The relevance of EZH2 polymorphism to breast cancer risk in Chinese females: results from a multicenter case-control study / 中华普通外科杂志

Objective:To investigate the relationship between single nucleotide polymorphism (SNPs) of Zeste homolog enhancer 2 (EZH2) gene and the risk of breast cancer.Methods:Recruiting 1 039 breast cancer patients and 1 040 controls at 22 referral hospitals nationwide in China, the genotype distribution of 3 SNPs loci of EZH2 genes was observed to detect the correlation between different genotypes and the risk of breast cancer genotypes EZH2 expression in breast cancer tissues and its correlation with patient prognosis were analyzed using breast cancer data from the database.Results:EZH2 rs6464926 CC genotype was compared with TT genotype (TT vs. CC: OR=1.362, 95% CI: 1.063-1.746, P=0.015) and dominant model (TC+TT vs .CC: OR=1.22, 95% CI: 1.004-1.483, P=0.045) .In women with BMI ≥24 kg/m 2, the TC genotype ( P=0.050), TT genotype ( P=0.025) and dominant model (TC+TT, P=0.021) of rs6464926 locus were significantly different from CC genotype in cancer risk. rs6464926 was correlated with EZH2 gene expression ( P=6.89E-47). EZH2 gene is highly expressed in breast cancer tissues, and patients with high expression were associated with shorter OS ( HR=1.27, P=0.013), DMFS ( HR=1.37, P<0.01), and RFS ( HR=1.44, P<0.01). Conclusions:The polymorphism rs6464926 of EZH2 gene is associated with breast cancer susceptibility in Chinese women. rs6464926 might regulate breast cancer risk and prognosis by changing EZH2 expression.

Improve diagnostic performance of PCOS by measurements of testosterone and dehydcoepiandrosterone sulfate with ultra-performance liquid chromatography tandem mass spectrometry / 中华内分泌代谢杂志

Objective To compare ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) and chemiluminescence immunoassay(CLIA) measurement of human serum androgen levels in polycystic ovarian syndrome(PCOS). Methods 160 PCOS patients and 146 healthy subjects(control group) were recruited and their serum androgen levels——measurements of testosterone and dehydcoepiandrosterone sulfate (DHEA-S) were tested by UPLC-MS/MS and CLIA. The androgen results combined with body mass index(BMI) were analyzed by ROC curve. According to area under curve(AUC) calculated by SPSS, a better method will be selected to provide accurate test results for physicians. Results (1)AUC of DHEA-S tested by UPLC-MS/MS was significantly higher than the one tested by CLIA(P<0.01). There was no significant difference between the AUC of T tested by UPLC-MS/MS and the one tested by CLIA. (2)AUC of T combined with DHEA-S tested by HPLC was not only significantly higher than the AUC of two combined indicators tested by CLIA(P<0.01),but also significantly higher than the AUC of a single indicator——either T(P<0.01) or DHEA-S(P<0.01) tested by UPLC-MS/MS. (3)The AUC of T,DHEA-S combined with BMI tested by HPLC was not only significantly higher than the AUC of three combined indicators tested by CLIA(P<0.01),but also higher than the AUC of two combined indicators tested by UPLC-MS/MS(P<0.05). Conclusion T and DHEA-S tested by UPLC-MS/MS combined with BMI has a certain reference value in the diagnosis of PCOS.

Study of The Effect on Islets Cultured In vitro and STZ-induced Diabetic Rats In vivo of EGF-like Domain of Betacellulin / 生物化学与生物物理进展

Betacellulin (BTC) is one of "islets regeneration factors" which received more and more attention these years. BTCe is the C-terminal 50-residue region of mature BTC protein and can bind with erbB-1?erbB-4 receptor. It has the same mitogenic activity on cells as the whole section. BTC and BTCe can improve the level of glucose-stimulated insulin secretion (GSIS) during islets culture in vitro though they had no effect on the acute insulin secretion. BTCe also effectively ameliorated the hyperglycemia of STZ-induced diabetic rats by a single plasmid injection into muscle of rats. It is supposed that BTCe promote the proliferation of PDX-1 positive cells or repair some signal transduction pathway. Perhaps the latter is more important.

Identification of variations of gene expression of visceral adipose and renal tissue in type 2 diabetic rats using cDNA representational difference analysis / 中华医学杂志(英文版)

<p><b>OBJECTIVES</b>To identify differences in gene expression in renal and visceral adipose tissue in type 2 diabetic rats using cDNA representational difference analysis (RDA) and to explore the molecular pathogenesis of type 2 diabetes and its chronic vascular complications.</p><p><b>METHODS</b>A rat model of type 2 diabetes was generated by administration of a high fat and calorie diet combined with a low dose of streptozocin (STZ) injected into the tail vein. The difference bands were generated by cDNA representational difference analysis (cDNA RDA). The final difference products were ligated into the pUC-18 vector and sequenced. A bioformatics analysis was performed on the obtained expressed sequence tags (ESTs), and then the expression levels of known and novel genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). At the same time, full-length cDNA of a novel gene was cloned in silico.</p><p><b>RESULTS</b>The type 2 diabetic rats in this experiment experienced hyperglycemia, lipidemia, lower insulin sensitivity and normal body weight. We obtained 9 novel ESTs and 2 novel genes from renal tissue of rats and 6 novel ESTs and 1 known gene, the rat lipoprotein lipase (LPL) gene from their visceral adipose tissue. The 2 novel genes (RS91 and RS2) from the renal tissue were both very similar to serine (or cysteine) proteinase inhibitor, clade F and eukaryotic translation initiation factor 3 and subunit 5 (EIF-3 epsilon). The expression of both novel genes and the LPL gene were upregulated in renal and visceral adipose tissue of type 2 diabetic and fat-enriched rats. Full-length cDNA of the novel gene RS91 was cloned in silico.</p><p><b>CONCLUSIONS</b>(1) The rat model of type 2 diabetes generated in this study was ideal because the disease in the animals closely mimicked type 2 diabetic patients. (2) cDNA RDA is a flexible, inexpensive, more accurate, sensitive and highly effective technique for identifying differences in gene expression. (3) Six novel ESTs and 1 known gene were obtained from rat visceral adipose tissue. The LPL gene was upregulated in adipose tissue of type 2 diabetic and fat-enriched rats, a result possibly related to the diabetic animals' high fat and calorie diet, lipidemia, insulin resistance (RI) and molecular pathogenesis. (4) Nine novel ESTs and 2 novel genes were obtained from the renal tissue of rat. We believe the 2 novel genes to be the serine proteinase inhibitors clade F and EIF-3 epsilon in rats. The upregulation of the 2 novel genes in renal tissue of type 2 diabetic rats and may have been related to their renal impairment.</p>

Expression and identification of type 1 diabetes associated autoantigen IA-2 / 中华医学杂志(英文版)

<p><b>OBJECTIVES</b>To obtain prokaryotic expressed IA-2 recombinant protein and to identify its immunological activity.</p><p><b>METHODS</b>The complimentary DNA (cDNA) coding for the intracytoplasmic part of IA-2 (IA-2ic) was amplified from human fetal brain RNA, and was subcloned into the PinPoint Xa-1 T vector to construct recombinant expression plasmid, and was then expressed in E. coli JM109 cells as a fusion protein with a biotinylated peptide sequence at the aminoterminus. The biotinylated fusion protein was then purified by affinity chromatography and was subsequently dialyzed. Finally, its immunogenicity was evaluated by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The purified IA-2ic fusion protein resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a single Coomassie brilliant blue stained band with a molecular weight of 59 kDa and its immunogenicity was confirmed by ELISA.</p><p><b>CONCLUSIONS</b>E. coli expressed IA-2ic fusion protein has immunological activity. It can be used for detection of IA-2 autoantibodies (IA-2A) and for further studies on type 1 diabetes in future.</p>

Identification of difference of gene expression in the visceral adipose tissue of the lean type 2 diabetic rats / 中华内分泌代谢杂志

Objective To investigate the difference of gene expression in the visceral adipose tissue of the lean type 2 diabetic rats. Methods The difference bands were generated by the representational difference analysis ( cDNA RDA) ; the final difference products was ligated into the pUC-18 vector, subcloned and sequenced, and the obtained expressed sequence tags ( ESTs) were given bioinformatics analysis, and then the expression level of known gene was verified by semi-quantitative RT-PCR preliminarily. Results Six novel ESTs and 1 known gene [ rat lipoprotein lipase ( LPL) gene) were obtained in visceral adipose tissue of rat. One of the ESTs was partially similar to mouse musculus ERA-like GTPase (Era) gene. LPL gene was up-regulated in the visceral adipose tissue of the lean type 2 diabetic rats and those fed with fat-enriched food. Conclusion Six novel ESTs and 1 known gene in high expression are obtained from visceral adipose tissue of rat. LPL gene among them is up-regulated and may be related to the high fat and high calorie diet, hyperlipidemia, insulin resistance and molecular pathogenesis of the lean type 2 diabetic rat.

Electronic cloning and functional prediction of new apoptosis related gene 2ass-bnip3 of diabetic cardiomyopathy / 中华内分泌代谢杂志

Using the technique of fluorescent-labled mRNA differential display, new apoptosis related gene 2ass-bnip3 of type 2 diabetic cardiomyopathy was found, the sequence of 1594 bp with coding 187 amino acids was obtained by the full-length clone, and its structural and functional predictions were performed. 2ass-bnip3 may play an important role in the development of diabetic cardiomyopathy via a regulatory pathway of calcium regulation and apoptosis.