Objective To express a recombinant protein TFPR1 ( the functional region of the snake venom proteins from Trimeresurus flavoviridis) in Pichia pastoris expression system. Methods The target gene was codon-optimized and synthesized according to the sequence of the conserved structural do-main of triflin and then cloned into the Pichia pastoris expression vector pPICZαA to construct the recombi-nant expression plasmid pPICZαA-TFPR1. The recombinant plasmid pPICZαA-TFPR1 was electroporated into the yeast strain X33. The transformed strains carrying expression plasmid were screened out with Zeocin and then induced by methanol to express the recombinant protein TFPR1. ELISA was performed for the screening of positive clones. SDS-PAGE and Western blot were used for further identification of the ex-pressed products. Results The recombinant plasmid pPICZαA-TFPR1 was successfully constructed. The recombinant protein TFPR1 was expressed in a secreted form at a molecular weight of 16×103. Conclusion The recombinant protein TFPR1 was successfully expressed in Pichia pastoris expression system, which laid a foundation for further researches on its biological function and application as an adjuvant.

Objective To obtain Chinese hamster ovary (CHO) cell lines that stably express a targeting complement inhibitor CR2-CD59.Methods The recombinant plasmid PEE14.1-CR2-CD59 was constru-cted by cloning the DNA fragment CR2-CD59 into plasmid PEE14.1,and the obtained plasmid was transfected into CHO cells by FuGENE 6.The clones with stable high expression of target fragment were selected by methionine sulfoximine (MSX),the expression of CR2-CD59 was analyzed by ELISA,SDS-PAGE and Western blotting analysis.Results Several stable expression clones were obtained,and CR2-CD59 was highly expressed in the secret form in CHO cells.SDS-PAGE analysis showed that the molecular weight of the recombined protein CR2-CD59 was consistent with the predicted one.ELISA and Western blotting results revealed that the CR2-CD59 could react with both anti-human CR2 and anti-human CD59 polyclonal antibodies.Compared with serum-containing medium,the protein was highly expressed in serum-free medium (P

BACKGROUND/AIMS: High-fat diets contribute to pancreatic fibrogenesis, but the pathogenesis remains unclear. This study investigated the role of nuclear factor kappa B (NF-kappaB) in high-fat diet-induced pancreatic fibrosis in rats. METHODS: Male Wistar rats were fed a high-fat diet or standard normal chow for 20 weeks. Pancreatic fibrosis was determined by Sirius red staining. Immunohistochemical staining, reverse transcription-polymerase chain reaction and Western blotting were used to identify NF-kappaB-associated genes or protein expressions. RESULTS: Inflammation, fat deposition, pancreatic stellate cell activation and fibrosis were observed in the pancreases of the high-fat diet group. NF-kappaB subunit p65 (NF-kappaB/p65) expression was localized to the nucleus, and intercellular adhesion molecule 1 (ICAM-1) was over-expressed. Pancreatic gene expression levels of NF-kappaB/p65, ICAM-1 and tumor necrosis factor alpha were all elevated significantly in rats fed a high-fat diet compared with control rats. Western blotting also revealed significantly increased levels of ICAM-1 and nuclear NF-kappaB/p65 in rats fed high-fat diets comparison with control rats. CONCLUSIONS: NF-kappaB is involved in high-fat diet-related pancreatic fibrosis.
Animals ; Blotting, Western ; Diet, High-Fat ; Fibrosis ; Gene Expression ; Humans ; Inflammation ; Intercellular Adhesion Molecule-1 ; Male ; NF-kappa B ; Pancreas ; Pancreatic Stellate Cells ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha

OBJECTIVETo evaluate the effect of autograft bone, allograft bone, calcium sulfate bone cement, and calcium phosphate bone cement on the repair of tibial plateau defect in rabbits.

METHODSWe used autograft bone, allograft bone, calcium sulfate bone cement, and calcium phosphate bone cement to repair tibial plateau defect in rabbits. Gross and histologic observations, X-ray examination, and biomechanical test were conducted at 1, 2, 4, 8 weeks after operation.

RESULTSX-ray examination found that the bone density was evidently reduced in calcium sulfate group at 8 weeks after operation; there were no marked changes in other groups. The maximal load measurements showed that autograft and allograft groups were greater than calcium sulfate and calcium phosphate groups at 1 and 2 weeks after operation. However at 4 and 8 weeks after operation, no significant difference was found among the four groups. In autograft and allograft groups, there was no significant difference in biomechanical intensity at 2, 4, and 8 weeks, but it was significantly higher than that at 1 week. In calcium sulfate and calcium phosphate groups, the outcome was ranked in descending order as 1 week less than 2 week less than 4 week equal to 8 week. Histologic examination found a great amount of new bones at 8 week in both autograft and allograft groups. In calcium sulfate group, calcium sulfate was almost absorbed and there were numerous bone trabeculations. There was a large amount of unabsorbed calcium phosphate in calcium phosphate group.

CONCLUSIONAt 1-2 weeks postoperatively, the biomechanical intensity is higher in autograft and allograft groups than calcium sulfate and calcium phosphate groups, but after 4-8 weeks, there is no significant difference among groups. At 1-2 weeks, the biomechanical intensity in all groups is increased, but at 4-8 weeks, there is no significant increase. The rates of absorption and bone formation are quicker in calcium sulfate group than calcium phosphate group.

Animals ; Biomechanical Phenomena ; Bone Cements ; Bone Transplantation ; Knee Joint ; Tibia ; Transplantation, Autologous

OBJECTIVETo explore the current status of HIV infection and HIV-related knowledge, attitude and practice (KAP) among men who have sex with men (MSM) in 14 cities in China.

METHODSA cluster sampling method was administrated and 14 large or medium cities were selected. The respondent-driven sampling (RDS) was also used to conduct a questionnaire survey in 6101 MSM. Blood specimens were collected for HIV and syphilis testing.

RESULTSA total of 6101 MSM were selected and investigated, with an HIV infection rate of 6.3% (384/6101) and a syphilis infection rate of 14.0% (854/6101). The proportion of MSM knowing correct HIV/AIDS knowledge was 59.7% (3644/6101); 6.2% (378/6101) and 11.2% (683/6101) of the respondents ever had buying sex or selling sex in the last six months respectively; the proportion of consistent condom-using each time while with male partners in the last month was 39.2% (2392/6101). The proportion of condom-using while with male partners was 65.2% (3978/6101) in the last time among MSM.

CONCLUSIONA high HIV infection rate was observed among MSM with limited HIV/AIDS awareness and high risk behaviors. It is necessary to initiate a rapid response to prevent HIV spreading in MSM.

Acquired Immunodeficiency Syndrome ; epidemiology ; China ; epidemiology ; Cluster Analysis ; Health Knowledge, Attitudes, Practice ; Homosexuality, Male ; psychology ; Humans ; Incidence ; Male ; Risk-Taking ; Surveys and Questionnaires ; Unsafe Sex

Objective To analyze and compare the pathological changes of lung tissue in mice infected with the novel H7N9 influenza virus and 2009 pandemic H1N1 influenza virus, respectively, and to preliminarily study the mecha-nisms of acute lung injury induced by those virus infection .Methods SPF 6-week old BALB/c mice ( body weight 18-20 g, male∶female=1∶1) (n=3 in each subgroup) were intranasally infected with H7N9 virus and H1N1 virus, respec-tively.The behavior and survival time of mice after virus infection were observed and the survival rates were analyzed .The heart, liver, spleen, lung, kidney, intestines, and brain were collected at indicated time points for histopathological exami-nation using H&E staining .The distribution of virus antigen was detected by immunohistochemistry .The neutrophil infiltra-tion was also observed .The correlation of lung injury with virus replication and host immune responses was analyzed .Re-sults The lung and spleen injury of mice infected with H 7N9 virus was slighter and their survival rate (100%) was high-er than those of mice infected with H1N1 virus.The damages of the lung and spleen in H1N1virus-infected mice were more severe than that in H7N9 virus-infected mice, and all the 10 mice in this group died within 9 days after virus inoculation . The distributions of both the virus antigens were mainly in the bronchial epithelial cells , a few stromal cells and alveolar ep-ithelial cells .The levels of virus replication in the two groups were not significantly different .There were more intense neu-trophil infiltration in the lung and inflammatory response in the H 1N1 virus-infected mice than those in the H7N9 virus-in-fected mice .Conclusions There are some differences of the pathological characteristics and extent of lung injury in the mice infected with H7N9 virus and H1N1 virus, respectively.The virus replication is a precipitating factor but not the deci-sive factor of the lung injury , and there is a close relationship between the host immune responses and acute lung injury .

Objective: To evaluate the effect of autograft bone,allograft bone,calcium sulfate bone cement,and calcium phosphate bone cement on the repair of tibial plateau defect in rabbits.Methods: We used autograft bone,allograft bone,calcium sulfate bone cement,and calcium phosphate bone cement to repair tibial plateau defect in rabbits.Gross and histologic observations,X-ray examination,and biomechanical test were conducted at 1,2,4,8 weeks after operation.Results: X-ray examination found that the bone density was evidently reduced in calcium sulfate group at 8 weeks after operation; there were no marked changes in other groups.The maximal load measurements showed that autograft and allograft groups were greater than calcium sulfate and calcium phosphate groups at 1 and 2 weeks after operation.However at 4 and 8 weeks after operation,no significant difference was found among the four groups.In autograft and allograft groups,there was no significant difference in biomechanical intensity at 2,4,and 8 weeks,but it was significantly higher than that at 1 week.In calcium sulfate and calcium phosphate groups,the outcome was ranked in descending order as 1 week＜ 2 week＜ 4 week =8week.Histologic examination found a great amount of new bones at 8 week in both autograft and allograft groups.In calcium sulfate group,calcium sulfate was almost absorbed and there were numerous bone trabeculations.There was a large amount of unabsorbed calcium phosphate in calcium phosphate group.Conclusion: At 1-2 weeks postoperatively,the biomechanical intensity is higher in autograft and allograft groups than calcium sulfate and calcium phosphate groups,but after 4-8 weeks,there is no significant difference among groups.At 1-2 weeks,the biomechanical intensity in all groups is increased,but at 4-8 weeks,there is no significant increase.The rates of absorption and bone formation are quicker in calcium sulfate group than calcium phosphate group.

Objective To investigate the clinical application of modified electroconvulsive therapy (MECT),and provide the references for more standardized and rational use of MECT.Methods A retrospective study was performed by stratified random sampling method.The clinical data of 1 330 psychiatric inpatients from January 2006 to December 2015 in Henan Psychiatric Hospital were analyzed retrospectively,and the clinical application of MECT was investigated,including the proportion of psychiatric inpatients receiving MECT,the situation of combined use of MECT and antipsychotic drugs,and the routine examination results before and after MECT.Results The proportion of receiving MECT in schizophrenic patients was the highest,it is 11.36％ (81/713),the second was depressive episode (5.61％,12/214),followed by dissociative conversion disorder (5.56％,2/36),bipolar disorder (5.34％,7/131),organic mental disorder (3.23 ％,1/31) and other mental disorders (2.93％,6/205).There were significant differences in the distribution of various mental disorders between the non electroshock group and the electroshock group (x2 =21.728,P ＜ 0.05).The proportion of the patients without electric shock and the patients with single drug,two drugs,triple therapy or quadruple therapy was 0.92％,17.43 ％,56.88％,21.10％ and 3.67％,respectively.The proportion of the patients with two or more antipsychotic drugs was 81.65％ in the electroshock group.The antipsychotics were olanzapine (28.44％),clozapine (27.52％),quetiapine (26.61％) and risperidone (24.77 ％) according to the drugs used frequently in the electroshock group.The results of routine blood test were compared among the time points of before treatment,the 1st,2nd,3 rd week of MECT and 1 week after MECT in the shock group,which showed that there was significant difference in white blood cell count (F =2.570,P ＜ 0.05);but there was no significant difference in the levels of platelet,hemoglobin and red blood cell (P ＞ 0.05);the level of white blood cell returned to normal within two weeks in 93.33％ patients with increased white blood count.The myocardial enzymes and liver function tests were not perfect during the treatment of MECT and after treatment.There was no long-term systematic evaluation of cognitive function and quality of life in patients with electroconvulsive shock.Conclusions The probability of using MECT in the patients with schizophrenia is more than that in the patients with depression.MECT is used in combination with two or more antipsychotic drugs.MECT may cause a short increase in white blood cells,but the white blood cell level in most patients can recover to normal levels after symptomatic treatment.The clinical data for treatment and prognosis in patients with MECT are inadequate,so it is necessary to improve the medical record.

Objective This study assessed the main factors associated with HIV infection among men having sex with men(MSM),in order to provide evidence for future intervention. Methods A case-control study was conducted among HIV cases identified at the MSM survey in Nanjing, Suzhou and Yangzhou cities from May to July in 2008. Each HIV case was matched by 4 HIV negative MSM who participated consecutively right after the HIV case was identified at the same survey, same time and in the same city. A national core MSM survey questionnaire was used with addition of variables of local interest. Fitting conditional logistic regression model was applied for univariate and multivariate analysis. Results During the survey, 62 HIV positive cases were identified and 248 controls were matched. Data from univariate analysis showed that, inconsistent condom use, syphilis infection, self reported sexually transmitted infection (STI) symptoms and/or signs, unprotected anal sex (UAI) were risk factors, with OR values of 5.56(95%CI: 1.82-16.96), 2.19(95%CI: 1.18-4.05), 2.50(95%(CI: 1.13-5.50) and 3.13 (95%CI: 1.64-5.98) respectively. Online cruising and receptive anal sex were protective factors with OR values as 0.47(95%CI: 0.22-0.998) and 0.67(95% CI: 0.23-1.97). Through multivariate analysis, it was found that syphilis, age of having experienced first insertive sexual intercourse and UAI were risk factors to HIV transmission, with OR values of 2.28 (95%CI: 1.05-4.94), 4.16 (95%CI: 1.63-10.66) and 1.11 (95% CI: 1.04-1.18) respectively. Compared with non-anal sex, bisexual (OR=0.41,95%CI: 0.18-0.97) behavior seemed a protected factor. Conclusion Syphilis infection, UAI and self reported STI symptoms and/or signs were risk factors related to HIV transmission while bisexual seemed a protective factor.

Objective To explore the effect of clinical nursing pathway on the patients with enterostomy.Methods Totals of 36 patients with enterostomy were randomly divided into experimental group and control group,and the experimental group received the nursing of the clinical nursing pathway while the control group received the routine nursing.The complications of enterpstomy,the patient satisfaction,the average hospitalization days and the expenses were observed and compared between two groups.Results There was significantly difference between experimental group and control group in the complication rate (0％ vs 27.77％ ),the length of the mean hospitalization ( 16.12 ± 5.46 d vs 21.18 ± 4.88 d),and the expenses (20325.68 ±5676) vs (22348.18 ±4 479.90),(x2 =4.587 8,t =-2.88,-3.372,respectively;P ＜0.05 ).Patients' satisfaction of experimental group (94.44％) was significantly higher than that of control group (55.55％),( x2 =7.251 7,P ＜ 0.05 ).Conclusions The application of clinical nursing pathway can reduce the complications of enterostomy and patients' sufferings,shorten the mean hospitalization,reduce the expenses,improve the nursing quality and patients' satisfaction.