Study Of Forensic Pathologic Diagnosis In Anaphylactic Shock And Sudden Death From Hypersensitivity

Investigation of epidemiological indicated that incidence of the hypersensitivity which is 21 per 100,000 person is increasing with the development of industrialization and the hypersensitivity disease have become the global disease that threated people health acutely.It is common that anaphylactic shock or sudden death result from allergy in clinical medicine and forensic medicine.Every year there are approximate 154 fatal anaphylacticus episodes during 1,000,000 hospitalzed subjects internationally and death rate of anaphylactic shock in the United States is 1%to 3%according to references.The sudden deaths from other causes are possiblebly suspended as the sudden death from hypersensitivity in medicolegal expertise. Therefore identification of anaphylactic shock and sudden death from hypersensitivity is very significant in medicolegal expertise.The medicolegal expertise involving in sudden death from hypersensitivity is about 10%in the medicolegal expertise of medical tangle at resent years according to our laboratory data.Hypersensitivity disease is the study hot spot.Although scholars have researched much on pathogenesy,clinical diagnosis and clinical therapy of hypersensitivity disease,we still can not find the specialpathological changes on autopsy and histological examination in the dead body sudden death from hypersensitivity.It is difficult to diagnosis the sudden death from hypersensitivity in the medicolegal expertise.Diagnosis of anaphylaxis is based on the occurrence of characteristic symptoms,especially when they develop upon exposure to a potential trigger.If we diagnose sudden death from hypersensitivity,we must do exclude the death from natural disease,poisoning and other violence,and combine the history of touching anaphylactogens.It is urgent to search a subject and accurate method of diagnosis in clinic medicine and forensic medicine.Therefore,the present study makes the animal mold of anaphylactic shock and sudden death from hypersensitivity with rats and guinea pigs,detects and analyses the content of IgE and P substance in the serum and to make staining in P substance in the lung utilizing enzyme linked immunosorbent assay and radioimmunity and immunohistchemistry respectively;to detect content of tryptase mRNA and of ECPmRNA and make staining in P substance in throat,lung,stomach of rats dead from anaphylactic shock at different time with hybridization in situ.In addition,we detect the serum level of the mast cell tryptase in 115 healthy adults and 15 cases dead from hypersensitivity with fluor euzymelinked immunosorbent assay and survey immunostaining of the mast cell tryptase and P substance in tissue with immunohistochemistry in order to discuss P substance relating to anaphylactic shock and the application value of the level of tryptase,IgE and P substance by measurements in medicolegal expertise.Part 1.The change of IgE,Tryptase and P substance in rats and guinea-pigs of anaphylactic shockObjective To detect the content and change rules of IgE in the serum and IgE immunostaining in the lung of the guinea-pigs in the anaphylactic shock expriments and observe change rules of IgE in the serum at different times after guinea-pig died.To survey tryptase mRNA and ECPmRNA immunostaining in throat,lung,stomach of rats in pre- and post-anaphylaxis.To measure the content of P substance in the serum of the guinea-pigs of anaphylactic shock and observe the immune-state of P substance in throat,lung,stomach of the guinea-pigs of anaphylactic shock.Methods Make the anaphylactic shock model of the guinea-pigs and the rat with respectively the mixed serum of different people and Ovalbumin(OVA).To detect the content of IgE in the serum of the guinea-pigs death from anaphylactic shock utilizing enzyme linked immunosorbent assay,and observe the ch-ange rules of in the serum of the guinea-pigs death from anaphylactic shock in 0h,12h,24h,48h after the guinea-pigs died.To observe expression of IgE in lung of the guinea-pigs death from anaphylactic shock utilizing immunohistochemistry. To survey expression of tryptase mRNA and ECPmRNA in throat,lung,stomach of rats of anaphylactic shock at different times with hybridization in situ.To detect the content of P substance in the serum of the guinea-pigs of anaphylactic shock with radio-immunity.To survey the expression of P substance in throat,lung,stomach of the guinea-pigs of anaphylactic shock with immunohistochemistry.Results(1) The content of IgE in control group＜1 Iu/ml,and the content of IgE have not obvious difference at 12h,24h,48h after the guiea-pigs of control groups died.The content of IgE in the guiea-pigs dead from anaphylactic shock is 13.25±4.32 Iu/ml,15.87±4.66 Iu/ml,12.31±3.13 Iu/ml,14.61±5.46 Iu/ml respectively at 0h,12h,24h,48h after the guiea-pigs died. There is the significant difference between the control group and the experimental group(P＜0.001).The content of IgE is not obvious difference at 12h,24h,48h after the guiea-pigs died from anaphylactic shock(each groups compare P＞0.05).The result of immunohistochemistry dyeing indicates that there are some pale yellow granulas in the lung small vessle wall,circum-vessel and alveolar wall of the experimental groups.It is not change accordingly in the experimental group.To compare with the control group,the average density of IgE in the experimental groups increases significant by analysis with BI-2000 patho-image analysis system(P＜0.001),there is not significant difference in experimental groups(P＞0.05).(2) The content of P substance in the serum of the guinea-pigs of anaphylactic shock is 131.01±18.93pg/μl,the content of P substance in the control groups is 87.70±7.60pg/μl.To compare to the control group,the content of P substance in the serum of the guinea-pigs of anaphylactic shock increases significantly(P＜0.01).Immunohistochemistry expression of P substance in throat,lung,stomach of the guinea-pigs of anaphylactic shock present masculine or hardro-masculine,but immunohistochemistry expression of P substance in throat,lung,stomach of the control group present negative or weakly positive(P＜0.01).(3) The average density of tryptase mRNA and of ECPmRNA in throat,lung,stomach of rats of anaphylactic shock is higher than the control groups.The content of tryptase mRNA in each tissues achieve peak in 30 minutes after allergize,sequentia the content of tryptase mRNA fall-off,still is higher than normal level in 24 hours after allergize.The content of ECPmRNA in throat begin to increase,and maintain a higher level in 30 minutes after allergize,The content of ECPmRNA in lung achieve peak in 6 hours after allergize,The content of ECPmRNA in stomach achieve peak in 24 hours after allergize.Conclusion The content of IgE in the serum have no marked change in 4℃,measuring the serum level of IgE can provide an objective diadynamic criteria to diagnose anaphylactic shock for forensic experts.P substance as the signal materil which can be recognized by nervous system and immune system participate in anaphylactic shock' happening and development.The content of tryptase mRNA in rats of anaphylactic shock can achieve peak earlier than the content of ECPmRNA,tryptase can earlier illustrate the happening of anaphylactic shock.ECPmRNA achieve peak late,it is an important signal of anaphylactic response late onset phasing.Part 2.Study of forensic pathologic diagnosis in human(dead) body sudden death from hypersensitivityObjective To detect the content in the serunm of health adult,eatablish range of normal values.To observe the tryptase'and IgE' content in the serum of dead body who be diagnosed sudden death from hypersensitivity,and to observe the difference of P substance'expression in throat,lung,stomach between health adults and dead bodies died from hypersensitivity in order to approach the diagnosis method of sudden death from hypersensitivity.Methods To detect the serum level of the mast cell tryptase in 115 healthy adults and 15 cases dead from hypersensitivity with fluor euzymelinked immunosorbent assay and to detect the content of IgE in the serum of 7 cases dead from hypersensitivity with enzyme linked immunosorbent assay,and to survey immunostaining of the mast cell tryptase and P substance in throat,lung,stomach of 15 cases dead from hypersensitivity with immunohistochemistry, and to make quantitative analysis with BI-2000 patho- image analysis system. Results(I) The content of Mast cell tryptase in the serum of 115 healthy adults,the nadir is 0.80μg/L,the maximum is 8.86μg/L,the arithmetic mean is 3.06μg/L,the geometric mean is 2.69μg/L,the 95 percentage quantile is 6.00μg/L.There is not obvisous difference between man and woman(P＞0.05).There is not obvisous difference among healthy individuals＜21 years, healthy individuals 21 to 40 years,healthy individuals 41 to 60 years,healthy individuals＞60 years(P＜0.05).(2) The content of tryptase in the serum of 6 cases sudden dead from hypersensitivity is respectively 25.01μg/L,8.91μg/L,19.70μg/L,6.01μg/L,10.15μg/L, 19.52μg/L,the content of tryptase in the serum of 6 cases is higher than the content of tryptase in the serum of healthy individuals.(3) The content of IgE in the serum of 7 cases dead from hypersensitivity there are 6 cases which higher than normal level(＞333U/ml),1 case is within normal(260.82 U/ml).(4) The content of tryptase in throat,lung,stomach of the dead bodies died from hypersensitivity is higher than healthy individual,there is significant difference compare with control group(P＜0.01).PI of P substance in the throat,lung, stomach respectively is 129.839,99.2880,113.6031,PI of the control group respectively is 44.8630,39.5439,68.7888(P＜0.001).Conclusion The normal range of tryptase in the serum of 115 healthy individuals is 0μg/L～6μg/L,there is not obvisous difference between man and woman.There is not obvisous difference among the people whose age are below 60 years,the content in the serum of the people whose ages are above 60 years.The content of tryptase in the serum of the individuals dead from hypersensitivity and the content of tryptase in throat,lung,stomach of the dead bodies died from hypersensitivity is higher than normal level.The expression of P substance in the throat,lung,stomach of the dead bodies died from hypersensitivity enhance.Accordingly, to detect the content of IgE and tryptase in the serum of the dead body can diagnosis sudden death from hypersensitivity,immunohistochemistry dyeing of tryptase in the lung can reflect tryptase'activity and content.In these cases which have no condition to detect IgE and tryptase in the serum but is highly suspected sudden death from hypersensitive,we can observe immunohistochemistry dyeing of tryptase in the lung.