P1PK血型不合引起胎死宫内的罕见病例研究1例

目的分析1例胎死宫内患者血型血清学及分子生物学特点,明确患者血型及不规则抗体类型。方法采用血清学方法对患者的ABO、P1PK血型及不规则抗体进行鉴定;对编码P1PK抗原形成关键酶的α1,4-半乳糖基转移酶基因(α1,4-galactosyltransferase gene,A4GALT)进行测序,分析DNA序列,确定突变位点。结果血清学实验证实患者为P1PK血型系统的p表型,患者血清中存在抗-PP1PK;测序结果显示,患者的A4GALT编码区出现304-305insG突变。结论患者为P1PK血型系统的p表型,其A4GALT基因突变类型为304-305insG,患者体内的抗-PP1PK可能为其胎死宫内的原因。     

p血型引起交叉配血不合一例分析

细胞的抗体,该试验是输血前相容性试验中防止发生错误的最后一道“安全线”,严格的交叉配血需能够检出ABO不相容及 ABO系统之外的有临床意义的抗体,主要为 IgG类抗体。     

一例p表型的血清学分析

目的分析患者血型血清学特点,明确患者血型及血型抗体特异性,为患者寻找配合型血液。方法采用血型血清学方法进行红细胞ABO、p血型鉴定;采用盐水法、微柱凝胶法、Liss-IAT法进行不规则抗体筛查和抗体特异性鉴定。结果患者血型为B型,p表型,血清中检出抗-PP1PK。结论患者为罕见p表型,血清中含有抗-PP1PK。     

血清学和分子生物学鉴定Lewis血型抗体引起的输血反应

为了分析1例溶血性输血反应产生的原因，用谱细胞和Le（a—b-）表型细胞鉴定患者血清中的抗体，用抗Le^a和抗Le^b血型试剂鉴定患者红细胞表型，用PCR测序方法分析LE基因（FUT3基因）全编码区序列。结果表明：患者血清中有抗Le^a和抗Lc^b抗体，血型表型为Le（a—b-），LE基因型为无效等位基因纯合子（1e59，508）。结论：抗Le^b抗体可引起溶血性输血反应，患者FUT3基因突变导致Le（a—b-）表型。     

血清学鉴定罕见Rhnull血型1例

笔者在无偿献血者中发现1例缺失D、C、c、E、e抗原的Rhnull型。国际上有14个家族的22例报道[1]。现将结果报告如下。1材料与方法1.1样本来源本中心无偿献血者,女,32岁,否认输血史及生育史,有2次药物流产史。1.2试剂和仪器单克隆抗-A、-B血清(长春博德公司,批号20050306);IgM抗-D血清(苏州大学医学生物技术研究所,批号20041204);IgG IgM抗-D血清(台湾Baso、德国Biotest、法国Diagast公司),人源IgG抗-D血清(中国医学科学院输血研究所),抗-C、-c、-E、-e血清(德国Biotest公司),DianaGel微柱凝胶卡、WADiana全自动配血系统(Grifols…     

中国彝族人群p表型家系分析及新A4GALT等位基因鉴定

目的 研究一个彝族家系P1Pk血型系统中罕见p表型的分子遗传机制.方法 对34例家系成员样本进行血型血清学鉴定;红细胞稀有血型基因分型采用荧光定量PCR法;同时采用Sanger测序法分析α1,4-半乳糖基转移酶(α-1,4-galactosyltransferase,A4GALT)基因和β1,3-N乙酰半乳糖胺转移酶(...     

罕见B(A)血型的鉴定

目的观察2例罕见B(A)血型的血清学特征并研究其分子机制。方法用血型血清学鉴定2例献血者标本ABO血型,用序列特异性引物聚合酶链反应(PCR-SSP)基因分型和直接测序确定其基因型。结果 2例献血者血型血清学检测结果均表现为B(A)亚型的特点。标本1基因分型为BO2,测序结果为第7外显子B基因发生640AG突变,符合B(A)04/O02的基因型特点;标本2基因分型为BO1,测序结果为第7外显子B基因发生700CG突变,符合B(A)02/O01的基因型特点。结论2例标本均为B(A)表现型,基因型分别为B(A)04/O02和B(A)02/O01。     

罕见B(A)血型的血清学及基因检测研究

本研究旨在通过血清学和分子生物学方法研究1例ABO血型系统中罕见B(A)型,并分析造成多次检测结果不一致的原因。采用血型血清学方法鉴定ABO血型血清型,同时采用PCR测序方法检测其基因型。结果表明,该献血者血清学结果正反定型结果不符,正定型AB型,反定型B型,ABO基因型为B(A)04/O01型,存在nt640A>G点突变,导致214位甲硫氨酸被缬氨酸置换。结论:对ABO血型血清学正反定型不符的标本,可用分子生物学方法进行辅助验证,并能阐述ABO血型弱表达现象的分子基础。     

血清学与分子生物学技术鉴定产生抗-Dib抗体的Di(a+b-)血型1例

目的通过血清学实验和分子生物学技术鉴定出1例产生抗-Dib抗体的Di(a+b-)稀有血型。方法采用血清学方法对患者进行血型鉴定、直抗试验和不规则抗体筛查及鉴定试验,采用DNA测序方法鉴定患者的Diego血型基因。结果患者体内存在抗高频抗原抗体,分子水平确定其为Di(a+b-)稀有血型,因此其抗体为抗-Dib抗体。结论分子生物学鉴定血型基因有助于抗高频抗原抗体的鉴定。     

一例稀有血型JK(a-b-)表型家系调查分析

本研究旨在检测Kidd血型系统的稀有JK(a-b-)表型的先证者及其家系中其他个体的表型及基因型,探讨本例JK(a-b-)表型的分子机理,分析遗传背景,为稀有血型个体输血治疗寻找相合供者提供科学依据。采用4 mol/L尿素溶血试验,筛选JK(a-b-)表型先证者,并对其进行家系调查。用血清学方法确认表型;用PCR-SSP方法检测基因型,并对JK基因外显子4-11及其侧翼的区域扩增后测序,分析序列信息。结果表明,先证者及其哥哥具有相同的表型JK(a-b-)和基因型Jkb/Jkb;其父母的表型为JK(a+b-),基因型为Jka/Jkb;妹妹的表型和基因型分别为JK(a+b-)和Jka/Jka。对外显子及其侧翼测序发现,先证者及其哥哥的内含子5的3’端拼接接受位点碱基g>a突变,其父母的该位点均为a/g杂合,其妹妹的该位点没有突变;内含子3(外显子4侧翼)nt-99位的碱基为g,其父母的该位点均为g/a杂合,其妹妹的该位点是a。结论:先证者及其哥哥具有相同的表型JK(a-b-)和基因型Jkb/Jkb,JK基因内含子5的3’端拼接接受位点的g>a突变可能是造成其JK(a-b-)表型的原因之一;家系中内含子3的nt-99位的碱基也具有多态性(ncbi:rs8090908),该位点多态性是否与JK(a-b-)表型相关值得进一步研究。本例家系Kidd血型系统符合显性遗传规律。对于稀有JK(a-b-)表型个体的家系,尤其是同胞兄妹进行血型调查,发现表型和基因型均相同的个体可能性大。     

High lipoprotein(a) levels with predominance of high molecular weight apo(a) isoforms in patients with pulmonary embolism

Abstract. Lipoprotein(a) (Lp(a)) may interact with the cellular components and protein co-factors of fibrino-lysis. To evaluate the effect of Lp(a) in thromboem-bolic diseases of the venous system, we measured serum levels and the isoform distribution of apo(a) in 25 patients with pulmonary embolism (18 men, 7 women, aged 21—77 years). The control group was adjusted for sex and age ( P = 0.189). Serum Lp(a) concentration was significantly higher in the study group (median: 9.3 vs. 4.3 mg dL^-1). As the distribution of high and low molecular weight subtypes of apo(a) did not show any differences ( P = 0.127) between the two groups, the elevated Lp(a) levels in patients with pulmonary embolism could not be attributed to the investigated kringle-4 polymorphism of the apo(a) gene and therefore other genetic or non-genetic implications are indicated.     

Identification of two novel FUT1 mutations in people with Bombay phenotype from Iran

Background and purposeBombay and Para-Bombay phenotypes are characterized by FUT1 gene mutation and lack of H antigen expression in red blood cells. ABH antigens are not present in the body secretions of Bombay individuals, while they are expressed in the secretions of para-Bombay. The aim of this study was to investigate the molecular basis of FUT1 and FUT2 genes in Iranians with the Bombay or Para-Bombay phenotype.Materials and MethodsABO phenotype analysis and routine serological tests were performed on 11 people with Bombay and Para-Bombay phenotypes. The coding regions of FUT1 and FUT2 genes were amplified by PCR followed by sequencing. The ABO genotypes were also determined by sequencing exons 6 and 7 of the ABO gene. Results: Serological investigations confirmed the Bombay phenotype in 8 samples and the Para-Bombay phenotype in 3 samples. Family members with the Bombay phenotype had the classic c 0.725 T > G mutation in the FUT1 gene, accompanied by deletion of the FUT2 gene. Other samples had c.653 A>G, c 0.661 C>T, c 0.652 C>G, and c.722 A>C mutations in the FUT1 while FUT2 was silenced by c 0.461 G>A. Conclusion: In this research, we identified two novel mutations in the FUT1 gene in individuals with the Bombay phenotype. This and previous works confirm the variety of FUT1 mutations.     

Relationship between lipoprotein(a) phenotypes and albumin excretion rate in non-insulin-dependent diabetes mellitus: protective effect of ‘null’ phenotype?

The possible association between lipoprotein(a) [Lp(a)] and albumin excretion rate (AER) is a topic that generates conflicting views. In addition, Lp(a) phenotypes have not previously been considered as factors influencing AER. In order to clarify this issue, we studied 70 non-insulin-dependent diabetes mellitus (NIDDM) patients without clinically detectable macroangiopathy, 27 with microalbuminuria and 43 without it. Both groups were matched for the known variables that could influence AER and serum Lp(a) levels. Lp(a) was determined by enzyme-linked immunosorbent assay (ELISA), and Lp(a) phenotypes were assessed by electrophoresis followed by immunoblotting. Lp(a) phenotypes were grouped as follows: 'small' (F, S1 and S2), 'big' (S3 and S4) and 'null'. The NIDDM patients with microalbuminuria presented higher serum Lp(a) concentrations than the patients without it [15.7 mg dL⁻¹ (95% CI 0.5–36.5) vs. 4.5 mg dL⁻¹ (95% CI 0.1–18.5); P  < 0.001] and a direct correlation between Lp(a) and AER was observed ( r  = 0.34; P  < 0.01). AER was significantly different when Lp(a) phenotypes were considered ['small': median 19 μg min⁻¹ (range 1–195); 'big': median 9.5 μg min⁻¹ (range 1–186); 'null': 4 μg min⁻¹ (range 1–9); P  = 0.04]. None of the NIDDM patients with a 'null' phenotype showed an AER of > 10 μg min⁻¹. In conclusion, this case–control study provides evidence that microalbuminuria is associated with high serum Lp(a) in NIDDM without clinically detectable macroangiopathy. Furthermore, NIDDM patients with a 'null' phenotype could be considered at low risk for the development of microalbuminuria.     

p19ARF和p16INK4a在人胰腺癌、结直肠癌中表达的比较研究

目的：比较p16^INK4a和p19^ARF2种抑癌基因在可切除的人胰腺癌和结直肠癌组织中的表达形式，从而对其在这两种肿瘤的发生、发展中的意义及两种肿瘤生物学行为差异的本质有一个初步认识。方法：胰腺癌和结直肠癌切除标本各30例的石蜡标本，用单克隆抗体免疫组化ABC法同时测定肿瘤组织的p16、p19和p21蛋白的表达。结果：胰腺癌的p16表达率13.3％(4/30)明显低于结直肠癌的40％(12/30，Pd0.02)；而p19在两者的表达率差异不明显。p16与p19表达无相关性。p16的表达率与胰腺癌的临床病理分期和分级呈负相关；有大血管浸润的胰腺癌p19阳性率(20％)低于无血管浸润者(72.2％,P=0.01)。P19在局部淋巴结阳性的胰腺癌表达率低于淋巴结阴性者，但无显著差异。而p16和p19与结直肠癌的病理分期、分级、血管浸润和淋巴结转移均无明显相关。结论：p16的下调是胰腺癌发生发展的一个重要原因，可能也是胰腺癌生物学行为明显不同于结直肠癌的主要原因；我们还首次发现p19下调与胰腺癌的大血管浸润和淋巴结转移有关，p19异常可能是胰腺癌播散的1种标示。未发现这2种抑癌基因与结直肠癌发生发展的明显关系。     

南充市Rh阴性献血者血清学表型和不规则抗体调查

目的调查南充市Rh阴性无偿献血者的血清学表型和不规则抗体情况。方法应用血型血清学方法对初检Rh阴性的献血者进行Rh阴性确认、血清学表型鉴定及不规则抗体筛选鉴定。结果在433份送检标本中共确认Rh阴性402份(92.8%),检出D变异体23份(5.3%),其余8份(1.9%)为检验科RhD检测错误;共检出不规则抗体12份,在Rh确认阴性献血者中检出7份抗-D(1.7%);Rh阴性献血者的血清学表型分布结果主要以dccee和dCcee为主,其余为dCCee、dccEe和dCcEe。结论通过本次调查,掌握了南充市Rh阴性无偿献血者的血清学表型和不规则抗体的分布情况,保障了临床Rh阴性患者输血安全性、有效性和及时性。     

Genetic and environmental determinants of the PON-1 phenotype

BACKGROUND: Paraoxonase (PON-1) is a high-density lipoprotein (HDL)-associated enzyme that may protect against cardiovascular disease (CVD), because it hydrolyses oxidized phospholipids of low-density lipoprotein (LDL) and therefore prevents the detrimental effects on the arterial wall. The current report describes the determinants of PON-1 bioavailability and activity. MATERIALS AND METHODS: This is the largest (n = 1527) cross-sectional evaluation performed on PON-1 genotypes (Q192R, T-107C and L55M) and environmental determinants to PON-1 catalytic activity and bioavailability in serum of postmenopausal women. PON-1 catalytic activity and PON-1 bioavailability were measured, in vitro, with a paraoxon hydrolysis assay and a phenylacetate hydrolysis assay, respectively. RESULTS: The major determinant of paraoxon hydrolytic activity is the Q192R genotype, but there was also a relation with the C-107T and L55M genotype, HDL levels and alcohol consumption. Phenylacetate hydrolytic activity was most strongly affected by the C-107T genotype followed by the L55M genotype, HDL levels, alcohol consumption and smoking. CONCLUSIONS: PON-1 Q192R, C-107T and L55M genotype, alcohol consumption, smoking and HDL levels are determinants of serum PON-1 phenotype. The contributions of the genetic markers to the PON-1 phenotype are stronger than the contributions of the lifestyle determinants.     

pEGFP-BMI-1真核表达载体的构建、鉴定及其表达

本研究构建真核表达载体pEGFP-BMI-1并观察其在HeLa细胞中的表达。将BMI-1逆转录聚合酶链反应(RT-PCR)产物克隆入pEGFP-N1,经酶切、PCR鉴定及测序分析,构建pEGFP-BMI-1真核表达载体;采用脂质体转染法,将融合基因pEGFP-BMI-1转入HeLa细胞,用荧光显微镜和Western blot检测其表达,SYBR Green I实时定量RT-PCR方法检测转染前后HeLa细胞中P16INK4a mRNA的表达变化。结果表明:重组后的pEGFP-N1质粒已成功载入BMI-1的全长编码基因,序列测定的结果与预期设计完全一致。荧光显微镜下可见在转染pEGFP-BMI-1的HeLa细胞中存在荧光分布;Western blot检测发现存在外源性融合蛋白BMI-1-EGFP的表达。在HeLa细胞中过表达BMI-1显著下调P16INK4a mRNA的表达为对照组的9.2%(P<0.01)。结论:成功构建了真核表达质粒pEGFPBMI-1,转染HeLa细胞后可表达融合蛋白BMI-1-EGFP。这为今后研究BMI-1在肿瘤发生发展中的机制奠定了基础。     

Decrease in lipoprotein(a) after renal transplantation is related to the glucocorticoid dose

Abstract. Serum lipoprotein(a) [Lp(a)] concentrations and apolipoprotein(a) phenotypes were determined in 46 patients with end-stage renal disease both before as well as 1 week and 1, 3 and 6 months after renal transplantation. Immunosuppressive therapy consisted of cyclosporin A, prednisone and azathioprine. Before transplantation median Lp(a) levels did not differ between the patients and a healthy control group. A highly significant decrease ( P <0.001) in Lp(a) levels was observed in both male and female patients 1 week after transplantation. This marked reduction in Lp(a) occurred at a time when patients were receiving the highest doses of corticosteroids. As steroid doses were gradually tapered. Lp(a) concentrations subsequently increased, although at 6 months levels were still significantly reduced ( P <0.01) in women. No significant correlation was observed between Lp(a) and whole-blood cyclosporin levels, nor was there any correlation with the azathioprine dose. The reduction in Lp(a) concentrations was seen for all apo(a) phenotypes observed in the study.