To construct a cDNA subtractive library of genes transactivated by hepatitis C virus core protein with suppression subtractive hybridization technique. mRNA was isolated from HepG2 cells transfected with pcDNA3 1(-)-core and pcDNA3 1(-) empty vector,respectively, then cDNA was synthesized. After restriction enzyme RsaI digestion, small sized cDNA were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, and then it was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain JM109. The cDNA were sequenced and analyzed in GenBank with Blast search after PCR. The subtractive library of genes transactivated by HCV core protein was constructed successfully. The amplified library contained 233 positive clones. Colony PCR product showed that 213 clones contained 100~ 1 000 bp inserts. Sequence analysis was performed in 63 clones. Six of the sequences were unknown genes before. The full length sequences were obtained with bioinformatics method,which had been accepted by GenBank. It suggested that six novel cDNA sequences might be target genes transactivated by HCV core protein.

Wnt/β-catenin signaling pathway plays an important role in the proliferation, growth, invasion, and metastasis of human cancers. Moreover, β-catenin/T-cell factor 4 (TCF4) interaction regulates the transcription of the key oncogenes in Wnt/β-catenin signaling pathway. Therefore, β-catenin/TCF4 interaction would be a promising therapeutic target for the development of highly selective anticancer agents. At present, most ongoing small-molecule inhibitors targeting β-catenin/TCF4 interaction, including PKF222-815, iCRT3/5/14, LF3, and sanguinarine, have been developed in preclinical studies for human cancer therapeutics. In this review, we summarized the research advances of up-to date inhibitors targeting β-catenin/TCF4 interaction, including the molecular structure and cellular functions of β-catenin in canonical Wnt signaling pathway. This review holds a hopeful avenue for the development of novel and highly selective Wnt inhibitors targeting β-catenin/TCF4 interaction for future anticancer strategy.

ObjectiveTo investigate serum lipopolysaccharide (LPS) level in people with different glucose tolerances and to explore the relationship between LPS and insulin resistance/β-cell secretory function.Methods Sixty-seven subjects were recruited,including 23 with newly diagnosed type 2 diabetes ( T2DM),21 impaired glucose tolerance ( IGT),and 23 normal glucose tolerance (NGT).Serum LPS was assayed by limulus amebocyte lysate test ;expression of LPS toll-like receptor 4 (TLR4) on surface of plasma monocytes was measured by flow cytometric assays,and the changes of LPS levels by 0.5 hours and 2 hours after a high-fat diet were detected.Insulin resistance was evaluated by homeostasis model assessment for insulin resistance (HOMA-IR); β-cell secretory function was evaluated by homeostasis model assessment for β3 cell function ( HOMA-β )/HOMA-IR,increment in insulin in the first 30 minutes/increment in glucose in the first 30 minutes ( AIns30/ΔG30)/HOMA-IR,AUCIns120min/HOMA-IR.Results2 h LPS levels after a high-fat diet were significantly higher than fasting LPS levels [ NGT:0.96(0.33,0.99)vs 0.62 (0.22,0.64),IGT:1.08(0.53,1.22)vs 0.71 (0.39,0.82),T2DM:1.23 (0.62,1.43)vs 0.86( 0.45,0.94 ),EU/ml,all P＜0.01 ].Fasting,0.5 h,and 2 h LPS levels and fasting TLR4 levels of T2 DM group and IGT group were respectively higher than those of NGT group [ fasting LPS:0.86( 0.45,0.94 ),0.71 ( 0.39,0.82 ) vs 0.62(0.22,0.64),EU/ml;0.5 h LPS:1.10(0.55,1.18),0.84(0.50,1.07) vs 0.73(0.31,0.76),EU/ml;2 h LPS:1.23(0.62,1.43),1.08(0.53,1.22)vs 0.96(0.33,0.99),EU/ml; fasting TLR4:36.96( 17.22,55.19),30.34 ( 15.00,45.18 )vs 15.66 (6.09,9.76),MIF/105 cells,all P＜0.01 ].Fasting LPS,AUCLPS 120 min,and fasting TLR4 were positively correlated with insulin resistance index and negatively correlated with β-cell secretory function index ( P＜0.05 ).Multiple linear regression analysis showed that fasting LPS was an independent correlative factor of HOMA-IR and 0.5 h LPS was an independent correlative factor of (AIns30/AG30)/HOMA-IR and AUCIns Ins120min/HOMA-IR.ConclusionPeople with different glucose tolerances show differed LPS levels and its receptor TLR4 levcls,both of which are correlated with insulin resistance and β-cell secretory function,suggesting that LPS is associated with the pathogenesis of abnormal glucose regulation.

Objective To investigate the influence of laparoscopic D3 lymphadenectomy combined with pelvic autonomic nerve preservation on the urinary function of male patients with rectal cancer. Methods From August 2006 to August 2007, 119 male patients with rectal cancer who had been admitted to Southwest Hospital were assigned to open surgery group (n=59) and laparoscopic group (n=60) according to the random number table. Three months after the operation, the urinary function of patients was assessed by urodynamics investigation and international prostate symptom score (IPSS). Differences in measurement data were compared with paired t test. Results There was no significant difference in IPSS between laparoscopic group (10.9±2.9) and open surgery group (11.5±3.1) (t=-1. 309, P>0.05). The maximum flow rate, voided volume, residual urine volume, detrusor contraction pressure and maximum urethral pressure were 15.2 ml/s, 150.1 ml, 6.1 ml, 43.3 cm H2O (1 cm H2O=0.098 kPa) and 77.5 cm H2O in laparoscopic group, and 15.0 ml/s, 140.9 ml, 6.4 ml, 45.6 cm H2O and 72.3 cm H2O in open surgery group, with no statistical difference between the 2 groups (t=1.22, -2.12, -1.73, -1.35, -1.64, P>0.05). Conclusions Laparosceopic D3 lymphadenectomy combined with pelvic autonomic nerve preservation is relatively safe in preserving urinary function, and its efficacy is comparable to that of open surgery.

Adolescent ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; Cyclophosphamide ; administration & dosage ; Diagnosis, Differential ; Doxorubicin ; administration & dosage ; Female ; Hodgkin Disease ; pathology ; Humans ; Lung Neoplasms ; pathology ; Lymphoma, B-Cell ; drug therapy ; pathology ; surgery ; Male ; Mediastinal Neoplasms ; drug therapy ; pathology ; surgery ; Mediastinum ; surgery ; Neoplasm Invasiveness ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; pathology ; Prednisone ; administration & dosage ; Thymoma ; pathology ; Thymus Neoplasms ; pathology ; Vincristine ; administration & dosage

Objective To observe the safety and efficacy of preoperative Ticagrelor loading in emergency percutaneous coronary intervention (PCI)for acute ST-segment elevation myocardial infarction(STEMI).Methods A total of 213 patients with acute STEMI before undergoing emergency PCI were randomly divided into Ticagrelor group(n =105)receiving 180 mg Ticagrelor loading dose,then 90 mg twice daily and Clopidogrel group(n =108) receiving 600 mg of Clopidogrel,then 75 mg once daily.Emergency PCI postoperative coronary artery TIMI flow grade and the change of incidence of no reflow,platelet aggregation rate,incidence of bleeding events and the incidence of major adverse cardiovascular events(MACE) were compared between two groups.Results The rate of no-reflow was 7.6 ％ (8 cases)in Ticagrelor group,and 16.7 ％ (18 cases) in Clopidogrel group(x2 =3.26、P=0.030).Platelet aggregation rates at 1 h and 24 h after treatment were (55.6±4.3)％ and (48.6 ± 4.1) ％ respectively in Ticagrelor group,and (63.6 ± 3.8) ％ and (57.6 ± 3.6) ％ respectively in Clopidogrel group,which showed that platelet aggregation inhibition effect was better in Ticagrelor than in Clopidogrel (t =14.40、17.20,both P =0.001).Two groups had no major life-threatening bleeding events.Bleeding incidence had no statistically significant difference between two groups(x2 =0.14,P =0.710),and the incidence of cardiovascular adverse events showed no statistically significant difference(x2 0.04,P 0.840)between the 2 groups.Conclusions Preoperativeticagrelor loading treatment in emergency PCI therapy for acute ST segment elevation myocardial infarction shows stronger antiplatelet aggregation function,significantly improve postoperative TIMI flow,and does not increase the incidence of bleeding events.

Objective To study the correlation between human leucocyte antigen-G (HLA-G) 14 bp insertion/deletion (I/D) polymorphism and susceptibility to non-small cell lung cancer (NSCLC) as well as poor prognosis in NSCLC.Methods A total of 113 patients with NSCLC and 150 age-and sex-matched healthy subjects were genotyped by PCR to analyze the HLA-G 14 bp insertion/deletion polymorphism in them.Epidermal growth factor receptor (EGFR) gene mutation in patients with NSCLC was detected by using amplification refractory mutation system (AMRS).Expression of HLA-G in NSCLC tissues was detected with immunohistochemistry.All patients with NSCLS were followed up to collect survival data, which were further analyzed with Kaplan-Meier method.Results The frequency of HLA-G 14 bp D/D genotype was significantly higher in the patients with NSCLC than that in the healthy subjects (x2=3.907, P=0.048, OR=1.66).Among the patients with NSCLC, HLA-G 14 bp I/I genotype carriers had a shorter overall survival time as compared with that of HLA-G 14 bp I/D or HLA-G 14 bp D/D genotype carriers (P=0.005).Patients who received chemotherapy or radiation had significantly shorter survival time than those received EGFR-targeted therapy (P=0.001).Among patients who were positive for EGFR mutation, HLA-G 14 bp D/D genotype carriers had longer survival time than those carrying HLA-G 14 bp I/I or HLA-G 14 bp I/D genotype (P=0.041).The expression of HLA-G was closely correlated with HLA-G 14 bp polymorphism in patients with NSCLC (P=0.001).Conclusion These data, reported for the first time, indicates that HLA-G 14 bp polymorphism might be a genetic factor related to the susceptibility to NSCLC and associated with survival in patient with NSCLC after excluding the interference of molecular targeted agents.

Objective To evaluate the clinical significance of prenatal ultrasonography in diagnosis of retroaortic left innominate vein ( LINV ) . Methods Thirty fetus with retroaortic LINV were involved , including 17 cases of isolated retroaortic LINV and 13 cases of complicated retroaortic LINV . Three-vessel and trachea ( 3VT ) view was focused in the routine second trimester ultrasound screening to observe whether LINV existing or not . And then the transducer was rotated to the fetal sagittal view to assess the relationship between LINV and aortic arch and the convergence of left subclavian vein and left internal jugular vein into the LINV . Results The characteristic feature of retroaortic LINV was the LINV converge into right superior vena cava ( SVC) under the aortic arch ,not above the aortic arch . The most important plane to detect the retroaortic LINV was the 3VT view ,which revealed the abnormal vein lying to the left of pulmonary artery . The abnormal vein then could be traced and revealed its connection with the right SVC beneath the aortic arch on the transverse plane inferior to the 3VT view and on the fetal sagittal plane . Conclusions The whole course of retroaortic LINV can be revealed by ultrasound prenatally ,which can not only account for the abnormal vein to the left of pulmonary artery on 3VT view ,but also play an important role in surgery decision and future cardiac intervention treatment .

Objective To evaluate the clinical feasibility of laparoscopic cool-tip radiofrequency ablation (LCRFA)for renal cell carcinoma. Methods Twelve selected cases of primary renal, ceil carcinoma underwent LCRFA. Of them, 4 cases of left renal carcinomas, 7 cases had right renal carci-nomas and 1 case had bilateral renal carcinomas. The maximum diameter of the tumors was 2.1-8. 5 era. Eleven cases were T, No M0 and the other one was T2 N0 M0. Results The mean operation time was 92±24 min, and the mean blood loss was 50±29 ml. None of the cases need blood transfusion post-operation. No laparoscopic operative complications were observed. Six weeks after operation, complete ablation was achieved in 12 lesions and partial ablation in 1, with a complete ablation rate of 92.3%(12/13). There was no statistic change of Hb. ESR. SCr and GFR after operations (P>0.05). Ten cases underwent CT examination 3 months after the treatment ,9 cases showed complete necrosis of tumor,1 case showed partial necrosis. No recurrence was found. All of the 12 cases remained sur-vived during the follow-up for 1-16 months(median,7.8 months). Conelusions LCRFA for renal cell carcinoma is an accurate and effective intervention with a low incidence of complications, and is more accurate than ultrasound-guided pereutaneous radiofrequency ablation.

Objective To study the effect of flupentioxl melitracen and pinaverium bromide treatment on the changes of anorectal motility and rectal sensation in the patients of diarrhea-predominant irritable bowel syndrome (IBS-D) accompanying with depression and/or anxiety status. Methods Forty-four patients with IBS-D accompanying with depression and/or anxiety status were divided into group A (flupentioxl melitracen and pinaverium bromide) and group B (pinaverium bromide) by random digits table,and treated for 4 weeks. Twenty-five healthy subjects were included as control group. The anorectal motility and rectal sensation before and after taking medicines were compared. Results When abdominal pressure was increased, the net increased pressure of anal sphincter was (3.0 ± 1.2 ) kPa in group A and (2.9 ± 1.2)kPa in group B. They were lower than that in control group [(3.6 ± 1.6) kPa](P＜ 0.05). The rectal lowest volume of sensory threshold, the maximal volume of tolerance and maximal compliance were (55 ± 20) ml,( 145 ± 78 ) ml, ( 21.9 ± 12.9 ) ml/kPa in group A, ( 56 ± 38 ) ml, ( 150 ± 50 ) ml, (20.8 ± 11.2) ml/kPa in group B. They were lower than those in control group [(80 ± 38 ) ml, ( 190 ± 50 ) ml, (30.8 ± 15.2 ) ml/kPa](P ＜ 0.01 ). The rectal lowest volume of sensory threshold, the maximal volume of tolerance and maximal compliance were higher than those before taking medicines. Only the rectal lowest volume of sensory threshold in group B was higher than that before taking medicines. The rectal lowest volume of sensory threshold, the maximal volume of tolerance and maximal compliance in group A after taking medicines were higher than those in group B (P ＜ 0.05 or ＜ 0.01 ). Conclusions Higher sensitivity, lower tolerance,lower compliance of rectum and weakened anal automatic control function in IBS-D may be associated with diarrhea and frequent defecation. Treatment combining flupentioxl melitracen with pinaverium bromide may preferably improve the aperception functions of rectum in the patients of IBS-D accompanying with depression and/or anxiety status.