红细胞补体受体1(CR1)数量基因多态性与反复呼吸道感染易感性的研究

目的通过研究红细胞补体受体1(CR1)数量基因多态性与反复呼吸道感染的相关性,探讨反复呼吸道感染(RRTI)患儿的遗传易感因素.方法利用限制性内切酶HindⅢ,聚合酶链反应限制性片段长度多态性(PCR-RFLP)分析,基因测序等技术测定58例RRTI患儿(病例组)和56例正常儿童(对照组)的红细胞CR1数量基因多态性,并进行比较.结果病例组中CR1基因HH、HL和LL基因型分布频率分别为34.2%、55.3%和10.5%,而对照组中HH、HL和LL基因型分布频率分别为75%、21.4%和3.6%.两组CR1基因型的分布频率差异有显著性(P＜0.001).病例组中HL和LL基因型占优势(OR=5.77).两组CR1基因等位基因的分布频率差异也有显著性(P＜0.01),病例组中L等位基因分布频率高于对照组.结论红细胞补体受体1(CR1)数量基因多态性与反复呼吸道感染有相关性,提示CR1基因HindⅢ酶切位点多态性可能在决定个体反复呼吸道感染遗传易感性方面有重要作用.     

正常人红细胞CR1密度相关基因组多态性分布分析

采用PCR和Hind Ⅲ酶切技术研究了正常人红细胞CR1密度相关基因组多态性,发现在189例中国正常人群中,红细胞CR1密度相关基因HH型比率是79％,HL型比率是18％,LL型比率是3％。在30岁以上成年人中,64例男性的HH型（85.9％）明显高于63例女性HH型比率（68.2％,P<0.01）。在94例女性组中,60岁以上人群的HH型比率是62％与10～19岁女性人群的HH型比率94％相比有显著差异（P<0.05）。这些结果表明老年人红细胞CR1密度相关基因多态性与性别相关。     

Ⅱ型糖尿病患者红细胞天然免疫功能变化与CR1密度基因多态性的相关分析

目的　研究Ⅱ型糖尿病 (ⅡDM)患者的红细胞CR1密度基因多态性及红细胞免疫粘附功能 ,探讨ⅡDM患者红细胞免疫粘附功能低下的原因。方法　分ⅡDM组和健康对照组 ,用PCR RELP方法测红细胞CR1密度基因多态性 ,红细胞天然免疫粘附活性用红细胞C3b受体花环率(EC3bRR)及免疫复合物花环率 (ECICR)测定。结果　 (1)ECR1基因构成 ,ⅡDM组HH型为 5 7.14 %、HL型 37.5 0 %、LL型 5 .36 % ;对照组HH型为 6 8.86 %、HL型 2 8.5 7%、LL型 3.5 7% ;ⅡDM组的基因缺陷率为 4 2 .86 % ,对照组为 32 .14 %。两组相比基因构成及缺陷率差异均无显著性 (P >0 .0 5 )。 (2 )与对照组相比 ,ⅡDM组EC3bRR下降 ,ECICR上升 (P <0 .0 1)。 (3)两组组内比较 ,HH型的EC3bRR均明显高于HL型 ,ECICR均低于HL型。结论　ⅡDM患者红细胞CR1密度基因HH型比率下降不显著 ,但红细胞天然免疫功能低下程度与ECR1基因缺陷关系密切相关。不同ECR1基因型的人群红细胞免疫功能有差异 ,HH型的红细胞免疫粘附功能强于HL型。     

卵巢癌患者红细胞CR1密度相关基因组多态性与血细胞天然免疫活性相关性研究

目的　对卵巢癌患者血细胞天然免疫活性与红细胞CR1密度相关基因组多态性的相关性进行对比研究。方法　对 5 1例卵巢癌、4 3例良性肿瘤患者和 4 0例正常妇女采用PCR RFLP方法测定红细胞CR1(ECR1)密度相关基因多态性 ,采用红细胞或淋巴细胞免疫粘附肿瘤细胞的方法对血细胞天然免疫活性进行了测定。结果　发现卵巢癌患者红细胞和淋巴细胞CR1天然免疫活性明显低于良性卵巢肿瘤患者和正常人。红细胞CR1密度相关基因高表达 (HH)型组的血细胞天然免疫活性明显高于中表达 (HL)型 ,而HL型明显高于低表达 (LL)型。加红细胞组淋巴细胞CR1天然免疫活性明显高于不加红细胞组 (P <0 .0 1)。结论　红细胞增强淋巴细胞CR1天然免疫活性与红细胞CR1密度相关基因型密切相关。     

HIF1A基因两个位点多态性与夏尔巴人群高原低氧适应关系的研究

目的研究西藏高原夏尔巴人群缺氧诱导因子1α（hypoxia-inducible factor 1，alpha subunit；HIF1A）基因（HIF1A）第12外显子1772（C→T）、1790（G→A）多态性与高原低氧适应相关性。方法选取世居西藏高原夏尔巴族148人及广东汉族健康个体90人的血样，提取白细胞基因组DNA，聚合酶链反应-限制性片段长度多态性（polymerase chain reaction-restriction fragment length polymorphism，PCR-RFLP）检测HIF1A基因第12外显子1772（C→T）、1790（G→A）的单核苷酸多态性，分析其基因多态性特征。结果1772（C→T）CC、CT和TT基因型频率在夏尔巴人组与汉族对照组分别为14．19％和16．67％、39．19％和41．11％、46．62％和42．22％，两组间比较差异无统计学意义；1790（G→A）GG、GA和AA基因型频率在夏尔巴人组与汉族对照组分别为57．43％和75．56％、37．84％和21．11％、4．73％和3．33％，夏尔巴人组的GG基因型频率较汉族对照组的低（P〈0．01），而GA基因型频率高于汉族对照组（P〈0．01）。组合基因型分布，夏尔巴人CC＋GA、CT＋AA、TT＋GA和TT＋AA的组合基因型频率高于汉族组。结论HIFIA基因1790（G—A）多态性与夏尔巴人群高原低氧适应存在相关性，GA、AA基因型可能对低氧适应有利，值得进一步深入探讨。     

中国北方汉族人细胞色素P450 1A1基因MspⅠ多态性与早发性帕金森病关系的研究

目的：探讨中国北方汉族人细胞色素P4501A1基因MspI多态性与早发性帕金森病的易感性的关系。方法：用聚合酶链反应－限制性片段长度多态性技术,分析了126例早发性帕金森病患者（发病年龄＜50岁）和172名正常健康成人CYP1A1基因3’端限制性内切酶MspI位点的3种基因型（A、B、C）的分布频率。结果：MspI基因型C在病例组和对照组中各占15．1％和13．4％,基因型A在两组中分别占41．3％和34．9％,基因型B在两组中分别占43．6％和51．7％,各基因型在两组中相比较,差异无显著性（P＞0．05）。C基因型在病例组和对照组分别占36．9％和39．2％,两者差异无显著性（P＞0．05）。结论：提示解毒酶CYP1A1基因MspI多态性的单独存在可能与早发性帕金森病的易患性无关。     

中山地区人群KCNQ1基因多态性与妊娠期糖尿病相关性研究

目的探讨电压门控性钾通道Q亚家族成员1（KCNQl）基因rs2237892（C／T）,m2237895（A／C）,rs2237896（AVG）位点单核苷酸多态性与中山地区人群妊娠期糖尿病（GDM）的关系。方法本研究共采用2011年-2013年中山市南朗医院和中山市博爱医院共285例孕妇,其中GDM组185例,血糖正常100例设为对照组。采用聚合酶链反应-限制性片段长度多态性（PCR—RFLP）检测KCNQl基因多态性,评估其与GDM的相关性。GDM组和对照组基因型分布运用logistic回归模型分析。GDM组相关表型运用多重回归模型分析。结果（1）SNPrs2237896的三种基因型（AA、AG、GG）在GDM组和对照组分布频率分别是4．8％,42．2％,53．0％和3．0％,28．0％,69．0％,两组的基因型分布频率差异显著（P=0．032,P〈0．05）。Rs2237896等位基因A、G的分布频率（26％。74％）高于对照组（17％,83％）,差异具有显著性（P=0．015）。（2）SNPm223782中,其三种基因型（CC、CT、1Tr）在GDM组和对照组分布频率分别是24．8％,68．1％,8．1％和38％,49％,13％,其基因型分布频率两组具有显著差异（P=0．007,P〈0．05）,但是Rs2237892等位基因C、T的分布频率并不具有差异（P=0．279,P〉0．05）。结论KCNQ1基因SNPrs2237896多态性可能与中国中山市人群中GDM发病具有一定相关性。     

RANTES基因-28C／G单核苷酸多态性与子宫内膜异位症遗传易感性的研究

目的探讨调节正常T细胞表达和分泌活性因子（regulated on activation, normal T cell expressed and secreted,RANTES）基因启动子区-28C／G单核苷酸的多态性与广东籍汉族患者子宫内膜异位症的关系。方法应用聚合酶链反应-限制性酶切片段长度多态性技术（PCR—RFLP）并进行基因测序的方法检测广东籍汉族子宫内膜异位症患者59例（内异症组）,非子宫内膜异位症患者49例（对照组）,比较分析各组间基因型频率和等位基因频率。结果RANTES基因启动子区-28C／C基因型在子宫内膜异症组及对照组分布频率分别为81．36％、81．63％,C／G基因型分布频率分别为18．64％、18．37％;两组间的基因型分布频率比较差异无显著性（P〉0．05）。RANTES基因启动子区-28位点C等位基因型在内异症组及对照组中的分布频率分别为90．68％、90．82％,G等位基因型分布频率分别为9．32％、9．18％,两组间等位基因型频率比较差异无统计学意义（P〉0．05）。结论在广东籍汉族妇女中,RANTES基因启动子区-28C／G单核苷酸多态性与子宫内膜异位症遗传易感性可能无关联。     

甘露糖结合凝集素与儿童常见感染易感性的关系研究

目的 研究浙江省汉族儿童中甘露糖结合凝集素(MBL)基因多态性和蛋白水平与常见感染(反复呼吸道感染、急性呼吸道感染、CMV活动性感染、体表局部脓肿、中耳炎)易感性的关系.方法 用PCR和测序法对感染组和对照组儿童的MBL基因启动子区和外显子1区的6个突变位点进行检测和分型,用ELISA试剂盒检测两组儿童的血浆MBL蛋白浓度.结果 感染组和对照组均未检测到MBL基因外显子1区+223位点(C/T)和+239位点(G/A)的突变,对照组也未检测到启动子区+4位点(C/T)的突变.启动子区-550位点3种基因型HH、HL、LL在感染组和对照组的频率分别为43.1％、29.1％、27.8％和63.8％、17.1％、19.1％,两组间基因型频率差异有统计学意义(P＜0.05).启动子区和外显子1区所有基因型组合形成的完整基因型可分为与血浆蛋白浓度相关的“YA型”和“XB型”,两种完整基因型频率在感染组和对照组间差异有统计学意义(P＜0.05).感染组和对照组血浆MBL蛋白浓度均呈偏态分布.CMV感染组蛋白浓度低于对照组,急性呼吸道感染和局部脓肿组蛋白浓度高于对照组,差异均有统计学意义(P＜0.05).结论 包括启动子区和外显子1区在内的MBL基因多态性与儿童常见感染的易感性具有一定相关性.     

雌激素受体1基因多态性与重症肌无力关系的初步研究

目的 探讨雌激素受体-1基因多态性在MG患者中的分布特征及其与免疫治疗疗效的关系。方法 选取72例MG患者和50例青年卒中患者（除外自身免疫病）的全血，提取DNA后使用序列特异性PCR扩增技术（SSP-PCR）测定雌激素受体-1基因多态性的表达，比较基因型和等位基因在各种MG特征组（包括性别、年龄、病型、合并胸腺异常、激素疗效）的分布。结果 密码子10基因型分布频率在激素有效组和无效组的基因型分布频率有显著性差异（P＝0.043），激素无效组与对照组的基因型分布频率有显著性差异（P＝0.019），考虑到受累范围的影响，基因型的差别在激素疗效各组没有显著性（P＝0.096）。在其他亚组间及各亚组与对照组间均无显著性差异。密码子10等位基因分布频率在各亚组间均无显著性差异（P>0.05）。密码子87在实验组、对照组均为GCG型纯合子，不具有多态性。结论 ESR-1基因密码子10、87基因型分布频率对MG发病及临床特征无显著影响。     

Bilirubin-induced erythrocyte membrane cytotoxicity

The kinetics of bilirubin erythrocyte interaction have been followed by scanning electron microscopy. Bilirubin-induced erythrocyte cytotoxicity embodies the interaction of the bile pigment with the outer half of the erythrocyte plasma membrane bilayer couple. This interaction leads to crenation. This membrane event appears to be primary and precedes hemolysis. The membrane crenation is dependent on the concentration of the bile pigment and is reversed by bovine serum albumin again in a concentration-dependent manner. Phospholipids do not alter bilirubin erythrocyte ineraction. Erythrocytes from jaundiced neonates show crenated surface structure in scanning electron microscopy. The crenation depends upon severity of jaundice in neonates. This suggests similarity between in vivo and in vitro mechanisms of cytotoxicity mediated by the bile pigment. Further, phototherapy reverses the process of membrane crenation. The in vivo photocatabolities isolated from urine of jaundiced neonates are nontoxic to erythrocyte membrane.     

Determination of erythrocyte sorbitol

An increase in the intra-cellular concentrations of sorbitol can be responsible, at least in part, for certain long term complications of diabetes. Since the erythrocyte concentration of this polyol is a good indicator of that of other cells, we propose a simple, rapid enzymatic assay technique for red blood cells. The results already obtained reveal a significant difference between the erythrocyte sorbitol concentration in non-diabetic subjects and that in diabetic patients.     

Hazards due to erythrocyte alloantibodies

The hazardous effects of blood group alloantibodies with respect to immediate and delayed haemolytic transfusion reactions, the destruction of fetal red blood cells, and the damage of transplanted tissue (in kidney transplantation) are described.     

Suicidal erythrocyte death in sepsis

Sequelae of sepsis include anemia which presumably results from accelerated clearance of erythrocytes from circulating blood. The underlying mechanisms, however, remained hitherto elusive. Most recent studies disclosed that increased cytosolic Ca2+ activity and ceramide both trigger suicidal erythrocyte death (i.e., eryptosis), which is characterized by lipid scrambling of the cell membrane leading to phosphatidylserine exposure at the erythrocyte surface. Phosphatidylserine exposing erythrocytes may adhere to vascular walls or may be engulfed by macrophages equipped with phosphatidylserine receptors. To explore whether sepsis leads to eryptosis, erythrocytes from healthy volunteers were exposed to plasma of patients suffering from sepsis, or to supernatants from sepsis producing pathogens. Then, phosphatidylserine exposure (annexin V binding), cell volume (forward scatter), cytosolic Ca2+ activity (Fluo3 fluorescence), and ceramide formation (anti-ceramide antibody) were determined by flow cytometry. Challenge of erythrocytes with plasma from the patients but not with plasma from healthy individuals triggered annexin V binding. The effect of patient plasma on erythrocyte annexin V binding was paralleled by formation of ceramide and a significant increase of cytosolic Ca2+ activity. Exposure of erythrocytes to supernatant of pathogens similarly induced eryptosis, an effect correlating with sphingomyelinase activity. The present observations disclose a novel pathophysiological mechanism leading to anemia and derangement of microcirculation during sepsis. Exposure to plasma from septic patients triggers phosphatidylserine exposure leading to adherence to the vascular wall and clearance from circulating blood.     

Dielectric inspection of erythrocyte morphology

We performed a systematic study of the sensitivity of dielectric spectroscopy to erythrocyte morphology. Namely, rabbit erythrocytes of four different shapes were prepared by precisely controlling the pH of the suspending medium, and their complex permittivities over the frequency range from 0.1 to 110 MHz were measured and analyzed. Their quantitative analysis shows that the characteristic frequency and the broadening parameter of the dielectric relaxation of interfacial polarization are highly specific to the erythrocyte shape, while they are insensitive to the cell volume fraction. Therefore, these two dielectric parameters can be used to differentiate erythrocytes of different shapes, if dielectric spectroscopy is applied to flow-cytometric inspection of single blood cells. In addition, we revealed the applicability and limitations of the analytical theory of interfacial polarization to explain the experimental permittivities of non-spherical erythrocytes.