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

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

Objective To set up a method for identification of Lonicerae japonicae flos volatile oils using Fourier-transform infrared spectroscopy. Methods The volatile oils of Lonicerae japonicae flos and Lonicerae flos was extracted by steam distillation combined with continuous liquid-liquid extraction with hexane. An oil film was prepared for Fourier-transform infrared spectroscopy scanning by dropping the volatile oils solution on the KBr disc and evaporating the solvent. The obtained infrared spectrum was treated by baseline removing and median filter smoothing. The spectral data within 1800-850 cm-1 was selected as the characteristic spectrum for hierarchical cluster analysis. And the volatile oils of Lonicerae japonicae flos and Lonicerae flos were discriminated by the result of hierarchical cluster analysis. Results Enough volatile oils were extracted for obtaining Fourier-transform infrared spectrum from small amount of Lonicerae japonicae flos. The method developed in the study was able to discriminate Lonicerae japonicae flos volatile oils from Lonicerae flos volatile oils. Conclusion The method can be used for identification of Lonicerae japonicae flos volatile oils.

Objective To investigate the effects of testosterone (T) replacement therapy (TRT) on carotid artery intima-media thickness (IMT) in middle aged and elderly male patients.Methods A total of 80 middle-aged and elderly male patients with testosterone deficiency and increased carotid artery IMT were selected and randomly divided into two groups:the treatment group (n=38,treated with testosterone for 1 year) and the control group (n=42,without any treatment).The serum T level,IMT and prostate-specific antigen (PSA) before and after treatment were determined.The correlation between the testosterone level and carotid artery IMT was analyzed.Results There were no significant differences in the serum T level and IMT between the control group and the treatment group before treatment [(10.39 ± 1.44) nmol/L vs.(10.88 ± 1.87) nmol/L,(1.25 ±0.11) mm vs.(1.24±0.13) mm,t=1.32,-0.26,P=0.191,0.794].Compared with pretreatment,the serum T level was significantly increased and the IMT was significantly decreased in the treatment group afterTRT [(10.88±1.87) nmol/L vs.(22.83±1.56) nmol/L,(1.24±0.13) mmvs.(1.18±0.16) mm,t=-29.14,2.55,P=0.000,0.015],while no significant differences in the serum T level and IMT were found in the control group before and after treatment [(10.39± 1.44)nmol/L vs.(9.99±1.72) nmol/L,(1.25±0.11) mm vs.(1.27±0.11) mm,t=1.24,-1.00,P =0.219,0.323].Linear correlation analysis showed that the serum T level was negatively correlated with IMT (r-0.605,P=0.000) and multiple regression analysis showed that the T level was an independent factor for IMT.Conclusions Testosterone replacement therapy is an effective treatment to alleviate IMT in middle-aged and elderly male patients,which may play an important role in preventing cardiovascular diseases in middle-aged and elderly male patients.

OBJECTIVE:To compare the performance of several pattern recognition methods in the identification of volatile oils of traditional Chinese medicine(TCM)by infrared spectroscopy. METHODS:The volatile oils of several Lonicera and Citrus TCM were determined by infrared spectroscopy. All samples of infrared spectrum were classified by hierarchical clustering,K-mean clustering,artificial neural networks,and support vector machine. RESULTS:The results of hierarchical clustering and K-mean clus-tering were ineffective. Methods of artificial neural networks and support vector machine achieved correct classification rate of 100%. CONCLUSIONS:Artificial neural networks and support vector machine can be combined with infrared spectroscopy to cre-ate chemometric fingerprinting for the identification of volatile oils of TCM.

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.