XIA Qiang（夏强） , MD & PhD

Email: xiaqiang@zju.edu.cn

Room 518, Block C, Research Building

Zhejiang University School of Medicine

Introduction toExperimental Physiology

Course Head: Dr. Xia Q

Instructors: Dr. Wang HP Dr. Yu YQ Dr. Zhang X

Assistants: Miss Li Yuying, Mr Pan Libiao, Miss Zhou Rui

First assignment: 10 groups, group leader

Group on duty: Rotation: one group/week

Aim of Experimental Physiology

1. Develop students’ operational skills and ability to write

experimental reports

2. Develop students’ scientific thinking and ability to ask

questions, to analyze problems and to solve problems

3. Develop students’ attitude to practical work, normal

operational methods, and cooperation based on team

At the end of the course, students should be able to:

1. Understand the measurement of basic physiological indices

2. Perform physiological experiments and possess the essential surgical

techniques required

3. Understand the essential techniques of electrophysiology

4. Analyze experimental results and write experimental reports independently

5. Understand the general principles and methods of design of physiological

experiments

Aim of Experimental Physiology

Lab requirements

1. White coat

2. Preview

3. Following the lab manual and the regulation

4. Review

5. Written report

6. Group member contribution

Experimental Animals

Toad Rat or mouse Rabbit

Scalpel and blade

Instruments

Scissors

Instruments

Hemostat or Clamp

Instruments

Forceps

Other instruments will be introduced as needed Frog surgical instruments

Mammal surgical instruments

Instruments

Transducers

A transducer is an electronic device that converts energy from one form to another

Pressure transducer

Force transducer (Displacement transducer)

Force transducer provides an output proportional to the force applied.

A pressure transducer converts pressure into an analog electrical signal. Although there are various types of pressure transducers, one of the most common is strain-gage based transducer.

Data Acquisition System

The RM6240 multi-channel physiological recording and processing system will be used

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Working window All essential controls for recording data are provided within the Chart

Application window.

Tool Bar The Tool Bar consists of a row of buttons which provide shortcuts for common

tasks. The button functions are displayed if the pointer is held over them.

Data Measurement: After you stop recording, you can use these functions to measure the data and

get values. All these values can be output to an EXCEL file.

Hidden function buttons

Hidden function buttons

Hidden function buttons

Hidden function buttons

Hidden function buttons

Experiment Ready-made experimental schemes for physiological

experiments

• The correct parameters have been preset.

To use the functions in the “create experiment” menu to build a new scheme for your own experiment.

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Writing Experimental Reports

(1) General requirements Use the required report format. Complete each item and mark clearly your name and number. Record the date of the experiment. Leave a space for the signature of the teacher. Please stick to the standard format, provide accurate charts, use legible handwriting or type, and submit the report on time. No plagiarism.

(2) Basic format of the report Some physiological experiments emphasize surgical methods such as

the nerve-muscle preparation; others emphasize measurement of a phenomenon such as gastrointestinal motility; while some lay particular emphasis on the results and analyses, such as the influence of factors on urine formation. Most experiments include all three dimensions.

The general experimental report format: Name: Number: Class: Year: Date: Score:

Title

1. Objectives

2. Principle

3. Animal

4. Apparatus (list)

5. Methods and Procedures (outline)

6. Results (detailed)

7. Discussion (detailed)

8. Conclusion

Watch the video of animal experimental techniques http://10.71.149.149/

Course website http://m-learning.zju.edu.cn

Practice one by one. Every one should be sure to know how to use it

Preview the experiment in the lab manual for next week

Case: Hereditary Spherocytosis

A 20-year-old woman suffers from anemia and occasional jaundice. A thorough review of her medical records reveals that over the past l0 years she has had episodes of more severe anemia, usually after periods of febrile illness. The patient has a markedly enlarged spleen. Microscopic examination of the patient's blood showed a large number of microspherocytes (red blood cells [RBCs] that are round and somewhat smaller than erythrocytes). The osmotic fragility (measured by putting RBCs in hypotonic solutions) was much greater than that of RBCs from healthy individuals. When the patient's erythrocytes were incubated in a buffer solution at 37C under sterile conditions, the fraction of the RBCs that were hemolyzed was much larger than the hemolyzed fraction from a healthy individual. This "autohemolysis" could be greatly diminished by including glucose and adenosine triphosphate (ATP) in the RBC incubation solution. RBCs from fresh blood had a normal content of Na+ and K+. The permeabilities of the patient's erythrocyte membranes to Na+ and K+ were found to be about three times normal. The level of Na+, K+-ATPase in the patient's RBC membranes was also about three times the level in RBCs from healthy individuals. The average life span of the patient's erythrocytes was well below the normal life span. When an aliquot of the patient's RBCs was labeled and injected intravenously into a healthy individual, the patient's RBCs had a markedly reduced survival time compared with normal RBCs. When labeled RBCs from a healthy individual were infused into the patient, the survival time of the normal RBCs was comparable with their survival time in the donor. The patient's spleen was removed, and after the splenectomy, the patient's anemia was largely ameliorated.

Questions

1. Why should the patient's erythrocytes have a greater osmotic fragility than RBCs from healthy individuals?

2. Why might the patient's RBCs "autohemolyze" more rapidly than normal erythrocytes when they are incubated at 37C under sterile conditions?

3. Why should including glucose and ATP in the incubation mixture diminish the extent of autohemolysis?

4. Why should the patient's RBCs have a reduced life span? What might the spleen have to do with this?

5. What is proved by the observations that the patient's RBCs have a reduced life span in the circulation of a healthy individual and that the RBCs of a healthy individual have a normal life span in the patient's circulation?

6. Why might the patient have more severe episodes of anemia following febrile illnesses?

7. Why should splenectomy largely correct the patient's anemia?

Thank you for your attention!