Objective To access the role of methylated RUNX3 ggene in the carcinogenesis an progression of ovarian carcinoma.Methods Sample of 32 epithelial ovarian carcinoma tissues,32 para-carcinoma tissues,36 benign epithelial tumors and 10 normal ovarian tissues and 2 cell lines(3A0 and SKOV3),were collected and subject to methylation-specific PCR(MSP).Promoter methylation status of RUNX3 in two tumor cell lines were analyzed before and after 5-aza-2'deoxycytidine treatment.In addition. mRNA expression of RUNX3 were investigated by quantitative reverse transcription-PCR Results CpG island methylation of RUNX3 was observed in 53.1%(17 of 32)of epithelial ovarian cancer,and 37.5% (12 of 32)in corresponding noncancerous tissues,16.7% in benign epithelial tumors(6 of 36),and all celllines,but not in normal control tissues.The prevalence of RUNX3 gene CpG methylation in malignant was significantly higher than those in benign and normal tissues(X2=10.060,X2=8.925,P<0.05). Nineteen percent(6 of 32)of ovarian epithelial carcinoma expressed RUNX3 mRNA,while its expression Was present in 28% (9 of 32)corresponding noncancerous tissues and 72% (26 of 36)of benign ovarian tumor and 80% (8 of 10)of normal ovarian tissues.The RUNX3 promoter methylation was found in all cell lines tested.The ratio of expression of RUNX3 mRNA in ovarian epithelial carcinoma was significantly lower than those of normal and benign tumors(X2=19.443,X2=12.862,P<0.05).After 5-aza-2'deoxycytidine treatment,methylation was partially or completely reversed,and its mRNA expression was increased.The relationship between gene expression and promoter methylation was reversely correlated.Conclusions Our results suggest that promoter hypermethylation of RUNX3 genes is common in ovarian cancer. Therefore, hypermethylation of RUNX3 genes may be involved in the carcinogenesis of ovarian cancer and may serve as an early diagnostic marker for ovarian cancer. The close correlation between RUNX3 methylation of its mRNA suggests that methylation which can be reversed.Thus,it provides a new way for therapy of ovarian Cancer.

Objective To observe the effects of 5-Aza-2’-deoxycytidine(5-Aza-CdR)on cell proliferation and apoptosis in ovarian cancer cell line SKOV3 and the expression of mismatch repair gene HMLH1/HMSH2, and to investigate the potential mechanism of its antitumorigenesis. Methods Human ovarian cancer cell line SKOV3 was treated with 5-Aza-CdR(0.5, 5 and 50 ?mol/L), a specific demethylation agent for 3 d, and then cultured in RPMI-1640 medium for 7 d.The growth of cells was examined by MTT assay. The apoptosis was analyzed by flow cytometry. The expression of HMLH1 and HMSH2 mRNA was observed by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). Results SKOV3 cells treated with 5-Aza-CdR displayed a slow growth rate in comparison with that of the control cells, The apoptosis rates of each group were 10.59%?1.57%、17.52%?1.72%、34.10%?1.45%,respectively,which were markedly higher than that of control(P

Objective To investigate the association between bone mineral density (BMD) and left ventricular mass index (LVMI) in elderly men in Beijing.Methods Totally 370 elderly men with an average age of (76.6±9.3) years from the departments of gerontology were included.BMD,echocardiography measurements as well as blood chemistry were analyzed.LVMI was obtained by echocardiography.All the subjects were divided into two groups:non-LVH group (n=231) and LVH group (n =139).Differences in quantitative variables were tested by independent-sample t test.Multiple stepwise linear regression analysis were performed to identify determinants of LVMI.Results The serum creatinine concentration was significantly higher in LVH group than in non-LVH group [(97.1±43.0) μmol/L,(88.2±21.1) μmol/L (P＜0.05)].Compared with non-LVH group,LVH group showed that the lumbar spine BMD (L1-L4) were significantly lower[L1:(0.90±0.16) g/cm2 vs.(0.95±0.21) g/cm2,P=0.05; L2:(0.95±0.17) g/cm2 vs.(1.01±0.20) g/cm2,P＜0.01 ; L3:(0.99±0.19) g/cm2 vs.(1.06±0.28) g/cm2,P＜0.01] as well as the lumbar spine totalBMD [(0.97±0.18) g/cm2 vs.(1.03-1-0.26) g/cm2,P＜0.05].The femur BMD was lower in theLVH group than in non-LVH group [trochiter:(0.64±0.11) g/cm2 vs.(0.67±0.17) g/cm2,P＜0.05; inter area:(1.00±0.17) g/cm2 vs.(1.05±0.22) g/cm2,P＜0.05].Multiple stepwise linear regression analysis demonstrated that BMI (r=0.27,P＜0.01),the lumbar spine BMD (r=-0.20,P＜0.01),age (r=0.16,P＜0.05),serum creatinine (r=0.15,P＜0.05) were independently correlated with LVMI.Conclusions In elderly men in Beijing,the lumbar spine BMD is an independent correlative factor for LVMI.

Objective To investigate the effect of 5-aza-2'deoxycytidine (5-Aza-CdR) on proliferation and expression of RASSF1 A gene in human ovarian cancer cell line SKOV3 and 3AO.Methods SKOV3 and 3AO cells were treated with different concentrations (0.5,5,50 μmol/L) of DNA methyltransferase inhibitor 5-Aza-CdR.RT-PCR and Western Blot were adopted to detect expression of mRNA and protein of RASSF1A gene before and after treatment with 5-Aza-CdR respectively.Results Compared with control group,when the 5-Aza-CdR concentration was 0.5,5,50 μmol/L after drug treatment,human ovarian cancer cells could significantly inhibit tumor cell growth; SKOV3 and 3AO cells in control group were observed weaker expression of RASSF1A mRNA.After treated with 5-Aza-CdR,the expressions of RASSF1A mRNA were observed increased with the increase of the drug concentration.After treated with different concentration of 5-Aza-CdR,the expressions of RASSF1A mRNA treated with 0.5 μmol/L 5-Aza-cdR was lower than those treated with 5 and 50 μmol/L 5-Aza-cdR (t =-8.866,P =0.01 ; t =-12.256,P =0.000).However,expressions of RASSF1A mRNA treated with 5 and 50 μmol/L 5-Aza-cdR respectively showed no statistical significance (t =0.431,P =0.689).Expressions of RASSF1A protein treated with 0.5 μmol/L 5-Aza-cdR and 5 μmol/L 5-Aza-cdR didn't show statistically significant (t =-1.586,P =0.188).Conclusion Expressions of RASSF1A mRNA and protein in SKOV3 and 3AO cells were evidently enhanced.As one kind of methyltransferases inhibitors,5-Aza-CdR can inhibit ovarian cancer cell line SKOV3,3AO growth through the RASSF1A promoter methylation,and thus promote their apoptosis.

Objective To investigate the effects of Matrine on proliferation and apoptosis of human cervical carcinoma cell line Hela in vitro. Methods Human cervical carcinoma cell line Hela was cultured in vitro. Cell proliferation was assessed by MTT colorimetric assay. The percent age of apoptosis cells was detected by TUNEL staining. The expression of PCNA protein was displayed with immunohistochemistry. Results Proliferation of Hela cells was inhibited in 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0g/L Matrine-treated groups compared with those in the control group(P

ObjectiveTo investigate the change of gene polymorphorism of surfactant protein-B (SP-B) intron 4 in infants with bronchopulmonary dysplasia (BPD).MethodsForty-five infants with BPD (BPD group) and ninety-nine infants without lung diseases (control group) who admitted into Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology from July 2008 to July 2011 were selected into this study.Genotyping for fragment length polymorphism of SP-B intron 4 was performed by polymerase chain reaction (PCR),agarose gel electrophoresis,cloning and sequencing methods in both groups.Differences of allele frequencies (invariant allele and variant allele) and genotype frequencies (invariant genotype and variant genotype) between BPD group and control group were analyzed.The differences of gestational age and birth weight between the two groups were compared with Independent-Samples t test.The gender composition and differences of allele or genotype frequencies between the two groups were compared with Chi-square test.Results Invariant allele frequencies in BPD group and control group were 83.3％ (75/90) and 92.0％ (182/198),and variant allele frequencies were 16.7％ (15/90,including eight insertion alleles and seven deletion alleles) and 8.1％ (16/198,including eight insertion alleles and eight deletion alleles).There were significant differences between the two groups (x2 =4.75,P =0.029).In BPD group,there were 32 cases (71.1 ％,32/45) invariant genotypes and 13 cases (28.9 ％,13/45,including seven cases insertions and six cases deletions) variant genotypes; in the control group,there were 85 cases invariant genotypes (85.8％,85/99) and 14 cases (14.1％,14/99,six insertions and eight deletions) variant genotypes.Significant difference was found between the two groups (x2=4.42,P＜0.036). ConclusionsVariations of SP-B intron 4 were more in BPD infants,and the variation of SP-B intron 4 might be associated with BPD.

Objective To explore the relationship between serum 25-hydroxy vitamin D level and prostatic hyperplasia in elderly men.Methods Totally 95 male patients aged over 60 years were included.Blood pressure,body weight,body height,body mass index (BMI) were measured and calculated.Venous blood samples were obtained to determine fasting serum levels of 25-hydroxy vitamin D3 and blood glucose (FBG),total cholesterol (TC),triglycerides (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),calcium and prostate specific antigen (PSA),total prostate volume(PV) and annual prostate growth rate were measured and calculated by ultrasound.Results The serum 25 (OH) D3 levels were varied from 12.1 nmol/L to 83.9 nmol/L,with an average of (35.5±15.2) nmol/L in elderly male patients.PV growth rate were significantly lower in elderly men with 25 (OH) D3 ＞ 50 nmol/L than in elderly men with 25 (OH) D3≤50nmol/L[(31.5± 6.0) mlvs.(39.9 ± 14.5) ml,(0.4± 0.2) ml/yvs.(0.5 ± 0.4) ml/y,P＜0.001 or 0.01].PV was negatively correlated with serum 25-hydroxy vitamin D3 level (r=-0.207,P＜0.05),and positively correlated with BMI and PSA (r=0.297,0.958,P＜0.05 or and P＜0.001).While annual prostate growth rate was positively correlated with BMI and PSA (r=0.316,0.464,P＜0.01 or ＜0.001),and positively correlated with serum 25-hydroxy vitamin D3 level,but the difference was not statistically significant (P＞0.05).Conclusions Low serum 25-hydroxy vitamin D3 level may play a role in the pathogenesis of prostatic hyperplasia.

BACKGROUND: As a primary clinical predictor of fracture risk, bone mineral density (BMD) is partly genetically determined. Osteoprotegerin (OPG) is one important candidate gene in the pathogenesis of osteoporosis. OBJECTIVE: To investigate the association between T245G polymorphisms in the OPG gene and BMD. METHODS: A total of 281 elderly men and postmenopausal women, 182 males and 99 females, who received routine examinations at Peking University People's Hospital between September 2008 and April 2010 were included in this study. T245G polymorphisms in the OPG gene was detected by polymerase chain reaction-restriction fragment length polymorphism together with DNA sequencing. The BMD of the lumbar spine, Ward's triangle, and forearrm was determined by dual energy X-ray absorptiometry. Clinical variables and biochemical measurements were collected simultaneously. The association between T245G polymorphisms and each detection index was analyzed using analysis of variance. RESULTS AND CONCLUSION: The distribution of T245G genotype (alleles T, G) had no difference in elderly men or postmenopausal women (P > 0.05). The GG genotype and TG genotype had higher lumbar spine BMD and TT genotype had lower lumbar spine BMD (P < 0.05). There was no difference in BMD of the Ward's triangle or forearm among different genotypes (P > 0.05). Association between T245G polymorphism and BMD was not found in postmenopausal women. These findings indicate that OPG gene is related to lumbar spine BMD in elderly men.

Objective To evaluate the relationship between insulin resistance and benign prostatic hyperplasia (BPH).Methods Totally 150 male patient from the Department of Geriatrics in Peking University Hospital were included in this study.Blood pressure,body weight,body height,body mass index (BMI) were measured and calculated.Biochemical analyses including serum fasting levels of insulin(FINS),glucose,total cholesterol,triglycerides,low-density lipoprotein cholesterol,high density lipoprotein cholesterol,and prostate-specific antigen (PSA) were performed.Total prostate volume (PV) were measured by ultrasound.Results PV and annual prostate growth rate were more increased in insulin resistance group(40 cases) compared with insulin sensitivity group(110 cases) (t=2.91,3.71 respectively,both P＜0.01).Along with the levels of FINS,HOMA-IR and PSA were increased,the prostate volume was enhanced (t=-3.02,-2.88,-2.84 respectively;all P ＜0.05).PV was positively correlated with insulin resistance,serum fasting insulin and PSA (r=0.16,0.16,0.35;all P＜0.05),while annual prostate growth rate was positively related with insulin resistance,serum fasting insulin,PSA and BMI (r =0.22,0.21,0.24,0.19 ; all P ＜ 0.05).Conclusions Insulin resistance and fasting insulin plays roles in the pathogenesis of prostatic hyperplasia.

Objective:To analyze the clinical characteristics and SLC25A13 gene mutation in children with neonatal intrahepatic cholestasis caused by citrin deficiency(NICCD).Methods:The data of 18 children diagnosed with NICCD in Xi′an Children′s Hospital from January 2014 to December 2018 were collected.The clinical manifestations, biochemical characteristics, SLC25A13 gene mutation and prognosis were analyzed.Results:All the 18 cases of NICCD were from North China and the age of initial diagnosis averaged(63.4±19.5)days.The clinical manifestations included jaundice(100%), light yellow or white stool(38.9%), growth retardation(27.8%)and so on.All patients had cholestasis.Of 18 cases, the levels of glutamyltranspeptidase, total bile acid and alpha fetoprotein were all increased, and serum albumin was decreased.Elevated aspartate aminotransferase(94.4%), elevated glutamic pyruvic transaminase(72.2%), prolonged prothrombin time(88.9%), hyperlactemia(83.3%), hypoglycemia(77.8%), anemia(66.7%)and other biochemical abnormalities were observed.Citrulline and other serum amino acids of all cases were elevated in blood samples by tandem mass spectrometry.The increase of 4-hydroxyphenyllactate and 4-hydroxyphenylpyruvate was found in 70%(7/10)urine samples by gas chromatography.Age was negatively correlated with total bile acid( r=-0.469, P=0.049), and positively correlated with blood ammonia, threonine, methionine, ornithine and tyrosine( r=0.472, 0.690, 0.698, 0.678 and 0.769, respectively, P<0.05). A total of 16 SLC25A13 gene mutations were detected, of them c. 851_854del(33.3%)and c. 1638_1660dup(19.4%)were the most common.c.1841+ 3_1841+ 4del, c.980_981del(p.E327Vfs*45)and c. 602A>T(p.E201V)were novel mutations.Among the 17 children who were followed up, 1 case died and 16 cases had normal biochemical parameters within 1 year. Conclusion:The characteristic biochemical changes are helpful for early recognition of NICCD.The prognosis of NICCD is good if the treatment is appropriate and timely.c.851_854del and c. 1638_1660dup are high-frequency mutations of SLC25A13 gene in north China.