Toll-like receptor 9 polymorphisms and Helicobacter pylori influence gene expression and risk of gastric carcinogenesis in the Brazilian population

BACKGROUND Toll-like receptors(TLRs)are the first line of host defense,and are involved in Helicobacter pylori(H.pylori)recognition and activation of both inflammatory and carcinogenic processes.The presence of single nucleotide polymorphisms(SNPs)in genes that activate the immune response may modulate the risk of precancerous lesions and gastric cancer(GC).Among them,Toll-like receptor 9(TLR9)polymorphisms have emerged with a risk factor of infectious diseases and cancer,however the studies are still inconclusive.AIM To evaluate whether TLR9 rs5743836 and rs187084 SNPs contribute to the risk of gastric carcinogenesis,and its influence on mRNA expression.METHODS A case-control study was conducted to evaluate two TLR9 SNPs(TLR9-1237 TCrs5743836 and TLR9-1486 CT-rs187084)in chronic gastritis(CG)and GC patients.A total of 609 DNA samples of peripheral blood[248 CG,161 GC,and 200 samples from healthy individuals(C)]were genotyped by polymerase chain reaction-restriction fragment length polymorphism.All samples were tested for the H.pylori infection using Hpx1 and Hpx2 primers.Quantitative polymerase chain reaction by TaqMan?assay was used to quantify TLR9 mRNA from fresh gastric tissues(48 GC,26 CG,and 14 C).RESULTS For TLR9-1237,the TC+CC or CC genotypes were associated with a higher risk of GC than C[recessive model odds ratio(OR)=5.01,95%confidence interval(CI):2.52-9.94,P<0.0001],and the CG(recessive model OR=4.63;95%CI:2.44-8.79,P<0.0001)groups.For TLR9-1486,an association between the CT+TT genotypes and increased risk of both GC(dominant model OR=2.72,95%CI:1.57-4.72,P<0.0001)and CG(dominant model OR=1.79,95%CI:1.15-2.79,P=0.0094)was observed when compared to the C group.Moreover,the presence of TLR9-1237 TC/CC+TLR9-1486 CC genotypes potentiate the risk for this neoplasm(OR=18.57;95%CI:5.06-68.15,P<0.0001).The TLR9 mRNA level was significantly higher in the GC group(RQ=9.24,P<0.0001)in relation to the CG group(RQ=1.55,P=0.0010)and normal mucosa(RQ=1.0).When the samples were grouped according to the polymorphic genotypes and the presence of H.pylori infection,an influence of TLR9-1237 TC+CC polymorphic genotypes(P=0.0083)and H.pylori infection(P<0.0001)was observed on the upregulation of mRNA expression.CONCLUSION Our findings show that TLR9 rs5743836 and rs187084 polymorphisms are associated with a higher risk of carcinogenesis gastric,and that TLR9 mRNA levels can be modulated by TLR9-1237 TC+CC variant genotypes and H.pylori infection.

Estrogen receptor gene polymorphism in a Chinese population with multiple sclerosis

This study sought to elucidate the role of the Pvull and Xbal polymorphisms of the estrogen receptor gene in 74 Chinese patients with multiple sclerosis, and 95 ethnicity-matched controls, using polymerase chain reaction-restriction fragment-length polymorphism analysis. The results revealed that the P allele of Pvull was significantly more prevalent in multiple sclerosis patients compared with controls （P = 0.019）. While distribution frequencies were significantly increased in female multiple sclerosis patients compared with female controls （P = 0.044）, no significant difference was observed between male patients and controls （P〉 0.05）. Frequencies of Ppxx genotypes were significantly higher in multiple sclerosis patients compared with controls （24.3% vs 12.8%, P = 0.025）. Genotypes and alleles of the estrogen receptor were not associated with age, number of attacks or expanded disability status scale scores of patients with multiple sclerosis. These findings indicate that the Pvull but not the Xbal polymorphism in the estrogen receptor gene is associated with susceptibility to multiple sclerosis in the Chinese population. In addition, women with P allele appear to be particularly susceptible to multiple sclerosis.

Relationship between estrogen receptor gene polymorphism and clinical indexes associated with coronary heart disease

Objective: To investigate the relationship between estrogen receptor(ER) gene and the clinical indexes associated with coronary heart disease (CHD). Methods: By means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), we analyzed ER gene polymorphism in 84 CHD patients and 61 healthy subjects and non-CHD inpatients. The clinical indexes associated with CHD were analyzed in relation to the three ER genotypes. Results: There were significant differences in the incidence of hypertension (58.62%), fibrinogen (Fib) concentration (3.5±0.8 g/L), body mass index (BMI, 25.1±3.2), HDL-C concentration (1.0±0.2 mmol/L) between PP genotype group and other genotype groups (P<0.05). Conclusion: ER gene polymorphism may affect ER-mediated cardiovascular protective effect by modulating the expression of ER.

SNP Identification in α_(2A)-Adrenergic Receptor Gene in Chinese and the Effect on Gene Expression

Objective: To scan single nucleotide polymorphism ( SNP ) in Chinese alpha-2Aadrenergic receptor (α_(2A)-AR) gene and study the effects of the SNP on the gene expression.Methods: The complete sequence of α_(2A)-AR gene was analyzed with automated DNA sequencer to scanSNPs. Genomic DNA was extracted from whole blood and a 239 bp fragment containing the G/Cpolymorphism was amplified with PCR using a pair of. specific primers. PCR-RFLP was used to performthe genotyping of the SNP at the site-1 296 bp of the people in the North of China. Electrophoresismobility shift assay ( EMSA ) was used to study the binding of the 390 bp fragments (- 1 414-1 025bp) with G or C at the site-1 296 bp and nuclear extracts . Results: In our study, two SNPs werefound in α_(2A)-AR gene. Allele frequencies of the SNP at the site-1 296 bp were 0.61 and 0.39 forG and C , and the genotype frequencies were 0.34 , 0.54 and 0.13 for GG, GC and CC respectively fromthe people in the North of China. In the EMSA, a specific binding appeared in the complex ofnuclear extracts and DNA with C at-1 296 bp . Conclusion: Two SNPs exist in α_(2A)-AR gene from thepeople in the North of China , and DNA fragment with allele C of the SNP at the site-1 296 bp couldbind with a specific protein, which could influence the gene expression.

TLR2 and TLR4 polymorphisms influence m RNA and protein expression in colorectal cancer

AIM: To evaluate the effect of promoter region polymorphisms of toll-like receptor(TLR)2-196 to-174 del and TLR4-1607T/C(rs10759932) on m RNA and protein expression in tumor tissue and of TLR4+896A/G(rs4986790) on colorectal cancer(CRC) risk.METHODS: The TLR2-196 to-174 del polymorphism was investigated using allele-specific polymerase chain reaction(PCR) and the TLR4-1607T/C and TLR4+896A/G by PCR-restriction fragment length p o l y m o r p h i s m( R F L P). W e g e n o t y p e d 4 3 4 D N A samples from 194 CRC patients and 240 healthy individuals. The m RNA relative quantification(RQ) was performed in 40 tumor tissue samples by quantitative PCR Taq Man assay, using specific probes for TLR2 and TLR4 genes, and ACTB and GAPDH reference geneswere used as endogenous controls. Protein expression was analyzed by immunohistochemistry with specific primary antibodies.RESULTS: No association was found for TLR4-1607T/C and TLR4+896A/G by three statistical models(logadditive, dominant and recessive). However, based on dominant and log-additive models, the polymorphic variant TLR2-196 to-174 del was associated with increased CRC risk [dominant: odds ratio(OR) = 1.72, 95%CI: 1.03-2.89; P = 0.038 and log-additive: OR =1.59, 95%CI: 1.02-2.48; P = 0.039]. TLR2 m RNA expression was increased in tumor tissue(RQ = 2.36) when compared to adjacent normal tissue(RQ = 1; P < 0.0001), whereas the TLR4 m RNA showed a basal expression(RQ = 0.74 vs RQ = 1, P = 0.452). Immunohistochemistry analysis of TLR2 and TLR4 protein expression was concordant with the findings of m RNA expression. In addition, the TLR2-196 to-174 del variant carriers showed m RNA relative expression 2.19 times higher than wild-genotype carriers. The TLR2 protein expression was also higher for the TLR2-196 to-174 del variant carriers [117 ± 10 arbitrary unit(a.u.) vs 95 ± 4 a.u., P = 0.03]. However, for the TLR4-1607T/C polymorphism no significant difference was found for both m RNA(P = 0.56) and protein expression(P = 0.26).CONCLUSION: Our findings suggest that TLR2-196 to-174 del polymorphism increases TLR2 m RNA expression and is associated with higher CRC risk, indicating an important role in CRC genetic susceptibility.

Characterization and strong risk association of TLR2 del-196 to-174 polymorphism and Helicobacter pylori and their influence on mRNA expression in gastric cancer

BACKGROUND Toll-like receptor-2(TLR2) is responsible for recognizing Helicobacter pylori(H.pylori) and activating the immune response. Polymorphisms in TLR2 may modulate gastric carcinogenesis.AIM To evaluate whether the TLR2 19216 T/C(rs3804099) and TLR2-196 to-174 ins/del(rs111200466) polymorphisms contribute to gastric carcinogenesis in the Brazilian population, and to determine the influence of both polymorphisms and H. pylori infection on TLR2 mRNA expression.METHODS DNA was extracted from 854 peripheral blood leukocyte or gastric tissue samples[202 gastric cancer(GC), 269 chronic gastritis(CG), and 383 control/healthy(C)]and genotyped by allele-specific PCR or restriction fragment length polymorphism(RFLP)-PCR. Quantitative polymerase chain reaction by Taq Man■ assay was used to quantify TLR2 mRNA levels in fresh gastric tissues(48 GC, 36 CG, and 14 C).RESULTS Regarding the TLR2-196 to -174 polymorphism, the ins/del and del/del genotypes were associated with a higher risk of GC by comparison with the C in all of the analyzed inheritance models(codominant, dominant, recessive, overdominant and log-additive;P < 0.0001). Similarly, an increased risk was observed when comparing the GC and CG groups [codominant(P < 0.0001), dominant(P <0.0001), recessive(P = 0.0260), overdominant(P < 0.0001) and log-additive(P <0.0001)]. In contrast, TLR2 19216 T/C was associated with a protective effect in the GC group compared to the C group [dominant(P = 0.0420) and log-additive(P =0.0300)]. Regarding the association of polymorphisms with H. pylori infection,individuals infected with H. pylori and harboring the TLR2-196 to-174 ins/del polymorphism had an increased risk of gastric carcinogenesis [codominant(P =0.0120), dominant(P = 0.0051), overdominant(P = 0.0240) and log-additive(P =0.0030)], while TLR2 19216 T/C was associated with a protective effect[codominant(P = 0.0039), dominant(P < 0.0001), overdominant(P = 0.0097) and log-additive(P = 0.0021)]. TLR2 mRNA levels were significantly increased in the GC group(median RQ = 6.95) compared to the CG group(RQ = 0.84, P < 0.0001)and to the normal mucosa group(RQ = 1.0). In addition, both H. pylori infection(P < 0.0001) and the presence of the polymorphic TLR2-196 to -174 del(P = 0.0010)and TLR2 19216 C(P = 0.0004) alleles influenced TLR2 mRNA expression.CONCLUSION The TLR2-196 to-174 ins/del and TLR2 19216 T/C polymorphisms are strongly associated with GC. TLR2 mRNA expression levels are upregulated in neoplastic tissues and influenced by both the presence of H. pylori and variant genotypes.

Dialogue between estrogen receptor and E2F signaling pathways: The transcriptional coregulator RIP140 at the crossroads

Estrogen receptors and E2F transcription factors are the key players of two nuclear signaling pathways which exert a major role in oncogenesis, particularly in the mammary gland. Different levels of dialogue between these two pathways have been deciphered and deregulation of the E2F pathway has been shown to impact the response of breast cancer cells to endocrine therapies. The present review focuses on the transcriptional coregulator RIP140/NRIP1 which is involved in several regulatory feed-back loops and inhibitory cross-talks between different nuclear signaling pathways. RIP140 regulates the transactivation potential of estrogen receptors and E2Fs and is also a direct transcriptional target of these transcription factors. Published data highlight the complex regulation of RIP140 expression at the transcriptional level and its potential role in transcription cross-talks. Indeed, a subtle regulation of RIP140 expression levels has important consequences on other transcription networks targeted by this coregulator. Another level of regulation implies titration mechanisms by which activation of a pathway leads to sequestration of the RIP140 protein and thus impinges other gene regulatory circuitries. Altogether, RIP140 occupies a place of choice in the dialogue between nuclear receptors and E2Fs, which could be highly relevant in various human pathologies such as cancer or metabolic diseases.

Estrogen receptor alpha gene polymorphism associated with type 2 diabetes mellitus and the serum lipid concentration in Chinese women in Guangzhou

Background Estrogen might play an important role in type 2 diabetes mellitus pathogenesis. A number of polymorphisms have been reported in the estrogen receptor alpha （ERα） gene （also named ESR1）, including the XbaⅠ and PvuⅡ restriction enzyme polymorphisms of ESR1, which may be involved in disease pathogenesis. The aim of this study was to determine whether ER0t gene polymorphisms are associated with type 2 diabetes mellitus and serum lipid level. Methods Two hundred and ninety-nine patients with type 2 diabetes mellitus were compared with three hundred and forty-one health controls of Guangzhou in China, both were male and postmenopausal female residents at 51--70 years. ESR1 genotyping was performed using polymerase chain reaction （PCR） and PvulI and XbaI restriction fragment length polymorphism （PCR-RFLP） analysis. Results ESR1 allelic frequencies of P, p and X, x alleles were 0.408, 0.592; 0.360, 0.640 in the type 2 diabetes mellitus group and 0.318, 0.682; 0.328, 0.672 in the control group, respectively. In case-control study, there was significant difference in PvuⅡ, but not XbaⅠ, allele frequency between the type 2 diabetes mellitus and control groups （P=0.001 and P=0.122）. When the group was separated into men and women, the difference was significant in women （P〈0.001） but not in men （P=0.854） with the PvulI genotype, and the effect of PvulI variant on the development of type 2 diabetes mellitus was improved with aging. In addition, PvulI genotype was associated with blood glucose [fasting blood glucose （FBG）, postprandial blood glucose （PBG）] and serum lipid [total cholesterol （TC） and low density lipoprotein （LDL）-c] concentration in healthy women. Conclusions PvuII polymorphism of ESRI increases susceptibifity to type 2 diabetes mellitus in Chinese Guangzhou women. ESR1 variants may also impact serum lipid metabolism, which might provide a mechanism connecting ESR1 to type 2 diabetes.

Relationship between bone mineral density and polymorphism ofthe estrogenreceptor genein healthy postmenopausal women in China

Objective To investigate the possible relationship between bone mineral density and polymorphism of the estrogen receptor (ER) gene in Shanghai healthy postmenopausal women Methods 250 unrelated healthy postmenopausal women were selected for bone mineral density (BMD) determination by Dual energy X ray absorptiometry (DEXA) and polymorphism of estrogen receptor gene analyses by polymerase chain reaction restriction fragment length polymorphism (PCR RFLP) Results PvuⅡ polymorphisms of ER gene was associated with low Troch BMD ( P =0 0153) while there was no significant relationship between XbaⅠ polymorphism of ER gene and BMD at any of skeletal sites included in the present study, and the combination of PvuⅡ and XbaⅠ polymorphisms of ER gene was significantly associated with both low Lumbar 2-4 ( P =0 0369) and Troch ( P =0 0384) BMD Multiple stepwise regression analysis also indicated that two combined polymorphisms were correlated significantly with Lumbar 2-4 BMD ( P =0 0254) while this correlation was not revealed at any other skeletal sites Conclusion There is significant relationship between the polymorphism of ER gene and both Lumbar 2-4 BMD and Troch BMD It is significant to explore the pathogenesis of osteoporosis and to prevent the development of osteoprosis by use of molecular

Association of estrogen receptor alpha gene polymorphisms with bone mineral density： a meta-analysis

Background A number of studies have examined the association between estrogen receptor alpha （ESR-a） gene polymorphisms and bone mineral density （BMD）, but previous studies of ESR-a gene Xbal （rs9340799） and Pvull （rs2234693） polymorphisms have been hampered by small sample size, regional restrictions and inconclusive results. Thus a meta-analysis is needed to assess their pooled effects. Methods This study reviewed all published articles indexed in Pubmed using the keywords in the title or abstract. All data were extracted independently by two reviewers using a standard form, the studies were meta-analyzed and minor discrepancies were resolved by authors＇ discussion. Results Twenty seven eligible studies involving 8467 women and 2032 men were identified. The Xbal and Pvull polymorphisms were significantly associated with BMD of the lumbar spine. XX and PP homozygotes had a protective effect in comparison with carriers of the x and p alleles, the effects were more significant in premenopausal women or Western women. At the femoral neck, the results were different. XX served as a protective factor in postmenopausal women, Western women, Western postmenopausal women, and men, while PP was likely to serve as a risk factor in Eastern women, Eastern postmenopausal women, and men. Conclusions The Xbal polymorphism is correlated to BMD at diverse skeletal sites. PP had a protective role for the lumbar spine but might be a risk factor for the femoral neck.

Estrogen receptor gene polymorphisms and bone mineral density in Chinese postmenopausal women

Objective To investigate the relationships between the polymorphisms of estrogen receptor (ER) gene, bone mineral density (BMD) and bone biochemical markers in Chinese postmenopausal women. Methods BMD of lumbar spine and femoral neck were measured using dual-energy X-ray absorptiometry (DEXA)in 186 Chinese postmenopausal women. The PvuⅡ and XbaⅠ polymorphisms of the ER gene were detected using polymerase chain reaction (PCR). Bone biochemical markers, serum alkaline phosphatase, osteocalcin and pyridinoline were measured by ELISA. Results The femoral neck(FN) BMD (Z score) was higher in pp compared to Pp (-0.01±0.12 vs. -0.35±0.09, P<0.05) while lumbar spine BMD (Z score) was higher in XX type compared to Xx and xx genotypes (0.01±0.45 vs -1.53±0.17, -1.29±0.10, P<0.001 and 0.001, respectively). Women without Px haplotype (n=79) had a higher BMD Z-score for the lumbar spine (-1.03±0.14 vs -1.45±0.11, P<0.05) and femoral neck (-0.01±0.11 vs -0.31±0.09, P<0.05) than those who had it (n=107). Conclusions The present study suggested that the pp and XX genotypes of ER gene might play a certain role in maintaining FN and lumbar spine BMD. ER genotypes without Px haplotype might be favorable to bone mass, while those with it might exert some harmful effect on bone mineral density.

Association of Estrogen Receptor Gene Polymorphisms and Primary Biliary Cirrhosis in a Chinese Population： A Case- Control Study

Background： Primary biliary cirrhosis （PBC） is a chronic and slowly progressive cholestatic liver disease characterized by destruction of the interlobular bile ducts and a striking female predominance, The aim of this study was to identity associations between estrogen receptor （ESR） gene polymorphisms with the risk of developing PBC and abnormal serum liver tests in a Chinese population. Methods： Thirty-six patients with PBC （case group） and 35 healthy individuals （control group） from the First Hospital of Jilin University were studied. Whole genomic DNA was extracted from all the participants. Three single-nucleotide polymorphisms （rs2234693~ rs2228480, and rs3798577） from ESR1 and two （rs1256030 and rs1048315） from ESR2 were analyzed by a pyrosequencing method. Demographic data and liver biochemical data were collected. Results： Subjects with the T allele at ESR2 rs1256030 had 1.5 times higher risk of developing PBC than those with the C allele （odds ratio [OR] = 2.1277, 95% confidence interval [CI] = 1.1872-4.5517）. Haplotypes TGC of ESRI rs2234693, rs2228480, and rs3798577 were risk thctors for having PBC. The C allele at ESRI rs2234693 was associated with abnormal alkaline phosphatase （OR 5.2469, 95% CI = 1.3704-20.0895） and gamma-glutamyl transferase （OR = 3.4286, 95% CI = 1.0083-13.6578） levels in PBC patients. Conclusions： ESR2 rs1256030 T allele may be a significant risk factor for the development of PBC. Screening for patients with gene polymorphisms may help to make early diagnoses in patients with PBC.

Estrogen receptor a gene PvulI polymorphism and coronary artery disease： a meta-analysis of 21 studies

The association between the estrogen receptor a gene （ESR1） Pvull polymorphism （c.454-397T〉C） and coronary artery disease （CAD） is controversial. Thus, we conducted a meta-analysis to evaluate the relationship. Data were collected from 21 studies encompassing 9926 CAD patients and 16710 controls. Odds ratios （ORs） with 95% confidence intervals （CIs） were used to assess the relationship between Pvull polymorphism and CAD. The poly- morphism in control populations in all studies followed Hardy-Weinberg equilibrium. We found a significant association between ESR1 Pvull polymorphism and CAD risk in all subjects. When the data were stratified by region, a significant association between ESR1 Pvull polymorphism and CAD risk was observed in Asian populations but not in Western populations. The current study suggests that ESR1 Pvull polymorphism has an important role in CAD susceptibility.

Correlation of estrogen receptor alpha gene polymorphisms and bone mineral density in Chinese women with chronic periodontitis

Background Periodontitis and osteoporosis are one of the frequently encountered diseases in post-menopausal women. Estrogen receptors （ERs） regulated bone metabolism. To investigate the possible effect of ER-elpha （a） gene polymorphisms on bone mineral density （BMD） in pre- and post-menopausal Chinese women with chronic periodontitis （CP）, we provided sufficient quantitative information concerning the correlation between ER gene polymorphisms and BMD in periodontitis. Methods Sixty-five post-menopausal and eighty pre-menopausal CP women, and sixty post-menopausal healthy individuals were recruited in this study. Genomic DNA was extracted from oral mucosa swab sample of each subject by the Chelex-100 method. Determination of the ER-a polymorphisms was performed by polymerese chain reaction-restriction fragment length polymorphism （PCR-RFLP） technique with Xbal and Pvull enzyme. The index for periodontal examination includes clinical attachment loss （CAL） and probing pocket depth （PPD）. BMD was measured by dual-energy X-ray absorptiometry （DEXA）. Results There were no significant differences between the ER-α genotypes of Pvull and Xbal and BMD in post-menopausal and pre-menopausal CP patients, respectively （P 〉0.05）. However, there was association between pre- and post-menopausal CP patients at BMD of lumbar spine L2-L4 （P=0.027） and Ward＇s BMD （P=0.004）. Furthermore, the post-menopausal CP women who carried Pvull TT genotype presented significantly lower Ward＇s BMD than the pre-menopausal CP women （P=-0.007）, meanwhile, the post-menopausal CP women who carried Xbal AA genotype presented significantly lower spine L2-L4 BMD than the pre-menopausal CP women （P=0.003）. Conclusions ER-α gene polymorphisms may be a susceptible indicator for BMD variation of lumbar spine L2-L4 and Ward in Chinese pre- and post-menopausal women patients with CP.

孤雌卤虫雌激素相关受体基因(ERR)的表达特性分析

雌激素通过其相关受体进而影响动物的生殖发育与成熟。为了探究雌激素相关受体基因ERR在孤雌卤虫卵巢发育和成熟过程中发挥的作用,本研究对孤雌卤虫(Artemia parthenogenetica)的ERR基因进行了克隆及相关生物信息学分析,并对该基因在不同组织(卵巢、头部、躯干)和卵巢在不同发育时期〔卵黄发生早期(early oocytes,EO)、卵黄发生晚期(late oocytes,LO)、早期胚胎(early embryos,EE)、晚期胚胎(later embryos,LE)〕的表达特征进行了探究。结果显示,ERR基因的开放阅读框(ORF)长1536 bp,共编码521个氨基酸,编码蛋白质的相对分子质量和等电点分别为5.7114×10^(4)和5.06,该蛋白质具有两个结构域,没有信号肽和跨膜结构。蛋白质的二级结构主要以无规则卷曲(46.97%)和α-螺旋(40.7%)为主,三级结构与二级结构的结果一致。在NJ系统进化树中,孤雌卤虫(A.parthenogenetica)与大型溞(Daphnia magna)和苏拉威西秀体溞(Diaphanosoma celebensis)最为相近,而与湿木白蚁(Zootermopsis nevadensis)、台湾乳白蚁(Coptotermes formosanus)的亲缘关系较远。表达特征结果显示,ERR基因在孤雌卤虫头部组织中的表达量显著高于卵巢组织和躯干组织中的表达量,且随着卤虫卵巢的发育,表达量逐渐上升,在EE达到最高,随后在LE显著下降(P<0.01)。

雌激素受体基因多态性与复发性流产的相关性研究

目的 探讨雌激素受体 (ER)基因多态性与复发性流产的相关性。方法 取实验组与对照组妇女外周血 ,采用聚合酶链式反应 限制性片段长度多态性 (PCR RFLP)方法研究ER基因的XbaⅠ和PvuⅡ酶切多态性 ,通过统计学分析探讨ER基因的多态性分布与复发性流产的关系。结果 ER基因PvuⅡ酶切多态性分布与复发性流产存在显著相关性 (P <0 .0 1) ,而ER基因XbaⅠ酶切多态性分布与复发性流产无显著相关性 (P >0 .1)。结论 ER基因PvuⅡ酶切多态性分布与复发性流产显著相关 (P <0 .0 1)。

雌激素受体基因多态性与女性绝经后骨质疏松症中医辨证分型关系的研究

目的 :研究中国绝经后女性雌激素受体基因多态性与绝经后骨质疏松症中医辨证分型的关系。方法 :对 2 4 6名中国绝经后女性 (年龄 4 4～ 80岁 ,平均 65 8岁 ) ,用分子生物学的方法分析内切酶PvuⅡ、XbaⅠ限制性长度片段多态性 (RFLPs) ,运用双能X线骨吸收法分别测其腰椎 (L1～ 4 )和股骨 (粗隆间、股骨颈、Ward′s区 )骨密度 ,根据中医虚证辨证分型标准 ,将研究对象分为肾阴虚型、肾阳虚型和阴阳俱虚型 ,观察雌激素受体基因多态性与骨密度及中医辨证分型的关系 ,RFLPs用Pp(PvuⅡ )和Xx(XbaⅠ )来表示 ,限制性部分缺失者用大写字母表示 ,存在者用小写字母表示。结果 :PPxx基因型 ( 2 1例 )骨密度Z score明显低于其他基因型 ( 2 2 5例 ) ,腰椎 ( - 0 71± 0 4 6) g/cm2 ,粗隆间 ( - 0 31± 0 58) g/cm2 ,股骨颈 ( - 0 84±0 66) g/cm2 ,Ward′s区 ( - 0 96± 0 85) g/cm2 ,该基因型女性中医辨证属阴阳俱虚型。结论 :雌激素受体基因RFLPs与中医辨证分型有关。

雌激素受体基因多态性在中国汉族人群中的分布特点

目的：了解雌激素受体（ＥＲ）基因多态性在中国正常人群中的分布及其在不同种族中分布的差异。方法：通过ＰＣＲ限制性酶切方法对１７２名正常个体的基因组ＤＮＡ中ＥＲ基因ＲＦＬＰ多态性进行了观察，并结合文献进行了不同种族间的分析比较。结果：中国正常人群中ＥＲ基因型以ｐｐ型，ｘｘ型和ｐｐｘｘ型发生频率最高，这种分布特点在正常男女间差异不显著。与其他种族相比较，ＥＲ基因型在中国正常人群中的分布与日本人群中的分布较为接近，而与瑞典，美国人群中的分布差异显著。结论：ＥＲ基因ＲＦＬＰ基因多态性在不同种族间分布存在着明显的差异。

雌激素受体基因多态性与乙型肝炎肝硬化的相关性研究

目的:探讨雌激素受体基因的PvuⅡ,XbaⅠ多态性与乙型肝炎肝硬化的关系,从基因水平上进一步探讨肝硬化的发病机制。方法:采用聚合酶链反应(PCR)-限制性片段长度多态性(RFLP)方法检测并比较了98例乙型肝炎肝硬化患者,72例慢性乙型肝炎患者,84例健康对照组的雌激素受体基因PvuⅡ和XbaⅠ多态性。结果:乙型肝炎肝硬化患者的雌激素受体基因的PvuⅡ多态性的Pp基因型和P等位基因频率明显高于健康对照组和慢性乙型肝炎组,pp基因型和p等位基因频率明显低于健康对照组和慢性乙型肝炎组,差异有统计学意义(P<0.05),PP+Pp基因型患肝硬化的风险是pp基因型的2.23倍(OR=2.23)。雌激素受体基因的XbaⅠ多态性分布在各组间比较,差异均无统计学意义(P>0.05)。结论:雌激素受体基因Pp基因型和P等位基因可能是肝硬化发病的遗传易感基因,pp基因型和p等位基因可能是肝硬化发病的保护基因。

健胎液对米非司酮处理的小鼠子宫雌孕激素受体基因表达的影响

目的探讨健胎液改善着床障碍小鼠子宫内膜容受性的分子机制。方法利用米非司酮诱导胚泡着床障碍小鼠模型 ,予健胎液进行干预 (中药组 ) ,于妊娠第 8天处死小鼠 ,采用蛋白印迹 (Westernblot)、逆转录聚合酶链式反应 (semi quantitativereversetranscriptasepolymerasechainreaction ,RT PCR)检测子宫雌、孕激素受体 (estrogenreceptor,ER ;progesteronereceptor,PR)蛋白及其基因表达。 结果中药组ER、PR蛋白及基因表达均较模型组显著提高 (P <0 0 5 ) ,与正常组比较差异无显著性 (P >0 0 5 )。结论健胎液通过调节胚泡着床障碍小鼠子宫组织ER、PR蛋白及基因的表达 ,促进子宫内膜发育 。