Objective To explore the expressions of stromal cell derived factor 1 α (SDF-1α) and cysteine-X-cysteine chemokine receptor 4 (CXCR4) in the peripheral blood of patients with systemic lupus erythematosus (SLE).Methods Forty hospitalized SLE patients were recruited and twenty healthy volunteers were enrolled as normal controls.The percentage of CD3+CD4+CXCR4+,CD3+CD8+CXCR4+ and the plasma concentration level of SDF-1α in the control group and SLE patients were detected by flow cytometry and ELISA.The relationship between SDF-1α/CXCR4 and SLEDAI was explored.Kruskal-Wallis H,Pearson's and Mann-Whitney U test were used for statistical analysis.Results the expression of SDF-1α [329 (127,539) pg/ml] and CXCR4 [CD3+CD4+CXCR4+:4.20(2.01,6.35)％,CD3+CD4+CXCR4+:2.70(1.68,4.20)％] were significantly elevated in SLE patients when compared to the normal controls (Z=-6.277,-5.707,-4.885,respectively,all P=0.000),and were significantly increased in highly active SLE patients than less active SLE (Z=-5.414,-5.256,-5.312,P＜0.01).The expression of SDF-1α and CXCR4 in the butterfly erythema group,anemia group and proteinuria group were significantly higher than the normal group (P＜0.05).Both SDF-1αand CXCR4 were positively correlated with SLEDAI (r=0.857,0.830,0.861,respectively,all P＜0.01).Conclusion The expressions of SDF-1α and CXCR4 increase significantly in the peripheral blood of SLE patients and there is close relationship between SDF-1α/CXCR4 and disease activity,organ damage.The results of this study suggestthat SDF-1α/CXCR4 may play an important role in the pathogenesis of SLE.

Objective To investigate the expression of survivin/Human leukocyte antigen class I ( HLA-Ⅰ) proteins and its physiological significance in clear cell renal cell carcinoma ( CCRCC ) . Methods Immunohistochemistry was used to analyze survivin/HLA-Ⅰ protein expression in 90 cases of CCRCC and 10 normal tissues to study relationships with clinical symptoms and disease prognosis . Resutl s The level of survivin protein expression was found to be significantly higher in CCRCC tissues 82.2%( 74/90) than in normal tissues( 0/10).However, the relative amount of HLA-Ⅰprotein in colorectal cancer tis-sue was also found to be significantly lower 67.8%(61/90) than in normal tissues 90%(9/10).Survivin expression was associated with tumor grade , stage,and lymph node metastasis ( P=0.000, P=0.016, and P=0.001, respectively ) .Conversely , lost HLA-Ⅰexpression did not have any associations with clinicopath-ological data (P>0.05).Survivin negative patients (25.0%, 4/16) had a higher tumor-free survival rate than patients (52.7%, 39/74)with survivin expression (P=0.037).Patients (27.6%, 8/29) with normal HLA-Ⅰlevels had a higher tumor-free survival rate than those ( 60.7%, 37/61) with reduced HLA-Ⅰlev-els (P=0.020).The univariate and multivariate analyses indicated that expression of survivin and HLA indi -vidually and in combination were independent predictors for CCRCC patient survival . Conclusions Over-expression of survivin but reduced HLA-Ⅰ expression is associated with CCRCC development and progres-sion.

Objective To explore the CCR7 expression status in bladder urothelial carcinoma (BUC) and the relationship between CCR7 expression and lymph node metastasis,and analyze the impact of CCR7 expression on prognosis.Methods The expression levels of CCR7 in 57 BUC tissues and 10 normal bladder tissues were estimated by immunohistochemistry technique,and the correlation between CCR7 with lymph node metastasis,tumor stage,grade,number,size,relapse or not,and patients' age/sex of BUC was analyzed.The influence factors of lymph node metastasis were tested,and so were the influence factors of prognosis.Results The positive rate of CCR7 expression among 57 patients was 82.5％ (47/57) (high expression rate was 45.6％),which was higher than that in normal bladder tissue (20.0％,all were low expression,P ＜ 0.05).The high expression rate of CCR7 in lymph node metastasis group was 68.2％ (15/22),higher than that in none lymph node metastasis group (31.4％,11/35,P ＜ 0.01).The expression level of CCR7 had no significant correlation with tumor stage,grade or other parameters.CCR7 expression,tumor stage and tumor grade were correlated with lymph node status (P ＜ 0.05),but only the first was an independent one.High CCR7 expression had a significant link with low relapse free survival (P ＜ 0.05).Conclusion The expression of CCR7 was highly expressed in BUC,which may be a positive independent influence factor of lymph node metastasis,and a predictor of poor prognosis.

Objective To evaluate the effect of health education in controlling the iodine deficiency diserders(IDD) in order to provide reference data for the further prevention and control. Methods Each village of 3 towns in Congjiang County was selected in 2007, where the health education lasting for 10 months had been implemented in the school students of 3-6 grade and the villagers. The school students of 3-6 grade and 30 housewives in the villagers were investigated for their IDD control knowledge, the salt consuming conditions as well as the sales of both rough and fine salt at a salt retail site in each village before and after the health education was implemented. Results The awareness rate of the knowledge of IDD control in the students and housewives was 91.4% (581/636) and 78.3% (282/360), respectively after intervention, which significantly increased (χ2= 532.044, 326.117, both P < 0.01) compared with the rate of 28.2% (184/652) and 11.4% (41/360) before intervention. The proportion of consuming fine salt was 91.8%(146/159) and 95.6%(86/90), significantly inereased(χ2= 236.623, 135.350, both P < 0.01) compared with 6.1%(10/163) and 7.8% (7/90) found before intervention. The selling proportion of fine salt at the salt retail site in the village was 60.0%(900/1500), significantly increased(χ2= 824.176, P < 0.01) compared with 10.0%(150/1500) before intervention. Conclusions Health education and promotion is solid foundation for effectively controlling IDD, through which the students and villagers are actively and voluntarily involved in the program and hence have formed good living and hygiene habits, thus expected effect has been obtained.

The technique conditions of decolonization of fermentation broth were successively optimized using single factor assay and orthogonal layout method in Cordyceps jiangxiensis. The optimal condition of decolorization was investigated to be 3g/100mL active carbon, 5 min absorption time, pH5 of fermented broth and 25℃absorption temperature. Under the optimal condition, the maximum decolorization rate of fermented broth reached 89. 6% , simultaneously 10. 7% consuming rate of exopolysaccahride was minimum. Subsequently, the extract condition of exopolysaccharide of C. jiangxiensis was further optimized by orthogonal layout design. The maximum exopolysaccharide production was 0. 38 g/L under the optimal condition, i. e. firstly fermented filtrate decolorized and deproteined was concentrated to 1/7 of its total volume, secondly concentration broth was mixed with four times its volume of absolute ethanol and stirred vigorously, lastly precipitation of exopolysaccharide proceeded at 4℃for 16 hrs and the exopolysaccharide collected by centrifugal ion and dryness.

To explore the plasticity of human amniotic mesenchymal cells(hAMCs) into cardiomyocyte-like cells,hAMCs were isolated from human amnion with collagenase digestion.Phenotype of the isolated cells was analyzed by flow cytometry(FCM).hAMCs were treated with 5-azacytidine and basic fibroblast growth factor(bFGF) to investigate their ability of differentiation into cardiomyocytes.The induced differentiated cells were evaluated by immunofluorescence for desmin and ?-actin expression and by RT-PCR for Nkx2.5,GATA-4 and alpha-myosin heavy chain(?-MHC) mRNA expression.The results showed that,after primary culture,hAMCs could reach a confluence of 80% with swirl like growth at 6 days.The cells proliferated rapidly after passages with a 100% confluence at 3~4 d.hAMCs were positive expression of CD44 and vimentin,but negative for CK19.After induced differentiation at 8~10d,the differentiated cells have close-up arranged with long spindle-shape.At 2 weeks and 4 weeks,induced cells expressed ?-actinin and cardiac-specific transcription factor Nkx2.5.Expressions of GATA-4 and desmin can be detected but ?-MHC can not in the hAMCs both before and after the induction.In conclusion,hAMCs have the ability of differentiation into cardiomyocyte-like cells,which means that hAMCs can be regarded as candidate cells for cellular cardiomyoplasty(CCM).

OBJECTIVETo study the antiproliferation effect on HepG2 cells and its underlying mechanism of the active chemical composition of the Viburnum Odoratissimum.

METHODS3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and trypan blue dye exclusion assay were used to assess the effect of vibsane-type diterpenoids on the proliferation of various tumor cells. Alterations in cell cycle and apoptosis were determined by flowcytometry. The enzymatic activity of caspase-3/7 was measured by Apo-ONE homogeneous Caspase-3/7 Assay kit.

RESULTSCompound 1 #, a vibsane-type diterpenoid, was found to significantly inhibit the growth of HepG2 cells by anticancer proliferation activity screening. It was demonstrated that the modified groups on side chain coupled to C11 site affected the cell growth-inhibition activity of compounds by structure-activity analysis. In addition, HepG2 cell line was most sensitive to compound 1 #, which induced growth arrest of HepG2 cells in a dose- and time-dependent manner. Study on the mechanisms underlying these effects indicated that compound 1 # induced significant G0/G1 phase arrest of HepG2 cells in a time- and concentration-dependent manner. Meanwhile, It was found that higher concentrations of compound (5-10 micromol/L) caused evident increase in the unmber of apoptotic cells and dose-dependent activation of caspase-3/7.

CONCLUSIONVibsane-type diterpenoids could significantly inhibit the growth of HCC HepG2 cells. Induction of cell cycle arrest and apoptosis may play important roles in their anticancer effects.

Apoptosis ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Diterpenes ; pharmacology ; Hep G2 Cells ; Humans ; Viburnum ; chemistry

Objective Take alkaline ashing method as golden standard to explore the accuracy of chloric acid digestion method in determination of human milk iodine. Methods Sixty one breast milk samples collected in Hexi district of Tianjin was measured by the method for determination of iodine in foodstuff by As3+-Ce4+ catalytic spectrophotometry (referred to as the alkaline ashing method) published in 2008 and the method for determination of iodine in urine by As3+-Ce4+ catalytic spectrophotometry(referred to as acid digestion) published in 1999, respectively. were highly correlated(r = 0.960, t = 26.3, P < 0.01), and the regression equation was (Y) = - 28.1 + 0.808X, in which X was independent variable, that is the results of alkaline ashing method; (Y) was dependent variable, that is the estimated data of chloric acid digestion method. The average difference of the results measured by the two methods was 68.3 μg/L, and the results from chloric acid digestion was 38.9% which lower than that of alkaline samples were diluted by 3,4 and 5-fold and then digested by chloric acid, the liquid clarification rates were 80.3% ashing and chloric acid digestion method were, respectively, 165.4, 110.0 μg/L. Conclusions Compared with alkaline ashing method, the results determined by chloric acid digestion method are significantly lower. It is suggested that there are systemic errors in chloric acid digestion method, which means that alkaline ashing method can not be replaced by the chloric acid digestion method.

To study in vitro anti-influenza viral activities of Chinese traditional patent medicines for influenza prevention and treatment, neuraminidase (NA) activity assay was used to examine NA inhibitory activity of 33 Chinese traditional patent medicines through fluorimetric assay, and influenza virus induced cytopathic effect (CPE) inhibition assay was used to verify their anti-influenza viral activities in vitro. The assay results showed that most liquid preparations displayed relatively high NA inhibitory activities, such as Shuanghuanglian oral liquid, Qingkailing oral liquid, Qingre Jiedu oral liquid, and Reduning injection. Among liquid preparations, Shuanghuanglian oral liquid not only displayed the highest NA inhibitory effect, but also exhibited obvious in vitro anti-viral activity in CPE experiment. Among solid preparations, Shuanghuanglian powder for injection showed the highest activity on NA inhibition, and Fufang Yuxingcao tablet showed relatively strong anti-influenza viral activity in CPE cells. From the results, it can be concluded that most Chinese traditional patent medicines possessed NA inhibitory activity, but only a few of them displayed significant in vitro anti-influenza viral activities. These results will provide important information for the isolation of active constituents, and for the clinical uses of Chinese traditional patent medicines for influenza treatment and prevention.
Animals ; Antiviral Agents ; pharmacology ; Cell Line ; Cytopathogenic Effect, Viral ; drug effects ; Dogs ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Influenza A Virus, H1N1 Subtype ; drug effects ; enzymology ; Influenza A Virus, H3N2 Subtype ; drug effects ; enzymology ; Influenza B virus ; drug effects ; enzymology ; Medicine, Chinese Traditional ; Neuraminidase ; antagonists & inhibitors ; metabolism ; Plants, Medicinal ; chemistry

OBJECTIVETo study inhibitory efficacy of combined gene therapy for malignant gliomas transfected with antisense human telomerase reverse transcriptase (hTERT)/PTEN in vitro and in vivo.

METHODSTo construct two adenovirus recons which contained antisense hTERT and wild-type PTEN respectively with high performance homologous recombination system in bacteria. The two adenovirus recons were transfected into U251 human malignant glioma cells combinedly or respectively in vitro and in vivo. U251 cell proliferation in vitro was determined by MTT assay and flow cytometry, tumor growth in vivo was measured by the volume of glioma in nude mice. Telomerase activity was detected by telomeric repeat amplification protocol (TRAP) assay. Expression of hTERT and PTEN protein was detected by Western blotting methods.

RESULTSAfter transfection in vitro, the growth of U251 cells was inhibited significantly. The inhibitory effect was time-dependent. The strongest inhibition was observed in combined transfection group, at the 6th day, the survival rate was 37.6%, telomerase activity (only 28.8TPG) was inhibited significantly, hTERT protein expression was inhibited significantly too, which was 0.2106, but PTEN protein expression was increased significantly, which was 0.9630. In vivo, the growth of tumors was also effectively inhibited.

CONCLUSIONGrowth of malignant glioma cells is effectively inhibited after transfection with combined antisense hTERT and PTEN in vitro and in vivo.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Blotting, Western ; Brain Neoplasms ; pathology ; therapy ; Cell Line, Tumor ; Cell Proliferation ; DNA, Antisense ; genetics ; metabolism ; Flow Cytometry ; Genetic Therapy ; methods ; Glioma ; pathology ; therapy ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Mice ; Mice, Nude ; Microscopy, Fluorescence ; PTEN Phosphohydrolase ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Telomerase ; genetics ; metabolism ; Transfection ; Tumor Burden ; Xenograft Model Antitumor Assays