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.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; CD56 Antigen ; metabolism ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Fever of Unknown Origin ; etiology ; Humans ; Lymphoma, Extranodal NK-T-Cell ; complications ; metabolism ; pathology ; therapy ; Male ; Prednisone ; therapeutic use ; Splenectomy ; Splenic Neoplasms ; complications ; metabolism ; pathology ; therapy ; Vincristine ; therapeutic use ; Young Adult

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 conduct cloning, sequencing and expression of Trichinella spiralis ES antigen p49 gene. Methods RT-PCR was used to amplify the specific gene fragment from the total RNA of Trichinella spirais larvae. The PCR product was ligated to the T-vector and the recombinant plasmid was verified by sequencing. T-p49 and pGE-4T-3 were treated by both BamHI and XhoI. The ligation reaction was catalyzed by T4 DNA ligase. Results The p49 gene was cloned by using RT-PCR. Sequence analysis showed that the p49 gene obtained was consistent to the p49 sequence reported in the database. The expressed protein was shown as a new band at SDS-PAGE. BLAST analysis demonstrated that this p49 gene was 99% identical to the p49 gene reported and to the 43 kDa secreted glycoprotein gene in the database. Conclusion p49 gene from Trichinella spiralis larvae was cloned, sequenced and expressed.

Objective To investigate whether there existed a healthy obese subtype.Methods A total of 116 healthy subjects were recruited.They were divided into 3 groups according to BMI and metabolic disorders:40 cases of normal weight and metabolic normality (NMN),36 cases of obesity and metabolic normality (OMN) and 40 cases of obesity and metabolic abnormality (OMA).Anthropometic parameters as height,weight,waist circumference,hip circumference and blood pressure was recorded.Blood glucose,lipids,insulin,high-sensitivity C-reactive protein (hs-CRP) was detected.Body fat distribution was detected by dual-energy X-ray absorptiometry (DXA).Serum von Willebrand factor (vWF),a marker of endothelial dysfunction,were detected by ELISA.Results Both serum vWF levels in OMN group [(733.6±86.2)U/L] and OMA group[(809.2 ±46.3)U/L] are higher than that in NMN group [(466.9 ±65.3)U/L,P ＜0.05] with serum vWF level in OMA group is higher than in OMN group (P ＜ 0.05).Among android fat mass percentage (AFM％),BMI,waist height ratio,waist circumference,hs-CRP,weight,hip circumference and trunk fat mass,AFM％,BMI and hs-CRP are main influencing factors of vWF.Conclusions Endothelial dysfunction existed in obese adults regardless of their metabolic status.There is no healthy obese subtype.AFM％,BMI and hs-CRP are the main influencing factors of endothelial dysfunction.

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 evaluate the feasibility of an intelligent three-dimensional ultrasound technique (Smart mid-sagittal planes,Smart MSP) in automatically detecting the fetal cranial mid-sagittal view.Methods Two hundred and forty pregnant women with singleton pregnancies were imaged to display the mid-sagittal view of fetal head using the Smart MSP by six doctors divided into three groups according the different experiences.Another two doctors were then invited to score the images acquired and to compare the successful rates of 6 doctors for acquiring fetal cranial mild-sagittal view according to the doctors' experience.Results The overall successful rate of acquiring the mid-sagittal view was 97.08％ (233/240) with the bipartial diameter plane (BPD plane) as the starting plane and 98.33(237/240) with the trans-cerebellum plane as the starting plane by using Smart MSP program.The successful rates in three groups were similar but the consumed times were different(P ＜0.05).The experienced doctors consumed less time than non-experienced doctors.The scores were also different in three groups.The scores of experienced doctors (group B and C) were significantly higher than that of non-experienced doctors (Group A) (P ＜0.05).There was no significant difference in the scores between experienced doctors (group B and C) (P＞0.05).Conclusions Smart MSP is a novel and feasible method for the automatic visualization of fetal cranial mid-sagittal plane,which may become an effective tool to screen the fetal midline anomalies in future.