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 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 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.

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：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 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 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.

Objective To investigate the expression and significance of caveolin-1 in breast carcinoma.Methods Using immunohistochemical method the protein expression of caveolin-1 were analyzed in 105 cases of breast carcinoma and 50 cases of non-cancerous breast tissues.The relationship between caveolin-1 expression and CK5/6,EGFR,and E-cadherin expression was investigated.Clinical data of 105 cases of breast carcinoma were retrospectively analyzed.Results Among 105 cases of breast carcinoma,there were 20 cases of basal-like subtype,22 cases of luminal subtype A,23 cases of luminal subtype B,23 cases of HER2 over-expressing subtype,17 cases of normal breast-like subtype.Positive rate of caveolin-1 was significantly lower in breast carcinoma than in non-cancerous breast tissues (24.8％ vs.88.0％,P ＜ 0.05).Positive rate of caveolin-1 (75.0％) was higher in breast carcinoma than in luminal subtype A ( 4.8％ ),luminal subtype B ( 17.4％ ),HER2 over-expressing subtype ( 17.4％ ) or normal breast-like subtype( 11.8％ ),all P ＜0.05.Caveolin-1 expression was associated with expression of CK5/6 and EGFR(P ＜0.01 ).In univariate analysis,positive caveolin-1 was associated with higher lymph node metastasis rate (18/26,69.2％ )than negative (37/79,46.8％ ),P =0.047,and shorter 5-year-disease-free survival (38.46％ vs.74.68％,P =0.0004 ),but in multivariate analysis caveolin-1 was not an independent predictor of 5-year- disease-free survival (P ＞ 0.05).Conclusions Caveolin-1 can be seen as a screening mark of basal-like breast carcinoma,it may promote the invasiveness of breast cancer cells,but it is not an independent prognostic predictor of breast cancer patients.

Objective To establish a method of quantiying trace α-galactosidase from Bacteroides fragilis in enzymatic conversion of blood group B to O red blood cells ( B-ECO RBCs) .Methods BALB/c mice were immunized with purified recombinant B.fragilisα-galactosidase ( the purity>90%) to prepare monoclonal antibodies.The ascites were prepared using hybridoma cell lines stably secreting antibody and purified by HiTrap rProtein A column.The antibody titer and spe-cificity were detected by ELISA and Western blotting, respectively.Purified monoclonal antibody and rabbit polyclonal an-tibody were applied to detect residual enzyme in B-ECO RBCs and the washing solution was analyzed by indirect ELISA. Results A high titer and purity antibody was obtained.Western blotting showed that the antibody specifically reacted with B.fragilisα-galactosidase.Moreover, indirect ELISA was sensitive enough to detect the minimal amount of residualα-gal-actosidase at the concentration of 1 ng/ml.After four repeat washing cycles with 1∶4 ( v/v) phosphate-buffered saline, the amount of residual enzyme in B-ECO RBCs was less than 10 ng/ml.Conclusion An effective method of detecting the min-imal amount of residual α-galactosidase in blood conversion is established for safety evaluation of universal RBCs prepara-tion by enzymatic treatment.