Oncogenic Gene CNOT7 Promotes Progression and Induces Poor Prognosis of Glioma.

Glioma is the most common malignant brain tumor in the central nervous system with the poor prognosis of patients. The CNOT7 (CCR4-NOT Transcription Complex Subunit 7) is an important functional subunit of CCR4-NOT protein complex that has not been reported in glioma. In this study, we aimed to explore the function of CNOT7 in glioma. The TCGA (The Cancer Genome Atlas) and CGGA (Chinese Glioma Genome Atlas) databases were used for investigating the expression and survival condition of CNOT7 in glioma. The cellular function experiments of qRT-PCR, CCK-8 assays, wound healing assays, and Transwell assays were conducted to verify the function of knockdown CNOT7 in the glioma cell lines DBTRG and U251. Enrichment analysis was used to explore the molecular mechanism of CONT7 in glioma. What is more, the upstream regulation transcription factors of CNOT7 were analyzed based on the ChIP-Atlas and cBioportal (provisional) databases, and verified by the qRT-PCR and luciferase reporter assay. The CNOT7 was highly expressed in glioma and presented the poorer prognosis. The knockdown of CNOT7 inhibited the proliferation, migration, and invasion of glioma cell line, compared to control group. The enrichment analysis revealed that the CNOT7 participated in the development of glioma via G2M checkpoint, E2F targets, IL6-JAK-STAT3, and TNF-α signaling pathways via NF-κB. Besides, it was found that the HDAC2 (Human histone deacetylase-2) contributes to increased CNOT7 expression in glioma. The high-expressed CNOT7 is an oncogene with poor prognosis and participate the progression of glioma.

Bromodomain containing 4 transcriptionally activated Deltex E3 ubiquitin ligase 2 contributes to glioma progression and predicts an unfavorable prognosis.

Background: Glioblastoma multiforme (GBM) is the most common type of glioma, and the most aggressive brain malignancy in adults. This study sought to identify novel survival-status related markers, and examine their function in glioma. Methods: The gene expression, survival heatmaps, and Kaplan-Meier survival plots of the genes were analyzed by using gene expression profiling interactive analysis (GEPIA) dataset, Linked Omics. The single-cell data analysis and tumor immune infiltration analysis was conducted by Tumor Immune Estimation Resource (TIMER) dataset. DBTRG and U251 cells with silenced Deltex E3 ubiquitin ligase 2 (DTX2) expression were constructed and used for Cell Counting Kit 8 (CCK-8), and wound healing assay in vitro. Chromatin immunoprecipitation sequencing (ChIP-seq) analysis was used to explore the histone activation marks and transcription factors DTX2 promoter. Dual-luciferase assays were carried out to detect the luciferase activities of bromodomain containing 4 (BRD4) binding to DTX2. Results: We first conducted a survival-status analysis to identify survival status-related genes in The Cancer Genome Atlas GBM and low-grade glioma data sets. A subsequent analysis identified 3 novel prognostic biomarkers; that is, DTX2, cytochrome P450 oxidoreductase, and Williams-Beuren syndrome chromosomal region 16 protein. In the validation Chinese Glioma Genome Atlas data sets, DTX2 showed the best performance, and was examined in a further analysis. Next, 3 short-hairpin ribonucleic acids were designed to silence DTX2 expression, and CCK-8 and wound-healing assays were applied to study the function of DTX2. We found that DTX2-silenced glioma cells exhibited a significant decrease in their growth and migration capabilities. Finally, the molecular basis for increased DTX2 in glioma was investigated via ChIP-Seq analysis and luciferase assays. The analysis revealed that DTX2 was transcriptionally activated by BRD4. Conclusions: In conclusion, BRD4 transcriptionally activates DTX2, contributes to glioma progression, predicts an unfavorable prognosis, and could provide new options for glioma prognosis prediction and treatment.

Hypomethylation-induced prognostic marker zinc finger DHHC-type palmitoyltransferase 12 contributes to glioblastoma progression.

Background: Glioma is the most common intracranial primary malignancy, characterized by abnormal signal transductions caused by transcriptional and post-transcriptional regulators. Studies show the palmitoylation of oncoproteins and tumor suppressors participate in cancer progression, while studies of protein S-palmitoyltransferases in glioma are limited. A systematic analysis of zinc finger DHHC-type palmitoyltransferases (ZDHHC) in glioma is still lacking. Methods: A prognostic heatmap and Kaplan-Meier overall survival plot of 24 members of the ZDHHC family in pan-cancer created. The expression and prognostic significance of ZDHHC12 was analyzed by using Gene Expression Profiling Interactive Analysis (GEPIA) and PrognoScan. DBTRG and U251 cells with silenced ZDHHC12 expression were constructed and used for cell counting kit-8 (CCK-8), Transwell assay and wound healing assay in vitro. Results: Here, we first conducted expression and prognostic analyses of 24 ZDHHCs from The Cancer Genome Atlas (TCGA), the Chinese Glioma Genome Atlas (CGGA), and other glioma datasets. We found ZDHHC12 to be the only unfavorable prognostic marker in glioma. The function of ZDHHC12 in glioma was then investigated with loss-of-function strategies and in vitro cell assays. Results showed that ZDHHC12 knockdown remarkably reduced the growth, migration, and invasion capabilities in DBTRG and U251 cell lines, suggesting that ZDHHC12 may contribute to malignant behavior in glioma cells. Finally, the molecular basis for ZDHHC12 expression in glioma was analyzed, and DNA hypomethylation was found to be responsible for increased ZDHHC12 mRNA expression and related prognoses. Conclusions: ZDHHC12 positively promoted the proliferation and migration of glioma cells. Decreased DNA methylation may lead to increased ZDHHC12 expression in gliomas. This study may deepen the understanding of glioma progression and therapeutics.

CRISPR screening of E3 ubiquitin ligases reveals Ring Finger Protein 185 as a novel tumor suppressor in glioblastoma repressed by promoter hypermethylation and miR-587.

Glioblastoma (GBM) is the most common and aggressive malignant primary brain tumor. E3 ligases play important functions in glioma pathogenesis. CRISPR system offers a powerful platform for genome manipulation, while the screen of E3 ligases in GBM still remains to be explored. Here, we first constructed an E3 ligase small guide RNA (sgRNAs) library for glioma cells growth screening. After four passages, 299 significantly enriched or lost genes (SELGs) were compared with the initial state. Then the clinical significance of SELGs were validated and analyzed with TCGA glioblastoma and CGGA datasets. As RNF185 showed lost signal, decreased expression and favorable prognostic significance, we chose RNF185 for functional analysis. In vitro overexpressed cellular phenotype showed that RNF185 was a tumor suppressor in two glioma cell lines. Finally, the molecular mechanism of decreased RNF185 expression was investigated and increased miR-587 expression and DNA hypermethylation was evaluated. This study would provide a link between the molecular basis and glioblastoma pathogenesis, and a novel perspective for glioblastoma treatment.

NPC2 as a Prognostic Biomarker for Glioblastoma Based on Integrated Bioinformatics Analysis and Cytological Experiments.

Glioblastoma (GBM) is one of the most common and fatal malignancies worldwide, while its prognostic biomarkers are still being explored. This study aims to identify potential genes with clinical and prognostic significance by integrating bioinformatics analysis and investigating their function in HNSCC. Based on the Single-cell RNA sequencing (scRNA-seq) results of H3K27M-glioma cells, computational bioinformatics methods were employed for selecting prognostic biomarker for GBM. The protein NPC2 (NPC Intracellular Cholesterol Transporter 2), which has been shown to be related to lipoprotein metabolism and innate immune system, was identified to be upregulated in GBM. NPC2 showed a relatively higher expression in GBM samples, and a negative correlation with tumor purity and tumor infiltrating immune cells. Additionally, NPC2 was knocked down in U87-MG and U251 cells line, and cell proliferation and migration capability were evaluated with CCK-8, scratch and transwell assay, respectively. Cytological experiments has shown that NPC2 overexpression inhibited GBM cells proliferation and migration, indicating its important role in GBM progression. This is the first investigation into the prognostic value of NPC2 interact with GBM. The potential molecular factor NPC2 have been identified as a prognostic biomarker for GBM.

Effect of improving the knowledge, attitude and practice of reproductive health among female migrant workers: a worksite-based intervention in Guangzhou, China.

UNLABELLED: Background The sexual and reproductive health (SRH) knowledge and attitudes of female migrant workers are far from optimum in China. A worksite-based intervention program on SRH-related knowledge, attitude and practice (SRH KAP) modification may be an effective approach to improve the SRH status among migrant workers. This study aimed to identify better intervention approaches via the implementation and evaluation of two intervention packages. METHODS: A worksite-based cluster-randomised intervention study was conducted from June to December 2008 in eight factories in Guangzhou, China. There were 1346 female migrant workers who participated in this study. Factories were randomly allocated to the standard package of interventions group (SPIG) or the intensive package of interventions group (IPIG). Questionnaires were administered to evaluate the effect of two interventions. RESULTS: SRH knowledge scores were higher at follow up than at baseline for all participants of the SPIG; the knowledge scores increased from 6.50 (standard deviation (s.d.) 3.673) to 8.69 (s.d. 4.085), and from 5.98 (s.d. 3.581) to 11.14 (s.d. 3.855) for IPIG; SRH attitude scores increased among unmarried women: the attitude scores changed from 4.25 (s.d. 1.577) to 4.46 (s.d. 1.455) for SPIG, and from 3.99 (s.d. 1.620) to 4.64 (s.d. 1.690) for IPIG; most SRH-related practice was also modified (P<0.05). In addition, after intervention, the IPIG had a higher knowledge level than the SPIG; the scores were 11.14 (s.d. 3.855) versus 8.69 (s.d. 4.085), and unmarried women in the IPIG had higher condom use rate than the SPIG (86.4% versus 57.1%). CONCLUSIONS: The interventions had positive influences on improvements in SRH knowledge, attitudes and behaviours. Additionally, IPIs were more effective than SPIs, indicating that a comprehensive intervention may achieve better results.

Promoting contraceptive use among female rural-to-urban migrants in Qingdao, China: a comparative impact study of worksite-based interventions.

BACKGROUND: We conducted a comparative study in worksites to assess the impact of sexual health promoting interventions on contraceptive use among female rural-to-urban migrants. STUDY DESIGN: In Qingdao ten manufacturing worksites were randomly allocated to a standard package of interventions (SPI) and an intensive package of interventions (IPI). The interventions ran from July 2008 to January 2009. Cross-sectional surveys at baseline and end line assessed the sexual behaviour of young female migrants. To evaluate the impact of the interventions we assessed pre- and post-time trends. RESULTS: From the SPI group 721 (baseline) and 615 (end line) respondents were considered. Out of the IPI group we included 684 and 603 migrants. Among childless migrants, self-reported contraceptive use increased significantly after SPI and IPI (adjusted odds ratio [aOR] = 3.23; 95% confidence interval [CI] = 1.52-6.84; p < 0.01 and aOR = 5.81; 95% CI = 2.63-12.80; p < 0.001, respectively). Childless migrants older than 22 years reported a greater use after IPI than after SPI. CONCLUSION: Implementing current Chinese sexual health promotion programmes at worksites is likely to have a positive impact on migrant women working in the manufacturing industry of Qingdao. More comprehensive interventions seem to have an added value if they are well targeted to specific groups.

Study on sexual and reproductive health behaviors of unmarried female migrants in China.

AIM: The purpose of this study was to broadly assess the level of knowledge, attitude and behaviors related to sexual and reproductive health (SRH) among unmarried female migrants in China. MATERIAL AND METHODS: This cross-sectional study was conducted and a self-administered questionnaire was designed for collecting information on SRH including 15 items for knowledge, 8 items for attitude and some items for contraception and abortion related behaviors. RESULTS: A total of 1690 unmarried female migrants were interviewed. Most of the respondents had less knowledge of SRH. Only one-third of respondents was aware of emergency contraceptives and could freely talk about SRH with their friends. Over one-third of respondents were not willing to come into contact with someone with AIDS or STDs. In this study, 10.4% participants had an unwanted pregnancy and 95% of them had an abortion. Multivariate analysis showed that having a boyfriend, duration of employment in city, knowledge on SRH and freely discussing SRH with peer were associated with having premarital sex among these unmarried female migrants. CONCLUSION: This study revealed that the unmarried female migrant was one of the most vulnerable groups concerning SRH. In some policy reforms, appropriate and cost-effective SRH services should be provided for these migrants.

Promoting contraceptive use more effectively among unmarried male migrants in construction sites in China: a pilot intervention trial.

Poor sexual and reproductive health status has been reported among rural-to-urban migrants in China. Therefore, some effective and feasible interventions are urgently needed. The authors developed a workplace-based intervention to compare 2 young labor migrant service packages (A and B) on the knowledge, attitude related to contraception, and contraceptive use among unmarried male migrants in Chengdu. Fourteen construction sites were randomly assigned to either of the 2 intervention packages. Interventions were completed in 3 months, and data were collected in 2 rounds independently (before and after interventions). After the intervention, the median scores for knowledge and attitude in migrants in package B were significantly higher than in migrants in package A. Although migrants in both packages increased use of condom, the increase was pronounced in migrants in package B, with odds ratio (OR) = 9.65 (95% confidence interval [CI] = 1.41-66.28). The rate of unwanted pregnancies was reduced more significantly in migrants in package B than in migrants in package A (OR = 0.16; 95%CI = 0.03-0.45). Unmarried male migrants who received the comprehensive intervention (package B) were more willing to use condoms and avoid unwanted pregnancies effectively.

Sexual and reproductive health status and related knowledge among female migrant workers in Guangzhou, China: a cross-sectional survey.

OBJECTIVE: The objective of this study was to investigate the current sexual and reproductive health (SRH) status including SRH-related knowledge and associated factors, self-reported symptoms of reproductive tract infection (RTI), medical assistance seeking behavior, sexual experience and contraceptive use, reproductive information approach and reproductive service utilization among female migrant workers in Huangpu district, Guangzhou city, China. STUDY DESIGN: A cross-sectional study was conducted in 2008 in eight factories, which were selected randomly from 32 eligible factories in the Huangpu district in Guangzhou. Descriptive statistics were used to describe the SRH status of migrant workers. Factors associated with the level of SRH knowledge were determined by a logistic regression model. RESULTS: Of 1346 female migrant workers, 831(61.7%) were unmarried and 515 (38.3%) were married. 27.2% of the unmarried respondents and 40.2% of the married respondents had suffered self-reported RTI symptoms. Among unmarried respondents, the median knowledge score was 5 points, compared to 8 points for the married. For unmarried migrant workers, factors associated with the knowledge level were age, education level, access to SRH information and service, sexual experiences and RTI symptoms. For married migrant workers, factors associated with the knowledge level were age, education level, access to SRH services and RTI symptoms. CONCLUSIONS: A high prevalence of self-reported RTI symptoms and a low knowledge level were found among young female migrant workers. Unmarried migrant workers are more vulnerable to SRH problems. Those findings demand more specific interventions targeting female migrants and in particular the unmarried.

Determinants of unmet need for contraception among Chinese migrants: a worksite-based survey.

BACKGROUND: Considerable sexual and reproductive health (SRH) challenges have been reported among rural-to-urban migrants in China. Predictors thereof are urgently needed to develop targeted interventions. STUDY DESIGN: A cross-sectional study assessed determinants of unmet need for contraception using semi-structured interviews in two cities in China: Guangzhou and Qingdao. RESULTS: Between July and September 2008, 4867 female rural-to-urban migrants aged 18-29 years participated in the study. Of these, 2264 were married or cohabiting. Among sexually-active women (n = 2513), unmet need for contraception was reported by 36.8% and 51.2% of respondents in Qingdao and Guangzhou, respectively; it was associated with being unmarried, having no children, less schooling, poor SRH knowledge, working in non-food industry, and not being covered by health insurance. A substantial proportion of unmarried migrants reported they had sexual intercourse (16.6 % in Qingdao and 21.4% in Guangzhou) contrary to current sexual standards in China. CONCLUSION: The study emphasises the importance of improving the response to the needs of rural-to-urban migrants and recommends strategies to address the unmet need for contraception. These should enhance open communication on sexuality, increase the availability of condoms, and improve health insurance coverage.

Mifepristone-induced early abortion and outcome of subsequent wanted pregnancy.

Follow-up information on subsequent pregnancies after mifepristone (RU486)-induced abortion is scarce. The authors examined whether one mifepristone-induced first-trimester abortion affects the outcome of a subsequent wanted pregnancy. In a study conducted in 1998-2001 at antenatal clinics in Beijing, Chengdu, and Shanghai, China, the authors enrolled 4,925 women with no history of induced abortion, 4,931 women with one previous mifepristone-induced abortion, and 4,800 women with one previous surgical abortion and followed them through pregnancy and childbirth. The adjusted odds ratio for preterm delivery in women with one mifepristone abortion compared with women with no abortion was 0.77 (95% confidence interval: 0.61, 0.98). Although the mean birth weight of infants born to women with mifepristone abortion was 33 g (95% confidence interval: 17, 49) higher than that of infants born to women with no abortion, the frequencies of low birth weight and mean lengths of pregnancy were similar. There were no significant differences in risk of preterm delivery, frequency of low birth weight, or mean infant birth weight in the comparisons of women with previous mifepristone abortion and women with surgical abortion. This study suggests that one early abortion induced by mifepristone in nulliparous women has no adverse effects on the outcome of a subsequent pregnancy.

[Study on the occurrence of cesarean section (CS) and factors related to CS in China].

OBJECTIVE: To find out the occurrence of cesarean section (CS) and to probe the factors associated with CS. METHODS: Women with CS as "case group" and women without CS as "control group" were chosen in a case-control study. RESULTS: Among 14 071 childbirth women, 6 421 had CS (case group) with the occurrence rate of 45.6% and 7 650 (54.4%) had normal delivery (control group). In comparison with the control group, the CS group had following several higher rates [with significant differences between case group and control group (P < 0.01)]: well-educated (78.9% vs 69.5%), white collar jobs (38.0% vs 32.3%), urban residents (79.1% vs 70.6%), high monthly income (>/= 500 Yuan) (81.0% vs 70.6%), of older age (>/= 25 years) (73.3% vs 63.0%), heavier baby weight (> 4 000 gram) (8.3% vs 2.9%), male babies (53.9% vs 51.4%), BMI of mother (> 24) (8.8% vs 4.8%), cephalopelvic disproportion (21.1% vs 0.9%), intrauterine asphysia (20.3% vs 6.7%), abnormality of force of labor (4.2% vs 2.7%), prolonged labor (2.9% vs 1.0%) and placenta previa (1.4% vs 0.4%). Our study also indicated that the higher the educational level was, the higher the rate of CS appeared; and the older the pregnant women was, the higher the rate of CS was. In CS group, over 70% primipara were over 24 years, and over 20% primipara had cephalopelvic disproportion and over 20% had intrauterine asphysia in CS group. CONCLUSIONS: At present, the occurrence rate of cesarean section was rather high (45.6%) in China. The high rate of CS was more likely to associate not only with abnormal physiological/medical factors (eg. cephalopelvic disproportion, intrauterine asphysia, abnormality of force of labor, and prolonged labour, etc.), but also with some demographic factors as education, occupation, income and age, etc. It is necessary to take measures to reduce the unnecessary CS in China.

[Association between low-dose oral contraceptive use and stroke in Chinese women].

OBJECTIVE: To investigate the association between low-dose combined oral contraceptive (COC) widely distributed in China and the risk of hemorrhagic stroke so as to decrease the adverse reaction of COC. METHODS: A prospective cohort study was made among 44 408 women who took hormonal contraceptives (HCs) and 75 230 women who used in trauterine device (IUD) in 25 townships and toens in Rudong County and Taicang City, Jiangsu Province, to compare the incidence of stroke among them from 1 July 1997 to 30 June 2000. RESULTS: (1) The incidence of hemorrhagic stroke (age-and-county standardized rates) was 34.74 per 100 000 py in the HC cohort, 2.72 times higher than that in the IUD cohort (P < 0.01). The incidence of hemorrhagic stroke among women less than 45 years of age was remarkably higher in HC cohort than in the IUD cohort. The relative risk of incidence of hemorrhagic stroke was 2.17 (1.16, 4.06) among the HC users who had stopped HC use for more than 10 years, still significantly higher than among the IUD users. (2) The relative risk of incidence of hemorrhagic stroke among the women who used low-dose COCs during the survey was 3.60 (1.73, 7.53) times higher than that among the IUD users. The relative risk of incidence of hemorrhagic stroke was 3.09 (1.26, 7.57) among the women who had stopped low-dose COC use for less than five years, however, the risk of hemorrhagic stroke remarkably decreased after they stopped taking COC for more than 5 years. (3) No obvious increase in incidence of infarctional stroke was found among the women who took low-dose COC during the survey. (4) Hypertension might be the most important risk factor of hemorrhagic stroke among the low-dose COC users. CONCLUSION: There is an obvious increase in risk of hemorrhagic stroke among the Chines women who take low-dose of China-made COCs for a long time. The efffect of China-made COCs on pathogenesis of hemorrhagic stroke lasts sometime after the discontinuation of COC use.