Interleukin 6 −174G>C polymorphism and cancer risk: Meta-analysis reveals a site dependent differential influence in Ancestral North Indians

In our earlier studies, single nucleotide polymorphisms (SNPs) associated with anti-inflammatory cytokines were found to influence risk for breast cancer in western Indian women. Analysis of Interleukin 6 (IL-6) −174G>C polymorphism in this cohort (patients = 182; controls = 236) suggested a protective role for IL-6 −174C allele associated with the lower expression of the cytokine (OR = 0.54; 95% CI 0.32–0.89, dominant model). Together these observations suggested that in comparison to Caucasians, inflammation associated-cytokine gene polymorphisms may have higher influence on risk for cancer in this population. To examine this possibility we analyzed data assessing influence of Interleukin 6 (IL-6) −174G>C polymorphism on risk for various cancers. Overall, there was a marginally higher risk for rare allele homozygotes compared to wild type homozygotes (OR = 1.07; 95% CI 1.00–1.15). Increased risks for genitourinary cancers and for skin cancer were also indicated. The ethnicity based analysis indicated a protective effect of the minor allele in Ancestral North Indians (OR = 0.73; 95% CI 0.55–0.97). Site by ethnicity analysis once again revealed a significant protection against breast cancer (OR = 0.51; 95% CI = 0.37–0.70; dominant model) but an opposite influence on the risk of genitourinary malignancies (OR = 2.51; 95% CI 1.59–3.96; recessive model) in this population alone. The observations imply that contribution of IL-6 to inflammation or effector immunity may depend on the site of malignancy. Assessment of available data in relation to prognosis in breast cancer patients also revealed trends that are compatible with the observations of the meta-analysis. Thus, IL-6 −174G>C polymorphism clearly represents a potential modulator of risk for malignant disorders with ethnicity and site dependent trends. The results also support the possibility of higher influence of inflammation related cytokine gene polymorphisms on the risk for cancers in Ancestral North Indians.     

Development of PCR based assays for detection of lethal Holstein haplotype 1, 3 and 4 in Holstein Friesian cattle

Holstein haplotype (HH) 1, 3 and 4 are lethal mutations, responsible for early embryonic losses in Holstein Friesian (HF) cattle, worldwide. Three PCR based assays – tetra Amplification Refractory Mutation System PCR, PCR primer induced restriction analysis and PCR-restriction fragment length polymorphism techniques for screening of HH1, 3 and 4, respectively were developed and validated. During screening, six among 60 HF bulls were found as carrier for either of three mutations. These PCR assays are highly accurate and reproducible and can be used for screening of the haplotypes in HF cattle.     

基于ITS序列PCR-RFLP鉴别大蓟药材及饮片中掺伪飞廉、魁蓟的方法研究

[目的]利用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）,建立快速鉴别大蓟混伪品飞廉和魁蓟的方法。[方法]通过比对ITS基因序列,分别筛选飞廉、魁蓟的限制性内切酶位点并设计鉴别引物。考察PCR反应的退火温度、循环数及不同酶的适用性,对酶切反应时间和酶切底物量进行优化。同时对该方法适应性及掺伪比例的专属性、稳定性进行考察。[结果]当退火温度为58～60℃、循环数为30个时,样品均能扩增出一条379 bp的DNA条带。底物为8μL、酶切温度为37℃、酶切反应120 min时,ZraI酶将飞廉切割成120 bp和259 bp两条DNA条带;DNA底物为8μL、酶切温度为37℃、酶切反应60 min时,ApaLI酶将魁蓟切割成126 bp和253 bp两条DNA条带。[结论]试验建立的PCR-RFLP方法能够准确地鉴别大蓟混伪品飞廉、魁蓟,避免此类药材的混用,以保证临床用药安全。     

Association of PON1-L55M Genetic Variation and Breast Cancer Risk: A Case-Control Trial

Background: Paraoxonase 1 (PON1), a multifactorial antioxidant enzyme, has a defensive role against oxidative stress, which is believed to contribute to cancer development. This study aimed to investigate the association of PON1-L55M functional polymorphism with breast cancer risk. Material and methods: In the experimental study, blood samples were collected from 150 healthy women controls and 150 breast cancer subjects. The L55M genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism. Results: Our analysis showed that the genotypes distribution is in Hardy-Weinberg equilibrium for both case and control groups. Our data revealed that there are significant associations between PON1-L55M polymorphism and breast cancer risk in homozygote (OR= 2.13, 95%CI= 1.14-4.00, p= 0.018), dominant (OR= 1.72, 95%CI= 1.07-2.76, p= 0.024), and allelic (OR= 1.55, 95%CI= 1.12-2.15, p= 0.008) models. Conclusions: Our results suggest that the PON1-L55M genetic variation could be a genetic risk factor for breast cancer risk and it could be considered as a molecular biomarker for screening of susceptible women.     

中国和泰国人群DNA修复基因XRCC1多态性研究

目的 获得DNA修复基因X线修复交叉互补基因1ARG399Gln基因型频率及基因频率在中国汉族及泰国人群中的分布特点。方法 采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法结合核苷酸序列分析检测150例中国汉族及140例泰国人群XRCClArg399Gln等位基因及基因型分布频率。结果 基因型Arg/Arg,Arg／Gln,Gln／Gln在中国汉族人群中的频率分别为50．00％,44．67％,5．30％;在泰国人群中的频率分别为62．86％,31．43％,5．71％,基因型频率分布符合Hardy—Weinberg平衡定律,中国汉族人群基因型频率分布和泰国、日本、韩国、瑞典人群相近（P〉0．05）,与美国白人、美国黑人、葡萄牙、印度人群差异有显著性（P〈0．05）。结论 XRCCl Arg399Gln基因型在中国汉族和泰国人群中分布差异无显著性,该基因多态性存在种族和地域差异。     

PCR-RFLP检测Apo CⅢ基因T-455C多态性的反应条件优化

目的：探讨聚合酶链反应-限制性片段长度多态性（PCR—RFLP）反应体系中各种成分的优化,建立检测载脂蛋白CⅢ（ApoCⅢ）T-455C多态性的方法。方法：采用PCR—RFLP检测ApoCⅢ基因T-455C多态性,同时通过对参与反应体系的成分,在一定范围内设置不同的浓度或参数组,进行正交试验和单因素逐项试验,观测各成分在不同条件下对结果的影响。结果：ApoCⅢ基因检测到3种基因型。PCR反应体系中不同模板含量、引物浓度、TaqDNA聚合酶用量、dNTP浓度、Mg^2＋浓度、退火温度和酶切体系中PCR产物量、内切酶量、酶切时间等对反应结果均有不同程度的影响。结论：该研究建立的PCR—RFLP体系检测Apo CⅢ基因T-455C多态性技术,是一种高效快速、简单敏感、准确可靠的分析方法,适合常规实验室开展。     

Polymorphism of CYP1A1 gene variants rs4646903 and rs1048943 relation to the incidence of cervical cancer in Chhattisgarh

Cytochrome P450 CYP1A1 is a phase 1 xenobiotic metabolizing enzyme involved in the metabolism of toxins, endogenous hormones and pharmaceutical drugs. It is therefore possible that polymorphism of CYP1A1 gene producing functional changes in the enzyme may be susceptible factors in cervical carcinogenesis. This study was aimed to look association of CYP1A1 m1 (T > C) and m2 (A > G) gene polymorphisms in Chhattisgarh population. In this case-control study, we analyzed leukocyte DNA from a total of 200 subjects form Chhattisgarh (100 cases and 100 controls). All subjects were genotyped for CYP1A1 m1 (T > C) and m2 (A > G) using PCR-RFLP with statistical analysis by using SPSS version 16.0 and VassarStats (online). Among the two gene variants rs4646903 (T > C) and rs1048943 (A > G), individuals with AG and GG genotypes of CYP1A1 m2 polymorphism have significantly higher and increased risk of cervical cancer (OR = 2.0, 95%CI = 1.04-3.84, p = 0.035; OR = 62.9, 95%CI = 3.72-1063.83, p = 0.004 respectively) and the association of CYP1A1 m1 polymorphism did not show any significant relationship with cervical cancer patients (p = 0.23). The ‘G’ allele showed strong association with the disease (p < 0.0001). Thus, CYP1A1 m2 polymorphism showed an increased risk in the population leading to cervical cancer. Our study suggested that the presence of ‘C’ allele of rs4646903 (T > C) showed no risk and ‘G’ allele of rs1048943 (A > G) might be a leading allele to cause increased cervical cancer susceptibility due to significant association of CYP1A1 m2 gene polymorphism.     

绞股蓝属植物及其混淆品乌蔹莓的PCR-RFLP鉴别研究

目的：寻找简便、可重复的分子标记方法对绞股蓝属Gynostemma植物及其混淆品乌蔹莓Cayratia japonica进行鉴别。方法：对7种常见药用绞股蓝属植物及其混淆品乌蔹莓的6个cpDNA片段进行PCR 扩增，再利用TaqⅠ，HpaⅡ，EcoRⅠ，RsaⅠ，HhaⅠ，HindⅢ等6种限制性内切酶分别对扩增片段进行消化。结果：36种DNA片段/内切酶组合中，trnK1f -trnK2r片段与RsaⅠ组合可将乌蔹莓从绞股蓝属植物中区分出来，产物清晰、结果稳定。结论：PCR-RFLP分析方法可有效区分常见绞股蓝属植物与其混淆品乌蔹莓。