Objective To investigate the distribution of hepatitis B virus(HBV) genotypes ,drug resistance situation of hepatitis B patients and the relation between HBV genotype with drug resistance and mutation sites .Methods Three hundred serum samples of HBV patients were collected and HBV‐DNA was extracted by adopting the centrifugal column method .The HBV genotype and drug resistant mutant were detected by using PCR‐reverse dot blot hybridization method .Results In 300 cases of HBV‐DNA posi‐tive ,genotye B ,C ,B/C and other undetected genotypes were detected out ,but genotype D was not detected out ,in which genotype C was predominant ,accounting for 81 .8% ;in the HBV patients ,the resistant drugs were dominated by‐lamivudine and telbivudine , accounting for 43 .6% ;the HBV drug resistant mutation genotypes were mainly rt204I(24 .35% ) ,rt204V (17 .39% ) and rt180M (17 .39% );the drug resistance mutation rates of genotype B and C were 30 .77% and 42 .42% respectively ;the difference was sta‐tistically significant(P<0 .05) .Conclusion HBV genotype C in Dongying area is more than genotype B ,genotype C is prone to produce drug resistance ,rt204I ,rt204V and rt180M gene mutations are common ,lamivudine and telbivudine combined resistance is common ,the suitable treatment scheme should be selected according to genotyping and drug resistance mutations results .

Neonatal nerves exist in tumor tissues,and tumor cells can invade nerve by neurogenesis.Current research shows that cancer cells can secrete cytokines to promote the differentiation and growth of nerve cells,in turn,the differentiation and growth of nerve cells can stimulate tumor development and metastasis.Neurogenesis is one way of interaction bettween tumor cells and microenvironment similar to angiogenesis and lymphangiogcnesis,which has a major impact on biological behavior of tumor cells and prognosis.

OBJECTIVE To evaluate the effect of azithromycin on mycoplasmal pneumonia(MP). METHODS We divided 204 MP cases into treatment and control groups.In treatment group 10mg/(kg?d) ivgtt for 3-5 d,while in control group 30mg/(kg?d) erythromycin dripping iv.qd for 7-10 d and then observe the effect. RESULTS In treatment and controll groups the effective rate was 91.5% and 78%,respectively.The average days of curing for cough were 6.8 days and 8.7 days(P

Objective:To observe therapeutic effect of ambrisentan on Eisenmenger syndrome (ES) .Methods:A total of 79 ES patients ,who hospitalized in Wuhan Asia Heart Hospital from Apr 2011 to Mar 2014 ,were divided into ambrisentan group (n=38 ,received digoxin combined ambrisentan treatment) and routine treatment group (n=41 , received digoxin treatment) ,and both groups were treated for 12 weeks .The 6min walking distance (6MWD) , WHO functional class (WHO FC) of pulmonary hypertension etc .were observed in both groups before and 12-week after treatment .Changes of hemodynamic parameters ,including mean pulmonary artery pressure (mPAP) ,total pulmonary resistance (TPR) and ratio of pulmonary circulating flow to system circulating flow (Qp/Qs) etc .,were observed in ambrisentan group before and after treatment . Results:Compared with before treatment , after 12 weeks ,6MWD significantly rose ,TPR significantly reduced ,Qp/Qs significantly increased and WHO FC signifi-cantly improved in ambrisentan group ,P<0.05 or <0.01. Compared with routine treatment group after 12 weeks , there was significant rise in 6MWD [ (356.8 ± 74.1) m vs .(393.7 ± 69.2) m] ,significant reduction in BNP level [(832.5 ± 422.2) ng/L vs .(646.6 ± 297.1) ng/L] and significant improvement in WHO FC (class I/II ,67.5% vs . 86.8% ) in ambrisentan group ,P<0.05 or <0.01. No patient died ,the adverse reactions were mild in ambrisentan group ;one case died in routine treatment group .Conclusion:Oral ambrisentan can significantly improve heart func-tion and exercise capacity in patients with Eisenmenger syndrome ,and it possesses good safety and tolerance .

AIM:To investigate the effect of 18 alpha-glycyrrhetinic acid (18α-GA) on delaying the senescent progress and promoting the proliferation in late-passage bone marrow mesenchymal stem cells ( BMSCs ) .METHODS:Late-passage BMSCs were incubated with 2.0 mg /L 18α-GA or the same volume of DMSO for 30 d, and the cells were harvested to determine the proteasome activity .The expression of senescence-related proteins p53, p21 and p16 was detec-ted by senescence-associated β-galactosidase ( SA-β-Gal) staining and Western blot .The cell proliferation , the expression level of cell cycle-related proteins and cell cycle distribution of the cells were measured by CCK -8 assay, BrdU incorpora-tion, Western blot and flow cytometry.RESULTS:Compared with DMSO group, the proteasome activity in 18α-GA group increased significantly by about 0.2 times (P<0.01).SA-β-Gal-positive cells in 18α-GA group decreased, and cell stai-ning was lighter.The contents of p53 and p21 in 18α-GA group were decreased (P<0.05).The results of CCK-8 assay showed that the A value in 18α-GA group was 0.3 times higher than that in DMSO group (P<0.01).BrdU incorporation showed the increased proliferation in 18α-GA group compared with DMSO group ( P<0.05).The cells in G1 phase in 18α-GA group decreased significantly compared with DMSO group , while the cells in S phase increased significantly ( P<0.05).The expression level of cyclin D1 in 18α-GA group was 2.8 times higher than that in DMSO group (P<0.01), and the CDK4 level was 1.4 times higher than that in DMSO group (P<0.05).CONCLUSION:Activation of the pro-teasome activity by 18α-GA delays the aging process in the BMSCs and promotes the cell proliferation via up -regulation of the cell cycle-related proteins .

Objective This study evaluated the effects of combined procedure of surgical treatment and cell transplantation on dilated cardiomyopathy. Methods Eight patients (5 men and 3 women) with moderate to severe mitral regurgitation from end-stage dilated cardiomyopathy underwent surgery. Four patients were in functional class (FC) Ⅳ, six were in FC Ⅲ. There age ranged from 15 to 56 years. The preoperative ejection fraction (EF) ranged from 0.15 to 0.32 (mean 0.26 ± 0.08). Mitral valve replacement was performed in 5 patients and mitral valve repair in 3. Auto-bone marrow monenuclear cells were harvested, isolated, washed, and resuspended for direct injection after surgical procedure. Results All patients survived and were discharged from the hospital. After a mean follow-up period of 18 months ( 12 - 42 months). Echocardiography showed postoperative ejection fraction and wall movement velocity increased after 6 months. Radionuclide ventriculography showed myocardial perfusion improved significantly. Conclusion Combined procedure of surgical treatment and cell transplantation led to significant improvement in cardiac function in patients with dilated cardiomyopathy.

OBJECTIVE:To establish a method for rapid identification and efficient preparation of main ingredients in effective parts of Xinjiang Artemisia rupestris,and provide reference for researching the ethnic medicines. METHODS:LC-HRMS/MS was conducted for the preliminary study of main ingredients in effective parts of A. rupestris. HPLC,UV and MS were used to compare and analyze parts of the compounds and its reference substances,their names were determined. Column separation and preparative liquid chromatography were used for the undetermined compounds to receive monomer rapidly,and the structures were identified. RESULTS:5 compounds were separated from the effective parts,2 of which were identified as artemetin and casticin. A monomer-ic compound was obtained (yield was 0.35 mg/g,the purity was 98.5%),which was confirmed to be 6-demethoxy-4′-O-me-thoxy-capillarisin-7-O-β-D-glucoside by the structure. CONCLUSIONS:The method has achieved rapid separation,identification and preparation of target ingredients,which can be used for the fundamental research of ethnic medicine complex materials.

Objective To synthesize glycylproline p-nitroanilide tosylate as a substrate for glycylprolyl dipeptidyl-aminopeptidase(GPDA),and to study its clinical application.Methods The title compound was prepared by DCC-HOBt synthesis.GPDA in human serum was detected by continuous monitoring method using substrate. Results The substrate with purity of 98.5% was obtained.The linearity of the method was up to 358.1 U/L.Intraassay CV and interassay CV of GPDA in diverse serum samples were 3.01% and 5.04%,respectively.It was shown that GPDA level in patients with hepatic cancer was significantly increased,while those in patients with gastric carcinoma and chronic gastritis were significantly decreased compared with that in the control group in clinical detecting(P

Objective To investigate the distribution and drug resistance of the gram positive bacteria isolates from hospital in Chongqing during 2006-2011 .Methods Non-repetitive gram positive bacterias were collected retrospectively from ten hospitals in Chongqing .Antimicrobial susceptibility testing of all clinical isolates were carried out using Kirby-Bauer(KB) .Results 14 473 strains were isolated .There were 4 840 strains of Staphylococcus aureus (33 .4% ) ,2 566 strains of Staphylococcus epidermidis (17 .7% ) ,1 822 strains of Staphylococcus haemolyticus (12 .6% ) ,1 325 strains of Enterococcus (E .faecalis/E .faecium )(9 .2% ) . Staphylococcus was highly sensitive to nitrofurantoin ,rifampicin ,piperacillin/tazobactam ,linezolid ,teicoplanin and vancomycin ,but their resistance rate to ampicillin/sulbactam ,oxacillin ,penicillin and clarithromycin was high .The resistance rate of E .faecium to the majority of antimicrobial agents was obviously higher than that of E .faecalis ,excluding imipenem ,chloramphenicol and tetracy-cline .Vancomycin and linezolid still remain the strongest antibacterial activity to Enterococcus .Conclusion The distribution and drug resistance of pathogenic bacteria in Chongqing present regional particularity ,and antibiotic resistance is severe .It is significant to strengthen the epidemiology study of pathogenic bacteria and to develop policies to intervene in the use of antimicrobial agents .

Objective To study the effect of astrocyte on synapse formation and the molecular mechanism. Methods Cortical astrocytes were isolated and purified from neonatal rats.On the 2h,7th day,14th day and 21th day after passage,we counted the number of astrocytes and the culture medium(astrocyte-conditioned medium,ACM)was harvested to measure the concentration of estrogen(E 2)by using ELISA techniques.Based on the model of pure culures of neonatal cortical neurons,the experimental groups were designed as follows:1.pure neuron cultures(group N);2.ACM cultures(group A);3.mixed cultures(group M);4.E 2 cultures(group E 2);5.ACM+Tamoxifen(estrogen receptor antagonist)cultures(group A+T);6.Tamoxifen cultures(group T).Then synaptic puncta in every group was stained and counted through immunofluorescence,and we also compared the differences in puncta number among those six groups(at 9th day in culture,number/per neuron). Results The numbers of astrocytes were:1?10 4/ml, 1.1?10 6/ml, 1.4?10 6/ml, 1.5?10 6/ml; The concentrations of E 2 were:(ng/L):0, 117?22, 266?22,252?27 respectively.No estrogen was detected in the primary culture medium.The concenteration of estrogen increased in correspondence with the culturing days and reached the peak around at the 14th day, then decreased gradually but kept at a certain high level,and the numbers of synaptic puncta of per neuron in group N,A,M,E 2,A+T,T were:14?3;79?5;83?8;80?6;32?3;29?3 respectively.The treatment of pure neuron culture with ACM increased the number of synapses on per neuron by up to 6 fold by comparison with pure neuronal culture.Exogenic estradiol added into pure neurons can mimick the effect of the ACM.Tamoxifen which is antagonist of estrogen receptor could decrease the effect of ACM by 75%.Conclusion The astrocytes of neonatal rat cortex do secrete E 2.Astrocyte-derived estrogen may be the molecule regulating the synaptic formation through estrogen receptors.