2,3, Raktim BISWAS1,2,3, Yun-Hee RHEE3, Arindam MONDAL3, Phil-Sang CHUNG3, Chung-Ku RHEE3, Jeong-Hwan MOON3 1 Department of Biomedical Science, Dankook University, Cheonan, South Korea 2

Biomedical Translational Research Institute, Dankook University, Cheonan, South Korea 3

Beckman Laser Institute Korea, Dankook University, Cheonan, South Korea

Thyroid cancer is the most common endocrine and ninth most common cancer overall. Among all of the

thyroid cancers, anaplastic thyroid cancer (ATC) is considered as one of the most aggressive malignancies

with very poor prognosis. Patients suffering from ATC can survive only a few months after the diagnosis

because this type of undifferentiated thyroid cancer is very aggressive in nature. Conventional treatments

like thyroidectomy and chemotherapy are not capable of complete recovery of the patients. Thus, several

alternative methods along with conventional methods have been introduced to treat anaplastic thyroid

cancer. Photodynamic therapy (PDT) has been studied against human anaplastic thyroid cancer for last few

years. In our present study, various thyroid cancer cells were treated with PDT. The efficacy of PDT has also

been studied in tumor xenograft model as well as orthotopic thyroid cancer model. Results showed

promising efficacy of PDT in anaplastic thyroid cancer treatment. Several apoptosis related proteins were

expressed in PDT treated cells. Expressions of several growth factor related proteins also were modulated in

PDT treated group. For in vitro experiments, several parameters like cytotoxicity assay and confocal

microscopic study revealed that the number of apoptotic cell was higher in treated groups as compared to

the control group. For in vivo xenograft study, the tumor volume was reduced in a greater extent in the PDT

treated group than the no treatment group. Expressions of several proteins related to cell proliferation and

growth were modulated by PDT treatment. Efficacy of PDT was also studied in orthotopic thyroid cancer

model. Anaplastic thyroid cancer cells were injected into the thyroid region of nude mice. Mice were

treated with PDT irradiating the tumor region with laser. Reduction in tumor volume can be observed in PDT

treated group. There were significant differences in the mean tumor volume between the treated and

control group after PDT. Therefore, we can conclude that PDT with various photosensitizers can be used as

an effective treatment method to control the aggressive anaplastic thyroid cancer as well as other types of

cancer.

Poster Session The 2014 International Conference on Photodynamic Therapy and Translational Medicine

21

FosPeg®-mediated photocytotoxicity suppressed cancer cell growth through down-regulation of

p38MAPK protein

Ricky Wing-Kei WU1,2, Ellie Shihng-Meir CHU2, Zheng HUANG3, Chuanshan XU4, Christine Miu-Ngan Li YOW1

1 Medical Laboratory Science, Department of Health Technology & Informatics, Hong Kong Polytechnic University, Hong Kong 2 Department of Medical Science, Tung Wah College, Hong Kong

3 Department of Medicine, National Jewish Health, Denver, USA


Background and Objectives: FosPeg® is a new formulation of mTHPC contained in pegylated liposomes with

optimized distribution properties. We previously reported that FosPeg® mediated Photodynamic Therapy

(PDT) was effective to EBV positive human nasopharyngeal carcinoma cell line c666-1 and EBV negative

human nasopharyngeal carcinoma cell line HK1. However, the FosPeg® PDT efficacy depends on its tumour

localization, light absorption spectrum and the environmental oxygen content. Therefore, studying the

localization and the biological targets of FosPeg® become very important to understand the PDT-mediated

mechanism of cellular destruction in tumours. In this in vitro study, the localization of FosPeg®, potential

efficacy of FosPeg® PDT and its modulation on p38 MAPK proteins on both cell lines were investigated.

Methods: The localization of FosPeg® on c666-1 cells and HK1 cells were determined by fluorescence

microscopy. Effects of FosPeg® PDT on p38 and phosphorylated p38MAPK protein were examined by

Western blot analysis. Results: FosPeg® was mainly localized in the mitochondria but not in lysosome, Golgi

Complex and Endoplasmic Reticulum in both cell lines. Down-regulation of p38 and phosphor-p38MAPK

proteins were observed in both cell lines after FosPeg® PDT in a dose dependent manner. Results were

correlated with the FosPeg® PDT induced cell cycle progression and DNA content change as reported

previously. Conclusion: FosPeg®-mediated PDT exerted antitumor effect on c666-1 cells and HK1 cells

through down-regulation of p38MAPK signal pathways. Understanding the mechanism of FosPeg® mediated

PDT may help to develop improved strategies for the treatment of NPC.

Acknowledgement: We would like to thank Prof. Kwok-Wai LO from the Department of Anatomical and Cellular Pathology, CUHK for

providing NPC/c666-1 and NPC/HK1 cells and Biolitec for providing FosPeg®.


22

Energy metabolism targeted drugs synergize with photodynamic therapy to potentiate breast

cancer cell death

Xiaolan FENG1, Yi ZHANG1, Quanhong LIU1, Xiaobing WANG1, Pan WANG1, 2

1 College of Life Sciences, Shaanxi Normal University, Xi’an, China


Objectives: To elucidate the general potentiation of photodynamic therapy (PDT) efficacy by 2-DG and 3-

bromopyruvate (3-BP) and the underlying mechanism of the process in human breast cancer MDA-MB-231

cells. Methods: The via-count assay was adopted to examine cytotoxicity of different treatments. Changes

on apoptosis, mitochondrial membrane potential and intracellular reactive oxygen species (ROS) production

were analysed using a flow cytometer. The following apoptosis-related responses such as Bax translocation

and the expression of Bax/Bcl-2, caspase-3 and mitogen-activated protein kinase (MAPK) were detected and

analysed by immunofluorescence and Western blotting assays. Results: Under optimal conditions, the

combined treatments (2-DG+PDT and 3-BP+PDT) significantly aggravated PDT-induced MDA-MB-231 cell

death when compared with either monotherapy. Synergistic potentiation on cell apoptosis, MMP loss and

ROS generation were observed. In addition, the combined treatments caused the increase of cell cytotoxicity

and cell apoptosis, MMP loss, and ROS formation were all differently inhibited by N-acetylcysteine (NAC).

Conclusion: The results indicate that 2-DG and 3-bromopyruvate (3-BP) have a general potentiation of PDT

efficacy in human breast cancer cells, and the combination treatments induced mitochondrial and caspase-

dependent cell apoptosis, in which ROS may play an important role.

Antibacterial efficacy of ultrasonic irrigation with sodium hypochlorite combined with

photodynamic therapy in bovine root canals infected by Enterococcus faecalis

Suli XIAO, Dianfu MA, Yanhuang WANG, Shan JIANG, Xiaojing HUANG School and Hospital of Stomatology, Fujian Medical University, Fuzhou,, China

Objectives: To evaluate the antibacterial efficacy of ultrasonic irrigation (UI) using different concentrations

of sodium hypochlorite (NaOCl) combined with photodynamic therapy (PDT) on root canals of bovine teeth

infected with Enterococcus faecalis (E. faecalis) in vitro. Methods: One hundred and twenty bovine teeth

infected with E. faecalis were randomly divided into 4 main groups as follows for decontamination: (1)

Irrigated with saline; (2) Methylene blue (MB)-mediated PDT; (3) UI with NaOCl (0.5%, 1.0%, 2.0%, 2.5% or

5.25%) alone; (4) UI with NaOCl (0.5%, 1.0%, 2.0%, 2.5% or 5.25%) combined with MB-mediated PDT. Colony

forming unit counting was performed respectively. The data were analysed with one-way ANOVA and LSD

post-hoc tests (P < 0.05). Results: The antibacterial efficacy of all experimental groups was higher than that

of saline group (P < 0.05). UI with NaOCl (5.25%), or combination of UI with NaOCl (2.0%, 2.5% or 5.25%)

and MB-mediated PDT eliminated E. faecalis more effectively than the other treatment groups (P < 0.05.

Additionally, UI with NaOCl (2.0% or 2.5%) combined with MB-mediated PDT were more efficient in killing E.

faecalis UI with NaOCl (2.0% or 2.5%) alone (P <0.05). Conclusion: UI with NaOCl (2.0% or 2.5%) combined

with MB-mediated PDT are effective and potentially an alternative of UI with NaOCl (5.25%) to avoid its side

effects in root canal disinfection.


23

Inhibitory effect of F7-based photodynamic therapy on the growth of NCI-H460 human lung

cancer cell xenograft in nude mice

Zhihuan JIANG1, Aiping WANG2, Wentao WU3 1 New Drug Safety Evaluation Center, Chinese Academy of Medical Sciences and Peking Union Medical College

2 Institue of Materia Medica, Chinese Academy of Medical Science, Beijing, China

3 Tianjin Ocelean Pharmaceutical Science andTechnology Co., Ltd, Tianjin, China

Objectives: To observe the inhibitory effect of F7-based photodynamic therapy (PDT) on the growth of NCI-H460 human lung cancer cell xenograft in nude mice. Methods: NCI-H460 tumor mass was inoculated subcutaneously into the right flank of nude mice (NU/NU). After 7 days, the 42 tumor-bearing nude mice were randomly divided into the following groups: (1) non-treatment control, (2) solvent control, (3) positive control with 20 mg/kg Photofrin®, (4) – (6) treatment groups with F7 at doses of 0.3, 0.6 and 1.2 mg/kg. Groups (2) – (6) were treated with irradiation condition 76 J/cm2 (127.3 mW/cm2, 600 s) 24 h after photosensitizer injection. During the recovery time, tumor length and width were measured every two days until the average tumor volume exceeded 2.0 cm3 at which time all the mice were euthanized. Results: Compared with non-treatment control group, the inhibitory effect on the average tumor weights in positive control and F7-PDT (0.6, 1.2 mg/kg) groups were significant (P < 0.05). The tumor weight inhibition rates of (4) – (6) groups with F7-PDT were 23.89%, 66.94% and 92.34% (positive control, 86.89%). T/C values of groups (4) – (6) were 45.79%, 30.30% and 10.88% (positive control, 9.68%). Conclusion: Treatment with F7-PDT (0.6, 1.2 mg/kg) results in a significant inhibition of H460 tumor growth.

Sonodynamic therapy with hypocrellin B on methicillin-resistant Staphylococcus aureus (MRSA)

Albert LEUNG1, Xinna WANG1, Margaret IP2, Chuanshan XU1 1 School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong 2 Department of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong

Objectives: The present study aims to investigate sonodynamic therapy with hypocrllin B on methicillin-

resistant Staphylococcus aureus (MRSA). Methods: The bacterial growth was determined using the colony

counting method. The synthesis of nucleic acid in MRSA and intracellular reactive oxygen species (ROS) were

also measured after sonodynamic treatment of hypocrellin B. Results: Survival of MRSA significantly

decreased as the intracellular ROS level significantly increased after sonodynamic treatment. However, no

significant change in the synthesis of nucleic acid was found after the treatment. Conclusion: Sonodynamic

action of hypocrellin B significantly increased intracellular ROS level and inhibited the survival of MRSA.

Acknowledgement: This work was supported by Health and Medical Research Fund (1100502) and the general research fund (GRF)

grant from Hong Kong research grant committee (RGC) (476912). We express our sincere thanks to Ms. Irene Ang and Ms. Miuling

Chin for their helpful assistance.


24

Comparison of photobleaching characters for two photosensitizers

Hanqing LIU1, Hongtao JIN1,2, Aiping WANG 1,2 1 Institute of Materia.Chinese Acdemy of Medical Sciences & Peking Union Medical College, Beijing, China

2 Beijing Union-Genius Pharmaceutical Technology Development Co., Ltd.

Objectives: The aim of this study is to compare the photobleaching characteristics of Sinoporphyrin sodium

(DVDMS-2) with a-(8-QLO)PcZn)[a-(8-quinolinoxy)zinc phthalocyanine (F7). Methods: Absorption curve of

DVDMS-2 and F7 were measured using Micro-Ultraviolet Spectrophotometer. The concentrations of

DVDMS-2 and F7 were set as 0.1 – 80 µg/mL, using the laser output power of 1500 mW, spot diameter of 8

cm and irradiation time of 3 – 180 min. The concentrations of the photosensitizers were detected using

Colorimetric Method. The photobleaching of DVDMS-2 and F7 were calculated and compared. Results: (1)

The absorbance and the concentrations of the DVDMS-2 and F7 showed good linear relationship at

maximum absorption wavelength within the scope of 0.1 – 80 μg/mL. The photobleaching rate is positively

related to the concentration of DVDMS-2 and F7 when other conditions were fixed. Longer duration of the

light irradiation increased the photobleaching volume but reduced the photobleaching per unit time. The

photobleaching rate of F7 was apparently higher than DVDMS-2 under the same conditions. No significant

photobleaching were observed under the conditions of pharmacodynamics studies (DVDMS: 20 μg/mL, 1500

mW × 3 min; F7: 12 μg/mL, 1500 mW ×3 min). The photobleaching rates of F7 in different solutions were:

normal saline solution < cell suspension < no-serum medium solution < medium solution with 10% fetal calf

serum (FBS) and the photobleaching rates of DVDMS-2 in different solutions were: cell suspension < normal

saline solution < no-serum medium solution < medium solution with 10% FBS. Conclusion: Both DVDMS-2

and F7 were not bleached under the conditions of pharmacodynamic studies, which ensured the efficacy of

photodynamic therapy (PDT). The photobleaching rate of F7 was higher than that of DVDMS-2, means F7

needs less avoiding-light time than DVDMS-2 after PDT.

Design and synthesis of amino acid-modified phthalocyanine

Shan LU, Ao WANG, Li GUI, Lin ZHOU, Shaohua WEI College of Chemistry and Materials Science, Nanjing Normal University, Nanjing, China

Objectives: Photodynamic therapy (PDT) is an effective treatment of cancer. The efficiency of PDT crucially

depends on the nature of the photosensitizers (PSs). Phthalocyanines (Pcs) are promising second-generation

PSs for PDT because of their strong absorption in tissue-penetrating red light and high efficiency of

generating singlet oxygen (1O2). However, the poor solubility of the Pcs in physiological fluids has

significantly hindered their clinical applications. Therefore, we wish to disclose new amino acid-incorporated

phthalocyanine sensitizer, which displays good water solubility, low tendency of aggregation, low toxicity,

favourable biocompatibility. Methods: Three novel amino acid-modified zinc(II) phthalocyanines were

synthesized. They were characterized by using UV-Vis, IR and 1H NMR. Microscopic and nuclear chromatin

fluorescence images of control cells and light triggered Pcs treated HeLa cells were obtained. Results:

Drastic changes in the morphology were obtained when the HeLa cells were treated with drugs overnight.

The cells shrunk and emerged many membrane blebs. The chromatin fluorescence of the control cells

stained dimly and occupied the majority of the cell nucleus. In contrast, the cells treated with amino acid-

modified zinc(II) phthalocyanines and irradiation showed obvious morphological changes, such as chromatin

shrinkage, chromatin condensation and fragmentation. Conclusion: The amino acid-modified zinc(II)

phthalocyanines are promising anticancer photosensitizers.


25

Photochemical activation cross-linking for stiffening cornea

Yinbo PENG1, Ti WANG2, Jingyin ZHU3 1 Department of Burn and Plastic Surgery, 3rd People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China 2 Department of Ophthalmology, The 85th Hospital of PLA, Shanghai, China 3 Department of Ophthalmology, Huadong Hospital Affiliated to Fudan University, Shanghai, China

Objectives: To explore the effect of photochemical activation induced corneal cross-linking, Rose Bengal (RB) and

532 nm green light irradiation (RB-PCL) were utilized and evaluated on porcine corneal biomechanical rigidity and

the biochemical resistance against collagenase digestion. Methods: A protocol with a wavelength of 532 nm and

illumination intensity of 0.4 W/cm2 for 250 s to deliver a dose of 100 J/cm2 was chosen. We demonstrated the

diffusion depth of RB into porcine cornea by using confocal microscopy. After photochemical cross-linking, an

increase in tensile strength and Young’s modulus in porcine corneas was observed. A corneal buttons with a

diameter of 1 cm was treated by collagenase type 2 to observe the digestion time. Results: Using confocal

microscopy, we demonstrated that the diffusion depth of RB into porcine cornea was approximately 150 μm and

mostly localized in anterior stroma 25 min followed by RB application. After photochemical cross-linking, an

increase in tensile strength (by average 200%) and Young’s modulus (by average 200%) in porcine corneas was

observed. The corneal buttons treated by RB-PCL showed doubling of collagenase digestion time from 10.8 ± 3.1

days in the blank group to 19.7 ± 6.2 days in the RB-PCL group, indicating increased resistance to enzymatic

digestion. Conclusion: Collagen cross-linking by RB-PCL increased both of the biomechanical stiffness and the

biochemical resistance against collagenase digestion in porcine corneas, which allow stabilization and

solidification of the cornea. The advantages and disadvantages of RB-PCL versus UVA/riboflavin cross-linking

technique (UV-CXL) are fully explored. Due to the nature of minimal penetration of RB into corneal stroma, the

RB-PCL method could potentially be used in patients with corneal thickness less than 400 mm where UV-CXL is

limited.

Comparison of the photodynamic activities between two cationic zinc phthalocyanines

substituted with alanine

Ao WANG, Shaohua WEI School of Chemistry and Materials Science, Nanjing Normal University, Nanjing, China

Objectives: Through the comparison of the photodynamic activities between two cationic zinc phthalocyanines

(quaternized 2(3), 9(10), 16(17), 23(24)-tetra-((4-(N-(2-amino) propanamide) amino) phenoxyl) phthalocyaninato-

zinc (ZnPc1) and quaternized 2(3), 9(10), 16(17), 23(24)-tetra-(4-(N-(2-amino) propanamide) methylamino)

phenoxy) phthalocyaninato-zinc (ZnPc2)), to provide a new attempt for the preparation of water soluble and low

aggregation zinc phthalocyanines, which can be used for photodynamic therapy. Methods: The photophysical

and photochemical properties (such as aggregation degree, photostability and fluorescence properties) of the two

cationic phthalocyanines were investigated by UV-Vis and fluorescence spectrum. Furthermore, the efficacy of

singlet oxygen generation was measured by using disodium salt of 9, 10-anthracenedipropionicacid (ADPA) as the

probe. Through the addition of base and acid, the effects of Ph on the properties of the two phthalocyanines were

studied. Results: The pH had different effects on the properties of ZnPc1 and ZnPc2. ZnPc1, which exhibited a

tautomeric transformation between amide and imidohydrine when it was catalysed by base, showed a lower

aggregation and better photodynamic activity and anticancer activity comparing to ZnPc2. Conclusion: The design

and properties regulation of phthalocyanines should be based on their structures, while a minor change in the

structure would be useful in some cases.


26

Comparison of the antitumor activity of nonionic, cationic and zwitter ionic phthalocyanine

Li GUI, Shaohua WEI College of Chemistry and Materials Science, Nanjing Normal University, Nanjing, China

Objectives: Photodynamic therapy (PDT) is a promising method for the treatment of various oncological

diseases. Phthalocyanines (Pcs), as the photosensitizers (PSs), have lots of advantages in PDT. Yet, very few

studies have been reported the relationship between PDT activity and the ionic nature of Pcs. Methods:

There novel Pcs, including tetra-substituted octa-cationic zinc(II) phthalocyanine(ZnPc1), zwitter ionic zinc(II)

phthalocyanine (ZnPc2) substituted with 2-((4-methoxyphenyl)dimethyl ammonio)ethanesulfonate and one

non-ionic zinc(II) phthalocyanine (ZnPc3) substituted with methoxy-N, N-dimethylaniline moieties, were

successfully synthesized and their fluorescence intensity, intracellular uptake, reactive oxygen levels and in

vitro photodynamic activities of the three Pcs were compared. Results: The result indicates that ZnPc1

analogue exhibits strongest singlet oxygen generation ability among three ZnPcs. Upon illumination, ZnPc1

has higher fluorescence intensity, intracellular uptake, reactive oxygen levels and in vitro photodynamic

activities towards HeLa cells as comparing with the other two zinc phthalocyanines (ZnPc2, ZnPc3).

Conclusion: The ionic nature of Pcs could affect their photodynamic activity and the cationic Pcs has superior

PDT activity than non-ionic and zwitter ionic Pcs in our experiment.

Sonodynamic action of hypocrellin B on Enterococcus faecalis

Xinna WANG1, Margaret IP2, Albert LEUNG1, Chuanshan XU1

1 School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong 2 Department of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong

Objectives: The present study aims to investigate sonodynamic action of hypocrllin B on Enterococcus

faecalis. Methods: The uptake of hypocrellin B in Enterococcus faecalis was measured using fluorescence

intensity analysis. The bacterial growth was determined using the colony counting method and bacterial

membrane integrity was analyzed by a flow cytometry with propidium iodide staining. Chromosomal DNA of

bacteria was observed using a pulsed-field gel electrophoresis (PFGE). Results: The uptake of hypocrellin B

in Enterocuccus faecalis reached a high peak at the 20 min incubation. Survival of Enterocuccus faecalis

significantly reduced and membrane integrity was remarkably damaged after sonodynamic treatment of

hypocrellin B. However, no significant change in chromosomal DNA was found after sonodynamic treatment.

Conclusion: Sonodynamic action of hypocrellin B increased bacterial membrane integrity, and inhibited the

survival of Enterocuccus faecalis. The detailed mechanisms needed to be clarified in our future investigation.

Acknowledgement: This work was supported by the general research fund (GRF) grant from Hong Kong research grant committee

(RGC) (476912) and Direct Grant of the Chinese University of Hong Kong (4053026). We express our sincere thanks to Ms. Irene Ang

and Ms. Miuling Chin for their helpful assistance.


27

Investigation of sonodynamically antibacterial chemotherapy of hypocrellin B on biofilm-

producing Staphylococcus epidermidis

Chuanshan XU1, Xinna WANG1, Margaret IP2, Albert LEUNG1

1 School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong 2 Department of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong

Objectives: The present study aims to investigate sonodynamically antibacterial chemotherapy of

hypocrellin B on biofilm-producing Staphylococcus epidermidis. Methods: The time course of hypocrellin B

in biofilm-producing Staphylococcus epidermidis was measured using fluorescence intensity analysis. After

sonodynamic treatment of hypocrellin B, the bacterial growth was determined using the colony counting

method. Bacterial membrane integrity was analysed by a flow cytometry with propidium iodide staining.

Intracellular reactive oxygen species (ROS) level was also detected using a flow cytometry with DCFH-DA

staining. Results: The uptake of hypocrellin B in biofilm-producing Staphylococcus epidermidis reached a

plateau at the 50-min-incubation. Hypocrellin B-mediated sonodynamic action significantly induced survival

reduction of Staphylococcus epidermidis. Bacterial membrane integrity was remarkably damaged and

intracellular ROS level was markedly increased after sonodynamic treatment of hypocrellin B. Conclusion:

Sonodynamic action of hypocrellin B increased intracellular ROS level, damaged bacterial membrane

integrity, and significantly killed biofilm-producing Staphylococcus epidermidis.

Acknowledgement: This work was supported by the general research fund (GRF) grant from Hong Kong research grant committee

(RGC) (476912) and Direct Grant of the Chinese University of Hong Kong (4053026). We express our sincere thanks to Ms. Irene Ang

and Ms. Miuling Chin for their helpful assistance.

Sonodynamic action of protoporphyrin IX against Staphylococcus aureus: Dark toxicity and

bacterial growth inhibition

Chuanshan XU1,2*, Jinghui DONG1, Albert LEUNG1, Margaret IP3 1 School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

2 Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China

3 Department of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong

Objectives: To investigate the dark toxicity and bacterial growth inhibition of protoporphyrin IX (PpIX) on

Staphylococcus aureus. Methods: Staphylococcus aureus was exposed to sonodynamic treatment of PpIX.

Dark toxicity and bacterial growth inhibition of PpIX were measured using colony forming units assay.

Results: PpIX had no remarkable toxicity to Staphylococcus aureus in the dark. The combined treatment of

PpIX and ultrasound sonication had significant inhibition on the growth of Staphylococcus aureus. However,

PpIX treatment alone and ultrasound sonication alone had no significant effect on the growth of

Staphylococcus aureus in this study. Conclusions: Sonodynamic action of PpIX had significant inhibition on

the growth of Staphylococcus aureus

Acknowledgements: This project was supported by the general research fund (GRF) grant from Hong Kong research grant committee

(RGC) (476912), Direct Grant of the Chinese University of Hong Kong (4053026) and Innovation and Technology Fund of Shenzhen

(CXZZ20120619150627260). We express our sincere thanks to Ms. Irene Ang and Ms. Miuling Chin for their helpful assistance.


28

Fabrication of microscope compatible laser device for photodynamic therapy (PDT) in malignant brain tumor cases Kitaro YOSHIMITSU1, Yoshihiro MURAGAKI2, Ken MASAMUNE 1, Yoshihiro FUKUTOMI1, Takashi MARUYAMA2, Hiroshi ISEKI1,3 1 Faculty of Advanced Techno-Surgery, Tokyo Women’s Medical University, Tokyo, Japan

2 Department of Neurosurgery, Faculty of Advanced Techno-Surgery, Tokyo Women’s Medical University, Tokyo, Japan

3 Faculty of Science and Engineering, Waseda University, Tokyo, Japan

Objectives: Clinical trial for photodynamic therapy (PDT) in malignant brain tumor had performed since 2007

at Tokyo Woman’s Medical University and was approved in September 2013. PDT is one of the cutting edge

therapies by using 664 nm laser equipment with talaporfin sodium. Currently, the remarkable therapy is only

available in the selected hospitals that are equipped with the microscopes produced by Mitaka Kohki Co., Ltd.

Since the laser unit must be mounted on the microscope with means, the application of this unique

therapeutic performance is limited. To compensate this problem for widespread use, we report a novel

microscope compatible laser equipment for PDT. Methods: The first prototype was fabricated by

prototyping with 3D printer. The therapeutic laser probe was inserted into a 2.5 mm diameter and a 200 mm

stainless steel tube and the tip was guided manually by adjusting the miniature 3D stage. The tip was

surrounded by three liner laser guides which are located consisting triangular pyramid. Results: The

targeting point of the laser probe is geometrically positioned 25 mm below. The main body is attached at the

end effector of the flexible multi-joint arm which is fixed on the rail by the surgical bed. Additionally, the

whole unit is covered with a sterilized plastic sheet and a disposable plastic part, so that the laser probe was

enabled to be kept its cleanliness after sterilization. The minor improvement for the hardware is shortening

the distance between each liner laser guides for considering narrow operative field and another distance

between the miniature 3D stage and laser unit considering its stableness for clinical use. Major hardware

improvement would be required based on the evaluated results. Conclusion: This paper reported the first

prototyping fabrication of the therapeutic laser device for PDT. The verification of the targeting accuracy by

this newly developed device will be performed as in future.

In vitro anticancer activity of photosensitizer-encapsulated silica nanocage processing

Lin ZHOU, Jiahong ZHOU Analysis and Testing Centre, Nanjing Normal University, Nanjing China

Objectives: Photodynamic therapy (PDT) is a light triggered cancer treatment method. However, many

commonly used photosensitizers (PSs) are hydrophobic, which makes their stable formulation for systemic

administration highly challenging. To overcome this difficulty, effective strategies have to be evolved to

enable a stable dispersion of these drugs into aqueous systems. Methods: Our group has prepared and

characterized a serial of silica nanocage-based drug delivery system of PSs using different methods, including

so-gel method and micro-emulsion method. Results: It was indicated that these silica nanocage-based drug

delivery system of PSs could greatly improve the water dispersion of hydrophobic PSs. Furthermore, the

light stability, reactive oxygen species generation ability and in vitro anticancer activity of PSs were greatly

improved after being encapsulated inside silica nanocages. Conclusion: All the properties of these

nanocages make this drug delivery system of PSs possible to be applied in the field of PDT.


29

Photodynamic therapy by in situ nonlinear photon conversion

Xiao PENG, Danying LIN, Junle QU Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of

Optoelectronic Engineering, Shenzhen University, Shenzhen, China

We present the development of a multidimensional optical method which integrates in a single experimental

setup emerging optical technologies. The proposed platform allows us to simultaneously collect

multidimensional parameters of a sample, thereby opening up new pathways for molecular detection in live

cells and deeper understanding of molecular mechanism of cellular processes, disease manifestation and

progression. Using this platform, we recently developed a new form of photodynamic therapy (PDT) in

which nonlinear optical interactions of near-infrared laser radiation with a biological medium in situ produce

light that falls within the absorption band of the photosensitizer. By modelling and experiment, we

demonstrate activation of a known photosensitizer by in situ nonlinear-optical up-conversion of the near-

infrared laser radiation using Second Harmonic Generation (SHG) in collagen and Four-Wave Mixing (FWM),

including Coherent anti-Stokes Raman Scattering (CARS), produced by cellular biomolecules. Introduction of

CARS/FWM to PDT in vitro increases the efficiency two-times compared to two-photon PDT alone, while SHG

provides a five-fold increase.

Key words: multi-dimensional; multimodal nonlinear optical microscopy; CARS; FLIM; FWM; PDT

The 2014 International Conference on Photodynamic Therapy and Translational Medicine List of Participants

30

List of Participants

No. Family Name Given Name Department / Institution Country Email

1 AHN Jin-Chul Department of Biomedical Science, College of Medicine, Dankook University

Korea [email protected]

2 BAI Dingqun Department of Rehabilitation, The First Affiliated Hospital of Chongqing Medical University

China [email protected]

3 BROWN Stanley Centre for Photobiology and Photodynamic Therapy, University of Leeds

UK [email protected]

4 CHEN Qing Department of Rehabilitation, The First Affiliated Hospital of Chongqing Medical University


5 CHU Ellie Shihng-Meir

Department of Medical Science, Tung Wah College Hong Kong, China

[email protected]

6 DENG Xiaofeng Department of General Surgery, Xiangya 2nd Hospital, Central South University


7 FENG Xiaolan College of Life Sciences, Shaanxi Normal University China [email protected]

8 FUNG Ting-Pong Department of Surgery, United Christian Hospital Hong Kong, China

[email protected]

9 GU Chuan Department of Burn and Plastic Surgery, NO.3 People's Hospital, Shanghai Jiao Tong University


10 HE Hui Soochow University China [email protected]

11 HU Jianmin College of Life Sciences, Shaanxi Normal University China [email protected]

12 HUA Heyu School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Hong Kong, China

[email protected]

13 HUANG Mingdong Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences


14 HUANG Xiaojing School and Hospital of Stomatology, Fujian Medical University


15 HUANG Zheng

College of Photonic and Electronic Engineering, Fujian Normal University College of Engineering and Applied Sciences, University of Colorado

China USA

[email protected]

16 ISEKI Hiroshi Faculty of Science and Engineerying, Waseda University

Japan [email protected]

17 JI Meng China Medical (Tianjin) Group Ltd. Co. China [email protected]

18 JIANG Shan School and Hospital of Stomatology, Fujian Medical University


19 JIANG Yue School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Hong Kong, China

[email protected]

20 JIANG Zhihuan New Drug Safety Evaluation Center in Chinese Academy of Medical Sciences & Peking Union Medical College


21 JIN Liwei Marketing Department of Shanghai Fudan-Zhangjiang Bio-Pharmaceutical Co.,Ltd.


22 KANG Ling College of Public Health, Xinjiang Medical University China [email protected]

23 KOON Ho Kee School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Hong Kong, China

[email protected]

24 LAW Siu-kan School of Chinese Medicine, Faculty of Medicine, The Chinese University of Kong Kong

Hong Kong, China

[email protected]

25 LEUNG Albert Wing-nang

School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Hong Kong, China

[email protected]

26 LIN Jiwang Department of General Surgery, Xiangya 2nd Hospital, Central South University


27 LIU Hanqing Institute of Materia, Chinese Academy of Medical Sciences & Peking Union Medical College


28 LIU Quanhong College of Life Sciences, Shaanxi Normal University China [email protected]

29 LIU Xinlong China Medical (Tianjin) ST Group Co.,LTD China [email protected]

30 LU Shan College of Chemistry and Materials Science, Nanjing Normal University


List of Participants The 2014 International Conference on Photodynamic Therapy and Translational Medicine

31

List of Participants (CONT’D)

No. Family Name Given Name Department / Institution Country Email

31 LUO Rongcheng Traditional Chinese Medicine-Integrated Hospital, Southern Medical University


32 LV Yuanyuan Marketing Department of Shanghai Fudan-Zhangjiang Bio-Pharmaceutical Co.,Ltd.


33 LV zhiping School of Traditional Chinese Medicine, Southern Medical University


34 NAKAMURA Naoto Panasonic Healthcare Co., Ltd. Japan [email protected]

35 PANG Xin School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Hong Kong, China

[email protected]

36 PENG Qian Department of Pathology, Oslo University Hospital, University of Oslo

Norway [email protected]

37 PENG Xiao

Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University


38 PENG Yinbo Department of Burn and Plastic Surgery, NO.3 People's Hospital, Shanghai Jiao Tong University


39 PENG Yiru College of Chemistry and Chemical Engineering, Fujian Normal University


40 RHEE Chungku Medical Laser Research Center, Department of Otolaryngology-HNS, Dankook University

Korea [email protected]

41 TANG Qingjuan College of Food Science and Engineering, Ocean University of China


42 WANG Ao School of Chemistry and Materials Science, Nanjing Normal University


43 WANG Binzhe China Medical (Tianjin) ST Group Co.,LTD China [email protected]

44 WANG Chuan Department of Burn and Plastic Surgery, NO.3 People's Hospital, Shanghai Jiao Tong University


45 WANG Haiping College of Life Sciences, Shaanxi Normal University China [email protected]

46 WANG Pan School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong


47 WANG Xiaobing College of Life Sciences, Shaanxi Normal University China [email protected]

48 WANG Xinna School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Hong Kong, China

[email protected]

49 WEI Shaohua College of Chemistry and Materials Science, Nanjing Normal University


50 WU Ricky Wing-Kei Department of Medical Science, Tung Wah College, Hong Kong

Hong Kong, China

[email protected]

51 XIAO Qicai School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Hong Kong, China

[email protected]

52 XU Chuanshan School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong

Hong Kong, China

[email protected]

53 YAO Min Department of Burn and Plastic Surgery, NO.3 People's Hospital, Shanghai Jiao Tong University


54 YOSHIMITSU Kitaro Faculty of Advanced Techno-Surgery, Tokyo Women's Medical University

Japan [email protected]

55 ZHOU Jiahong Analysis and Testing Centre, Nanjing Normal university


56 ZHOU Lin College of Chemistry and Materials Science, Nanjing Normal University


57 ZHU Timothy Department of Radiation Oncology, Perelman Center for Advanced Medicine, University of Pennsylvania

USA [email protected]

mailto:[email protected]

mailto:[email protected]

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Index of Authors

32

Index of Authors

Ａ

AHN, Jin-Chul ........................................................ 10, 20

Ｂ

BAI, Dingqun ............................................................... 13

BISWAS, Raktim .......................................................... 20

BROWN, Stanley ........................................................... 9

Ｃ

CAO, Binbin ................................................................. 18

CHANG, So-Young ....................................................... 10

CHEN, Limin ................................................................ 12

CHEN, Qing ................................................................. 13

CHU, Ellie Shihng-Meir ................................................ 21

CHUNG, Phil-Sang ................................................. 10, 20

Ｄ

DONG, Jinghui ............................................................. 27

Ｆ

FENG, Xiaolan.............................................................. 22

FU, Xiujun .................................................................... 18

FUKUTOMI, Yoshihiro ................................................. 28

FUNG, Ting-Pong ........................................................ 15

Ｇ

GU, Chuan ................................................................... 14

GUI, Li .................................................................... 24, 26

Ｈ

HU, Jianmin .......................................................... 14, 16

HUANG, Xiaojing ......................................................... 22

HUANG, Zheng ................................................. 9, 12, 21

Ｉ

IP, Margaret .................................................... 23, 26, 27

ISEKI, Hiroshi ............................................................... 28

Ｊ

JIANG, Shan ................................................................. 22

JIANG, Yue .................................................................. 17

JIANG, Zhihuan ............................................................ 23

JIN, Hongtao ................................................................ 24

Ｌ

LEUNG, Albert .................... 13, 15, 16, 17, 18, 23, 26, 27

LIN, Danying ................................................................ 29

LIU, Hanqing ................................................................ 24

LIU, Quanhong ................................................ 14, 16, 22

LIU, Yichen................................................................... 14

LU, Shan ...................................................................... 24

Ｍ

MA, Dianfu .................................................................. 22

MARUYAMA, Takashi .................................................. 28

MASAMUNE, Ken ........................................................ 28

MONDAL, Arindam ...................................................... 20

MOON, Jeong- Hwan .................................................. 20

MURAGAKI, Yoshihiro ................................................. 28

Ｐ

PENG, Qian ................................................................. 11

PENG, Xiao .................................................................. 29

PENG, Yinbo .......................................................... 14, 25

PENG, Yiru ................................................................... 12

Ｑ

QU, Junle ..................................................................... 29

Ｒ

RHEE, Chung-Ku .................................................... 10, 20

RHEE, Yun-Hee ............................................................ 20

Ｔ

TAM, Derek Tak-Lap .................................................... 15

TANG, Qingjuan .......................................................... 18

Index of Authors The 2014 International Conference on Photodynamic Therapy and Translational Medicine

33

Index of Authors (CONT'D)

Ｗ

WANG, Aiping ....................................................... 23, 24

WANG, Ao ............................................................. 24, 25

WANG, Chuan ............................................................. 19

WANG, Haiping .......................................................... 16

WANG, Pan ..................................................... 14, 16, 22

WANG, Ti .................................................................... 25

WANG, Xiaobing ............................................. 14, 16, 22

WANG, Xinna .............................................15, 23, 26, 27

WANG, Yanhuang ....................................................... 22

WEI, Shaohua .................................................. 24, 25, 26

WU, Juan ..................................................................... 18

WU, Ricky Wing-Kei .................................................... 21

WU, Wentao ............................................................... 23

Ｘ

XIAO, Qicai .................................................................. 16

XIAO, Suli .................................................................... 22

XIONG, Wenli ........................................................ 14, 16

XU, Chuanshan ............. 13, 15, 16, 17, 18, 21, 23, 26, 27

XU, Guxing .................................................................. 12

XUE, Changhu ............................................................. 18

Ｙ

YAO, Min ......................................................... 14, 18, 19

YEUNG, Cindy .............................................................. 15

YOSHIMITSU, Kitaro .................................................... 28

YOW, Christine Miu-Ngan Li ....................................... 21

Ｚ

ZHANG, Yi .................................................................... 22

ZHOU, Jiahong ............................................................ 28

ZHOU, Lin ........................................................ 19, 24, 28

ZHU, Jingyin ................................................................ 25

ZHU, Timothy ............................................................. 11

ZHU, Yunqing .............................................................. 18

ZUO, Tao ..................................................................... 18

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Notes

34

Notes The 2014 International Conference on Photodynamic Therapy and Translational Medicine

35

The 2014 International Conference on Photodynamic Therapy and Translational Medicine Notes

36

The 2014 International Conference on Photodynamic Therapy and Translational Medicine

december 18 to 20, 2014 the chinese university of hong kong · table of contents the 2014...

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