Porcine growth differentiation factor 9 gene polymorphisms and their associations with litter size

Growth differentiation factor 9 （GDF9） is expressed in oocytes and is thought to be required for ovarian folliculogenesis. Given this function, GDF9 may be considered as a candidate gene controlling pig ovulate rate. In this study, the complete coding sequence was cloned （encoding a 444 amino acid）, intron sequence and partial 5＇-UTR of pig GDF9. RT-PCR results showed that GDF9 mRNA is expressed in a wide range of tissues of the ruttish Erhualian pig. The expression levels of GDF9 mRNA in pituitary, ovary, uterus and oviduct are higher in the Erhualian pigs than those in Duroc pigs, especially in pituitary with a significant difference （P 〈 0.05）. Comparative sequencing revealed 12 polymorphisms, including 8 single nucleotide polymorphisms （SNPs） and one 314 bp indel in noncoding regions, and the other 3 SNPs in coding regions. Four polymorphisms, G359C, C1801T, T1806C and 314 bp indel, were developed as markers for further use in population variation and association studies. The G359C polymorphism segregates only in Chinese native pigs, Erhualian and Dahuabai, on the contrary, 314 bp indel segregates only in Duroc and Landrace. C1801T and T1806C sites seem to be completely linked and segregate in Erhualian, Dahuabai and Landrace. In a word, GDF9 may be not associated with pig litter size in extensive populations as per the studies of allele distributions of the four polymorphisms and pilot association in four breeds.

Toll-like receptor 9 gene mutations and polymorphisms in Japanese ulcerative colitis patients

Abnormal innate immune responses toward luminal bacteria play an important role in the pathogenesis of inflammatory bowel disease.It has been demonstrated that bacteria having CpG DNA ameliorate experimental colitis in mice,and Toll-like receptor 9 (TLR9) signaling mediates the anti-inflammatory effects in mouse colonic inflammation.A gene variation in NOD2/CARD15 has been reported in Crohn's disease (CD) patients in Western countries,but this variation has not been identified in Japanese CD patients.Therefore,we hypothesized that TLR9 is a key factor in the development of ulcerative colitis (UC),and we investigated gene mutations and polymorphisms of TLR9 in Japanese UC patients.Three single nucleotide polymorphisms (SNPs) in TLR9 were identified in healthy controls,and were assessed in 48 UC patients and 47 healthy controls.Control subjects were matched for age,sex and date of blood sampling from among a subgroup of participants.We found that TLR9-1486CC,1174GG and 2848AA increase the risk of UC [odds ratio (OR) 2.64,95% confidence interval (95% CI):1.73-6.53,P=0.042],and TLR9-1486TT,1174AA and 2848GG decrease the risk of UC (OR 0.30,95% CI:0.10-0.94,P=0.039),although there were no correlations between SNPs and disease phenotype or TLR9 mRNA expression.These findings suggest that TLR9 polymorphisms are associated with increased susceptibility to UC.

The association between matrix metalloprotease-9 gene polymorphisms and primary angle-closure glaucoma in a Chinese Han population

AIM:To examine the association between the single nucleotide polymorphisms (SNPs)of matrix metalloprotease-9 (MMP-9) gene and primary angleclosure glaucoma(PACG)in a Chinese Han population.METHODS:DNA samples were extracted from peripheral-blood mononuclear cells of 214 PACG patients and 224 healthy controls.Genotyping of rs3918249,rs3918254,rs17577 and rs3787268 in MMP-9was performed using polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP)analysis and the direct sequencing technique.The association between these genetic polymorphisms and risk of PACG was estimated by χ2 test.RESULTS:The distributions of rs3918249,rs3918254,rs17577 and rs3787268 genotypes among cases and healthy controls were compatible with that from HardyWeinberg equilibrium(HWE,P】0.05).The increased frequency of CC and CT genotypes of rs3918254 were observed in PACG patients compared to healthy controls[P=0.006,P corrected(Pcorr)=0.048].The haplotype analysis showed that the CCGG haplotype was nominal associated with PACG(P=0.015),however,the significant was lost when the Bonferroni correction was used(Pcorr=0.105).CONCLUSION:Our results revealed that rs3918254 in MMP-9 may be a susceptible locus to PACG in China,people with the CC and CT genotypes of rs3918254 are more susceptible to PACG.The susceptibility to PACG inChinese Han patients may be not influenced by SNPs rs3918249,rs3787268 and rs17577 in MMP-9.

Matrix metalloproteinase-9-1562C>T polymorphism may increase the risk of lymphatic metastasis of colorectal cancer

AIM. To explore the role of the matrix metalloproteinase-9 （MMP-9） polymorphism in colorectal cancer （CRC） in a northeast Chinese population.METHODS： Genotyping of MNP-9-1562C〉T and 279R〉Q polymorphisms was carried out on blood samples from 137 colorectal cancer patients and 199 controls using polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. Multivariate logistic regression models were used to calculate adjusted odds ratios （OR） and 95% confidence intervals （95% CI）.RESULTS： The distribution of IVllVlP-9 -2562C〉T and 279 R〉Q genotype was not significantly associated with the risk of CRC. However, the risk of Ilymph node metastasis of CRC was increased in patients with the -1562T allele （OR = 2.601; 95% CI = 1.160-5.835; P = 0.022）. The frequency of MMP-9 279RR ＋ RQ genotype was higher than the QQ genotype among CRC patients younger than sixty years old （OR = 0.102, 95% CI = 0.013-0.812; P = 0.012）.CONCLUSION： Our results indicated that the MMP-9- 1562C〉T polymorphism affects lymph node metastasis of CRC. In addition, the MMP-9 279R allele may lead to a younger age of onset of colorectal cancer.

E670G polymorphism of PCSK9 gene of patients with coronary heart disease among Han population in Hainan and three provinces in the northeast of China

Objective: To investigate the correlation between E670 G polymorphism of proprotein convertase subtilisin/kexin type 9(PCSK9) gene and coronary heart disease(CHD), and contrastively study the regional differences of E670 G polymorphism of PCSK9 gene between patients with CHD among the Han population in Hainan and three provinces in the northeast of China(TPNC), providing scientific basis for prevention and treatment of patients with CHD in different regions. Methods: A total of 233 cases of patients with CHD were selected from the Han population in Hainan and TPNC as the experimental group(118 cases from Hainan, 115 cases from TPNC), and 239 cases with non-CHD were selected among the Han population also in the two regions as control group(125 cases from Hainan, 114 cases from TPNC). The triglyceride(TG), total cholesterol(TC), high density lipoprotein cholesterol and low density lipoprotein cholesterol(LDL-C) levels of plasma were tested and PCR-RFLP method was used to test the E670 G polymorphism of PCSK9 gene. The statistical software package SPSS 21.0 was used for the statistical analysis and P<0.05 was considered as statistically significant. Results: The levels of systolic pressure, diastolic blood pressure, fasting blood sugar, TC, TG, and LDL-C of patients in CHD group were significantly higher than those in non-CHD group, while the high density lipoprotein cholesterol level was lower than that in non-CHD group(P<0.05). In CHD group, the frequencies of AG, GG genotypes of PCSK9 gene and G allele were higher than those in non-CHD group(P<0.05), and in CHD group, the frequencies of AG, GG genotypes and G allele of patients both in Hainan and TPNC were higher than those in control group(P<0.05). Among the patients with CHD, the frequencies of GG genotype and G allele of patients in Hainan were lower than those in TPNC(P<0.05), and in CHD group, the levels of TG, TC and LDL-C of GG genotype were higher than those of AA genotype(P<0.05). While in non-CHD group, there were no significant differences between the frequencies of GG genotype and G allele of patients in Hainan and TPNC(P>0.05). Conclusions: There was a close correlation between the E670 G polymorphism of PCSK9 gene and CHD with serum lipid level. Among Han population in Hainan and TPNC, the E670 G polymorphism of PCSK9 gene of patients with CHD exhibited regional differences.

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.

Identification of Novel Polymorphisms in Porcine Ring Finger Protein 4 and Matrix Metalloproteinase 9 Genes and Association Analysis with Litter Size Traits

Reproduction trait plays an important role in pig production. Identification of molecular markers that are linked to litter size may contribute to the genetic development of porcine reproduction traits. In this study, porcine ring finger protein 4 （pRNF4） and Matrix metalloproteinase 9 （pMMP-9） were selected as candidate genes on the basis of their physiological roles in reproduction. Two single-nucleotide polymorphisms （416C〉T in pRNF4 and - 1257G〉A in pMMP-9） that could be detected by PCR restriction fragment length polymorphism （PCR-RFLP） were discovered and tested for statistical associations with litter size traits in three populations. For 416C〉T, TT genotype was associated with a significantly higher （p〈0.05） number of live births than those recorded for CC sows and the additive effect was significant （p〈0.05） in Qingping and Min Pigs populations in later parities. For -1257G〉A, inconsistent results were found in three populations. The results suggested that T allele in pRNF4 gene might confer a high prolificacy in breeding and further studies were needed to confLrm the results.

The proprotein convertase subtilisin/kexin type 9 geneE670G polymorphism and serum lipid levels in the Guangxi Bai Ku Yao and Han populations

Background The association of E670G polymorphism in the proprotein convertase subtilisin/kexin type 9 (PCSK9) gene and serum lipid profiles is inconsistent in dif- ferent ethnic groups.Bai Ku Yao is a special subgroup of the Yao minority in China.The present study was undertaken association of PCSK9 E670G polymorphism and several environmental factors with serum lipid levels in the Guangxi Bai Ku Yao and Han populations.Methods A total of 649 subjects of Bai Ku Yao and 646 participants of Han Chinese were randomly selected from our previous stratified randomized cluster samples.Genotyping of the PCSK9 E670G polymorphism was performed by polymerase chain reaction and restriction fragment length polymorphism combined with gel electrophoresis,and then confirmed by direct sequencing. Results The levels of serum total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C) and apolipoprotein(Apo) AI were lower in Bai Ku Yao than in Han(P【0.01 for all).The frequency of A and G alleles was 98.00%and 2.00%in Bai Ku Yao,and 95.20%and 4.80%in Han(P【0.01);respectively. The frequency of AA,AG and GG genotypes was 95.99%,4.01%and 0%in Bai Ku Yao,and 91.02%, 8.36%and 0.62%in Han(P【0.01);respectively.There were also significant differences in the genotypic and allelic frequencies between n and the ratio of ApoAI to ApoB in Han Chinese but not in Bai Ku Yao were different between the AA and AG/GG genotypes(P【0.05 for all).The G allele carriers had higher serum HDL-C and higher ApoAI to ApoB ratio than the G allele noncarriers.When serum lipid parameters in Han were analyzed according to sex,the G allele carriers had higher serum HDL and ApoAI levels in males (P【0.05),and lower ApoB level and higher ApoAI to ApoB ratio in females(P【0.05 for all).Multiple linear regression analysis showed that serum HDL-C levels were correlated with genotypes in both ethnic groups(P【0.05 each).Serum lipid parameters were also correlated with sex,age,body massindex,alcohol consumption,cigarette smoking,and blood pressure in both ethnic groups(P【0.05-0.001).Conclusions These results suggest that the PCSK9 E670G polymorphism is mainly associated with some serum lipid parameters in the Han population,both gender show different relations to different serum lipid parameters.The G allele carriers might have higher serum lipid profiles than the G allele noncarriers. ormal LDL-C(≤3.20 mmol/L) and high LDL-C subgroups (】 3.20 mmol/L,P【0.01;respectively) in Bai Ku Yao, and between normal ApoB(≤1.14 g/L) and high ApoB subgroups(】 1.14 g/L,P 【 0.01;respectively) in Han.

Association between the-1562 C/T polymorphism in the MMP-9 promoter and phenotype of esophageal squamous cell carcinoma in northern Chinese population

Objective To conduct a case-control study on the association of the nucleotide polymorphisms in the promoter region of the matrix metalloproteinase-9(MMP-9)gene with phenotype of esophageal cancer.Methods All subjects were unrelated residents in northern regions of China.Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)analysis was used to determine the MMP-9 genotypes.Results The overall distribution of genotypes in the patients was not different from that in the controls(OR=0.77,95% CI=0.45-1.34;P=0.36).There were no significant differences between the patients and the control subjects in terms of the distributions of sex and age,smoking status,alcohol dependence,pickled diet status,or history of environmental exposure.The patients were further examined with stratifications by age,sex,grade,depth of tumor invasion,lymphatic invasion,venous invasion and TNM staging.The results showed no pronounced association among the stratifications.Conclusion There is no significant association between the MMP-9 single nucleotide polymorphism genotypes and phenotype of esophageal cancer.

Association of Matrix Metalloproteinase-9 and p53 Gene Polymorphisms with Genetic Susceptibility to No-small-cell Lung Cancer

Matrix metalloproteinase-9（MMP-9） and p53 genes play an essential role in the multi-step process of tumorigenesis in lung cancer. Single nucleotide polymorphisms（SNPs） of MMP-9 and p53 genes are associated with the risk and progression of many cancers. In this study, we evaluated the association of the R279Q polymorphism of MMP-9 or the A1/A2 polymorphism of p53 gcne with the risk of no-small-cell lung cancer（NSCLC） in Han population of Northeast China. We examined the frequency of SNPs in the two kinds of genes of 50 patients with NSCLC and 50 cancer-free controls frequency-matched by age and sex. Polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP） technique was used to determine the genotypes. The results indicate that the 279RR genotype in MMP-9 gene and the A1/A2 genotype in p53 gene show a significantly increased risk of NSCLC. Therefore, the MMP-9 279RR and p53 A1/A2 genotypes may be used as markers for susceptibility to NSCLC in Han population of Northeast China.

ABCB9 polymorphism rs61955196 is associated with schizophrenia in a Chinese Han population

BACKGROUND Schizophrenia(SCZ)is a complex disease which can be affected by both genetic and environmental factors.Prenatal famine exposure may cause changes in DNA methylation levels of genes.Meanwhile,maternal nutrition during pregnancy is a pivotal environmental factor in the development of SCZ.DNA methylation may be an intermediate factor mediating exposure to famine during pregnancy and SCZ,and DNA methylation quantitative trait loci might serve as a promising tool for linking SCZ and prenatal famine.AIM To analyze the association between prenatal famine exposure and SCZ risk in Northeast Han Chinese through analysis of DNA methylation related loci.METHODS A total of 954 Han Chinese from Northeast China were recruited,including 443 patients with SCZ and 511 healthy controls.The participants were further divided into famine(born in 1960-1962)and non-famine(born in 1963-1965)groups to investigate the effect of prenatal famine exposure.Four single-nucleotide polymorphisms(SNPs)selected according to the relevant literature were genotyped,namely,rs11917047 in PTPRG,rs2239681 in IGF2,rs3842756 in INSIGF,and rs61955196 in ABCB9.DNA were extracted from peripheral blood samples,and the genotypes of these SNP loci were detected using the improved Multiple Ligase Detection Reaction multiple SNP typing technique.The associations of the DNA methylation related SNPs with SCZ risk and prenatal famine,and their interactions were analyzed using logistic regression analysis and generalized multifactor dimensionality reduction(GMDR)software.RESULTS Based on the sequencing data,genotype distributions and allele frequencies of the four selected SNPs were determined.All genotype frequencies of the four SNPs in the healthy control group were tested for deviation from Hardy-Weinberg equilibrium(P>0.05).Logistic regression analysis showed that rs61955196 was significantly associated with SCZ risk in the log-additive model[odds ratio(OR):1.22;95%confidence interval(CI):1.01-1.48;P=0.040].We also found that the rs61955196 allele was related with an enhanced risk of SCZ(G>C,OR:1.22;95%CI:1.01-1.47;P=0.042).However,no associations were observed between rs11917047,rs2239681,or rs3842756 and SCZ risk.Under the optimal genetic model,no significant association of famine with the four SNPs was seen.Though the gene–gene interactions between rs2239681 and rs61955196 were found in GMDR analysis,none of the gene-gene interactions and gene-famine interactions were associated with the risk of SCZ.CONCLUSION Our study suggested that rs61955196 in ABCB9 is associated with SCZ susceptibility in Northeast Han Chinese,providing insight into genetic effects on SCZ.

Genetic Polymorphisms of CYP2C9: Comparison of Prevalence in the Lebanese Population with Other Populations

Background: There is little knowledge about genotyping of cytochrome P450s in the Middle East, and there has not been any report on the genotype of CYP 2C9 allelic variants in Lebanese population. Aims and objectives: The purpose of the study was to determine and compare the frequencies of the cytochrome P450 CYP2C9 variants in the Lebanese population with the frequencies in other ethnic populations. Methods: CYP2C9 genotypes were determined in a total of 146 samples of unrelated, healthy Lebanese individuals residing in different areas in Lebanon. Following DNA extraction from buccal cells and polymerase chain reaction, genotyping was performed by Pyrosequencing method. CYP2C9 genotypes results were compared to other populations;i.e., Middle Easterns, Europeans, Asians, and African Americans. Results and discussion: The frequencies of the CYP2C29*2, CYP2C9*3, and CYP2C9*4 alleles were 11.305%, 11.645%, and 1.025% respectively. No CYP2C9*5 allele variants were found among the Lebanese study sample. Vol- unteers could be divided into three CYP2C9 genotype groups: subjects (76.71%) with no mutated alleles (CYP 2C9*1*1;homozygous extensive metabolizers, EM), 21.23% with one mutated allele (CYP 2C9*1*2, *1*3, *1*4, and *1*5;heterozygous intermediate metabolizers IM), and 2.06% with two mutated alleles, homozygous variants as poor metabolizers, PM). The comparative analysis using genotype groups of different populations showed differences among Leba- nese and other Caucasians. Conclusion: This is the first report from Lebanon on CYP2C9 variants;it highlights a higher frequency of CYP2C9 extensive metabolizers compared to other populations including Caucasians. The results serve as a database on CYP 2C9 polymorphisms and baseline clinical data for dosing and avoiding adverse drug reac- tions of drugs metabolised by CYP2C9 in Lebanese patients.

Matrix Metalloproteinase-9 Promoter Polymorphism in Primary Spontaneous Pneumothorax

Primary spontaneous pneumothorax （PSP） is a common clinical problem occurring in apparently healthy subjects. However emphysema like changes （ELC） in patients with （PSP） has been demonstrated, which is associated with the degradation of extraceilular matrix （ECM）. MMP-9 has been proposed to play a role in the development of emphysema and is involved in the digestion of （ECM）. Among several polymorphic changes reported in MMP-9 regulatory region, the C-1562T polymorphism increases its promoter activity Aim of the work： To test the hypothesis that the MMP-9 C-1562T polymorphism has a role in the occurrence of PSP. Subjects and Methods： MMP-9 （C-1562T） genotypes of 104 PSP patients and 97 healthy subjects （control） were determined by the polymerase chain reaction, followed by a restriction fragment length polymorphism analysis. Results： This study showed that the genotype frequency for MMP-9 1562 C 〉 T differed significantly between healthy individuals and PSP patients, with higher T allele in MMP-9 promoter in PSP patients, and the C/T ＋ T/T genotypes significantly modified the risk of developing PSP, and were associated with increased risk for PSP. Conclusion： MMP-9-1562C/T genotype may be a potentially indicative factor for susceptibility to PSP, that is, genotypes with the T allele significantly increase the risk of development of PSP.

Combined effect of <i>CYP2C9</i>and <i>VKORC1</i>polymorphisms on warfarin maintenance dose in Omani patients

Excepting host genetic factors, other influences on the pharmacokinetic and pharmacodynamic behavior of warfarin are subject to variations during the treatment despite attempts to stabilize the INR. In 214 Omani patients on warfarin therapy, we evaluated the extent of influence of known genetic predictors of warfarin dose variability, namely CYP2C9, CYP4F2 and VKORC1 gene polymorphisms in a genetically heterogeneous patient population. When patients were stratified according to their daily warfarin maintenance dose (to maintain INR between 2 and 3) into “low dose” (sensitive), “medium dose” (intermediate) and “high dose” (resistance) groups, overall, seven patients with three or four mutant alleles fell in the sensitive group and consequently 25% (7 out of 28) of at risk patients for over anticoagulation can be recognized by prospective pharmacogenetic testing in this patient population. Pre-prescription genotyping of these loci prior to therapy initiation will therefore benefit a small fraction of this population.

不同致病性H9N2亚型禽流感病毒诱导鸡TLR-7和Mx基因mRNA转录水平的差异

本试验拟研究不同致病性H9N2亚型禽流感病毒诱导SPF鸡、SPF鸡胚和鸡胚成纤维细胞中TLR-7和Mx基因mRNA转录水平的动态变化,为不同致病性H9N2亚型禽流感病毒诱导IFN-Ⅰ产生差异的机制提供依据。150只6周龄SPF鸡随机分为3组,分别静脉接种PBS、105 EID50毒株SD/196和SD/818各0.2mL,分别于接种后的第1、3、5、7和10天每组剖杀8只,收集具有明显病变差异的组织;180枚10日龄SPF鸡胚分别尿囊腔接种PBS、104 EID50的SD/196毒株和SD/818毒株各0.1mL,分别于接种后的第4、8、16、24、32和48小时每组收集胚体10个;鸡胚成纤维细胞分别接种PBS、SD/196和SD/818毒株病毒液(1moi)进行37℃培养,于感染后的第4、8、16、24、32和48小时收集细胞;动态收集的病变组织、胚体和成纤维细胞通过荧光定量PCR测定TLR-7和Mx基因mRNA水平的表达。结果显示,肺、肾、十二指肠和法氏囊为主要病变差异组织,除鸡胚TLR-7的表达外,毒株SD/818大部分时间点诱导主要病变差异组织、鸡胚和成纤维细胞中TLR-7和Mx基因mRNA的表达显著高于毒株SD/196(P<0.05)。结果表明不同致病性H9N2亚型禽流感病毒明显诱导了SPF鸡主要病变差异组织、SPF鸡胚和鸡胚成纤维细胞中TLR-7和Mx基因mRNA的表达,且较高致病性毒株SD/818诱导表达量明显高于较低致病性毒株SD/196。

TLR-9 rs187084基因多态性与山东汉族人群系统性红斑狼疮的相关性研究

目的研究山东地区汉族人群TLR-9基因rs187084单核苷酸多态性,探讨其与系统性红斑狼疮易感性之间的关系。方法采用聚合酶链反应和限制性片段长度多态性等方法对92例系统性红斑狼疮患者和88名健康对照TLR-9基因rs187084C/T多态性进行分析,计算基因型和等位基因频率。结果(1)SLE患者TLR-9基因rs187084CC、CT、TT基因型频率分别是0.185、0.413和0.402,与对照组相比没有统计学差异(χ2=2.99,P>0.05);(2)SLE患者TLR-9基因rs187084C、T等位基因频率分别是0.394、0.606,与对照组间差异没有统计学差异(χ2=3.07,P>0.05)。结论山东汉族人群SLE的易感性与TLR-9基因rs187084多态性无关。

小鼠视神经损伤后视网膜中TLR-9与MyD88表达变化

目的:探讨Toll样受体-9(TLR-9)与髓样分化因子(MyD88)在小鼠视神经损伤(ONI)后视网膜中的表达变化。方法:选取8周大小的雄性C57BL/6J小鼠36只,随机分为6组:空白对照组(未做任何处理)、ONI 1d组(视神经损伤后1d取材)、ONI 3d组(视神经损伤后3d取材)、ONI 5d组(视神经损伤后5d取材)、ONI 7d组(视神经损伤后7d取材)、ONI 14d组(视神经损伤后14d取材)。分组后通过视神经夹持的方法制作小鼠视神经损伤模型,利用RT-qPCR与Western-blot检测各组小鼠视网膜中TLR-9与MyD88的mRNA和蛋白水平。结果:ONI 1d组视网膜中TLR-9与MyD88的mRNA和蛋白水平与空白对照组比较无差异(P>0.05);ONI 3d、ONI 5d、ONI 7d、ONI 14d组视网膜中TLR-9与MyD88的mRNA和蛋白水平与空白对照组比较均明显增加(P<0.01)。与空白对照组比较视网膜中TLR-9与MyD88的mRNA和蛋白水平在小鼠ONI 3d开始升高(P<0.01),ONI 5d达到峰值(P<0.001),ONI 7d开始逐渐下降(P<0.01)。结论:小鼠视神经损伤能够激活视网膜中TLR-9与MyD88表达,TLR-9与MyD88可能在视神经损伤的进程中起重要作用。

TLR-9基因多态性与系统性红斑狼疮的相关性研究

目的 分析山东地区汉族人群TLR-9基因rs187084、rs352139等位点的多态性,探讨其与系统性红斑狼疮的相关性.方法 采用PCR-RFLP等方法对92例系统性红斑狼疮患者和88例健康对照者TLR-9 基因rs187084、rs352139的多态性进行分析,计算基因型和等位基因频率结果：SLE患者TLR-9基因rs187084CC、CT、TT基因型频率分别是0.185、0.413和0.402,rs352139AA、AG、GG基因型频率分别是0.228、0.533和0.239,与对照组间差异没有统计学意义（χ2=2.99,P〉0.05,χ2=4.54,P〉0.05）.结论 TLR-9基因可能在SLE发病中起一定作用.

TLR-9在胰腺癌细胞化疗中对吉西他滨耐药性的影响

目的:探讨TLR9影响胰腺癌PANC-1细胞对吉西他滨的耐药性。方法:将PNCA-1细胞随机分为3组。A组给予生理盐水+吉西他滨,B组给予TLR9激动剂+吉西他滨,C组给予TLR9抑制剂+吉西他滨。检测3组细胞凋亡水平及bcl-2蛋白的表达。结果:B组凋亡率小于A组与C组,差异具有统计学意义(P<0.05),而C组凋亡率大于A组,差异具有统计学意义(P<0.05)。A组Bcl-2的表达与B组比较,差异具有统计学意义(P<0.01)。B组Bcl-2的表达与C组比较,差异具有统计学意义(P<0.01)。A组Bcl-2的表达与C组比较,差异具有统计学意义(P<0.05)。结论:激活TLR9可明显增加胰腺癌PNAC-1细胞株对吉西他滨化疗的耐药性,抑制TLR9可增加胰腺癌细胞对吉西他滨的敏感性。

系统性红斑狼疮患者IRF5、TLR-9基因表达的研究

目的 研究IRF5、TLR-9基因表达水平与系统性红斑狼疮的相关性.方法 收集92例SLE患者与88名正常对照人群的外周血,提取总RNA并反转录为cDNA,以实时荧光定量聚合酶链反应方法检测患者组和对照组的IRF5、TLR-9的mRNA表达水平.结果 SLE患者外周血IRF5、TLR-9 mRNA的表达分别是0.232±0.076、0.189±0.054,与对照组相比,SLE患者IRF5、TLR-9 mRNA表达水平升高（P〈0.05,P〈0.05）.结论 SLE患者外周血IRF5、TLR-9表达异常,可能在SLE发病中起一定作用.