Obejective To investigate the effect of exogenous microRNA 136(mir136) on contents of S100 calcium binding protein A6 (S100A6) and proliferation in human colon cancer cell line HCT116 through constructing a eukaryotic expression vector of mir136 (pcDNA-mir136) and cationic liposome-mediated transfection.Methods Human genomic DNA was extracted from HEK293 cells and the DNA fragment containing mir136 precursor sequence was amplified by PCR and cloned into pcDNA to construct pcDNA-mir136. The plasmid was transfected into HCT116 cells via cationic liposome. The content of mir136 was determined by Real-time PCR, S100A6 protein was examined by Western blotting analysis, and apoptosis was detected by Flow Cytometry at 48 h after transfection. The proliferation ability of the tumor cells was examined by CCK-8 at 24 and 48 hours after transfection. Results The eukaryotic expression vector of mir136 was successfully constructed and transfected into HCT116 cells. Expression of mir136 was significantly higher in the transfected group than that in the untransfected group (P<0.01) and the content of S100A6 protein was significantly lower than that of the untransfected group (P<0.05) 48 h after transfection. Meanwhile, cell proliferation activity of the transfected group was significantly lower than that of the untransfected group (P<0.05) and cell apoptosis was significantly increased compared with the untransfected group (P<0.01).Conclusion Mir136 may inhibit proliferation and induce apoptosis of HCT116 cells by inhibiting the expression of S100A6 gene.

OBJECTIVE To study the isolation and resistance tendency of Acinetobacter calcoaceticus-baumannii to antimicrobial agents from 1998 to 2004 to provide valuable data for infection prevention and therapy. METHODS We reviewed the isolation rates,distribution in clinical specimens and wards,and the resistance rates of(A.calcoaceticus-baumannii)to 14 kinds of antimicrobial agents from 1998 to 2004. RESULTS There was an increasing tendency of isolation rates of A.calcoaceticus-baumannii every year,which was 0.18% in 1998 but 1.48% in 2004.In the seven years,there was the highest isolation rate of 70.58% in specimens from respiratory tract,the next was from the urine(9.42%),and blood(4.63%).Concerning the wards distribution,ICU had the highest rate of 47.28%.In 1998,A.calcoaceticus-baumannii had resistance rates more than 50% only to one kind of antimicrobial agents(aztreonam),but in 2004,it had increased to thirteen kinds(except cefoperazone/sulbactam).About the fourteen kinds of antimicrobial agents we inspected,that were increased in their resistance rate.The highest increasing of resistance rate was ceftazidime from 11.1% in 1998 to 88.9% in 2004,the imipenem was second for 0.0% to 64.8%,and the third was sulfamethoxazole/trimethoprim form 0.0% to 64.0%,while there still was an increasing resistance tendency to them. CONCLUSIONS The clinical isolation rate of A.calcoaceticus-baumannii is increasing,and it has higher resistance rates to many antimicrobial agents as well as an increasing resistance tendency to relatively susceptive antimicrobial agents every year.So physicians should prescribe on the basis of antimicrobial agents susceptibility tests in vitro.

OBJECTIVETo summarize experiences of aortoiliac reconstruction through retroperitoneal approach.

METHODSTwenty-eight patients underwent retroperitoneal aortoiliac reconstructions, including aortic aneurysmectomy with graft replacement, aortic endarterectomy with patch angioplasty, thoraco-abdominal aortic bypass, resection of retroperitoneal mass with ilio-femoral bypass, iliac aneurysmectomy with aorto-external iliac artery bypass, removal of aortoiliac foreign body, common iliac endarterectomy, aorto (ilio)-femeral bypass and common ilio-femo-popliteal bypass. Drainage tubes were placed retroperitoneally in 24 cases.

RESULTSAll operations in this group were successful without perioperative death. The volume of intra-operative bleeding was 100-400 ml (mean 240 ml). Blood transfusion were employed in 2 cases. Retroperitoneal drainage was 50-170 ml (mean 85 ml). Naso-gastric tubes were removed 28 h on average after operation. All patients recovered uneventfully except that cardiac insufficiency, stress ulcer and retroperitoneal hematoma were present in 3 patients respectively. Twenty-two patients were followed up from 3 months to 2.5 years. One patient died of AMI 2 years after operation. One patient receiving ilio-femo-popliteal bypass was found to have occlusion of femo-popliteal segment of prosthetic graft. One patient developed brain hemorrhage 1.5 years postoperatively. All the other followed-up patients were living well.

CONCLUSIONRetroperitoneal approach, not violating the peritoneal cavity, offers certain physiological advantages associated with minimal disturbance of gastrointestinal and respiratory function, thereby decreasing respiratory complications and postoperative ileus, avoiding intra-abdominal adhesions with their attendant risk of early and late small bowel obstruction. It proved to be a simple and safe alternative for surgical treatment of aortoiliac diseases.

Adult ; Aged ; Aorta, Abdominal ; surgery ; Aortic Aneurysm, Abdominal ; surgery ; Arterial Occlusive Diseases ; surgery ; Endarterectomy ; Female ; Femoral Artery ; surgery ; Humans ; Iliac Aneurysm ; surgery ; Iliac Artery ; surgery ; Male ; Middle Aged ; Retroperitoneal Space ; surgery ; Retrospective Studies ; Treatment Outcome ; Vascular Surgical Procedures ; methods

BACKGROUNDTo investigate the immunological and virological efficacy of the therapeutic vaccine HBV CS1, a recombinant fusion protein which is composed of HBV core aa 1-155 plus PreS1 aa 3-55,against chronic HBV infection.

METHODSHBV transgenic mice were immunized with HBV CS1(5 ug) emulsified in equal volume of complete Freund adjuvant on day 0, followed by a second vaccination with HBV CS1(5 ug) emulsified with incomplete Freund adjuvant on days 21. Mice of control group were mock-vaccinated with PBS plus complete Freund adjuvant/incomplete Freund adjuvant. The splenocytes of individual mouse were subjected to T cell proliferation assays by using 3Hg thymidine, HBsAg and HBV DNA in sera of mice were detected by ELISA and quantitative PCR, respectively.

RESULTSHBV CS1 specific T cell response were induced in mice immunized with HBV CS1, with the titer of HBsAg and the level of HBV DNA decreased significantly after twice immunization with HBV CS1, while the control group almost remained the same.

CONCLUSIONHBV CS1 has the immunological and virological efficacy against chronic HBV infection in HBV transgenic mice; HBV CS1 could represent candidate vaccine for further studies on its role as therapeutic vaccine against HBV chronic infection in human.

Animals ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B Core Antigens ; genetics ; immunology ; therapeutic use ; Hepatitis B Surface Antigens ; genetics ; immunology ; therapeutic use ; Hepatitis B Vaccines ; genetics ; immunology ; therapeutic use ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Immunization ; Male ; Mice ; Mice, Transgenic ; Protein Precursors ; genetics ; immunology ; therapeutic use ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; therapeutic use

OBJECTIVETo observe the efficacy and safety of vardenafil in the treatment of erectile dysfunction (ED) in aged men with diabetes mellitus (DM).

METHODSOne hundred outpatients with diagnosed ED (40 diabetic and 60 non-diabetic) received vardenafil at the initial dose of 20 mg and sustained dose of 10 mg once a week for 8 weeks, and their erectile functions were evaluated by IIEF and EQS.

RESULTSThe scores on IIEF and EQS in the diabetic ED group were 18.9 +/- 0.2 and 25.1 +/- 1.4 after the vardenafil treatment, significantly higher than 8.1 +/- 0.5 and 9.1 +/- 1.3 before the treatment (P < 0.01), and the non-diabetic group scored 21.1 +/- 0.2 and 34.2 +/- 1.2 on IIEF and EQS after the treatment, as compared with the statistically lower scores of 10.1 +/- 0.3 and 10.1 +/- 1.7 before it (P < 0.01). The total rate of effectiveness was 65% in the diabetic and 73.30% in the non-diabetic group, with statistical differences (P < 0. 05).

CONCLUSIONVardenafil can significantly improve erectile function and is well tolerated in the aged males with diabetic ED.

Aged ; Aging ; physiology ; Diabetes Mellitus, Type 2 ; complications ; Erectile Dysfunction ; drug therapy ; etiology ; physiopathology ; Humans ; Imidazoles ; therapeutic use ; Male ; Middle Aged ; Phosphodiesterase Inhibitors ; therapeutic use ; Piperazines ; therapeutic use ; Sulfones ; therapeutic use ; Treatment Outcome ; Triazines ; therapeutic use ; Vardenafil Dihydrochloride

Objective To investigate the frequency of CD4~+CD25~+FoxP3~+regulatory T cells (Treg)and the expression of the functional protein,FoxP3,in patients with active tuberculosis and the relationship between Treg and the pathogenesis of tuberculosis.Methods Forty-five patients with active tuberculosis(including 25 cases of pulmonary tuberculosis and 20 tuberculous lymphadenitis), 20 healthy controls,20 recovered tuberculosis patients and 6 patients with reactive hyperplasia in cer- vical lymph node were enrolled.The frequency of CD4~+ CD25~+ FoxP3~+ Treg in the peripheral blood was measured by flow cytometry.FoxP3 mRNA expression was determined by real-time reverse transcriptase-polymerase chain reaction(RT-PCR)and the expression of FoxP3 protein in lymphoid tissues was measured by immunohistochemistry.Results The frequency of natural Treg in the peripheral blood from the patients with active tuberculosis was 2.91%?0.23%,which was signifi- cantly higher than that of healthy control group(1.22%?0.18%)and recovered tuberculosis patients(1.50%?0.17%,P

Objective To evaluate the condition of rebuilding stove for preventing coal-buming fluorosis in Ankang City of Shaanxi Province in 2006.Methods Guided by local health administration department in Shaanxi.five counties-Zhengping,Langao,Hanyin,Ziyang,Shiquan-were chosen as rebuilding stove regions in Ankang City.Fifteen housewives were chosen randomly in each village in each county,they were quizzed for the knowledge of health.Fifty students were chosen randomly in each school in each county,they were quizzed for the knowledge of health too.Data of rebuilding stove were refferred.Results Ninty-seven point two per cent(243/250)of the ovens were appropriately sealed,96.8%(242/250)were efficiently exhausted,the rate of oven correctly used was 97.2%(243/250),the awareness rates of healthful knowledge were 85.6%(214/250)in adults and 97.8%(485/496)in children.Data of rebuilding stove were documented systemically,completely and precisely.ConelusionFive model counties in Ankang City have fulfilled the demand.

Objective To mvesugate the Tate of YMDD mutation accompamed with pre-core(region and core promotor region mutation and the clinical significance.Methods YMDD mutation and pre-core(at 1896 nu- cleotide)region and core promotor region(at 1762.1764 nucleotide)mutation were detected from the 122 patients with chronic hepatitis B virus after receiving lamivudine treatment above 6 months.Results 40 cases were tested for YMDI)mutations in 122 HBV patients with lamivudine treatment,and the positive rate of YMDD mutation was 32.8 %.After YMDD mutation,ALT,AST and HBV DNA of the patients significantly increased(P0.05).Conclusion The patients with YMDD mutation had higher rate of pre-core region(at 1896 nucleotide)and basal core promotor region(at 1762, 1764 nucleotide)mutation than those without YMDD mutation,but there was no correlation between the mutation and the deterioration of disease condition and the bad prognosis.

OBJECTIVETo study the prevalence of hepatitis C virus (HCV) infection and characteristics on molecular biology related to HCV among patients who were enrolled in a Methadone maintenance clinic in Wuhan.

METHODSSerum samples from 332 injection drug users (IDUs) were obtained and anti-HCV IgG was detected by enzyme linked immunosorbrent assay(ELISA), together with 86 anti-HCV positive specimens genotyped. A reverse transcriptase-polymerase chain reaction (RT-nPCR) assay using conserved primers deduced from the core-envelopel (C-E1) region of the HCV genome was employed to amplify a 474 bp fragment. Phylogenetic analysis of the C-E1 sequences was conducted by direct sequencing of the RT-nPCR products and alignment with determined by nucleotide sequencing followed by composition of a phylogenetic tree.

RESULTSThere were 313 cases (94.3%) appeared positive anti-HCV IgG in the 332 patients from a Methadone maintenance clinic in Wuhan. It was demonstrated that there were four different subtypes of HCV in that clinic in Wuhan, including 6a--71 cases (82.5%), 3b--7 cases (8.2%), 1a--5 cases (5.8%) and 1b--3 cases (3.5%).

CONCLUSIONInfection of 6a genotype HCV was predominant in patients from the Methadone maintenance clinic in Wuhan, followed by HCV 3b, 1a and 1b.

Adult ; Antibodies, Viral ; analysis ; China ; Enzyme-Linked Immunosorbent Assay ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; Humans ; Male ; Methadone ; therapeutic use ; Middle Aged ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Substance Abuse Treatment Centers ; Substance-Related Disorders ; drug therapy ; rehabilitation ; Young Adult

Objective To study the pharmacokinetic features of reactive sulfide in rats after oral administration of Cinnabaris. Methods An HPLC coupled with precolumn derivatization method was developed for the pharmacokinetic features study on reactive sulfide in rats after oral administration of Cinnabaris. Results Good linearity (r>0.99) was found for reactive sulfide in plasma in the concentration range of 0.25–15 μmol/L (r>0.99). The LOQ and LOD of the method were 0.1 μmol/L and 0.02 μmol/L, respectively. The intra- and inter-day precision was less than 4.4% and 3.5% respectively, and the accuracy was -9.9%–6.0%. The average recovery rate was 74.9%. 0.6 g/kg Cinnabaris was given the rats for gavage, and the time-course pharmacokinetics parameters were as follows:Cmax(1.33±0.13) μmol/L, tmax(150±34) min, t1/2(323±62) min, AUC0-∞ (5743±297) ng/mL?h. Conclusion A sensitive, robust and accurate precolumn derivatization-HPLC method for the determination of plasma reactive sulfide is developed and validated. The method is successfully applied in the pharmacokinetic features study on reactive sulfide in plasma of rats after administration of Cinnabaris.