Effects of RNA binding protein RNPC1 on biological function of renal cell carcinoma cells / 医学研究生学报

Objective RNPC1 may act as an oncogene or suppressor gene in human tumors and its role in human renal cell carcinoma (RCC) remains unclear.The objective of this study was to investigate the role of RNPC1 in the development of RCC.Methods Over-expression of RNPC1 gene group (RNPC1 group) and short hairpin RNA interfering RNPC1 gene expression (shRNPC1 group) were respectively built in RCC CAKI-1 and CAKI-2.The blank control group (NC group) and negative control group (SCR group) were built as well.The qRT-PCR and western blot (WB) were used to detect the expression levels of RNPC1 mRNA and RNPC1 protein in RCC cells.Lentivirus infection was applied to establish stable expressed RCC cell lines of RNPC1 over-expression and interference.Detection was made on mRNA and protein expression levels in RNPC1 stable RCC cell lines.The effects of RNPC1 on cell proliferation, colony formation assay, migration, and invasion were detected by CCK-8 cell differentiation test, clone test, scratch test, and migration and invasion test.WB was applied to detect the change of protein expression in the EMT path of RNPC1 stable RCC cell lines and explore the molecular mechanism of RNPC1 effect on the biological function of RCC cells.Results The expression levels of RNPC1 mRNA and protein were found lower in shRNPC1 group than those in SCR group, while the expression levels of RNPC1 mRNA and protein in SCR group were higher than those NC group (P<0.05).The capability of proliferation in shRNPC1 group was stronger than that in SCR group, while the capability of proliferation in shRNPC1 group was weaker than that in NC group (P<0.05).The capabilities of cell migration and invasion were stronger in shRNPC1 group than those in SCR group, while the capabilities of cell migration and invasion in RNPC1 group were weaker than those in NC group (P<0.05).RNPC1 could inhibit the proliferation capability of RCC cells and might up-regulate the protein expression of E-cadherin and down-regulate the protein expression of β-catenin and vimentin, thus inhibiting EMT path and the capabilities of migration and invasion off RCC cells (P<0.05).Conclusion RNPC1 acts as a tumor suppressor in RCC and has the potential for the prediction of RCC prognosis.

Modulation of TNF on the expression of protease activated receptors in P815 mast mells / 中华微生物学和免疫学杂志

Objective To investigate the potential influence of TNF on the expression of protease activated receptor (PAR)-1,2,3 and 4 by using P815 mast cells. Methods After being challenged with various concentrations of TNF for 2 h, 6 h and 16 h, the P815 mast cells were treated with or without Triton X-100 and the PAR expressions were detected by flow cytometry and immunofluorescent microscopy. Results Compared with the corresponding controls, TNF concentration-dependently upregulated expressions of PAR-2 and PAR-4 both in Triton X-100-treated and the untreated groups, but had no significant effect on the expression of PAR-1 and PAR-3. Moreover, no significantly different expressions of TNF-induced PAR-1, 2, 3 were observed between Triton X-100-treated and the untreated groups, whereas Triton X-100-treated PAR-4 expressions were significantly enhanced by TNF compared with the Triton X-100-untreated ones. Conclusion TNF can up-regulate PAR-2, 4 expression of P815 mast cells but has little effect on the expression of PAR-1, 3 correspondingly. And Triton X-100 treatment had no significant effect on TNF-modulated expression of PARs in P815 mast cells.

Identification and Antimicrobial Resistance of Mycoplasma from Female Cervical Secretions:Analysis of 2263 Cases / 中华医院感染学杂志

OBJECTIVE To study infection status and drug resistance of Ureaplasma urealyticum(Uu) and Mycoplasma hominis(Mh) in the female genital tract.METHODS The retrospective analysis of identification and the antimicrobial susceptibility testing of mycoplasma in 2263 female cervical secretions from Jan 2007 to Jun 2008 were conducted.RESULTS In 2263 cases,the positive rate of mycoplasma was 66.46%.The infection only by Uu accounted for 45.03%,by Mh for 1.46%,and the mixed infection for 19.97%.The results of drug sensibility test showed that drug resistances of mycoplasma were diffirent among three types of mycoplasma infections.CONCLUSIONS The infectious rate of mycoplasma from female cervical secretions is on big rise.It is important to culture and test the drug sensitivities of mycoplasma for choosing drugs rationally and control the resistant strains.

Antimicrobial Resistance among Clinical Streptococcus pneumoniae Isolates from Children / 中华医院感染学杂志

OBJECTIVE To investigate conditions and features of antimicrobial resistance in Streptococcus pneumoniae clincal isolates.METHODS Totally 203 strains of S.pneumoniae were isolated from Jan 2005 to Dec 2006 in Yuying Pediatric Hospital of Wenzhou Medical College.Then the antimicrobial susceptibility test to ten drugs and the MICs of penicillin were detected.RESULTS In 203 S.pneumoniae strains,all were sensitive to vancomycin,were sensitive to 98.0%,levofloxacin,the sensitivity of penicillin,erythromycin,tetracycline,clindamycin and SMZ was lower than 15.0%.The penicillin insensitivity of 203 strains reached 86.2%.There were 24 of the 203 strains for which penicillin MICs were ≥4 ?g/ml.CONCLUSIONS S.pneumoniae clinical strains are resistant to most of the commonly used antibiotics.The percentage of high level multidrug resistant strains is very high.Vancomycin and levofloxacin are the most sensitive drugs for S.pneumoniae clinical strains,the next are chlorophenicol.Erythromycin,SMZ,tetracycline and clindamycin aren′t useful in treating S.pneumoniae infection.

Identification and Drug Resistance of Mycoplasma from Semen of infertility Men / 中华医院感染学杂志

OBJECTIVE To study infection and drug resistance of mycoplasma from semen of infertility men. METHODS Mycoplasma from semen of infertility men was identified by cultivation,and the sensitivities to drugs were also performed. RESULTS In 267 cases the positive rate of mycoplasma was 46.07%.Simple infection of Ureaplasma urealyticum(Uu) accounted for 41.57%(11),and Mycoplasma hominis(Mh) 1.50%(4),and the mixed 3.00%(8).The result of drug sensitive test showed that sensitivities of mycoplasma to minocycline,doxycycline and josamycin were the highest,and then were roxithromycin and azithromycin.The drug resistance of mycoplasma to ofloxacin and clindamycin was the highest. CONCLUSIONS The infectious rate of mycoplasma from semen of infertility men is on big rise.It is important to culture and test the drug sensitivities of mycoplasma to use drugs rationally.

The apoptosis of HUVCE cell line induced by L-amino acid oxidase from snake of bungarus fasciatus / 中国病理生理杂志

AIM: To investigate the effect of purifried L-amino acid oxidase (LAO) from bungarus fasciatus snake venom on apoptosis and growth of HUVCE cell line. METHODS: The L-amino acid oxidase was purified by SP-sepharose HP column followed by Heperin-Sepharose (FF) column. The homogeneity of the preparation was examined by SDS-PAGE and the molecular weight of LAO was determined by SDS-PAGE and high performance liquid chromatography (HPLC) gel-filtration. The MTT assay was used to detect the viability of cells. Flow cytometry and laser confocal microscopy were used to identiyfy the cell cycle and apoptotic morphology after cells treated with LAO. RESULTS: An L-amino acid oxidase (BF-LAO) was successfully purified from the venom of bungarus fasciatus. It showed a single band in SDS-PAGE under both reduced and non-reduced conditions. The apparent molecular weight was determined to be 60 kD by SDS-PAGE and 70 kD by HPLC gel filtration. LAO inhibited growth and induced apoptosis of HUVCE cell line in a dose-dependent manner after 12 h incubation, with the 50% inhibitory concentration (IC_ 50 ) being of 2.8 mg/L. Flow cytometry and laser confocal microscope showed a typical apoptotic peak and morphological changes of these cells. CONCLUSION: The L-amino acid oxidase from bungarus fasciatus snake venom could inhibit the HUEVC cell growth and induce the cell apoptosis.