石家庄地区汉族人群DIS80(pMCT118)位点VNTR基因频率分布调查

本文采用ＰＣＲ技术对石家庄地区２１６名无关个体ＤＩＳ８０（ｐＭＣＴ１１８）位点扩增片段长度多态性的等位基因频率进行调查。ＰＣＲ扩增产物采用聚丙烯酰胺凝胶电泳分型及溴化乙锭染色，共检出７０种基因型，２５个等位基因，其中以１８、２４、３０三个等位基因频率最高。将本文结果与日本人群在ＤＩＳ８０（ｐＭＣＴ１１８）位点多态性结果进行比较，石家庄地区汉族人群ＤＩＳ８０（ｐＭＣＴ１１８）位点基因频率分布符合ＨａｒｄｙＷｅｉｎｂｅｒｇ平衡定律，该位点非父排除率为０．７１，个体识别率为０．９，杂合度为０．８５。     

麻风菌基因分型及流行病学应用的研究

虽然麻风病在全世界不再是一个公共卫生问题，但是在一些国家和地区，麻风病的发现率仍旧没有明显下降。既往认为，麻风菌不能体外培养，缺乏对麻风菌的深入了解。基因组测序的成功，为麻风菌的分型提供依据。迄今为止，VNTR和SNP是麻风菌分型的基础，VNTR是由于核苷酸在复制过程中的滑动链错配引起，麻风菌的基因分型和麻风菌的地理分布和历史变迁有一定的关系，其中以VNTR为基础的地理分型和麻风病的短传播链有关，而且对于复发和再感染的鉴别具有积极意义。并探讨VNTR稳定性的意义。     

Population studies of three AMPFLPs systems in a North Polish population

Allele and phenotype frequencies for D1S80, D17S5 and ApoB were determined in a population sample of more than 200 unrelated persons from North Poland using the PCR method. For D1S80, D17S5 and ApoB 19, 13 and 21 alleles respectively were observed. No deviations from Hardy-Weinberg equilibrium were detected. All three systems have discrimination values above 92% and a cumulative discrimination index of 4.5 × 10⁴.Dedicated to Prof. Dr. J. Gerchow on the occasion of his 75th anniversary     

Establishment of Multiple Locus Variable-number Tandem Repeat Analysis Assay for Genotyping of Borrelia burgdorferi sensu lato Detected in China

Objective Human Lyme Borreliosis （LB）, which is caused by Borrefia burgdorferi sensu lato （B. burgdorferi）, has been identified as a major arthropod-borne infectious disease in China. We aimed to develop a multiple locus variable-number tandem repeat （VNTR） analysis （MLVA） assay for the genotyping of Borrelia burgdorJ：eri strains detected in China. Methods B. garinii PBi complete 904.246 kb chromosome and two plasmids （cp26 and Ip54） were screened by using Tandem Repeats Finder program for getting potential VNTR loci, the potential VNTR loci were analyzed and identified with PCR and the VNTR loci data were analyzed and MLVA clustering tree were constrcted by using the categorical coefficient and the unweighted pair-group method with arithmetic means （UPGMA）. Results We identified 5 new VNTR loci through analyzing 47 potential VNTR loci. We used the MLVA protocol to analyse 101 B. burgdorferi strains detected in China and finally identified 51 unique genotypes in 4 major clusters including B. burgdorferi sensu stricto （B.b.s.s）, B. garinii, B. a[zelii, and B. valaisiana, consistent with the current MLSA phylogeny studies. The allele numbers of VNTR-1, VNTR-2, VNTR-3, VNTR-4, and VNTR-5 were 7, 3, 9, 7, and 6. The Hunter-Gaston index （HGI） of five VNTR loci were 0.79, 0.22, 0.77, 0.71, and 0.67, respectively. The combined HGI of five VNTR loci was 0.96. Clustering of the strains of Xinjiang, Inner Mongolia and Heilongjiang was confirmed, and this situation was consistent with the close geographical distribution of those provinces. Conclusion The MLVA protocol esytablished in this study is easy and can show strains＇ phylogenetic relationships to distinguish the strains of Borrelia species. It is useful for further phylogenetic and epidemiological analyses of Borrelia strains.     

Influence of HumDN1 VNTR polymorphism on DNASE1 expression in systemic lupus erythematosus and rheumatoid arthritis

The purpose of this study was to analyze the effect of the HumDN1 VNTR polymorphism on DNASE1 mRNA expression and enzyme activity in lupus (SLE) and rheumatoid arthritis (RA) compared to healthy control (HC). Kuwait subjects (n?=?500) matched by age/gender/ethnicity were genotyped by fragment-analysis. DNASE1 expression was analysed using quantitative Real-Time-PCR and sera from subjects were screened for DNase1 reduction activity by ELISA. Allele and genotype distribution of HumDN1 VNTR revealed a significant association with susceptibility to SLE and RA (p?<?0.05, OR?>?1). Relative expression analysis revealed a significant increase in DNASE1 mRNA in SLE (p?=?0.0001) and RA (p?=?0.002) compared to HC. Stratification of subjects revealed, increased DNASE1 expression in SLE with 5/5 (p?=?0.0001), 3/4 (p?=?0.0001) and 3/5 genotype (p?=?0.01). A reduction in DNASE1 expression was specifically observed in SLE with 4,4 genotype (p?=?0.0004). RA patients with 3/4 genotype (p?=?0.02) showed a significant increase in DNASE1 expression. Similarly a significant association was observed between DNase1 reduction activity and SLE (p?=?0.0001). SLE patients with 3,4 (p?=?0.0001) and 5,5 genotype (p?=?0.0001) showed increased DNase1 reduction activity, while a lack of association was observed with RA. The present study is the first to reveal the effect of HumDN1 VNTR on DNASE1 expression in SLE and RA.     

Sex differences in circadian timing systems: Implications for disease

Virtually every eukaryotic cell has an endogenous circadian clock and a biological sex. These cell-based clocks have been conceptualized as oscillators whose phase can be reset by internal signals such as hormones, and external cues such as light. The present review highlights the inter-relationship between circadian clocks and sex differences. In mammals, the suprachiasmatic nucleus (SCN) serves as a master clock synchronizing the phase of clocks throughout the body. Gonadal steroid receptors are expressed in almost every site that receives direct SCN input. Here we review sex differences in the circadian timing system in the hypothalamic–pituitary–gonadal axis (HPG), the hypothalamic–adrenal–pituitary (HPA) axis, and sleep–arousal systems. We also point to ways in which disruption of circadian rhythms within these systems differs in the sexes and is associated with dysfunction and disease. Understanding sex differentiated circadian timing systems can lead to improved treatment strategies for these conditions.     

Endothelial Nitric Oxide Synthase Gene Intron 4 (VNTR) Polymorphism and Vascular Access Graft Thrombosis

Vascular access thrombosis is a leading cause of vascular access failure in hemodialysis patients. Thrombosis is a multifactorial condition and genetic makeup can affect thrombosis risk. We conducted a study to investigate for possible associations between ecNOS gene intron 4 variable-number tandem repeat (VNTR) polymorphism and thrombosis of polytetrafluoroethylene hemodialysis arteriovenous access grafts (AVG) in Turkish patients. Fifty-five patients with end-stage renal disease who had AVGs implanted between 2000 and 2002 and 167 healthy individuals representing our healthy population were enrolled in this prospective study. Each subject provided a venous blood sample from which DNA was isolated, and polymerase chain reaction analysis was done to identify genotypes (aa, bb, ab) for ecNOS gene intron 4 VNTR polymorphism. All grafts were placed in brachioaxillary position. The subjects were divided into two groups based on duration of graft patency. The thrombosis group (Group I) comprised 26 patients who developed AVG thrombosis in the first 12 months after placement. The no-thrombosis group (Group II) comprised 29 patients whose grafts remained patient for at least 12 months. The frequency of the aa genotype in Group I was significantly higher than that in Group II (p =. 005). At 6, 12, and 24 months, the primary patency rates for the AVGs in patients with the aa genotype were significantly lower than the corresponding rates for the bb and ab genotype groupings (p =. 01, p =. 01 and p =. 04 for the three respective time points; Kaplan–Meier). ecNOS gene intron 4 VNTR polymorphism is linked with the pathogenesis of vascular access thrombosis in Turkish patients undergoing hemodialysis.     

Congenital tuberculosis in an extremely preterm infant and prevention of nosocomial infection

Congenital tuberculosis is a rare disease, especially in non-endemic countries. We present a preterm infant who developed congenital tuberculosis in a neonatal intensive care unit (NICU). The male patient, weighing 1140 g was born by cesarean section at 26 weeks gestation. The baby's respiratory condition suddenly deteriorated at 18 days old, and he was diagnosed with congenital tuberculosis after Gram stain revealed “ghost bacilli” in his tracheal aspirate. The mother, who was born in an endemic country, had fever with unknown cause during labor and was diagnosed with miliary tuberculosis after the infant was diagnosed. Both were successfully treated for tuberculosis with a four-drug regimen. The genotyping profiles of Mycobacterium tuberculosis were identical in both mother and baby based on variable number of tandem repeat (VNTR) analysis. The lineage was considered to be East-African Indian. To prevent nosocomial infection in the NICU, 23 potentially exposed infants received isoniazid for 2 months. Two infants showed a transient liver enzyme elevation that seemed to be due to isoniazid. For 10 months after the incident, there were no infants and medical staff who developed tuberculosis. Although the incidence of tuberculosis has steadily decreased in Japan, the percentage of foreign-born individuals has increased yearly, especially those of reproductive age. The evaluation of active tuberculosis should be considered in pregnant women with unexplained fever, history of tuberculosis, or emigration from high-burden areas.     

Increased genetic diversity of the Mycobacterium tuberculosis W-Beijing genotype that predominates in eastern China

ObjectivesTo determine the genetic diversity of Mycobacterium tuberculosis (M. tuberculosis) strains in a Chinese population predominately infected with strains of the W-Beijing family.MethodsA cross-sectional study was conducted in three counties of eastern China. M. tuberculosis strains were collected at TB clinics, and patients were interviewed by trained physicians at the time of TB diagnosis. RD105 and RD181 were used to identify W-Beijing and modern W-Beijing strains, respectively, while seven-locus variable numbers of tandem repeat–mycobacterial interspersed repetitive unit (VNTR–MIRU) analysis was employed to differentiate the genotypes of these strains.ResultsOf 441 strains studied, 394 (89.3%) were identified as W-Beijing family strains; of them, 299 were modern W-Beijing strains. VNTR–MIRU identified 409 genotypes from 426 strains, including 395 unique patterns and 14 clusters. Ancestral W-Beijing strains were more likely to be clustered (OR = 1.32, 95%CI: 0.58–2.97) compared to modern W-Beijing strains. The proportions of clustered strains were 14.6%, 4.2% and 0% at sites Funing (FN), Deqing (DQ) and Yinzhou (YZ), respectively. Of the seven MIRU loci, VNTR3820 was found to have the highest discriminatory power and allelic diversity.ConclusionsVNTR–MIRU typing appears to be a reliable method for analyzing M. tuberculosis transmission in relatively closed populations. The low clustering proportions indicate that endogenous relapse may be a main source of TB cases in eastern China. Furthermore, our results indicate that migration has played may play an important role in the recent transmission of the W-Beijing family of M. tuberculosis.     

川南地区汉族人群D1S80基因座多态性分析

目的了解川南地区汉族人群DIS80位点群体遗传多态性。方法采用PCR结合聚丙烯酰胺PAGE电泳及高灵敏度银染技术,即Amp—FLP方法对川南地区120名汉族无血缘关系个体D1S80位点进行多态性分析。结果在所调查的120名无关个体样本中,观察到D1S80基因座有18个等位基因,基因频率分布在0．004166667～0．1958333333之间,杂合度为0．7869,个体识别力为0．87,非父排除率为0．8152,其基因型频率分布符合Hardy—Weinberg平衡法则。结论D1S80基因座的PCR分型具有较好的准确性和灵敏度,为法医学个体识别和亲权鉴定、遗传病基因链锁分析等的研究及应用提供了川南地区有用的遗传信息。