Objective To investigate the influence of endoscopic retrograde cholangiopancreatography (ERCP) on life quality of patients with common bile duct stones. Methods The life quality of 35 patients, who underwent ERCP because of common bile duct stones, were assessed by GIQLI questionnaires before, 2 and 6 weeks after the procedure. Results The mean GIQLI score of patients before ERCP was 99. 9, which was significantly increased to 112. 2 and 121.9 at 2 and 6 weeks after ERCP (P ＜0. 01 ). At 6weeks after ERCP, the GIQLI score was similar to that of normal population. Conclusion ERCP can improve the life quality of patients with common bile duct stones.

Objective To explore the effects of PP242 on the expression of apoptosis protein Bcl?2 in human lens epithelial cells(HLECs). Meth?ods Immortal HLECs SRA01/04 were cultured and treated with different concentrations of PP242. The cell growth was examined by MTT assay at 24 h and 48 h after PP242 treatment. Real?time quantitative polymerase chain reaction(RT?PCR)and Western blot were adopted to detect the mRNA and protein expression of Bcl?2 respectively. Results The cultured HLECs SRA01/04 were collected and treated with different concentra?tions(100,250,500,750,1 000,1 500 nmol/L)of PP242. After treatment for 24 h and 48 h,the inhibitory rate in each well was determined by MTT assay. The inhibition rate was as below,24 h:7.55%,9.43%,16.98%,22.64%,26.42%,30.19%;48 h:11.11%,23.81%,36.51%,42.86%, 49.21%,63.49%. Compared with the control group,the difference was statistically significant(P<0.05). Compared with the control group(0 nmol/L),the expression of Bcl?2 protein was significantly decreased with the increasing concentrations of PP242(P<0.05),while the expression of Bcl?2 mRNA was inhibited by PP242 based on the results of RT?PCR. Compared with the control group(0 nmol/L),the RT?PCR results of Bcl?2 mRNA were 0.723±0.039,0.517±0.028,0.353±0.052,0.167±0.046,respectively(P<0.05). Conclusion PP242 could inhibit cell proliferation of HLECs in vitro with a concentration and time depended manner. Bcl?2 protein is expressed in HLECs,which could be down regulated by PP242 treatment.

ObjectiveTo construct human embryonic kidney cells (HEK293) modified with human preproenkephalin (hPPE) gene.MethodshPPE gene fragments were obtained from recombinant plasmid pcDNA3.1( + )/hPPE by using restriction endonuelease Hind Ⅲ and Not Ⅰ.Homologous recombination of lentivirus and hPPE gene was produced by using recombinant DNA technology.HEK293 cells were then transfected with the recombinant lentivirus vectors.The expression of hPPE gene in HEK293 cells was detected by Western blot.ResultsThe results of DNA sequencing indicated that the positive clone of recombinant lentivirus was completely consistent with sequencing of hPPE in Genebank.The titer of the concentrated virus was 2.07 × 108 TU/ml.GFP fluorescence was not seen in HEK293 cells transfected with the lentiviral vector under fluorescence microscope.A strong fluorescence was seen in HEK293 cells transfected with Ubc-GFP-L.V.empty viral vector.Positive expression of hPPE was demonstrated in HEK293 cells transfected with lentiviral vector by Western blot.Conclusion HEK293 cells modified with hPPE gene were successfully constructed and the target gene hPPE was stably expressed in HEK293 cells.

Objective To explore the value of determination of combined tumor markers based on artificial neural network (ANN) discrimination model in facilitating the diagnosis of hepatic carcinoma. Methods Serum samples were collected from three groups of subjects, including 50 cases of liver cancer, 40 cases of benign liver disease, and 50 normal controls. The levels of serum alpha fetoprotein (AFP), carbohydrate antigen 125 (CA125) and carcino-embryonic antigen (CEA) were determined by chemiluminescence immunoassay. The level of serum sialic acid (SA) was determined by spectrophotometry, the content of calcium in serum was measured by calcium assay kit (Azo-end method of arsenic HI). Based on the five tumor markers mentioned above as discrimination variables, Fisher discrimination and ANN were applied to set up the intelligent auxiliary diagnostic model. Results By applying the Fisher discrimination model established in present work, the diagnostic sensitivity of liver cancer was 46.1%, the specificity was 98.9%, the accurate rate was 79.3%, the positive predictive value was 95.8%, and the negative predictive value was 76.7% for the three groups. With the application of ANN discrimination model, the diagnostic sensitivity of liver cancer was raised to 96.0%, the specificity 98.9%, the accuracy 94.3%, the positive predictive value 98.0%, and the negative predictive value was 97.8%. Conclusion The diagnostic model based on ANN combined with 5 tumor markers is superior in diagnostic acuity to traditional Fisher discrimination analysis, thus more suitable for clinical data analysis.

Objective To assess the safety, feasibility, short-and mid-term efficacy of wingspan stent for treating patients with symptomatic intracranial artery stenosis. Methods A total of 113 patients with severe symptomatic intracranial stenosis were enrolled and Gateway-wingspan stenting were performed on all patients. The technical success, the pre- and post-stenting stenosis, perioperative complications, clinical outcome and restenosis rates were recorded, and chi-square test was used for analysis of complication rate by comparing our results with the results of Warfarin-Aspirin Symptomatic Intracranial Disease (WASID) study and NIH multi-center Wingspan stenting trial. Results The technical success rate was 99. 1% ( 112/113). The mean pre and post-stent stenoses were (80.7 ± 9.3)% and (27.7 ± 9.7)% (χ2 =9.397,P < 0. 05 ). The total complication rate was 4.4% (5/113 ) during the follow-up ( mean 14. 5 months, range 1-28 months), and the frequency of restenosis was 12. 5% (5/40) at 6 months. The primary endpoint events, ischemic stroke, and lesion-related ischemic stroke were lower in our study (4.5%, 3.5%,3.5% ) compared with the results of WASID trial (21.1%, 20. 4%, 15.0% ,P<0. 05). For those with poor outcome in the three high-risk sub-groups which were with more than 70% stenosis, or last event from the treatment was less than 17 days, or NIHSS was above 1, a better outcome was observed in our group (4. 5% ,4. 7% and 2. 0% in our study, 19.0%, 17.0% and 19. 6% in previous study, P < 0. 05). The medium-term efficacy in this group (4. 5% ) significantly improved compared with NIH study ( 14. 0% ,P <0. 05 ). Conclusions Wingspan stenting for symptomatic intracranial arterial stenosis is with good safety,feasibility and low perioperative stroke rate and mortality. The incidence of primary endpoint events and the ischemic events are lower than those of medication group, and the efficacy of stenting is significantly better than medication even in high-risk population.

Objective To evaluate the mid-and long-term follow-up outcome of revascularization and stenting of nonacute intracranial vertebrobasilar artery occlusion. Methods Consecutive data of 27 patients who suffered from nonacute intracranial vertebrobasilar artery occlusion beyond 24 hours and underwent endovascular revascularization, were retrospectively collected and analyzed. Complications and recurrent ischemic events during the follow-up period were recorded. The modified Rankin scale(mRS) scores were used and compared between pre-and postoperation. Results All 27 patients except one(96.3%) obtained successful recanalization. After the procedure, 13 patients showed improvement, 11 were stable, and 3 worse. The decline of median mRS scores, which was 4(interquarter range-IR, 2-5) preoperatively and 3(IR, 1-5) on discharge. Five patients suffered from procedural complications and three of them resulted in aggravation. Nineteen patients received imaging follow-up during the median 10 months, 6 restenosis occurred and 3 of them were symptomatic. During median 55 months clinical follow-up after operation, 2 ipsilateral stroke and 2 ipsilateral transient ischemic attack occurred. The mRS scores decreased significantly in the first one year after procedure. Conclusions Revascularization and stenting of nonacute intracranial vertebrobasilar artery occlusion can prevent recurrent ipsilateral ischemic event and improve disability recovery in the first one year.

Objective To evaluate the release of leucine-enkephalin (L-EK) in the cerebrospinal fluid induced by intrathecal human embryonic kidney (HEK293) cells modified with human preproenkephalin (hPPE)gene and the analgesic efficacy of L-EK for bone cancer pain.Methods Forty CIBP female Sprague-Dawley rats were randomized into transplantation (CIBP+ hPPE/HEK293,n =20) and control (CIBP + HEK293,n =20)groups using a random number table.At 1 day before inoculation of cancer cells (T1,baseline) and 8,15,21,25,32 and 35 days after inoculation (T2-7),thermal paw withdrawal latency (TWL) was measured,and the number of licking/biting the claw on the transplantated side and degree of hindlimb limping during free activities were recorded.After observation at T4,10 rats were chosen from each group and sacrificed and the cerebrospinal fluid of rats was collected in an ice bath for detection of hPPE expression using radioimmunoassay.Results Compared with control group,TWL was significantly prolonged,the concentration of L-EK in the cerebrospinal fluid was increased,and the number of licking/biting the claw on the transplantated side and degree of hindlimb limping during free activities were decreased at T4-7 in transplantation group.Conclusion Intrathecal HEK293 cells modified with hPPE gene can continuously secrete L-EK and mitigate bone cancer pain.

Objective:To observe influence of statins on antiplatelet activity of clopidogrel and provide basis for ra‐tionality of statins combined clopidogrel treatment .Methods :According to random number table ,a total of 90 pa‐tients diagnosed as acute coronary syndrome were equally divided into clopidogrel group ,clopidogrel + simvastatin group and clopidogrel + pravastatin group . Three groups received corresponding routine medication treatment . Plasma levels of platelet αgranule membrane protein (CD62P) ,lysosomal granule membrane glycoprotein (CD63) and maximum platelet aggregation rate (MPAR) were measured and compared among three groups before and 3d af‐ter treatment .Results:Compared with before treatment ,after treatment ,there were significant reductions in plas‐ma levels of CD62P and CD63 and MPAR in three groups , P<0.01 all .After treatment ,there were no significant difference in plasma levels of CD62P [ (14.63 ± 3.45) ng/ml vs .(14.14 ± 4.32) ng/ml vs .(14.59 ± 4.23) ng/ml] , CD63 [ (26.32 ± 10.43) ng/ml vs .(27.04 ± 10.75) ng/ml vs .(27.29 ± 9.27) ng/ml] and MPAR [ (28.62 ± 17.68)% vs .(28.38 ± 16.43)% vs .(29.13 ± 14.23)% ] among clopidogrel group ,clopidogrel + simvastatin group and clopidogrel + pravastatin group ,P>0.05 all .Conclusion:Short‐term and routine dose of statins combined clo‐pidogrel is feasible in treatment of acute coronary syndrome .The combined use of them will not affect antiplatelet function of clopidogrel .

Objective:To explore protective effect of atorvastatin on blood vessels in early stage of atherosclerosis (AS).Methods:A total of 120 patients without AS plaques,who had >2 cardiovascular risk factors and received control cardiovascular risk factors therapy,were randomly divided into four groups:control group (did not receive atorvastatin),atorvastatin 5mg group,10mg group and 20mg group (received corresponding dose of atorvastatin). All patients were followed up for six months,changes of thromboxane B2 (TXB2),6-Keto-prostaglandin F1α (6-Keto-PGF1α),brachial-ankle pulse wave velocity (baPWV),ankle brachial index (ABI)and intima-media thickness (IMT)were observed.Results:There were no significant changes in ABI and IMT between before and after treat-ment among four groups (P >0.05 all).Compared with baseline,TXB2、baPWV levels significantly rose,6-Keto-PGF1αlevel significantly decreased after treatment in control group and 5mg group;in contrast,TXB2、baPWV lev-els significantly decreased,6-Keto-PGF1αlevel significantly rose after treatment in 10mg group and 20mg group(P <0.05~ < 0.01).After treatment six-month,compared with control group and 5mg group,the TXB2 [(148.3 ± 29.2)pg/ml,(142.3±30.6)pg/ml vs.(111.5±22.8)pg/ml,(104.9 ± 17.4)pg/ml]、baPWV[(1621.1 ± 136.1) cm/s,(1597.7±125.3)cm/s vs.(1232.9±132.3)cm/s,(1178.2±155.1)cm/s]levels significantly decreased,6-Keto-PGF1α[(104.7±66.1)pg/ml,(102.2±70.3)pg/ml vs.(132.8±48.3)pg/ml,(139.1±66.3)pg/ml]level significantly rose(P <0.05~<0.01)in 10 mg group and 20 mg group.Conclusion:Atorvastatin has protective effect on blood vessels in early stage of atherosclerosis,and 10mg atorvastatin may be the minimum effective dosage to protect blood vessels.

Objective To investigate the prevalence of plasmid-mediated quinolone resistance (PMQR) genes among clinical isolates of Klebsiella pneumoniae (K.pneumoniae) in pediatrics.MethodsA total of 131 non-duplicate clinical isolates of K.pneumoniae were collected in the Affiliated Children′s Hospital of Capital Institute of Pediatrics from 2010 to 2012.PMQR genes [qnrA, qnrB, qnrS, aac(6′)-Ⅰb-cr and qepA], mutations in the quinolone resistance-determining region (QRDR) and extended spectrum β-lactamases (ESBLs) genes in those strains were analyzed by PCR.Minimum inhibitory concentrations (MIC) of different antibiotics against those K.pneumoniae strains were determined by broth microdilution method and E-test according to the guidelines issued by the Clinical and Laboratory Standards Institute (CLSI).Transferability of the PMQR genes was examined by conjugation test with the sodiumazide-resistant Escherichia coli J53.Results Among the 131 isolates, 9.92% were resistant to quinolone and 30.5% were positive for PMQR genes, including 6.87% harboring qnrB gene, 22.9% harboring qnrS gene and 4.58% harboring aac(6′)-Ⅰb-cr gene.Neither qnrA-positive nor qepA-positive strain was detected.Among these PMQR genes-positive isolates, 90% were ESBLs-producing strains and two presented mutations in gyrA and parC genes.Conjugation test showed that these PMQR genes could be transferred horizontally and the ciprofloxacin resistance increased 2 to 32 folds in transconjugants.Conclusion This study indicates that the PMQR gene-carrying rate is high in K.pneumoniae strains isolated in paediatrics in China.Most of the PMQR gene-positive strains are also ESBLs-producing strains.The PMQR genes could be transferred horizontally in bacteria.