结合珠蛋白遗传性多态现象与疾病关系的研究——Ⅰ.广东群体结合珠蛋白遗传性多态现象的特点

本文测定了广东地区1,083名无关中国人个体的血清Hp表型。观察到在这一人群中的Hp表型,频率分别为:Hp 2-2 0.500,Hp2-1 0.406,Hp1-1 0.088;Hp基因频率分别为Hp~2 0.703,Hp~1 0.291;均处于Hardy-Weinberg平衡状态。观察到Hp~1基因频率随年龄增加而降低。此外,在这一群体中还观寮到少见的Hp0型,其频率为0.006。对其中4例作了家谱调查,结果支持可能存在Hp~0基因的假设,推测其遗传方式可能与ABO血型相类似。     

广西侗、瑶、壮、汉族结合珠蛋白的遗传多态性

本文报道了广西侗族、瑶族、壮族和汉族共554例受检者的Hp表型分布和基因频率。经χ2检验,4个群体中Hp表型分布符合Hardy-Weinberg遗传平衡法则。比较4个群体间Hp基因频率,无显著性差异,但汉族Hp1-1表型频率(0.110)显著高于侗族(0.0156)。本研究检出2-1M、2-H、2-1Tran、1-H和2-1H aw等5种变异型。 Abstract:This article deals with the phenotypes and gene frequencies of haptoglobin from 554 subjects belonging to 4 different ethnics,Dong,Yao,Zhuang and Han in Guangxi.The distribution of Hp phenotype in 4 ethnic groups is in agreement with the Hardy-Weinberg law.The gene frequencies of haptoglobin were compared with each other and no evidence of difference among these 4 populations was found.The variants,including 2-1M,2-H,2-1Tran,1-H and 2-1Haw were discovered in the experiment.     

白皮松的保育遗传学研究I．基因保护分析

在白皮松主要分布区抽取了10个天然群体进行酶电泳分析,共测定了16种酶系统,得到31个清晰酶位点和53个等位基因。按照基因频率进行分类,53个基因中有32个全局基因、14个广域基因、6个局域基因和1个特异基因。通过计算机模拟建立基因捕获曲线的结果表明,随机抽取5个群体可平均捕获99．9％的基因。群体的等位基因频率与地理生态因子的相关分析表明,Idh与Pgi—2两个位点的基因频率呈现明显的梯度变异。该项研究为有效保护白皮松天然群体的基因资源提供了依据。     

不同地区粘虫群体的同工酶变异

本文用聚丙烯酰胺凝胶电泳测定了粘虫(Mythirana separata)五个地区群体(福建、贵州、安微、北京和吉林)的同工酶变异.分析10种同工酶(ADH,AO,GOT,α-GPDH,HK,LAP,LDH,MDH,ME和TYR)的14个座位,其中单型座位有8个,多型座位有6个,后者占42.9%.五个地区群体的平均杂合度为0.138.根据14个基因座位的等位基因频率计算出的五个地区群体间的平均遗传相似度为0.978,平均遗传距离为0.023,表明这五个地区群体的基因结构基本相似,遗传分化程度很小.可以认为,在这五个地区群体之间不存在基因流动的障碍.这从遗传和变异的角度证明了粘虫的远距离迁飞特性,迁飞阻止了不同地区群体的遗传分化.     

不同地区粘虫群体的同工酶变异

本文用聚丙烯酰胺凝胶电泳测定了粘虫(Mythirana separata)五个地区群体(福建、贵州、安微、北京和吉林)的同工酶变异.分析10种同工酶(ADH,AO,GOT,α-GPDH,HK,LAP,LDH,MDH,ME和TYR)的14个座位,其中单型座位有8个,多型座位有6个,后者占42.9%.五个地区群体的平均杂合度为0.138.根据14个基因座位的等位基因频率计算出的五个地区群体间的平均遗传相似度为0.978,平均遗传距离为0.023,表明这五个地区群体的基因结构基本相似,遗传分化程度很小.可以认为,在这五个地区群体之间不存在基因流动的障碍.这从遗传和变异的角度证明了粘虫的远距离迁飞特性,迁飞阻止了不同地区群体的遗传分化.     

白皮松的保育遗传学研究Ⅰ.基因保护分析

在白皮松主要分布区抽取了10个天然群体进行酶电泳分析,共测定了16种酶系统,得到31个清晰酶位点和53个等位基因。按照基因频率进行分类,53个基因中有32个全局基因、14个广域基因、6个局域基因和1个特异基因。通过计算机模拟建立基因捕获曲线的结果表明,随机抽取5个群体可平均捕获99.9%的基因。群体的等位基因频率与地理生态因子的相关分析表明,Idh与Pgi^-2两个位点的基因频率呈现明显的梯度变异。该项研究为有效保护白皮松天然群体的基因资源提供了依据。     

不同群体中ATXN2基因编码区CAG重复的变异研究

在中国6个生活环境差异较大的少数民族群体中进行ATXN2基因编码区CAG重复的变异研究,以衡量其是否受到正选择的作用以及寻找推动选择作用的因素。采集6个民族群体共291个健康无关个体,对其进行STR分型,直接计数其等位基因及等位基因型频率,计算其线性Fst值,构建针对该基因的系统进化树,并对各群体进行MDS分析。线性Fst值结果显示:回族和彝族群体间ATXN2基因STR位点进化的差异具有显著性,其他4个群体相互间无显著性差异。结合已报道的其他群体进一步分析,回族、哈尼族、云南蒙古族以及内蒙古自治区蒙古族每个人群都与日本人群有显著性差异;回族、内蒙古自治区蒙古族与汉族具有显著性差异。6个群体中ATXN2基因STR的等位基因频率有各自的分布特点,稀有等位基因频率变化产生的原因可能是选择作用的结果。     

结合珠蛋白遗传性多态现象与疾病关系的研究——Ⅱ.结合珠蛋白遗传性多态现象与白血病的关系

本文报告广东地区0—54岁白血病病人Hp~1频率显著高于正常人,其相对危险率为1.9,提示这一地区Hp~1基因与白血病发生有关。急粒和急淋组Hp~1频率显著高于正常人,慢粒组则与正常人无显著差异。化疗后的急拉和急淋组Hp2-2型和Hp2-1型Hp平均含量较化疗前显著降低,Hp1-1型则降低不显著;慢粒组三类Hp平均含量化疗前后无明显变化。提示Hp~1基因可能与急性白血病发生有关。具有Hp纯合子或杂合子正常母亲的白血病子女中,纯合子与杂合子的比例符合预期比例,提示在白血病病人家庭中Hp的遗传方式无异常。还观察到,肝癌中Hp~1频率亦显著增高,提示Hp~1基因除与白血病有关外,还可能与其他某些肿瘤的发生有关。     

利用微卫星技术分析不同类群鸡的遗传多样性

利用8个微卫星标记分析了6个生产类群鸡的遗传多样性。计算了各群体在各位点上的等位基因频率,并据此计算出各群体的平均遗传杂合度(h)、多态信息含量(PIC)和有效等位基因数(Ne)。结果表明:6个鸡群在8个微卫星座位上的基因频率存在明显的差异,其平均基因杂合度为0.7317,平均多态信息含量为0.6815。其中,群体平均杂合度最高的是安卡红鸡,为0.7716;平均杂合度最低的是新罗曼鸡,为0.7073。所选的8个微卫星座位均为高度多态,可作为有效的遗传标记用于鸡群体遗传多样性的分析。     

迪庆藏猪的遗传多样性研究

采用水平板淀粉凝胶电泳法检测了52头迪庆藏猪的13个血液蛋白座位的多态性,并计算各座位等位基因的频率及其估计误差以及基因频率估计值的精确度和可靠性。结果发现,迪庆藏猪在Tf、Cp、Am、Hp、6PGD、CEs、Ca共7个座位表现出多态性,并分别受控于3、2、5、5、2、2、3个常染色体等位基因。其余6个座位(Pa 、EsD、 PHI、 PGM、 G6PD、 MDH)未检测出多态性。     

Microheterogeneity of mammalian haptoglobins in isoelectric focusing.

1. Human haptoglobin type 1-1, porcine haptoglobin, and equine haptoglobin were isolated and purified. 2. These haptoglobins were similar in polyacrylamide gel electrophoresis and in subunit structure but showed microheterogeneity in isoelectric focusing. 3. Isoelectric points of human haptoglobin as determined with photopolymerized gels were found to be 4.03-4.24, of porcine haptoglobin 4.0-4.30, and of horse haptoglobin 3.80-4.15, respectively. 4. Results obtained with chemically polymerized gels were 0.08-0.3 pH units higher. 5. Examined haptoglobins differed also in the ability of complex formation with hemoglobin, in sialic acid content and in antigenic specificity.     

Effect of modification on physicochemical and biological properties of haptoglobin. VI. Reaction with azlactone of p-nitrobenzoyl-valine.

The azlactone of p-nitrobenzoyl-valine (Nbz-Val) has been used for modification of xi-amino groups of lysine in haptoglobin type 1-1, in hemoglobin, and in the haptoglobin-hemoglobin complex. By the use of this reagent 95% of amino groups in haptoglobin and 90% in hemoglobin have been blocked without any changes in peroxidase activity of the formed complexes: Nbz-Val.haptoglobin with hemoglobin, Nbz-Val. hemoglobin with haptoglobin, and Nbz-Val.(haptoglonin-hemoglobin). After reduction and reoxidation, Nbz-Val.haptoglobin was found to retain 90% of peroxidase activity when complexed with hemoglobin. Beta chains separated either from haptoglobin or Nbz-Val.haptoglobin showed 15% of peroxidase activity in the complex with hemoglobin, alpha chains of the same origin were completely inactive. Whereas recombination of haptoglobin from alpha and beta chains resulted in 42% hemoglobin-binding capacity, renaturation of Nbz-Val.haptoglobin from separated subunits was found to proceed with almost 100% yield. In immunodiffusion with rabbit anti-haptoglobin or anti-Nbz-Val.haptoglobin sera, preparations of haptoglobin and Nbz-Val.haptoglobin after reduction and reoxidation or after recombination from separated subunits gave similar precipitation arcs showing the reaction of immunological identity.     

Haptoglobin binding to apolipoprotein A-I prevents damage from hydroxyl radicals on its stimulatory activity of the enzyme lecithin-cholesterol acyl-transferase

Apolipoprotein A-I (ApoA-I), a major component of HDL, binds haptoglobin, a plasma protein transporting to liver or macrophages free Hb for preventing hydroxyl radical production. This work aimed to assess whether haptoglobin protects ApoA-I against this radical. Human ApoA-I structure, as analyzed by electrophoresis and MS, was found severely altered by hydroxyl radicals in vitro. Lower alteration of ApoA-I was found when HDL was oxidized in the presence of haptoglobin. ApoA-I oxidation was limited also when the complex of haptoglobin with both high-density lipoprotein and Hb, immobilized on resin beads, was exposed to hydroxyl radicals. ApoA-I function to stimulate cholesterol esterification was assayed in vitro by using ApoA-I-containing liposomes. Decreased stimulation was observed when liposomes oxidized without haptoglobin were used. Conversely, after oxidative stress in the presence of haptoglobin (0.5 microM monomer), the liposome activity did not change. Plasma of carrageenan-treated mice was analyzed by ELISA for the levels of haptoglobin and ApoA-I, and used to isolate HDL for MS analysis. Hydroxyproline-containing fragments of ApoA-I were found associated with low levels of haptoglobin (18 microM monomer), whereas they were not detected when the haptoglobin level increased (34-70 microM monomer). Therefore haptoglobin, when circulating at enhanced levels with free Hb during the acute phase of inflammation, might protect ApoA-I structure and function against hydroxyl radicals.     

Haptoglobin study of sheep during the development process]

The development of haptoglobin blood system was studied in the sheeps during intrauterine development and early postnatal period. The haptoglobin content was shown to decrease with the foetus age, two peaks of its reliable increase having been, however, noted -- on the 55th and 105th days of development. After the birth the haptoglobin concentration in blood is relatively low, increases gradually and attains by the 8th month of life that in adult animals. In the blood serum of 45--120 days old foetuses two phenotypes of fetal haptoglobin were found; the adult haptoglobin is present only beginning from the 1st month of life.     

Role of Haptoglobin in Polycystic Ovary Syndrome (PCOS), Obesity and Disorders of Glucose Tolerance in Premenopausal Women

Background

Hp² alleles of the haptoglobin α–chain polymorphism reduce the anti-oxidant properties and increase the pro-inflammatory actions of this acute-phase protein in a gene-dosage fashion. We hypothesized that the haptoglobin polymorphism might contribute to the increased oxidative stress and low-grade chronic inflammation frequently associated with polycystic ovary syndrome, obesity, and abnormalities of glucose tolerance.

Methodology/Principal Findings

Serum haptoglobin and the haptoglobin α–chain polymorphism were determined in 141 patients with polycystic ovary syndrome and 102 non-hyperandrogenic women. Of the whole group of 243 premenopausal women, 117 were obese and 51 showed abnormal glucose tolerance. Although serum haptoglobin concentrations were similar in PCOS patients and controls, the former presented with an increased frequency of Hp² alleles (62% vs. 52%, P = 0.023). Circulating haptoglobin levels increased with obesity (P<0.001), yet no association was found between obesity and haptoglobin genotypes. No differences were observed in haptoglobin levels or genotype frequencies depending on glucose tolerance. Fifty percent of the variation in serum haptoglobin concentrations was explained by the variability in serum C-reactive protein concentrations, BMI, insulin sensitivity and haptoglobin genotypes.

Conclusions/Significance

Serum haptoglobin concentrations in premenopausal women are largely dependent on the haptoglobin polymorphism and on the presence of obesity, with insulin resistance and chronic inflammation possibly modulating this relationship. The association of polycystic ovary syndrome with Hp² alleles suggests that the anti-oxidant and anti-inflammatory properties of haptoglobin may be reduced in these patients.     

Hydrazinoacridine staining of proteins and glycoproteins in polyacrylamide gels

The histochemical method for staining polyaldehydes in tissue sections with p-hydrazinoacridine has been adapted for use in polyacrylamide gels. While staining of histological preparations was reported to be specific for polyaldehydes and independent of bisulfite, both glycoproteins (β chain of haptoglobin) and nonglycoproteins (lysozyme and α chain of haptoglobin) were stained following periodate oxidation, and satisfactory results were highly dependent on the presence of bisulfite. Hydrazinoacridine staining of periodate-treated gels produced an extremely sensitive fluorescent labeling of the haptoglobin β chain and also stained haptoglobin α chain and lysozyme. The proteins could be visualized under visible light as yellow bands which were scanned spectrophotometrically at 440 nm. The β chain of haptoglobin could be subjectively distinguished from the nonglycoproteins both by differential intensity of staining with hydrazinoacridine and Coomassie brilliant blue and by the yellow nature of the fluorescence. The sensitivity of hydrazinoacridine staining of the β chain of haptoglobin compared favorably to that of the commonly used periodic acid-Schiff staining procedures and provided the advantage that nonglycoproteins in complex mixtures could be localized in the gels.     

Effect of dystocia on serum haptoglobin in Awassi ewes

Serum haptoglobin concentration was determined in 102 Iraqi Awassi ewes. Blood samples were collected from 82 ewes before the correction of dystocia, 10 ewes with eutocia 2 to 4 h after parturition and 10 nonpregnant ewes during the seasonal anestrus phase. The mean serum haptoglobin concentration was significantly higher (P < 0.01) in ewes with dystocia than in ewes with normal births and in the nonpregnant ewes. No significant difference was found between serum haptoglobin concentrations of ewes with ringwomb and ewes with dystocia due to other causes. There was a significant elevation (P < 0.01) of serum haptoglobin in cases treated 24 h after labor compared with those treated during the first 24 h. Significant (P < 0.05) differences were found between serum haptoglobin concentrations in ewes treated surgically and those treated manually.     

Calorimetric studies of the haemoglobin–haptoglobin reaction

Haptoglobin binds haemoglobin so firmly that there is practically no dissociation. It would be expected that the heat of the reaction would be relatively large. The development of the microcalorimeter by Benzinger offered the opportunity to measure the heat of reaction. The experiments were carried out in the Beckman 190B Microcalorimeter in two ways: (1) a constant amount of haptoglobin (Kabi; 65mg.) with different amounts of haemoglobin, and (2) a constant amount of haemoglobin (32.5mg.) with different amounts of haptoglobin. The proteins, each in 5ml. of 0.15m-phosphate buffer, pH7.4, were placed in equal-volume calorimeter cells. The heat produced/mg. of haemoglobin was calculated from the slope of the curve for a constant amount of haptoglobin and from the maximum heat for a constant amount of haemoglobin. This heat is about 70kcal./mole at 37 degrees . DeltaH varies with temperature, being -70.2 at 37 degrees , -29.7 at 20 degrees and 7.2 at 4 degrees . From the amount of haptoglobin required to attain maximum heat with 32.5mg. of haemoglobin and the amount of haemoglobin required to attain maximum heat with 65mg. of haptoglobin, it appears that at excess of haptoglobin there is competition between the reactions of 2moles of haptoglobin with 1mole of haemoglobin (or 2 alphabeta-chains) and 1mole of haptoglobin with 1mole of haemoglobin.     

Regulation of haptoglobin gene expression in 3T3-L1 adipocytes by cytokines, catecholamines, and PPARgamma

Factors which regulate expression of the haptoglobin (acute phase reactant) gene in adipocytes have been examined using 3T3-L1 cells. Haptoglobin expression was observed by Northern blotting in each of the major white adipose tissue depots of mice (epididymal, subcutaneous, mesenteric, and perirenal) and in interscapular brown fat. Expression occurred in mature adipocytes, but not in the stromal-vascular fraction. In 3T3-L1 cells, haptoglobin mRNA was detected from day 4 after the induction of differentiation into adipocytes. Lipopolysaccharide and the cytokines, TNFalpha and interleukin-6, resulted in substantial increases in haptoglobin mRNA in 3T3-L1 adipocytes; the increase (7-fold) was highest with TNFalpha. Increases in haptoglobin mRNA level were also induced by dexamethasone, noradrenaline, isoprenaline, and a beta3-adrenoceptor agonist. In contrast, haptoglobin mRNA was reduced by nicotinic acid and the PPARgamma agonist, rosiglitazone. RT-PCR showed that the haptoglobin gene was expressed in human adipose tissue (subcutaneous, omental). It is concluded that haptoglobin gene expression in adipocytes is stimulated by inflammatory cytokines, glucocorticoids, and the sympathetic system, while activation of the PPARgamma nuclear receptor is strongly inhibitory.