MINISTRY OF EDUCATION AND TRAINING MINISTRY OF DEFENCE
VIETNAM MILITARY MEDICAL UNIVERSITY
CAO HONG PHUC
RESEARCH ON SOME OF BIOLOGICAL CHANGES AND
EFFECTS OF RECOMPRESSION TREAMENT IN
EXPERIMENTAL DECOMPRESSION SICKNESS
MEDICAL DORTOR PHILOSOPHY THESIS
HA NOI-2018
MINISTRY OF EDUCATION AND TRAINING MINISTRY OF DEFENCE
VIETNAM MILITARY MEDICAL UNIVERSITY
CAO HONG PHUC
RESEARCH ON SOME OF BIOLOGICAL CHANGES AND
EFFECTS OF RECOMPRESSION TREAMENT IN
EXPERIMENTAL DECOMPRESSION SICKNESS
Branch: Bio - Medicine Sciense
Code: 972 01 01
MEDICAL DORTOR PHILOSOPHY THESIS
SUPERVISORS:
1. Assoc.Prof. Nguyen Tung Linh, PhD
2. Assoc.Prof. Dang Quoc Bao, PhD
HA NOI-2018
i
COMMITMENT
I declare that this is my onw thesis which was carried out by scientific
supports of superviors.
All results that were presented in this thesis have been authentic and
published in scientific articles. The thesis is not repeated to previous thesises
or projects. If there is something wrong, I will take my responsibility.
Author
Cao Hong Phuc
ii
ACKNOWLEGEMEND
After years of in-depth research, this thesis has been completed. I would
like to show my sincerely thanks to the Party Committee, Broad of Directors:
Vietnam Military Medical University, Military Hospital No 103, National
Institute Of Hygiene And Epidemiology for their facilitation.
I would also like to thank: Department of Medicine of Arms, Department
of Medical Physiology, Department of Experimental Operation, Department of
Medical Physics (Vietnam Military Medical University), Departemt of
Hematology and Tranfusion, Patho-Histology and Judical Medicine (Military
Medical No 103), Laboratory of Micro-Structure (Virus Department, National
Institute Of Hygiene And Epidemiology), Department of Post - Graduate
Training for their technical help in my study.
In addition, I am completely grateful to my suppervisors: Asscoc.Prof
Nguyen Tung Linh, PhD (first suppervios), Asscoc.Prof Dang Quoc Bao, PhD
(second suppervios). The doors of their offices were always open whenever I
ran into a trouble spot about my reseach. They consistently allowed this paper
to be my own work, steered me in the right direction in experimental period.
They also set good examples for me to follow in doing all scientific tasks.
Besides, I would like to express my heartfelt gratitude to my parents and
parents - in - law for their bringing up and supporting me to make this work
possible. Last but not least, I am deeply indebted to my wife and my son for
being on my side, encouraging me a lots throughout my years of study.
AUTHOR
Cao Hong Phuc
iii
CONTENT
SUBTITLE PAGE
COMMITMENT ................................................................................................ i
ACKNOWLEGEMEND ................................................................................... ii
CONTENT ....................................................................................................... iii
ABBREVIATIONS AND DEFINITIONS .................................................... viii
LIST OF TABLES .......................................................................................... xii
LIST OF FIGURES........................................................................................ xvi
INTRODUTION ............................................................................................... 1
CHAPTER 1. OVERVIEW .............................................................................. 3
1.1. CLINICAL, INVESTIGATING CHARACTERISTICS OF
ACUTE DECOMPRESSION SICKNESS ..................................... 3
1.1.1. Causes and risk factors ............................................................... 3
1.1.2. Clinical characteristics ................................................................ 7
1.1.3. Investigating characteristics ...................................................... 12
1.1.4. Diagnosis and classification...................................................... 14
1.1.5. Treatment .................................................................................. 16
1.2. INFLUENCES OF THE BUBBLES IN DECOMPRESSION
SICKNESS ....................................................................................... 20
1.2.1. Bubble formation in the decompression sickness ..................... 21
1.2.2. Influences of the bubbles in decompression sickness .............. 22
1.3. STUDIES ON ANIMAL MODELS AND BIOLOGICAL
CHANGES IN ACUTE DECOMPRESSION SICKNESS ......... 24
iv
1.3.1. Studies on animal models in decompression sickness.............. 24
1.3.2. Some studies of peripheral blood cell changes in acute
decompression sickness .......................................................... 26
1.3.3. Some studies of coagulation changes in decompression
sickness ................................................................................... 28
1.3.4. Some studies of biochemical changes in acute decompression
sickness.................................................................................... 30
CHAPTER 2. SUBJECTS AND METHODS ................................................ 34
2.1. SUBJIECTS ...................................................................................... 34
2.1.1. Subjects ..................................................................................... 34
2.1.2. Laboratories .............................................................................. 34
2.1.3. Period ........................................................................................ 35
2.2. METHODS ....................................................................................... 35
2.2.1. Study design .............................................................................. 35
2.2.2. Study Issues .............................................................................. 40
2.2.3. Study equipments ...................................................................... 40
2.2.4. Research indexes and methods ................................................. 42
2.2.5. Data Analysis ............................................................................ 53
CHAPTER 3. RESULTS ................................................................................ 55
3.1. ETABLISHING THE EXPERIMENTAL ACUTE
DECOMPRESSION SICKNESS MODEL .................................. 55
3.1.1. Influence of some diving factors on acute decompression
sickness ................................................................................... 55
v
3.1.2. Comparision the efficacy of models to cause the
decompression sickness .......................................................... 60
3.2. CHANGES IN HEMATOLOGICAL, BIOCHEMICAL AND
PATHOHISTOLOGICAL INDEXES IN ACUTE
DECOMPRESSION SICKNESS.......................................................... 62
3.2.1. Changes in hematological indexes in acute decompression
sickness rabbits ....................................................................... 62
3.2.2. Changes in some of biochemical indexes in acute
decompression sickness rabbits .............................................. 72
3.2.4. Some of pathohistological characteristics in acute
decompression sickness rabbits .............................................. 73
3.3. EFFECTS OF RECOMPRESSION TREATMENT IN
ACUTE DECOMPRESSION SICKNESS ................................... 83
3.3.1. Effects on clinical symptoms in acute decompression
sickness rabbits ....................................................................... 83
3.3.2. Effects on hematological index in acute decompression
sickness rabbits ........................................................................ 85
3.3.3. Effects of recompression treatment on some biochemical
indexes in in acute decompression sickness rabbits ............... 94
CHAPTER 4. DISCUSSION .......................................................................... 97
4.1. ANIMAL MODEL OF EXPERIMENTAL ACUTE
DECOMPRESSION SICKNESS ............................................... 97
4.1.1. Influence of some diving factors on experimental acute
decompression sickness .......................................................... 97
vi
4.1.2. Effecting comparison of experimental acute
decompression sickness of models .................................... 101
4.2. CHANGES IN SOME HEMATOLOGICAL,
BIOCHEMICAL, AND PATHOHISTOLOGICAL
INDEXES IN EXPERIMENTAL ACUTE
DECOMPRESSION SICKNESS ................................................ 104
4.2.1. Changes of some hematological indexes in experimental
acute decompression sickness rabbits................................... 104
4.2.2. Platelet activation in experimental acute decompression
sickness ................................................................................. 114
4.2.3. Changes of some biochemical indexes in acute
decompression sickness rabbits ............................................ 118
4.2.4. Changes of pathohistological indexes in experimental
acute decompression sickness rabbit .................................... 120
4.3. EFFECTS OF RECOMPRESSION TREATMENT ON
EXPERIMENTAL ACUTE DECOMPRESSION
SICKNESS ..................................................................................... 123
4.3.1. Effects of recompression treatment on clinical symptoms
in experimental acute decompression sickness rabbits ........ 123
4.3.2. Effects of the recompression treatment on haematological
indexes in acute decompression sickness rabbits ................. 128
4.3.3. Effect of recompression treatment on biochemical indices
in acute decompression sickness rabbit ................................ 134
CONCLUSION ............................................................................................. 137
vii
SUGGESSTIONS ......................................................................................... 139
LIST OF PUBLISHED ARTICLES RELATED TO
RESULTS OF THESIS ................................................................................. 140
REFFERENCES............................................................................................ 141
APPENDIXES .............................................................................................. 158
viii
ABBREVIATIONS AND DEFINITIONS
Num
Abbre
Deffinition
1. 2D
Two Dimension
2. ADP
Adenosine diphosphate
3. ALT
Alanine transaminase
Vietnamese Meaning
Hai chiều
Enzym chuyển amin
Alanine
4. APTT
Activated Partial
Thời gian thromboplastin
Thromboplastin Time
hoạt hóa từng phần
5. Ar
Argon
Khí argon
6. ARN
Acid Ribonucleic
A-xít Ribonucleic
7. AST
Aspartate transaminase
Enzym chuyển amin
Aspartate
8. ATA
Atmospheres Absolute
9. ATP
Adenosine triphosphate
10. Aβ
β-amyloid
11. BTG
Beta thromboglobulin
12. Conc
Concentration
13. CPK
Creatine phosphokinase
14. CSF
Cerebrospinal fluid T-Tau
T-tau
15. CT
Áp suất khí quyển tuyệt đối
Nồng độ
Protein T-Tau trong dịch
não tủy
Computerized
Chụp cắt lớp vi tính
Tomography
16. DCS
Decompression sickness
Bệnh giảm áp
ix
Num
Abbre
17. DWI
Deffinition
Vietnamese Meaning
Diffusion Weighted
Imaging
18. EDTA
Ethylene diamine tetra
Chất chống đông EDTA
acetic acid
19. ET-1
Endothelin 1
20. Fib
Fibrinogen
21. FSF
Fibrin Stabilizing Factor
Yếu tố ổn định Fibrin
22. G
Group
Nhóm
23. GFAP
Glial fibrillary acidic
protein
24. Hb
Hemoglobin
25. HCl
Hydrochloric acid
26. Hct
Hematocrit
27. HE
Hematoxylin-Eosin
A-xit Clohydric
Thuốc nhuộm
Hematoxylin-Eosin
28. HMWK
High Molecular Weight
Kininogen khối lượng
Kininogen
phân tử cao
29. HRP
Horseradish peroxidase
30. HSP 70
Heat Shock Protein 70
Protein sốc nhiệt 70
31. ICAM-1
Intercellular Cell Adhesion
Chất liên kết tế bào 1
Mollecular - 1
32. IL-1β
Interleukin 1 beta
33. IL-6
Interleukin 6
34. KLPT
Khối lượng phân tử
x
Num
Abbre
Deffinition
Vietnamese Meaning
35. LDH
Lactate dehydrogenase
36. MCH
Mean corpuscular
Hàm lượng hemoglobin
hemoglobin
trung bình hồng cầu
Mean corpuscular
Nồng độ hemoglobin
hemoglobin concentration
trung bình hồng cầu
Mean corpuscular volume
Thể tích trung bình hồng
37. MCHC
38. MCV
cầu
39. MDA
Malodialdehyde
40. MH
Mô hình
41. MPV
Mean platelet volume
Thể tích trung bình tiểu
cầu
42. NSE
Neuron-Specific Enolase
43. OR
Odd Ratio
Tỷ suất chênh
44. Para
Paralysis
Liệt
45. PF4
Platelet Factor 4
Yếu tố 4 tiểu cầu
46. Post
Post-experiment
Sau gây bệnh
47. Pre
Pre-experiment
Trước gây bệnh
48. PT
Prothrombin Time
Thời gian prothrombin
49. RR
Relative risk
Nguy cơ tương đối
50. sGPV
Soluble Glycoprotein V
Glycoprotein V hòa tan
51. T2WI
T2 Weighted Image
52. TAT
Thrombin-Antithrombin
Phức hợp thrombin -
Complex
antithrombin
Tumor necrosis factor
Yếu tố hoại tử u alpha
53. TNF-α
alpha
xi
Num
Abbre
Deffinition
54. TT
Thrombin Time
55. TTR
Transthyretin
Vietnamese Meaning
Thời gian thrombin
xii
LIST OF TABLES
Num
2.1.
Table
Page
Classification of clinical severity in experimental acute
decompression sickness rabbits .................................................. 44
2.2.
Paralysis classification in acute decompression sickness rabbits ........ 44
2.3.
Classification of post-treatment response ................................... 44
2.4.
Bubble grade in experimental decompression sickness .............. 52
3.1.
The rabbit number of decompression sickness with variable
bottom time ................................................................................. 55
3.2.
Clinical severity of rabbits after experiment .............................. 56
3.3.
The number of death rabbit after experiment with variable
bottom time ................................................................................. 56
3.4.
The rabbit number of decompression sickness with variable
ascent rate.................................................................................... 57
3.5.
Clinical severity of decompression sickness ............................. 57
3.6.
The number of death rabbits after decompression ...................... 58
3.7.
The rabbit number of decompression sickness .......................... 58
3.8.
Clinical severity and consecutive compression ......................... 59
3.9.
The number of death rabbit and consecutive compression ........ 59
3.10.
Influence of some diving factors on decompression sickness ... 60
3.11.
The rate of decompression sickness and survival of rabbits
after compressing as four models ............................................... 61
3.12.
Clinical severity of rabbit after compressing as four models .... 61
3.13.
Bubble grade of rabbit after compressing as four models ......... 62
xiii
Num
3.14.
Table
Page
Changes in red blood cell count according to clinical
severity in acute decompression sickness .................................. 62
3.15.
Changes in hemoglobin concentration according to
clinical severity in acute decompression sickness ..................... 63
3.16.
Changes in hematocrit according to clinical severity
in acute decompression sickness ................................................ 63
3.17.
Changes in MCV, MCH, MCHC according to clinical
severity in acute decompression sickness .................................. 64
3.18.
Changes in white blood cell count according to clinical
severity in acute decompression sickness .................................. 65
3.19.
Changes in white blood cell percentage according to
clinical severity in acute decompression sickness ..................... 65
3.20.
Changes in platelet count according to clinical severity
in acute decompression sickness ................................................ 66
3.21.
Change in MPV according to clinical severity in acute
decompression sickness .............................................................. 67
3.22.
PT change according to clinical severity .................................... 68
3.23.
APTT change according to clinical severity ............................... 69
3.24.
Plasma fibrinogen concentration according to clinical
severity in acute decompression sickness .................................. 71
3.25.
Changes in plasma platelet markers between pre and
post acute decompression sickness ............................................. 71
3.26.
Changes in liver enzyme activity ................................................ 72
3.27.
Changes in plasma concentration of ure and creatinin ............... 73
3.28.
The rabbit number of bubbles in micro pathohistological
images after acute decompression sickness in 1st phase ............. 78
xiv
Num
3.29.
Table
Page
Distribution of rabbits according to bubble grade and
paralysis grade in acute decompression sickness rabbits .......... 80
3.30.
Rabbit number of platelet activation after acute
decompression sickness .............................................................. 83
3.31.
Changes in clinical symptoms between pre and post
treatment in acute decompression sickness rabbits .................... 83
3.32.
Changes in clinical severity between pre and post treatment
in acute decompression sickness rabbits..................................... 84
3.33.
Treatment responding rabbits according to clinical category ..... 84
3.34.
Treatment responding rabbits according to clinical severity ...... 85
3.35.
Effects of recompression treatment on red blood
count in acute decompression sickness rabbits........................... 85
3.36.
Effects of recompression treatment on hemoglobin
concentration in acute decompression sickness rabbits .............. 86
3.37.
Effects of recompression treatment on hematocrit in acute
decompression sickness rabbits .................................................. 87
3.38.
Effects of recompression treatment on MCV, MCH
and MCHC in acute decompression sickness rabbits ................. 87
3.39.
Effects of recompression treatment on white blood
count in acute decompression sickness rabbits........................... 88
3.40.
Effects of recompression treatment on leukocyte
percentage in acute decompression sickness rabbits .................. 89
3.41.
Effects of recompression treatment on platelet
count in acute decompression sickness rabbits........................... 91
3.42.
Effects of recompression treatment on MPV in acute
decompression sickness rabbits .................................................. 91
xv
Num
3.43.
Table
Page
Effects of recompression treatment on PT in acute
decompression sickness rabbits .................................................. 92
3.44.
Effects of recompression treatment on APPT in acute
decompression sickness rabbits .................................................. 93
3.45.
Effects of recompression treatment on plasma
concentration of fibrinogen in acute decompression
sickness rabbits ........................................................................... 93
3.46.
Effects of recompression treatment on liver enzyme
activities in acute decompression sickness rabbits ..................... 94
3.47.
Effects of recompression treatment on plasma ure
and creatinin concentration in acute decompression
sickness rabbits ........................................................................... 95
xvi
LIST OF FIGURES
Num
Figure
Page
1.1.
Cutis mamorata ............................................................................ 9
1.2.
Nitrogen bubbles in the left atrial chamber .............................. 13
1.3.
Nitrogen bubble in spinal cord of decompression sickness dog .. 14
1.4.
Liver injury caused by the bubbles ........................................... 23
1.5.
Platelet ultrastructure ................................................................ 27
2.1.
Diagram of 1st phase of study. .................................................. 36
2.2.
Diagram of 2st phase of study ................................................... 37
2.3.
Recompression treatment protocol .......................................... 39
2.4.
Experimental recompression chamber ..................................... 41
2.5.
Gold coating on electromicroscope specimen .......................... 53
3.1.
The relation between platelet count and survival time
post experiment ........................................................................... 66
3.2.
Change in platelet count between pre and post
decompression sickness according to paralysis degree .............. 67
3.3.
Relation between PT and survival of rabbits after acute
decompression sickness .............................................................. 68
3.4.
Relation between PT and platelet count...................................... 69
3.5.
Relation between APPT and survival time of rabbits ................ 70
3.6.
Relation between APTT and platelet count in rabbits
after acute decompression sickness ............................................ 70
3.7.
Vessels in normal and acute decompression sickness rabbit ....... 74
3.8.
Adominal muscle vessels in normal and acute
decompression sickness rabbit .................................................... 74
3.9.
Lumbar branch of vena cava in normal and acute
decompression sickness rabbit .................................................... 75
xvii
Num
3.10.
Figure
Page
Mesenteric veins in normal and acute decompression
sickness rabbit ............................................................................. 75
3.11.
Vena cava in normal and acute decompression .......................... 76
3.12.
Abdomincal aorta in normal and acute decompression .............. 76
3.13.
Bubbles in the blood drained out from the incised organs ......... 77
3.14.
Abdominal muscle in acute decompression sickness rabbits ...... 77
3.15.
Vein of abdominal muscle in acute decompression sickness
rabbits (HE x100)........................................................................ 77
3.16.
Liver tissue in in acute decompression sickness rabbits ............. 78
3.17.
Lung tissue in acute decompression sickness rabbits ................. 78
3.18.
Kidney cappilary in acute decompression sickness rabbits ........ 78
3.19.
Kidney tissue in acute decompression sickness rabbits .............. 78
3.20.
Inside heart in acute decompression sickness rabbits ................. 78
3.21.
Coronary vessles in acute decompression sickness rabbits ........ 78
3.22.
Distribution of clinical severity according to bubble grade
in acute decompression sickness rabbits .................................. 80
3.23.
Survival time of rabbits and bubble grade in acute
decompression sickness rabbits .................................................. 81
3.24.
Platele shape in normal rabbits .................................................. 82
3.25.
Pseudopods and oblong shape of platelet in acute
decompression sickness rabbits .................................................. 82
3.26.
Platelet deformation and degeneration in acute
decompression sickness rabbits .................................................. 82
1
INTRODUTION
Decompression sickness is a occupational disease of divers, causes
noticeable economic harmfulness, a lot of complication and may cause death.
Although patients were treated completely, some worse patients could still
bare seriously outcomes [1].
The number of decompression sickness patients has been a not - small
number in occupational disease group. According to British Diving Coucil,
the death patients were 98 cases in 2010 [2]. In Denmark, Svendsen J.C. et al
realised that there were 205 decompression sickness patients from 1993 to
2013, average 14 patients/year [3]. In Vietnam, Nguyen Van Non et al
reported that the rate of decompression sickness was 22% of fishery
population in Northern Bay [4]. It is clear that decompression sickness should
be put in high consideration.
So far, most reseachers presumed that bubble is a mainly factor causing
disorders of this desease [5], [6]. Looking at ultrasound images, bubbles were
seen at vein, heart chambers, arteriole, thigh artery [6], [7]. When applied
recompression treatment to minimise the bubbles, it was seen that clinical
symtomes were improved [8]. Therefore they played an importanted role in
machenism of decompession sickness. The relation, however, between
bubbles and risk, severity and prognosis of this desease has not been studied
in detail.
In reality, some patients were applied by recompression treatment did
not recover completely [9], [10]. Moreover, a small number of patients got
mild improvement by using antiplatetet drugs (the drugs could not decrease
bubble grade ) [11]. Hence, a part form bubble role, there were other
biological disorders which contributed to the disease machenism.
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