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.

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.

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.

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.

Objective: To study the fasting blood glucose level and associated factors relationship in physical examination population in Beijing. Methods: Totally 38 462 individuals who received physical examination from April 2006 through December 2007 in Peking Union Medical College Hospital physical examination center were enrolled in our study.We divided them into 8 groups according to the age,and the level of fasting blood glucose was tested.The relationship between BMI,age,sex and abnormity of blood glucose were analysed. Results: The incidences of overweight and obesity were 36.4% and 13.5%,respectively.The incidences of impaired fasting glucose and diabetes mellitus were 3.2% and 3.8%,respectively.The rate of abnormal glucose level increased with age and BMI.Compared to male,female persons had lower fasting blood glucose level and lower rate of abnormality(P =0.000).Compared to normal and thin population,overweight and obesity population had higher rate of fasting glucose abnormality(P

ObjectiveDiabetes is an independent risk factor for coronary artery bypasss grafting(CABG). Off pump coronary artery bypasss (OPCAB) experience in 251 cases was reviewed to determine whether diabetes wou ld be applicable in OPCAB procedures.MethodsConsecutive 251 patients underwent OPCAB over 12 month period. This study included 71 diebetic patients (DM group) and 180 nondiabetic patients (NDM group). Preoperative v ariables were compared between the two groups by univariate analysis.R esultsNo differences were found regarding the length of stay in cardio intensive care unit [DM group(2.4?0.3)d; NDM group (2.4?0.3) d;P=0. 386], and sternal complication (DM group: 5.7%;NDM group: 3.9%;P=0.511) . In hospital complications were as follows: death rate(DM group: 2.8%; NDM gr oup: 1.1%; P=0.680); stroke (DM group: 2 8%; NDM group: 1 7%; P=0 623 ); hemofiltratioin renal failure (DM group: 2.8%; NDM group: 0.5%; P=0.194); myocardial infarction(DM group: 0%; NDM group: 0.5%;P=1.000); blood using were more frequent in DM group comparied with NDM group (P=0.111). ConclusionOPCAB in diabetic patients is as safe as in non diabetic patients.

OBJECTIVE:To establish a method for simultaneous determination of spironolactone and erythromycin in Com-pound spironolactone gel. METHODS:HPLC was performed on the column of Thermo-Hypersil ODS2-C18 with mobile phase of 0.1 mol/L ammonium dihydrogen phosphate solution (pH was adjusted to 7.0 by triethylamine)-acetonitrile (60∶40,V/V) at flow rate of 1.0 ml/min,detection wavelength was 215 nm and 238 nm,column temperature was 30℃,and injection volume was 5 μl. RESULTS:The linear range was 0.251 6-5.032 μg/ml for spironolactone and 0.577 2-11.544 μg/ml(r=0.999 9) for erythromycin (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 0.83%;average recoveries were 97.8%(RSD=0.74%,n=9)and 96.7%(RSD=2.60%,n=9),respectively. CONCLUSIONS:The method is simple,reproducible,ac-curate and reliable,and can be used for the quality control of Compound spironolactone gel.

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.