Objective To investigate the factors related to negative conversion of HBsAg in patients with acute hepatitis B (AHB).Meth-ods A total of 106 AHB patients who were admitted to the Department of Infectious Diseases,Zhoukou Central Hospital from February 2007 to February 2013 were recruited.Liver function,five serological markers of HBV infection (HBsAg,HBeAg,anti-HBs,anti-HBe,and anti-HBc),and HBV-DNA were measured every three months.All patients were followed up for 12 months.The major HBV genotypes (A through D)were determined by type-specific primer nested PCR.Categorical data were expressed as rates and compared with χ2 test;continuous data were expressed as mean ±SD and analyzed by t-test.Results When just admitted to our hospital,compared with the pa-tients with negative conversion of HBsAg,AHB patients with HBsAg persisting for more than 6 or 12 months had a significantly lower peak level of alanine aminotransferase (ALT)(718 ±696 vs 1282 ±913 U/L,622 ±514 vs 1203 ±924 U/L,P<0.05)and a significantly high-er level of HBV-DNA (6.8 ±1.4 vs 5.2 ±1.5 log10 copies/ml,7.3 ±1.6 vs 5.4 ±1.5 log10 copies/ml,P<0.05).The HBsAg clear-ance rate in patients with HBV genotype A was significantly lower than that in patients with HBV genotype B or D (75.0% vs 89.5% or 100%,83.3% vs 97.4% or 100%,P<0.05).Conclusion The HBsAg clearance rate in AHB patients may be associated with HBV genotypes,peak level of ALT,and HBV-DNA.

The traditional discipline-centered curriculum design can neither keep up with developments of modern medical science nor reach requirements of the education reform in the new century.Since 2011,College of Stomatology in School of Medicine in Shanghai Jiao Tong University had developed ‘ disease oriented integrated curriculum system reform’ for students of long-term stomatology education.In view of the problems existing in the original curriculum system,the integrated curriculum system was set up by coalescing clinical medicine curriculum according to the related systems and oral medicine curriculum according to the developmental rules of diseases.Lectures were combined with discussion classes in the reform and performance appraisal system was changed from simplex judgments into comprehensive evaluations.At last,further considerations of promoting the reform based on the practice were proposed.

Objective To investigate the satisfaction degree of students and teachers towards PBL teaching method and to provide a basis for continuing to carry out PBL teaching.Methods Anonymous questionnaires of PBL teaching method in teaching plan and procedure was conducted among 49students (2008 grade) and teachers.SPSS 13.0 software package was used to analyze the data and P ＜ 0.05 stands for statistically significant differences.Results Both students and teachers were satisfied.Teachers' evaluation scores of 17 projects were all higher than 4 points and students' evaluation scores of 13 projects were all higher than 4 points; scores for the rest projects were all higher than 3 points.Conclusions PBL teaching method combined with stomatology is effective and measures should be taken to strengthen the management of PBL teaching in stomatology.

Objective To study the effect of caspase-3 on the primary cultured mice cortical neuron w ith herpes simplex virus type 1 (HSV-1) infection. Methods A spectrophotometer was used to de tect the absorbance units at 405 nm of the caspase-3 substrate,the chrom o phore p-nitroanilide(pNA ).Comparing with the normal group, virus group, s orbital group,and sorbital group with virus in the absorbance units of chromophore substrate.The changes in inner mitochondrial membrane p otential of the different group cells were analyzed with flow cytometry. Results The released chromphore was measured by determining the absorbance at 405 nm. N group is 0.394 2, V group is 0.240 8, NS group is 0.539 2, VS gr oup is 0.465 0,the control C1 group is 0.290 0, C2 group is 0.235 0 and all groups were analyzed by statistic (P

Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

The purpose of this study was to fabricate decelluarized valve scaffold modified with polyethylene glycol nanoparticles loaded with transforming growth factor-β1 (TGF-β1), by which to improve the extracellular matrix microenvironment for heart valve tissue engineering in vitro. Polyethylene glycol nanoparticles were obtained by an emulsion-crosslinking method, and their morphology was observed under a scanning electron microscope. Decelluarized valve scaffolds, prepared by using trypsinase and TritonX-100, were modified with nanoparticles by carbodiimide, and then TGF-β1 was loaded into them by adsorption. The TGF-β1 delivery of the fabricated scaffold was measured by asing enzyme-linked immunosorbent assay. Whether unseeded or reseeded with myofibroblast from rats, the morphologic, biochemical and biomechanical characteristics of hybrid scaffolds were tested and compared with decelluarized scaffolds under the same conditions. The enzyme-linked immunosorbent assay revealed a typical delivery of nanoparticles. The morphologic observations and biological data analysis indicated that fabricated scaffolds possessed advantageous biocompatibility and biomechanical property beyond decelluarized scaffolds. Altogether this study proved that it was feasible to fabricate the hybrid scaffold and effective to improve extracellular matrix microenvironment, which is beneficial for an application in heart valve tissue engineering.

BACKGROUND: Diabetes causes abnormal insulin like growth factor-1 (IGF-1) gene expression, which contributes to initiation and development of peripheral neuropathy.OBJECTIVE: To investigate the efficacy of a single dose of methylcabalamin on prevention of experimental diabetic neuropathy and the possible molecular mechanism of its involvement in IGF-1 gene expression.DESIGN: Completely randomized and controlled experiment.SETTING: Endocrinology Department of the First Affiliated Hospital of Nanjing Medical University.MATERIALS: The study was carried out in an Animal Center of Nanjing Medical University. Totally 80 male Sprague Dawley rats (sanitary degree)were randomly selected.METHODS: ① Totally 64 rats were chosen to be induced diabetic. They were injected intravenously with alloxan dissolved in saline solutions, at the dose of 240 mg/kg. ② Of 16 rats were chosen as normal control group who were injected intravenously with equivalent volume of saline solution. ③ Of 64 established diabetic rats were treated with daily subcutaneous injection of pork regular insulin in combination of protamine zinc insulin (2:1) then further divided into 2 groups as insulin-treatment diabetic control groups based on different blood glucose levels: group 1 with relatively better control of diabetes, group 2 with relatively worse control of diabetes, with 32 rats in each group. Totally 16 rats of each group were treated with methylcobalamin injection intramuscularly with 500 μg/kg body weight, thus correspondingly divided into insulin +methylcobalamin group 1 and insulin+methylcobalamin group 2. The remaining 16 rats of each group as respective insulin-treatment diabetic control groups were treated with equivalent volume of saline. ④ Initiate weight and end weight were measured at beginning of the experiment and after diabetic model was established. Glucose oxidase was used to detect glucose level. 1-deoxy-1-malin was used to detect fructose level. ⑤ Parameters were measured as follows: Sensory/motor nerve conduction velocity (SNCV, MNCV) and evoked potential amplitude (EPA) of sciatic nerves detected by evoked electromyogram; IGF-I mRNA by reverse-transcriptase polymerase chain reaction (RT-PCR); IGF-1 peptide by enzyme-linked immunosorbent assay (ELISA). ⑥ One-way analysis of variance was used to analyze the Significance of differences among groups.MAIN OUTCOME MEASURES: ① Tissue IGF-1 mRNA/ IGF-1 peptide, electrophysiological data of individual groups at different points of the experiment. ② Comparison between individual groups in glucose metabolic parameters and body weights at different points of the experiment.RESULTS: Three rats died for diabetic infection or other acute complications and only 77 rats were included in the final statistical analysis.① Body weight and glucose metabolic parameter changes: After diabetic model, glucose, fructose level and body weight change between methylcobalamin+insulin treated groups and insulin treated groups were not significant. ② IGF-1 mRNA/peptide changes: Tissue IGF-1 mRNA increased significantly in methylcobalamin + insulin treated groups than that in insulin treated groups, respectively (P ＜ 0.05-0.01). Two weeks after diabetic model was established, the sciatic tissue IGF-1 mRNA contents were obviously higher in methylcobalamin + insulin treated group 1 than that in insulin treated group 1 (P ＜ 0.05), but not significantly different from that in NC group; Similarly, tissue IGF-1 mRNA contents were obviously higher in methylcobalamin + insulin treated group 2 than that in insulin treated group 2 (P ＜ 0.05), but lower than that in NC group (P ＜ 0.01); Month 2, tissue IGF-1 contents in methylcobalamin+ insulin treated groups were lower signiiicantly than NC groups, but higher than insulin treated groups (P ＜ 0.05-0.01). By month 3, IGF-1 mRNA level in methylcobalamin+ insulin treated group 2 was not significantly different from that in insulin treated group 2. The IGF-1 peptide levels in nerve tissue changed approximately parallel to IGF-1 mRNA level over time course. ③ Nerve electrophysiological data changes: Month 2 and 3, SNCV, MNCV and EPA were significantly higher in methylcobal-amin+ insulin treated group 1 than in insulin treated group 1 (P ＜ 0.05);Month 2, SNCV and EPA were higher in methylcobalamin+ insulin treated group 2 than in insulin treated group 2 (P ＜ 0.05); Month 3, SNCV, MNCV and EPA were significantly lower in methylcobalamin + insulin treated group 2 than in control group (P ＜ 0.05-0.01), whereas no difference was observed between methylcobalamin + insulin treated group 2 and insulin treated group 2.CONCLUSION: ① Methylcobal has not effect on blood glucose. ②Methylcobal could prevent occurrence of experimental neuropathy through its effect on nerve IGF-1 gene expression of diabetic rats. ③ A better efficacy could be achieved by Methylcobal with a good control of blood glucose level in prevention of diabetic peripheral neuropathy.

Objective To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the generation of human myeloid derived suppressor cells (MDSCs) relied on peripheral blood monocytes,and to establish efficient induction system in vitro of MDSCs.Methods Kidney transplantation recipients between January and March 2017 were included in this study.Purified CD14 + cells isolated from peripheral blood were cultured in the presence of GM-CSF with different concentrations for 7 days.Phenotypes and immunosuppressive abilities of induced MDSCs (iMDSCs) were investigated with FACS analyses.Inducible nitric oxide synthase (iNOS) mRNA expression was detected by qRT-PCR to determine the influence of iNOS-pathway on the immunosuppressive abilities of iMDSCs.Results A total of 11 recipients were included in this study.HLA-DR expression decreased sharply after the culture with GM-CSF.iMDSCs showed the similar phenotype characteristics with monocytic-MDSCs (M-MDSCs) as well as significant ability to suppress T cells proliferation and cytokines production.iMDSCs expressed higher levels of iNOS than monocytes,and the inhibitor effects of iMDSCs were significantly reduced after treatment with L-NMMA (1 mmol/L).The variations of phenotype and suppressive ability were concentrationdependent,and more significant changes could be revealed in the group of 10 μg/L GM-CSF.Conclusion GM-CSF-treated peripheral blood monocytes can be efficiently transformed to M-MDSCs,and suppress T cells proliferation and cytokines secretion via iNOS-dependent pathway.These results may contribute to establish MDSCs induction system,which would provide a basis for the clinical application of MDSCs.

Objective To discuss the methods of the heat preservation performance monitoring of the blood transport case and to provide the technical support for the safety of blood transportation.Methods At the different environment temperature,the amount of the cold resource was decided by the mass ratio of cold resource to blood and the temperature was automatically recorded by the intelligent temperature chip continuously,to monitor the changes of each monitoring point in the blood transport case.Results When the mass ratio of cold resource to blood was fixed at 1∶6,the cold chain of the blood transport case could keep the tempera-ture of 2-10 ℃ for 8 hours at the environment temperature of 12 ℃,It could keep the temperature of 2-10 ℃ for 4.5 hours at the environment temperature of 25 ℃,and it could keep the temperature of 2-10 ℃ for 2 hours at the environment temperature of 44℃.Conclusion When the mass ratio of cold resource to blood is fixed,as the environment temperature changes,the available time that the blood transport case keeps with the cold-chain requirement varies according to the results of the heat preservation per-formance monitoring of the blood transport case.

Objective:To study the formula of triptolide (TRI) self-microemulsifying drug delivery system (SMEDDS) and evaluate the pharmaceutical properties.Methods:The formula and preparation process of triptolide self-microemulsion were screened by the solubility test and pseudo-ternary phase diagram.With the average particle size and self-microemulsifying time as the indices,the further formula optimization of triptolide self-microemulsion was carried out.The pharmaceutical properties of triptolide self-microemulsion were evaluated.Results:The optimal formula of TRI SMEDDS was as follows:the amount of medium chain triglycerides (MCT) was 20%,that of polyoxyethylene castor oil (EL-35) was 40%,and that of polyethylene glycol 400 (PEG-400) was 40% in the oil phase.The average particle size was 43.48 nm,and the time of self-microemulsification was less than 30 s.Conclusion:The average particle size and the appearance of triptolide self-microemulsion are accordance with the requirements of pharmaceutics.Triptolide self-microemulsion has good sustained-release effect,which lays foundation for the further study on pharmacodynamics.