The effects of RNAi-mediated gene silencing of LRlG1 on proliferation and invasion of the human glioma cell line U251-MG and the possible mechanisms were explored in this study. The plasmids pGenesil2-LRIG1-shRNA1 and pGenesil2-LRIG1-shRNA2 were transfected into U251-MG glioma cells respectively by using Lipofectamine 2000 and the transfected cells in which the LRIG1 expression was stably suppressed were selected by G418. The cells transfected with negative shRNA served as control. The expression levels of LRIG1 mRNA and protein were measured by qRT-PCR and Western blotting, respectively. The cell cycle was analyzed by flow cytometry. The results showed that LRIG1 mRNA expression was reduced by 70% and 58% and LRIG1 protein expression by 58% and 26% in U251-MG cells transfected with pGenesil2-LRIG1-shRNAl and pGenesil2-LRIG1-shRNA2 relative to the negative shRNA-transfected U251-MG cells. The proliferative capacity of the LRIG1 specific siRNA-transfected cells was stronger than that of control cells. Cell cycle analysis showed that silencing LRIG1 significantly increased the percentage of S phase cells and the proliferation index (P<0.01). Moreover, silencing LRIG1 could promote the invasion of U251-MG cells (P<0.05). These findings suggested that LRIG1-targeting siRNA can exert a dramatically inhibitory effect on RNA transcription and protein expression of LRIG1, and LRIG1 down-regulation could promote the proliferation of U251-MG cells, arrest U251-MG cells in S phase, and enhance the invasion of U251-MG cells.

OBJECTIVETo study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.

METHODSPorcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).

RESULTSTen pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).

CONCLUSIONEffective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.

Androgens ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flavonoids ; Granulosa Cells ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; RNA, Messenger ; Swine

The identity of higher-order neurons and circuits playing an associative role to control renal function is notwell understood.We identified specific neural populations of rostral elements of brain regions that project multisynaptically to the kidneys in 3~ days after injecting a retrograde tracer pseudorabies virus (PRV)-614 into kidney of 13 adult male C57BL/6J strain mice.PRV-614 infected neurons were detected in a number of mesencephalic (e.g.central amygdala nucleus),telencephalic regions and motor cortex.These divisions included the preoptic area (POA),dorsomedial hypothalamus (DMH),lateral hypothalamus,arcuate nucleus (Arc),suprachiasmatic nucleus (SCN),periventricular hypothalamus (PeH),and rostral and caudal subdivision of the paraventricular nucleus of the hypothalamus (PVN).PRV-614/Tyrosine hydroxylase (TH) double-labeled cells were found within DMH,Arc,SCN,PeH,PVN,the anterodorsal and medial POA.A subset of neurons in PVN that participated in regulating sympathetic outflow to kidney was catecholaminergic or serotonergic.PRV-614 infected neurons within the PVN also contained arginine vasopressin or oxytocin.These data demonstrate the rostral elements of brain innervate the kidney by the neuroanatomical circuitry.

The distribution of shear stress on the bottom of the parallel plate flow chamber under different inlet velocities was analyzed by numerical simulation. In the present experimental study, the projection planes of the relative errors at 0.7% level were obtained, and then the efficient region and the actual entrance length were further corrected by introducing the concept of relative error. The results showed that the efficient region of the chamber increased with the direction of length while the inlet velocity was increased, and the actual entrance length was much greater than that of the theoretical entrance length. Therefore, in accordance to the needed range of shear stress in experiment and to the needed efficient region area, the optimum design of the flow chamber is necessary.
Algorithms ; Blood Flow Velocity ; Blood Pressure ; physiology ; Computer Simulation ; Humans ; Models, Cardiovascular ; Numerical Analysis, Computer-Assisted ; Pulsatile Flow ; Rheology ; Shear Strength ; Stress, Mechanical

OBJECTIVEThe purpose of this study was to explore the clinical significance of expression of Fas, CTLA-4 and RhoBTB2 genes in breast carcinoma.

METHODSSemi-quantitative reverse transcriptive polymerase chain reaction (RT-PCR) was used to analyze the mRNA expression levels of the three genes in tumor tissues from 60 patients with primary breast cancer and normal breast tissues of 30 cases. The relationship between gene expression and clinicopathological factors were analyzed and determined.

RESULTSThe relative expression levels (gray scale ratio between target gene and internal reference gene) of Fas, CTLA-4 and RhoBTB2 genes in breast carcinoma tissues were 0.699 +/- 0.285, 1.045 +/- 0.302 and 0.625 +/- 0.160, respectively. In the normal breast tissues, they were 0.502 +/- 0.178, 0.418 +/- 0.140 and 0.843 +/- 0.218, respectively. There were statistically significant differences of the expression of those three genes between carcinoma tissues and normal breast tissues (P < 0.01). The expression level of Fas in carcinoma tissues was significantly higher in lymph node matastasis positive patients (0.782 +/- 0.313) than that in node-negative patients (0.557 +/- 0.146, P < 0.01). The expression level of CTLA-4 gene in carcinoma tissues was lower in II stage patients (0.978 +/- 0.330) than that in III stage patients (1.134 +/- 0.240, P < 0.05). The expression level of RhoBTB2 gene was lower in invasive ductal carcinoma (0.597 +/- 0.157) than that in invasive lobular carcinoma (0.717 +/- 0.145, P < 0.05). There were no correlations of expression of the three genes at mRNA level and age, ER, PR, HER2 status and survival time. Furthermore, no correlation was seen among the three genes expression (P > 0.05).

CONCLUSIONThe expression of all the three genes at mRNA level is involved in genesis and progression of breast cancer. There exist correlations between Fas expression and axillary lymph node matastasis, CTLA-4 expression and disease stage, and RhoBTB2 expression and pathological type of breast cancer.

Adult ; Aged ; Antigens, CD ; genetics ; metabolism ; Breast ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; CTLA-4 Antigen ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Lobular ; metabolism ; pathology ; Female ; GTP-Binding Proteins ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Tumor Suppressor Proteins ; genetics ; metabolism ; fas Receptor ; genetics ; metabolism

Objective To explore risk factors of congenital malformations (CMs) and to evaluate its impacts on adverse pregnancy outcomes (APOs). Methods A prospective cohort study was conducted among pregnant women who received the first antenatal care from March 2013 to February 2016 in the reproductive center, obstetrics clinics, infertility clinics and ultrasound department of Hunan Provincial Maternal and Child Health Hospital. Corresponding information from pregnant women and their spouses were collected. Univariate and multivariate Logistic regression were used to screen possible risk factors of CMs and evaluate the impacts of CMs on other APOs. Results The study showed that women had history of non-standard BMI, smoking, hepatitis, pregnancy-related complications, gestational diabetes mellitus, infertility and using assisted reproductive technology before pregnancy; had no folic acid taking, active and passive smoking, drinking, uneven diet, high intensity physical activity during pregnancy increased the risk of CMs in offspring. Furthermore, the history of spouse smoking and eating betel nut also increased the risk of CMs in offspring. CMs might increase the risk of preterm birth, very preterm birth, low birth weight, very low birth weight, and perinatal mortality. Conclusions There are many risk factors of CMs. Knowing these risk factors, and giving them optimal prevention strategies and effective intervention measures are important measures in preventing the occurrence of CMs and other APOs.

Objective To investigate the surgical treatment of lung cancer invading the heart and large blood vessels.Methods The clinical data of 96 cases of lung cancer invading the heart and large blood vessel admitted into our hospital from March 2009 to March 2012 were retrospectively analyzed,especially the surgical methods,postoperative complications and 5-year survival rate.Results The 5-year survival rate of lung cancer patients who infringed the atrium and superior vena cava had no significant difference (P ＞ 0.05).However,the 5-year survival rate and median survival time of lung cancer patients who infringed the atrium and superior vena cava were significantly lower than those of lung cancer patients who infiltrated the pulmonary artery,and the difference was statistically significant(P ＜ 0.05).The median survival time of patients with lung cancer invading the atrium was lower than that of patients with lung cancer invading the superior vena cava,and the difference was statistically significant(P ＜ 0.05).The range of tumor involvement after operation was significantly lower than that before operation,and the difference was statistically significant(P ＜ 0.05).Three types of surgical methods were used for patients according to different invading sites,and the complication incidence of 3 surgical methods were similar with no statistically significant difference(P ＞ 0.05).Conclusion Surgical treatment for lung cancer invading the heart and large blood vessels has a good efficacy,which can improve the involved tumor range of patients without serious postoperative complications and provide opportunity for patients with comprehensive postoperative treatment.In addition,it can significantly improve the 5-year survival rate of patients.

The effects of RNAi-mediated gene silencing of LRIG1 on proliferation and invasion of the human glioma cell line U251-MG and the possible mechanisms were explored in this study.The plasmids pGenesi12-LRIG1-shRNA1 and pGenesi12-LRIGl-shRNA2 were transfected into U251-MG glioma cells respectively by using Lipofectamine 2000 and the transfected cells in which the LRIGI expression was stably suppressed were selected by G418.The cells transfected with negative shRNA served as control.The expression levels of LRIG1 mRNA and protein were measured by qRT-PCR and Western blotting,respectively.The cell cycle was analyzed by flow cytometry.The results showed that LRIG1 mRNA expression was reduced by 70％ and 58％ and LRIG1 protein expression by 58％ and 26％ in U251-MG cells transfected with pGenesil2-LRIGl-shRNAl and pGenesil2-LRIG1-shRNA2 relative to the negative shRNA-transfected U251-MG cells.The proliferative capacity of the LRIG1 specific siRNA-transfected cells was stronger than that of control cells.Cell cycle analysis showed that silencing LRIG 1 significantly increased the percentage of S phase cells and the proliferation index (P＜0.01).Moreover,silencing LRIG1 could promote the invasion of U251-MG cells (P＜0.05).These findings suggested that LRIG1-targeting siRNA can exert a dramatically inhibitory effect on RNA transcription and protein expression of LRIG1,and LRIG1down-regulation could promote the proliferation of U251-MG cells,arrest U251-MG cells in S phase,and enhance the invasion of U251-MG cells.

OBJECTIVETo study the association of single nucleotide polymorphisms (SNPs) of transcription factors (NKX2.5, GATA4, TBX5, and FOG2) with congenital heart disease (CHD) in the Chinese population.

METHODSPubMed, Google Scholar, CNKI, Wanfang Data, and Weipu Data were searched for articles on the association of SNPs of target genes with CHD in the Chinese population. If one locus was mentioned in at least two articles, the random or fixed effect model was used to perform a pooled analysis of study results and to calculate the pooled OR and its 95%CI. If a locus was mentioned in only one article, related data were extracted from this article to analyze the association between the SNPs of this locus and CHD.

RESULTSTwenty-three articles were included. The Meta analysis showed that there were significant differences between the CHD and control groups in the genotype and allele frequencies of GATA4 rs1139244 and rs867858 and the genotype frequency of GATA4 rs904018, while there were no significant differences in the SNPs of the other genetic loci between the two groups. The single-article analysis showed that there were significant differences between the two groups in the allele frequencies of NKX2.5 rs118026695/rs703752, GATA4 rs884662/rs12825/rs12458/rs3203358/rs4841588, and TBX5 rs6489956. There were no significant differences in the SNPs of FOG2 locus between the two groups.

CONCLUSIONSThe SNPs of some loci in NKX2.5, GATA4, and TBX5 are associated with CHD in the Chinese population, but the association between the SNPs of FOG2 locus and the development of CHD has not been found yet.

Asian Continental Ancestry Group ; genetics ; DNA-Binding Proteins ; genetics ; GATA4 Transcription Factor ; genetics ; Genetic Predisposition to Disease ; Heart Defects, Congenital ; genetics ; Homeobox Protein Nkx-2.5 ; genetics ; Humans ; Polymorphism, Single Nucleotide ; T-Box Domain Proteins ; genetics ; Transcription Factors ; genetics

OBJECTIVES: To investigate the incidence of preterm birth and risk factors for preterm birth. METHODS: A prospective cohort study was performed for the pregnant women in early pregnancy and their spouses, who underwent prenatal examination for the first time in Hunan Provincial Maternal and Child Health Care Hospital from May 2014 to December 2016 and decided to be hospitalized for delivery. A questionnaire survey was performed to collect exposure information possibly related to preterm birth. The hospital's medical record system was used for information verification and to record the pregnancy outcome. A multivariate logistic regression analysis was used to investigate the risk factors for preterm birth. RESULTS: A total of 6 764 pregnant women with complete data were included, and the incidence rate of preterm birth was 17.09%. The multivariate logistic regression analysis showed that a history of adverse pregnancy outcomes, eating areca nut before pregnancy, a history of pregnancy complications, a history of hepatitis, no folate supplementation during pregnancy, medication during pregnancy, active smoking and passive smoking during pregnancy, drinking during pregnancy, unbalanced diet during pregnancy, high-intensity physical activity during pregnancy, and natural conception after treatment of infertility or assisted conception as the way of conception were risk factors for preterm birth ( CONCLUSIONS There are many risk factors for preterm birth. Special attention should be paid to the life behaviors of pregnant women during pregnancy, and health education should be strengthened for pregnant women and their spouses to develop good living habits and reduce the incidence of preterm births.
Child ; Female ; Humans ; Infant, Newborn ; Pregnancy ; Pregnancy Outcome ; Premature Birth/etiology* ; Prospective Studies ; Risk Factors ; Tobacco Smoke Pollution