Objective To study the technical method of immunohistochemical staining in pathological tissue.Methods 80 cases of hospitalized patients in our hospital from May 2015 to May 2016 were selected, they were randomly divided into immunohistochemical staining group(immunohistochemistry group, n=40)and biopsy group(n=40)two groups, then the AFP(alpha-Fetoprotein), Glypican-3 positive situation of the the two groups and diagnostic Resultsof patients with different types of disease in the immunohistochemistry group were statistically analyzed.ResultsThe positive rate of AFP(a-Fetoprotein), Glypican-3 of the immunohistochemistry group 92.5%(37/40)was significantly higher than the biopsy group 50.0%(20/40)(P<0.05), Among the 37 cases of positive patients, metastatic tumors 19 cases, hepatocellular carcinoma 9 cases, intrahepatic bile duct carcinoma 7 cases, hemangioma 3 cases, the positive rates were 100.0%, 75.0%, 100.0%, 100.0%, 26 cases died, the mortality rate was 70.3%, of which metastatic tumors 13 cases, hepatocellular carcinoma 6 cases, intrahepatic bile duct carcinoma 5 cases, hemangioma 2 cases, the mortality rates were 72.2%, 66.7%, 71.4%, 66.7%.Conclusion Immunohistochemical staining technic in pathological tissue can promote positive rate, and standard operation procedures can reduce errors, which is worthy of clinical attention.

Objective To explore the clinical effect of microgranular fat obtained by mechanical injection of autologous fat tissues in the treatment of lacrimal trough deformity.Methods Ten patients in this group were women,and the average age was 28 years.With tumescent technique,the thigh fat granules were aspirated;after the separation and purification using cell sorting screen,fat particles were obtained with uniform size and without fibrous tissue.Through mechanical pushing of the fat particles,the fat granules became chylomicron-like.The fat particles without fibrous tissue were injected into the superficial layer and deep layer of the orbicularis oculi muscle with depressed deformity.3-12 months after operation,the clinical efficacy was evaluated by doctors,patients and the third party.Results In 10 cases of lacrimal groove deformity,9 cases were transplanted once,and 1 case did twice.After 3 to 12 months follow-up the appearance of tear trough deformity was significantly improved,the texture of the ditch of eyelid and cheek skin was soft,and appeared young plump appearance.There were no complications,such as infection,hematoma,fat liquefaction,pigmentation and induration,and satisfactory results were obtained.Conclusions Ultrafine particles of fat are obtained using improved purification method in the treatment of lacrimal groove deformity,and the efficacy is satisfactory.

Objective To investigate the factors affecting the susceptibility of tigecycline and assess the testing methods.Methods The 116 isolates of Acinetobacter baumanaii and Klebsiella pneumoniae were collected in 13 hospitals from January to December,2010,to evaluate the effects on the tigecycline susceptibility of the overnight medium,medium brand and lot number,respectively.The 56 isolates of Acinetobacter baumannii and the 47 isolates of Klebsiella pneumoniae were selected randomly according to the MIC distribution proportion in 2010 and 2012.The broth microdilution was taken as the reference method to evaluate the effects of the agar dilution,disk diffusion,MIC Test Strip (MTS) and Vitek2 (GN16) on the susceptibility of tigecycline.Results The essential agreement (EA) of Acinetobacter baumannii and Klebsiella pneumoniae is 89.7％ (52/58)and 87.9％ (51/58) using overnight medium and fresh medium respectively.Both EA and categorical agreement (CA) of the different brands (BBL and Oxoid) and lot numbers are 100％ using agar dilution.According to the FDA break point criteria,the CA/EA is 77.7％ (80/103)/99.0％ (102/103),87.4％ (90/103)/98.1％ (101/103),64.1％ (66/103)/76.7％ (79/103) using agar dilution,MTS,Vitek (GN16) with respect to broth microdilution.The CA is 79.6％ (82/ 103,S≥14 mm,R≤10 mm),69.9％ (72/103,S≥ 16 mm,R≤ 12 mm),34.0％ (35/103,S≥19 mm,R≤14 mm)using disk diffusion method compared with broth microdilution (FDA break point criteria).Conclusions The susceptibility of tigecycline must be tested using fresh medium.The medium brands and lot numbers used in this test have no effects on the tigecycline susceptibility to Acinetobacter baumannii and Klebsiella pneumoniae.There exist the better correlations on MIC using agar dilution and MTS than the disk diffusion and Vitek(GN16) compared with broth microdilution.It is expected that the consistency can be improved by adjusting the break point of disk diffusion.

AIM: To establish an animal model of experimental autoimmune myocarditis (EAM) in BALB/c mice and to investigate the expression and significance of Toll-like receptor 3 in mouse EAM. METHODS: BALB/c mice were immunized with cardiac myosin extracted from porcine ventricular myocardium covered by complete freund's adjuvant (CFA) on 0 d and 7 d, then divided into immunized with CFA only. Serum and myocardium samples were collected at 14 d and 21 d after the first immunization. HE staining was used to identify the areas of inflammation. The myosin IgG antibody was examined by indirect ELISA assay. The changes of TLR3 protein and mRNA expression in myocardial tissue were measured by immunohistochemistry and real time-PCR. RESULTS: Compared to control group, immunohistochemistry results showed that there was positive expression of TLR3 in the myocardium of mice with EAM and the mRNA of TLR3 were more than 20 times (P<0.05). The expression of interferon beta mRNA in EAM group was more than 14 times as many as basal expression, that of tumor necrosis factor alpha was more than 18 times (P<0.05). CONCLUSION: The expression of Toll-like receptor 3 in myocardium is up-regulated in experimental autoimmune myocarditis. The inflammatory response to cardiac myosin may associate with the TLR3 signal transduction pathway.

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