吴茱萸RAPD体系构建及道地性遗传背景研究

Objective To establish molecular marker method of RAPD and to probe into the genetic background of Fructus Evodia .Methods Genomic DNA of Fructus Evodia was extracted by the DNA extraction kit method and then chose the RAPD primer; RAPD Analysis on 48 individuals of Fructus Evodia from three provinces have been done. The NTSYS-PC software was used to do cluster analysis. ResultsRAPD Analysis method was optimized for Fructus Evodia. RAPD Cluster analysis revealed that variety in different kinds of Fructus Evodia was significant Evodia rutaecarpa from Liuyang and E rutaecarpa var officinalis from other places gathered in two populations at the similarity coefficient of 0.68.In the similarity coefficient of 0.78, the samples fromTongren, Liuyang, Jiande, and Wenzhou gathered in clusters, the same as their region ethnic groups, respectively.Conclusion Genetic background of Fructus Evodia from varies of regions is obviously different and RAPD analysis is a good manner to probe geo authentic crude drug Fructus Evodia.     

Intake of Volatile N-nitrosamines and Their Ability to Exogenously Synthesize in the Diet of Inhabitants from High-risk Area of Esophageal Cancer in Southern China

Objective Nan'ao County in Guandong Province is a high-risk area of esophageal cancer in Southern China. Of the suspected etiological factors in the environment, N-nitrosamines and their precursors have received the greatest attention. Methods Sixty samples of the diet ingested by the inhabitants were collected and detected for volatile N-nitrosamines and their precursors. Five N-nitrosamines detected by Gas Chromatography-Thermal Energy Analyzer were N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosopyrrolidine, N-nitrosopiperidine and N-nitrosomethyl-benzylamine. Results The average content of 5 volatile N-nitrosamines in the diet was 312.0 μg/kg (median). The daily intake of the nitrosamines was 286.5μg/head/day. Only the ability to exogenously synthesize N-nitrosopiperidine was powerful among 5 volatile N-nitrosamines. By a computerized stepwise regression analysis and curve fitting, we studied the correlation among the nitrosamines, the precursors and the major food items in the samples. Co     

Disease risks of childhood obesity in China

Objective To estimate the relative risks of dyslipidemia, hypertension, diabetes mellitus, and metabolic syndromes among overweight and obese Chinese children compared with their normal weight counterparts. Methods Overweight and obesity were defined by age- and sex-specific BMI classification reference for Chinese children and adolescents. Pediatric metabolic syndrome （MetS） and each risk factor for MetS were defined using the criteria for US adolescents. Definition of hyper-TC, LDL, and dyslipidemia for adults was applied as well. General linear model factor analysis and chi-square test were used to compare the difference in metabolic indicators among normal weight, overweight, and obese groups. Multiple logistic regression analysis was performed to estimate the odds ratio of metabolic abnormalities between obesity, overweight, and normal weight children, after adjustment for living area, family economic level, age, sex, and daily exercise time and TV watching time, as well as different dietary indices in the model. Results Significant increases in blood lipids, glucose, and blood pressure were found among overweight and obese children as compared with their counterparts with normal weight. By applying WGOC-recommended BMI classification, the risks for hypertriglyceridemia, low HDL and dyslipidemia among overweight children were 1.9, 1.4, and 1.5 times, and was 3.3, 1.5, and 1.8 times among obese groups compared to their counterparts with normal weight after adjustment for age, sex, region, socioeconomic status, physical activity, and dietary intakes. The overweight and obese children （15-17.9 years） had a high-risk of developing hypertension, which was 2.3 and 2.9 times higher than their counterparts with normal weight. Above 90% obese adolescents had abdominal obesity, while less than 1% normal weight ones had abdominal obesity. No obese adolescents were free from any risk factors for MetS, while 36.9% of normal weight adolescents were from the risk factors. 83.3% obese boys and all obese girls had metabolic syndrome, while only 15.5% normal weight boys and 18.8% normal weight girls had metabolic syndrome. Four risk factors for metabolic syndrome were found in 8.3% obese boys while none in normal weight boys and girls. The prevalence of MetS among normal weight, overweight, and obesity groups was 1.5%, 18.3%, and 38.1% respectively. Conclusion The cardiovascular disease （CVD） risk factors are clustered in obese Chinese children. Our observations strongly suggest that efforts should be made to prevent the onset of overweight and its associated diseases during early childhood.     

Serum Thyroglobulin——A Sensitive Biomarker of Iodine Nutrition Status and Affected by Thyroid Abnormalities and Disease in Adult Populations

Objective To evaluate the usefulness of the thyroglobulin(Tg) level in adults as a nutritional biomarker of iodine status and to identify the factors related to the serum Tg level. Methods A cross-sectional study was conducted in adult populations of areas differing in iodine nutrition from three provinces(Autonomous Region) in China. Serum levels of thyroid hormones and Tg as well as thyroid autoantibodies were measured. The thyroid volume and nodule were measured by ultrasound. A multivariate linear regression analysis was used to assess iodine intake and other indeterminate factors associated with the serum Tg level. Results A total of 573 adults were recruited for this study. The serum Tg levels differed significantly among the three groups(22.27 μg/L, 9.73 μg/L and 15.77 μg/L in the excess, more-than-adequate, and deficient groups, respectively). The results of multivariate linear regression analysis indicate that excess and deficient iodine intake, goiter, thyroid nodule, hypothyroidism are significantly related with higher Tg level, and Tg Ab positivity is significantly related with lower serum Tg. Conclusion The serum Tg level reflects abnormal thyroid function and is a sensitive functional biomarker of iodine nutrition status.     

Intake of Volatile N—nitrosamines and Their Ability to Exogenously Synthesize in the Diet oif Inhabitants from High—risk Area of Esophageal Cancer in Southern China

Objective Nan'ao County in Guandong Province is a high-risk area of esophageal cancer in Southern China. Of the suspected etiological factors in the environment, N-nitrosamines and their precursors have received the greatest attenfion. Methods Sixty samples of the diet ingested by the inhabitants were collected and detected for volatile N-nitrosamines and their precursors. Five Nnitrosamines detected by Gas Chromatography-Thermal Energy Analyzer were N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosopyrrolidine, N-nitrosopiperidine and N-ditrosomethyl-benzylamine. Results The average content of 5 volatile N-nitrosamines in the diet was 312.0 μg/kg (median). The daily intake of the nitrosamines was 286.5 μ tg/head/day. Only the ability to exogenously synthesize N-nitrosopiperidine was powerful among 5 volatile N-nitrosamines. By a computerized stepwise regression analysis and curve fitting, we studied the correlation among the nitrosamines, the precursors and the major food items in the samples. Conclusion It demonstrated that a relatively high content of volatile N-nitrosamines was present in the diet collected in the area.     

Genetic diversity analysis of Penicillium marneffei isolated from AIDS patients in Guangdong, China using randomly amplified polymorphic DNA

Background  Penicillium marneffei (P. marneffei) is an emerging pathogenic fungus that can cause invasive mycosis in patients with AIDS. The epidemiological features of P. marneffei infection in AIDS patients in Guangdong province remain unclear so far. This study aimed to investigate the genetic diversity within a population of 163 P. marneffei isolates obtained from AIDS patients and search for the dominant clinical strains in Guangdong province.

Methods  One hundred and sixty-three P. marneffei isolates obtained from AIDS patients in Guangdong province during January 2004 and December 2009 were studied by randomly amplified polymorphic DNA (RAPD) using two random primers (H2 and H22). The degree of similarity between samples was calculated through similarity coefficients from RAPD fragment data and the dendrogram was assessed using the unweighted pair group method with arithmetic mean (UPGMA).

Results  Two primers showed a high degree of discrimination and good stability. Primer H2 yielded eight different patterns (H2-1 to H2-8) among 163 isolates with the discriminatory power being 0.413. Primer H22 identified seven types (H22-1 to H22-7) among 163 isolates with the discriminatory power being 0.467. Genetic similarity coefficients based on RAPD data among 163 P. marneffei isolates ranged from 0.681 to 0.957, 61.96% of which were no less than 0.83. The discriminatory power of the two primers was 0.524. One hundred and sixty-three P. marneffei isolates were clustered into nine distinct groups (groups I to IX) at the similarity coefficient value of 0.83 and group I was the most common, including 101 strains (61.96%).

Conclusion  The RAPD analyses could provide important information as to the degree of genetic diversity and the relationship among clinical P. marneffei isolates, revealing genetic polymorphism and dominant strains.

     

The Expression and Change of the β3 Integrin Subunit and Fibronectin in Normal Human Oviductal Tissue and Tubal Ectopic Pregnant Tissue

Objective To investigate the expression and change of the B3 integrin subunit and fibronectin in normal human oviductal t‘issue during various phases of the menstrual cycle and tubal ectopic pregnant tissue Methods Samples of normal ( n=29 ) and pregnant fallopian tube ( n=22 ) tissues were obtained from women who had normal cycle and history of normal pregnancy. Normal oviductal tissue samples were divided into 4 groups based on their menstrual cycle. Both expression and distribution of the B3 subunit and fibronectin were determined with the immunohistochemical method and the image analysis. Results The B3 subunit was expressed in the cytoplasm of ciliated cells. The expression level of the B3 subunit was higher after ovulation than that before ovulation in isthmus epithelium (P ＜0.001), and declined significantly after ovulation in ampullae epithelium (P ＜0.001). In umbrella epithelium within 4 days after ovulation, the expression level of the B3 subunit was observed at rather higher level among other phases (P ＜0.001). The ciliated and secretory cells of the epithelium except for where the pregnancy occurred in tubal pregnancy expressed the subunit, and no significant relationship was found between the normal tubal tissue of the secretory phase and tubal ectopic pregnant tissue (P＞0.05). Fibronectin was expressed in the basementmembrane of human oviductal epithelium and matrix. The expression level of fibronectin was higher in the hyperplastic phase than that in the secretory one (P＜0.001). And it was lower in normal tubal tissue of the secretory, phase than that in tubal ectopic pregnant tissue (P ＜0.001). Conclusion The B3 integrin subunit was expressed in the ciliated cells of human oviductal epithelium, and fibronectin was expressed in the basement membrane of human oviductal epithelium and matrix. Their expression and change in oviductal tissue is based on different phases of menstrual cycle. The B3 subunit could not related to the occurrence of tubal ectopic pregnancy. Fibronectin could be the potential molecular basis for the tubal ectopic pregnancy.     

蛋白激酶B1基因多态性与重性抑郁障碍事件相关电位的关联性研究

Objective To explore the relationship of protein kinase B1 ( PKB1 ) gene polymorphisms in PI3-K pathway of BDNF and event-related potentials in depression.Methods The design of case-control research was used ,and 91 major depressive patients and 65 normal controls who were made in age and gender matched with patients were measured auditory event-related potential P300 and contingent negative variation ( CNV ) in the day when two groups were collected.Polymerase chain reaction (PCR) and direct DNA sequencing technology were used to detect PKB1 gene polymorphisms.Three SNPs that named rs3001371 ,rs2494738 ,rs1130214 were selected from 3 representative BLOCK Districts of PKB1.Two independent samples t test was used to analysis P300 and CNV between two groups,and the same way to analysis the average level of P300 and CNV and PKB1 SNP genolatency of P2(P＜0.05) and lower amplitude of P3a(P＜0.01 ) ,P3b(P＜0.01 ) and P3 (P＜0.01 ) ;CNV had der had statistical difference (P＜ 0.05 )in PKB1 rs3001371 gene between C/C and C/T genotype combined which included C allele, and T/T genotype.The amplitude of P3a( (5.93 ± 2.35 ) μV, P3b(6.51 ± 3.00) μV, P3 (6.27±2.43) μV) were lower than TT Genotype ( (7.45 ±2.19)μV, (8.63 ±3.57)μV,(8.04 ±2.57)μV,respectively).The mean of CNV indicators were not found different in statistics among the rs3001371 genotypes.Conclusions PKB1 gene rs3001371 polymorphism is associated with the principal component of P300 amplitude in patients with Major depressive disorder which suggest that genetic factors may have a certain impact on cognitive function in the patients with Major depressive disorder.     

蛋白激酶B1基因多态性与重性抑郁障碍事件相关电位的关联性研究

Objective To explore the relationship of protein kinase B1 ( PKB1 ) gene polymorphisms in PI3-K pathway of BDNF and event-related potentials in depression.Methods The design of case-control research was used ,and 91 major depressive patients and 65 normal controls who were made in age and gender matched with patients were measured auditory event-related potential P300 and contingent negative variation ( CNV ) in the day when two groups were collected.Polymerase chain reaction (PCR) and direct DNA sequencing technology were used to detect PKB1 gene polymorphisms.Three SNPs that named rs3001371 ,rs2494738 ,rs1130214 were selected from 3 representative BLOCK Districts of PKB1.Two independent samples t test was used to analysis P300 and CNV between two groups,and the same way to analysis the average level of P300 and CNV and PKB1 SNP genolatency of P2(P＜0.05) and lower amplitude of P3a(P＜0.01 ) ,P3b(P＜0.01 ) and P3 (P＜0.01 ) ;CNV had der had statistical difference (P＜ 0.05 )in PKB1 rs3001371 gene between C/C and C/T genotype combined which included C allele, and T/T genotype.The amplitude of P3a( (5.93 ± 2.35 ) μV, P3b(6.51 ± 3.00) μV, P3 (6.27±2.43) μV) were lower than TT Genotype ( (7.45 ±2.19)μV, (8.63 ±3.57)μV,(8.04 ±2.57)μV,respectively).The mean of CNV indicators were not found different in statistics among the rs3001371 genotypes.Conclusions PKB1 gene rs3001371 polymorphism is associated with the principal component of P300 amplitude in patients with Major depressive disorder which suggest that genetic factors may have a certain impact on cognitive function in the patients with Major depressive disorder.     

A study on detecting and identifying enteric pathogens with PCR

Objective To develop a rapid and definite diagnostic test of bacterial enteritis caused by pathogenic enterobacteria, the most frequent etiologic agent of infectious enteritis in the world.Methods A set of conventional PCR assays were applied to detect and identify salmonella, shigella,and E. coli O157:H7 directly from pure culture and fecal samples.The general primers of pathogenic enterobacteria were located on the uidA gene, which were found not only in E. coli nuclear acid, but also in Shigella and salmonella genes.Shigella primer was from ipaH gene whose coded invasive plasmid relative antigen existed both in plasmid and in genome.The primers of salmonella were designed from the 16SrRNA sequence.The primer of E.coli O157:H7 was taken from eaeA gene.Five random primers were selected for RAPD. The detection system included common PCR,semi-nested PCR and RAPD. Results This method was more sensitive, specific and efficient and its processing was rapid and simple. For example, the method could be used to specifically detect and identify salmonella, shigella, and E. coli O157:H7,and its sensitivity ranged from 3 to 50CFU, and its detection time was 4 hours. Conclusion This PCR method, therefore, can serve as a routine and practical protocol for detecting and identifying pathogenic microorganisms from clinical samples.     

单叶蔓荆种质资源遗传多态性的RAPD分析

目的：利用RAPD技术分析野生单叶蔓荆（Vitex trifolia L.var.simplicifolia Cham.）种质资源的遗传多态性及种内遗传差异。方法：本研究以7个不同产区的野生蔓荆为试材,采用RAPD技术对其遗传物质进行分析,结合NTSYS-2.10e软件进行遗传相似系数计算和UPGMA方法聚类分析。结果：10个随机引物共获得88条DNA谱带,其中46条（52.3%）具有多态性;采用CTAB法提取DNA时,先将试材在0.9%的蔗糖溶液中暗培养48 h,有利于提高DNA提取率和纯度。结论：各种质之间均存在一定的遗传差异,与地域关系和主要有效成分的化学差异并不一致,尚需进一步的植物逆境生理和功能基因组学研究来解释这一现象。     

微波辅助萃取牡荆挥发油的气相色谱-质谱分析

目的提供一种新型提取牡荆挥发油的方法。方法采用微波辐射法提取牡荆的挥发油，运用毛细管气相色谱-质谱联用法结合计算机检索，对其化学成分分别进行分析和鉴定，用面积归一化法测定各化合物在挥发油中的相对含量。结果共鉴定出16个化合物，挥发油的主要成分为石竹烯（20．14％）、4-羟基-苯甲酸（21．16％）、桉叶油素（12．79％）。结论通过微波辅助萃取法可以高效提取牡荆挥发油。     

单叶蔓荆子种子的最佳采收期

目的：研究单叶蔓荆子种子的最佳采收期。方法：本文以蔓荆子种子的发芽率为指标来考察单叶蔓荆子种子最佳采收期,种子的寿命和储藏方法。结果：江西省单叶蔓荆子种子的最佳采收期为每年的10-11月份,储藏方式对蔓荆子种子在一年内的发芽率无显著影响。结论：蔓荆子种子与蔓荆子药材在采收时间上存在一定的差异。     

闽产蔓荆子研究现状与展望

目的:分析福建产蔓荆子的研究现状。方法:对现有福建产蔓荆子的质量研究、生态效应、规范化种植等综合分析。结果和结论:蔓荆子是福建省的地产药材与主药材之一,应对不同部位入药进行研究,综合利用资源。     

固相微萃取/气相色谱/质谱法分析蔓荆子挥发性化学成分

目的:对蔓荆子挥发油的化学成分进行了分析。方法:采用固相微萃取/气相色谱/质谱(SPME/GC/MS)联用技术。结果:分离出81个组分,确认了其中64种组分,并用归一化法测定其相对百分含量,其相对含量占总挥发性组分峰面积的96.85%。结论:主要成分为:α-蒎烯(4.26%)、1,8-桉树脑5.90(%)、芳樟醇(3.59%)、莰烯(4.89%)、丁子香酚(2.18%)、β-石竹烯(14.72%)、β-古芸烯(2.56%)、双环吉马烯(4.35%)、α-雪松醇(11.29%)。     

栀子及其近缘类群的随机扩增多态DNA分析

目的:探索栀子与其变种雀舌栀子、重瓣栀子及变型水栀子之间的亲缘关系。方法:从56个随机引物中筛选出12个引物对栀子及其近缘类群进行随机扩增多态DNA分析,应用SPSS10.0分析软件中的Jaccard方法计算任意两样品间的相似系数,用组内聚类方法计算得出聚类树状图。结果:共得到251个有效位点。聚类分析显示,栀子与重瓣栀子最先聚类,然后与水栀子聚类,最后与雀舌栀子聚类。结论:重瓣栀子与栀子亲缘关系最近,雀舌栀子与栀子的亲缘关系最远。     

新疆药用桑树9个栽培群体的RAPD分析

目的 从DNA分子水平上分析新疆药用桑树资源9个栽培群体的遗传关系。方法 从100个随机引物中筛选出10个引物，用这10个引物做RAPD扩增，应用NTSYS软件对扩增结果进行聚类分析。结果 共扩增出108个位点。其中多态性位点91个，多态比例为84．26％，各群体间存在较明显的遗传分化。另外，获得药桑的8个特异位点。结论 RAPD分析结果与新疆药用桑树资源植物的遗传关系的传统划分是基本一致的。     

卫星搭载药用植物曼陀罗遗传变异的随机扩增多态性DNA分析

以曼陀罗种子为材料探讨太空环境对药用植物的影响。方法利用返回式卫星搭曼陀罗的种子,返回地面后播于实验田中,应用随机扩增多态性ＤＮＡ（ＲＮＡ（ＲＡＰＤ）分子标记技术分析不同组曼陀罗基因组的变异情况。结果从６５个供试引物中筛选出１５个能够产生可重复多态性增产物的引物。与地面对照组相比,失重组共产生３９条多态性片段,基因组的多态性频率为２３．１％,击中组共产生４５条多态性片段,基因组的多态性频率为２４．     

丹参主要居群的遗传关系及药材道地性的初步研究

目的 研究丹参主要居群的遗传关系及药材的道地性问题。方法 选择丹参主要产地样本44个（包括9个居群样本），进行RAPD分析，得到的扩增条带用NTSYS－pc和AMOVA软件进行数据处理。结果 （1）从100多个引物中选择出11个多态性强和重复性好的引物，扩增得到129条带；（2）不同居群内多态位点比率为20.9%－55.0%；（3）用UPGMA法得到所有样本的聚类图，可分为6个主要分支组和3个组外个体，其中四川中江居群的5个样本聚为一组，并与其他样本遗传距离较大；（4）按产地分组，遗传差异的80.44%存在于居群内，8.29%来自于组内居群间，11.27%的遗传差异来自于组间。结论 （1）丹参居群内遗传多样性十分丰富；（2）山东和河南的栽培居群栽培种源来自于当地野生居群，尚没有进行人工选择，丹参酮ⅡA等成分的减少的原因主要是栽培条件不理想；（3）地区间居群的遗传分化不均衡，四川中江和河北承德居群与其他居群距离较远；（4）丹参的道地性的确定应当依据现代的优质药材评价系统，山东和河南产的丹参也可主为是丹参的道地药材。     

云南铁线莲属12种药用植物的RAPD分析

目的:通过对12种铁线莲属药用植物进行遗传多样性和亲缘关系分析,为生药鉴定提供依据.方法:随机扩增多态性DNA(RAPD)技术分析遗传多样性,UPGMA类平均法进行聚类分析.结果:利用筛选出的10个随机引物对供试材料DNA进行随机扩增,共得到89条带,其中多态性带有89条,多态性百分率为100%.采用UPGM聚类法对扩增出的谱带进行遗传聚类分析,得出反映各种间亲缘关系的树状图.对多数种而言,种间差异明显,同种不同分布点也存在差异,但同一分布点的不同个体差异较小.结论:该标记技术对铁线莲属植物的生药鉴定是可行的,在系统学方面的应用有待进一步研究.