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.     

新疆哈萨克及维吾尔族SLE和RA患者MBL基因多态性调查

目的 对新疆哈萨克及维吾尔族人群中系统性红斑狼疮(SLE)及类风湿关节炎(RA)患者甘露糖结合凝集素基因54位密码子点突变进行测定并分析.方法 PCR-RFLP.结果 42例哈萨克族SLE患者基因频率:0.536(GGC),0.464(GAC);24例维吾尔族SLE患者基因频率:0.667(GGC),0.333(GAC);22例哈萨克族RA患者基因频率:0.614(GGC),0.386(GAC);15例维吾尔族RA患者基因频率:0.733(GGC),0.267(GAC).结论 MBL不同的基因型和这些疾病发生可能存在相关.     

多浪羊和中国美利奴羊MHC-DRB1基因多态性与包虫病的遗传易感性

目的探讨多浪羊和中国美利奴(新疆军垦型)羊MHC-DRB1基因的多态性与包虫病的遗传易感性。方法利用巢式PCR扩增192只多浪羊和199只中国美利奴(新疆军垦型)羊(其中各有70只和98只为棘球蚴病感染——包虫病阳性)MHC-DRB1基因第二外显子,其296bp扩增产物经SacI、Hin1I和HaeIII3种限制性内切酶进行RFLP多态性分析。结果多浪羊和中国美利奴羊的MHC-DRB1基因第二外显子在SacI、Hin1I和HaeIII酶切位点存在丰富的多态性,两绵羊品种均检测出了2、2和6种等位基因,但基因型分别为3、3、18和3、3、15种。DRB1第二外显子的第159,173,177,178,208,225位碱基上存在多态性。结论将两绵羊品种包虫病阴性和阳性羊的等位基因频率和基因型频率分别进行了分析比较,结果提示多浪羊等位(基因HaeIII a与包虫病的易感性相关(P<0.05);基因型SacI ab和HaeIII cf与包虫病的抗性相关(P<0.05),SacI bb,Hin1I ab和HaeIII aa 3种基因型与包虫病的易感性相关(P<0.05),而基因型HaeIIIbe和HaeIII ef与包虫病的易感性具有较强的相关性(P<0.01)。中国美利奴(新疆军垦型)羊等位基因HaeIII d与包虫病的易感性相关(P<0.05);基因型SacI ab和HaeIII ee与包虫病的抗性相关(P<0.05),而基因型HaeIII bd与包虫病的易感性相关(P<0.05)。     

云贵两省三地带绦虫的分子鉴定——核糖体DNA第一内转录间隔区（ITS1）限制性酶切片段长度多态性（RFLP）分析

目的用PCR-RFLP方法对rDNA-ITS1片段进行分析,以进一步明确云贵地区是否存在牛带绦虫亚洲亚种,并建立一种快速鉴定方法. 方法 取贵州都匀株（DY）、贵州从江株（CJ）、云南大理株（DL）带绦虫及台湾株（TW）成虫标本,剪取孕节,抽提DNA,PCR扩增rDNA- ITS1片段,分别用4种限制性内切酶MspI、CfoI、AluI、RsaI对扩增片段作酶切分析. 结果 PCR产物经AluI、RsaI酶切后, TW、DL、DY和CJ株RFLP图谱一致;经MspI、CfoI酶切后,TW、DL和DY株RFLP图谱一致,CJ株显著不同. 结论 1）DL和DY株与TW株同属牛带绦虫亚洲亚种;而CJ株是传统牛带绦虫;2）rDNA-ITS1的PCR-RFLP分析方法简便,可以用于带绦虫的分类学研究.     

云贵两省三地带绦虫的分子鉴定--核糖体DNA第一内转录间隔区(ITS1)限制性酶切片段长度多态性(RFLP)分析

目的用PCR-RFLP方法对rDNA-ITS1片段进行分析,以进一步明确云贵地区是否存在牛带绦虫亚洲亚种,并建立一种快速鉴定方法. 方法 取贵州都匀株(DY)、贵州从江株(CJ)、云南大理株(DL)带绦虫及台湾株(TW)成虫标本,剪取孕节,抽提DNA,PCR扩增rDNA- ITS1片段,分别用4种限制性内切酶MspI、CfoI、AluI、RsaI对扩增片段作酶切分析. 结果 PCR产物经AluI、RsaI酶切后, TW、DL、DY和CJ株RFLP图谱一致;经MspI、CfoI酶切后,TW、DL和DY株RFLP图谱一致,CJ株显著不同. 结论 1)DL和DY株与TW株同属牛带绦虫亚洲亚种;而CJ株是传统牛带绦虫;2)rDNA-ITS1的PCR-RFLP分析方法简便,可以用于带绦虫的分类学研究.     

福建省部分地区乙型肝炎病毒基因型分布及其临床意义的探讨

目的:首次调查福建省五个地区乙型肝炎病毒(HBV)基因型的分布状况,并探讨HBV基因型与HBV,相关肝病临床的可能相关性。方法:收集福州市、厦门市、泉州市、三明市、莆田市等地区慢性HBV感染者的血清,采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法检测HBV基因型,应用多分类logistic回归分析、对应分析研究HBV基因型与临床的相关性。结果:431份HBV DNA阳性的血清中基因B型275例(63.8％),C型100例(23.2％),D型及其混合型共51例(11.8％)。未见A、E、F型。多分类logistic回归分析显示泉州和三明地区HBV基因B型所占比例显著高于福州地区(P=0.002;P=0.006);无症状携带者、慢性肝炎、重型肝炎都是以基因B型为主要基因型;基因C型在肝硬化中所占比例(47％)显著高于无症状携带者(14.5％)和重型肝炎组(14.7％)(P=0.009,P<0.001);基因B型的e抗原阳性率(52.4％)显著低于C型(56％)(P=0.008);基因D型患者e抗原阳性率(30.8％)也低于C型(P=0.051)。对应分析表明HCC与基因D型及其混合型关系密切。结论:①福建省HBV感染以基因B型为主,其次是C型,也存在基因D型的流行。②福建省部分地区基因型B和C的分布可能存在差异。③基因B型在年轻患者中可能与重型肝炎的发展有关;基因C型在年长患者中可能更易导致肝硬化。④基因D型与     

Incidence of Giardia lamblia Subspecies by PCR-RFLP in Stool Specimens of Hospitalized Children at Urmia Mutahhari Hospital,West Azerbaijan Province,Iran

Background

Giardia lamblia is one of the most prevalent intestinal flagellate protozoa that infects a wide range of vertebrate hosts causing severe intestinal disorder in children.This study was performed to determine subspecies of G.lamblia by the PCR-RFLP method, targeting the glutamate dehydrogenase(gdh)locus, in hospitalized children at Urmia Mutahhari Hospital, West Azerbaijan Province,Iran and determining the infection transformational storages in this area.

Methods

Overall, 720 stool specimens were collected from the hospitalized children, 34 samples were positive and Giardia cysts were detected under the microscope. Cysts were partially purified by the sucrose density gradient method and then washed with sterile distilled water to remove effectively the PCR inhibitors. Genomic DNA of G. lamblia isolates was extracted by freeze-thaw cycles followed by phenol/ chloroform/isoamyl alcohol method. The single step PCR-RFLP assay was used to differentiate the assemblages between A and B, which were found in humans. In this method, 432 bp expected size was amplified, and then for detection of subspecies, specific restriction RsaI and BspLI enzymes were used.

Results

Totally 34 samples were positive in terms of Giardia cyst out of 720 examined samples microscopically, so the parasite spread rate is reported 4.72%. Analysis PCR-RFLP on these samples revealed that 28 samples (93.3%) have the genotype BIII and 2 samples (6.7%) belong to the subgroup BIV.

Conclusion

PCR-RFLP is a proper analytical method for determining the genotype among parasite types, using the glutamate dehydrogenizes zone’s genes. Based on the results, an animal origin of infection cycle is suggested.     

Molecular and Parasitological Study of Cryptosporidium Isolates From Cattle in Ilam,West of Iran

Background

Cryptosporidiosis is one of the most important parasitic infections in human and animals. This study was designed for survey on the prevalence of Cryptosporidium infection in farms of Ilam, west of Iran, using parasitology method and genotyping by Nested PCR-RFLP.

Methods

Fecal samples of 217 cattle were collected fresh and directly from the rectum of cattle. All of the samples were examined by microscopic observation after staining with modified Ziehl-Neelsen (MZN). Genomic DNA extracted by using EURx DNA kit. A Nested PCR-RFLP protocol amplifying 825 bp fragment of 18s rRNA gene conducted to differentiate species and genotyping of the isolates using SspI and VspI as restriction enzymes.

Results

The prevalence of Cryptosporidium infection in cattle using both methods is 3.68%. Most of the positive cattle were calves under six months. Species diagnosis carried out by digesting the secondary PCR product with SspI that C. parvum generated 3 visible bands of 448, 247 and 106 bp and digested by VspI restriction enzyme generated 2 visible bands of 628 and 104bp. In this investigation all of the positive samples were Cryptosporidium parvum.

Conclusion

C. parvum (bovine genotype) detected in all positive cattle samples in Ilam, west of Iran. The results of the present study can help for public health care systems to prevention and management of cryptosporidiosis in cattle and the assessment of cattle cryptosporidiosis as a reservoir for the human infection.