The Expression Of Relative Mediators In The Tissue Of Guinea Pigs Died Of Anaphylactic Shock And Medicolegal Significance

IntroductionAnaphylactic shock belongs to the IgE-mediated immediate allergy which is urgent and dangerous. Anaphylactic shock can show dermal, respiratory, digestive systems and cardiovascular system symptoms, and severe cases can died of anaphylactic shock. Some of the cases can cause civil procedure and medical tangle.The bodies died of anaphylactic shock are often without specific pathological changes and lacking in objective diagnose indicators. According to the mechanism of anaphylactic shock, the various mediators which degranulate from mast cell play an important role in anaphylactic shock. The tryptase and chymase which belong to neutral serine protease are stable, and its molecular weight is bigger than other mediators. Thus, they are both the signs of mast cell degranulated.IL-5 is one of the chemokines of eosinophil(EOS), which is the main effect factor in allergic type I . Its study is focus on the asthma. At present, there is no research in anaphylactic shock.This study is on the basis of animal models of guinea pigs who died of anaphylactic shock, using immunohistochemistry(IHC) technology, observe the expression of these mediators. Then analysis is made to evalue medicolegal significance by using the methods of pathology and statistics, depending on Malpractice Bill. Finally, the aim of this study is providing the objective evidences to the diagnosis of anaphylactic shock. Materials and Methods1. Establishment of animal model and packetA total of 24 healthy guinea pigs are divided into 0 hour group, 48 hours group in 4℃, and 7 days group in -20℃at random, 8 in each group. Each group is also divided into experimental group and control group, 4 in each group. The guinea pigs in experimental group evoke anaphylactic shock, then died after injecting human mixed serum. The guinea pigs in control group executed by ether poisoning after injecting NS. Blood was drawn from heart and the serum was purified. Larynx, trachea, lungs, and stomach of guinea pigs were taken out, then fixed in 4% polyoxymethylene and embedded in paraffin. Immunohitochemistry of tryptase and chymase was performed in 5 cases died of anaphylactic shock and 5 cases died of the other reasons.2. Methods(1) HE staining.(2) Stain EOS by using the method of Carbol Chromotrope 2R staining.(3) Detect of the IgE levels in serum by using chemiluminescent.(4) Tryptase, chymase and IL-5 immunohistochemical staining.(5) Calculate and analyze positive expression by Motic Image Advanced 3.2 system.(6) Analyze the differences between groups using SPSS.Result(1) The guinea pigs' larynx and trachea were of edema, pulmonary alveolus were of expansion, and alveolar wall became thin with HE staining in anaphylactic shock.(2) There was a number of eosinophils in tissue by Carbol Chromotrope 2R staining in experimental group.(3) The IgE levels of serum in experimental group is higher than that in controlgroup, P<0.01.(4) There are scattered tryptase and chymase positive mast cells by IHC method in control groups, and brown staining product in cytoplasm. The positive cells number of tryptase and chymase increased and the expression enhanced, and staining product partly diffused around mast cells and in tissue space in experimental groups. There were scattered positive cells of IL-5 in control group, staining product in cytoplasm. In experimental group, positive cells staining product mostly diffused around cells and in tissue space. There were differences between experimental and control group by SPSS.(5) The tryptase and chymase positive granules of non-anaphylactic shock in actual cases were seen in cytoplasm of mast cells, and mast cells have complete form. The most staining product diffused around cells and in tissue space, and mast cells have incomplete form in anaphylactic shock cases.DiscussionAnaphylactic shock belongs to the IgE-mediated immediate allergy which is urgent and fatal to the human, therefore, it is met in forensic identification ususlly.Because of the stimulater by antigen, body can produce much IgE, when the same antigen come in the body again, the FcεRI on the face of mast cells which have been sensitized can activate by neighboring IgE. Fanally, mast cells degranulate, and show up clinical symptoms.The people who died of anaphylactic shock, the IgE levels of serum usually higher than the normal, the number of eosinophils in tissue increased. But, the IgE levels usually effected by external factors, while the eosinophils increased is not the unique apparence, so both of them are lack of objectivity in medicolegal expertise.There is a pathological phenomenon, that is the mast cells degranulate in anaphylactic shock. How to prove if mast cells degranulate can provide a strong evidence to the diagnosis of anaphylactic shock. This study just to find the evidence of mast cells degranulate. Tryptase and chymase are both neutral mediators in mast cell, which are very stable. They can spread to tissue space accompanied by mast cell degranulate. IL-5 is the chemokines of eosinophils. In some diseases which register as eosinophils increased, the expression of IL-5 will be stronger. This study by comparing the experimental groups with control groups'expression of these mediators, prove that they were stronger in the guinea pigs who died of anaphylactic shock. Further more, tryptase and chymase can reflect if mast cells degranulate. Therefore, it is can say that use the medthod of IHC to observe the expression of tryptase, chymase and IL-5 will provide new evidence to the diagnosis of anaphylactic shock.When have this method application in the actual cases, discover that the expression of tryptase and chymase stronger in anaphylactic shock group than that in non-anaphylactic shock group, The tryptase and chymase positive granules of non-anaphylactic shock in actual cases were seen in cytoplasm of mast cells, and mast cells have complete form. The most staining product diffused around cells and in tissue space, and mast cells have incomplete form in anaphylactic shock cases. So, this method of IHC can be applied to the diagnosis of actual cases to distinguish the anaphylactic shock and others.Conclusion1. The guinea pigs' larynx and trachea were of edema, pulmonary alveolus were of expansion, and alveolar wall became thin with HE staining in anaphylactic shock.2. There was a number of eosinophils in tissue by Carbol Chromotrope 2R staining in experimental group.3. The IgE levels of serum in experimental group is higher than that in control group.4. The expression of tryptase, chymase and IL-5 of guinea pigs in anaphylactic shock enhanced, which can provide diagnostic evidences of morphology to anaphylactic shock.5. According to the condition set by Malpractice Bill, there is no evidently effection about using IHC to observe these three mediators.6. The IHC staining of tryptase and chymase is expected to be applied in anaphylactic shock cases, providing diagnostic evidences.