中国沿海地区汉族男性人群白细胞介素-1β-31C/T基因多态性与痛风的易感性研究

目的 探讨山东省沿海地区中国汉族男性群体的白细胞介素(IL)-1β启动子区rs1143627(-31C/T)基因多态性的分布状况及其与痛风易感性之间的关系.方法 选取208例痛风患者和210名健康对照者,应用聚合酶链反应限制性片段长度多态性(RFLP)技术,检测中国汉族男性群体的IL-1β启动子区rs1143627(-31C/T)位点基因多态性与痛风发病的遗传易感性的关系.采用Hardy-Weinberg检验确认标本的群体代表性,数据分析采用χ2检验和t检验.结果 痛风组中IL-1β启动子区-31C/T位点CC,CT和TT基因型分别为32.7%,43.3%和24.0%,健康对照组分别为31.9%,46.2%和21.9%,2组比较差异无统计学意义(χ2=0.427,P>0.05);2组的等位基因频率C和T间差异也无统计学意义(分别为54.3%,55.0%;45.7%,45.0%;χ2=0.038,P>0.05).经χ2检验,IL-1β基因-31C/T位点基因多态性与痛风病的危险因素无显著性关联.结论 尚不能认为中国沿海地区汉族男性人群中IL-1β启动子区rs1143627(-31C/T)基因多态性与痛风有关联性.

Abstract：

Objective To explore gene polymorphism of the C/T genotype of rs1143627 in the promoter of IL-1β gene in male population living in the coastal area of Shandong, and thus to investigate the relationship between the gene polymorphism of IL-1β and gout. Methods A total of 208 gout patients and 210 healthy controls were enrolled. The possible association between the polymorphism of IL-1 β -3 1C/T and gout in Chinese were investigated and genotype frequencies and allelic frequencies was calculated by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method. Hardy-Weinberg was used to verify the representativeness of the sample. Comparisons between the groups were performed with χ2 test and t-test. Results The frequencies of CC, CT, and TT genotypes were 32.7%, 43.3% and 24.0%,respectively among gout patients, while they were 31.9%,46.2% and 21.9%, respectively among the controls.There was no statistically difference in IL-1β -31C/T genotype frequencies between gout patients and controls (χ2=0.427, P>0.05). The allele frequencies of C and T in gout cases were different from those in the controls (54.3%, 55.0%; 45.7%, 45.0%; χ2=0.038, P>0.05). Moreover, no association between IL- I β-31 C/T genotypes and risk factors for gout were observed in gout cases by χ2 test. Conclusion Results of the present study suggest that the C/T genotype of rs1143627 in the promoter of IL-1β gene is not associated with gout in male population living in the coastal area of Shandong.     

Polymorphisms of interleukin-1B and interleukin-1 receptor antagonist genes in patients with chronic hepatitis B

AIM：To investigate the relationships between polymorphisms of interleukin-lB (IL-1B) promoter region -511CLT and interleukin-1 receptor antagonist gene (IL-1RN) and susceptibility to chronic hepatitis B in Chinese population. METHODS: Genomic DNA was extracted from the peripheral blood of 190 patients with chronic hepatitis B and 249 normal controls and then subjected to polymerase chain reaction (PCR) amplification. The PCR products were digested by restriction endonuclease AvaI. The products of digestion were subjected to 20 g/L gel electrophoresis and ethidium bromide staining. RESULTS: The frequencies of IL-1B (-511) genotypes CC, CT and TT in patients with chronic hepatitis B were 23.7%, 49.5% and 26.8%, while 26.1%, 47.4% and 26.5% respectively in controls. The results showed that there was no significant difference in the frequencies of alleles or genotypes in IL-1B between patients with chronic hepatitis B and controls. The distributions of IL-1B (-511) genotype CC were significantly different between the two subgroups (HBV-DNA ≤1×10^3 copies/mL as subgroup I, HBV-DNA> 1×10^3 copies/mL as subgroup Ⅱ) of chronic hepatitis B (P=0.029). Only four of the five kinds of polymorphism (1/1, 1/2, 2/2 and 1/4) were found in this study. The frequencies of IL-1RN genotypes 1/1, 1/2, 2/2 and 1/4 were 88.9%, 9.0%, 0.5% and 1.6% in patients with chronic hepatitis B respectively, while were 81.1%, 16.9%, 0.4% and 1.6% respectively in controls. The frequencies of genotype1/2 and IL-1RN allele 2 in patients with chronic hepatitis B were lower than those in controls (P=0.016 and P=0.029, respectively). CONCLUSION: There is an association between polyrnorphisms of the promoter region -511C/T of IL-1B and IL-1RN intron 2 and chronic hepatitis B virus infection. SubJects with IL-1RN 2 may be resistant to HBV infection, and IL-1B(-511) geNotype CC is closely related with HBV-DNA replication, which gives some new clues to the study of pathogenesis of chronic hepatitis B.     

Polymorphisms of interleukin-1B and interleukin-1 receptor antagonist genes in patients with chronic hepatitis B

AIM:To investigate the relationships between polymorphismsof interleukin-1B(IL-1B)promoter region -511C/T andinterleukin-1 receptor antagonist gene(IL-1RN)andsusceptibility to chronic hepatitis B in Chinese population.METHODS:Genomic DNA was extracted from the peripheralblood of 190 patients with chronic hepatitis B and 249 normalcontrols and then subjected to polymerase chain reaction(PCR)amplification.The PCR products were digested byrestriction endonuclease AvaI.The products of digestion weresubjected to 20 g/L gel electrophoresis and ethidium bromidestaining.RESULTS:The frequencies of IL-1B(-511)genotypes CC,CT and Tr in patients with chronic hepatitis B were 23.7%,49.5% and 26.8%,while 26.1%,47.4% and 26.5%respectively in controls.The results showed that there wasno significant difference in the frequencies of alleles orgenotypes in IL-1B between patients with chronic hepatitisB and controls.The distributions of IL-1B(-511)genotypeCC were significantly different between the two subgroups(HBV-DNA≤1×10~3 copies/mL as subgroup I,HBV-DNA>1×10~3 copies/mL as subgroup Ⅱ)of chronic hepatitis B(P=0.029).Only four of the five kinds of polymorphism(1/1,1/2,2/2 and 1/4)were found in this study.The frequenciesof IL-1RN genotypes 1/1,1/2,2/2 and 1/4 were 88.9%,9.0%,0.5% and 1.6% in patients with chronic hepatitis B respect,while were 81.1%,16.9%,0.4% and 1.6% respectively incontrols.The frequencies of genotypel/2 and IL-1RN allele2 in patients with chronic hepatitis B were lower than thosein controls(P=0.016 and P=0.029,respectively).CONCLUSION:There is an association between polymorphismsof the promoter region -511C/T of IL-1B and IL-1RN intron 2and chronic hepatitis B virus infection.Subjects with IL-1RN2 may be resistant to HBV infection,and IL-1B(-511)genotype CC is closely related with HBV-DNA replication,which gives some new clues to the study of pathogenesis ofchronic hepatitis B.     

Association of polymorphisms of interleukin-18 gene promoter region with chronic hepatitis B in Chinese Han population

AIM: To investigate the polymorphisms of interleukin-18 (IL-18) gene promoters, and to disclose whether such polymorphisms are associated with susceptibility to chronic hepatitis B in Chinese Han population. METHODS: Using polymerase chain reaction with sequence specific primers (PCR-SSP) method, the single nucleotide polymorphisms (SNPs) of the promoter region of IL-18 gene at position -607 and -137 were detected in 231 patients with chronic hepatitis B and 300 normal controls. RESULTS: Allele C at position -607 in the promoter of IL-18 gene was detected in 48.7% of normal controls and 51.9% of patients, while allele A at position -607 was detected in 51.3% of normal controls and 48.1% of patients. The frequencies of -607CC, -607 CA and -607AA genotypes in normal controls were 22.0%, 53.3% and 24.7% respectively and in chronic hepatitis B patients were 26.8%, 50.2% and 23.0% respectively. Allele G at position -137 in the promoter of IL-18 gene was detected in 82.3% of normal controls and 88.5% of chronic hepatitis B patients, while allele C at position -137 was detected in 17.7% of normal controls and 11.5% of patients. The frequencies of -137GG, GC and CC genotype were 67.3%, 30.0% and 2.7% in normal controls respectively, while in chronic hepatitis B patients were 78.8%, 19.5% and 1.7% respectively. The frequency of-137GG genotype in chronic hepatitis B groups was significantly higher than that in normal controls (x2=8.55, P=0.003 <0.05), whereas the frequencies of -607C/-137C and -607A/-137C haplotypes in chronic hepatitis B groups were significantly lower than that in normal controls. The association between genotypes of IL-18 promoter region polymorphisms and HBV copies showed that the frequency of -607AA genotype in high HBV-DNA copies groups was lower than that in low HBV-DNA copies groups (x2=6.03, P=0.014 <0.05). CONCLUSION: The polymorphisms of the promoter region of IL-18 gene at position -607 and -137 are closely associated with susceptibility to chronic hepatitis B. The people with allele C at position -137 in the promoter of IL-18 gene may be protected against HBV infection; moreover AA genotype at position -607 may be closely linked to inhibit HBV-DNA replication. These findings give some new clues to the study of pathogenesis of chronic hepatitis B.     

E-cadherin gene C-160A promoter polymorphism and risk of non-cardia gastric cancer in a Chinese population

AIM: To test the hypothesis that E-cadherin gene (CDH1) C-160A promoter variant genotype is associated with an increased risk for developing gastric cancer. METHODS: In this population-based case-control study of gastric cancer in Jiangsu Province, China, we performed polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to genotype the C-160A polymorphism of CDH1 promoter in 206 non-cardia gastric cancer patients and 261 age- and sex-matched but unrelated cancer-free controls. RESULTS: The frequencies of genotypes CC, CA and AA were 57.8%, 36.4% and 5.8% in gasfric cancer cases, respectively, and 58.2%, 34.9% and 6.9% in controls respectively. The distributions of CDH1 genotypes were not significantly different between gastric cancer cases and controls (P=0.87 for genotype frequency and P=0.92 for allele frequency). Compared with the CC genotype, the CA and AA genotypes were not associated with an increased risk for non-cardia gastric cancer (adjusted odds ratios (OR)=1.15, and 95% confidence interval (95% CI)=0.78-1.72 for CA genotype, and OR = 0.90 and 95% CI = 0.42-2.01 for AA genotype). CONCLUSION: E-cadherin gene C-160A promoter polymorphism may not play a major role in the etiology of non-cardia gastric cancer in Chinese population.     

Relationship between interleukin 18 polymorphisms and susceptibility to chronic hepatitis B virus infection

AIM:To identify the relationship between the tagging single nucleotide polymorphism sites(tagSNPs)of the Interleukin-18(IL-18)gene and genetic susceptibility to chronic hepatitis B virus infection in Chinese patients.METHODS:Five hundred and one cases of chronic hep-atitis B virus(HBV)infection and 301 HBV natural clearance controls were studied.Two tagSNPs in the IL-18 gene(rs1946518A/C and rs574424C/G)were genotyped by the Multiplex Snapshot technique.The genotype and allele frequencies were calculated and analyzed.RESULTS:In the genotypes of rs1946518,the AA type was present at a higher frequency in the patients compared to those in the controls.Odds ratio(OR)of theAA genotype for the comparison with that of the AC and the CC genotype was 1.537(95%confidence intervals(CI):1.116-2.218,P=0.009<0.025).In pheno-types,the allele C at rs1946518 was of a significantly lower frequency in the patients with chronic hepatitis B than that in the controls(P=0.017<0.025).OR of the allele A for the comparison with that of the allele C was 1.279(95%CI:1.045-1.567).As for the rs574424 genotypes,no significant difference in this genotype distribution or in this allele frequency between the patients and the control subjects was observed.No significant difference in the haplotype frequencies between the patients with chronic hepatitis B and HBV natural clearance individuals was displayed.CONCLUSION:The data suggest that genotype AA and the allele A of the IL-18 at position rs1946518 are closely associated with the resistance to chronic hepatitis B and may be the dangerous gene.However,no statistical association was found between polymorphisms of rs574424 for IL-18 and hepatitis B.     

Interleukin-10 -1082 promoter polymorphism is not associated with susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma in a population of high-incidence region of north China

AIM: To investigate the possible association of G→A single nucleotide polymorphism (SNP) at the -1082 position of interleukin (IL)-10 promoter with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA) in a population of a high incidence region of North China. METHODS: IL-10-G1082A promoter SNP was genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) in 355 cancer patients (203 ESCC and 152 GCA) and 443 healthy controls. RESULTS: Smoking significantly increased the risk of ESCC and GCA development (the age and sex adjusted OR = 1.42 and 2.64, 95%CI = 1.11-1.81 and 1.46-4.76, respectively). Similarly, family history of upper gastrointestinal cancer (UGIC) significantly increased the risk of developing ESCC and GCA (the age and sex adjusted OR = 1.44 and 3.10, 95%CI = 1.18-1.75 and 1.94-4.97, respectively). The A/A, A/G and G/G genotype frequencies of IL-10-G1082A were 60.3%, 37.0% and 2.7% in healthy controls, 57.6%, 39.9% and 2.5% in ESCC and 61.2%, 36.8% and 2.0% in GCA patients, respectively. The frequencies of A and G alleles were 78.8% and 21.2% in healthy controls, 77.6% and 22.4% in ESCC patients and 79.6%, 20.4% in GCA patients. The distribution of genotype and allelotype in ESCC and GCA patients was not significantly different from that in healthy controls (P>0.05). Compared to the A/A genotype, the combination of A/G and G/G genotypes did not show a significant effect on the risk of developing ESCC and GCA; the adjusted odds ratio was 0.92 (95% CI = 0.76-1.11) in ESCC and 0.95 (95% CI = 0.61-1.46) in GCA, respectively. When stratified for smoking status and family history of UGIC, the combination of A/G and G/G genotypes also did not show any significant influence on the risk of ESCC and GCA development compared to A/A genotypes. CONCLUSION: IL-10-G1082A polymorphism might not be used as a stratification marker to predicate the risk of ESCC and GCA development in North China.     

Effect of cytokine gene polymorphism on histological activity index, viral load and response to treatment in patients with chronic hepatitis C genotype 3

AIM: To investigate the association between cytokine gene polymorphism and disease status in chronic hepatitis C genotype 3 by liver biopsy, ALT, HCV RNA levels and response to treatment. METHODS: Patients with chronic hepatitis C genotype 3 were analyzed for single nucleotide polymorphisms of interleukin (IL)-10, IL-1 beta, interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-β) by polymerase chain reaction using sequence-specific oligonucleotide primers. Liver biopsies were assessed by modified histological activity index (HAI) scoring system using a scale of 0-18 for grading the necro-inflammatory activity and 0-6 for staging the fibrosis. HCV RNA levels were determined by bDNA assay. The patients were treated with interferon alpha and ribavirin for 6 mo. Sustained virological response was assessed 6 mo after the completion of the treatment. RESULTS: Out of the 40 patients analyzed, 26 were males. Mean age was 40.5±12.5 years (range 18-65 years). The frequencies of different dimorphic polymorphisms based on single nucleotide substitution were as follows: IL-10-1082 G/A 85%, A/A 12.5%, G/ G 2.5%; IL-10-819 A/C 87.5%, C/C 10%, A/A 2.5%; IL-10-592 C/A 72.5%, C/C 27.5%; IL-1 C 90%, U 10%; IFN-874 T/A 50%, T/T 27.5%, A/A 22.5%; TNF-308 A/G 95%, GIG 5%; TGF-10 T/C 52.5%, C/C 35%, T/T 12.5%. The mean grades of necro-inflammatory activity of different genotypes of IL-10 at promoter site -1082 were A/A = 3.6, A/G = 5.0, and G/G = 10.0 and the difference was significant (P = 0.029). The difference in the stage of disease at a scale of 0-6 was A/A 0.8, A/G 2.3, and G/G 4.0 (P = 0.079). The difference in the HAI seemed to be related to the presence of allele -1082G. For IL-10 -819 genotypes, mean scores of fibrosis were A/A = 6.0, A/C = 2.2, and C/C = 1.0 (P = 0.020) though the inflammatory activity was not much different. No significant differences in HAI were noted among polymorphisms of other cytokines. Moreover, ALT and HCV RNA levels were not significantly different among different cvtokine polymorphisms. There was a significant correlation of HAI and HCV RNA levels with the duration of disease. TGFBBB -10 genotype CC patients had a better end of treatment response than those with other genotypes (P = 0.020). Sustained virological response to the treatment was not influenced by the cytokine polymorphism. No effect of other factors like viral load, degree of fibrosis, gender, steatosis, was observed on sustained virological response in this population infected with genotype 3. CONCLUSION: There is no significant correlation between cytokine polymorphisms and HAI except for the polymorphisms of anti-inflammatory cytokine IL-10, which may influence hepatic inflammatory activity and fibrosis in patients with chronic hepatitis C genotype 3. Sustained virological response in this genotype does not seem to be influenced by cytokine gene polymorphisms.     

Apolipoprotein AI gene polymorphisms and risk for coronary artery disease in Chinese Xinjiang Uygur and Han population

Objective To analyze the relationship between polymorphism at the Apolipoprotein AI (Apo AI) gene and the risk for coronary artery disease. Methods A total of 107 patients (mean age 56 ±11 years) diagnosed as having stable angina pectoris (SAP) (23 cases), unstable angina pectoris (UAP) (23 cases) or myocardial infarction (MI) (61 cases) were prospectively evaluated.DNA was obtained from the 107 patients and 50 controls. In order to determine the Apo AI genotypes at two polymorphic sites (G/A at -75 bp, and C/T at+83 bp), DNA was PCR amplified and digested with MspI. Results The frequency of carriers of the rare allele at the - 75 bp site (Ml-) was 0.49 in cases and 0.30 in controls ( P< 0. 05). The frequencies of the Ml - allele among patients with SAP, UAP, MI and controls were 0. 37 (vs. controls, P > 0. 05), 0.54 (vs. controls, P < 0.05), 0.52 (vs. controls, P< 0. 05) and 0. 30, respectively. The frequencies for carriers of the rare allele at the + 83bp polymorphism (M2) were observed among patients     

Association of TNFA(-308G/A), IFNG(+874 A/T) and IL-10(-819 C/T) polymorphisms with protection and susceptibility to dengue in Brazilian population

Objective: To evaluate gene polymorphisms and their association with susceptibility to dengue.Methods: A retrospective case-control study was performed with 262 subjects,comprising 78 dengue fever(DF) patients, 49 dengue hemorrhagic fever(DHF) patients and 135 healthy controls. Genotypic and allelic profiles were identified using polymerase chain reaction based in real time and amplification-refractory mutation system.Results: We observed a protective association of IL-10(-819 C/T) C allele(P = 0.028,OR = 0.56, CI = 0.34–0.91) against DHF, while the C/T(P = 0.047, OR = 2.10,CI = 1.01–4.38) and T/T(P = 0.008, OR = 3.82, CI = 1.38–10.59) genotypes were associated with DHF and DF, respectively. The dominant model TNFA-308 GA + AA(P = 0.043, OR = 0.45, CI = 0.20–1.00) genotypes were found to have protective effect against dengue infection. A protective association among the IFNG(+874 A/T) A/T genotype against DF(P = 0.02, OR = 0.46, CI = 0.24–0.89) and DHF(P = 0.034,OR = 0.43, CI = 0.19–0.95) was observed. When the studied single-nucleotide polymorphism was analyzed in combination, the combination GTA(P = 0.022, OR = 2.95,CI = 1.18–7.41) was statistically significantly associated with susceptibility to DF and the combination GCT(P = 0.035, OR = 0.28, CI = 0.08–0.90) with protection against the development of DHF.Conclusions: This research identifies the association of the IFNG(+874 A/T), TNFA(-308 G/A), IL-10(-819 C/T) genotypes as a factor for protection, susceptibility and severity to dengue.     

Distribution and effects of polymorphic RANTES gene alleles in HIV/HCV coinfection -- a prospective cross-sectional study

AIM: Chemokines and their receptors are crucial for immune responses in HCV and HIV infection. RANTES gene polymorphisms lead to altered gene expression and influence the natural course of HIV infection. Therefore, these mutations may also affect the course of HIV/HCV coinfection. METHODS: We determined allele frequencies of RANTES-403 (G→A), RANTES-28 (C→G) and RANTES-IN1.1 (T→C) polymorphisms using real-time PCR and hybridization probes in patients with HIV (n = 85), HCV (n = 112), HIV/HCV coinfection (n = 121), and 109 healthy controls. Furthermore, HIV and HCV loads as well as CD4+ and CD8+ cell counts were compared between different RANTES genotypes. RESULTS: Frequencies of RANTES-403 A, RANTES-28 G and RANTES-IN1.1 C alleles were higher in HIV infected patients than in healthy controls (-403: 28.2% vs 15.1%, P = 0.002; -28: 5.4% vs 2.8%, not significant; IN1.1: 19.0% vs 11.0%, P = 0.038). In HIV/HCV coinfected patients, these RANTES alleles were less frequent than in patients with HIV infection alone (15.4% P = 0.002; 1.7%; P = 0.048; 12.0%; not significant). Frequencies of these alleles were not significantly different between HIV/HCV positive patients, HCV positive patients and healthy controls. CONCLUSION: All three RANTES polymorphisms showed increased frequencies of the variant allele exclusively in patients with HIV monoinfection. The finding that the frequencies of these alleles remained unaltered in HIV/HCV coinfected patients suggests that HCV coinfection interferes with selection processes associated with these alleles in HIV infection.     

IL-10 and TNF-α promoter haplotypes are associated with childhood Crohn＇s disease location

AIM： To determine the distribution and frequencies of the genotypes and haplotypes of the genes encoding for the glucocorticoid receptor （GR）, the tumor necrosis factor （TNF）-α and the interleukin （IL）-10 in childhood Crohn＇s disease （CD） and to assess the impact of the corresponding DNA variants on clinical and disease phenotypes.
METHODS： Ten variants in GR, TNF-a and IL-10 were genotyped in 113 childhood CD cases and 95 healthy subjects, both of French-Canadian origin.
RESULTS： For the GR polymorphisms （R23K and N363S） and IL-10 variants in the 5＇flanking region （-1082 G 〉 A, -819 T 〉 C and -592 A 〉 C）, no difference was observed in allele and genotype frequencies between CD patients and controls. At the haplotype level, we found three IL-10 haplotypes previously described in Caucasians （GCC, ACC and ATA） and three novel haplotypes only present in IBD patients. When we analyzed the haplotype distribution with the anatomical location of the disease, the GCC haplotype was associated with the colonic and the ACC haplotype with the terminal ileum location, respectively. The genotyping of five polymorphisms in the promoter region of the TNF-α gene （-1031 T 〉 C, -863 A 〉 C, -857 T 〉 C, -308 A 〉 G and -238 A 〉 G） revealed a significant overrepresentation of homozygous -1031 CC among CD patients （OR = 9.9） and an association with the colonic location. For TNF-α, eleven haplotypes were inferred, including two frequent ones, TCCGG and CACGG, which were significantly observed more frequently in controls and cases, respectively.
CONCLUSION： This is one of the first studies investigating the association between haplotype structure and disease location in a CD pediatric cohort. Our results will help to increase our understanding of the genetic determinants of childhood CD.     

运用高分辨率熔解曲线技术检测痛风患者SLC22A12基因intron5(+4668C/T)多态性

Objective To investigate a new single nucleotid polymorphism (SNP) intron5(+4668C/T) in SLC22A12 in primary gout patients and the association between clinical characteristics and genotypes. Methods One hundred and one primary gout patients and 186 healthy subjects were recruited into this study. Blood pressure, body mass index (BMI) was recorded. Serum uric acid, glucose, lipid and creatinine were detected. DNA was extracted from peripheral blood to amplify the fragment located in intron 5. The genotypes of SLC22A12 can be detected with high-resolution melting (HRM) assay, followed by sequencing analysis. Chi-square test was used for statistical analysis. Results ① A new SNP in intron 5 of SLC22A12 was identi-fied successfully by HRM, which was defined as intron 5 (+4668C/T). CC, CT and TT genotypes were unam-biguously distinguished with HRM technology, which was fully concordant with sequencing. ②The genotypes of CC, CT and TT in male and female groups were 28.1%, 33.7%, 38.2% and 20.0%, 47.1%, 32.9%, respectively.③ However, no significant differences of genotype distribution were found concerning BMI, blood pressure, creatinine, total cholesterol and triglyceride in both male group and female group. But the serum uric acid levels in the CC genotype were significantly higher than those with the CT+TT genotypes. ④ The genotype frequencies of CC and CT+TT in high uric acid group were remarkably different from those in low uric acid group (21.2%, 78.8%,; 35.0%, 65.0%; P<0.05). Conclusion A new SNP has been successfully discovered with HRM technology with simplicity, rapidity and accuracy. T allele of intron 5 (+4668C/T) may be a genetic protective factor for hyperuricemia among Chinese population.     

运用高分辨率熔解曲线技术检测痛风患者SLC22A12基因intron5(+4668C/T)多态性

Objective To investigate a new single nucleotid polymorphism (SNP) intron5(+4668C/T) in SLC22A12 in primary gout patients and the association between clinical characteristics and genotypes. Methods One hundred and one primary gout patients and 186 healthy subjects were recruited into this study. Blood pressure, body mass index (BMI) was recorded. Serum uric acid, glucose, lipid and creatinine were detected. DNA was extracted from peripheral blood to amplify the fragment located in intron 5. The genotypes of SLC22A12 can be detected with high-resolution melting (HRM) assay, followed by sequencing analysis. Chi-square test was used for statistical analysis. Results ① A new SNP in intron 5 of SLC22A12 was identi-fied successfully by HRM, which was defined as intron 5 (+4668C/T). CC, CT and TT genotypes were unam-biguously distinguished with HRM technology, which was fully concordant with sequencing. ②The genotypes of CC, CT and TT in male and female groups were 28.1%, 33.7%, 38.2% and 20.0%, 47.1%, 32.9%, respectively.③ However, no significant differences of genotype distribution were found concerning BMI, blood pressure, creatinine, total cholesterol and triglyceride in both male group and female group. But the serum uric acid levels in the CC genotype were significantly higher than those with the CT+TT genotypes. ④ The genotype frequencies of CC and CT+TT in high uric acid group were remarkably different from those in low uric acid group (21.2%, 78.8%,; 35.0%, 65.0%; P<0.05). Conclusion A new SNP has been successfully discovered with HRM technology with simplicity, rapidity and accuracy. T allele of intron 5 (+4668C/T) may be a genetic protective factor for hyperuricemia among Chinese population.     

Genetic association of cytokines polymorphisms with autoimmune hepatitis and primary biliary cirrhosis in the Chinese

AIM: To characterize gene polymorphism of several cytokine gene in-patients with AIH and PBC and to analyze the difference of the polymorphism distribution between Chinese patients and healthy controls. METHODS: The study population consisted of 62 patients with AIH, and 77 patients with PBC. The genetic profile of four cytokines was analyzed by restriction fragment length polymorphism after specific PCR amplification (PCR-RFLP) or sequence-specific primers PCR (SSP-PCR). The analyzed gene polymorphism included interleukin-1 (IL-1) (at position +3 953 and IL-1RN intron 2), IL-6 (at position -174), IL-10 promoter (at position -1082, -819, and -592). The control group consisted of 160 healthy blood donors. RESULTS: The majority of Chinese people including patients and healthy controls exhibited IL-1B 1,1 genotype, and there was no significant difference in AIH, PBC patients and controls. There were highly statistically significant differences in the distribution of the IL-1RN gene polymorphism between the patients with PBC compared with controls. The frequency of IL-1RN 1,1 was significantly higher (90.9% vs 79.4%, P=0.03) and the frequency of IL-1RN 1,2 was significantly lower in PBC patients (6.5% vs 17.5%, P=0.01). No statistical difference was observed between AIH patients and controls. All of the 160 healthy controls and 62 cases of AIH patients exhibited IL-6-174GG genotype, and there were four cases, which expressed IL-6-174GC genotype in 77 cases of PBC patients. The frequency of IL-6-174GC was markedly significantly higher in PBC patients compared with controls (5.2% vs 0%, P=0.004). No statistically significant difference was found in the distribution of IL-10 promoter genotype in AIH and PBC patients compared with controls. CONCLUSION: The polymorphisms of IL-1RN and IL-6-174G/C appear to be associated with PBC in Chinese patients.     

Relationship between β-fibrinogen gene - 455G/A,- 148C/T polymorphisms and atrial fibrillation with cerebral infarction

Objective To investigate the relationship between β-fibrinogen (β-Fg) gene -455G/A,- 148C/T polymorphisms and atrial fibrillation (AF) with cerebral infarction.Methods Polymerase chain reaction-restriction fragment length polymorphism was used to detect the genotypes of -455G/A and -148C/T in 97 patients with AF with cerebral infarction (AF with cerebral infarction group),80 patients with AF alone (AF group),and 98 health subjects (healthy control group).Results The A allele frequencies of -455G/A in the AF with cerebral infarction group and the AF group were 0.304 and 0.344 respectively.They were significantly higher than 0.179 in the healthy control group (all P＜0.05).The T allele frequencies of - 148C/T in the AF with cerebral infarction group and the AF group were 0.348and 0.369,respectively.They were significant higher than 0.240 in the healthy control group (all P＜0.05).Conclusions The A allele of -455G/A and the T allele of -148C/T were associated with AF with cerebral infarction and AF.     

Cytochrome P450 2E1 genetic polymorphism and gastric cancer in Changle, Fujian Province

AIM:Genetic polymorphism in enzymes of carcinogenmetabolism has been found to have the influence on thesusceptibility to cancer.Cytochrome P450 2E1(CYP2 E1)isconsidered to play an important role in the metabolicactivation of procarcinogens such as N-nitrosoamines andlow molecular weight organic compounds.The purpose ofthis study is to determine whether CYP450 2E1polymorphisms are associated with risks of gastric cancer.METHODS:We conducted a population based case-controlstudy in Changle county,Fujian Province,a high-riskregion of gastric cancer in China.Ninety-one incidentgastric cancer patients and ninety-four healthy controls wereincluded in our study.Datas including demographiccharacteristcs,diet intake,and alcohol and tobaccoconsumption of indivduais in our study were completad by astandardized questionnaire.PCR-RFLP revealed threegenotypes:heterozyote(C1/C2)and two homozygotes(C1/C1 and C2/C2)in CYP2E1.RESULTS:The frequency of variant genotypes(C1/C2 andC2/C2)in gastric cancer cases and controls was 36.3% and24.5%,respectively.The rare homozygous C2/C2 genotypewas found in 6 indivduals in gastric cancer group(6.6%),whereas there was only one in the control group(1.1%).However,there was no statistically significan differencebetween,the two groups(two-tailed Rsher's exact test,P=0.066).Indlvduals in gastric cancer group were more likelyto carry genotype C1/C2(odds ratio,OR=1.50)and C2/C2(OR=7.34)than indlvduals in control group(x~2=4.597,fortrend P=0.032).The frequencies of genotypes with the C2allele(C1/C2 and C2/C2 genotypes)were compared withthose of genotypes without C2 alleie(C1/C1 genotype)among indlvduals in gastric cancer group and control groupaccording to the pattem of gastric cancer risk factors.Theresults show that indlvduals who exposed to these gastriccancer risk factors and carry the C2 allele seemed to have ahigher risk of developing gastric cancer.CONCLUSION:Polymorphism of CYP2E1 gene may havesome effct in the development of gastric cancer in Changle county,Fujian Province.     

GSTM1, GSTT1, GSTP1 and CYPIA1 genetic polymorphisms and susceptibility to esophageal cancer in a French population： Different pattern of squamous cell carcinoma and adenocarcinoma

AIM: To evaluate the association between CYPIA1 and GSTs genetic polymorphisms and susceptibility to esophageal squamous cell carcinoma (SCC) and esophageal adenocarcinorna (ADC) in a high risk area of northwest of France.METHODS: A case-control study was conducted to investigate the genetic polymorphisms of these enzymes (CYPIAI*2C and GSTP1 exon 7 Val alleles, GSTM1 *2/*2 and GSl-l-l*2/*2 null genotypes). A total of 79 esophagealcancer cases and 130 controls were recruited. RESULTS: GSTM2*2/*2 and CYP1A1*1A/*2C genotype frequencies were higher among squamous cell cardnomas at a level close to statistical significance (OR = 1.83, 95% CI0.88-3.83, P= 0.11; OR = 3.03, 95% CI 0.93-9.90, P= 0.07,respectively). For GSTP1 polymorphism, no difference wasfound between controls and cases, whatever their histological status. Lower frequency of GST/-1 deletion was observed in ADC group compared to controls with a statistically significant difference (OR=13.31, 95% CI 1.66-106.92, P&lt;0.01).CONCLUSION: In SCC, our results are consistent with the strong association of this kind of turnout with tobacco exposure. In ADC, our results suggest 3 distinct hypotheses:(1) activation of exogenous procarcinogens, such as small halogenated compounds by GSTTI‘, (2) contribution of GSTT1 to the inflammatory response of esophageal mucosa, which is known to be a strong risk factor for ADC,possibly through leukotriene synthesis; (3) higher sensitivity to the inflammatory process associated with intracellular depletion of glutathione.     

Frequencies of poor metabolizers of cytochrome P450 2C19 in esophagus cancer,stomach cancer,lung cancer and bladder cancer in Chinese population

AIM:To investigate the association between cytochromeP450 2C19 (CYP2C19) gene polymorphism and cancersusceptibility by genotyping of CYP2C19 poor metabolizers(PNs) in cancer patients.METHODS:One hundred and thirty-five cases of esophaguscancer,148 cases of stomach cancer,212 cases of lungcancer,112 cases of bladder cancer and 372 controls weregenotyped by allele specific amplification-polymerase chainreaction (ASA-PCR) for CYP2C19 PMs.The frequencies ofPMs in cancer groups and control group were compared.RESULTS:The frequencies of PMs of CYP2Clg were 34.1%(46/135) in the group of esophagus cancer patients,31.8%(47/148) in the stomach cancer patients,34.4%(73/212) inthe group of lung cancer patients,only 4.5%(5/112) in thebladder cancer patients and 14.0%(52/372) in control group.There were statistical differences between the cancer groupsand control group (esophagus cancer,x~2=25.65,P<0.005,OR=3.18,95%CI=2.005-5.042;stomach cancer,x~2=21.70,P<0.005,OR=2.86,95% CI=1.820-4.501;lung cancer,x~2=33.58,P<0.005,OR=3.23,95% CI=1.503-6.906;bladdercancer,x~2=7.50,P<0.01,OR=0.288,95% CI=0.112-0.740).CONCLUSION:CYP2C19 PMs have a high incidence ofesophagus cancer,stomach cancer and lung cancer,converselythey have a low incidence of bladder cancer.It suggests thatCYP2C19 may partidpate in the activation of procarcinogen ofesophagus cancer,stomach cancer and lung cancer,but mayinvolve in the detoxification of carcinogens of bladder cancer.     

过氧化氢酶基因-262C/T及-21A/T多态性与燃煤型氟中毒相关性的研究

目的 探讨过氧化氢酶(CAT)基因启动子区-262C/T、-21A/T多态性与燃煤型氟中毒之间的关系.方法 2007年,在贵州省毕节市鸭池镇燃煤型氟中毒非改灶村抽取150名村民作为非干预组,于该市长春镇燃煤型氟中毒改灶村抽取150名村民作为干预组,于非地方性氟中毒长顺县白云山镇抽取150名村民作对照组,应用聚合酶链反应限制性片段长度多态性(PCR-RFLP)分析方法,分别检测各组CAT-262C/T、-21A/T多态的基因型.结果 非干预组、干预组和对照组CAT-262C/T、CAT-21A/T多态的基因型频率分布符合Hardy-weinberg遗传平衡定律(P＞0.05).在研究人群中检测到CAT-262C/T多态的两种基因型CC和CT,未检测到TT基因型;检测到CAT-21A/T多态的三种基因型AA、AT和TT.对照组、干预组和非干预组CAT基因-262C/T多态位点的基因型(CC、CT)频率分别为89.33%(134/150)、10.67%(16/150)、88.67%(133/150)、11.33%(17/150)、9333%(140/150)、6.67%(10/150);对照组、干预组和非干预组C等位基因频率分别为94.67%(284/300)、94.33%(283/300)、96.67%(290/300);T等位基因频率分别为5.33%(16/300)、5.67%(17/300)、3.33%(10/300).对照组,干预组和非干预组CAT基因-21A/T多态位点基因型(AA、AT、TT)频率分别为48.67%(73/150)、46.00%(69/150)、5.33%(8/150)、52.67%(79/150)、38.00%(57/150)、9.33%(14/150)、51.33%(77/150)、38.00%(57/150)、10.67%(16/150);对照组、干预组和非干预组A等位基因频率分别为71.67%(215/300)、71.67%(215/300)、70.33%(2ll/300);T等位基因频率分别为28.33%(85/300)、28.33%(85/300)、29.67%(89/300).CAT-262C/T和CAT-21A/T基因型频率及等位基因频率在对照组,干预组和非干预组中的分布,组间比较差异均无统计学意义(x2=0.331、0.336,P均＞0.05).结论 CAT基因-262C/T、-21A/T多态性与燃煤型氟中毒无明显的相关性.

Abstract：

Objective To explore the relationship between -262C/T and -21A/T polymorphisms of catalase(CAT) gene and coal-burning borne fluorosis. Methods In 2007, 150 villagers were taken as a nonintervention group in Bijie city from the village of coal-burning borne fluorosis areas with unchanged cooking stoves;150 villagers were taken as the intervention group from the town of Changchun county where cooking stoves changed; 150 villagers were taken as control from non-endemic fluorosis areas in Baiyun town of Changshun county.PCR-restriction fragment length polymorphism were employed to detect genotypes of CAT-262C/T and CAT-21A/T polymorphism of CAT gene. Results The genotypic frequencies of CAT-262C/T and CAT-21A/T in nonintervention group,intervention group and control group were in line with Hardy-Weinberg equilibrium law (P＞ 0.05 ).The genotypes of CC and CT were detected while no TT were detected for CAT-262C/T polymorphism; the genotypes of AA, AT and TT were detected for CAT-21A/T. The genotype frequencies of CAT-262 CC, CT in control group, intervention group and non-intervention group were (89.33%(134/150), 10.67%(16/150); 88.67%(133/150), 11.33% (17/150),93.33% (140/150),6.67% (10/150), respectively. The gene frequency of C in control group, intervention group and non-intervention group were (94.67% (284/300), 94.33% (283/300),96.67%(290/300), respectively. The gene frequency of T in control group, intervention group and non-intervention group were 5.33%(16/300), 5.67%(17/300), 3.33%(10/300), respectively. The genotype frequencies of CAT-21 AA,AT and TT in control group, intervention group and non-intervention group were 48.67%(73/150),46.00%(69/150),5.33%(8/150) ,52.67%(79/150) ,38.00%(57/150) ,9.33% (14/150) ,51.33%(77/150) ,38.00%(57/150), 10.67%(16/150), respectively. The gene frequency of A in control group, intervention group and non-intervention group were 71.67%(215/300),71.67%(215/300),70.33%(211/300), respectively. The gene frequency of T in control group, intervention group and non-intervention group were 28.33% (85/300),28.33% (85/300),29.67% (89/300),respectively. CAT-262C/T and CAT-21A/T genotype and allele frequencies in the control group, the intervention group and non-intervention group showed no significant differences in the distribution(x2= 0.331,0.336, all P ＞0.05 ). Conclusion CAT-262C/T and CAT-21A/T polymorphism is not associated with coal-burning borne fluorosis.