The Expression Of Toll-like Receptor 4 In The Guinea Pig Died Of Anaphylactic Shock And Forensic Significance

Objective: Establishing guinea pigs anaphylactic Shock model,and making specimens of tissue and blood serum.To detect the expression of guinea pig Ig E and TLR4.Trying to lay the foundation for the experimental model and to find new objective diadynamic criteria to diagnose anaphylactic shock for forensic.Methods: 35 healthy guinea pigs of either sex,weighing 220-300 g,classified into the experimental group of 26 and the control group of 9 with no purpose.The experimental group were immunized intracutaneously with human mixed serum and 3 days were immunized again,then induced by endocardiac injection after 2 weeks.The control group injected with saline and Put to excution by fracture cervical spinal cord.Clinical manifestations observed after three-injection of the guinea pig.Perform autopsies on two groups of guinea pigs.Blood was drawn from heart and the serum was purified.Trachea,lungs,stomach and spleen of guinea pigs were taken out,then fixed in 4% polyoxymethylene and embedded in paraffin or put in RNAstore.ECLIA method to detect the expression of guinea pig serum Ig E.RT-PCR and IHCI methods to detect the expression of guinea pig tissue TLR4.SPSS 20.0 software was used to analyze experimental data.Results:(1)Experimental groups all guinea pigs appeared the typica l clinical symptoms of anaphylactic shock after the injection human mixed serum.18 guinea pigs decease within 2-30 minutes;Control groups showed none of these complication.(2)Experimental group guinea pigs were observed mucosal edema,blood congestion in multiple visceral organs with HE staining in anaphylactic shock;And eosinophilia in trachea,lungs,stomach and spleen tissues were observed.Control group showed none of these expressions.(3)The Ig E Ievels of serum in experimental group is higher than that in control group,P<0.05;(4)The TLR4 levels of trchea,lungs,stomach and spleen tissues within RT-PCR method in experimental group is higher than that in control group,P<0.05.(5)The TLR4 levels of trchea,lungs,stomach and spleen tissues within IHC method in experimental group is higher than that in control group,P<0.05.Conclusion:(1)Successfully established a guinea pig in anaphylactic shock animal models.(2)The reference value of the serum Ig E in the sudden death of anaphylactic shock in the identification of the forensic medicine was verified.(3)RT-PCR and IHC measurement of TLR4 in trachea lungs,stomach and spleen tissues can assist in diagnosis of anaphylactic shock.