发绀患者,男,29岁,因血氧饱和度低(SaO289.3％)而超声心动图和胸片正常就诊.取患者的外周血进行血液学一般检查和溶血相关筛查,高效液相色谱技术(high performance liquid chromatography,HPLC)进行血红蛋白(Hb)分析;采用聚合酶链反应和反向点杂交技术检测17种中国人群常见的β-珠蛋白生成障碍性贫血基因突变和非缺失型α-珠蛋白生成障碍性贫血基因突变;采用跨越断裂点聚合酶链反应(gap-polymerase chain reaction,Gap-PCR)法结合琼脂糖凝胶电泳技术检测缺失型α-珠蛋白生成障碍性贫血基因;采用PCR和DNA测序法对α、β珠蛋白基因(HBA1、HBA2与HBB)进行序列分析.患者的血液学一般检查和其他溶血相关检查均无异常发现;HPLC结果显示存在异常Hb 8.1％(位于4.53 min处,S窗),基因分析显示α2-珠蛋白(hemoglobin A2,HBA2)基因c.175C>T,导致α珠蛋白肽链第58位的组氨酸(His)被酪氨酸(Tyr)替换,为杂合型血红蛋白M病(Hb M病)Boston型.Hb M为结构异常的Hb变异体,该异常使血红素铁稳定在氧化状态(Fe3+),从而影响Hb的正常释氧功能.该病可能会与引起高铁血红蛋白血症的其他原因混淆,如红细胞高铁血红蛋白还原酶系统的遗传性改变等.该例为中国人群中罕见的Hb M病Boston型,临床呈典型终身发绀表现,预后较好,不需要治疗.

Objective To study the hematologic and molecular features of 14 patients with hemoglobin (Hb) variants, so as to provide reference data for its laboratory screening. Methods A total of 1 029 samples were screened by high performance liquid chromatography (HPLC) on the Bio-Rad VariantⅡHPLC system. GAP-PCR and reverse dot blot (RDB) were used to detect common mutation of α and β globin gene in Chinese. DNA sequencing for α and β globin gene was simultaneously performed in samples with abnormal spectrum peak and negative thalassemia gene. Results In 1 029 samples, 10 types of structural Hb variants were detected in14 cases (1.36%), including 1 case of Hb E / β- thalassemia, 1 case of Hb E /α- thalassemia (HbH disease), 2 cases of HbG-Taipei, 2 cases of Hb Q-Thailand, 2 cases of Hb Youngstown, 1 case of Hb Guangzhou-Hangzhou, 1 case of Hb M-Boston, 1 case of Hb G-Siriraj, 1 case of Hb J-Baltimore, 1 case of Hb J-Sicilia and 1 case of Hb Tamano. Conclusion The occurrence of abnormal structural Hb variants with many genotypes in Shanghai is unique. Except for Hb E, Hb Youngstown, and Hb M-Boston, other types of heterozygous are normal in phenotypes, and symptoms such as hemolysis and anemia often occur when other diseases are combined.

Objective: To investigate the application of high performance liquid chromatography (HPLC) in the screening of thalassemia in Shanghai. Methods:The hemoglobin of 135 patients with suspected thalassemia were analyzed by using HPLC. Gene detection of thalassemia by GAP-PCR and reverse dot blot (RDB) were simultaneously performed. Results: Using gene detection as a gold standard, when HbA2>3.7%was taken as the cutoff value for diagnosis of β-tha-lassemia carriers by HPLC, the sensitivity and specificity were 100%, and 98.41%, and the positive predictive value and negative predictive value were 98.0%and 100%, respectively. If<1.7%was taken as the cutoff value of HbA 2 for diagno-sis ofα-thalassemia (HbH disease) by HPLC, the indice above-mentioned were 100%, 100%, 100%, 100%, respectively. Out of 135 samples, there were 8 cases being detected a HbH peak. Conclusion: Results of HPLC detection are in ex-cellent agreement with those of the gene detection in the diagnosis of β-thalassemia and HbH disease, while certain false negative rate may occur in the diagnosis of minor α-thalassemia orα-thalassemia trait.

Objective To assess the value of reticulocyte hemoglobin equivalent (Ret-He) in the differential diagnosis of microcytic hypochromic anemias,including iron deficiency anemia (IDA),anemia of chronic disorders (ACD),iron deficient combined with ACD and thalassemia.Methods Ret-He was measured by Sysmex XT-4000i in 43 IDA patients,31 ACD patients,66 iron deficient combined with ACD patients,19 thalassemia patients,and 50 normal controls.Results The mean value of Ret-He in IDA was below normal value and was also significantly lower than those of the other four groups (P0.05).Ret-He levels in all the microcytic hypochromic anemia groups were significantly lower than that of normal controls(P0.05).No significant difference was found in Ret-He levels among ACD group,iron deficient combined with ACD group and thalassemia group (P0.05).When 20.9 pg was defined as the Ret-He cut off value for diagnosing IDA,the sensitivity was 57.1% and specificity was 83.7%.The area under receiver operating characteristic curve (AUC)of Ret-He and SF was 0.759 and 0.641,respectively.Conclusions Ret-He is an effective assistant parameter for sTfR in diagnosing IDA.It can also be looked as an important index in the differential diagnosis of IDA from ACD,iron deficiency combined with ACD and thalassemia.

Objective To study the significance of atypical lymphocyte detection by the high fluorescence lymphocyte(HFL) area(HFL-count) from the XT-4000i hematology analyzer leucocyte differential(DIFF) channel,and compare the results of manual microscopy and epstein barr virus-viral capsid antigen(EBV-VCA) detection.Methods HFL-count was analyzed by DIFF channel in total 230 samples,and the results were compared with those of manual microscopy.Serum EBV-VCA detection based on the enzyme immunoassay.Results The false positive rate of atypical lymphocytes by XT-4000i hematology analyzer was 13.6%,and the false negative rate was 2%.Conclusions There is a certain validity for atypical lymphocyte detection by XT-4000i,and XT-4000i can improve the efficiency of clinical diagnosis.

Objective To analyse the types of glucose-6-phosphate dehydrogenase(G6PD) gene mutations in patients with G6PD deficiency.MethodsThe blood samples of 49 patients with G6PD deficiency and 100 normal subjects confirmed by G6PD activity detection were collected.Sequence analysis was performed on 12 exons from 2 to 13 by PCR and DNA sequencing.ResultsTwelve different missense G6PD mutations were detected,and the three most common mutations were G6PD G1388A(26.5%),G1376T(28.6%) and A95G(14.3%).The G6PD activity of these individuals was 5% to 18% of that of normal subjects.The clinical manifestations of these subjects were acute hemolytic anemia trigged by ingestion of fava bean and neonatal jaundice.The other mutations included C1024T(4.1%),C1225T(2.0%),C1159T(2.0%),G487A(4.1%),G392T(4.1%),G1160A(6.1%),G871A / C1311T(4.1%) and C 406T/C1311T(2.0%).A novel missense mutation(G691 C) in exon 7 of the G6PD gene predicting a single amino acid substitution,Ala231Pro was identified.The same gene mutations were not detected in samples of normal subjects.ConclusionG6PD gene G1388A,G1376T and A95G are the three most common types of G6PD gene mutations in patients with G6PD deficiency.A novel missense mutation G691C is found,which produces an Ala231Pro substitution and leads to significantly decreased G6PD activity in red blood cells.

Objective To detect the argyrophilic proteins in nucleolar organizer regions(Ag-NORs) that express rDNA and rRNA proliferation of T lymphocytes before chemotherapy and after complete remission(CR) in children with primary acute leukemia(AL).Methods The argyrophilic granules area of NOR/nuclear area(I.S%) of T lymphocytes was detected by image analysis system in peripheral blood of 42 patients before chemotherapy and after CR and 30 normal children.Results I.S% in the patients before chemotherapy(5.06%±1.36%) were significantly lower than those in the healthy donors(7.51%±1.06%)(t=8.238 P0.001).I.S% in the patients after CR(7.17%±0.98%) were significantly higher than those before chemotherapy(t=7.073 P0.001).However,no significant difference was encountered in I.S% between the patients after CR and the normal children(t=1.403 P0.05).Conclusion These results suggest that decrease of Ag-NORs expresses the evidence for tumour induced suppression of immune function of T cells in children with AL prior to treatment.

目的通过对急性白血病( AL)患儿 T淋巴细胞( T细胞)表示 rDNA和 rRNA活性的嗜银蛋白的测定,了解患儿 T细胞的功能状态,并分析其与 T细胞亚群之间的关系.方法应用图象分析法测定 31例初发未化疗 AL患儿及 30例正常儿童(对照组)外周血 T细胞核仁组成区嗜银蛋白(银染区) /细胞核面积( I.S%),用流式细胞仪测定 T细胞亚群.结果 31例 AL患儿 I.S%值( 5.78%± 2.66%)较对照组( 7.51%± 1.06%)下降( P<0.01).T细胞亚群测定中, CD4+ ( 27.50%± 12.56%)较正常值( 35%± 2.55%)下降( P<0.05); CD8+ ( 26.11%± 12.35%)与正常值( 25%± 2.04%)比较差异无显著性( P>0.05); CD4+ /CD8+ ( 1.149± 0.51)较正常值( 2± 0.26)下降( P<0.001).结论在初发未化疗的 AL患儿中, T细胞嗜银蛋白含量下降,代表患儿 T细胞免疫功能的 CD4+和 CD4+ /CD8+值降低,显示患儿 T细胞活性的改变与 T细胞亚群的变化相符,患儿存在 T细胞免疫功能降低.

目的:评估血红蛋白分析对地中海贫血(Thal)诊断能力.方法:用HLC-723G7自动血红蛋白分析仪对以临床表现流行病学资料和醋纤薄膜电泳等确定的115例Thal患者进行血红蛋白分析,结合分子诊断评估诊断价值.结果:以HbA2≥5.5%作为血红蛋白分析对β-Thal的诊断标准,则与DNA分析的符合率为81.5%;以HbA2≤2.5%为仪器对α-Thal的诊断标准,则与DNA分析的符合率为92.5%.结论:血红蛋白分析可提供α、βThal的诊断信息,若能结合更多的临床和DNA分析数据,可作为α、β地中海贫血的确诊手段.

Objective To investigate the cell immunity in eosinophilia patients. Methods 100 patients' venous blood in which eosinophil was over 5% and 145 patients' venous blood in which monocyte was over 8% were analyzed the Ag-NORs expression of the lymphocytes by a KL imaging system. Results The Ag-NORs expression of the T lymphocytes in eosinophilia was higher than that in controls. Conclusions The cell immunity in eosinophilia is enhanced, which can be screened throngh automated hematology analyser.

Purpose:To study the manifestation and clinical significance of rDNA transcription activity analysis of T lymphocytes in cancer patients.Methods:We analyzed the activity of the Ag NORs as one of the rDNA transcription activities in T cell of 100 normal human and 263 tumor patients by KL image system.Results:There were significant differences in the IS%of T lymphocytes in peripheral blood between normal human and cancer patients( P 0 01). The same results can be observed between the patients with tumor and without tumor residual as well as in different ages of normal human ( P 0 01).There were no significant differences in normal persons or cancer patients of different ages as well as in different position and the cell type of cancer ( P 0.05).Conclusions:There were significant differences in IS% of T lymphocytes in peripheral blood between normal human and cancer patients. But there was no relationship with the gender of the cancer patients, position and the cell type of tumor. So it can be one of the approaches to diagnose cancer and to monitor the progress. [

目的 探讨血红蛋白Lepore病 （HbLepore）的诊断、治疗及发病机理。方法 用SDS聚丙烯酰胺凝胶电泳（PH8.7、PH6 .2 ）及Hb肽链分析法检测此病 ,用皮质激素等治疗 ,同时给予切脾。结果 病人获得缓解。结论 HbLepore是一种在亚太地区罕见、临床表现较重的特殊珠蛋白生成障碍性血红蛋白病。

Objective: To estimate the clinical application of antigen and activity of t-PA and PAI-1 . Methods: We studied 189 subjects of NIDDM and grouping(insulin resistance, coronary heart disease, insulin resistance with coronary heart disease, pure NIDDM), the concentrations and activities of t-PA and PAI-1 were measured using commercial kits. Results: There are differences between each NIDDM groups and normal group of activities of PAI-1 and t-PA, and also of the ratio of PAI-1 . Conclusion: The activities of PAI-1 and t-PA are more sensitivity in these index, and the ratios of t-PA/PAI-1 have more valve in clinical application

Objective: To investigate the status of platelet activation and the injury of vascular endothelium in N1DDM patients. Methods: The plasma level of 11-DH-TXB2αand 11-β-PGF2α were detected and analysed in 21 health controls and 50 patients by the method of competitive EIA. Results: The plasma level of 11-DH-TXB2 patients with NIDDM was significantly increased compared with the normal controls. It was divided into groups according to clinical manifestations and there were various plasma levels of 11-DH-TXB2 in different groups. Especially, the patients with insulin resistance were found to have higher level of plasma 11-DH-TXB2. Plasma levels of 11-β-PGF2α were not significant in each groups including the controls. Conclusions: The platelet was highly activated in the patients with NIDDM. High plasma level of 11-DH-TXB2 and massive platelets activation were closely related to NIDDM patients with insulin resistance and injury of vascular endothelium. Plasma level of 11-DH-TXB2 was detected as a reliable indicator of evaluating the status of platelet activation in patients with DM.

抗磷脂抗体(APA)是一类异质免疫球蛋白,在体内可以与带负电的磷脂结合,在参与凝血过程时, 影响多种依赖磷脂的反应体系,可引起凝血功能的紊乱.目前认为抗磷脂抗体与血栓形成关系甚为密切[1,2].抗磷脂抗体可分为狼疮样抗凝物(lupus-type anticoagulants, LAC)和抗心磷脂抗体(anticardiolipin antibodies, ACA)两大类.近年来对恶性血液病的研究发现,在恶性血液病的病程进展中常可以伴有血栓和(或)弥散性血管内凝血(DIC)的出现.

Objective To establish a standard principle for using various ELISA kits, the reliability on detection of platelet reactive antibodies was investigated. Methods Type IgG platelet reactive antibodies were determined in 30 ITP patients, 10 patients with active chronic hepatitis B, 5 patients with auto immune conditions, 6 infectious patients and 20 controls by different ELISA kits or MAIPA. Results There are significant difference in positive rates of platelet reactive antibodies tested with different kits. Influence effects involved different platelet lysate way, different temperature of platelet preoperated and with or without(NH 4) 2C 2O 4. MAIPA is more sensitive and specific than ELISA in detection of platelet reactive antibodies. Conclusion The principle of detection of platelet reactive antibodies by ELISA requires to preoperate platelet in 4℃ and to do platelet lysate by Nonidet P 40. Also it is necessary to avoid (NH 4) 2C 2O 4.To replace ELISA with MAIPA is more reliable.

在原发性高血压、老年病和其他高凝状态研究中发现,脑腔隙性梗死患者存在高凝状态,并发现部分病例有明显的FⅫ活性的减低,因而将FⅫ活性与血栓形成、血管变性、不确定性的脑腔梗一起研究,以了解脑腔梗患者纤溶系统改变及其在血栓形成中的作用.

白介素6(IL-6)是一种多功能的细胞因子,除涉及感染、炎症等急性期反应外,对T、B细胞的发育、骨髓造血细胞增殖均有很重要的作用[1].自多发性骨髓瘤患者血浆IL-6水平增高被发现后,又发现IL-6血浆水平增高还见于弥散性血管内凝血(DIC),并且与DIC的严重程度、治疗反应和DIC是否伴有脏器损伤有关[2].

白介素6(IL-6)是一种多功能的细胞因子，除涉及感染、炎症等急性期反应外，对T、B细胞的发育、骨髓造血细胞增殖均有很重要的作用[1]。自多发性骨髓瘤患者血浆IL-6水平增高被发现后，又发现IL-6血浆水平增高还见于弥散性血管内凝血(DIC)，并且与DIC的严重程度、治疗反应和DIC是否伴有脏器损伤有关。IL-8属于α型趋化因子，在其多种生物特性中，有一种趋化中性粒细胞的功能。在整合素参与下，作用于血小板和内皮细胞，使CD16和CD18的表达和IL-6一样卷入凝血和血栓形成的过程。近又证实单核细胞表面可表达白介素6受体(IL-6R)，单核细胞也可能受到IL-8的趋化作用。单核细胞是血液细胞中最重要的组织因子(TF)来源的细胞，TF在DIC和其他血栓形成病变中的作用是不容置疑的[2]。本文就IL-6和IL-8对外周血单核细胞体外培养中TF表达的影响，研究IL-6、IL-8与单核细胞、TF在血栓形成中的相互关系。1 材料与方法1.1 材料1.1.1 外周血单核细胞(PBMC) 取自10名正常健康志愿者外周血。1.1.2 试剂 rhIL-6、hIL-6 MoAb和rhIL-8、IL-8 Mo-Ab购自STAGO公司。TF ELISA检测试剂盒购自Immunotech公司。Ficoll液、Hypague液购于Sigma公司。RPMI1640培养液、NCS购于Sigma公司。1.2 方法1.2.1 外周血单核细胞分离 采用密度梯度离心法。1.2.2 外周血单核细胞原代培养 将2×105ml-1的外周血单核细胞悬浮于20％的NCS RPMI1640培养液中，以5％CO2 37℃培养24h。将PBMC培养液用水平离心机以3 000 r/min，离心10 min，取上清液待测。1.2.3 上清液TF抗原含量检测 以ELISA方法测定外周血单核细胞培养(加或不加rhII-6、hIL-6 MoAb和rhIL-8、IL-8 MoAb)上清液中的TF抗原含量。