Objective To investigate the influence of β-catenin gene deletion on Stat-5α phosphorylation in bcr-abl induced leukemia cells. Methods The established conditonal hematopoitic β-catenin knockout mice were used to isolate bone marrow cells. Exogenous bcr-abl fusion gene was transduced to these bone marrow cells by retroviral infection with intent to transfom them to leukemia cells.Immunofluorescence was performed to detect the phosphorylation status of Stat-5α in both β-catenin deletion cells and control cells. bcr-abl transcription and protein levels were evaluated with real-time PCR and western blotting. Results Phosphorylation of Stat-5α was reduced significantly in β-catenin deletion leukemia cells on comparison with control cells despite that total Stat-5α protein showed no obvious changes. Total tyrosine phosphorylation and bcr-abl protein expression were reduced in bcr-abl induced β-catenin deletion CML cells,on the contrary, both of the reduction were not seen in bcr-abl induced β-catenin deletion ALL cells.Conclusion Loss of β-catenin inhibits both Stat-5α phosphorylationin and bcr-abl expression in bcr-abl induced leukemia cells.

Arsenic trioxide albumin microspheres (As2O3-BSA-NS) were prepared by using methods of chemical cross-linking. The desirability function (DF), calculated according to the size (<1 microm) distribution, drug loading and drug trapping efficiency, was introduced as a total index for the microspheres formulation. Four factors, inculding W/O ratio, decentralization speed, BSA concentration and stirring stabilization time, were selected and arranged in an orthogonal experimental table. The release characteristic was studied by the drug release experiment in vitro. The four factors affected DF differently. Decentralization speed behaved as the maximum (P<0.01), followed by BSA concentration (P<0.05) and the W/O ratio dose (P<0.05). Stirring stabilization time did not influence DF (P>0.05). The release experiment in vitro showed that As2O3 in As2O3-BSA-NS was released more slower than pure As2O3. It was concluded that regular As2O3-BSA-NS may be prepared by the methods of chemical cross-linking, which was optimized by orthogonal experimental analysis of different factors, and the microspheres can release As2O3 slowly.
Arsenicals/*chemistry ; Cross-Linking Reagents/pharmacology ; Delayed-Action Preparations ; Drug Carriers/*chemistry ; Drug Delivery Systems ; Microspheres ; Oxides/*chemistry ; Serum Albumin, Bovine/*chemistry ; Technology, Pharmaceutical

ObjectiveTo observe the influence of medical exercise on rehabilitation of patients suffering from chronic obstructive pulmonary disease (COPD).Methods26 COPD patients who were organized in the form of training class practised medical exercise for three months. Respiratory function, exercise capacity and psychological health level were assessed respectively before and after exercise.ResultsValues of forced vital capacity(FVC), forced expiratory volume in one second (FEV1), FVC% , FEV1/ FVC and 6-minute walking distance (6MD) were significantly higher after medical exercise than before(P<0.01). Anxiety and depression scores also notably dropped after medical exercise than before(P<0.01).Conclusions Medical exercise can remarkably improve respiratory and moving capacity and decrease anxiety and depression of patients suffering from chronic obstructive pulmonary disease.

Objective:To investigate the influence of P27RF-Rho mRNA gene silencing in the drug sensitivity of 5-fluorouracil(5-Fu)to the liver cancer SMMC cell line,and to provide theoretical basis for the treatment of advanced liver cancer.Methods:The P27RF-Rho RNAi vector was constructed and the P27RF-Rho gene silencing lentivirus were used to infect the SMMC7721 cells.Western blotting method was used to detect the gene silencing effect.The SMMC7721 cells were divided into Scramble-siRNA group, 5-Fu group, P27RF-Rho siRNA group and P27RF-Rho siRNA + 5-Fu group.Western blotting was used to detect the transfection efficiency of RNAi.MTT method was used to detect the cell growth in various groups.Scratching test was used to detect the migration ability of cells in various groups.Transwell experiment were used to detect the invasion ability of cells in various groups.The expressions of P27 and RhoC protein were detected by Western blotting method.Results:P27RF-Rho RNAi lentiviral vector was successfully constructed.The Western blotting results showed that the expression of P27RF-Rho protein in P27RF-Rho siRNA group was decreased compared with 5-Fu group and Scramble-siRNA group(P<0.05).Compared with other three groups, the growth speed of the cells in P27RF-Rho siRNA + 5-Fu group was significantly decreased(P<0.05).The migration ability of the cells in P27RF-Rho siRNA + 5-Fu group was significantly lower than those in other three groups (P<0.01);the average number of cells passing through the Transwell microporous membrane was significantly less than those in other three groups (P<0.01).The Western blotting analysis results showed that the expression level of P27 protein in the cells in P27RF-Rho siRNA + 5-Fu group was significantly higher than those in other three groups(P<0.05);the expression level of RhoC protein was significantly lower than those in other three groups(P<0.05).Conclusion:P27RF-Rho gene silencing can significantly enhance the drug sensitivity of 5-Fu to SMMC7721 cells.

The biotransformation, CYP reaction phenotyping, the impact of CYP inhibitors and enzyme kinetics of 3-cyanomethyl-4-methyl-DCK (CMDCK), a new anti-HIV preclinical candidate belonging to DCK analogs, were investigated in human intestinal microsomes and recombinant cytochrome P450 (CYP) enzymes. CMDCK (4 micromol L(-1)) was incubated with a panel of rCYP enzymes (CYP1A2, 2C9, 2C19, 2D6 and 3A4) in vitro. The remaining parent drug in incubates was quantitatively analyzed by a LC-MS method. CYP3A4 was identified as the principal CYP isoenzyme responsible for its metabolism in intestinal microsomes. The major metabolic pathway of CMDCK was oxidation and a number of oxidative metabolites were screened with LC-MS. The Km, Vmax, CLint and T1/2 of CMDCK obtained from human intestinal microsome were 45.6 micromol L(-1), 0.33 micromol L(-1) min(-1), 12.1 mL min(-1) kg(-1) and 25.7 min, respectively. Intestinal clearance of CMDCK was estimated from in vitro data to be 3.3 mL min(-1) kg(-1), and was almost equal to the intestinal blood flow rate (4.6 mL min(-1) kg(-1)). The selective CYP3A4 inhibitors, ketoconazole, troleandomycin and ritonavir demonstrated significant inhibitory effects on CMDCK intestinal metabolism, which suggested that co-administration of CMDCK with potent CYP3A inhibitors, such as ritonavir, might decrease its intestinal metabolic clearance and subsequently improve its bioavailability in body.

Objective:To investigate the silencing of P27RF-Rho gene with lenvirus targeting mediated technique,and to clarify its influence in the invasion of liver cancer cells.Methods:The P27RF-Rho RNAi lentivirus was constructed. The liver cancer BEL7402 cells were infected with lentivirus. The experiment was divided into P27RF Rho-siRNA group, scramble-siRNA group and BEL7402 group.The effect of silencing P27RF-Rho gene and the expression levels of hepatocellular carcinoma (HCC)associated proteins RhoA,RhoC, VEGF,P53 and PTEN were detected;the activities of matrix metalloproteinase (MMPs)associated with tumor invasion were analyzed by Gelatin zymography;the variation of transfer ability and invasion abilities were compared by Wound healing assay experiment and Transwell experiment.Results:The Western blotting results showed the expression levels of P27RF-Rho,RhoA,RhoC,and VEGF proteins in the BEL7402 cells in experiment group were significantly lower than those in two control groups (P<0.05),and the expression levels of P53 and PTEN were higher than those in two control groups (P<0.05).The results of Gelatin zymography showed the activities of MMPs in experiment group were significantly lower than those in two control groups (P<0.01 );Wound healing assay showed that the migration ability of the BEL7402 cells in experiment group was significantly inhibited (P<0.01);the number of cells passed through the Transwell Chambers in experiment group was significantly less than those in two control groups (P<0.01).Conclusion:Silenceing P27RF-Rho can weaken the invasion ability and migration ability of human HCC BEL7402 cells.

Summary: Arsenic trioxide albumin microspheres (As2O3-BSA-NS) were prepared by using methods of chemical cross-linking. The desirability function (DF), calculated according to the size (<1 μm) distribution, drug loading and drug trapping efficiency, was introduced as a total index for the microspheres formulation. Four factors, inculding W/O ratio, decentralization speed, BSA concentration and stirring stabilization time, were selected and arranged in an orthogonal experimental table. The release characteristic was studied by the drug release experiment in vitro. The four factors affected DF differently. Decentralization speed behaved as the maximum (P<0.01), followed by BSA concentration (P<0.05) and the W/O ratio dose (P<0.05). Stirring stabilization time did not influence DF (P>0.05). The release experiment in vitro showed that As2O3 in As2O3-BSA-NS was released more slower than pure As2O3. It was concluded that regular As2O3-BSA-NS may be prepared by the methods of chemical cross-linking, which was optimized by orthogonal experimental analysis of different factors, and the microspheres can release As2O3 slowly.

Objective: To explore the correlation of non-coding RNA and the tumor-associated antigen midkine (MK) in SKOV3cells and the clinical significance for diagnosis of ovarian cancer. Methods: The Agilent′s gene chips (miRNAs chip and lncRNAs chip) were used to analyze the differential expression of miRNAs and lncRNAs in both MK-overexpressing SKOV3-MK cells and the control SKOV3-Con cells to screen the potential biomarkers in ovarian cancer. The clinical significance of midkine in the serum and tissues samples was analyzed for the patients with ovarian cancer by quantitative PCR combined with clinical data. Results: Compared with control SKOV3-con cells, MK overexpression significantly promoted the expressions of 11 miRNAs and 7 lncRNAs in SKOV3 cells (P＜0.01, ratio＞3 fold), reduced the expressions of 8 miRNAs and 13 lncRNAs (P＜0.01, ratio＜0.3). Results of qPCR showed that the expression level of miR489 was significantly lower in ovarian cancer tissues than that of the contralateral normal ovarian tissues, while HOTAIR was significantly elevated (P＜0.05). The expression level of HOTAIR in the serum of ovarian cancer patients was significantly higher than that in healthy controls group with same age (0.036±0.024 vs 0.019±0.020, P=0.002). ROC curve analysis of HOTAIR showed that the specificity was 66.7%, the sensitivity was 75.6% and the AUC value was 0.749 as a marker for serum detection of ovarian cancer when the cutoff value was 0.017 6. Conclusion Long-chain non-coding RNA HOTAIR may be served as a potential biomarker in serum of ovarian cancer patients.

Objective:Based on the cost-effectiveness,it aimed to assess the health benefits amd economic value of screening,di-agnosis,treatment,and optimal delicacy management of liver disease in hepatocellular carcinoma(HCC)patients.Methods:A Deci-sion tree-Markov model was developed to compare the cost-effectiveness of HCC screening and long-term surveillance versus no screening in population at risk from the health care system perspective.Results:It is found that HCC screening was a cost-effective approach compared to no screening(Incremental Cost-Effectiveness Ratio[ICER]:17 790 yuan/QALY).Scenario analyses suggested that initiating HCC screening at the age of 40,as recommended by clinical guidelines,and implementing long-term surveillance based on risk stratification were more cost-effective.Conclusions:For the implementation of HCC screening programs,attention should be paid to improving participation and compliance among the population at risk,incorporating advanced screening methods,improving management efficiency with digital tools,and introducing innovative payment methods to reduce economic burden.

Objective: To investigate the expression levels of long non-coding RNA LincROR in plasma and tissues of ovarian cancer patients and its value in the screening of ovarian cancer. Methods: The plasma samples from 30 healthy women, 56 cases of ovarian cysts, 23 cases of endometriosis, 38 cases of endometrial carcinoma, 35 cases of cervical cancer, 42 cases of ovarian cancer, 21 cases of ovarian cancer after operation and 26 cases of ovarian cancer after chemotherapy were collected, and the expression levels of LincROR in these samples were detected by quantitative PCR. The diagnostic value of LincROR in common clinical gynecological diseases was evaluated combined with clinical data. Results: The expression levels of LincROR in plasma of ovarian cancer patients (2.90± 4.42 ) were significantly higher than that in healthy women (0.23±0.28) and the patients with benign ovarian cysts (0.62±0.55, P ＜ 0.01 ). The results of ROC curve analysis showed that the diagnostic value of plasma LincROR in the screening of breast cancer was better than that of CA125, CA199, CA153, AFP and CEA. The sensitivity and specificity of combined screening of LincROR and CA125 for ovarian cancer were 89.7% and 86.7%, respectively (AUCROC=0.918, 95% CI :0.817-0.973). In addition, the expression levels of plasma LincROR in the postoperative patients were significantly lower than that in the ovarian cancer patients without any treatment (0.50±1.72 vs 2.90±4.42, P ＜0.01). The ROC curve analysis showed that plasma LincROR was more sensitive than CA125 in evaluating the efficacy of chemotherapy for ovarian cancer (AUCROC: 0.866 vs 0.738). Conclusion LincROR is expected to be an ideal biomarker for the screening of ovarian cancer, and has potential clinical value in evaluating the efficacy of chemotherapy for ovarian cancer. Combination of LincROR with CA125 may improve the sensitivity and specificity of the screening of ovarian cancer