AGTR1 A1166C gene polymorphism is associated with the effectiveness of valsartan monotherapy in Chinese patients with essential hypertension:A retrospective analysis

Objective:To investigate whether angiotensinⅡtype 1 receptor(AGTR1 A1166C)gene polymorphism was associated with the effectiveness of valsartan monotherapy in Chinese patients with essential hypertension.Methods:This retrospective analysis included 198 patients(≥18 years of age)who received valsartan monotherapy(80 mg/day)for newly developed essential hypertension at the authors’center between January 1,2020 and December 31,2023.Genotyping for AGTR1 A1166C gene polymorphism was done by polymerase chain reaction(PCR)-melting curve analysis of genomic DNA from peripheral blood samples.A dominant genetic model for AGTR1 A1166C(AA genotype versus AC+CC genotype)was used.Multivariate regression analysis of baseline variables and AGTR1 polymorphism was conducted to identify predictors of target blood pressure attainment(<140/90 mmHg)at the 4-week follow-up.Results:The median age of the 198 patients was(53.7±13.5)years,and 58%were men.Genotyping assays showed that 164 patients had the AA genotype,and 34 patients were of the AC/CC genotype,including 30 with the AC genotype and 4 with the CC genotype.Allele distribution was consistent with Hardy Weinberg equilibrium.109 Patients(55.1%)attained the blood pressure target.Multivariate analysis showed that smoking(versus no smoking,HR 0.314,95%CI 0.159-0.619,P=0.001)and AGTR1 A1166C AA genotype(versus AC/CC,HR 2.927,95%CI 1.296-6.611,P=0.023)were significant and independent predictors of target attainment.25 Patients(73.5%)with AGTR1 A1166C AC/CC genotype attained the target versus 51.2%(51/164)of patients with AGTR1 A1166C AA genotype(P=0.017).Patients with AGTR1 A1166C AC/CC genotype had a significantly greater reduction in systolic blood pressure[(33.1±10.8)mmHg versus(29.2±11.7)mmHg in AA carriers;(P=0.029)].Conclusions:Hypertensive patients carrying one or two C alleles of the AGTR1 A1166C gene were more responsive to valsartan treatment.

双能CT联合血清PIVKA-Ⅱ、NDRG4诊断卵巢癌效能

目的:探究双能CT联合血清拮抗剂-Ⅱ诱导的蛋白质(PIVKA-Ⅱ)、抑癌基因N-myc下游调节因子4(NDRG4)检测在诊断卵巢癌中的应用价值。方法:2020年2月-2023年5月本院接受治疗的卵巢癌患者105例为卵巢癌组,同期收治的良性卵巢肿瘤患者100例为良性组,健康体检者90例为对照组。所有受试者均行双能CT检查,测量双能CT参数标准化碘浓度(NIC)和能谱曲线斜率(k)值,检测血清PIVKA-Ⅱ、NDRG4水平。采用受试者工作特征(ROC)曲线分析双能CT参数联合血清PIVKA-Ⅱ、NDRG4的诊断卵巢癌价值;Pearson法分析血清PIVKA-Ⅱ、NDRG4与双能CT参数的相关性。结果:对照组、良性组、卵巢癌组NIC、k值、血清PIVKA-Ⅱ水平依次升高,NDRG4依次降低;双能CT参数NIC、k及血清PIVKA-Ⅱ、NDRG4诊断卵巢癌的曲线下面积(AUC)分别为0.785、0.696、0.832、0.799,4项联合诊断卵巢癌的AUC(0.937)显著提高(均P<0.05)。卵巢癌患者血清PIVKA-Ⅱ水平与NIC、k呈正相关,血清NDRG4水平与NIC、k呈负相关;双能CT参数NIC、k、血清PIVKA-Ⅱ、NDRG4水平与患者FIGO分期、淋巴结转移、分化程度有关(均P<0.05)。结论:双能CT、血清PIVKA-Ⅱ、NDRG4对卵巢癌诊断具有一定价值,且联合诊断价值更高。

胃窦癌组织中LAG-3 FGL1 MHC-Ⅱ的表达与预后的关系

目的:探索新型免疫检查点淋巴细胞激活基因3(lymphocyte-activation gene 3,LAG-3)、纤维蛋白原样蛋白1(fibrinogenlike protein 1,FGL1)、主要组织相容性复合体Ⅱ类分子(major histocompatibility complex classⅡ,MHC-Ⅱ)在胃窦癌(gastric antral cancer,GAC)中的表达情况与预后的相关性。方法:收集2012年1月至2014年12月于天津医科大学肿瘤医院诊断为GAC的67例患者病理标本,分别进行石蜡切片制作,采用免疫组织化学法检测LAG-3、FGL1、MHC-Ⅱ三个指标的表达情况,并用统计学方法分析组间差异。采用Kaplan-Meier法评估LAG-3、FGL1、MHC-Ⅱ的表达水平与GAC患者预后之间的关系并绘制生存曲线。结果:GAC患者中,肿瘤大小<4 cm的患者和无淋巴结转移的患者LAG-3免疫细胞阳性率更高(P<0.05);女性患者MHC-Ⅱ免疫细胞阳性率更高(P<0.05)。免疫细胞中LAG-3、MHC-Ⅱ高表达的患者总生存期(overall survival,OS)较好(P<0.05);肿瘤细胞中MHC-Ⅱ高表达的患者OS、无病生存期(disease-free survival,DFS)较差(P<0.05);而FGL1在免疫细胞和肿瘤细胞中的表达与OS、DFS无显著相关性(P>0.05)。结论:GAC患者LAG-3、MHC-Ⅱ在不同区域的表达量存在差异,GAC患者LAG-3及其配体在免疫细胞的表达对预后产生积极影响,提示免疫细胞中LAG-3/MHC-Ⅱ可以作为GAC患者预后标志物,为临床个体化免疫治疗提供新的依据。

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.

山羊CAST基因Ⅱ型转录本的克隆及在山羊不同组织中的表达

钙蛋白酶抑制蛋白（Calpastatin，CAST）基因是与畜禽肉质性状密切相关的重要候选基因。文章根据牛和绵羊CAST基因mRNA，应用RACE技术首次成功克隆了山羊CAST基因Ⅱ型转录本（以下简称CASTII基因）全长cDNA，对序列及编码的氨基酸进行了生物信息学分析。结果显示，该基因cDNA全长2474bp，完整的开放阅读框（ORF）为558～2252bp，编码564个氨基酸。氨基酸序列中存在4个保守结构域和1个保守七肽序列；蛋白质二级结构以无规卷曲和a-螺旋为主，富含疏水区，存在多个磷酸化位点以及蛋白激酶C（Protein kinaseC，PKC）的磷酸化位点。通过实时荧光定量RT—PCR技术分析了CASTⅡ基因在天府肉羊部分组织中的表达情况。结果表明：CASTII基因在所选择的天府肉羊7种组织中均有表达，半岁各组织中，眼肌的表达量最高，与腿肌差异显著（P〈0．05），极显著高于内脏各组织（P〈0．01）；在眼肌组织中，CASTⅡ基因的表达量随着年龄的增长而增加，3岁时的表达量最高。

细胞色素氧化酶P4502C19基因多态检测联合血清肝素辅助因子Ⅱ对下肢动脉硬化闭塞症介入术后再狭窄的预测价值

目的探讨细胞色素氧化酶P4502C19(CYP2C19)基因多态检测联合血清肝素辅助因子Ⅱ(HCⅡ)对下肢动脉硬化闭塞症(ASO)介入术后再狭窄的预测价值。方法收集2017年1月至2021年5月于绵阳市第三人民医院行介入治疗的146例ASO患者的临床资料,根据随访期间再狭窄发生情况将其分为再狭窄组(n=27)和无再狭窄组(n=119)。收集患者性别、年龄、体重指数(BMI)、吸烟史、心脑血管病史、高同型半胱氨酸病史、病变血管支数、术前狭窄情况、支架植入情况、纤维蛋白原水平、总胆固醇水平、红细胞计数、CYP2C19基因多态性、HCⅡ活性,分析ASO介入术后再狭窄的影响因素。结果有吸烟史、支架植入、CYP2C19慢代谢型及HCⅡ活性均为ASO支架植入术后发生再狭窄的独立危险因素(P﹤0.05)。受试者工作特征(ROC)曲线结果显示,HCⅡ活性预测ASO支架植入术后再狭窄的截断值为95.80%,曲线下面积(AUC)为0.809(95%CI:0.719~0.900),灵敏度为74.80%,特异度为88.24%,Kappa=0.566。CYP2C19基因多态检测对ASO支架植入术后再狭窄的预测灵敏度为66.67%,特异度为89.07%,Kappa=0.537。联合检测对ASO支架植入术后再狭窄的预测灵敏度为96.30%,特异度为86.55%,Kappa=0.682。结论ASO介入术后再狭窄与吸烟史、支架植入情况、CYP2C19基因多态性及HCⅡ活性有关,CYP2C19基因多态性及HCⅡ活性检测均可用于预测ASO介入术后再狭窄,但联合检测可提高其诊断效能。

Expression of IGF-Ⅱ,p53,p21 and HBxAg in precancerous events of hepatocarcinogenesis induced by AFBI and/or HBV in tree shrews

INTRODUCTIONIn order to study the relationship between oncogeneexpression and HCC generation,we observed theprecancerous hepatic GGT loci,IGF-Ⅱ,p53 andp21 expression during hepatocarcinogenesis of treeshrew induced by hepatitis B virus (HBV) and/oraflatoxin B1 (AFB1).

Association of Polymorphisms in Angiotensin-converting Enzyme and Type 1 Angiotensin Ⅱ Receptor Genes with Coronary Heart Disease and the Severity of Coronary Artery Stenosis

To explore the relation of angiotensin-converting enzyme （ACE） and angiotensin Ⅱ type 1 receptor （AT1R） gene polymorphism with coronary heart disease （CHD） and the severity of coronary artery stenosis, 130 CHD patients who underwent coronary angiography were examined for the number of affected coronary vessels （≥75% stenosis） and coronary Jeopardy score. The insertion/deletion of ACE gene polymorphism and AT1R gene polymorphism （an A→C transversion at nucleotide position 1166） were detected by using polymerase chain reaction （PCR） and restriction fragment length polymorphism （RFLP） in CHD patients and 90 healthy serving as controls. The resuits showed that DD genotype and of ACE were more frequent in CHD patients than that in control group （38.5% vs 14.4%, P〈0.001）. The frequency of the ATIR A/C genotypes did not differ between the patients and the controls （10% vs 13.1%, P〉0.05）. The relative risk associated with the ACE-DD was increased by AT1R-AC genotype. Neither the number of affected coronary vessels nor the coronary score differed among the ACE I/D genotypes （P〉0.05）. But the number of affected coronary vessels and the coronary score were significantly greater in the patients with the AT1R-AC genotype than in those with the AA genotype （P〈0.05）. In conclusion, DD genotype may be risk factor for CHD and MI in Chinese people, and is not responsible for the development of the coronary artery stenosis. The AT1R-C allele may increase the relative risk associated with the ACE-DD genotype, and may be involved in the development of the stenosis of coronary artery.

Genetic Differentiation Analyses Based on mtDNA COⅡ Gene Sequences Among Different Geographic Populations of Aphis glycines(Hemiptera: Aphididae) in Northeast China

Aphis glycines （Hemiptera： Aphididae） is considered as a cosmopolitan pest of cultivated soybean, major difficulties in its control measures may be due to its higher genetic diversity; however, the knowledge about population genetic diversity of this species is limited. This study aimed to represent the genetic differentiation among different geographic populations of soybean aphid in Northeast China. In order to investigate and assess the genetic diversity, genetic differentiation, molecular variance, population structure, ecological importance and evolutionary history of A. glycines, we sequenced a fragment of one protein-coding gene, the cytochrome c oxidase I/of mitochondrial DNA gene. The results showed that four haplotypes were defined among CO 11 gene of 180 sequences of soybean aphid in Northeast China including H1 shared by all the populations. Lower haplotype diversity （Hd=0.3590± 0.0420） and nucleotide diversity （Pi=0.0012±0.0002） were observed and high gene flow was detected in every two populations, while most of the variation （80.81%） arose from variability within A. glycines from individuals. Low genetic differentiation and high gene flow （Nm=2.106） indicated a high migration rate between the populations, which might reveal that gene flow in different geographic populations did not affect by geographical distance. The phylogenetic tree and the haplotype network ofA. glycines were obtained based on sequences of CO Ⅱ gene, there were no significant genealogical branches or clusters recognized in NJ tree, and no clear distribution, delineation of haplotypes were demonstrated in the haplotype network according to geographical location. This study rejected the vicariance hypothesis： geographic isolation could be a barrier and it restricted A. glycines gene flow among 10 populations.

Genetics of coronary heart disease with reference to ApoAICⅡI-AIV gene region

Cardiovascular diseases are affected by multiple factors like genetic as well as environmental hence they reveal factorial nature. The evidences that genetic factors are susceptible for developing cardiovascular diseases come from twin studies and familial aggregation. Different ethnic populations reveal differences in the prevalence coronary artery disease(CAD) pointing towards the genetic susceptibility. With progression in molecular techniques different developments have been made to comprehend the disease physiology. Molecular markers have also assisted to recognize genes that may provide evidences to evaluate the role of genetic factors in causation of susceptibility towards CAD. Numerous studies suggest the contribution of specific "candidate genes", which correlate with various roles/pathways that are involved in the coronary heart disease. Different studies have revealed that there are large numbers of genes which are involved towards the predisposition of CAD. However, these reports are not consistent. One of the reasons could be weak contribution of genetic susceptibility of these genes. Genome wide associations show different chromosomal locations which dock, earlier unknown, genes which may attribute to CAD. In the present review different ApoAI-CⅡI-AIV gene clusters have been discussed.

Relationship between polymorphism of class Ⅱ transactivator gene promoters and chronic hepatitis B

AIM: To investigate the relationship between the polymorphism of class Ⅱ transactivator (CIITA) gene promoters and chronic hepatitis B (CHB). METHODS: Genomic DNA was prepared from peripheral blood leukocytes. Promoters Ⅰ,Ⅲ and Ⅳ of gene were analyzed respectively with polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) in 65 patients with CHB, 26 patients with acute hepatitis B (AHB) and 85 normal controls. RESULTS: No abnormal migration was found in PCR-SSCP analysis of the three promoters in the three groups. Also, no sequential difference was observed at the three promoters among the CHB patients, AHB patients and normal controls. CONCLUSION: No polymorphism in promoters I, III and IV of CIITA gene exists in CHB patients, ABH patients and normal controls, suggesting that the promoter of CIITA gene might be a conserved domain.

Cloning of Human Uroplakin Ⅱ Gene from Chinese Transitional Cell Carcinoma of Bladder and Construction of Its Eukaryotic Expression Vector

Summary: To clone Uroplakin Ⅱ gene from Chinese transitional cell carcinoma (TCC) of bladder and construct its eukaryotic expression vector, the molecular cloning method was used to extract total RNA from a GⅢ/ T 3N 0M 0 tissue sample of the bladder TCC patients. The primers were designed by Primer 5.0 software. Full length cDNA of Uroplakin Ⅱ gene was amplified by reverse transcription polymerase chain reaction (RT-PCR), assayed by nucleic acid sequencing and then inserted between XbaⅠ and HindⅢ restrictive sites of eukaryotic expression vector pcDNA3.0. The recombinant was assayed by restricted enzyme digestion. Under the induction of Lipofectamine 2000, the recombinant was transfected into Uroplakin Ⅱ negative bladder cancer cell line EJ. Cellular expression levels of Uroplakin Ⅱ were detected by RT-PCR. The nucleic acid sequencing results indicated that Chinese Uroplakin Ⅱ cDNA (555 bp) was successfully cloned. The BLAST analysis demonstrated that the cloned sequence is 100 % homologous with sequences reported overseas. The GenBank accession number AY455312 was also registered. The results of restricted enzyme digestion indicated that eukaryotic vector pcDNA-UPⅡ for Uroplakin Ⅱ was successfully constructed. After being transferred with pcDNA-UPⅡ for 72 h, cellular Uroplakin Ⅱ mRNA levels were significantly improved (P<0.01). It is concluded that human Uroplakin Ⅱ gene was successfully cloned from Chinese TCC tissues, which provided a basis for further exploration of the roles of Uroplakin Ⅱ gene in TCC biological behaviors and potential strategies for targeted biological therapy of TCC.

Structural analysis of DMD gene and its clinical application in Chinese.Ⅰ. Bgl Ⅱ exon-containing fragment,RFLP and carrier detection

This article is one of the serial studies oll the characteristics of the molecular structure for dystrophin gene in Chinese. By using the entire dystrophin cDNA (14 kb) as a probe- the number and RFLPs of Bgl Ⅱ exon-containing fragments of the dystrophin gene were analysed. Four new Bgl Ⅱ fragments were found, two of them (3.7 and 6.2 kb) detected by comparing the hybridization patterns with cDNA1-2a. 1a and 2a, one (9.3 kb) from the hybridization pattern with cDNA 9 by lengthening migrating distance of DNA fragments in electrophoresis. and another one (4.0 kb) by comparing the patterns with cDNA 11-14,11a- 11b’ 11c-12a and 14. The results indicated that the number of Bgl Ⅱ exon-containing fragments should be 59 rather than 55 reported previously, which laid the foundation of the Bgl Ⅱ partial restriction map for dystrophin gene. Three of the four RFLPs found in Caucacian appear in the hybridization patterns of three subclones, i.e.cDNA 2b-3. cDNA 4-5, and cDNA 5b-7.’ The values of expected heterozygote frequency (EHF) were 0.33, 0.33and 0.40 and the observed heterozygote frequency (OHF)were 0.40. 0.40 and 0.48 respectively. Meanwhile, two new rare allelic fragments (15 kb) were found in RFLPs from Bgl Ⅱ/2b-3 and Bgl Ⅱ/4-5a patterns respectively. These Bgl Ⅱ RFLPs and four XbaI RFLPs documented in our laboratory, have been used to detect the carrier in 7 DMDfamilies and 1 BMD family. Of the 69 individuals from the 8 families- 11 females were diagnosed as the carriers with DMD mutation, 4 females as the doubtful carriers, 12 females were defined as normal genotype and 2 females as probably normal. The results suggest that the carrier testing method based on dosage intensity analysis and genotype analysis by using dystrophin cDNA as a probe will be more sensitive and accurate.

Allelic Polymorphism,Gene Duplication and Balancing Selection of MHC Class ⅡB Genes in the Omei Treefrog(Rhacophorus omeimontis)

The worldwide declines in amphibian populations have largely been caused by infectious fungi and bacteria. Given that vertebrate immunity against these extracellular pathogens is primarily functioned by the major histocompatibility complex（MHC） class Ⅱ molecules, the characterization and the evolution of amphibian MHC class Ⅱ genes have attracted increasing attention. The polymorphism of MHC class Ⅱ genes was found to be correlated with susceptibility to fungal pathogens in many amphibian species, suggesting the importance of studies on MHC class Ⅱ genes for amphibians. However, such studies on MHC class Ⅱ gene evolution have rarely been conducted on amphibians in China. In this study, we chose Omei treefrog（Rhacophorus omeimontis）, which lived moist environments easy for breeding bacteria, to study the polymorphism of its MHC class Ⅱ genes and the underlying evolutionary mechanisms. We amplified the entire MHC class ⅡB exon 2 sequence in the R. omeimontis using newly designed primers. We detected 102 putative alleles in 146 individuals. The number of alleles per individual ranged from one to seven, indicating that there are at least four loci containing MHC class ⅡB genes in R. omeimontis. The allelic polymorphism estimated from the 102 alleles in R. omeimontis was not high compared to that estimated in other anuran species. No significant gene recombination was detected in the 102 MHC class ⅡB exon 2 sequences. In contrast, both gene duplication and balancing selection greatly contributed to the variability in MHC class ⅡB exon 2 sequences of R. omeimontis. This study lays the groundwork for the future researches to comprehensively analyze the evolution of amphibian MHC genes and to assess the role of MHC gene polymorphisms in resistance against extracellular pathogens for amphibians in China.

Autosomal dominant osteopetrosis typeⅡresulting from a de novo mutation in the CLCN7 gene:A case report

BACKGROUND Osteopetrosis is a family of extremely rare diseases caused by failure of osteoclasts and impaired bone resorption. Among them, autosomal dominant osteopetrosis type Ⅱ(ADO Ⅱ), related to the chloride channel 7(CLCN7) gene, is the most frequent form of osteopetrosis. In this study, we report a de novo mutation of CLCN7 in a patient without the family history of ADO Ⅱ.CASE SUMMARY A 5-year-old Chinese boy with ADO Ⅱ was found to have a de novo mutation in the CLCN7 gene [c.746 C>T(p.P249 L)]. Typical clinical manifestations, including thickening of the cortex of spinal bones and long bones, non-traumatic fracture of the femoral neck, and femoral head necrosis, were found in this patient. The patient is the first reported case of ADO Ⅱ with the missense mutation c.746 C>T(p.P249 L) of the CLCN7 gene reported in China. We also review the available literature on ADO Ⅱ-related CLCN7 mutations, including baseline patient clinical features, special clinical significance, and common mutations.CONCLUSION Our report will enrich the understanding of mutations in ADO Ⅱ patients. The possibility of a de novo mutation should be considered in individuals who have no family history of osteopetrosis.

Mutations in Exons of the CYP17-Ⅱ Gene Affect Sex Steroid Concentration in Male Japanese Flounder(Paralichthys olivaceus)

As a specific gene of fish,cytochrome P450c17-Ⅱ(CYP17-Ⅱ) gene plays a key role in the growth,development and reproduction level of fish.In this study,the single-stranded conformational polymorphism(SSCP) technique was used to characterize polymorphisms within the coding region of CYP17-Ⅱ gene in a population of 75 male Japanese flounder(Paralichthys olivaceus).Three single nucleotide polymorphisms(SNPs) were identified in CYP17-Ⅱ gene of Japanese flounder.They were c.G594A(p.G188R),c.G939A and c.G1502A(p.G490D).SNP1(c.G594A),located in exon 4 of CYP17-Ⅱ gene,was significantly associated with gonadosomatic index(GSI).Individuals with genotype GG of SNP1 had significantly lower GSI(P < 0.05) than those with geno-type AA or AG.SNP2(c.G939A) located at the CpG island of CYP17-Ⅱ gene.The mutation changed the methylation of exon 6.Indi-viduals with genotype AA of SNP2 had significantly lower serum testosterone(T) level and hepatosomatic index(HSI) compared to those with genotype GG.The results suggested that SNP2 could influence the reproductive endocrine of male Japanese flounder.How-ever,the SNP3(c.G1502A) located in exon 9 did not affect the four measured reproductive traits.This study showed that CYP17-Ⅱ gene could be a potentially useful candidate gene for the research of genetic breeding and physiological aspects of Japanese flounder.

1例罕见α-地中海贫血产前诊断与家系分子遗传学分析

目的 对1例疑似携带罕见地中海贫血(简称地贫)的产前诊断胎儿进一步测序分析,对先证者进行家系分子遗传学分析。方法 运用血常规和血红蛋白电泳进行地贫筛查,采用gap-PCR法和PCR-RDB法检测24种地贫突变,对疑似罕见地贫进行基因测序分析。结果 先证者为--~(SEA)地贫与α2基因IVS-Ⅱ-119地贫双重杂合子,其IVS-Ⅱ-119地贫基因遗传自母方,--~(SEA)地贫基因遗传自父方。结论 --~(SEA)/α~(IVS-Ⅱ-119)α HbH地贫患儿的诊断,为罕见地贫的遗传咨询和产前诊断提供科学理论依据。

丹参素和川芎嗪对血管紧张素Ⅱ致心肌肥大相关基因的影响

目的通过研究活血药的提取物丹参素和川芎嗪对血管紧张素Ⅱ(AngⅡ)致心肌肥大及相关基因的影响,探讨其抑制心肌肥大的作用机理。方法以心钠素(ANP)和肌动蛋白- β(β- actin)基因表达为指标,采用一步法,应用TRIzolReagent提取心肌细胞总RNA ,然后用RT PCR方法测定其ANP、β- actin的mRNA表达。结果分子生物学研究表明,AngⅡ可显著增加心肌细胞ANPmRNA的表达(P <0 . 0 1) ,而Losartan可明显抑制AngⅡ所诱导的ANPmRNA的表达(P <0 . 0 1) ,丹参素和川芎嗪也可减少ANPmRNA的表达(P <0 . 0 5 ) ;AngⅡ同时也增加心肌细胞-βactinmRNA的表达,而Losartan、丹参素和川芎嗪可显著抑制其表达(P <0 .0 5 )。结论活血药的有效组分丹参素和川芎嗪可抑制AngⅡ对心肌细胞ANP和β-actin基因表达的增加,具有防止心肌细胞肥大作用,从而防治心脏肥厚。

大肠杆菌L－天门冬酰胺酶Ⅱ基因的高效表达

用一对与大肠杆菌天门冬酰胺酶Ⅱ基因（ＡｎｓＢ）两侧序列互补的寡核苷酸Ｅ１，Ｅ２作引物，以大肠杆菌野生株ＣＰＵ２１０００９的染色体ＤＮＡ为模板，通过ＰＣＲ扩增得到约１．２ｋｂ的ＤＮＡ片段，将此片段插入ｐＫＫ２３３－２的ｔａｃ启动子下游，转化不同的大肠杆菌宿主菌株，结果表明以ＣＰＵ２１０００９为宿主菌时，转化子的酶表达量最高，重组Ｌ－天门冬酰胺酶占菌体总蛋白４８．３％，发酵液酶活力达４００ＩＵ／ｍｌ；比活为４８ＩＵ／ｍｇ蛋白，ＳＤＳ－ＰＡＧＥ显示纯化后的重组Ｌ－天门冬酰胺酶分子量与天然酶相同，均为１４０ＫＤ。

丹参酮ⅡA对猪主动脉内皮细胞受血管紧张素Ⅱ作用时产生NO及eNOS基因表达的影响

目的探讨丹参酮ⅡA对血管内皮细胞的保护作用。方法采用硝酸还原酶法、逆转录聚合酶链式反应(RT-PCR)和免疫组织化学法,分别检测不同作用时间(1h、6h、24h)的血管紧张素Ⅱ(AngⅡ)以及在AngⅡ作用的不同时间点(0h点为A组、6h点为B组)加入丹参酮ⅡA对培养的猪主动脉内皮细胞产生NO及其内皮型一氧化氮合酶(eNOS)的蛋白和mRNA表达。结果(1)随着AngⅡ作用时间的延长,血管内皮细胞NO的产生及eNOS的表达呈显著下降(P<0.01),表现出时间依赖性的负性作用。(2)丹参酮ⅡA可抑制AngⅡ对内皮细胞分泌NO及eNOS基因表达的负性作用(P<0.01)。(3)在丹参酮ⅡA作用1h、6h,A组的抑制效应明显强于B组(P<0.05);随着作用时间延长至24h,两组比较差异无显著性。结论丹参酮ⅡA可抑制AngⅡ对血管内皮细胞分泌NO以及细胞eNOS基因表达的负性作用。