β细胞素基因多态性与2型糖尿病患者血脂异常相关性研究

目的:探讨β细胞素基因外显子1TGC19GGC、5′非翻译区A-226G两个位点基因多态性与2型糖尿病患者血脂代谢的相关关系。方法:对130例2型糖尿病患者和96例健康对照者分别采用RFLP-PCR、SASP-PCR对两个位点进行基因多态性筛查。结果:①外显子1TGC19GGC研究中,2型糖尿病组总胆固醇、高密度脂蛋白、低密度脂蛋白、甘油三酯在两种基因型之间无显著性差异(P>0.05);健康对照组总胆固醇、甘油三酯、低密度脂蛋白在两种基因型之间无显著性差异(P>0.05),高密度脂蛋白在两种基因型之间虽无显著性差异(P>0.05),但是在TG基因型中有降低的趋势。②5′非翻译区A-226G研究中,2型糖尿病组总胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白在两种基因型之间无显著性差异(P>0.05);对照组总胆固醇、高密度脂蛋白、低密度脂蛋白两种基因型间无显著性差异(P>0.05),甘油三酯在GG基因型中明显升高(P<0.05)。结论:①BTC基因外显子1TGC19GGC的研究中发现,在2型糖尿病组中该位点基因突变与血脂水平无明显相关性,在健康对照组中TG基因型与高密度脂蛋白水平降低有一定的相关性。②BTC基因5′非翻译区A-226G的研究中发现,在2型糖尿病组中该位点基因突变与血脂水平无明显相关性,在健康对照组中GG基因型与甘油三酯水平升高有一定相关性。

西北地区汉族原发性高血压人群β_2肾上腺素受体Gln27Glu基因多态性研究

目的:研究西北地区汉族人群β2肾上腺素受体Gln27Glu基因多态性与原发性高血压的相关性。方法:选择182例西北地区汉族高血压患者及104例无高血压者,应用直接测序法分析β2肾上腺素受体Gln27Glu基因型。结果:高血压组基因型分布频率Gln/Gln 82.4%、Gln/Glu 17.6%,未发现变异性纯合子Glu/Glu;而相应对照组分别是81.7%、16.3%、1.9%,两组间基因型频率比较差异无统计学意义。高血压组Gln等位基因频率91.2%、Glu等位基因频率8.8%,而相应对照组分别是90.4%、9.6%,两组间等位基因频率比较差异无统计学意义。说明Glu等位基因可能与高血压无关。应用方差分析进行组间比较,发现三种基因型间收缩压、舒张压等指标比较差异均无统计学意义,但左心室后壁厚度、室间隔厚度和左室质量比较差异有统计学意义,Gln/Glu最高。结论:β2肾上腺素受体Gln27Glu基因多态性可能与西北地区汉族人群原发性高血压无关。

急性冠脉综合征CYP2C19基因多态性与氯吡格雷抵抗的相关性研究

目的:探讨CYP2C19基因多态性对急性冠脉综合征患者服用氯吡格雷产生抵抗的相关性。方法:对96例患者术后服用氯吡格雷,提取全血DNA,采用TaqMan探针法扩增CYP2C19多态性位点和流式细胞仪检测血小板反应性指数PRI,以PRI≥50%定义为氯吡格雷抵抗(CR)。观察CYP2C19基因型与临床危险因素对CR及预后不良事件的影响。结果:22.9%的患者发生CR。弱代谢型组比例为11.45%,CR率最高。CYP2C19*2基因型与2型糖尿病是CR的独立危险因素。弱代谢组发生心脏不良事件的频率最高。结论:CR与CYP2C19*2基因型和2型糖尿病密切相关,为判断急性冠脉综合征患者预后提供一定的临床依据。

宁夏回族人群HLA-DQB1基因多态性与类风湿性关节炎的相关性

目的:探讨宁夏回族人群HLA-DQB1基因多态性与类风湿性关节炎(RA)的相关性。方法:采用序列特异性引物聚合酶链式反应(SSP-PCR)的方法对宁夏回族66例健康个体及70例RA患者的HLA-DQB1位点进行基因分型,应用遗传学统计方法对DQB1基因多态性与RA的相关性进行分析。结果:对照组与病例组等位基因的构成及基因型分布具有统计学意义。在对照组中,0501/0501和0501/0601这2种基因型频率最高,均为10.61%,在病例组中,0301/0303基因型频率最高(17.14%),0301/0302基因型频率次之(10.00%);*0501等位基因频率在对照组中高达24.24%,而在患者中仅为5%;*0301等位基因在RA患者中频率高达28.57%,而在对照组中仅为3.79%。结论:0301/0303和0301/0302这2种基因型可能与宁夏回族RA的易患性有关,*0301等位基因可能是宁夏回族易患RA的危险因子,而*0501等位基因可能为宁夏回族抗RA的基因。

瘦素受体基因多态性与胃癌易感性研究

目的:探讨瘦素受体基因多态性与胃癌易感性的关系。方法:随机选取89例胃癌患者作为研究对象,97例健康体检者作为正常对照,采用PCR-RFLP技术检测瘦素受体基因rs1137100和rs1137101位点的多态性,同时运用ELISA法测定血清瘦素水平。结果:胃癌组瘦素受体基因rs1137100位点的AA基因型和A等位基因明显降低罹患胃癌的风险(P=0.036,OR=0.259,95%CI=0.068-0.987;P=0.008,OR=0.493,95%CI=0.291-0.833)。rs1137101位点AA和GA基因型与胃癌发病风险无明显相关性。但是,位点rs1137101的A等位基因可能会降低胃癌的发病几率(P=0.006,OR=0.446,95%CI=0.248-0.802)。胃癌组血清瘦素水平明显高于正常对照组(P<0.05)。结论:瘦素受体基因多态性与胃癌易感性密切相关,瘦素受体基因rs1137100和rs1137101的A等位基因可能会降低胃癌发病风险。

从易感基因多态性探讨原发性高血压病中医证候实质的思考

认为基因背景可能是证候形成的主要原因之一,基因的这种细微差异决定其生物行为出现极大的变化。运用基因组技术寻找不同中医证型中相关基因表达谱的差异表达,对研究中医证型的实质及指导临床用药有重大的意义。

冠状动脉痉挛患者自主神经功能相关基因多态性研究

目的:探讨冠状动脉痉挛(CAS)患者在双重特异性A型激酶锚定蛋白2功能多态性[AKAP10(A/G)I646V)]的基因型分布情况与自主神经功能的关系。方法:研究组选择冠脉造影证实的CAS患者58例,对照组选择冠脉造影阴性患者58例,采用心率变异性时域指标评价自主神经功能,比较两组患者AKAP10的基因分布情况及心率变异性指标的异同。结果:CAS组患者平均心率高于对照组(P<0.05);CAS组时域指标SDNN、SDANN、RMSSD、PNN50均明显低于对照组(P<0.05);CAS组AA、AG、GG基因型频率分别为31.0%、39.7%、29.3%,对照组则为53.4%、32.8%、13.8%。CAS组A、G等位基因的频率分别为54%、46%,对照组A、G等位基因的频率分别为70%、30%,两组患者基因型及等位基因频率分布均存在统计学差异(P<0.05)。结论:AKAP10基因多态性可能通过影响自主神经的功能而成为影响CAS的发生。

F5及PAI-1基因多态性联合检测在烧伤多次手术患者深静脉血栓形成风险评估中的应用

目的:探究F5及PAI-1基因多态性联合检测在烧伤多次手术患者深静脉血栓形成风险评估中的应用价值。方法:选取200例拟行多次手术烧伤患者为研究对象。检测F5基因表型(GG、GA、AA)及PAI-1基因表型(4G4G、4G5G、5G5G),随访患病后2个月内深静脉血栓事件发生情况。结果:本研究纳入200例患者中F5基因分布:AA 57例,GA 99例,GG患者44例;PAI-1基因分布为:4G4G74例,4G5G80例,5G5G46例。139例患者先后发现不同部位不同程度深静脉血栓。F5基因表型间血栓形成差异不具有统计学意义(P>0.05),PAI-1基因表型间血栓形成差异具有统计学意义(P<0.001)。与GG/5G5G患者相比,AA/4G4G表型患者血栓发生风险升高50倍(95%CI 2.559~976.97),AA/4G5G表型患者血栓发生风险升高20倍(95%CI 1.796~222.777),GA/4G4G表型患者血栓发生风险升高32倍(95%CI 3.068~333.798),GG/4G4G表型患者血栓发生风险升高27.5倍(95%CI 2.503~302.174),血栓事件发生率差异具有统计学意义(P<0.05)。结论:F5及PAI-1基因多态性对预测多次手术后血栓形风险具有重要意义,单独检测F5及PAI-1基因多态性存在一定局限,建议临床联合检测。

Association of Polymorphisms of Leptin Gene with Body Weight and Body Sizes Indexes in Chinese Indigenous Cattle

PCR-SSCP was used to analyze the polymorphism of leptin gene in 539 samples of six cattle breeds, namely Nanyang （NY）, Qinchuan （QC）, Jiaxianred （JXR）, Xizhen （XZ）, Luxi （LX）, and Holstein cow （HOL） breeds. PCR products with a 330 bp were amplified and sequenced. The results showed that the frequencies of alleles A/B of NY, QC, JXR, XZ, LX, and HOL breeds were 0.558/0.442, 0.492/0.508, 0.571/0.429, 0.658/0.342, 0.591/0.409, and 0.615/0.385, respectively. The association of variations of leptin gene with growth traits in NY, QC, JXR breeds was analyzed. Some indexes of the individuals with genotype BB were higher than that with genotype AA and AB in NY breed, such as the indexes of body length, heart length, body weight, hucklebone width, body height, and average day gain. The height at hip cross of the individuals with genotype BB was higher than that of those with genotype AA and AB in QC breed （P 〈 0.05）. So leptin gene may be one of the candidate genes for growth traits with height at hip cross, but not for body weight, heart length, and body length trait. However, the height at hip cross and hucklebone width of the individuals with genotype AB and BB were higher than that of those with genotype AA in JXR breed （P 〈 0.05）, but the difference was not statistically significant in body weight and body sizes （body height, body length, and heart length）. And the polymorphisms in leptin gene were caused by G→T transversion at the 66th bp position, A→C transversion at the 67th bp position and G→T transversion at the 299th bp position. These results may be applied to marker-assisted selection of Chinese cattle breeds.

RAPD analysis of genetic diversity of nine strains of Auricularia auricular cultivated in Heilongjiang Province

Polymorphism of nine strains （CF05, CF09, 29, 916, AU9, Chang10, Chang7, 8808 and AU. Japanese） of A. auricular cultivated in Heilongjiang Province were analyzed by RAPD （Random Amplication polymorphic DNA）. Thirteen primers were selected from forty PCR primers with 10bp long random primer. The results showed that nine strains of A. auricular have a high level of genetic diversity and the percentage of DNA polymorphic was 96.05. The genotypes of 9 strains of Auricularia auricular were identified by the fingerprints from primer 27 and primer 46 by RAPD analysis. The results are helpful for quickly identifying strains of A. auricular in its early breeding time, and also provides a powerful theoretic basis to differentiate strains （Auricularia auricular） whose morphology is very similar in breeding programs of edible fungus.

Study on Population Genetic Characteristics of Qinchuan Cows Using Microsatellite Markers

To evaluate the genetic polymorphisms and to search for available molecular markers for Qinchuan cattle, 90 Qinchuan cows were genotyped with 12 microsatellite markers. A total of 247 alleles were detected, with the number of alleles ranging from 13 （INRA005） to 33 （HEL13）, giving a mean number of 21 alleles per locus. The total and mean effective allele number were 142.6229 and 11.8852, respectively. Mean sampling variance of the allele frequency was 2.6036 × 10^-4. Allele size ranges of the 12 microsatellite loci were different. The observed heterozygosity and expected heterozygosity were from 0.7842 （INRA005） to 0.9775 （BM315） and 0.7952 （BM315） to 0.9446 （HEL13）, respectively. Mean observed heterozygosity and mean expected heterozygosity were 0.9117 and 0.9047, respectively. Polymorphism information content values were from 0.7653 （INRA005） to 0.9420 （HEL13）, and mean polymorphism information content of the 12 microsatellite loci was 0.8965. All the 12 microsatellite loci were highly polymorphic, which showed that there were rich genetic polymorphisms at these detected microsatellite loci in Qinchuan cows. At the 12 microsatellite loci, the mean fixation index was -0.0076, reflecting that the degree of heterozygote defect at these loci was not high and deviations from Hardy-Weinberg equilibrium were not significant.

AFLP Variation Analysis on the Germplasm Resources of Leymus chinensis

The fingerprints of 27 Leymus chinensis (Trin.) Tzvel accessions from different regions in China were generated and their genomic DNA variations were assayed via amplified fragment length polymorphism (AFLP). A total of 537 DNA fragments were amplified ranging from 75-530 bp using 8 AFLP primer combinations, among which 89 (16.6%) were monomorphic and 329 (61.3%) were polymorphic. The average number of DNA band produced by each primer combination was 66.13 and the total polymorphic rate was 78.84%. PICs (polymorphic information contents) scores for AFLP ranged from 0.0-0.5 and the mean PIC was 0.216. PIC scores were maximum (0.5) for 8.5% of the AFLP markers. All the results revealed that there was rich polymorphism in genomic DNA of L. chinensis . A DNA molecular dendrogram was established for 27 L. chinensis genotypes based on genetic variation and UPGMA cluster analysis of 537 AFLP markers amplified by 8 primer combination, suggesting that the genetic variation with high frequency showed close correlation with their ecotopes and geographical distributions. Twenty_seven L. chinensis genotypes could be divided into four groups. Among them the lower genetic relationship was shown. The reason of high genetic variation frequency in L. chinensis species and formation of varieties was discussed.

RAPD and ISSR Markers of Fertility Restoring Gene for Aegilops kotschyi Cytoplasmic Male Sterility in Wheat

LK783 was found to be a good fertility restorer for K-type male sterility of wheat (Triticum aestivum L.). RAPD and ISSR (inter-simple sequence repeat polymorphism) markers were employed to map the major restoring gene in LK783. Maintainer and restorer DNA pools were established using the extreme sterile and fertile plants among KJ5418A//911289/LK783 F 1 population, respectively. Four hundred and eighteen RAPD primers and 33 ISSR primers were used for screening polymorphisms between the two pools, and amplification bands using a RAPD primer of OPK18 and an ISSR primer of UBC-845 were found polymorphic between the two pools. Linkage analysis showed that OPK18 450 and UBC-845 800 were linked to the restoring gene in LK783. The distance between the restoring gene and OPK18 450 was (15.07±6.28) cM (centiMorgan), with the distance between the restoring gene and UBC-845 800 being (8.20±4.85) cM. The marker of UBC-845 800 was located on chromosome 1BS by amplifying nulli-tetrasomics and 1B ditelosomics of Chinese Spring with the primer of UBC-845, indicating that the restoring gene in LK783 was located on 1BS. The breeding for new fertility restorer lines of K-type cytoplasmic male sterility of wheat would be facilitated by using the two markers.

Isolation and Characterization of Microsatellite DNA Markers from Forest Musk Deer(Moschus berezovskii)

In this report, we describe the isolation and characterization of microsatellite loci for forest musk deer （Moschus berezovskii） through an improved enrichment protocol. Five new markers were isolated from the genomic DNA of forest musk deer and showed high polymorphism with 4 - 13 alleles in 24 sampled individuals from the population of Jinfeng Mountain, Sichuan Province, China. The observed and expected heterozygesities were from 0.429 - 0.957 and 0.587 - 0.902, respectively. The average polymorphism information content （PIC） value in these five loci was 0. 730. This suggests that the five microsatellite loci are a valuable tool for further studies about forest musk deer.

Bioleaching of Pb-Zn-Sn chalcopyrite concentrate in tank bioreactor and microbial community succession analysis

The variation of microbial community structure was investigated for the tank bioleaching process of Pb-Zn-Sn chalcopyrite concentrate in the presence of mixed moderately thermophilic bacteria. The parameters, such as pH value, solution potential and concentrations of metal ions, were determined by the method of polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） to analyze the succession of microbial community. The results showed that a final copper extraction rate of 85.6% could be obtained after tank bioleaching for 30 d. The Acidithiobacillus caldus was the dominant population with abundance of about 73.80%in the initial stage, then Sulfobacillus thermosulfidooxidans dominated from the 18th day to the end of bioleaching, while the abundance of Leptospirillum ferriphilum changed slightly. A higher solution potential within a certain range and appropriate concentration of ferric ions were essential for this tank bioleaching of chalcopyrite.

Genetic Relationships Among Four Minorities in Guangxi Revealed by Analysis of 15 STRs

The aim of this study is to investigate the genetic diversity in 15 STRs （short tandem repeats） loci of four minorities in Guangxi Province and to probe into the genetic variation and relationships among these ethnic groups. Allele frequencies of 15 STR loci were collected from 766 unrelated Mulao, Maonan, Miao, and Yao ethnic individuals by PCR-STR and sequencing, and their allele-frequency distribution were compared with each other. The genetic parameters and genetic distances were calculated, and the phylogenetic tree was constructed. Based on the results from this study, 135, 134, 148, and 145 alleles and 424, 432, 445, and 436 genotypes for 15 STR loci were observed in the Mulao, Maonan, Miao, and Yao minorities, respectively. The average heterozygosity of all ethnic groups analyzed was above 0.7; the cumulative power of discrimination （DP）, the probabilities of paternity exclusion （EPP）, and the polymorphic information content （PIC） were greater than 0.99999. Comparison of the allele-frequency distribution indicated that there were significant differences at most loci between Maonan vs. Miao, Yao vs. other groups, but no distinct differences between Mulao vs. Maonan, and Mulao vs. Miao minorities. The NJ tree based on the genetic distance showed that the four minorities were separated into two groups. Mulao and Maonan were clustered into one group, whereas Miao and Yao into the other. Our results revealed that 15 STR loci of the four minorities possessed high genetic diversities. Therefore, the combination of these 15 STRs is a powerful tool for forensic individual identification and paternity investigation, as well as anthropologic and genetic researches. The genetic variation and relationships among the 4 populations revealed by 15 STRs are basically consistent with their linguistic culture and ethical history.

Study on Polymorphisms in the Blood Protein of Tibetan Mastiff

[ Objective] The aim was to study the protein polymorphism in the blood of Tibetan Mastiff, and provide some theoretical basis for resource protection and reasonable development and utilization of Tibetan Mastiff varieties. [ Method] A total of 103 blood samples were taken from four populations of Hequ Tibetan Mastiff, Qinhai Tibetan Mastiff, Tibetan Spaniel and native dogs of Qinghai. Seven blood protein Iocus（Tf, Po, Sα2, Hb, AIb, Pr and Amy）were investigated by using vertical polyacrylamide gel electrophoresis with discontinuous buffer system. Then the genetic variation during different populations was analyzed. [ Result] Genetic variations were observed in Tf, Sα2 and Po in four populations, others were not polymorphic. There were three alleles at the locus of Tf and Po, two alleles at the loci of Sα2. Effective number of alleles and Nei＇s average expected heterozygosity were 1. 532 4 and 0.230 3 relatively, all higher in Tibetan Mastiff than other populations. [ Conclusion] Protein locus in blood of Tibetan Mastiff existed in genetic variation.

Adiponectin Gene Variation -4522C/T Is Associated with Type 2 Diabetic Obesity and Insulin Resistance in Chinese

The authors investigated the possible association of -4522C/T variation of adiponectin gene with coronary heart disease （CHD） and type 2 diabetes mellitus （T2DM）. Genotyping of SNP --4522C/T in 304 patients with CHD, 389 patients with T2DM, and 405 age and sex-matched healthy control subjects was carried out by means of PCR-RFLP approach. No significant difference in the genotype or allele frequencies was found, either between patients with CHD and control subjects, or between patients with T2DM and control subjects. However, in the subgroup analysis, an association of the TAr genotype and T allele with type 2 diabetes combined with obesity （BMI ≥ 25 kg/m2） was found （P = 0.014 and P = 0.034, respectively）. Also the homeostasis model assessment of insulin resistance （HOMA-IR） in T2DM patients with T/T genotype was significantly higher than that in T2DM patients carrying C allele （P = 0.0069）. The authors＇ findings for the first time demonstrated that SNP --4522 in the adiponectin gene was associated with T2DM that combined with obesity and higher insulin resistance index in patients with T2DM. This indicated that the variation might associate with an increased susceptibility to type 2 diabetic obesity and insulin resistance. But -4522C/T polymorphism did not contribute to the susceptibility of CHD.

Characterization of Genetic Polymorphism of Novel MHC B-LBⅡ Alleles in Chinese Indigenous Chickens

Genetic polymorphism of the major histocompatibility complex （MHC） B-LBⅡ gene was studied by amplification of exon 2 using PCR, followed by cloning and DNA sequencing in eight indigenous Chinese chicken populations. To reveal the genetic variation of the B-LB Ⅱ gene, 37 types of patterns detected by PCR-SSCP were investigated first, which would be used to screen novel B-LB Ⅱsequences within the breeds. The types of PCR-SSCP patterns and final sequencing allowed for the identification of 31 novel MHC B-LBⅡ alleles from 30 unrelated individuals of Chinese chickens that were sampled. These are the first designators for the alleles of chicken MHC B-LBⅡ gene based on the rule of assignment for novel mammalian alleles. Sequence alignment of the 31 B-LB Ⅱ alleles revealed a total of 68 variable sites in the fragment of exon 2, of which 51 parsimony informative and 17 singleton variable sites were observed. Among the polymorphic sites, the nucleotide substitutions in the first and second positions of the codons accounted for 36.76% and 35.29%, respectively. The sequence similarities between the alleles were estimated to be 90.6%-99.5%. The relative frequencies of synonymous and nonsynonymous nucleotide substitutions within the region were 2.92%±0.94% and 14.64%±2.67%, respectively. These results indicated that the genetic variation within exon 2 appeared to have largely arisen by gene recombination and balancing selection. Alignment of the deduced amino acid sequences of the β1 domain coded by exon 2 revealed 6 synonymous mutations and 27 nonsynonymous substitutions at the 33 disparate sites. In particular, the nonsynonymous substitutions at the putative peptide-binding sites are considered to be associated with immunological specificity of MHC B-LB Ⅱ molecule in Chinese native chickens. These results can provide a molecular biological basis for the study of disease resistance in chicken breeding.

Polymorphism and Morphotype Transformations in the Rotifer(Brachionus calyciflorus)

The relationship of three Brachionus calyciflorus morphotypes （two short-spined, single-spined and spineless ones） was studied in the laboratory. Newly hatched females of each morphotype were cultured individually in an inorganic or Asplanchna-conditioned mediums, and the morphotype of their offspring were checked at 200 x magnification. Females of each morphotype, cultured in the inorganic medium, could produce offspring of these three morphotypes in a single generation. Females of all three morphotyes could respond to specific Asplanchna-substances when they were cultured in Asplanchna-conditioned medium. Moreover, we observed that these three morphotype females could coexist in the medium and had three types of egg-bearing females. Based on the above observations we suggest that B. calyciflorus was a morphotype complex and has three basic morphotypes.