Objective To construct Hepatitis B Virus (HBV) Chinese-strain wild type and Lamivudine-resistant variant recombinant baculovirus. Methods 1.2-unit length HBV construct (adr serotype, C genotype) was cloned into the multiple cloning region of the pFastBac Ⅰ vector and then transformed into DH10Bac, in which homologous recombination occurred between the pFastBac vector and the inside bacmid with the help of helper plasmid. So HBV recombinant bacmid was produced and transfected into sf9 cells to generate a Recombinant HBV baculovirus called vAcHBVc. The viral titers were detected by immune plaque assay and TCID 50. Lamivudine resistance mutation (YVDD) was introduced by site-directed mutagenesis using continuous PCR. HBV YVDD recombinant baculovirus (vAcHBVcYVDD) was generated and titered as above. Meanwhile, the recombinant HBV baculovirus containing GFP reporter was constructed for evaluating the efficacy of transfection and titration of virus. Results Recombinant wild and YMDD variant HBV baculovirus were successfully developed and the titer was between 10~6～10~8pfu/ml through plaque assay. Conclusion Different kinds of recombinant HBV baculovirus could be generated rapidly and efficiently with Bac to Bac expression system. The developed recombinant HBV baculovirus could be further used in the study of anti-viral resistances.

Objective To investigate the Shenmai injection for the inhibition of hepatic and renal toxicity and leukocyte disorder during chemoradiotherapy in the women with advanced breast cancer. Methods 58 cases of female breast cancer patients with stage Ⅳ were selected and randomly divided into 2 groups, 29 cases of each group, and patients were treated with 5-hydroxytryptamine (5-HT3) receptor antagonists and white blood cell growth hormone and other conventional therapy, the control group received gemcitabine plus cisplatin chemotherapy, 28d for 1 cycles, the treatment group received more with Shenmai injection, interval was 15d, 2 groups were treated for 3 cycles. Levels of peripheral blood T lymphocyte subsets, cytokine levels and liver and kidney function, quality of life and clinical efficacy were compared. Results Compared with before treatment, levels of CD3+, CD4+ and CD4+/CD8+ in control group decreased (P<0.05), levels of CD3+, CD4+ and CD4+/CD8+ in treatment group increased (P<0.05), levels of CD8+ decreased(P<0.05), levels of IFN-γ, IL-2 and TNF-α increased(P<0.05), levels of IL-6 decreased(P<0.05), scores of KPS increased(P<0.05), scores of FACT-B decreased(P<0.05), levels of ALT, AST, BUN increased(P<0.05), and levels of CCr, WBC counts decreased(P<0.05), and compared with the control group, levels of CD3+ , CD4+ and CD4+/CD8+ in the treatment group were higher(P<0.05), levels of CD8+ were lower(P<0.05), levels of IFN-γ, IL-2 and TNF-α were higher(P<0.05), levels of IL-6 were lower(P<0.05), and the total efficiency was higher(P<0.05), levels of ALT, AST, BUN were lower (P<0.05), and levels of CCr, WBC counts were higher (P<0.05). After treatment, the efficacy of treatment group was higher than that of control group(Z=-2.142,P=0.032<0.05). Conclusion Shenmai injection can improve the efficacy of radiotherapy and chemotherapy in patients with advanced breast cancer, and it can effectively inhibit the liver and kidney damage and leukocyte disorder.

Objective To observe the clinical efficacy of acupuncture plus Chinese medicine iontophoresis in treating dry eye syndrome.Method Sixty-two patients with dry eye syndrome were randomized into a treatment group and a control group, 31 cases in each group. The treatment group was intervened by acupuncture plus Chinese medicine iontophoresis, while the control group was intervened by artificial tears. After 4-week treatment, the changes of tear secretion, tear film break-up time (BUT) and symptoms score were observed, and the clinical efficacies were compared.Result The total effective rate was 87.1% in the treatment group versus 61.3% in the control group, and the between-group difference was statistically significant (P<0.05). The total symptoms score, tear secretion and BUT were significantly changed after the treatment in both groups (P<0.05). After the treatment, the total symptoms score, tear secretion and BUT in the treatment group were significantly different from those in the control group (P<0.05).Conclusion Acupuncture plus Chinese medicine iontophoresis is an effective method in treating dry eye syndrome.

Objective To establish a determining method for the content of berberine hydrochloride in Coptis chinensis Franch.and Xianglian pills.Method The sample was extracted and developed,the berberine hydrochloride was determined by thin layer chromatograph-fluorescence of spectrophotometry with Benzen-ethylaceate-isopropanol-methanol-ammonia water(6:3:1.5:1.5:0.5) as the developing system and Ex=365 nm,Em=409 nm.Result The calibration curve was linear in the range of 0.473～ 3.784 ?g,r=0.999 3.The recovery rate in Coptis chinensis Franch.was 99.03%,RSD=1.86%,and in Xianglian pills was 97.10%,RSD=1.09%.Conclusion The method is simple and accurate.

Based on the anatomic data and fractal theory, the vascular tree model of forearms similar to actual vessel was established, and then the 3D heat transfer model of forearms based on the actual vessel anatomic structure was created. The model was analyzed and investigated with finite element method to explore the influential factors such as dimensional distribution of vessel, blood perfusion rate and external heat source for the 3D temperature field. The simulation results were consistent with the experimental data derived from Pennes. The model well reflects the regional difference of tissue temperature caused by the irregular distribution of vessel, and provides temperature guidance for tumor thermotherapy.

According to teaching outline and clinical training characteristic of psychophy-laxis speciality(including Psychiatry and Medicopsychology),we explored how to strengthen the clinical thinking ability and clinical skill training for clinical medicine students based on cultiva-tion of elementary knowledge and theory,and we also established a set of more appropriate item system of clinical skills assessment so as to improve the practice quality in psychiatry and medical psychology departments.

Early diagnosis and early intervention for cognitive impairment after ischemic stroke can delay disease progression and prevent the occurrence of dementia.This article reviews the advances in research on ischemic stroke caused each risk factor for cognitive impairment.

Nitric oxide (NO) plays an important pathophysiological role in osteoarthritis and cartilage metabolism. To determine the relationship between NO and the synthesis of type II collagen in cartilage, we measured levels of type II collagen by ELISA and procollagen (II) mRNA by RT-PCR in cultured lapine chondrocytes that were incubated with some kinds of reagents. 0.2mM sodium nitroprusside (SNP, a NO donor) can release high levels of NO, decreasing type II collagen, suppressing the expression of procollagen (II) mRNA (COL2A1).At the same time, chondrocytes showed a large increase in NO synthesis, a decrease in type II collagen and COL2A1 mRNA in response to 100u/ml IL-1. When 1mg/ml N-nitro-L-arginine methyl easter (NAME, an inhibitor of NO synthase) was mixed with IL-1, NO production was inhibited, the amounts of type II collagen recovered partially and COL2A1 mRNA recovered completely.These data indicate NO can inhibit type II collagen synthesis as IL-1 downstream molecule by suppressing procollagen (II) mRNA.

Objective To analyze the viral dynamics and clinical significance during the antiviral treatment by a mathematical model.Methods Six chronic hepatitis B patients were evaluated with a kinetic model(Neumann model)during dose of 0.5 mg/d oral entecavir.Blood samples were drawn for HBV DNA measurement at week 0,2,4,12,24.Non-linear modeling was used to fit individual patient data.Results The median effectiveness in blocking viral production was 99.970%(n=6).The median half-life of viral turn-over was 1.6 d(n=6).The median half-life of infected hepatocytes was 21.3 d(n=5).Compared with the other patients,the c(virions are cleared at a rate)、ε(effectiveness)、δ(infected cell are lost at a rate)value of one patient(eg.6)were all lower and the half-life of virus and infected cells were higher,and eg.6 developed viral break-through after 38 weeks of follow-up.Conclusions Viral load decay showed a biphasic pattern during entecavir therapy which can be described with a mathematical model.The model relates processes of viral infection and replication as well as drug efficacy to model parameters.It indicates the prediction of bio-mathematical model during antiviral treatment.

Objective Following the prognosis of patients with myasthenia gravis(MG) who received both immunosuppressants and thymectomy, to determine whether the sequence and timing of treatment have influences on outcomes of MG and also to select significant predictors of prognosis.Methods It was a retrospective study which reviewed 105 patients with MG who underwent thymectomy and immunosuppressants between Januay, 2000 and December, 2013.All patients were divided into two subgroups based on their order of treatment.Thirty two patients were called as immunosuppressants-first group, and 73 patients were called as thymectomy-first group.The ratios of ideal status of two groups were compared with use of the ChiSquare test.Predictors of ideal status were analyzed by logistic regression.Results (1) After 1 year, 2 years and 5 years of intervention, the proportion of ideal status was significantly higher in the thymectomy-first group than that in the immunosuppressants-first group[(33％-50％) vs.(10％-20％), P ＜0.05].For oMG patients, after 1 year, 2 years and 5 years of treatment, thymectomy-first group had a higher ideal status rates than immunosuppressants-first group[(25％-50％) vs.(11％-18％)].But no significant difference was noted in the rates of ideal status at each follow-up time point.(2)Multivariate logistic regression analysis showed that sequence of treatment (OR =0.154, 95 ％ CI: 0.046-0.518, P =0.002) and symptom-thymectomy interval(OR =0.903,95 ％ CI: 0.835-0.976, P =0.010) were the significant predictors of ideal status.Conclusion Thymectomy prior to immunosuppressants is associated with relatively good prognosis.It is suggested that patients with gMG, or adult patients with oMG who are resistant to drug, will be better to undergo thymecotomy as early as possible.