Objective To investigate the regulatory effects of miR-145 on the proliferation,cell cycle and apoptosis of a human keratinocyte cell line HaCaT.Methods miR-145 mimics and negative control (NC) mimics were chemically synthesized and then transiently transfected into HaCaT cells respectively.After additional culture for different durations,real-time PCR was performed to determine the expression level of miR-145,MTS assay to estimate cell proliferation,and flow cytometry to detect cell apoptosis and cycle.Luciferase assay,real-time PCR and Western blot were conducted to determine whether NRAS was the target gene of miR-145.Results The miR-145 expression level in miR-145 mimic-transfected cells increased by 85.00 ± 1.21 folds compared with NC mimic-transfected cells (t =115.90,P ＜ 0.0001).The transfection with miR-145 mimics significantly inhibited the proliferation of HaCaT cells (F =8.76,P =0.008),and the inhibitory effect significantly varied with the duration (24-96 hours) of culture after transfection,with no interaction effect between the transfection with miR-145 mimics and culture duation (F =1.21,P =0.18).Compared with NC mimic-transfected cells,those transfected with miR-145 mimics showed a significant increase in the proportion of early apoptotic cells (18.9％ ± 4.1％ vs.4.3％ ± 1.2％,t =7.126,P ＜ 0.01),late apoptotic cells (9.3％ ± 2.3％ vs.3.6％ ± 1.6％,t =12.38,P ＜ 0.01),G1-phase cells (85.83％ ± 5.2％ vs.62.08％ ± 6.23％,t =11.78,P =0.007),but a significant decrease in the percentage of G2-phase cells (6.26％ ± 1.2％ vs.19.36％ ± 3.45％,t =7.610,P =0.017) and S-phase cells (7.91％ ± 1.3％ vs.18.56％ ± 5.23％,t =7.230,P=0.019).As luciferase assay showed,luciferase activity was significantly lower in HaCaT cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-wild carrying the wild-type 3'UTR of NRAS than in those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-wild (t =11.09,P =0.008),but similar between cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-mut carrying the mutant-type 3'UTR of NRAS and those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-mut (P ＞ 0.05).Real-time PCR and Western blot revealed that the overexpression of miR-145 mimics had no significant effect on NRAS mRNA expression (P ＞ 0.05),but significantly inhibited NRAS protein expression (1.52 ± 0.07 vs.0.20 ± 0.02,t =28.43,P＜ 0.01).Conclusion miR-145 might inhibit proliferation and promote apoptosis of HaCaT cells by influencing cell cycle via NRAS.

Objective To investigate the subtype distribution and sequence characteristics of HIV-1 strains prevalent among sexual infectors in Beijing. Methods We collected the blood samples from 100HIV sexual infectors in Beijing during 2008 and separated plasma specimens. RNA was extracted from the plasma and the gag gene was amplified by RT-PCR and nest-PCR. The PCR products were sequenced directly and phylogenetic analyses of gag gene was performed using the MEGA4 software. Results Among 100 HIV-1 plasma samples,84 gag gene fragments were amplified and analyzed. Eight HIV subtypes including B(22 strains), B'(8 strains),C( 1 strain) ,CRF01_AE (38 strains) ,CRF02_AG (2 strains) ,CRF07_BC(9 strains) ,CRF08_BC(3 strains) and C/CRF01_AE recombinant like strain( 1 strain) were identified circulating in Beijing. Conclusion CRF01 _AE and subtype B were predominant in Beijing account for 45.2% and 26.2% and the surveillance of HIV gene variation should be paid more attention.

OBJECTIVE: Structural equation modeling was used to explore the relationship between mental health and social psychological factors. METHODS: Using multi-stage sampling method,1 200 first-line production workers working in enterprises were chosen as study subjects. Chinese version of the WHO-5 well-being Index,and Social Psychological Factors Questionnaire(a self-designed questionnaire based on job demands control model) were used to investigate the mental health and social psychological factors. The structural equation modeling of social psychological factors on mental health was constructed. RESULTS: Among the 1 200 workers,about 34. 8% workers had poor psychological well-being(the score of the WHO-5 well-being Index is equal or less than 13). Job demands and job autonomy were available and have direct effect on work-related pressure [standardized path coefficient(β) were 0. 162 and-0. 186,P < 0. 05]. Job demands,job autonomy and work-related pressure were available have direct effect on mental health(β were-0. 136,0. 235 and-0. 135,P < 0. 05). Job demands can have an indirect effect on mental health through work-related pressure(β =-0. 022,P < 0. 05),and job autonomy can have an indirect effect on mental health through work-related pressure(β = 0. 025,P < 0. 05). CONCLUSION: The results of mental health and social psychological factors are consistent with the job demand control model. Job demands and work-related pressure are risk factors and job autonomy is a protective factor for mental health. Mental health intervention should be taken according to the related factors.

Cerebral blood flow directly affects substance metabolism and neural activity in the brain,and it is closely associated with the development and progression of neurodegenerative diseases.The ability for cerebral blood flow regulation directly affects the stability of cerebral blood perfusion,which is an important indicator for evaluating brain function.At present,studies have shown that the reduction in cortical perfusion is closely associated with cognitive decline in patients with Parkinson disease,and orthostatic hypotension often occurs in the late stage of the disease,thereby further increasing the risk of hypoperfusion,which reflects the importance of the ability for cerebral blood flow regulation in Parkinson disease.By reviewing related articles,this article summarizes the mechanism of cerebral blood flow regulation in patients with Parkinson disease,such as cerebral blood flow autoregulation,cerebral vasomotor reactivity,neurovascular coupling,and endothelium-dependent response,so as to provide new ideas and directions for exploring the effect of cerebral blood flow on Parkinson disease.

Objective: To analyze the effects of twirling reinforcing and reducing manipulations on protein expres-sion in parietal cortex of spontaneously hypertensive rats (SHRs), and elucidate the main mechanisms and differences between two manipulations in hypertension treatment. Methods: Rats were randomly divided into the control, model, twirling reinforcing manipulation (TRFM), and twirling reducing manipulation (TRDM) groups. The control and model groups received catch and fixation stimulations once a day for 14 days. The TRFM and TRDM groups were intervened once a day for 20 min for 14 days. On days 0, 2, 6, 10, and 14 after acupuncture, rat systolic blood pressures (SBPs) were measured. Differential protein (DP) expression in the rat parietal cortices was detected. Thereafter, GO functional significance and KEGG pathway enrichment analyses were performed. Results: Compared with the model group, SBP of rats in the TRDM and TRFM groups decreased on days 6 and 10 of acupuncture, respectively (P=.009; P<.001). Moreover, SBP of the TRDM group was signif-icantly lower than that of the TRFM group on days 10 and 14 of acupuncture (P = .015; P = .013). Compared with control group, 601 and 1040 DPs were up-and downregulated, respectively, in the model group. Compared with model group, 44 and 28 up-and downregulated DPs were expressed, respectively, in the TRFM group. Compared with model group, expression of 616 and 427 up-and downregulated DPs, respectively, was found in the TRDM group. After combining the results of GO and KEGG enrichment analyses, five and one pathways were found to be related to the central antihypertensive mechanism of the parietal cortex during twirling reducing and reinforcing manipulations, respectively. Conclusion: TRDM showed a more effective antihypertensive effect on SHRs than TRFM; this antihy-pertensive effect was related to the regulation of different proteins and their biological functions.

Objective To construct recombinant MVA vaccine encoding codon-optimized HIV-1 CRF01_AE gp145 gene and evaluate its immunogenicity in mice.Methods The intracellular region of codon-modified HIV-1 CRF01_AE consensus env gene obtained in our preliminary study was removed to get gp145 gene.The modified gp145 gene was cloned into shuttle plasmid pSC11.BHK-21 cells infected with wild-type MVA in advance were transfected with the recombinant shuttle plasmid.Then the recombinant MVA vaccine expressing gp145 rMVA-AEgp145 was obtained by homologous recombination.After expression of Gp145 was confirmed by WB assay,BALB/c mice were immunized with the recombinant MVA vaccine and the cellular immune responses were evaluated.Results Recombinant MVA vaccine expressed Gp145 protein efficiently and induced high level of cellular immune responses in immunized BALB/c mice.Conclusion The recombinant MVA vaccine encoding HIV-1 CRF01_AE gp145 gene was constructed successfully.This vaccine could induce strong env-specific CTL responses in mice.

OBJECTIVETo analyze the proportion and associated factors of taking subsequent confirmation test among men who have sex with men (MSM) after being tested positive in oral fluid HIV antibody test.

METHODSBy using successive sampling, 1 003 MSM, who were tested positive in oral fluid HIV antibody test in China-Bill & Melinda Gates Foundation AIDS prevention Program (Extension program) in Beijing during May 1 to December 31, 2013, were recruited. The inclusion criteria included: the objects were men who reported having sex with men; the objects aged more than 18 years old; the objects were tested positive in oral fluid HIV antibody test; the objects had not been reported as HIV positives in China Information System for Disease Control and Prevention previously. According to the program strategy, MSM grassroots organizations transferred the respondents to seek subsequent confirmation tests in specific Center for Disease Control and Prevention (CDCs) or hospitals. The subsequent confirmation tests included: fingertip blood HIV antibody rapid test, venous blood Enzyme Linked Immunosorbent Assay (ELISA) HIV antibody test and venous blood Western Blot (WB) HIV antibody test. Chi-square test was adopted to compare the proportion of taking subsequent confirmation tests in different groups. Nonconditional multivaritae binarylogistic regression analysis was taken to identify the associated factors with whether taking subsequent confirmation tests and to calculate the OR (95% CI) values.

RESULTSThe 1 003 respondents were (30.9 ± 9.1) years old. Among all objects, 87.8% (881/1 003) of them took fingertip blood HIV antibody rapid tests and the positive rate was 85.4% (752/881). 98.0% (737/752) of those who were identified as positive in fingertip blood HIV rapid tests took ELISA and WB tests, and the positive rate was 94.4% (696/737). Comparing with those who were expected to seek subsequent confirmation tests in CDCs, the OR (95% CI) value of those who were expected to seek tests in hospitals was 5.10 (1.69-15.36). The OR (95% CI) values of those who used condom sometimes and those who never used condom in anal sex were 5.81 (2.14-15.77) and 3.45 (2.00-5.97) respectively, in comparison with those who reported not having anal sex or using condom consistently in anal sex during the past 6 months. Comparing with the respondents recruited from the internet, the OR (95% CI) values of those recruited in bathrooms, parks/toilets and bars were 0.17 (0.05-0.53), 0.10 (0.04-0.29) and 0.22 (0.06-0.79) respectively. The likelihood of taking subsequent confirmation test decreased with the increase of number of male sexual partners in the past 3 months, and the OR (95% CI) value was 0.92 (0.86-0.99).

CONCLUSIONThe potential HIV positive MSM in the bathroom, park/toilet and bars are less likely to take subsequent confirmation test. Those who do not use condom consistently during anal sex are more likely to seek subsequent confirmation test. Medical organization conducting subsequent confirmation tests is more likely to increase the confirmation test rate of potential HIV positive MSM. The number of male sexual partners has negative correlation with whether to accept the subsequent confirmation test.

Beijing ; Condoms ; HIV Antibodies ; analysis ; HIV Seropositivity ; diagnosis ; Homosexuality, Male ; Humans ; Male ; Mass Screening ; Patient Acceptance of Health Care ; Risk-Taking ; Sexual Behavior ; Sexual Partners ; Surveys and Questionnaires

Objective To establish an ELISA for quantitative determination of decoy receptor 3 (DcR3) in human plasma.Methods A solid phase double antibody sandwich method was established for quantitative determination of DcR3.The anti-DcR3 antibody was immobilized onto ELISA plate.DcR3 in samples was captured by anti-DcR3 on ELISA plate and then detected by biotin-anti-DcR3 and subsequent peroxidase-labeled streptavidin,and the color was developed by adding substrate.The standard DcR3 samples on the same plate were detected simultaneously to calculate the DcR3 concentrations in unknown samples.The sensitivity,specificity,precision,recovery,linearity and DcR3 range in normal human adults were assessed.Results The sensitivity of the developed assay was 0.051 ng/mL.The intra-coefficient of variation (CV) was less than 10％ and inter-CV was less than 15％.The average recovery rate was 90.50％.When 2-fold amount of anti-TNF-α was added into the coated antibodies,10-fold amount of biotin-labeled anti-LIGHT,antiFAS or anti-TNF-α was added into the detection antibodies,or 10 fold amount of purified LIGHT protein was added into the standard DcR3 samples as competitor,no disturbing effects on standard curve were found.The linear range of the assay was from 0.25 to 16 ng/mL (r≥0.98).The concentration of DcR3 tested in 128 plasma samples from healthy adults was (0.21 ± 0.05) ng/mL with 95％ CI ranged from 0.14 to 0.28 ng/mL and no difference of age and sex was found.Conclusion The established ELiSA for determining plasma DcR3 exhibited high specificity,sensitivity,precision,fine linearity and wide detecting range.This method could be used for quantification of DcR3 in plasma.

OBJECTIVETo investigate the current supply and use of personal protective equipment (PPE) among rural-to-urban migrant workers in small and medium enterprises (SMEs) in Zhongshan and Shenzhen, China and the influential factors for the use of PPE, and to provide a basis for better occupational health services and ensuring the health of migrant workers.

METHODSMulti-stage sampling was used to select 856 migrant workers from 27 SMEs in Zhongshan and Shenzhen, and face-to-face questionnaire survey was conducted in these subjects. Statistical analysis was performed by one-way analysis of variance, chi-square test, and logistic regression.

RESULTSOf all migrant workers, 38.67%were supplied with free PPE by the factory, and this rate varied across industries (furniture industry: 45.81%; electronic industry: 31.46%) and SMEs (medium enterprises: 42.13%; small enterprises: 39.20%; micro enterprises: 22.16%); 22.43% insisted on the use of PPE. The logistic regression analysis showed that factors associated with the use of PPE included sex, age, awareness of occupational health knowledge, and the size of enterprise.

CONCLUSIONThe rates of supply and use of PPE among migrant workers are low. The larger the enterprise, the better the supply of PPE. Male gender, being elder, and high occupational health knowledge score were favorable factors for the use of PPE, while small enterprise size was the unfavorable factor for the use of PPE.

Adult ; China ; Female ; Humans ; Male ; Occupational Health Services ; statistics & numerical data ; Protective Devices ; statistics & numerical data ; Rural Population ; statistics & numerical data ; Transients and Migrants ; statistics & numerical data

Objective To determine the prognostic biomarkers and new therapeutic targets of the lung adenocarcinoma (LUAD), based on which to establish a prediction model for the survival of LUAD patients. Methods An integrative analysis was conducted on gene expression and clinicopathologic data of LUAD, which were obtained from the UCSC database. Subsequently, various methods, including screening of differentially expressed genes (DEGs), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and Gene Set Enrichment Analysis (GSEA), were employed to analyze the data. Cox regression and least absolute shrinkage and selection operator (LASSO) regression were used to establish an assessment model. Based on this model, we constructed a nomogram to predict the probable survival of LUAD patients at different time points (1-year, 2-year, 3-year, 5-year, and 10-year). Finally, we evaluated the predictive ability of our model using Kaplan-Meier survival curves, receiver operating characteristic (ROC) curves, and time-dependent ROC curves. The validation group further verified the prognostic value of the model. Results The different-grade pathological subtypes' DEGs were mainly enriched in biological processes such as metabolism of xenobiotics by cytochrome P450, natural killer cell-mediated cytotoxicity, antigen processing and presentation, and regulation of enzyme activity, which were closely related to tumor development. Through Cox regression and LASSO regression, we constructed a reliable prediction model consisting of a five-gene panel (MELTF, MAGEA1, FGF19, DKK4, C14ORF105). The model demonstrated excellent specificity and sensitivity in ROC curves, with an area under the curve (AUC) of 0.675. The time-dependent ROC analysis revealed AUC values of 0.893, 0.713, and 0.632 for 1-year, 3-year, and 5-year survival, respectively. The advantage of the model was also verified in the validation group. Additionally, we developed a nomogram that accurately predicted survival, as demonstrated by calibration curves and C-index. Conclusion We have developed a prognostic prediction model for LUAD consisting of five genes. This novel approach offers clinical practitioners a personalized tool for making informed decisions regarding the prognosis of their patients.