Objective To investigate the reversal effect of triazole antifungal itraconazole,fluconazole combined with adriamycin on multidrug resistance in leukemia cells.Methods Human chronic myelogenous leukemia adriamycin resistant cell lines K562/ADR cells were incubated with itraconazole,fluconazole,or PSC833 (positive control) combined respectively with adriamycin.CCK-8 assay was used to assess cell proliferation of K562/ADR.The mean fluorescence intensity of intracellular ADR was measured by flow cytometry.The marker of DNA damage γH2AX was detected by Western blot.Results 1 μg/ml itraconazole and 0.5 μg/ml PSC833 can decrease K562/ADR IC50 of adriamycin from 38.30 μg/ml to 8.59 μg/ml and 24.64 μg/ml in a dose-dependent manner.K562/ADR cells were incubated with 1 μg/ml itraconazole or 0.5 μg/ml PSC833 combined respectively with adriamycin for 3 h and 6 h,the mean fluorescence intensity of intracellular ADR were increased 1.54-fold (3 h),1.50-fold (6 h) or 5.97-fold (3 h),5.83-fold (6 h).Itraconazole or PSC833 combined with adriamycin significantly increase the expression of γH2AX in K562/ ADR cells.Conclusion Itraconazole can recover adriamycin sensitivity of K562/ADR by increasing the concentration of intracelullar adriamycin and synergistically increasing DNA damage,but not for fluconazole.

The surveillance of schistosomiasis in three sites of Mianzhu City after earthquake showed that there were no infected Oncomelania snails and cases,but the emerging area with snails were 7 895 m~2.Therefore,the control measures should be strengthened.

Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.

AIM:To investigate the effects of down-regulation of protein kinase C (PKC) on the activity of storeoperated Ca2 + channels (SOC) and the proliferation of airway smooth muscle cells (ASMCs). METHODS:Rat bronchial smooth muscle cells were isolated and cultured. Fluo-3 /AM fluorescence was measured by laser confocal microscope to assessing intracellular Ca2 +. Downregulation of PKC activity was achieved by incubation of ASMCs with PKC activator phorbol-12-myristate-13-acetate (PMA,10 ?mol/L) or phorbol 12,13 -dibutyrate (PDBu,1 ?mol/L) for 24 h. The proliferation of ASMCs was assayed by calculating the reduction rates of Alamar blue. RESULTS:Down-regulation of PKC activity by longterm exposure of PMA or PDBu inhibited the proliferation of ASMCs,the similar results were obtained by using PKC inhibitor chelerythrine. Both downregulation of PKC activity and inhibition of PKC activity by chelerythrine reduced Ca2 + entry through SOC channels. Low concentration of PMA (0. 1 ?mol/L) promoted the proliferation of ASMCs,and this effect was inhibited by SOC blocker SKF-96365. CONCLUSION:Inhibition or down -regu-lation of PKC activity results in the inhibition of SOC channels,suggesting that PKC is involved in the activation of these channels. Ca2 + entry through SOC channels might contribute to PKC-promoted proliferation of ASMCs.

AIM:To investigate the expression and function of store-operated calcium channels ( SOCC) in human circulating fibrocytes.METHODS:Peripheral blood mononuclear cells ( PBMCs) were isolated and cultured in ser-um-free media.After 7 d, the PBMCs differentiated into fibrocytes.RT-PCR and real-time PCR were performed to deter-mine the mRNA expression of ORAI1-3 and STIM1-2 in the fibrocytes.SOCC inhibitor SKF-96365 was used to elucidate the role of SOCC in the differentiation of fibrocytes.RESULTS:The results of real-time PCR showed that the mRNA ex-pression of ORAI1-3 and STIM1-2 was positive in cultured fibrocytes.SKF-96365 (10μmol/L) significantly inhibited the differentiation of fibrocytes.CONCLUSION:SOCC-related proteins ORAI1-3 and STIM1-2 are abundantly expressed in the fibrocytes, and may play an important role in the differentiation of these cells.

Objective To evaluate the clinical diagnosis and treatment of gastrointestinal stromal tumors(GISTs). MethodsThe clinical data of patients with GISTs admitted into surgical department from Jan 1999 to Mar 2004 were retrospectively analyzed. ResultsThere were 40 cases enrolled, with a male to female ratio of 30∶10. Thirty -eight patients underwent complete tumor resection, two did laparotomy only. According to the Fletcher classification, six patients were of very low risk, seven of low risk, eleven of intermediate risk, and sixteen of high risk.CD119 staining was positive in 80% cases, and CD34 was in 85%.Thirty-one patients were followed up, with 5 out of 29 postoperative patients suffering from recurrence and metastasis 6 to 30 months postoperatively. Two patients with unresectable tumor survived an average of 9 months. Gleevec was administered to two patients with partial relief achieved in one. Conclusion 1.GISTs are predominantly found in the middle-aged. Stomach and small bowel are most often affected. 2.Immunohistochemical staining was necessary for the diagnosis of GISTs.3.Fletcher classification is useful guide for clinical treatment and prognosis prediction.

To evaluate the efficacy of manipulation therapy in repairing thin-layer and thick-layer articular cartilage defects in rabbits.

Objective To assess the applicability of the Modification of Diet in Renal Disease (MDRD)formula to kidney function impaired Chinese diabetic patients.Methods Glomerular filtration rates(GFRs)in 463 Chinese diabetic patients(219 female,244 male,aged 14 to 88)were estimated by measuring ~(99m)Tc-DTPA clearance and with equations based on serum creatinine(Scr)and cystatin C(Cys C)concentrations.GFRs derived from various equations were compared with the ~(99m)Tc-DTPA clearance GFRs and their relative accuracies were assessed with ROC analysis.All the Scr measurements were performed with both the Roche enzymatic assay and the Beckman LX20 kinetic alkaline picrate assay,and Cys C with immunonephelometric and immunoturbidimetric assays.Results The reciprocals of Cys C and Scr were linearly correlated with ~(99m)Tc-DTPA clearance GFRs(r=0.830 and 0.690,repectively).The correlation of GFR with Scr could be expressed by an adjusted MDRD equation:GFR [ ml?(min?1.73 m~2)~(-1)]=175?(Scr)~(-1.154)?(age)~(-0.203)?0.742(female)?0.827,where 0.827 was a coefficient for Chinese.The adjusted equation showed a better accuracy than the MDRD equation(areas under the ROC curve 0.818 vs 0.644).The adjusted equation was also more accurate than equations obtained in previous Chinese studies.GFRs were also estimated by using Cys(in mg/L)with the following equation:GFR [ ml?(min?1.73 m~2)~(-1)] = 63.24?(Cys C)~(-0.3378).The accuracy of the Cys equation was similar to the Scr equation,or better in patients aged 60 and above.The Roche enzymatic results which were traceable to the isotope dilution mass spectrometry(IDMS)methods were significantly lower than Beckman LX20 results,but the results were closely correlated with each other(Y = 0.94X-0.02).When non-traceable Scr results were used,the coefficient needed to be adjusted.Conclusions GFRs can be estimated with equations based on either Scr or Cys C.GFR estimation should use standardized Scr results and take into account ethnic effects.

Objective: To investigate the inhibitory effect of adenovirus-mediated VEGF-siRNA on transplanted osteo- sarcoma in nude mice.Methods: VEGF-siRNA gene was cloned into the genome of replication-deficient adenovirus to construct Ad-VEGF-siRNA;the latter was then used to infect osteosarcoma MG63 cell line in vitro;and the expression of VEGF gene was detected by RT-PCR.Osteosarcoma transplantation model was established in nude mice;VEGF expression in tumor tissue was analyzed and the inhibitory effect on tumor growth and lung metastasis were also observed.Results: The recombinant adenovirus vector Ad-VEGF-siRNA was successfully constructed.In vivo and in vitro experiment both showed that Ad-VEGF-siRNA significantly downregulated VEGF expression in MG63 cells and transplanted tumor tissue. It was found that Ad-VEGF-siRNA significantly inhibited transplanted osteosarcoma growth(P

Objective To investigate the prognostic value of C5L2 in patients with hepatocellular carcinoma (HCC).Methods The data of 175 patients with HCC who underwent curative resection at Zhongshan Hospital,Fudan University from Oct.,2012 to Sep.,2013 were analyzed retrospectively.The correlation between C5L2 and clinicopathologic characteristics were explored.COX regression model was used to determine the influence of clinical parameters on predicting recurrence,and the prognostic value of combined application of C5L2 and AFP were evaluated by Kaplan-Meier method.In vitro,the expression of C5L2 were tested in 5 HCC cell lines,and Hep3B and Huh7 were chosen for down-regulation and up-regulation of C5L2,respectively,the abilities of invasion and migration were examined by transwell and the potential mechanism was explored.Results The C5L2 expression was correlated to gender,tumor size and recurrence,and the recurrence rate of low C5L2 expression group was higher.Also,the multivariate analysis showed that C5L2 low expression was an independent risk factor for recurrence.Moreover,the combined application of C5L2 and AFP could estimate prognosis more effectively.Knockdown of C5L2 in Hep3B promoted the invasiveness and motility,and increased the level of β-catenin and MMP2;conversely,overexpression of C5L2 in Huh7 inhibited the invasiveness and motility,and decreased the level of β catenin and MMP2.Conclusions C5L2 could be regarded as an auxiliary indicator for prognosis of HCC,thereby the evaluation of C5L2 could help with making effective and comprehensive management for HCC patients.