AIM To investigate the inhibition and apoptosis mechanism of GBC-SD cells induced by rubescensine B. METHODS Using MTT, convert microscopy, electron microscopy, flow cytometry, an immunohistochemical assay, and spectrofluorometry demonstrate the presence and pathogenesis of apoptosis after treated by rubescensine B. RESULTS After exposure to Rubecensine B GBC-SD cells were induced to apoptosis in dose-dependent manner, and the level of Bcl-2,p53,C-myc,Fas/APO-1 were decreased within 24 hours, reversely the activity of Caspase-3 was enhanced with the appearance of apoptosis. CONCLUSION Rubecensine B can induce GBC-SD cells apoptosis related to Bcl-2,p53,Fas/APO-1 and C-myc.

Objective:To explore the genotype distribution of lipoprotein lipase polymorphism at Pvu Ⅱ locus and its possible association with serum lipid levels, body mass index (BMI) and dietary intake. Method:Pvu II polymorphism was determined by PCR-PFLP method in 156 adults with hyperlipidemia and 154 adults with normal serum lipid levels as control in Shanghai. Their physical examinations, dietary investigation and the levels of serum lipid profile, including TG, TC, HDL and LDL were analyzed. Results: ⑴ There was no significant difference in frequencies of genotypes or allele between the hyperlipidemia and the control, and between male and female. ⑵ BMI were significantly different between P-P-and P+P+, P+P-genotype groups. The BMI of subjects with genotype P+P+ had the highest levels. The relationship was significant after adjusting several confounding factors such as age, gender and blood lipid (P

Objective To examine clusters of anti-nuclear antibodies (ANA) and their associations with clinical features in patients with systemic lupus erythematosus (SLE).Methods It was a retrospective study.113 SLE patients were reviewed from March 2010 to May 2012 in Department of Rheumatology,Jinhua Central Hospital.ANA and specific autoantibodies to 15 kinds of nuclear antigens were tested by indirect immunofluorescence assay (IIF) and line immunoassay (LIA) respectively.Hierarchical clustering method was performed to analyze specific clusters of ANA profiles in SLE.Chi-square tests were used to investigate relationship between antibody clusters and clinical features of SLE.Results The positive rate of LIA for ANA was 97.3％,consistent with IIF method,and the total accordance rate of the both methods was 98.2％.Thirteen kinds of antigen-specific antibodies were detected in SLE patients by LIA.Clustering analysis for these antibodies showed three specific clusters in SLE,Nuc/His/dsDNA cluster (C1),low-Ro/low-La cluster (C2),and Ro/Sm/RNP cluster (C3),accounting for 36.3％,24.8％,38.9％ of the total cases respectively.There were significant difference of AST levels among three clusters [(32.62 ± 21.92)U/L,(25.56 ± 16.63) U/L,(50.41 ± 60.86) U/L respectively for C1,C2 and C3].High incidences of chronic cutaneous lupus,abnormal renal indicators and inflammatory synovitis were found in all three clusters.Besides,there were significant differences among three clusters for the incidences of chronic cutaneous lupus (39.0％,39.3％,63.6％ respectively for C1,C2,C3) and leukopenia/lymphopenia (56.1％,25.0％,56.8％ respectively for C1,C2,C3) (P ＜ 0.05).Patients in Ro/Sm/RNP cluster showed higher incidences of lupus nephritis (43.2％/26.8％ or 39.3％); patients in low-Ro/low-La cluster showed low risk of hypertension (7.1％/19.5％ or 22.7％) ; patients in Nuc/His/dsDNA cluster showed high incidences of thrombocytopenia (41.5％/21.4％ or 25.0％) and high risk of lung or upper respiratory tract infection (46.3％/28.6％ or 29.5％),but low incidence of neurologic symptoms (0％/ 3.6％ ％ or 11.4％).Conclusion Three characterized ANA clusters are identified in SLE patients in this pilot study.Different clusters are associated with certain clinical features and complications ofSLE.However,the correlations found in this study need to be investigated further in larger populations.

Objective：Our aim was to investigate the proportion of autosomal recessive (AR) inheritance among families with patients with Duchenne muscular dystrophy (DMD) and clinical feature in patients with AR form of DMD. Methods:A total of 193 families was studied， 8 of them with at least one girl with “DMD - like” phenotype and 185 with only boys with this kind of phenotype. Based on the number of families with at least one affected girl and the number of patients per sibship among these pedigrees, the proportion of families with DMD inherited as an AR trait was estimated. The clinical examination, family history and serum creatine-kinase were studied in 11 patients diagnosed as AR form of DMD. Results: The proportion of families with AR form of DMD was estimated as 9.4%. The average age of being able to walk is (1.47±1.00) year, serum creatine-kinase levels were (2785.10±1500.29) U/L. The clinical symptom occurred at the average age of (8.11±4.32) year in patients with AR form of DMD. Conclusion： The AR form of muscular dystrophy and DMD not be distingushed clinically. Some families with only affected boys diagnosed as typical DMD, in fact, have the AR form of the disease. This study is very useful for genetic consulting.

Objective To explore the genotype distribution of lipoprotein lipase gene polymorphism at Ser447Stop locus and its possible association with metabolic syndrome and dietary intakes. Method Ser447Stop polymorphism was determined by PCR-PFLP method in 222 adults with MS and 222 normal adults as control. Their physical examination,dietary investigation and levels of biochemical profile,including BG,TG,TC and HDL-C were analyzed. Results (1) The genotype frequencies of Ser447Stop SS,SX and XX were 85.6%,13.3% and 1.1% respectively,which were in agreement with Hardy-Weinberg equilibrium. There was no significant difference in frequencies of genotypes or allele between MS and the control,and between male and female. (2) After adjusting age and gender,the levels of serum TG were significantly different among three genotype groups,the highest in SS genotype and the lowest in XX genotype. (3) After adjusting age,gender and body mass index,the intakes of protein and carbohydrate were significantly different among three genotype groups. (4) There was significantly different in negative correlation between the intakes of protein and serum TG levels after adjusting age,gender and body mass index. Conclusion Lipoprotein lipase gene polymorphism influenced serum TG levels,while associated with protein intakes. It might contribute to the predisposition in metabolic syndrome response to dietary intervention.

OBJECTIVE: To evaluate the efficacy and toxicity of sorafenib plus Capecitabine in patients with advanced hepatocellular carcinoma (HCC). METHODS: 20 patients (treatment group) were assigned to take sorafenib 200 mg bid for 3 consecutive weeks plus capecitabine 1 500 mg? m-2?d-1 for l4 days followed by 7 days discontinuation in 3-week treatment cycle. 22 patients in the control group only received Capecitabine 1 500 mg?m-2?d-1 for l4 days followed by 7 days discontinuation in a 3-week treatment cycle. Tumor response was assessed after 2-cycle treatment using modified WHO criteria. RESULTS: In the treatment group and the control group: the median survival times were 10.9 months and 7.2 months, respectively; the median time for tumor progression was 6.8 months and 4.3 months, respectively; the overall response rates were 20.0% and 9.1% respectively; the clinical benefit rates were 70.0% and 40.9%; the ?-foetoprotein (AFP) reduction rates were 65.5% and 39.0%, respectively. The toxicities were not significant between the two groups. CONCLUSION: Sorafenib plus Capecitabine is safe and effective for advanced hepatocellular carcinoma patients.

Object To study the effects of artemisinin and its analogue artemisunate on the proliferation of human breast cancer MCF-7 cell line, as well as their mechanism comparatively. Methods The inhibition of cell proliferation was determined by SRB method. Cell cycle was determined by flow cytometry (FCM) analysis. Apoptosis was confirmed by sub-G 1 cells content and DAPI method. Results The cell cycle of MCF-7 was changed greatly when treated 24 h with either 10 ?mol/L artemisinin or 1 ?mol/L artemisunate, the distribution of MCF-7 cells among S phase was reduced greatly, while inereased during G 0+G 1. However, artemisinin had weaker effect on the proliferation of MCF-7 cell, while artemisunate effectively inhibited the proliferation of MCF-7, the IC 50 was 0.31 ?mol/L. Apoptosis induced by 1 ?mol/L artemisunate was stronger than that by 10 ?mol/L artemisinin, too. Conclusion The inhibitory effect of artemisunate on the proliferation of tumor cell is stronger than that of artemisinin in vitro.

Objective To observe reverse effect of anisodamine to the adverse effect of remifentanil during enteroscopy without pain for patients with bradycardia .Methods Sixty‐five patients with bradycardia were selected and divided randomly into group C (n=21 ,control group)、group A1 (n=22 ,anisodamine by instillation) and group A2 (n=22 ,anisodamine by continous infusion) .In‐duction :Intravenous etomidate 0 .08 mg/kg ,propofol 1 .00 mg/kg and remifentanil 0 .10μg/kg in 3 groups .Ten mg anisodamine in‐fused by instillation before induction in group A1 ,5 mg anisodamine infused by instillation before induction and continous infused by 0 .25 mg/min in group A2 .Maintenance:All group received propofol 4 mg · kg‐1 · h‐1 ,remifentanil 0 .05 μg · kg‐1 · min‐1 after un‐dergoing enteroscopy .stopping pumping propofol when colonoscopy reached ileocecal junction ,and we took off remifentanil when colonoscopy withdraw to decending colon .Then we observed and recorded HR ,SpO2 ,MAP ,dosage ,fluid infusion ,induction time , check time ,analepsia time ,degree and of enterospasm and numbers of cases and side effect at T0 (before induction) ,T1 (beginning of operation) ,T2 (into the transverse colon) ,T3 (to the ileocecal junction) ,T4 (exit) .Results There were no significant difference a‐mong 3 groups of induction time .Compared with group A1 and group A2 about check time and analepsia time ,group C was much shorter .The HR of group A1 and A2 were more stable than group C at T2 、T3 .At T1 、T2 ,the fluctuation of HR of group A2 was less than that of group A1 .There was obviously different among 3 groups of propofol′s dosage ,operation time and enterospasm ,the effect of group A1 and group A2 were better .There was statistically significant in number of cases of body movement between group A1 (1/22)and group C(4/21) ,there was also statistical significance between group A1 ,group A2 and group C(P<0 .05) .Conclusion There are no difference between 2 methods about relieving enterospasm ,refraining intestinal angina ,shortening operation time , saving anesthetic dosage .Effect of continous pumping to undulation of HR may be more stable .

Objective To investigate whether IL-17 could stimulate the vascular endothelial growth factor (VEGF) production on HaCaT cells alone. We also investigated whether shikonin could inhibited the proinflamation effects of interleukin-17(IL-17) acting on HaCaT cells. MethodsWe examined the expression of VEGF by double antibody sandwich enzyme-linked immunosorbent assay ( ELISA ) and realtime polymerase chain reaction(RT-PCR) in HaCaT cells and the cell supernatant. The viability of HaCaT cells in the drug group was detected by the Cell Counting Kit-8 (CCK-8). ResultsThe expression of VEGF in different time IL-17-stimulated groups on HaCaT cells and the cell supernatant were higher than the control group( P＜0.001 ). The expression of VEGF in different drug treatment groups on HaCaT cells and the cell supematant were lower than the stimulated group by IL-17 ( P＜0. 001 ). The cell viability of different drug treatment groups have no significant difference( P＞0.05 ). ConclusionWe show that IL-17 specifically and time-dependently augmented and induced VEGF expression on HaCaT cells and the cell supernatantThen shikonin markedly inhibited the increase tengency of IL-17 effection on HaCaT cells and the cell supematant level.

Objective To observe whether rabbit blood leptin can leak into the brain after craniocerebral trauma (CCT),and to dynamically detect serum levels of leptin,growth hormone (GH),and insulinlike growth factor-1 (IGF-1),and leptin level in cerebrospinal fluid (CSF) after CCT.Methods The FITC labeled leptin was injected into blood vessels of 15 adult male rabbits after anesthesia.Then 9 rabbits underwent unilateral fluid percussive impact,while 6 rabbits underwent sham operation.Thirty minutes postoperatively,brain tissue was taken to make frozen sections which were used to observe fluorescence labeled leptin range.Thirty three adult male rabbits were randomly divided into serum group,serum-control group,CSF group and CSF-control group.Rabbits in serum group and CSF group received fluid percussive impact,while rabbits in serum-control group and CSF-control group were drilled a 7 mm window in skull.Rabbits in serum group and serum-control group were phlebotomized 2 ml at 1 d,3 d,7 d and 14 d after operation,and rabbits in CSF group and CSF-control group were extracted CSF 0.5 ml at the same time points.Then serum levels of leptin,GH,and IGF-1,and leptin level in CSF were tested by ELISA.Results The fluorescence imaging could be seen in the injured brain tissue of rabbits with CCT,which was more than those in brain tissue of rabbits receiving sham operation.Serum leptin levels of rabbits in serum group at each time point were higher than those in serum-control group.Serum levels of GH and IGF-1 and leptin level in CSF were higher in rabbits with CCT than those in rabbits without CCT at 1 d,3 d and 7 d after operation.Conclusion The blood leptin can leak into the brain after CCT,which can cause increase of blood GH and IGF-1.And the latter may be the endocrine factors promoting fracture healing after CCT.