Objective The increasing evidences indicate that the inhalable particle matter (PM10) is responsible for respiratory and cardiovascular morbidity and mortality.Generation of superoxide anion,hydroxyl radical and apoptosis in HLF cells induced by PM10 were studied in this paper to present some data for elucidating the mechanism of effects of PM10.Methods The superoxide anion and hydroxyl radical were detected respectively by cytochrome C assay and apoptosis inspected with Annexin V-FITC/PI double staining by flow cytometry.Results PM10 could generate the superoxide anion and hydroxyl radical,with significant dose-dependent manners,and the levels of the superoxide anion as well as hydroxyl radical were decreased by chitosan oligosaccharides (COS),with the significant protection effects (P

Objective To investigate curative efficacy of RevoLix 2 μm laser in combined with pirarubicine in the treatment of non-muscular invasive bladder cancer and its effects on level of serum vascular endothelial growth factor (VEGF).Methods 115 cases with non-muscular invasive bladder cancer who received therapy in our hospital from January 2012 to April 2013 were selected as research objects to retrospectively analyze their materials.According to therapeutic schemes,patients were divided into group A ( n=52 ) and group B ( n =63 ).Group A was given electrocision of bladder tumor and perfusion of pirarubicine 30 mg in 5% glucose injection 30 mL,one times per week,for eight weeks,and then one times per month,a total of one year; while group B was given RevoLix 2μm laser resection of bladder tumor and perfusion of pirarubicine 30 mg in 5% glucose injection 30 mL,one times per week, for eight week, and then one times per month, a total of one year.Then, operation status, incidence of complications after operation,level of VEGF and recurrence rate between the two groups were observed and compared.Results Compared with group A, group B had statistically less bleeding volume,shorter retention time of urinary tube and hospitalization time (P<0.05),while operation time in the two groups was statistically same.After operation,incidence of complications in group B was 3.2%,which was statistically lower than that of 15.4% in group A ( P<0.05).On admission and 1 week after operation,levels of VEGF of two groups had no significant differences.8 weeks and 6 months after operation,level of VEGF in group B was statistically lower than that in group A respectively (P<0.05).After the two-year follow-up,recurrence rate in group B was 4.8%,which was statistically lower than that of 17.3% in group A (P<0.05).Conclusion RevoLix 2μm laser combined with pirarubicine is effective for patients with non-muscular invasive bladder cancer with less bleeding volume,quicker postoperative recovery,lower incidence of complications and recurrence rate after operation.This may be related to the decrease of serum VEGF level in patients,it is necessary to further study.

Objective To explore the relationship of signal transduction among human papillomavirus 18 E6 oncoprotein (HPV18E6), signal transducers and activators of transcription 1 (STAT1), protein kinase R( PKR )/α subunit of eukaryotic initiation factor 2 ( eIF2α ), nuclear factor-kappa Bp65 ( NF-κBp65 ), mitogen-activated protein kinase( MAPK)/c-Jun N-terminal kinase(JNK) ,and possible molecular mechanism. Methods Construct two lentiviral vectors which contain shRNA interfering sequence aiming at the targets of HPV18E6 oncogene and NC sequence( HPV18E6-RNAi-LV, NC-GFP-LV), based on the transduction with HPV18E6-RNAi-LV and NC-GFP-LV into HeLa cell to interfere the expression of HPV18E6 oncogene and NC sequence,the expressions of mRNA and protein( including phosphating patem)of HPV18E6, STATI, PKR, eIF2α, NF-κBp65, MAPK, JNK are measured with RT-PCR and Western blot, the difference of proliferation and sensitivity to carboplatin of HeLa cell are determined with Transwell cell methods and MTT among every groups. Results The expression of HPV18E6 oncogene can affect the expression level of mRNA and protein of NF-κBp65 and PKR genes, also affect phosphating levels of phosphating protein p-STAT1, p-PKR and p-eIF2α;the restraining rates of proliferation and sensitivity to carboplatin of HeLa cell are higher in HPV18E6-RNAi-LV group than the other groups( P＜0. 05 or P＜0.01 ). Conclusion HPV18E6 oncoprotein not only reduces the expression of PKR but dephosphorylates p-STAT1, pPKR and p-eIF2α to restrain activation of PKR/eIF2α signal transduction passage, maintain the proliferation and invading ability of HeLa cell and restrain apoptosis. The signal transduction among HPV18E6, MAPK/JNK are not clear.

Objective To isolate cancer stem cells from human colon cancer cell line CW-2,and observe the biological characteristics.Methods The percentage of CD44＋EpCAM＋ cancer stem cells which were isolated by multi-combination method was evaluated by flow-cytometry.The biological characteristics of the stem cells were analyzed by cell cycle analysis,in vitro invasion assay and in vivo tumorigenicity assay.Results The percentage of CD44＋EpCAM＋ cancer stem cells was 89.57%,and their capabilities of generation,invasion,tumorigenicity were higher than non-cancer stem cells（P 0.05）.Conclusion CD44＋EpCAM＋ cancer stem cells showed low generation,high invasiveness,high tumorigenesis,and they could be isolated by multi-combination.

Objective To investigate the clinical effect of QianlieShutong capsule in combination with tamsulosin in the treatment of benign prostatic hyperplasia ( BPH) and its effects on serum testosterone and estrogen.Methods 114 cases of BPH were randomly divided into control group (n=54) and observation group (n=60).The control group were treated with tamsulosin, while the observation group were treated with QianlieShutong capsule in combination with tamsulosin.Then, the curative efficacy, improvement of related indexes, level of serum testosterone, estrogen and adverse reactions were compared.Results The observation group had a total therapeutic efficacy ratio of 95.0%, which was statistically higher than that of 83.3% in control group (P<0.05).In comparison with control group after the treatment, international prostate symptom score (IPSS), residual urine (Ru), prostate volume (PV) in observation group statistically decreased, and maximum flow rate (Qmax), level of estrogen increased (all P<0.05), while level of testosterone in two groups was statistically same.During the treatment, there’ s no case of severe adverse reaction.And the incidence of adverse reactions in the observation group was 13.3%, which was statistically same with that of 11.1% in control group.Conclusion Combined treatment of QianlieShutong capsule and tamsulosin is effective for BPH, which can significantly increase curative efficacy, improve related indexes and regulate level of sexual hormone with minor adverse reactions.

Objective To investigate the effect of daurinoline on basilar artery vascular smooth muscle. Methods The tension of isolated basilar artery ring of rabbit was measured. The effects of daurinoline on the basilar artery contracted by methoxamine,5-hydroxytryptamine(5-HT),KCl and Histamine( His)were also examined. Dose-response curves of 5-HT and KCl were observed as well. Results Daurinoline exerted obvious relaxation effect on the basilar artery vascular ring contracted by methoxamine,5-HT,KCl and His in a concentration-dependent manner. IC50 of daurinoline in methoxamine,5-HT,KCl and His-treated rabbits was 8.67×10-5,1.78×10-6,6.79×10-7 and 4.98×10-4 mol·L-1,respectively. The change of concentration-response curves of methoxamine,5-HT,KCl and His showed that daurinoline was a non-competitive antagonist. Conclusion Daurinoline exerts marked relaxation effect on basilar artery of rabbits through non-competitive antagonism. The mechanism of relaxation action may be related to blockage of voltage-dependent or receptor-dependent calcium channels.

AIM:To observe the changes of nuclear factor-?B (NF-?B) activity and inducible nitric oxide synthase (iNOS) expression in hypoxic pulmonary hypertension(HPH). METHODS:The rat model of HPH was used. The NF-?B activity and iNOS expression in lung tissue were determined by immunohistochemistry(IHC), in situ hybridization (ISH), RT-PCR and Western blot.RESULTS:ISH showed that iNOS mRNA expression in intraacinar pulmonary arteriole (IAPA) in H28 d group(hypoxic treatment for 28 days) was stronger than that in normal group, H5d group and H14 d group. RT-PCR showed that the iNOS mRNA in H28 group was 2.1 times, 1.9 times and 1.8 times higher than that in normal group, H5d group and H14 d group, respectively. The nucleic anti-NF-?B stain was observed in H28 d group, which was significantly stronger, but the I-?B amount was 2.8 times, 2.7 times and 2.5 times lower than that in normal group, H5d group and H14 d group, respectively. CONCLUSION: The activity of NF-?B was correlated with the hypoxic pulmonary vessel structural remodeling and iNOS expression.

Objective To explore the effect of several cytokines, including interferon-γ, interleukin-10 and interlekin-4, on promoter activity of human BAFF (B-cell activating factor belonging to tumor necrosis factor family) gene. Methods A construct of phBAFF 1.02 containing sequence form -1349 bp to -329 bp of human BAFF gene, linking with chloramphenicol acetyltransferase (CAT) as reporter gene, was transiently transfected into human HL-60 cells, a kind of myeloid tumor cell lines. The cells were subsequently treated with IFN-γ, IL-10 and IL-4, and the CAT activity was assessed 24 hours after stimulation with each cytokines. Results IFN-γ of 5 ng/ml, IL-10 of 100 ng/ml could increase the CAT activity of phBAFF 1.02 to 4.18 and 2.13 folds respectively compared to the control. IL-4 at 100 ng/ml had no effect on promoter activity of human BAFF gene. Combination of IFN-γ, IL-10 and IL-4 could increase the CAT activity of phBAFF 1.02 to 3.41 and 1.58 folds respectively compared with controls. Conclusion IFN-γ and IL-10 can increase the promoter activity of human BAFF gene. IL-4 treatment can not affect the CAT activity driven by BAFF promoter. However, IL-4 can decrease the upregulating effect of IFN-γ and IL-10 on phBAFF1.02. These provide essential evidence for future study on the interaction mechanism of cytokines and BAFF in autoimmune diseases.

Objective To explore the effect of IFN-γ, IL-10 and IL-4 on B cell activating factor (BAFF) expression in human HL-60 cells, a kind of myeloid tumor cell lines, and its possible regulation mechanism. Methods Cultured human HL-60 cells were treated with IFN-γ, IL-10 and IL-4 for 1-3 days. The expression of membrane-bound BAFF on HL-60 cells was examined by flow cytometry, the amount of soluble BAFF was detected by ELISA assay, and the level of BAFF mRNA was tested by real-time PCR method. A functional 1021 bp fragment of the 5'-tlanking region of the human BAFF gene (-1349 to -329 bp) was cloned and investigated with serial 5'-deletion. The 5'-deleted promoters were recombinated with chloramphenicol acetyltransferase (CAT) as reporter gene. These five recombinant plasmids were transiently transfected to HL-60 cells with liposomal transfectian method. Promoters activities were determined by CAT reporter gene assay(CAT-ELISA) in those transfected cells treated with different cytokines. Results The results showed that the expression of membrane-bound BAFF, soluble BAFF and BAFF mRNA in human HL-60 cells were significantly elevated (P ＜ 0. 05) after incubated with IFN-γ and IL-10. In addition, IFN-γ and IL-10 showed significantly (P ＜ 0. 05) increased effects on promoter activity in human BAFF gane. And the cytokines-responsive sequences were located between -929 and -719 bp of the BAFF promoter region. Conclusion The enhancement of IFN-γ and IL-10 on BAFF expression and synthesis were regnla-ted by promoter activation. Our in vitro studies also raise the possibility to investigate the mechanisms regula-ting BAFF expression in other tumor cells of myeloid origin under pathological circumstances.

OBJECTIVE To investigate the antimicrobial-resistant genes and consanguinity in multi-resistant Pseudomonas aeruginosa(MRPA) isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Thirty strains of MRPA were isolated from hospitalized patients between Sep 2003 and Oct 2004.Twenty four kinds of genes of blaTEM,blaSHV,blaOXA-10 group,blaPER,blaVEB,blaIMP,blaVIM,blaGES,blaCARB,blaDHA,blaMIR,aac(3)-Ⅰ,aac(3)-Ⅱ,aac(3)-Ⅲ,aac(3)-Ⅳ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ,aph(3′)-Ⅵ,oprD,qacE△1-sul1,catB,and cml1 were analyzed by PCR and verified by DNA sequencing.Resistant-genes cluster analysis was performed by Average.RESULTS In 30 strains of MRPA the positive rate of genes of blaTEM,blaOXA-10 group,blaCARB,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ,qacE△1-sul1,and cml1 were 66.7%,3.3%,3.3%,76.7%,3.3%,33.3%,53.3%,26.7%,83.3%,and 3.3%,respectively,and the deficiency rate of oprD gene was 90.0%.The gene of blaOXA-10 group was sequenced and determined to be blaOXA-10 subtype ESBL gene.But the rest of genes were all negative.According to the cluster analysis of resistant-gene,30 strains of MRPA isolated could be classified into four subgroups,which were caused by the infection in hospital.CONCLUSIONS At least 10 kinds of antimicrobial-resistant genes exist in MRPA isolates,and the deficiency rate of oprD gene is very high.MRPA can induce clone transmitted hospital infection and it has fulminant prevalence.