Tài liệu Nghiên cứu sự biến đổi một số chỉ số sinh học và tác dụng của điều trị tái tăng áp trong bệnh giảm áp cấp tính thực nghiệm.

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.