research article determination of serum lost goodwill...

Research ArticleDetermination of Serum Lost Goodwill Target Proteome inPatients with Severe Traumatic Brain Injury

Hongming Ji1 Changchen Hu1 Gangli Zhang12 Jinrui Ren1 Yihu Tan3 Wenxiao Sun3

Junwen Wang4 Jun Li4 Hongchao Liu3 Ruifan Xie2 Zhipeng Hao3 and Dongsheng Guo2

1Department of Neurosurgery Shanxi Provincial Peoplersquos Hospital Taiyuan 030001 China2Department of Neurosurgery Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China3Department of Neurosurgery Liyuan Hospital Tongji Medical College Huazhong University of Science and TechnologyWuhan 430077 China4Department of Neurosurgery Wuhan Central Hospital Tongji Medical College Huazhong University of Science and TechnologyWuhan 430014 China

Correspondence should be addressed to Gangli Zhang zhanggangli1973163com

Received 8 March 2015 Revised 26 July 2015 Accepted 28 July 2015

Academic Editor Aijun Wang

Copyright copy 2015 Hongming Ji et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

This study investigates the biokinetics of LGT proteome a potential biomarker of severe TBI in serum of severe TBI patients TheLGT proteome presents in the serum of severe TBI patients The abundance diversity of LGT proteome is closely associated withpathologic condition of TBI patients Serum LGT proteome may be used as a promising marker for evaluating severity of severeTBI

1 Introduction

Traumatic brain injury (craniocerebral injury TBI) is themost common disease with high morbidity and mortalityin neurosurgery especially to the youngsters and a lot ofresearchers have focused on this fact [1 2] Severe TBI repre-sents a major cause of neurological morbidity and mortalitythroughout the world In the past decades several challengeshave been faced in the conduct of clinical research in TBIincluding inclusion of a broad heterogeneity of injuriesdifficulties with standardization and consistency of complexmedical management and lack of sophisticated measuresto sufficiently detect the differences in outcomes Recentdevelopments in definition of potential genetic and biologicalmarkers in TBI are also aiding in the subcategorization ofpatients into finer diagnostic and prognostic groups [3]

Serum protein biomarkers have long held promise in thestudy of TBI They are considered to facilitate the diagnosismonitoring progress prognosis and providing pertinentmolecular information about ongoing pathological changesfor designing evidence-based therapeutic interventions [4]

Serum protein biomarkers are of special importance in TBIbecause they are typically associated with military operationswith limited access to imaging and other diagnostic toolsof hospitals [5 6] So there is great significance to look forthe related protein markers to the TBI patientsrsquo pathogeneticcondition and prognosis [2 7] Pei [8ndash10] reported a kindof proteome called lost goodwill target (LGT) proteomein the serum of the tumor patients which can reflect thedifferent phase of tumor development or aggravation whichwas also related to the patientsrsquo pathogenetic condition andmortality Ren et al [11] reported that the LGT proteome wasproduced under the pathologic condition of TBI patientsand the abundance of LGT proteome is closely associatedwith pathogenetic condition and prognosis of TBI patientsthe LGT proteome may play an important role in predictingpathogenetic condition and prognosis of TBI patients

Based on the findings above LGT can also be found insevere TBI patientsrsquo serum and it may be the early warningsign for disease aggravation or even death In order to inves-tigate the clinical significance of LGT proteome the studyproceededwith surface enhanced laser desorptionionization

Hindawi Publishing CorporationBioMed Research InternationalVolume 2015 Article ID 183821 7 pageshttpdxdoiorg1011552015183821

2 BioMed Research International

time-of-flight mass spectrometry (SELDI-TOF-MS) [12 13]technology which can detect the exact LGT concentration inthe serum of TBI severe patients

2 Materials and Methods

21 General Data 96 TBI cases treated in our departmentbetween March 2006 and July 2009 were analyzed retrospec-tively There were 77 males and 19 females with age range of16ndash83 years (mean age 317plusmn263) interval time from injuryto clinical (h) 05sim115 h average 71 plusmn 22 Glasgow ComaScale (GCS) score 44 cases with 3sim5 points 52 cases with 6sim8points and 32 cases receiving emergency operation traumacause 46 cases by traffic accident 22 cases by injury due tofall or mine disaster 19 cases by strike injury and 6 caseswith other causes major trauma type 47 cases with contu-sion and laceration of brain and (or) subduralintracerebralhematoma 14 cases with epidural hematoma and 35 caseswith diffuse axonal injury and brain stem injury All caseswere diagnosed as craniocerebral trauma by cranial CTscanning after the admission 69 survival cases carried onprognostic evaluation with Glasgow outcome score (GOS)6 months later The exclusion criteria were as follows beingunder 16 years old death in 24 hours from admission severeinfection or combined injury malignant tumor or autoim-mune disease with damage of important organs especiallyliver and kidney (multiple organ failure) and transfusionof blood in operation Meanwhile 35 healthy cases werechosen as control group This project has been reviewed bythe Medical Ethics Committee of Shanxi Provincial PeoplersquosHospital At the same time signed informed consent ofresearch projects of every patient with patients themselvesor their guardians was obtained The factors such as generalcondition age and sex were comparable (in research groupand the normal controls) between the two groups (119875 gt 005)

22 SELDI-TOF-MS Detection

221 Main Instruments and Reagents Themain instrumentsand reagents included protein fingerprint meter WCX2chips PBS-IIc chip reading machine and Ciphergen Pro-teinChip 32 analysis software urea sodium acetate [NaAc(100mmolL NaAc pH 40)] HPLC water and SPA

222 Serum Sample Processing 5mL sera were taken inthe morning when patients were hollow and were putin VACVTTE vacuum quantitative mining container andplaced at room temperature for 60min (including the timewhen samples were out of the body) and then the serumwas separated from blood at 3000 g times 10min and preservedat minus80∘C serum samples were thawed and put in ice bath(10000 g times 2min) take 5 120583L serum sample and add 10 120583L urea(9mmolL) The sample underwent oscillation for 30minfollowed by the addition of 180 120583L buffer (100mmolL NaAcpH = 40) at 4∘C and was centrifuged at 10000 g times 2minthen the supernatant fluid was taken for use

223 Chip Processing Chip was put into the container withhydrochloric acid (100mmolL) oscillated for 4sim5min and

then washed with HPLC water put the chip into the biopro-cessor start with each hole plus 200120583L combined cushioningliquid and carry out oscillation for 5min at room tempera-ture discard liquid and then repeat the procedure then con-tinue with each hole plus 100 120583L serum sample at room tem-perature and carry out oscillation for 1 h Discard the liquidand wash the hole with 200120583L combining buffer 2 times(5min each time) and then with HPLC water Dismantle thechip perform natural drying and add 05 120583L SPA at eachpoint twice Read themeter reading chip withWCX2 proteinmicroarray after drying

224 Data Collection The same parameters were used forall the samples data were collected using Ciphergen Pro-teinChip 32 analysis software Set inspection sample laserintensity at 195 detection sensitivity at 8 collected dataquality load ratios (119872119885) in the range 10 times 103sim20times 103 andcollected position 20sim80 with each point collected 20 timesthe total points were 140 times Before each collection all-in-one protein microarray calibration instrument was usedto reduce the error to less than 01 Using the qualitycontrol protein array to do the repetitive detection the peakand strength variation coefficient (SVC) are controlled below005 and 15 respectively

225 Observing Indexes In the fingerprint qualitative loadratios (119872119885) between 11100 + HDa and 11900 + HDaexhibit cluster sample of fingerprint which is LGT existingperformance Observe LGT proteome abundance values andthe LGT dynamically in trauma group and control group

23 Statistical Analysis All measurement data was repre-sented withmean plusmn standard deviation (119909plusmnSD) andmultiplecomparisons of the average number of diversity samples wereexamined through SNK-q test the count data was examinedthrough 1205942 test SPSS110 was used for statistical analysis119875 lt 005 was considered statistically significant

3 Results

31 LGT Proteome Expression in TBI Patients The expressionof LGT proteome was119872119885 between 11100 + HDa and 11900+ HDa showing a group of three peaks or three peaks abovecluster of fingerprint The comparison between the upstreamand downstream peak cluster indicated the 119872119885 differenceof above 1000 quality units eye-catching background andoutstanding fingerprint characteristics Take the peak morethan 5 for LGT proteome positive diagnosis standard [8ndash10] In this study TBI patients showed significant LGT pro-teome expression patterns (Figure 2) while the control groupdid not present LGT protein expression spectrum (Figure 1)

32 The Comparison of Early Results after Admission amongthe GCS 6ndash8 Severe TBI Group GCS 3ndash5 Severe TBI Groupand the Control Group (Table 1) On the first day within 24hours of admission we collected the first serum specimento detect the differences of TBI patientsrsquo LGT proteomeabundance values The result shows that the LGT proteomeabundance of the GCS 6ndash8 severe TBI group and GCS 3ndash5

BioMed Research International 3

Table 1 The LGT proteome abundance comparison in the GCS 6ndash8 severe TBI group GCS 3ndash5 severe TBI group and normal controls

Group Cases (119899) Positive proportion Positive rate () LGT proteome abundance ()GCS 3ndash5 severe TBI group 44 3744 8409998771 1737 plusmn 687998771

GCS 6ndash8 severe TBI group 52 2152 3846 839 plusmn 538

Normal controls 35 035 0 179 plusmn 081Significant difference to normal controls (119875 lt 001) 998771Significant difference to severe brain injury group (119875 lt 001)

100

75

50

25

0

5000 10000 15000

Control 1

116597 + H116597 + H

Figure 1 LGT proteome does not present in the protein fingerprintspectra of normal controls

TBI1

0

25

50

75

100

5000 10000 15000

116254 + H

Figure 2 LGT proteome presents in protein fingerprint spectra ofTBI group

severe TBI group was obviously higher than of the controlgroup the LGT proteome abundance of the GCS 3ndash5 severeTBI groupwas significantly higher than of theGCS6ndash8 severeTBI group (Table 1)

33 LGT Proteome Results in Different Types of Cranio-cerebral Trauma According to the different types of TBIall the cases were assigned into A brain stem injury anddiffuse axonal injury B contusion and laceration of brainand (or) subduralintracerebral hematoma and C epiduralhematoma andor skull fractures analyzing three groups ofLGT proteome abundance values Results showed that theLGT proteome abundance values of groups A and B weresignificantly higher than of group C and the differencewas statistically significant (119875 lt 001) but there were nosignificant differences between groupsA andB (Table 2)

34 LGT Proteome Dynamic Change (Comparison with DeathGroup and Live Group) LGT proteome expression wasdetected in two groups of patients in the whole proceduredeath group was higher than the survival group in the courseof different stages then the abundance values of two groupsshowed a gradual decline rather than a progressive mannerBesides a moderate elevation was shown in both groups onthe seventh day During the early phase (days 1 and 4) LGTproteome abundance value of death group was significantly

Table 2 The analysis of the LGT proteome abundance in differentTBI groups

Group Cases (119899) LGT proteomeabundance

A brain stem injury and diffuseaxonal injury 35 1568 plusmn 1073998771

B contusion and laceration ofbrain and (or)subduralintracerebralhematoma

47 1416 plusmn 989998771

C epidural hematoma and (or)skull fracture 14 597 plusmn 513998771Compared with groupC (119875 lt 001)No significant difference with groupB (119875 gt 005)

higher than of the survival group (119875 lt 005) With theprogression of the course (days 7 and 14) there was nosignificant difference between the two groups (119875 gt 005)(Table 3)

4 Discussion

Traumatic brain injury (TBI) is the leading cause of death anddisability among young adults [2] Severe TBI is associatedwith a 30ndash70 mortality rate [14] The low sensitivity andspecificity of current indexes used to assess the injury degreeandprognosis (GCS score pupil reflection) and imaging (CT)contribute to the high morbidity and mortality of TBI [2 1516] Finding specific biomarker of proteome will provide anewmethod to explore the nosogenesis and prognosis of TBI[7] Proteome is the biomarker of TBI [17 18] The researchon the relationship between the dynamic change of serumand the injury degreeprognosis of TBI was common it oftencontained neuron-specific enolase (NSE) [2 15 19ndash22] S100B(S-100120573) [2 15 22ndash25] BB isozyme of creatine kinase (CK-BB) [2 15 26] glial fibrillary acidic protein (GFAP) [2 1522 27] myelin basic protein (MBP) [2 15] nerve growthfactor (NGF) [20] doublecortin (DCX) [20] high-mobilitygroup box 1 (HMGB1) [28] and so forth After severe TBIauthors of several studies have studied biomarkers in theacute phase biomarkers often contained S100B [14 29 30]ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) [31 32]glial fibrillary acidic protein (GFAP) [32] Interleukin-8 (IL-8) [33] Interleukin-10 (IL-10) [34] tumor necrosis factor 120572(TNF-120572) [33] cell-free DNA (cf-DNA) [35] and so forthHowever the correlation between structural damage andthese biomarkers has not been elucidated it was difficult tofind a serum biomarker with high specificity to assess theinjury degree and prognosis of severe TBI


Table 3 The Comparison of LGT proteome abundance in different pathogenetic stage

Prognosis Day 1 Day 4 Day 7 Day 14

Death group CasesLGT abundance

292609 plusmn 1092

231513 plusmn 579

161701 plusmn 1394

91192 plusmn 499

Survival group CasesLGT abundance

67898 plusmn 673

67607 plusmn 489

671053 plusmn 1368

67562 plusmn 707

119875 value lt001 lt005 gt005 gt005

Recently Pei et al [8ndash10] found that the serum con-centration of LGT proteome was related to the mortality oftumor patients through 146 cases the mortality was 100in the group which has persistent positive LGT proteomeconcentration and 0 in the group which was negativeIf LGT proteome is continuously negative it means thatthe disease lies stable On the contrary if LGT proteomeis continuously positive it implies disease progression andindicates poor prognosis Taken as a diagnosis platform topredict tumor progressing and by conducting correspondingtreatment it helps to extend the survival period of patientsso LGT proteome was a promising alert proteome whichcould forecast the prognosis of tumor patients NegativeLGT proteome fingerprint in the serum of patients withmalignant tumors could be viewed as the stationary phasewhile the positive LGT proteome could be viewed as theexacerbation stageWhen LGT proteome turns fromnegativeto positive it could be viewed as a sign of progression whiletransition from positive to negative indicates remission TheLGT proteome closely correlated with the severity of diseasecondition and prognosis in patients with critical illness [36]The LGT proteome presents in the serum of TBI patientsand it is a proteome that appeared under pathologic state[37] We presume that LGT proteome may also exist in theserum of severe TBI patients and could be prewarning for theaggravation and the mortality of severe TBI

Identification and purification of candidate biomarkers isa critical step in the biomarker development process sinceit provides insight into the disease biology and facilitates thedevelopment of analyte-specific assays [38] Technical prob-lems include selecting the best proteomics method for serumbiomarker discovery [5] Potential proteomic biomarkers canbe notably determined through SELDI-TOF-MS [39 40]SELDI-TOF-MS is a widely used technique for diagnosticbiomarker discovery in plasma serum and tissue and hasthe ability to carry out screening and identification Duringthe last years this technique has allowed high throughputand simplicity of experimental procedures to become animportant research tool for biomarker discovery [12 13] Thecharacteristic of LGT proteome is the appearance of a groupof three peaks or three peaks above cluster of fingerprint at119872119885 between 11100 + HDa and 11900 + HDa One of thespectrum abundance values was ≧20 but all abundancevalues were ≧5 compared with the upstream and down-stream peak cluster the 119872119885 values were more than 1000quality units with striking background as well as outstandingfingerprint characteristics So take the peakmore than 5 forLGT proteome positive diagnosis standard [8ndash10]

The experiment proceeded through SELDI-TOF-MStechnology the LGT proteome was detected in the proteomefingerprint with 100 relative abundance A fingerprintmarker just like a cluster appeared between 11100Da and11900Da119872119885 the upstream cluster peak got more than 1000mass units (119872119885) than downstream cluster peak it showedthe same result as the finding of Pei [8ndash10] so it proved thatthe LGT proteome could be detected in the serum of TBIpatients The LGT proteome was not detected in the serumof the controls The abundance of LGT proteome of GCS6ndash8 severe TBI group and GCS 3ndash5 severe TBI group wassignificantly higher than of the controls (119875 lt 001) Theabundance and positive rate of LGT proteome of GCS 3ndash5severe TBI group were significantly higher than of GCS 6ndash8 severe TBI group (119875 lt 001) Decreased serum LGT levelis associated with GCS scores The results showed that theLGTproteome could be detected in serumofTBI patients theabundance of LGT proteome is closely associated with patho-genesis of TBI patients TBI often involves a combination ofmechanical trauma and local hypoxemia on which serumbiomarker concentrations may provide critical therapeuticdata to evaluate the brain injury Serum biomarker after TBIcould be used as a source for the identification of the braininjury the extent of the injury and the time of its occurrenceand even for identifying its most likely outcome [41]

Our present study divided the TBI patients into threegroupsA brain stem injury and diffuse axonal injuryB con-tusion and laceration of brain and (or) subduralintracerebralhematoma andC epidural hematoma and (or) skull fractureThe result showed that the LGT proteome abundance ingroups A and B was significantly higher than in group C(119875 lt 001) but it did not reveal a significant differencebetween group A and group B The result showed that theLGT proteome had close relationship with the severity ofcraniocerebral trauma Neuronal and glial cell loss resultsfrom necrotic and apoptotic cell death during the primaryand secondary injury and may lead to an increase in variousneuron- and glia-specific proteins in serum [42] Elevatedserum levels of neuron- or glia-specific proteins indicateincreased permeability of the blood-brain barrier (BBB inaddition to neuronal and glial cell damage or loss) [33] SevereTBI not only causes substantial direct tissue damage butalso instantaneously breaks down existing biological barriersgenerating massive pathological responses (eg metabolicchanges) to toxic molecules and cellular debris Metabolicchanges include hypoxia altered cell metabolism (eg glu-cose utilization) disrupted energy levels and utilization(leading to ionic imbalance excitotoxicity etc) enhanced


inflammatory activity and increased systemic hormonal sec-retion [43 44] To our best knowledge the various formsof severe TBI can share common pathological ldquocomponentsrdquoduring both the primary and the secondary injury processesThere was no significant difference in the degree of injurybetween group A and group B which was consistent withtheir clinical symptoms due to the insignificant difference ofthe degree of injury between severe contusion and lacerationof brain and brain stem injury or diffuse axonal injury

The experiment also analyzed the LGT proteome abun-dance during the dynamic change of severe TBI patientsTheLGT proteome abundance was higher in death group thanin survival group in the initial pathologic stage and thendecreased in the two groups as the course was prolongedAt the early pathologic stage (days 1 and 4) the LGT pro-teome abundance was significantly higher in death groupthan in survival group (119875 lt 005) there was no significantdifference between the two groups in days 7 and 14 (119875 gt005) So the LGT proteome abundance at the early patho-genetic stage could be used to predict the different prognosisof TBI patients the higher the LGT proteome abundancewas the worse the prognosis was The experiment alsoshowed that the LGT proteome abundance of death andsurvival groups increased in day 7 It happened during thesecondary craniocerebral injury period so the increase of theLGT proteome abundance may be caused by the secondarycraniocerebral injury There is a second wave of long-lastingpathological changes called the secondary injurymechanismThese pathologic features include metabolic changes neu-roinflammation vascular abnormalities and neuronal andglial cell death [45 46] The temporal aspects of injurylike the onset of the various secondary pathologic features(cyto- and vasogenic factors leading to edema vasospasmand altered rates of perfusion) are especially important indynamically changing diseases like severe TBI This studyillustrates how to monitor the temporal pattern of changes(eg ldquotime to peakrdquo) of serumLGT levels which can be usefulfor identifying injury severity and outcomeWe can state thatthe determination of LGT levels in serum could act as a bio-marker for the early prediction of mortality after severe TBI

5 Conclusion

This is the first study to investigate the biokinetics of LGTproteome a potential biomarker of severe TBI in serumof severe TBI patients The LGT proteome presents in theserum of severe TBI patients The abundance diversity ofLGTproteome is closely associatedwith pathologic conditionof TBI patients Serum LGT proteome may be used asa promising marker for evaluating severity of severe TBIFurther studies of LGT proteome are needed to validate thesefindings in a larger scale of samples and should be conductedto support these associations with outcomes

The Innovation

Serum LGT proteome was detected by surface enhancedlaser desorptionionisation time-of-flight mass spectrometer(SELDI-TOF-MS) The LGT proteome presents in the serum

of severe TBI patients The abundance diversity of LGT pro-teome is intimately associated with pathogenetic condition ofTBI patients Serum LGT proteome may be used as a pro-mising marker for evaluating severity of severe TBI

Conflict of Interests

The topics presented herein are not covered by any manufac-turer and the relevant employer or other economic organiza-tions or direct or indirect economic interests of sponsors

Authorsrsquo Contribution

Hongming Ji and Changchen Hu contributed equally to thiswork and should be considered co-first authors

Acknowledgments

This research was funded by Medical Science and Tech-nology Research projects of Health Department of HubeiProvince (JX4B42) and by the National Science Foundationof China (30901774) and National Science Foundation ofShanxi (2014011038-2)

References

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[2] G W Hergenroeder J B Redell A N Moore and P KDash ldquoBiomarkers in the clinical diagnosis andmanagement oftraumatic brain injuryrdquo Molecular Diagnosis and Therapy vol12 no 6 pp 345ndash358 2008

[3] S D Timmons ldquoAn update on traumatic brain injuriesrdquo Journalof Neurosurgical Sciences vol 56 no 3 pp 191ndash202 2012

[4] P M Kochanek H Bramlett W D Dietrich et al ldquoA novelmulticenter preclinical drug screening and biomarker consor-tium for experimental traumatic brain injury operation braintrauma therapyrdquo The Journal of TraumamdashInjury Infection ampCritical Care vol 71 no 1 supplement pp S15ndashS24 2011

[5] D V Agoston A Gyorgy O Eidelman and H B Pollard ldquoPro-teomic biomarkers for blast neurotrauma targeting cerebraledema inflammation and neuronal death cascadesrdquo Journal ofNeurotrauma vol 26 no 6 pp 901ndash911 2009

[6] D V Agoston and M Elsayed ldquoSerum-based protein biomark-ers in blast-induced traumatic brain injury spectrum disorderrdquoFrontiers in Neurology vol 3 article 107 Article ID Article 1072012

[7] D A Prieto X Ye and T D Veenstra ldquoProteomic analysisof traumatic brain injury the search for biomarkersrdquo ExpertReview of Proteomics vol 5 no 2 pp 283ndash291 2008

[8] Y Pei S Guo Q Wang et al ldquoThe first observation of theclinical significance of LGT proteome for cancer patientsrdquoCancer Research and Clinic vol 17 no 3 pp 156ndash158 2005

[9] P Yi S Guo Q Wang et al ldquoA newly discovered fingerprintof functional proteomerdquo Zhongliu Yanjiu Yu Lingchuang vol 17no 3 pp 153ndash155 2005

[10] P Yi ldquoDiscovery and concluding of characteristics of prewarn-ing proteome fingerprints which mark prognostic of malignant


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[11] J R Ren G L Zhang H M Ji et al ldquoThe clinical signifiancechange of serumLGT proteome in patients with traumatic braininjury a pilot studyrdquo Zhongguo Yaowu Yu Lingchuang vol 10no 10 pp 1095ndash1097 2010

[12] T Gemoll U J Roblick G Auer H Jornvall and J K Haber-mann ldquoSELDI-TOF serum proteomics and colorectal cancer acurrent overviewrdquo Archives of Physiology and Biochemistry vol116 no 4-5 pp 188ndash196 2010

[13] V Seibert M P A Ebert and T Buschmann ldquoAdvances inclinical cancer proteomics SELDI-ToF-mass spectrometry andbiomarker discoveryrdquo Briefings in Functional Genomics andProteomics vol 4 no 1 pp 16ndash26 2005

[14] A B da Rocha R F Schneider G R de Freitas et al ldquoRole ofserum S100B as a predictive marker of fatal outcome followingisolated severe head injury or multitrauma in malesrdquo ClinicalChemistry and Laboratory Medicine vol 44 no 10 pp 1234ndash1242 2006

[15] S J I Sandler A A Figaji and P D Adelson ldquoClinical appli-cations of biomarkers in pediatric traumatic brain injuryrdquoChildrsquos Nervous System vol 26 no 2 pp 205ndash213 2010

[16] J E Risdall and D K Menon ldquoTraumatic brain injuryrdquoPhilosophical Transactions of the Royal Society B BiologicalSciences vol 366 no 1562 pp 241ndash250 2011

[17] K KWWang A K Ottens M C Liu et al ldquoProteomic identi-fication of biomarkers of traumatic brain injuryrdquo Expert Reviewof Proteomics vol 2 no 4 pp 603ndash614 2005

[18] P M Kochanek R P Berger H Bayir A K Wagner L WJenkins andR S B Clark ldquoBiomarkers of primary and evolvingdamage in traumatic and ischemic brain injury diagnosisprognosis probingmechanisms and therapeutic decisionmak-ingrdquo Current Opinion in Critical Care vol 14 no 2 pp 135ndash1412008

[19] J Li H C Liu G L Zhang et al ldquoDetermination of serumneuron-specific enolase (NSE) in patients with severe traumaticbrain injuryrdquoActaMedicinae Universitatis Scientiae et Technolo-giae Huazhong vol 40 no 2 pp 225ndash228 2011

[20] A Chiaretti G Barone R Riccardi et al ldquoNGF DCX andNSE upregulation correlates with severity and outcome of headtrauma in childrenrdquoNeurology vol 72 no 7 pp 609ndash616 2009

[21] CWoertgen R D Rothoerl and A Brawanski ldquoNeuron-speci-fic enolase serum levels after controlled cortical impact injuryin the ratrdquo Journal of Neurotrauma vol 18 no 5 pp 569ndash5732001

[22] A E Bohmer J P Oses A P Schmidt et al ldquoNeuron-specificenolase S100B and glial fibrillary acidic protein levels as out-come predictors in patients with severe traumatic brain injuryrdquoNeurosurgery vol 68 no 6 pp 1624ndash1630 2011

[23] S M Bloomfield J McKinney L Smith and J BrismanldquoReliability of S100B in predicting severity of central nervoussystem injuryrdquoNeurocritical Care vol 6 no 2 pp 121ndash138 2007

[24] E Kovesdi J Luckl P Bukovics et al ldquoUpdate on proteinbiomarkers in traumatic brain injury with emphasis on clinicaluse in adults and pediatricsrdquo Acta Neurochirurgica vol 152 no1 pp 1ndash17 2010

[25] A Sofijanova K Piperkova and D Al Khalili ldquoPredicting out-come after severe brain injury in risk neonates using the serumS100B biomarker results using single (24h) time-pointrdquo Prilozivol 33 no 1 pp 147ndash156 2012

[26] M E Carr Jr L N Masullo J K Brown and P C LewisldquoCreatine kinase BB isoenzyme blood levels in trauma patientswith suspected mild traumatic brain injuryrdquoMilitary Medicinevol 174 no 6 pp 622ndash625 2009

[27] L Schiff N Hadker S Weiser and C Rausch ldquoA literaturereview of the feasibility of glial fibrillary acidic protein as abiomarker for stroke and traumatic brain injuryrdquo MolecularDiagnosis and Therapy vol 16 no 2 pp 79ndash92 2012

[28] K-Y Wang G-F Yu Z-Y Zhang Q Huang and X-Q DongldquoPlasma high-mobility group box 1 levels and prediction ofoutcome in patients with traumatic brain injuryrdquo Clinica Chi-mica Acta vol 413 no 21ndash23 pp 1737ndash1741 2012

[29] A Rodrıguez-Rodrıguez J J Egea-Guerrero A Leon-Justelet al ldquoRole of S100B protein in urine and serum as an earlypredictor of mortality after severe traumatic brain injury inadultsrdquo Clinica Chimica Acta vol 414 pp 228ndash233 2012

[30] T Rainey M Lesko R Sacho F Lecky and C Childs ldquoPredict-ing outcome after severe traumatic brain injury using the serumS100B biomarker results using a single (24h) time-pointrdquoResuscitation vol 80 no 3 pp 341ndash345 2009

[31] S Mondello A Linnet A Buki et al ldquoClinical utility of serumlevels of ubiquitin C-terminal hydrolase as a biomarker forsevere traumatic brain injuryrdquo Neurosurgery vol 70 no 3 pp666ndash675 2012

[32] S Mondello L Papa A Buki et al ldquoNeuronal and glial markersare differently associated with computed tomography findingsand outcome in patients with severe traumatic brain injury acase control studyrdquoCritical Care vol 15 no 3 article R156 2011

[33] D M Stein A L Lindel K R Murdock J A Kufera JMenaker and TM Scalea ldquoUse of serum biomarkers to predictsecondary insults following severe traumatic brain injuryrdquoShock vol 37 no 6 pp 563ndash568 2012

[34] F M S Soares N M de Souza M L Schwarzbold et alldquoInterleukin-10 is an independent biomarker of severe trau-matic brain injury prognosisrdquo NeuroImmunoModulation vol19 no 6 pp 377ndash385 2012

[35] H Macher J J Egea-Guerrero J Revuelto-Rey et al ldquoRoleof early cell-free DNA levels decrease as a predictive markerof fatal outcome after severe traumatic brain injuryrdquo ClinicaChimica Acta vol 414 pp 12ndash17 2012

[36] J-R Ren H-M Ji J-M Ji J-H Feng G-L Zhang and H-W Zhang ldquoThe clinical significance of the relationship betweenserum lost goodwill target proteome and the acute physiologyand chronic health evaluation II score of patient with criticalillnessrdquo Zhongguo Wei Zhong Bing Ji Jiu Yi Xue vol 23 no 3pp 134ndash137 2011

[37] J R Ren G L Zhang H M Ji et al ldquoThe clinical significanceof the serum LGT proteome for patients with traumatic braininjuryrdquoZhonghua ShiyuanWaike Zhazhi vol 28 no 6 pp 1001ndash1002 2011

[38] A L Bulman and E A Dalmasso ldquoPurification and identifica-tion of candidate biomarkers discovered using SELDI-TOFMSrdquoMethods in Molecular Biology vol 818 pp 49ndash66 2012

[39] S C Karatayli S G Alagoz D Mizrak et al ldquoPotential pro-teomic biomarkers in assessing liver fibrosis using SELDI-TOFMSrdquo Turkish Journal of Gastroenterology vol 23 no 1 pp 46ndash53 2012

[40] P Li D Zhang and C Guo ldquoSerum biomarker screening forthe diagnosis of early gastric cancer using SELDI-TOF-MSrdquoMolecular Medicine Reports vol 5 no 6 pp 1531ndash1535 2012

[41] S Menascu A Brezner S M Tshechmer and P G RumenyldquoSerum biochemical markers for brain damage in children with


emphasis on mild head injuryrdquo Pediatric Neurosurgery vol 46no 2 pp 82ndash88 2010

[42] B A Stoica andA I Faden ldquoCell deathmechanisms andmodu-lation in traumatic brain injuryrdquoNeurotherapeutics vol 7 no 1pp 3ndash12 2010

[43] A M Cook A Peppard and B Magnuson ldquoNutrition consid-erations in traumatic brain injuryrdquoNutrition in Clinical Practicevol 23 no 6 pp 608ndash620 2008

[44] J-F Feng X Zhao G G Gurkoff K C Van K Shahlaie andB G Lyeth ldquoPost-traumatic hypoxia exacerbates neuronal celldeath in the hippocampusrdquo Journal of Neurotrauma vol 29 no6 pp 1167ndash1179 2012

[45] I Cernak and L J Noble-Haeusslein ldquoTraumatic brain injuryan overview of pathobiology with emphasis on military popu-lationsrdquo Journal of Cerebral Blood Flow andMetabolism vol 30no 2 pp 255ndash266 2010

[46] J J Donkin and R Vink ldquoMechanisms of cerebral edemain traumatic brain injury therapeutic developmentsrdquo CurrentOpinion in Neurology vol 23 no 3 pp 293ndash299 2010

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Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Computational and Mathematical Methods in Medicine

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


time-of-flight mass spectrometry (SELDI-TOF-MS) [12 13]technology which can detect the exact LGT concentration inthe serum of TBI severe patients

2 Materials and Methods

21 General Data 96 TBI cases treated in our departmentbetween March 2006 and July 2009 were analyzed retrospec-tively There were 77 males and 19 females with age range of16ndash83 years (mean age 317plusmn263) interval time from injuryto clinical (h) 05sim115 h average 71 plusmn 22 Glasgow ComaScale (GCS) score 44 cases with 3sim5 points 52 cases with 6sim8points and 32 cases receiving emergency operation traumacause 46 cases by traffic accident 22 cases by injury due tofall or mine disaster 19 cases by strike injury and 6 caseswith other causes major trauma type 47 cases with contu-sion and laceration of brain and (or) subduralintracerebralhematoma 14 cases with epidural hematoma and 35 caseswith diffuse axonal injury and brain stem injury All caseswere diagnosed as craniocerebral trauma by cranial CTscanning after the admission 69 survival cases carried onprognostic evaluation with Glasgow outcome score (GOS)6 months later The exclusion criteria were as follows beingunder 16 years old death in 24 hours from admission severeinfection or combined injury malignant tumor or autoim-mune disease with damage of important organs especiallyliver and kidney (multiple organ failure) and transfusionof blood in operation Meanwhile 35 healthy cases werechosen as control group This project has been reviewed bythe Medical Ethics Committee of Shanxi Provincial PeoplersquosHospital At the same time signed informed consent ofresearch projects of every patient with patients themselvesor their guardians was obtained The factors such as generalcondition age and sex were comparable (in research groupand the normal controls) between the two groups (119875 gt 005)

22 SELDI-TOF-MS Detection

221 Main Instruments and Reagents Themain instrumentsand reagents included protein fingerprint meter WCX2chips PBS-IIc chip reading machine and Ciphergen Pro-teinChip 32 analysis software urea sodium acetate [NaAc(100mmolL NaAc pH 40)] HPLC water and SPA

222 Serum Sample Processing 5mL sera were taken inthe morning when patients were hollow and were putin VACVTTE vacuum quantitative mining container andplaced at room temperature for 60min (including the timewhen samples were out of the body) and then the serumwas separated from blood at 3000 g times 10min and preservedat minus80∘C serum samples were thawed and put in ice bath(10000 g times 2min) take 5 120583L serum sample and add 10 120583L urea(9mmolL) The sample underwent oscillation for 30minfollowed by the addition of 180 120583L buffer (100mmolL NaAcpH = 40) at 4∘C and was centrifuged at 10000 g times 2minthen the supernatant fluid was taken for use

223 Chip Processing Chip was put into the container withhydrochloric acid (100mmolL) oscillated for 4sim5min and

then washed with HPLC water put the chip into the biopro-cessor start with each hole plus 200120583L combined cushioningliquid and carry out oscillation for 5min at room tempera-ture discard liquid and then repeat the procedure then con-tinue with each hole plus 100 120583L serum sample at room tem-perature and carry out oscillation for 1 h Discard the liquidand wash the hole with 200120583L combining buffer 2 times(5min each time) and then with HPLC water Dismantle thechip perform natural drying and add 05 120583L SPA at eachpoint twice Read themeter reading chip withWCX2 proteinmicroarray after drying

224 Data Collection The same parameters were used forall the samples data were collected using Ciphergen Pro-teinChip 32 analysis software Set inspection sample laserintensity at 195 detection sensitivity at 8 collected dataquality load ratios (119872119885) in the range 10 times 103sim20times 103 andcollected position 20sim80 with each point collected 20 timesthe total points were 140 times Before each collection all-in-one protein microarray calibration instrument was usedto reduce the error to less than 01 Using the qualitycontrol protein array to do the repetitive detection the peakand strength variation coefficient (SVC) are controlled below005 and 15 respectively

225 Observing Indexes In the fingerprint qualitative loadratios (119872119885) between 11100 + HDa and 11900 + HDaexhibit cluster sample of fingerprint which is LGT existingperformance Observe LGT proteome abundance values andthe LGT dynamically in trauma group and control group

23 Statistical Analysis All measurement data was repre-sented withmean plusmn standard deviation (119909plusmnSD) andmultiplecomparisons of the average number of diversity samples wereexamined through SNK-q test the count data was examinedthrough 1205942 test SPSS110 was used for statistical analysis119875 lt 005 was considered statistically significant

3 Results

31 LGT Proteome Expression in TBI Patients The expressionof LGT proteome was119872119885 between 11100 + HDa and 11900+ HDa showing a group of three peaks or three peaks abovecluster of fingerprint The comparison between the upstreamand downstream peak cluster indicated the 119872119885 differenceof above 1000 quality units eye-catching background andoutstanding fingerprint characteristics Take the peak morethan 5 for LGT proteome positive diagnosis standard [8ndash10] In this study TBI patients showed significant LGT pro-teome expression patterns (Figure 2) while the control groupdid not present LGT protein expression spectrum (Figure 1)

32 The Comparison of Early Results after Admission amongthe GCS 6ndash8 Severe TBI Group GCS 3ndash5 Severe TBI Groupand the Control Group (Table 1) On the first day within 24hours of admission we collected the first serum specimento detect the differences of TBI patientsrsquo LGT proteomeabundance values The result shows that the LGT proteomeabundance of the GCS 6ndash8 severe TBI group and GCS 3ndash5






100

75

50

25

0

5000 10000 15000

Control 1

116597 + H116597 + H


TBI1

0

25

50

75

100

5000 10000 15000

116254 + H









47 1416 plusmn 989998771



4 Discussion






292609 plusmn 1092

231513 plusmn 579

161701 plusmn 1394

91192 plusmn 499


67898 plusmn 673

67607 plusmn 489

671053 plusmn 1368

67562 plusmn 707

119875 value lt001 lt005 gt005 gt005








5 Conclusion


The Innovation







Acknowledgments


References
























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















100

75

50

25

0

5000 10000 15000

Control 1

116597 + H116597 + H


TBI1

0

25

50

75

100

5000 10000 15000

116254 + H









47 1416 plusmn 989998771



4 Discussion






292609 plusmn 1092

231513 plusmn 579

161701 plusmn 1394

91192 plusmn 499


67898 plusmn 673

67607 plusmn 489

671053 plusmn 1368

67562 plusmn 707

119875 value lt001 lt005 gt005 gt005








5 Conclusion


The Innovation







Acknowledgments


References
























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




















292609 plusmn 1092

231513 plusmn 579

161701 plusmn 1394

91192 plusmn 499


67898 plusmn 673

67607 plusmn 489

671053 plusmn 1368

67562 plusmn 707

119875 value lt001 lt005 gt005 gt005








5 Conclusion


The Innovation







Acknowledgments


References
























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















5 Conclusion


The Innovation







Acknowledgments


References
























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















research article determination of serum lost goodwill...

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