Objective:To analyze the expression of long non-coding RNA (lncRNA) ZFPM2-AS1 in bladder cancer tissues and cell lines, and to observe the effect of down-regulating ZFPM2-AS1 on the migration and proliferation of bladder cancer cells and explore its molecular mechanism.Methods:Real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expression of ZFPM2-AS1 in 51 pairs of bladder cancer tissues and adjacent tissues, bladder cancer cell lines (J82, 5637, BIU-87, T24) and human normal bladder epithelial cells SV-HUC-1. The bladder cancer cells with the highest ZFPM2-AS1 expression were selected and transfected with the small interfering siRNA-ZFPM2-AS1 plasmid and the negative control plasmid, respectively, and defined as the experimental group and the control group. qRT-PCR was used to detect the expression of ZFPM2-AS1 in two groups of cells. Transwell migration test and tetramethylazozole blue (MTT) method were used to detect the cell migration ability and proliferation ability of the two groups. qRT-PCR was used to detect the expression of Up-frameshift mutant 1 (UPF1) mRNA in two groups of cells. Western blot was used to detect the expression of UPF1 and mTOR signaling pathway proteins in the two groups of cells.Results:The expression of ZFPM2-AS1 in bladder cancer tissues was significantly higher than that in adjacent tissues ( P<0.01). The expression of ZFPM2-AS1 in bladder cancer cell lines was significantly higher than that in human normal bladder epithelial cells ( P<0.01), and ZFPM2-AS1 had the highest expression in BIU-87 cells ( P<0.01). Compared with the control group, the expression of ZFPM2-AS1 in BIU-87 cells in the experimental group was significantly reduced [(1.01±0.06) vs (0.16±0.04), t=12.28, P<0.01]. Compared with the control group, the migration ability of BIU-87 cells in the experimental group was decreased ( P<0.05), and the proliferation ability of BIU-87 cells was significantly decreased from the second day ( P<0.05). Compared with the control group, UPF1 mRNA expression in BIU-87 cells in the experimental group was significantly decreased [(1.00±0.02) vs (0.28±0.04), t=15.49, P<0.01]. Western blot results showed that UPF1 protein expression and mammalian rapamycin target protein (mTOR), GRB2, IRS1 and p-PI3K signal pathway protein expression were decreased in BIU-87 cells. Conclusions:ZFPM2-AS1 is highly expressed in bladder cancer tissues and cell lines. Down-regulating ZFPM2-AS1 can inhibit the migration and proliferation of BIU-87 cells. The molecular mechanism may be related to the inhibition of UPF1 gene expression.

Objective:To observe the expression of microRNA (miRNA, miR) -7850 in renal cancer tissues, and to explore the effect of miR-7850 on the proliferation and migration of renal cancer cells and on the regulation of serine proteinase inhibitor B3 (SERPINB3) gene expression.Methods:Real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7850 in renal cancer tissues and renal cancer cell lines. The renal cell carcinoma cell line with the lowest expression of miR-7850 was selected, and the negative control sequence (miR-NC) and miR-7850 mimics were transfected into renal cell carcinoma cells by Lipofectamine 2000 transfection reagent, respectively, which were defined as miR-NC group and miR-7850 group. qRT-PCR was used to detect the expression of miR-7850 in transfected renal cancer cells. The cell proliferation and migration ability after transfection were detected by cell counting kit-8 (CCK-8) method and transwell experiment. Bioinformatics prediction and dual luciferase reporter gene experiments were used to verify the target gene of miR-7850. qRT-PCR and Western blot were used to detect the expression of target genes in renal cancer cells after transfection.Results:Compared with adjacent tissues (5.95±0.44), the expression of miR-7850 in kidney cancer tissues (1.19±0.33) was lower ( P<0.01). Compared with immortalized proximal renal tubular epithelial cells (1.01±0.07), the expression of miR-7850 was lower in renal cancer cell lines ( P<0.05), and the lowest in A498 cells (0.13±0.01) ( P<0.01). The expression of miR-7850 in the miR-7850 group (7.46±0.93) was significantly higher than that in the miR-NC group (1.01±0.08) ( P<0.01), indicating successful transfection. Compared with the miR-NC group, the cell proliferation ability of the miR-7850 group was significantly reduced ( P<0.05). The number of migrating cells in miR-NC group and miR-7850 group were (139.50±12.31) and (75.09±16.05) cells, respectively, and the cell migration ability in miR-7850 group decreased significantly ( P<0.01). Bioinformatics technology shows that the target gene of miR-7850 was SERPINB3. The dual luciferase reporter gene experiment confirmed that miR-7850 can target the SERPINB3 gene ( P<0.05). Compared with the miR-NC group, the expression of SERPINB3 in cells of miR-7850 group was significantly reduced ( P<0.05), as well as the CDK4, CyclinD, Snail and Vimentin. Conclusions:miR-7850 is lowly expressed in renal cancer tissues and cell lines. miR-7850 can inhibit the proliferation and migration of renal cancer A498 cells, which may be related to its inhibition of SERPINB3 gene expression.

OBJECTIVE To investigate the drug resistance of clinical isolates of Streptococcus pneumoniae and molecular epidemiology with BOX-PCR in Nanchang. METHODS Antibiotic susceptibility was determined by the Etest methods or K-B disk diffusion test. Twenty-nine Penicillin-resistant S. pneumoniae(PRSP) clinical isolates were molecularly typed by BOX-PCR to detect the clonal relationship of them. RESULTS 85.7% were resistant to erythromycin,57.1% were PRSP. The resistant rates to tetracycline,clindamycin,trimethoprim-sulfamethoxazole and chloramphenicol were 88.1%,79.8%,60.7%,and 28.6%. Amoxicillin/clavulanic acid,levofloxacin and ceftriaxone retained activity against most of the isolates. All of strains were susceptible to vancomycin. Twenty-nine S. pneumoniae strains were divided into 14 distinct types,and 19 subtypes with subtype A (n=4) as the predominant type. CONCLUSIONS The resistance rate of S. pneumoniae to penicillin is very high,BOX-PCR typing demonstrats a high discriminatory potential and easy to be accurately analyzed.

Objective To investigate the importance of plasmid-mediated quinolone resistance in the development of quinolone resistance in clinical isolates of gram-negative bacteria.Methods A total of 541 consecutive clinical isolates of gram-negative ba- cilli resistant or intermediate to ciprofloxacin were screened for the qnrA gene by PCR.Conjugation experiments were carried out with azide-resistant E.coli J53 as a recipient.The aac(6')-Ib-cr gene was detected.The mutations in the quinolone-resist- ance-determining region (QRDR) of the gyrA and parC genes were identified in qnrA positive strains.Results qnrA was identi- fied in 7 of the 541 strains.Among the qnrA positive strains,5 were Enterobacter cloacae.No qnrA was detected in nonfer- menters.Quinolone resistance was transferred in 4 of 7 qnrA positive strains.Transconjugants had 12-to 125-fold increases in MIC of ciprofloxacin relative to that of the recipient.Seven strains contained qnrA with a nucleotide sequence identical to that originally reported.Two transconjugants with higher ciprofloxacin MICs contained aac(6')-Ib-cr gene.Mutations occurred in the QRDR of the gyrA and parC genes in 5 PCR-positive clinical strains.Conclusions Transferable plasmid-mediated quinolone resistance associated with qnrA is highly prevalent in clinical strains of Enterobacter spp.aac(6')-Ib-cr gene and mutations in the quinolone targets may co-exist with qnrA,which may contribute to the further increase of resistance to quinolones.

Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

This paper, from the aspects of neuroendocrine, emotion, learning etc, analyzed comprehensively the characteristics of the influence of prenatal stress (PS) on offspring, including intensity of PS, timing of exposure and individual differences. Despite the variety in methodology, most studies on it indicate that the prenatal stressful events lead to offspring's increased plasma glucocorticoid (GC) level, more depressed-related behaviors and impaired learning abilities. Although mechanism of prenatal stress is still unclear, most studies show that it is concerned with hypothalamio-pituitary-adneral (HPA) axis,dopaminergic (DA-ergic) system, neuropeptide Y (NPY)and serotoninergic (5-HT ergic) system. Moreover, further studies are proposed to pay more attention to the relationship and interaction of various related substances.

To explore the expression of livin gene in acute non-lymphocytic leukemia (ANLL) cells and its clinical significance, the mRNA level of livin gene in 46 ANLL adult patients were measured by using reverse transcription polymerase chain reaction (RT-PCR). Other 10 healthy adults were selected as normal controls (NC), HL-60 cell line was employed as positive control. The results showed that the mRNA level of livin gene in ANLL patients was significantly higher than that in NC, while it decreased in patients with complete remission (CR). In relapsed patients, the level of livin mRNA increased again. In ANLL patients, the CR rate of patients with livin positive was lower than that of patients with livin negative (p<0.05). It is concluded that overexpression of livin gene may play a synergic role in the pathogenesis of ANLL and associates with CR rate in ANLL. It seems that high expression of livin gene may be used as a marker of poor prognosis in acute non-lymphocytic leukemia.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; genetics ; metabolism ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; Prognosis ; RNA, Messenger ; genetics ; metabolism ; Young Adult

Adult ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hepatitis B, Chronic ; genetics ; immunology ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic

OBJECTIVETo investigate the association between the polymorphism of human leucocyte antigen (HLA)-DRB1, -DQA1 and -DQB1 alleles and viral hepatitis B.

METHODSHLA-DRB1, -DQA1 and -DQB1 alleles in 52 patients with chronic hepatitis B, 30 patients with acute hepatitis B and 106 normal control subjects were analysed, using the polymerase chain reaction/sequence specific primer (PCR/SSP) technique.

RESULTSThe allele frequencies of HLA-DRB1 * 0301, -DQA1 * 0501 and -DQB1 * 0301 in the chronic hepatitis B group (17.31%, 25.96%, 35.58%) were markedly higher than that in the normal control group (5.67%, 13.36%, 18.87%), with statistical significance (chi(2)(1) = 12.3068, P(c1) = 0.0074; chi(2)(2) = 9.2002, P(c2) = 0.0157; chi(2)(3) = 15.5938, P(c3) = 0.0075). The allele frequencies of HLA-DRB1 * 1101/1104 and -DQA1 * 0301 in the chronic hepatitis B group (0.96%, 14.42%) were markedly lower than that in the acute hepatitis B group (13.33%, 30%), with significant correlation between them (chi(2)(1) = 11.9206, P(c1) = 0.0145; chi(2)(2) = 8.7396, P(c2) = 0.0167).

CONCLUSIONHLA-DRB1 * 0301, -DQA1 * 0501 and -DQB1 * 0301 were closely associated with the susceptibility to chronic hepatitis B, while HLA-DRB1 * 1101/1104 and -DQA1 * 0301 closely associated with the resistance to chronic hepatitis B. These findings suggested that host HLA class II gene was an important factor determining the outcome of HBV infection.

Adult ; Alleles ; DNA ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; HLA-DQ Antigens ; genetics ; HLA-DQ alpha-Chains ; HLA-DQ beta-Chains ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hepatitis B ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic

Objective To explore the effect of recovery sleep on the executive function after 36 h of total sleep deprivation by event related potential technology.Methods Thirteen healthy male college students participated in two trials. At the first trial normal sleep as control was investigated. At the second trial participants experienced 36 h of sleep deprivation and then accepted 8 h recovery sleep. In each trial six Go/Nogo tests were employed to test the executive control function and the ERP data were recorded. Results There was no statistical difference in behavior and ERP results at each time point as the subjects had normal sleep. After 36 h of sleep deprivation, the behavior results were statistically significant when compared to the baseline. The amplitude and latency of Nogo-N2, Nogo-P3 on Fz electrode, the amplitude and latency of Nogo-P3 on Cz electrode showed statistical significance when compared to the baseline. After 8 h recovery sleep, the average correct reaction time and the Go correct reaction rate had statistical significance compared to 36 h value. The amplitude of Nogo-N2 and Nogo-P3 had no statistical significance compared to the baseline.However,it was of statistical significance[(-6.80 3.95)vs(-3.37 2.63)μV,(10.63±6.62)vs(5.63±5.45)μV,(9.49±7.37)vs(6.08±6.56)μV] compared to 36 h value. The latency of the recovery value of Nogo-N2 and Nogo-P3 was statistically significant[(254.14±15.55)vs(243.08±13.97)ms(382.14±41.07)vs(349.17±30.36)ms,(369.86±26.48)vs(347.48±29.24)ms]compared to the baseline.Conclusion As the time of sleep deprivation is prolonged, the executive function is impaired and the executive function is not completely recovered after 8 h recovery sleep.