Objective To explore the expression change of intestinal epithelial tight junction (IJ)protein claudin-1 in mice with fulminant hepatic failure (FHF).Methods FHF was induced with a method that combined intraperitoneal injection of lipopolysaccharide (LPS,10 mg/kg) and D-galactosamine (GalN,800 mg/kg).Control saline (2 ml/kg,ip),LPS (10 mg/kg,ip) and GaIN (800 mg/kg,ip) were also detected.The effect of administration of anti-tumor necrosis factor alpha (TNF-α) IgG antibody (anti-TNF-α IgG,100 μg/per) on the level of TNF-α was assessed before administration of D-galactosamine/lipopolysaccharide.At the 2nd h,6th h,9th h,12th h,24th h after injection in FHF group,the 9th h after injection in control groups and 9th h after injection in anti-TNF-α IgG group,the mice were killed for the collection of large intestine specimens.Claudin-1 was analyzed with immunohistochemistry,Western blotting,and real-time quantitative PCR.Results Tight junction protein claudin-1 was localized along the apical region of the lateral plasma membrane representing the region of tight junctions in surface and crypt epithelial cells.Weakly distributed density of claudin-1 in intestinal mucosa was found in mice with FHF from the 9th h after injection.Compared to saline group,Western blotting analysis demonstrated markedly reduced claudin-1 expression in mice with FHF at the 6th h and 9th h after injection (6th h:0.8600±0.0208 vs 1.0,P ＜0.05; 9th h:0.6633 ±0.0328 vs 1.0,P ＜0.01).Furthermore,the expression of claudin-1 mRNA was markedly reduced at the 6th h,9th h,and 12th h after injection in mice with FHF (6th h:0.3067 ±0.1291 vs 1.0,P ＜0.05; 9th h:0.2233 ±0.1155 vs 1.0,P ＜0.01 ; 12th h:0.5275 ±0.1222 vs 1.0,P ＜0.05).Compared to saline group,no significant difference in claudin-1 expression was found with prophylactic treatment with anti-TNF-α-IgG antibody in mice with FHF at the 9th h after injection (protein:0.9533 ±0.0186 vs 1.0,P ＞0.05; mRNA:0.85 ±0.1437 vs 1.0,P ＞0.05).Conclusions The expression of tight junction protein claudin-1 was reduced at both protein and mRNA levels in intestinal epithelial cells that were induced by TNF-α in mice model of FHF.

Objective To investigate the characteristics of early changes in serum IL-17 and IFN-γ levels in elderly patients with acute myocardial infarction (AMI).Methods 70 hospitalized elderly patients with AMI and 35 healthy people were selected.Serum level of interleukin-17 (IL 17) and interferon-γ (IFN-γ) were assayed by enzyme linked immunosorbent assay (ELISA).Results Serum levels of IL17 and IFN-γ showed increasing trends in elderly patients with AMI as compared with that in control group,but there were no significant differences between the two group [(35.73 30.28) pg/ml vs.(28.70±17.12) pg/ml,(15.29±14.64) pg/ml vs.(11.38±10.10) pg/ml,t=0.144 and 0.138,P=0.365 and 0.377].There were correlations between serum IL-17 and IFN γ levels in patients with AMI and controls(r=0.936 and 0.989,both P=0.00).Serum levels of IL-17 or IFN-γhad no significant differences between AMI patients with well and poor prognosis [(35.43± 34.36) ng/L vs.(36.11±30.16) ng/L,(13.90±13.98) ng/L vs.(15.99±14.14) ng/L,U=0.266 and 0.166,P=0.687 and 0.668].Conclusions Serum IL-17 level has an increasing trend in AMI patients within 24h,but has no statistical significant.Serum IL 17 level has a significantly positively correlation with serum IFN γ level in the elderly,but serum levels of IL-17 or IFN γ have no significant correlations with short term prognosis in elderly patients with AMI.

Objective To investigate the correlation between large platelet ratio (P-LCR) and stroke in patients with atrial fibrillation. Methods 152 consecutive patients with atrial fibrillation were selected and divided into two groups according to whether accompanying stoke or not. Clinical features of patients and blood test indicator P-LCR were analyzed by Logistic regression analysis. Results In total, 152 subjects, including 47 patients with stoke and 105 patients without stoke,were included in the study. Univariate analysis showed that the difference in age, diabetes, hypertension, lipid-lowering therapy and P-LCR between two groups was statistically significant (P < 0.05). Logistic regression analysis showed that age, diabetes and P-LCR were independent risk factors of stroke in patients with atrial fibrillation. Conclusions Age, diabetes and P-LCR are independent risk factors of stroke in patients with atrial fibrillation.

Objective:This study aimed to establish an eXtreme Gradient Boosting(XGBoost) model that can predict the recurrence of hepatocellular carcinoma(HCC)patients after laparoscopic hepatectomy (LH) surgery.Methods:A total of 440 patients with primary HCC who received LH treatment for the first time from January 2013 to September 2016 in Affiliated Hospital of Chengde Medical University were selected as the research objects. The diagnosis method was pathological diagnosis. Research objects were divided into training group ( n=88) and verification group ( n=352) at a ratio of 2∶8 by random number table method. The Kaplan-Meier method was used to draw the recurrence-free survival curve, and the Log-rank test was used to compare the survival of the two groups; the training group was used to establish the COX regression model and the XGBoost model to screen independent predictors of recurrence after LH; receiver operating characteristic(ROC) curve was used to analyze the predictive abilities of the two models, and conducted internal verification in the verification group; Hosmer and Lemeshow Test was used to evaluate the calibration of the two models, and P>0.05 was used as a good fit between the model and the actual situation. Results:Both the COX regression model and the XGBoost model screened out tumor thrombus, low degree of differentiation, tumor microvascular infiltration (MVI), number of tumors, large tumors, and positive hepatitis B surface antigen were independent predictors of tumor recurrence( HR=2.477, 0.769, 1.786, 1.905, 1.544, 1.805; 95% CI: 1.465-4.251, 0.619-0.819, 1.263-2.546, 1.354-2.704, 1.272-1.816, 1.055-2.555). The XGboost model scores were 32 points, 29 points, 24 points, 18 points, 16 points, 11 points, respectively. In the training group, the area under the curve (AUC) of the COX regression model and XGBoost model for predicting recurrence were 0.746 (0.730-0.762) and 0.802 (0.785-0.818), respectively. The XGBoost model had strong predictive ability and was confirmed in the validation cohort. Conclusions:This study had established and verified the XGBoost model that can predict the recurrence of HCC patients after receiving LH for the first time. It can be used in clinics to assist doctors in formulating personalized postoperative monitoring programs for patients. Early detection, early diagnosis and early treatment of tumors and strengthening of postoperative follow-up are important measures to improve the prognosis of patients.

Objective To express human-mouse chimeric antibody against anthrax protective anti-gen and to analyze its biological activities. Methods A new mammalian bipromoter expression vector was constructed with dihydrofolate reduetase(DHFR) gene as the selection and complication marker. First, the light and heavy chain variable region gene of the monoclonal antibody 5E1 were cloned by RT-PCR, at the same time the human IgG1 heavy chain constant region gene and kappa type constant region gene were cloned. Next, the human-mouse chimeric antibody genes were synthesized by fusion PCR. Then, the hu-man-mouse chimeric antibody gene were inserted into MCS of pSecTag and B1 to construct pSecTag-5E1L and B1-5E1H, respectively. Finally, heavy chain expression cassette excised from the B1-5E1H with Bgl Ⅱ/BamH Ⅰ was further cloned into the Bgl Ⅱ site of the pSecTag-5E1L to construct pSecTag-5E1. Plasmid pSecTag-5E1 was transfected into CHO(dhfr) engineering cells and high production cell clones that were screened by enhancing MTX concentration. After collecting medium and purifying chimeric antibody with af-finity chromatogram, purified chimeric antibody was analyzed by SDS-PAGE, Western blot. Results A sta-ble and high production cell line was acquired at MTX concentration 5×10~(-8) mol/L. Conclusion The hu-man-mouse chimeric antibodies were successfully expressed in CHO cells.

Objective: To analyze whether heat-shock protein 90 (HSP90) be involved in a permanently abnormal activated JAK/STAT signaling in ATL cells in vitro. Methods: The effect of 17-AAG on proliferation of ATL cell lines HUT-102 was assessed using CCK8 at different time points. Cell apoptosis was measured by flow cytometry. The specific proteins HSP90, STAT5, p-STAT5 and JAK3 were detected by Western blotting. Results: Overexpression of HSP90 in HUT-102 cell lines was disclosed (P<0.05) , and constitutive activation of JAK3/STAT5 signaling was observed in HTLV-1-infected T-cell lines but not in normal PBMCs; Treatment of ATL cell lines with 17-AAG led to reduced cell proliferation, but there was no significant change in terms of cell proliferation when the concentration of 17-AAG between 2 000-8 000 nmol/L (P>0.05) . 17-AAG induced cell apoptosis in different time-points and concentrations. 17-AAG don’t affect the expression of JAK3 gene. Conclusion This study indicated that JAK3 as HSP90 client protein was aberrantly activated in HTLV-1-infected T-cell lines, leading to constitutive activation of p-STAT5 in JAK/STAT signal pathway, which demonstrated that HSP90-inhibitors 17-AAG inhibited the growth of HTLV-1-infected T-cell lines by reducing cell proliferation and inducing cell apoptosis.

Severe frostbite (grade Ⅲ to Ⅳ) is a common disease accompanied with high disability rate in cold regions, especially for military training and disaster events in cold regions. The treatment of severe frostbite mainly includes rapid rewarming in the early stage and amputation in the later stage; while the damage of vascular endothelial cells, microvascular thrombosis, and decreased tissue perfusion secondary to severe frostbite are important factors affecting prognosis. Transcatheter arterial thrombolysis is a new technique for the treatment of severe frostbite. It has the advantages of minimally invasive, high safety, and significantly reduced amputation rate. We reviewed the advances in the research of transcatheter arterial thrombolysis for the treatment of severe frostbite.

Objective To avoid misdiagnosis of adult abdominal epilepsy. Methods Analyzed the clinical data of a misdiagnosed adult patient with 16 years of abdominal epilepsy, also reviewed the related literatures. Results Both adult abdominal epilepsy and the misdiagnosis of it for 16 years were very rare. Differential diagnosis of it should be used to distinguish with a variety of diseases. Conclusion It is crucial to avoid misdiagnosis of the disease that exam-ining patient with electroencephalogram (EEG) at the onset of abdominal pain and thinking carefully with detailed and comprehensive analysis. Once the diagnosis is definite, antiepileptic therapy should be applied.

OBJECTIVEA RP-HPLC method was developed for simultaneous determination of bigelovin, ergolide and tomentosin in Inula hupehensis.

METHODAn Agilent C18 column (4.6 mm x 250 mm, 5 microm) was used for separation at 40 degrees C. The mobile phase was acetonitrile-water, and the flow rate was 1.2 mL x min(-1). The detection wavelength was set at 210 nm.

RESULTThe method has good linearity in the ranges of 0.01792-0.1792 g x L(-1) (r =0.9999) for bigelovin, 0.0424-0.4240 g x L(-1) (r =0.9996) for ergolide, and 0.044 8-0.4480 g x L(-1) (r = 0.9996) for tomentosin. The average recoveries of bigelovin, ergolide, and tomentosin were 98.5%, 98.2%, 98.4%, with the RSD of 1. 3%, 1.3%, 1.7%, respectively. The results demonstrated that there was a significant difference in the contents of three sequterpene lactones among the tested Inulae Flos.

CONCLUSIONThe results indicated that the present RP-HPLC method is simple, quick and accurate, and can be used for the quality control of I. hupehensis, especially for the authentication of Inulae Flos.

Objective To evaluate the long-term efficacy and safety of super-selective hepatic artery embolization (TAE) and bleomycin hydrochloride in treatment of liver hemangioma.Methods A retrospective analysis was conducted on 132 patients treated for liver hemangioma from January 2011 to December 2014.The patients underwent super-selective TAE.There were 26 men and 106 women.The age ranged from 29.0 to 65.0 (46.0±8.0) years.An emulsion of iodized oil and bleomycin and polyvinyl alcohol particle (PVA) was used for the TAE.Routine follow-up visits on post-treatment Days 3 and 7 were carried out to detect complications and changes in liver function.The diameters of the lesions pre-TAE and at 6,12 and every 12 months thereafter were monitored.Results TAE was technically successful in all these patients.The range of follow-up was 3～6 years.The successful rates of treatment at 6 month,12 months,24 months,and 36 months,were 100％,100％,96.9％ and 90.6％,respectively.Conclusion TAE with an emulsion of iodized oil and bleomycin and PVA was a safe and efficacious treatment for liver hemangioma on long-term follow-up.