Anti-EnaFS detected in the serum of an MiVII homozygote

Anti-EnaFS was detected in the serum of a Caucasian woman, K.T., during her third pregnancy. She had not been transfused. At delivery, the infant's red cells (RBCs) had a negative direct antiglobulin test. The antibody was inhibited by isolated MN sialoglycoprotein (SGP). Unlike other En(a-) cells, K.T.'s EnaFS-negative RBCs were found to demonstrate normal sialic acid levels, and an MN SGP exhibiting normal sodium dodecylsulphate electrophoretic mobility and periodic acid-Schiff staining intensity. K.T., whose parents were consanguineous, appears to be the first known MiVII homozygote. In a three-generation family study, MiVII was shown to travel with MS. The Anek receptor of K.T.'s RBCs was found to be located on the trypsin-resistant and V8 protease-sensitive portion of the MN SGP (residues 40 to 57), whereas the Anek receptor of MiVI RBCs was shown to be associated with their abnormal Ss SGP. K.T.'s MN SGP appears to exhibit an amino acid sequence alteration in the region of the EnaFS determinant, which is located within the residues 46 to 56 of the normal MN SGP. These findings suggest that the MiVII and EnaFS receptors are the products of allelic genes.     

Studies on the blood of an MiV homozygote

An individual, whose parents are third cousins, has been shown to be homozygous for the rare Mi.V. condition. The proposita's red blood cells type as M−, N+(weak), S−, s+(strong), U+, Mi(a−), Vw−, Hil+; Wr(a− b−). The cells react, albeit less strongly than most other samples, with anti-Ena. However, from studies on the red blood cells of the proposita and on those of another person of the En(a+), Wr(a−b−) phenotype, it is apparent that the term "anti-Ena" actually describes a number of antibodies of differing specificities. Inhibition studies with sialoglycoprotein (SGP) isolates, and tests on protease-modified red blood cells illustrate some of the differences in specificity. Biochemical analyses of the SGPs of the red blood cells of the MiV homozygote and those of her parents confirm that the Mi.V condition is associated with the absence of normal MN SGP (alpha) and normal Ss SGP (delta), the appearance of a hybrid SGP molecule comprised of a portion of the MN SGP at its NH2 terminal end, and a portion of the Ss SGP at its C terminal end.     

中国回族首次检出苯丙酮尿症R176X突变型纯合子:1例报告

背景：苯丙酮尿症是由于苯丙氨酸羟化酶基因突变引起，苯丙氨酸羟化酶基因突变主要是因为碱基的置换，短片段、插入等。目的：鉴定回族苯丙酮尿症家系中苯丙氨酸羟化酶基因突变。设计：开放性实验。单位：解放军兰州军区乌鲁木齐总医院和北京首都儿科研究所。对象：患儿，男，回族，就诊时3岁1个月。1岁左右时发现智力滞后，近3岁时到医院就医，诊断为脑性瘫痪，反复治疗无效后于2004—12-13转入解放军兰州军区乌鲁木齐总医院就诊。尿液三氯化铁试验呈强阳性，血苯丙氨酸浓度测定1680μmol／L，遂确诊患儿为经典型苯丙酮尿症。方法：抽取患儿及父母静脉血各5mL，EDTA-Na2抗凝。基因组DNA提取采用经典的酚，氯仿法进行。PAH基因外显子7，6，11，3，12，5的PCR引物序列参照文献设计。PCR产物用2％琼脂糖凝胶电泳检测。取5μL PCR产物与等体积的变性缓冲液混匀，97℃变性5min，冰浴并迅速上样于80g／L非变性聚丙烯酰胺凝胶中电泳。电泳结束后常规方法银染，分析并记录单链DNA带型。采用PCR产物直接测序的方法，由上海博亚生物技术公司应用AB1377全自动序列分析仪（PE公司）完成样品纯化及序列分析。主要观察指标：尿液三氯化铁试验，血液血苯丙氨酸浓度和苯丙氨酸羟化酶突变基因类型。结果：分析患者及其父母PAH第7，6，11，3，12，5外显子基因，发现在外显子6中患者及父母的SSCP电泳行为与正常对照均不相同，其中父亲和母亲的电泳条带位置一致，而患儿的电泳条带位置相异。测序结果显示，父亲和母亲的苯丙氨酸羟化酶基因cDNA第526位发生了胞嘧啶被胸腺嘧啶替代的点突变，是R176X突变型杂合子，而患儿的两条染色体都在同一位点发生了突变，为R176X突变型纯合子。结论：在中国回族首次检出苯丙酮尿症R176X突变型纯合予。     

A severe transfusion reaction associated with anti-EnaTS in a patient with an abnormal alpha-like red cell sialoglycoprotein

A 71-year-old woman (Ped) received 3 units of red cells (RBCs), compatible by the indirect antiglobulin test but strongly (4+) incompatible by direct agglutination at 37 degrees C. The next day, her plasma hemoglobin was 1252 mg percent and the direct antiglobulin test (DAT) was weakly positive (IgG and C3). Less than 5 percent of the transfused cells could be detected 48 hours posttransfusion. Her clinical condition deteriorated and renal failure developed. The patient died of pulmonary embolism. Her serum contained a strong (4+) IgM agglutinin and a weakly reactive (microscopically positive) IgG antibody, with anti-EnaTS specificity. EnaFS and EnaTS antigens were severely depressed or absent from the patient's RBCs; the ficin-resistant Ena antigen (EnaFR) appeared to be present. Pretransfusion RBC sialic acid level was 53 +/- 2 percent of normal. The patient's RBC membranes were shown to contain sialoglycoproteins beta and delta by sodium dodecyl sulphate polyacrylamide gel electrophoresis with periodic acid Schiff's base staining, with weak staining of components in the regions corresponding to alpha, alpha 2 and alpha delta. The nature of these components was not identified, but their presence suggested that the patient's RBCs expressed a previously undescribed sialoglycoprotein alpha variant.     

Isoelectric focusing of serum in cystic fibrosis: Failure to distinguish between homozygote and heterozygote sera

Thin-layer isoelectric focusing was performed on samples of sera from patients with Cystic Fibrosis, siblings and obligate heterozygotes (parents), and children without cystic fibrosis (controls). The protein band with an isoelectric point of pH 5.48, previously reported to be absent in homozygote cystic fibrosis sera, was found to have a pI of 5.25. It was present in approximately one-half of the homozygote and heterozygote sera tested, and absent from 18 percent of control sera. The presence of this band is not therefore a reliable marker for the normal gene, and cannot be used to identify the heterozygous carrier for cystic fibrosis.     

Abnormal alkaline phosphatase isoenzymes detected in the serum of elderly patients

Alkaline phosphatase (ALP) isoenzyme analysis of 101,832 serum samples was performed by electrophoresis using cellulose acetate membrane, and abnormal bands at the alpha(1) and alpha(2) globulin positions were detected in 23 samples. The physicochemical properties of these abnormal ALP isoenzyme fractions were examined. In brief, the abnormal fractions were heat-sensitive and inhibited by L-phenylalanine, and neither sialic acid in their polysaccharide chains nor the glycosyl-phosphatidylinositol anchor was detected by the enzyme treatment. The physicochemical properties of abnormal ALP isoenzyme fractions detected in Patients 1 - 22 were similar to those of the adult small intestine type. However, the molecular weight of the adult small intestine type abnormal fractions was smaller than that of the normal fractions. These adult small intestine type abnormal bands at the alpha(1)- to alpha(2)-globulin positions, which were identified in the serum of Patients 1 - 22, were detected in elderly patients. Most of them had various basal diseases such as renal insufficiency, fracture, interstitial pneumonia, and chronic pancreatitis. Some of them had severe diseases such as rectal cancer, descending colon cancer, and septic shock. In Patient 23, the polysaccharide chain had sialic acid and was heat sensitive physicochemical properties that were similar to those of the Kasahara ALP variant.     

常规方法检测血清总胆红素的正确度评价

目的探讨并比较常规方法检测血清总胆红素(T-Bil)的正确度的评价方法。方法参照CLSI EP15-A2文件,用常规方法测定有证参考物质、国际参考实验室室间比对计划(RELA)样本、卫生部临床检验中心室间质评样本T-Bil水平,用BioRad Unity实验室室间质评数据分析本实验室与对等组和方法组间的偏移及标准差指数(SDI),与参考方法比对等方案评估常规测量T-Bil的正确度。结果 NIST SRM 909b参考物质、2010～2011年RELA样本、2012年卫生部临检中心第1次室间质评样本测定结果均在可接受范围内。2012年1～6月Unity实验室间质评数据结果偏移及SDI指数均满足要求。与参考方法比对结果相关性好(Y=1.129 9X-5.734 3,r=0.999 8),小于300μmol/L时,偏移可接受;大于300μmol/L时,偏移较大(10%～13%),不可接受。用去离子水稀释5倍使其浓度约达到50～100μmol/L后重测,偏移可接受,满足临床需要。结论 "单点浓度"的正确度评价具有局限性,临床实验室应分析全测量范围内的正确度情况,与参考方法比对是实现常规测量正确度评价的有效方法之一。     

Elevated cell-free serum DNA detected in patients with myocardial infarction

BACKGROUND: Cell-free DNA is detectable in the circulation. Increased cell-free DNA has been detected in cancer patients and individuals with trauma. We want to know whether patients with myocardial infarction (MI) also have increased cell-free DNA in their blood. METHODS: We used a QIAamp blood kit for DNA extraction from serum and the PicoGreen DNA kit for quantification. DNA patterns of serum DNA were established by gel electrophoresis on 2.5% metarphor gel. RESULTS: The average serum DNA in MI patients (N=55) was 511+/-398 ng/ml, more than 10-fold higher than normal (36.3+/-23.8 ng/ml, n=274). Patients with increased CK-MB (>4%) were associated with highly increased concentrations of cell-free DNA (93.4%). There was no correlation between the concentration of cell-free DNA and the concentrations of CK-MB, troponin I and C-reactive protein. In serial specimens, we found that the cell-free DNA rose early, but peaked behind CK-MB. A slightly diffused DNA ladder could be found with pooled cell-free DNA from MI patients by electrophoresis with the smallest DNA band at only a few hundred base pairs. CONCLUSIONS: Cell-free DNA in MI patients is increased in patients diagnosed with MI, and may complement troponin and CK-MB in a multiple marker test format.     

两种试剂对儿童血清中梅毒反应素检测的比较与结果分析

目的梅毒非特异性血清反应素试验,方法简便快速,可用于梅毒筛查和监测梅毒患者进程和疗效。因此,对本院临检中心使用的检测梅毒患儿血清中反应素两种试剂比较,以选择较好的实验方法,给临床提供更可靠的实验室数据。方法对2009年9月至2010年9月门诊544例患儿血清标本进行两种试剂:梅毒快速血浆反应素诊断试剂(RPR)和梅毒甲笨胺红不加热血清试剂(TRUST)诊断试剂同时检测,其结果进行统计分析。结果两种试剂对544例标本定性试验结果相一致:456例均为阴性,88例均为阳性。88例阳性标本半定量试验:两种试剂85例试验结果相一致,3例标本TRUST试剂抗体滴度比RPR试剂高1个稀释度。结论两种试剂检测的检出结果,差异有统计学意义(P<0.01),具有一致性,对阳性标本滴度的检测TRUST试剂较敏感。     

Transfusion support with RBCs from an Mk homozygote in a case of autoimmune hemolytic anemia following diphtheria-pertussis-tetanus vaccination

BACKGROUND: Autoimmune hemolytic anemia (AIHA) in children, although unusual, is often associated with recent infection. Several reports have identified the diphtheria-pertussis-tetanus (DPT) vaccination as a possible trigger for AIHA. STUDY DESIGN AND METHODS: Life-threatening AIHA was diagnosed in a 6-week-old infant 5 days after receiving a DPT vaccination. The patient required daily transfusion and/or exchange transfusion for 3 weeks. RBCs from an Mk homozygote were found compatible with the patient's autoantibody. Transfusion of RBCs from an Mk homozygote and later RBCs from an individual (K.T.) with a variant glycophorin, Mi.VII, were required to sustain the patient's Hb level until autoantibody production ceased, as evidenced by a fall in antibody titer and the patient's Hct returning to normal. RESULTS: The DAT was positive (3+) with only anti-C3 on presentation. An IgM cold reactive autoantibody with probable anti-Pr specificity and high thermal amplitude (37 degrees C) was identified in the serum. The DAT was no longer positive after transfusion with compatible blood. CONCLUSION: This case represents life-threatening AIHA in an infant, temporally related to a DPT injection and responsive to a combination of immunosuppression and transfusion of rare compatible blood.     

Incidentally detected atrioventricular septal defect in an adult

A 34‐year‐old woman, a liver transplantation donor for her son, was referred to our hospital for preoperative evaluation. She was diagnosed with an incomplete atrioventricular septal defect (AVSD). She underwent double valve repair and patch closure of the defect. Incidental discovery of an asymptomatic AVSD in an adult is rare.     

不孕妇女血清中六项免疫自身抗体检测临床意义的研究

目的 探讨抗精子抗体(ASAb)、抗卵巢抗体(AOAb)、抗子宫内膜抗体(AEAb)、抗透明带抗体(AZAb)、抗滋养层抗体(ATAb)、抗人绒毛促性腺激素抗体(AHCGAb) 等6项指标与女性不孕的关系及临床意义.方法 应用蛋白芯片技术检测131例不孕妇女(实验组)及100名有生育史妇女(对照组)血清中ASAb、AOAb、AEAb、AZAb、ATAb、AHCGAb等6项指标.结果 实验组免疫自身抗体阳性46例(35.11%),对照组1例(1.00%).实验组46例阳性者6项抗体中同时1、2、3项阳性者分别为26、15、5例,阳性率分别为19.85%、11.45%、3.82%;血清中ASAb、AHCGAb、ATAb、AOAb、AZAb、AEAb阳性率分别为21.37% 、9.92%、7.63%、6.11%、5.34、3.82%.结论 所检测的6项免疫自身抗体在不孕妇女中有较高阳性率,6项免疫自身抗体检测对原因不明,以及与免疫因素有关的不孕的诊断有重要临床意义.     

Erythroid accelerating factor detected in serum from rats with drug induced hemolysis.

We have previously observed that an erythroid enhancing activity presents in rat serum in the early stage of drug induced hemolytic anemia. The further studies on biological and physicochemical aspects of this erythroid accelerating factor (EAF) is described in this paper. Hemolytic anemia was induced in rats by single intraperitoneal injection of acetylphenylhydrazine (APH) and serum was obtained from the rats on day 1 after APH injection. It was first fractionated by ultrafiltration on Amicon Diaflo membranes to give a series of fractions lying in the following ranges of molecular weight: 10-30 kDa, 30-50 kDa, 50-100 kDa, and >100 kDa. Among those fractions, largest increase in the number of colony forming unit erythroid CFU-E) colonies was shown in the fraction of >100 kDa that was subsequently fractionated by fast protein liquid chromatography (FPLC) system. EAF activity for CFU-E proliferation was detected in a FPLC fraction corresponding to a molecular weight of about 160 kDa. An addition of EAF significantly increased with dose dependent manner in the number of CFU-E colonies from rat bone marrow mononuclear cells. EAF alone had no burst promoting activity and exhibited no distinct activity to proliferate burst forming unit-erythroid even when interleukin-3 (IL-3) and high concentration (2 U/ml) of erythropoietin (Epo) were added together to the culture. The stimulating effect of EAF on CFU-E was markedly dependent on the presence of adherent cells in the culture. Partially purified protein was relatively heat-unstable (60% at 75 degrees C, 30 minutes) and sensitive to treatment with trypsin and alpha-galactosidase. These results suggest that EAF is a novel factor, possible glycoprotein to reinforce Epo function and is different from various cytokines previously documented because of differences of approximate molecular weight.     

Risk of congenital anomalies detected during antenatal serum screening in women with pregestational diabetes

BACKGROUND: Most studies comparing women with and without pregestational diabetes mellitus have not systematically screened for fetal anomalies in early pregnancy, potentially leading to selection bias. AIM: To evaluate the risk for certain congenital anomalies in women participating in an antenatal maternal screening program. DESIGN: Retrospective cohort study. METHODS: We studied all women who underwent antenatal maternal serum screening in Ontario from 1994 to 2000. Fetal anomalies were documented antenatally by ultrasonography or at autopsy, and postnatally diagnosed birth defects were recorded after 20 weeks gestational age for all live- and stillborn affected infants. We compared the risk of open neural tube defects and urinary tract defects among women with and without pregestational diabetes. RESULTS: Of 413,219 women screened during pregnancy, 2069 (0.5%) had diabetes. Compared to non-diabetic women, the adjusted odds ratios (95%CI) for neural tube and urinary tract defects among women with diabetes were 2.5 (0.9-6.8) and 2.6 (1.4-4.9), respectively. DISCUSSION: Among women who undergo second trimester maternal serum screening, pregestational diabetes is associated with an increased risk of having a fetus with an open neural tube defect or urinary tract disorder.     

荧光定量检测血清HBV—DNA与HBeAg、临床类型的关系

目的 了解乙型肝炎病毒携带、慢性乙型肝炎患病毒含量与血清HBeAg、临床类型的关系。方法 采用荧光定量聚合酶链反应检测乙型肝炎病毒携带和慢性乙型肝炎病人血清HBV－DNA含量。结果 乙型肝炎病毒携带HBeAg阳性组HBV－DNA含量明显高于HBeAg阴性组；乙型肝炎病毒携带HBeAg阴性组中有25％HBV－DNA呈阳性，少数病例HBV－DNA水平还很高；乙型肝炎病毒携带HBV－DNA含量明显视于慢性乙型肝炎。结论 血清HBeAg是反映乙型肝炎病毒复制的良好指标，阳性乙型肝炎携带HBV－DNA含量低于乙型肝炎病毒携带。     

全自动生化分析仪血清胱抑素C参考区间的建立

目的 建立全自动生化分析仪免疫浊度法检测血清胱抑素C(CysC)参考区间.方法 依据美国临床和实验室标准化协会文件的要求进行CysC参考区间验证;免疫浊度法检测480例健康体检者血清CysC浓度,建立本实验室参考区间.结果 20例受检者中有2例受检者检测结果在被验证参考区间外.根据480例健康体检者检测结果,建立参考区间为:50岁以下男性0.81～1.23 mg/L、女性0.70～1.07 mg/L,50岁以上男性0.98～1.89 mg/L、女性0.76～1.61 mg/L.结论 所建立参考区间与试剂盒提供的参考区间有明显差异.血清CysC浓度存在性别及年龄差异,说明建立适合不同本实验室的参考区间十分重要.