MINISTRY OF EDUCATION AND SCIENCE OF THE RUSSIAN

FEDERATION

Ulyanovsk State University

Institute for Medicine, Ecology and Physical Education

T`Biktimirov`sMedicineFaculty

T.P.Gening, T.V.Abakumova, S.O.Gening

WORKSHOP ON PHYSIOLOGY OF VISCERAL SYSTEM

Methodological guidancefor students of medical faculty

Ulyanovsk, 2019

2

УДК 612.8

ББК 28.903я73

G 36

Printed by the decision of the Academic Council

of Institutefor Medicine, Ecology and Physical EducationUlSU

(document№ 2/212, 16.10.2019)

Rewievers:

Associate Professor of the Department of

MorphologyIME&PhEUlSUPh.DT.I.Kuznetsova

Head of gynecology department, MD, doctor of the highest qualification category

Ulyanovsk Regional Clinical Oncology CenterI.I.Antoneeva

Gening T.P.

G36 Workshop on physiology of visceral system:methodological guidancefor

students of medical faculty. / T.P.Gening, T.V.Abakumova, S.O.Gening. – Ulyanovsk: UlSU,

2019. – 25 p.

The methodical manual presents the course of work in the classroom classes,

questions for colloquiums in physiology of visceral system for foreign medical

students as materials for preparing for practical classes and independent work. The

manual provides the formation of the necessary competencies provided for by the

Federal State Educational Standard.

© Gening T.P., Abakumova T.V.,

Gening S.O. 2019

©Ulyanovsk State University, 2019

3

CONTENTS

1Laboratory research Heart physiology. Properties of a cardiac muscle . . . . . . . . . . . . . . . . . . . 4

2 Laboratory research Heart physiology.The cardiac cycle.

Regulation of heart work. Methods of heart work research. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

3 Laboratory research Overview of the circulation.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7

4 Laboratory research Questionsto the Colloquium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..9

5 Laboratory research Breath physiology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

6 Laboratory research Questionsto the Colloquium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

7 Laboratory research Digestion physiology. Digestion in a mouth and in a stomach . . . . . . . . 11

8 Laboratory research Digestion physiology.Digestion in intestines.

A role of a pancreas and liver in digestion.Motility of gastrointestinal tract (GI).

Absorption in various departments of a GI. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

9 Laboratory research Questionsto the Colloquium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14

10 Laboratory research Allocation physiology. Regulation of kidneyfunctions. . . . . . . . . . . . . 15

11 Laboratory research Physiology of endocrine glands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

12 Laboratory research Metabolism. Thermal control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16

13 Laboratory research Questionsto the Colloquium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

14 Laboratory research Blood physiology. Physiology of erythrocytes . . . . . . . . . . . . . . . . . . . .20

15 Laboratory research Physiology of leukocytes and thrombocytes. Hemolysis. . . . . . . . . . . . 22

16 Laboratory research Blood fibrillation. The doctrine about blood groups. . . . . . . . . . . . . . . .23

17 Laboratory research Questionsto the Colloquium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25

18 Laboratory research Practical Skills . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

4

Laboratory research number 1

Heart physiology. Properties of a cardiac muscle

Work 1. Monitoring of a cardiac cycle of the frog and graphic registration of the heart

beat. The effect of electrical stimuli application on the cardiac activity Objective: demonstration of the phases of the cardiac cycle of the frog heart and the evolution of

its excitability by the using the graphical method.

The object of study: the frog. Equipment: universal tripod, serphin, dissecting kit, Ringer solution, amplifier, traductor

Stages: 1. The graphical recording of the normal mechanical activity of the heart

2. Thegraphical recording of the effect of electrical stimuli on the mechanical activity of the

heart, first in systole and then in diastole

Results: the normal cardiogram is sinusoid and we can distinguish the two phases of the cardiac

revolution: systole, diastole. By applying the experimental electrical stimuli we obtain different

responses according the phases of the cardiac cycle in which the frog`s heart is found:

extrasystole which is followed inevitably by a prolonged resting period.

Conclusion: in each systole the myocardium is unexcitable. In diastole the myocardium

becomes excitable and if there is an artificial stimuli the response will be an extra-systole. Any

extrasystole is constantly followed by a prolonged resting period

Work 2. The Stannius ligatures Objective: demonstration of the mechanism of the conduction system in the heart The object of study: heart of frog.

Equipment: universal tripod, serphin, ligatures, Ringer solution.

Stages: The practical session consist in “in situ” applying of some ligatures in different areas of the heart

to demonstrate their effect on the cardiac activity.

I. Apply a thread around the S-A node to separate the Remak ganglion from the rest of

the heart

II. Apply a thread around the S-A node and another one around the A-V node III. Remove Ist and IInd ligatures and apply IIIrd ligature to the apex of the heart.

Results: The parts of the heart The number of beats for 10 s

Source value Afterapplying 1 ligature 2 ligatures

Venoussinus Atrium Ventricle

Conclusion:determined and compared the levels of automation of various parts of the

conduction system of the heart.

Laboratory research 2

Heart physiology.The cardiac cycle.Regulation of heart work. Methods of heart work

research

Work 1. Recording and analysis of electrocardiogram

Objective: to familiarize with the recording technique and the basics of analysis.

The object of the study: human

Equipmentelectrocardiograph, a 10% solution of sodium chloride.

Stages:

5

1. Registration of ECG:

A) ground the electrocardiograph, include it in the network 220V;

B) the test subject is laid on the couch, applyelectrodes in accordance with the adopted scheme

for recording ECG from limb leads. When applying electrodes, use wipes moistened with saline

solution;

C) record the calibration signal (1 mV = 10 mm);

D) record the ECG at 1, 2, 3 limb leads to a visual acquaintance, at minimum speed;

E) at the speed of 50 mm/s record the ECG in the 2nd lead for analysis.

2. The ECG analysis:

A) pay attention to the presence of waves, their severity (measure amplitude of P, R, T),

orientation, shape, position;

B) measure the duration of intervals PQ, QRST, QRS, RR;

C) calculate the heart rate;

D) evaluate the systolic indicator.

Results: (electrocardiogram and table with data analysis)

Parameter Duration, sec Amplitude, mV

norm experience norm experience

P wave 0,06-0,11 <0,25

PR interval

0,12-0,20

PQsegment

0,08

QRS complex

<0,12 0,8-1,2

QTinterval

0,36-0,44

6

STsegment

0,12

Twave 0,16 <0,5

Conclusion:Registered and decoded ECG

Work 2.Listening to heart tones.

Objective: to determine heart sounds, heart borders, apical impulse.

Equipment:phonendoscope

Stages:

The external manifestations of human heart activity include: the apical impulse, heart tones and

electrical phenomena that occur in the working heart. The apical impulse is the rhythmic protrusion of the

anterior wall of the chest that occurs during contraction of the heart. In an adult, it is localized to the left

in the 5th intercostal space, 1.0-1.5 cm medially from the mid-clavicular line.

The apical impulse is determined by visual inspection, palpation and the method of graphic recording.

External examination establish the place of rhythmic protrusion of the anterior chest wall and the size of

this protrusion.Putting the palm of the right hand to the chest in the area of the apical impulse, determine

it (position, strength and area of impact).

Heart tones are determined by auscultation using a stethoscope (or phonendoscope) in the best listening

places: I tone is heard better in the area of the apical impulse (the place of the best audible work of the

mitral valve) and the xiphoid process of the sternum (the place of the best audibility of the tricuspid

valve); II tone is best heard in the 2nd intercostal space to the left of the sternum (the place of the best

audibility of the pulmonary valves) and the rim of it (the place of the best audibility of the aortic valves).

Determining the boundaries of the

heart is produced by percussion, most often through the impact of an

impression with a bent middle finger

of the right hand on the middle finger of the left hand located on the body

part to be percussed. During

percussion, there is either a dull sound, if the area to be punctured has a dense

consistency (for example, heart, liver),

or a clear sound, if there is air in the

area to be percussed (for example, lungs). The difference of these sounds

set the boundaries of the body.

Determining the boundaries of the heart begins on the right side. First

percussion from above on the right

mid-clavicular line to identify a dull

sound (liver). The finger is then carried 2-3 cm above this boundary

and percussed, moving the finger

towards the sternum until a dull sound appears. The right border of the heart

extends 1 cm to the right of the right

edge of the sternum at the level of the 4th intercostal space.

The upper boundary is determined by percuting along the left mid-subclavial line, dropping from the first

intercostal space. Normally, it is located in the 3rd intercostal space.

7

The left border is determined by percuting along the 4-5th intercostal space, depending on the location of

the apical impulse, from the middle axillary line towards the sternum. The left border coincides with the

region of the apical impulse, i.e. it is located 1-1.5 cm inside from the mid-clavicular line.

These boundaries characterize relative cardiac dullness and give an idea of the true dimensions of the

heart.

Conclusion:Assess the nature of the apical impulse, heart tones and explain their nature.

Work 3.Determination of the duration of the cardiac cycle by pulse.

Objective: to learn how to determine the duration of the cardiac cycle by pulse.

Equipment: stopwatch.

Stages: Grop for a pulse on the radial artery at home or a colleague. Count the number of pulse beats for

5 s several times within 3 minutes. 5 divided by each found number, calculate the average duration of the

cardiac cycle in every 5 seconds.

Then determine the number of pulse beats for 1 min. 60 divided by the number found and find the

average duration of the cardiac cycle.

Conclusion: Determine whether the method for determining the duration of the cardiac cycle by

fractional pulse counting (every 5 s) has an advantage over the counting technique per 1 minute.

Laboratory research 3

Overview of the circulation

Work 1.Blood pressure measurement in humans by the method of Riva-Rocci

Objective: to learn the technique of measuring blood pressure by palpation method of Riva-

Rocci, to correlate the obtained results with the norm.

The object of the study: human.

Equipment: sphygmomanometer.

Stages:

1. The test subject sits down. Put at his naked shoulder the cuff so that it will fit tightly over the

shoulder, but not tight tissues. The tube of cuff connect with the sphygmomanometer.

2. Palpate the pulse at the radial artery, pump up the air in the cuff up to its disappearance.

3. Slowly release air from the cuff and note the pressure when pulse appears.

Results: (the values of systolic pressure)

Conclusion: We learned how to measure blood pressure by the method of Riva-Rocci

8

Work 2.Blood pressure measurement in humans by the method of Korotkov

Objective: to master the auscultatory method of blood pressure measurement by Korotkov’s

method, to correlate the obtained results with the norm.

The object of the study: human.

Equipment: sphygmomanometer, phonendoscope.

Stages:

1. The test subject sits down. Put at his naked shoulder the cuff (1) and connect it with the

sphygmomanometer (2,3).

2. Set the phonendoscope in the cubital fossa over the brachial artery (4).

3. Create the pressure in the cuff above the estimated systolic pressure.

4. Slowly release air from the cuff, and listen to the Korotkov’s tones. Note pressure, when tones

appear and disappear.

Results: (the values of systolic and diastolic pressure)

Conclusion: We learned how to measure blood pressure by the method of Korotkov

Work 3.The study of the pulse

Objective: to master the technique of palpation of the pulse and to know its main characteristics.

The object of the study: human

Stages:

Four fingers of the right hand place on the wrist on the projection of the radial artery and assess:

A) pulse rate by counting the number of oscillations for 15 s;

B) rhythm by triple counting the number of oscillations for 15 seconds;

B) degree of filling by the amplitude of the vibration;

G) the degree of stress by the force with which you need compress the artery to stop the

oscillations;

D) speed by rise and fall times of the pulse wave.

Results:

Thepulsecharacteristics The evaluation of the pulse Frequency (beats/min) Rhythm (beats/15 s)

Filling Voltage

Thespeed

9

Laboratory research 4

Questionsto the Colloquium

1. Excitability of a cardiac muscle

2. Contractility of a cardiac muscle

3. Conductivity of a cardiac muscle

4. Automaticity

5. Self-regulation of heart work

6. Nervous regulation of heart work

7. Heart reflexes.

8. Humoral influences on heart work

9. The cardiac cycle

10. Tones of heart. Phonocardiography.

11. Electrocardiography.

12. Physical characteristics of the blood circulatory system:

1) Functional parts of the circulation.

2) The laws of hemodynamics.

13. Blood pressure.

1) Pressure in the various portions of the circulation.

2) Clinical methods for measuring systolic and diastolic pressure.

3) Regulation of arterial pressure.

14. Arterial pressure pulsation.

15. Venous pressure pulsation.

16. The microcirculation.

1) Structure of the microcirculation and capillary system.

2) Average function of the capillary system.

3) Fluid filtration across capillaries.

17. Control of blood flow by the tissues.

Laboratory research 5

Breath physiology

Work 1.Spirometry

Objective: to investigate the characteristics of external respiration, to give an opinion on the

conformity of the received data to the existing standards.

The object of the study: human Equipment: a water spirometer or volumespirometer, dry spirometer, alcohol, cotton wool.

Stages: 1. Evaluation of vital capacity of the lungs:

A) set a spirometer so that the baseline corresponded to 0. When using dry spirometer, combine

0 on the scale to the arrow.

b) after taking a maximum deep breath from the surrounding air, make the maximum exhalation

into the spirometer;

B) note the level corresponding to the VC value. 2. Evaluation of inspiratory reserve volume:

A) fill the spirometer with air to the level of 4 L; B) after quiet inhalation from the surrounding air, make the maximum inhalation from the

spirometer. Calculate the inspiratory reserve volume as the difference of the first and second

readings of the spirometer.

10

Note: not evaluate IRV with volumespirometer.

3. Evaluation of expiratory reserve volume: After a quiet exhalation in the environment, make the maximum exhalation into the spirometer.

4. Evaluation of breathing capacity: Exercise 5 alternating inhalations from the surrounding air and exhalations into the spirometer.

Evaluate the mean value of breathing capacity (BC). 5. The calculation of the ratio of pulmonary ventilation:

RPV = (BC – harmful air space) / (residual air + ERV)

Results:

BC IRV ERV Theirsum VC

Conclusion:The spirometry method was tested

Work 2.Pulmonary function. Volumes and Capacities

Objective: to observe experimentally, record and calculate selected pulmonary volumes and

capacities.

The object of the study: human

Equipment: BIOPAC Student Lab, alcohol, cotton wool.

Stages: 1. Validate Calibration. Cycle the AFT6 syringe plunger in and out completely 5 times

(10strokes)

2. Click Record. Breathe normally forn5 breaths. Inhale as deeply as you can. Exhale as

deeply as you can. Breathe normally for 5 breaths.

Results: Review the measurements described in the introduction to identify the appropriate selected

area for each: total lung capacity, tidal volume, inspiratory reserve volume, expiratory

reserve volume, vital capacity, expiratory capacity, inspiratory capacity, functional residual

capacity, residual volume

Conclusion:In this laboratory work we used an airflow transducer and software was convert

airflow to volume, thus approximating the volume reading of a spirometer.

The resultant record of volume change called a spirogram.

Work 3.Calculation of dynamic respiratory parameters

Objective:Calculate dynamic respiratory rates

The object of the study: human

Equipment: water or dry spirometers, mouthpieces for personal use, cotton, alcohol.

Stages:

1. Forced vital capacity of the lungs (FVC) - the amount of air that can be exhaled with a

forced exhalation after maximum inhalation. After maximal slow inhalation from the

atmosphere, perhaps make a quick maximum exhalation into the spirometer. Compare your

actual VC to FVC.

Normally, the difference between VC and FVC is 100–300 ml. Increasing this difference to 1500

ml or more indicates resistance to air flow due to narrowing of the lumen of the small bronchi.

The duration of the fastest possible expiration ranges from 1.5 to 2.5 s.

2. Anatomical dead space (ADS) - the volume in which gas exchange does not occur - air

nasopharynx, trachea, large bronchi; no direct definition. Take ADS = 150 ml.

3. Respiratory rate (RR) - the number of respiratory cycles (inhale — exhale) in 1 min. Count the

number of breathing cycles per minute.

4. The minute respiratory volume (MRV) - the amount of air ventilated in the lungs for 1 minute.

MRV = RV x RR

5. Alveolar ventilation - the volume of exhaled air entering the alveoli.

AV = 66-80% of the MRV

6. Respiratory reserve - an indicator characterizing the possibility of increasing ventilation.

11

Respiratory reserve = MRV - MRV

Normally, this difference is 85% of MRV

Results:

Indicators Results Regulatory Values

FVC 3 000 – 7 000 l

ADS 150 ml

RR 16 – 18 inminute

MRV 8 – 12 l

Alveolar ventilation 0,8 l/min

Respiratory reserve 85 %

Conclusion: We learned to calculate dynamic respiratory rates.

Laboratory research 6

Questionsto the Colloquium

1. Breath definition. Breath stages.

2. Mechanism of breath and exhalation.

3. Pressure in a pleural cavity. Pheumothorax.

4. Pulmonary volumes. Spirometry, spirography, pneumotachography.

5. Composition of the inhaled, exhaled and alveolar air.

6. Gas exchange in lungs.

7. Blood transport of gases.

8. Gas exchange in tissue.

9. Respiratory center (Rhythm generator). Automaticity of the respiratory center.

10. Protective respiratory reflexes.

11. Physiology of airways.

Laboratory research 7

Digestion physiology. Digestion in a mouth and in a stomach

Work 1.Enzymatic properties of saliva in humans.

Objective: Demonstration of substrate specificity of salivary amylase

The object of the study: human

Equipment: multimedia

Stages:

Salivary amylase is mixed with three carbohydrates with different structures. The Trommer`s

reaction for the identification of monosaccharides is performed and the red color which appears

at the end of the reaction proves that only starch is decomposed by the enzyme.

Results:

Table

Resultsof the interaction of saliva with carbohydrates

Type of

carbohydrates

Salivary amylase Temperature The Trommer`s

Reaction

(CuSO4+NaOH)

Starch

Sucrose

Cellulose

12

Conclusion:Salivary amylase is a glycolytic enzyme whose characteristic substrates are

represented by starch and glycogen. It`s activity is enchanced by the Cl- ion and it is the most

efficient at a temperature of 37-38 C and slightly alkaline pH (7.5-8)

Work 2.The influence of pH on the action of pepsin.

Objective: Demonstration of the influence of pH on the efficiency of pepsin

The object of the study: human

Equipment: multimedia

Stages: Incubation of ovalbumin and pepsin at 38C, for 3 hours, with and without hydrochloric

acid, estimating the degree of protein digestion (dimensions of ovalbumin fragments)

Results:

Experiment Temperature Answer

Ovalbumine+pepsin+H2O

Ovalbumine+pepsin+HCl

Ovalbumine+pepsin+HCl+H2O

Conclusion:Pepsin is a proteolytic enzyme which is synthesized as inactive pepsinogen by the

main cells of the gastric glands. Pepsinogen is turned into active pepsin when pH is lower than 5

Work 4.The composition and properties of gastric juice

Purpose: Experimentally study the composition and properties of gastric juice

The object of the study and Equipment: 8 test tubes, thermostat, ice, spirit lamp, tweezers, 2%

pepsin solution in hydrochloric acid (artificial gastric juice), boiled chicken egg protein, pieces

of boiled chicken meat, pieces of raw and boiled beef meat, 10% sodium chloride solution, 2%

solution CuSO4.

Stages and Results:

test

tube

numbe

r

digestive

juice

character

pH protein

character

temp

eratu

re

time digestion results

presence

or

absence

of

insolubl

e

protein

biuret

reaction

1 add 2 ml of

artificial

gastric juice

acidenviron

ment

add boiled

meat

37°C 30-40 min

2 add 2 ml of

artificial

gastric juice

acidenviron

ment

add raw

meat

37°C 30-40 min

3 add 2 ml of

artificial

gastric juice

acidenviron

ment

add

chicken

egg

37°C 30-40 min

4 add 2 ml of

artificial

gastric juice

acidenviron

ment

add

chicken

meat

37°C 30-40 min

5 add 5%

hydrochloric

acid solution

(HCl)

acidenviron

ment

add boiled

chicken

meat

37°C 30-40 min

13

6 add 2 ml of

artificial

gastric juice,

bring to a boil

and cool

acidenviron

ment

add boiled

chicken

meat

37°C 30-40 min

7 add 2 ml of

artificial

gastric juice,

neutralize it

with alkali

alkalineenvi

ronment

add boiled

chicken

meat

37°C 30-40 min

8 add 2 ml of

artificial

gastric juice

acidenviron

ment

add boiled

chicken

meat

0°C 20 min

Conclusion:gastric juice breaks down protein only in an acidic environment at 37 degrees

Laboratory research 8

Digestion physiology.Digestion in intestines.A role of a pancreas and liver in

digestion.Motility of gastrointestinal tract (GIT).Absorption in various departments of a

GIT.

Work 1.The action of bile on fats.

Objective: Demonstrate the emulsification of fats

The object of the study: bile

Equipment:bile, tubes, vegetable oil

Stages:

The first tube pour some oil, add an equal volume of bile. The second tube pour oil and water.

Close the tube and shake a finger.

Results:As a result of agitation in a test tube with bile produced “white milk” - fat emulsion.

Conclusion:Bile emulsifies fats

Work 2.Demonstration of the action of pancreatic lipase in the presence and absence of bile.

Objective: demonstration of the influence of bile on the activity of pancreatic lipase

The object of the study: human

Equipment:multimedia

Stages:

Pancreatic lipase and vegetable oil are introduced in two test tubes with and without bile and

they are kept at a temperature of 37 °C. Then phenolphthalein (a pH indicative which becomes

red when the medium of reaction is alkaline) is added in both test tubes. This proves that

medium is highly acid only in the test tube that contains bile, because fatty acids appear from the

decomposed lipids.

Results:

Experiment Temperature + phenolphtalein

Oil+bile+pancreatic lipase

Oil+bile

Oil+pancreatic lipase

Conclusion:pancreatic lipase is a lipolytic enzyme, which decomposes lipids into glycerol and

fatty acids. The best temperature for the activity of pancreatic lipase is 37-38°C and the best pH

14

is slightly alkaline. The activity of pancreatic lipase is enhanced by bile which has tensioactive

properties, thus enlarging the area of action of this enzyme.

Work 3.The study of the mechanism of absorption of various solutions in the acute experience

(Experience Heidengayn)

Objective: to study the speed of the suction process.

Equipment: operating table, a set of surgical

instruments, syringe, graded burette, polyethylenetube, tray, hypotonic, isotonic solution and

hypertonic solution of sodium chloride, 1% solutionglucose

The object of the study: rat.

Stages: The rat is fixed on the operating table. In the midline of the abdomen is cut in layersskin,

abdominal wall muscles, peritoneum. The section of the small intestine is 10–15 cm long, and

after an incision, cannulas are inserted into both ends of the small intestine and fixed. A

polyethylene tube connected to a graduated burette is put on one cannula, and a polyethylene

tube on the other. Wash the intestine with a syringe and clamp the discharge tube. Fill the system

with tinted isotonic sodium chloride solution and measure the amountsubstance absorbed in 10

minutes due to solution loss in a graded burette. Opening the clamp, remove the non-sucked

solution. The same is done with hypo-and hypertonic solutions. Then, a 1% glucose solution is

introduced into the intestine, and the level of the liquid in the burette is monitored for 10

minutes.

Results:

solution concentration suction time, min amount of solution absorbed

Conclusion: determine which solution is absorbed best

Laboratory research 9

Questionsto the Colloquium

1. Digestion as the main component of functional system on maintenance of level of

nutritious in an organism.

2. Digestion types.

3. Research methods of digestive tract functions.

a) In experiment

a) In clinic

4. Digestive tract innervation.

5. Digestion in a mouth. Structure and properties of salvia.

6. Swallowing.

7. Digestion in a stomach. Composition of gastric juice.

8. Phases of gastric secretion.

9. Structure and properties of pancreatic juice. Regulation of pancreatic secretion.

10. Liver role in digestion.

11. Digestion in a nestis and ileum.

12. Secretion of intestinal juice, its structure, properties, secretion regulation.

13. Digestion in a colon.

14. Motility of a stomach. Evacuation of gastric contents in intestines.

15. Motility of a small and thick intestine.

16. Absorption in various departments of a digestive tract.

15

Laboratory research 10

Allocation physiology. Regulation of kidneyfunctions.

Work 1.Experience Minor

Objective: To learn the method of studying sweating The object of the study: Human Equipment:alcohol, cotton, iodine solution, cotton swabs and hot water, starch

Stages:

The palm should be degreased with alcohol. Apply to the palm of a solution of iodine. When the

solution of iodine dries, apply powdered starch. Remove the starch particles that are not adhered

to the skin. The other hand is immersed in hot water. Watch for changes in the color of the

starch.

Results:Sketch the shape and location of dots and spots on the palms - these are sweat glands.

Conclusion:Treated with starch and iodine solution, the palm becomes blue in contact with

sweat secretions. The intensity of the color varies depending on the degree of sweating activity.

Laboratory research number 11

Physiology of endocrine glands.

Work 1.The effect of insulin and aloxan on blood glucose.

Objective: Demonstration of the effect of insulin on the healthy rat and on the rat with insulin-

dependent diabetes mellitus

The object of the study: rat

Stages:

Collect a blood sample from the normal rat and then another blood sample from the rat with

diabetes mellitus, before and after insulin administration. Measure the blood glucose.

Results:Determine the glucose level (mg/dl)

-normal rat

-aloxan

-insulin

-aloxan+insulin Conclusion: the main metabolic action of insulin is to lower the level of the glucose in the whole

blood

Work 2.Hypoglycemic convulsions in white mice

Objective:show the effect of insulin pancreatic hormone on blood sugar levels and body

condition.

The object of the study: mice

Stages:Each mouse is injected under the skin with 1 Unit of insulin. One mouse is immediately

injected intraperitoneally with 1 ml of 10% glucose solution. Mice are tagged and placed under

separate glass caps to monitor their condition and behavior.

Results: After 40–60 minutes, a mouse that received insulin without glucose shows signs of

hypoglycemic shock (unusual posture, increased breathing, wool swelling, impaired coordination

of movements, clonic convulsions). The higher the temperature at which the mice are kept, the

faster the insulin action. The attack of convulsions can last for about 20 seconds and longer and

lead to the death of the animal. Intraperitoneal administration of 1 ml of 10% glucose solution of

this mouse in a few minutes leads to the restoration of the normal state. In a mouse in which the

glucose solution was injected immediately after insulin injection, seizures and other signs of the

hypoglycemic state are not observed.

Conclusion:The pancreatic hormone - insulin - plays the role of a regulator of carbohydrate

metabolism in the body. An increase in the amount of insulin in the blood leads to a decrease in

the glucose content in it. If, after the introduction of an excessively high dose of insulin, the level

of glucose in the blood decreases to 2.240 mmol/l, then this causes a sharp impairment of brain

16

activity and hypoglycemic shock develops. The only therapeutic measure for hypoglycemic

shock is the introduction of glucose into the body.

Laboratory research number 12

Theme: Metabolism. Thermal control.

Work1.Making diet

Objective: to use formula to estimate metabolic rate at rest, and from that a daily energy

expenditure.

The object of the study: Human

Equipment:the tables of calories (tabl.1,2,3)

Stages:

Meals should be roughly distributed throughout the day. It should start from three meals a day.

For breakfast should account for about 30% of daily calories for lunch - 50%, for dinner - 20%.

Take the daily requirement of calories, proteins, fats and carbohydrates, and divide these values

in the parts corresponding to breakfast, lunch and dinner.

Result:

Food Products Quantity, g The content of

protein, fat,

carbohydrates,

g

Calories,

calories

1. Breakfast

Total:

2. Lunch

Total:

3. Dinner

Total

Total per day:

Conclusion: We compiled a diet, taking into account the ratio of proteins: fats: carbohydrates, calorie intake of

the daily diet

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Table 1

Rates of physiological needs for energy and nutrients for men

Indicators,

( per day)

Group exercise (physical activity ratio) Men

over60

years I (1,4) II (1,6) III (1,9) IV (2,2) V (2,5)

Age groups

18-29 30-39 40-

59 18-29

30-

39 40-59 18-29 30-39 40-59 18-29 30-39

40-

59 18-29 30-39

40-

59

Energy

Energy, kcal 2450 2300 2100 2800 2650 2500 3300 3150 2950 3850 3600 3400 <4200 3950 3750 2300

Protein, g 72 68 65 80 77 72 94 89 84 108 102 96 117 111 104 68

including

animal, g

36 34 32,5 40 38,5 36 47 44,5 42 54 51 48 58,5 55,5 52 34

% of calories 12 12 12 12 12 12 11 11 11 11 11 11 11 11 11 12

Fat, g 81 77 70 93 88 83 110 105 98 128 120 113 154 144 137 77

Fat,% of

calories

30 30 30 30 30 30 30 30 30 30 30 30 33 33 33 30

Carbohydrates,

g

358 335 303 411 387 366 484 462 432 566 528 499 586 550 524 335

18

Table 2

Rates of physiological needs for energy and nutrients for women

Indicators,

( per day)

Group exercise (physical activity ratio) Women over

60 years I (1,4) II (1,6) III (1,9) IV (2,2)

Age groups

18-

29

30-

39

40-

59

18-

29

30-

39

40-

59

18-

29

30-

39

40-

59

18-

29

30-

39 40-59

Energy

Energy, kcal 2000 1900 1800 2200 2150 2100 2600 2550 2500 3050 2950 2850 1975

Protein, g 61 59 58 66 65 63 76 74 72 87 84 82 61

including animal, g 30,5 29,5 29 33 32,5 31,5 38 37 36 43,5 42 41 30,5

% of calories 12 12 12 12 12 12 12 12 12 12 12 12 12

Fat, g 67 63 60 73 72 70 87 85 83 102 98 95 66

Fat,% of calories 30 30 30 30 30 30 30 30 30 30 30 30 30

Carbohydrates, g 289 274 257 318 311 305 378 372 366 462 432 417 284

Table 3

Additional needs for energy and nutrients for women during pregnancy and breast-feeding

Pregnancy (second half) Nursing (1-6 months). Nursing (7-12 months).

Energy

Energy, kcal 350 500 450

Protein, g 30 40 30

including animal, g 20 26 20

Fat, g 12 15 15

Carbohydrates, g 30 40 30

19

20

Laboratory research 13

Questionsto the Colloquium

1. Eliminative organs.

2. Nephron as a structural function unit of a kidney.

3. Forming and composition of primary urine.

4. Tubular reabsorption as the 2nd stage of formation of final urine.

5. Tubular secretion.

6. Composition of final urine.

7. Regulation of activity of kidneys.

a) Humoral regulation;

b) Nervous control;

c) Influence of arterial pressure up on work of kidneys.

8. Act of uresis.

9. Definition, classification of hormones.

10. Interrelation and interaction of endocrine glands.

11. Hormones of an adenohypophysis.

12. Physiology of a thyroid gland.

13. Parathyroid glands.

14. Pancreas physiology.

15. Adrenocortical hormones.

16. Adrenal medulla hormones.

17. Reproductive hormones.

18. Definition of metabolism. Processes of assimilation and dissimilation.

19. Plastic and energetic role of nutrients.

20. Nitrogen equilibrium.

21. Regulation of metabolism.

22. Energy balance of an organism.

23. Basic metabolism. Working metabolism.

24. Hess`s law.

25. Laws of diets.

26. Heat production.

27. Thermal control.

Laboratory research 14

Blood physiology.Physiology of erythrocytes.

Work 1. Determination of the number of erythrocytes in the blood

Objective:To master the method of counting red blood cells using the Goryaev's chamber

Object of research: human blood.

Equipment: microscope, the Goryaev's chamber, mixers, alcohol, a solution of 3% NaCl

Stages:

1. Dilute blood in 200 times with 3% NaCl solution. We collect the blood in the mixer to

the level of 0.5 and bring the solution to the mark of 101 NaCl. Or mix in vitro 4 ml of

3%sodium chloride with 20 μl of blood.

2. Mix the blood with the reagents in the tube. Red blood cells wrinkle and become more

prominent.

3. Prepare the Goryaev's chamber: rub the coverslip to the object glass. The solution is placed in

a chamber and counted red blood cells under a microscope with a small increase in 5 large

squares divided into 16 small ones.

4. The resulting amount of red blood cells inserted in the formula

Results:

21

The calculation of erythrocytes according to the formula:

Number of cells = (B* 4000 *200) / 80 = B * 106/l

Conclusion:We mastered the method of counting erythrocytes in the Goryaev's chamber

Work 2.Determination of hemoglobin (Hb) content in blood by the method of Sahli

Objective:To master the methodology for estimating hemoglobin in blood by the Sahli

colorimetric method and to determine its concentration

Object of research: human blood.

Equipment:Sahlihemoglobinometer, alcohol, cotton, distilled water, and 0.1-normal solution of

hydrochloric acid.

Stages:

1. In a graduated test tube of the hemoglobinometer, pour the acid to the marker.

2. Add 0.02 ml of blood.

3. Shake the tube several times and leave for 7-10 minutes.

4. Add distilled drop of water and mix with a glass rod until the color standard of the hemin.

5. Take the testimony in the lower liquid meniscus.

Results: the result multiplied by 10

Conclusion:The method of Sahli is based on the conversion of hemoglobin into hemin with the

addition of hydrochloric acid to the blood.

Work 3. The calculation of the color index of blood (CIB)

Objective: to evaluate the degree of hemoglobin saturation of each individual red blood cell.

Stages:

1. Using data on the content of erythrocytes and hemoglobin in the examined blood, calculate the

color index of blood by the formula:

Hb (g/l) CIB=---------------------------------------------------------------------------- x3

the number of erythrocytes (the first three digits)

2. Compare the obtained results with the available standards.

Conclusion:In healthy people, the color index of blood ranges from 0.9 to 1.1. If color index less

than 0.9, such a phenomenon is called hypochromasia, more than 1.1 - hyperchromasia.

Work 4. Determination of the erythrocyte sedimentation rate (ESR) by Panchenkov’s

method

Objective: to learn the technique of ESR determination, measure it and give an opinion on the

conformity of the received data to the existing standards.

The object of study: human blood.

Equipment:Panchenkov's device, timer, alcohol, 5% sodium citrate solution.

Stages:

1. Wash the capillary with sodium citrate. Collect the citrate to the label of reagent P and blow to

the watch glass.

2. Draw blood into the capillary to the K mark twice and blow it out onto the watch glass. Thus,

we mix sodium citrate and blood in a ratio of 1: 4.

3. Fill the capillary with a mixture to the mark 0, put in a tripod for 60 minutes.

Results:Estimate the height of the plasma column in mm after 1 hour

Conclusion:ESR size normally ranges from 4 to 10 mm/hr

Work 5.Blood sampling from the finger Objective:

22

The object of study: human

Equipment: alcohol, cotton, sterile scarifier

Stages: Wash your hands with soap and running water, and dry them. Make blood collection

from the thumb or ring finger of your left hand (it is permissible to receive blood from any other

finger). A blood taker must wear rubber gloves. Wipe the skin of the fingertip with a cotton swab

moistened with 70% alcohol and wait for it to dry. Use your left hand to slightly squeeze the

pulp of the finger in the area of the intended injection. Take a sterile scarifier in your right hand,

orienting it strictly perpendicular to the skin surface at the injection site. The most convenient

place to puncture the skin is the point to the left of the midline at some distance from the nail.

Make a prick to the entire depth of the needle point, while cutting through the skin across the

fingerprint lines. Remove the first drop of blood because it contains inadvertent impurities,

lymph and damaged corpuscles. Next, take the blood for the necessary tests. After the end of the

blood sampling, apply a cotton swab moistened with alcohol or iodine solution to the puncture

site. Finger blood draw in special in each case glass capillaries to standardize the process of

blood collection.

Conclusion: The correct receipt of capillary blood is one of the decisive conditions that ensure

the accuracy and reproducibility of the results. The total time spent on blood collection should

not exceed 2 - 3 minutes. In the taken blood there should be no signs of clotting.

Laboratory research 15

Physiology of leukocytes and thrombocytes. Hemolysis

Work 1. Determination of the number of leukocytes in the blood

Objective: To master the method of counting white blood cells using a counting chamber to

determine their blood and give an opinion on the conformity of the data to the existing standards.

Object of research: human blood.

Equipment: microscope, the Goryaev's chamber, mixers, alcohol, watch glass, a solution of 5%

acetic acid with methylene blue (Türk solution) stored blood.

Stages:

1. To dilute the blood in 20 times with a solution of the Turk. Collect blood into the mixer to the

level 1 and get the Türk solution up to level 11.

2. Mix the blood with the reagent in the watch glass. The solution of acetic acid hemolysis of

erythrocytes. Methylene blue stains the nuclei of leukocytes.

3. Prepare the Goryaev's chamber. The solution is placed in the chamber and counted white

blood cells under a microscope with a small increase in 25 large squares of 16 small separated

ones.

4. Substitute the number in the formula

Results:

The calculation of leukocytes according to the formula:

Number of cells = (B* 4000 *20) / 400 = B * 106/l

Conclusion:We mastered the method of counting leukocytes in the Goryaev's chamber

Work 2.Determination of blood viscosity

Equipment: viscometer

Stages: The determination of blood viscosity is based on a comparison of the speed of blood and

distilled water in the same capillaries under vacuum at room temperature. The determination is

carried out in the instrument viscometer. In the right capillary pipette of the viscometer dial

distilled water to the mark "0". In the left capillary, blood is sucked from the finger to the zero

23

mark. Turn the three-way valve in such a way as to connect both capillary pipettes with a rubber

tube through which air is drawn from both pipettes to form a vacuum.

Results.The columns of water and blood move forward at different speeds, which depend on

viscosity. As soon as the blood column reaches the “1” mark, the air intake is stopped. During

this time, water, which has a lower viscosity, moves much further than blood. Blood viscosity is

determined by the length of the path traveled by water, which is measured on a scale of a

graduated pipette. Blood viscosity is normal for men 4.3–5.4, and for women 3.9–4.9 scale

points.

Conclusion.There is a dependence of blood viscosity on the number and volume of erythrocytes,

the total protein content and the ratio of its fractions in plasma, as well as the blood carbon

dioxide content. An increase in viscosity is observed in case of thickening of the blood and some

types of leukemia (erythremia, myelofibrosis), a decrease in case of anemia.

Laboratory research 16

Blood fibrillation. The doctrine about blood groups

Work 1.Determination of blood groups by Colyclons.

Objective: Determine blood group ABO system reagents Colyclons

Equipment:Colyclons in bottles: anti-A - pink color, anti-B - blue, anti-AB - colorless, tablet,

glass rod, cotton, alcohol, stored blood.

Stages:

1. Applied to plate or plate individual pipettes Colyclons anti-A, anti-B and anti-AB one large

drop (0.1 ml) under appropriate labels.

2. Near drops antibody coated on one small drop of blood examined (0.01 - 0.03 ml).

3. Mix the blood with a reagent shaking plate.

4. Tracking the progress of the reaction with Colyclons visually under gentle shaking the plate

for 3 minutes. Agglutination of red blood cells with Colyclons usually occurs in the first 3 - 5

seconds, but monitoring should be carried out 3 minutes later due to the appearance of the

agglutination with erythrocytes containing weak variety of antigens A or B.

5. The result of the reaction in each drop can be positive or negative. A positive result is

expressed as the agglutination (gluing) of red blood cells. Agglutinates appear as small red

assembly rapidly mixing in large flakes. When a drop backlash is uniformly reddened,

24

agglutinates it can not be detected.

Interpretation of the results of agglutination reaction is shown in Table Colyclons

Results:

The presence of agglutination indicates the presence of blood groups.

the study of

blood group

Standard erythrocytes

anti-А anti-В anti-АВ

0(I) _ _ -

А(II) + - +

В(III) - + +

АВ(IV) + + +

Note: The plus sign (+) denotes the presence of agglutination, the sign (-) - No

Conclusion:Determination of blood groups by Colyclons

Work 2.Determination of Rh by Colyclons.

Objective: Determination of Rh reagents Colyclons

Equipment:Colyclons in bottles: anti-D, tablet, glass rod, cotton, alcohol, stored blood.

Stages:

1. Applied to plate or plate individual pipettes Colyclonsanti-D one large drop (0.1 ml) under

appropriate labels.

2. Near drops antibody coated on one small drop of blood examined (0.01 - 0.03 ml).

3. Mix the blood with a reagent shaking plate.

4. Tracking the progress of the reaction with Colyclons visually under gentle shaking the plate

for 3 minutes. Agglutination of red blood cells with Colyclons usually occurs in the first 3 - 5

seconds, but monitoring should be carried out 3 minutes later due to the appearance of the

agglutination with erythrocytes containing weak variety of antigens D.

5. The result of the reaction in each drop can be positive or negative. A positive result is

expressed as the agglutination (gluing) of red blood cells. Agglutinates appear as small red

assembly rapidly mixing in large flakes. When a drop backlash is uniformly reddened,

agglutinates it can not be detected.

Results:

The presence of agglutination indicates the presence of Rh-factor. When agglutination with

Colyclon anti-D appears, the blood is Rh-positive, in the absence of agglutination, the blood is Rh-

negative

25

Laboratory research 17

Questionsto the Colloquium

1. Composition of blood.

2. Blood functions.

3. Physiological constants of blood.

4. Blood plasma. Structure, osmotic and oncotic pressure.

5. Erythrocyte. Structure and functions.

6. Regulation of erythropoiesis.

7. Hemoglobin and its combinations.

8. Erythrocyte sedimentation rate (ESR). Mechanism of ESR.

9. Leukocytes, quantity, leukogram.

10. Regulation of granulocytopoiesis and monocytopoiesis.

11. Lymphocytes. Regulation of lymphocytopoiesis.

12. Thrombocytes, quantity, functions.

13. Types and mechanism of hemolysis.

14. Blood fibrillation process. Blood fibrillation factors.

15. Blood fibrillation phases.

16. The anticoagulation system

17. The doctrine about blood groups. Rh factor.

Laboratory research 18

Practical skills

1. Determination of the number of erythrocytes in the blood

2. Determination of hemoglobin content in blood by the method of Sahli

3. The calculation of the color index of blood

4. Determination of the erythrocyte sedimentation rate by Panchenkov`s method

5. Determination of the number of leukocytes in the blood

6. Observation of different types of hemolysis

7. Determination of osmotic resistance

8. Determination of blood groups

9. Determination of Rh

10. Determination of coagulation time

11. Determination of bleeding time by Duke

12. Listening to heart tones.

13. Definition of blood pressure by the method of Korotkov.

14. The ECG recording.

15. Palpation of the pulse.

16. Measurement of the vital capacity of the lungs and its components.

17. Pneumography.

18. Calculation of basal metabolic rate.

19. The study of the reaction time.

20. Dynamometry.

21. Assessment of the field of view.

22. Assessment of visual acuity.

23. Research methods of taste sensitivity.

Literature

1. SándorBorbély, LászlóDétári, TündeHajnik, KatalinSchlett, Krisztián,Tárnok

Attila Tóth, Petra Varró, IldikóVilági. Physiology practical. Budapest, 2013 – 220 p.

2. Guide to practical exercises in normal physiology [Text]: textbook / ed. CM. Budylina,

V.M. Smirnova. - 3rd ed., Erased. - M.: Academy, 2010 .-- 336 p.