Objective To investigate the expression level of miR-133b in cancer tissues of patients with prostate cancer and its effect on the proliferation of prostate cancer cells.Methods The total RNAs in resected prostate cancer tissues and adjacent tissues from 30 patients with prostate cancer were extracted and reversely transcripted into cDNA,and then the expression levels of miR-133b were detected by real-time quantitative PCR.The correlations between the expression levels of miR-133b and the patients' clinicopathological features were analyzed.The expression of positive regulatory domain I-binding factor 16 (PRDM16) and proliferation of PC-3 cells transfected with miR-133b mimics by LipofectamineTM 3000 were determined by real-time quantitative PCR and the CCK8 method,respectively.Results The expression levels of miR-133b in prostate cancer tissues [(16.85 ± 0.94) × 10-4] was significantly lower than that in adjacent tissues [(22.95 ± 1.567) × 10-4,t =3.335,P ＜ 0.01].The expression levels of PRDM16 in PC-3 cells transfected with miR-133b mimics were significantly lower than that in the control group (0.371 ±0.031 vs 1.000 ±0.022,t =12.53,P ＜ 0.01).The proliferation ability of PC3 cells transfected with miR-133b mimics for 72 hours was significantly lower than that in the control group (t =6.811,P ＜ 0.01).Similarly,the proliferation ability of PC-3 cells transfected with PRDM16 inhibitor for 72 hours was also significantly lower than that in the control group (t =9.048,P ＜0.01).Conclusion The expression levels of miR-133b in prostate cancer tissues are significantly down-regulated,which regulate the proliferation of prostate cancer cells possibly through PRDM16.

Objective To investigate the significance of miR-103 as noninvasive biomarker for diagnosis of nonalcoholic fatty liver disease (NAFLD).Methods The serum samples were collected from healthy subjects and NALFD patients to detect biochemical parameters.NucleoZOL method was used to isolate serum miRNA.SYBR Green qPCR was used for relatively quantitative analysis of miR103a-2.The correlations between miR-103a-2 and biochemical parameters were analyzed by Spearman method.Results Compared with healthy control group,the levels of total cholesterol (TC),triacylglyceride (TG),alanine aminotransferase (ALT) and glutamyl transpeptidase (γ-GGT) were all significantly increased in NAFLD group (P ＜ 0.01).Compared with the NAFLD group accompanying ortholiposis,the levels of TC,TG and low density lipoprotein-cholesterol (LDL-C) were all significantly increased in the NAFLD group accompanying dyslipidemia (P ＜0.01).Compared with the healthy control group,the level of serum miR-103a-2 increased in both NAFLD with ortholiposis group (P ＜ 0.05) and NAFLD with dyslipidemia group (P ＜ 0.01).The level of serum miR-103a-2 was positively correlated with serum TC (r =0.495,P =0.001).Conclusion Serum miR-103a-2 may be a more sensitive parameter for noninvasive screening of NAFLD than the parameters of blood lipid.

Objective To investigate the diagnostic value of seven tumour-associated autoantibodies,including p53,PGP9.5,SOX2,GAGE7,GBU4-5,MAGE A1 and CAGE autoantibodies,in the newly diagnosed patients with lung cancer.Methods A total of 108 patients with newly diagnosed lung cancer,120 with benign lung diseases,227 with other cancers and 96 healthy controls were enrolled in the study.Their serum levels of seven autoantibodies were detected by enzyme linked immunosorbent assay (ELISA).The ROC curve was drawn and used to analyze their diagnostic efficiency for lung cancer.The diagnostic value of the combination of seven autoantibodies in different groups was also compared.Results The serum levels of seven autoantibodies in lung cancer patients were significantly higher than those in the patients with benign lung diseases or other cancers and healthy controls (P ＜ 0.05).The sensitivity,specificity and AUCROc of the combination of seven autoantibodies in the preliminary diagnosis of lung cancer were 62.00％,89.80％ and 0.769,respectively,and its sensitivity and AUCROC were higher than those of single autoantibody.The positive rate of the combination of seven autoantibodies in lung cancer patients was significantly higher than those in healthy controls (x2 =50.885,P ＜ 0.01) and the patients with benign lung diseases (x2 =56.341,P ＜ 0.01) or other cancers (x2 =46.812,P ＜ 0.01).Conclusion The combination detection of seven autoantibodies,including p53,PGP9.5,SOX2,GAGE7,GBU4-5,MAGE A1 and CAGE,may serve as potential markers for the diagnosis of lung cancer.

Objective To investigate the cellular immunologic response of TH 17/Treg cells in the peripheral blood of pelvic tuberculosis patients and explore their roles in the pathogenesis of pelvic tuberculosis.Methods The intracellular flow cytometry was performed to evaluate the expressions of TH 17 and Treg cells in 46 pelvic tuberculosis patients and 25 healthy controls in childbearing age.Twenty-eight of the 46 pelvic tuberculosis patients were followed up to monitor the variation of the TH17/Treg cells after 3 months and 6 months of anti-tuberculosis treatment.Results The percentage of TH 17 cells in the peripheral blood of pelvic tuberculosis patients was (3.26 ± 1.30) ％ which was significantly lower than that of healthy controls [(4.92 ± 1.71) ％,P ＜ 0.01].The percentage of Treg cells in the patients was (5.18 ± 1.53) ％ which was significantly higher than that of healthy controls [(3.26 ± 1.10) ％,P ＜ 0.01].The percentage of TH17 cells in the pelvic tuberculosis patients after 6 months of treatment was (4.67 ± 1.75) ％ which was significantly higher than that in the patients before treatment and after 3 months treatment [(3.26 ± 1.30) ％,P ＜ 0.01 and (3.70 ± 1.06) ％,P ＜0.01,respectively].The percentage of Treg cells in pelvic tuberculosis patient after 6 months of treatment was (3.93 ±0.94)％ which was significantly lower than that in the patients before treatment and after 3 months of treatment [(5.18 ± 1.53)％,P ＜0.01 and (4.94 ± 1.51) ％,P ＜ 0.01,respectively].The percentage of Treg cells in the patients after 6 months of treatment was still significantly higher than that of controls (P ＜ 0.05).The TH 17/Treg ratio before treatment was significantly lower than that of healthy controls (P ＜ 0.01),and the TH 17/Treg ratio was increased after 3 months of treatment but it did not show significant difference compared with that before treatment.The TH 17/Treg ratio after 6 months of treatment (1.18 ± 0.34) ％ was significantly increased in contrast to those after 3 months of treatment and before treatment [(0.77 ± 0.21) ％,P ＜ 0.01 and (0.55 ± 0.13) ％,P ＜ 0.01,respectively].The TH 17/Treg ratio could not rise to the normal level even after 6 months of treatment.Conclusion Both the TH 17 and Treg cells may involve in the immunologic responses of pelvic tuberculosis patients and the imbalance of TH1T/Treg cells may remain persistently.

Objective To analyze the mutation characteristics of GBA gene in one patient with Gaucher disease and platelet transfusion refractoriness.Methods A female patient with anemia and thrombocytopenia showed platelet transfusion refractoriness,and then the proband and her family were performed bone marrow smear,β-glucocerebrosidase activity in leukocytes (dried blood spot assay),Bultrasonography and gene sequencing examination and pedigree investigation.Results Pedigree investigation showed that the heterozygous mutation of GBA gene existed in the father,mother,son,daughter and sister of the proband.Bone marrow cytomorphologic examination showed that Gaucher cells accounted for 6.0％ in the female patient.The β-glucocerebrosidase activity in leukocytes was 3.78 nmol/(h · mg Pro).B-ultrasonography showed slightly splenomegaly.Gene sequencing found that the homozygous mutation of GBA gene,c.484A ＞ G,existed in the female patient.Conclusion The patients with Gaucher disease may appear platelet transfusion refractoriness due to hypersplenism.The mutation of GBA gene is the main pathogenic factor of the family with Gaucher disease.

Objective To carry out a systematic evaluation for the diagnostic accuracy of Des-γ-carboxy-prothrombin(DCP) in primary hepatocellular cancer(PHC) by Meta-analysis.Methods The published international and domestic studies on DCP in the diagnosis of PHC were searched in multiple databases from its inception until December 2016.A total of seventeen studies were finally selected,among which 1 970 cases of PHC group and 2 588 cases of control group were included.The control group was composed of chronic hepatitis,cirrhosis,tumors in other systems and healthy subjects with physical examination.A meta-analysis was performed using Stata 12.0.Results The overall pooled results of the analysis and the 95％ confidence intervals were as follows:the sensitivity(SEN) was 0.79 (0.74 to 0.83),the specificity (SPE) was 0.90 (0.87 to 0.93),the positive likelihood ratio(PLR) was 8.1 (5.8 to 11.1),the negative likelihood ratio(NLR) was 0.24(0.19 to 0.29),the diagnostic odds ratio(DOR) was 34(21 to 55) and the area under the summary receiver operator characteristic curve (SROC) was 0.91 (0.89 to 0.93).The sensitivity combined the detections of AFP and DCP in the diagnosis of PHC was 0.90 with 0.94 of AUCROC and 56 of DOR.Conclusion The serum DCP may exhibit a relatively higher diagnostic specificity for PHC.The combined detections of AFP and DCP could improve the sensitivity and diagnostic accuracy.

Objective To investigate the effects of liver X receptor (LXR) agonist on the proliferation of mouse pancreatic β cell line MIN6 cells.Methods The viability,changes of cell cycle,mRNA levels of S phase kinase associated protein 2 (Skp2) and p27,and protein levels of Skp2 and p27 in MIN6 cells treated with LXR agonist T0901317 were determined by the CCK-8 method,flow cytometry,real-time RT-PCR and western blot,respectively.Results The viability of MIN6 cells treated with 1 μmol/L,5 μmol/L and 10 μnol/L of T0901317 were (98.54 ±0.94)％,(87.03 ±0.93)％ and (75.57 ± 1.85)％ of the controls,respectively,and there was significant difference among them (F =301.90,P ＜ 0.01).The percentages of G1 phase cells in the MIN6 cells treated with 0 μmol/L,1 μmol/L,5 μmol/L and 10 μmol/L of T0901317 were (35.93 ±2.25)％,(38.45 ±0.91)％,(45.46±1.34)％ and (53.28 ± 1.14) ％,respectively,and there was significant difference among them (F =80.83,P ＜ 0.01).Similarly,the percentages of S phase cells in the MIN6 cells treated with 0 μmol/L,1 μmol/L,5 μmol/L and 10 μmoi/L of T0901317 were (52.87 ± 1.19) ％,(48.65 ± 0.85) ％,(36.31 ± 1.37) ％ and (31.45 ± 1.22) ％,respectively,and there was also significant difference among them (F =221.30,P ＜ 0.01).The protein levels of p27 in the MIN6 cells treated with 10 μmol/L of T0901317 (2.84 ± 0.14) were significantly higher than that in the controls (2.28 ± 0.10) (t =4.54,P ＜ 0.05),while there was no significant difference in the mRNA levels of p27 between them (t =0.28,P ＞ 0.05).However,10 μmol/L of T0901317 significantly decreased mRNA (0.52 ± 0.02,t =29.22,P ＜ 0.01) and protein levels (0.98 ± 0.12 vs 1.89 ± 0.01,t =10.98,P ＜ 0.01) of Skp2 in MIN6 cells.Based on the control siRNA transfection group as a reference (100％),the cell survival rates of the p27 siRNA transfection group,10 μmol/L of T0901317 treatment group and the intervention group (p27 siRNA transfection + T0901317 treatment) were (100.97 ± 1.08) ％,(75.03 ± 1.83) ％ and (86.67 ± 2.45) ％,respectively.There was no significant difference between the control siRNA and p27 siR-NA transfection groups (t =1.542,P ＞ 0.05).Compared with the control siRNA transfection group,the cell survival rates of the T0901317 treatment group decreased (t =23.58,P ＜ 0.01).There was also significant difference in the cell survival rates between the T0901317 treatment group and the intervention group (t =7.77,P ＜ 0.01).Conclusion The activation of LXR may induce pancreatic β cell cycle arrest by up-regulating the expression of p27 and down-regulating the expression of Skp2.

Objective To investigate the differences in the gene expression profiles between SW480 and SW620 cell lines.Methods A dataset of GDS756 containing the gene expression profiles of SW480 and SW620 was downloaded from the GEO database in NCBI.The differential expression genes between SW480 and SW620 were analyzed with gene set enrichment analysis (GSEA) and leading edge subset analysis.The genes in leading edge subset were re-annotated by FunRich software.The core genes of leading edge subset closely relating to SW480 or SW620 were analyzed with the STRING on-line analytical system.The functional core genes closely relating to SW480 or SW620 were obtained by the combined analysis of the core genes and high frequency genes from leading edge subset.Results GSEA identified 12 significantly enriched gene sets,491 leading edge genes and 7 highly overlapping genes from SW480 and 80 significantly enriched gene sets,870 leading edge genes and 6 highly overlapping genes from SW620.The STRING system identified 5 core genes from SW480 and 8 from SW620.The combined analysis of GSEA and bionetwork obtained 2 functional core genes,TOP2A and CDK1,from SW620.Conclusion The SW480 and SW620 cells with identical genetic background have different functional gene expression profiles,and the functional core genes TOP2A and CDK1 in SW620 cells may be related to the signal pathways of colon cancer metastasis.

Objective To explore the effects of all-trans-retinoic acid (ATRA) on the proliferation of human lung adenocarcinoma cell line A549 and the expression of APLNR (apelin receptor) gene.Methods The inhibition of proliferation of human lung adenocarcinoma cell lines A549 cultured in vitro with or without ATRA was measured by MTT (methyl thiazolyl tetrazolium,MTT) method.The morphological changes in the cells were observed by light microscopy.The cell cycle and apoptosis were analyzed by flow cytometry.The levels of APLNR,cyclin D1 and p16 proteins were detected by western blot.Results After treatment of ATRA,the proliferation of A549 cells was obviously inhibited in dose-and time-independent manner (P ＜ 0.01).The cell morphology was significantly changed.The cycle of A549 cells was blocked at G0/G1 phase and the apoptosis rate was increased.With the increasing concentration of ATRA,the expressions of cyclin D1 and APLNR were down-regulated but the expression of p16 was up-regulated (P ＜ 0.01).Conclusion ATRA could inhibit the proliferation of A549 cells by retardant cell cycle of A549 cells at G0/G1 phase and inducing the apoptosis,and down-regulate the expression of APLNR gene.

Objective To investigate the effects of down-regulation of lemur tyrosine kinase 3 (LMTK3) on the cytobiological behaviors of human lung cancer A549 cells.Methods The expression of LMTK3 in A549 cells was interfered with small hair RNA (shR-NA),and the expression level of LMTK3 was determined by RT-PCR.The effects of LMTK3 on the proliferation,migration,cell cycle and apoptosis of A549 cells were determined by the CCK-8 assay,scratch assay,Transwell assay and flow cytometry,respectively.Results The shLMTK3 cells with stably low expression of LMTK3 and control cells (Scramble) with normal expression of LMTK3 were successfully obtained.The relative proliferation rates of shLMTK3 cells cultured for 24,48 and 72 hours were significantly lower than those in the control cells (0.305 ±0.018 vs 0.354 ±0.011,t =5.24,P＜0.01;0.461 ±0.044 vs 0.551 ±0.027,t =3.91,P ＜0.01;0.74 ± 0.029 vs 0.881 ± 0.028,t =7.70,P ＜ 0.01).The relative migration rate of shLMTK3 cells 24 hours after scratching was significantly lower than that of control cells (0.51 ±0.096 vs 1.00 ± 0.029,t =4.81,P ＜ 0.01).Transwell assay showed that the number of migration cells in shLMTK3 group was significantly less than that in Scramble group (161 ±9.29 vs 308.66 ± 17.60,t =7.42,P ＜ 0.05).The results of flow cytometry showed that shLMTK3 cells were blocked at G1 phase (t =4.35,P ＜ 0.05),and that the inhibition of LMTK3 had no influence on the apoptosis of A549 cells.Conclusion Down-regulation of LMTK3 expression in human lung cancer A549 cells may inhibit the proliferation and migration of A549 cells significantly,indicating that the abnormal expression of LMTK3 in lung carcinoma cells may regulate the biological behaviors and progression of tumors.