免疫性输血反应的调查及预防研究

目的　对各类输血反应进行调查研究 ,同时建立输血前免疫血液学检查新技术 ,以预防和治疗免疫性输血反应。方法　对上海地区 13家医院的 30 776名输血病人进行调查 ,并应用改良Polybrene、简易致敏红细胞血小板血清学试验 (SEPSA)等新技术 ,进行同种特异性抗体的筛选、鉴定及交叉配型试验。结果　发生各类输血反应5 0 8例 (1.6 5 % ) ,其中检出引起免疫性输血反应的同种抗体 6 4例 (12 .6 % ) ,抗体特异性分别为血小板抗体 (包括HLA) 5 1例 ,红细胞不规则抗体 13例 ;各项新技术的应用对预防免疫性输血反应的发生具有显著相关性 (χ2 >3.84 ,P <0 .0 5 )。结论　使用研究建立的输血前免疫血液学新技术能有效地诊断、预防和治疗免疫性输血反应的发生 ,提高了输血的安全性和有效性     

上海地区人群中部分稀有血型筛选

目的了解上海地区人群中部分稀有血型的频率和分布情况,用于解决临床上稀有血型的用血问题。方法利用稀有单克隆、多克隆抗血清,对Kell血型系统的K0,Miltenberger血型系统的Mur＋,Duffy血型系统的Fya-,高频率抗原中的Lan-、Vel-进行筛选;用2 mol/L尿素进行Kidd血型系统的Jk（a-b-）表型筛选。筛选方法包括试管法间接抗人球蛋白试验（IAT）,U型96孔微量板IAT和U型96孔微量板盐水直接离心法试验。结果从24 093名献血者中发现1例K0表型;在2 970名献血者中检出15例Mur＋;从200名献血中,检出1例Fya-;从6 153名献血者中发现2例Vel-;在300名献血者中发现了1例Lan-;在102 760名献血者中,成功检出4例Jk（a-b-）。结论Mur＋、Lan-、Jk（a-b-）表型频率明显高于白种人;Fya-表型频率低于白种人。     

环磷酰胺抑制作用下小鼠对人红细胞抗原的选择性耐受

目的　以环磷酰胺 (CY)作为免疫抑制剂 ,诱导小鼠产生对人红细胞血型抗原的选择性免疫耐受。　方法　1 0 2只 Balb/c小鼠分成 1 7组 ,在 2× 1 0 7人红细胞免疫注射后 ,分别以不同 CY剂量和给药时间给予抑制。并通过直接和间接血凝反应测定小鼠免疫应答所产生的抗人红细胞抗原的 Ig M和 Ig G型抗体效价。　结果　在 5 0～ 1 5 0 mg/kg CY剂量范围内 ,免疫注射当天至免疫后第 4天 (d0～d4)均可对小鼠的免疫应答产生抑制。而诱导小鼠产生对人红细胞抗原完全耐受的最佳 CY抑制时间和剂量分别为免疫注射后的第 1天和 1 0 0 mg/kg。　结论　 CY能有效抑制小鼠对人红细胞免疫所产生的应答 ,选择适当的给药时间和 CY剂量对耐受的建立十分重要。本项研究对诱导小鼠免疫系统产生对人红细胞血型弱抗原的应答具有探索意义。     

小鼠调节性树突状细胞分泌的外泌体诱导免疫耐受的实验研究

本研究探讨小鼠调节性树突状细胞（rDC）分泌的外泌体（regulatory exosomes，rDex）诱导免疫耐受的作用，并与正常未成熟树突状细胞的外泌体（immature exosomes，iDex）诱导免疫耐受的作用进行比较。取C57BL／6（H-2^b）小鼠骨髓细胞诱导未成熟树突状细胞（iDC），TGF—β1联合IL-10诱导调节性树突状细胞（rDC），流式细胞术检测两种DC的表型。采用超速离心结合膜超滤的方法分别提取rDex和iDex。建立小鼠皮肤移植模型．以C57BL／6小鼠为供者，以BALB／c（H-2^d）小鼠为受者，按对受者处理的不同实验分3组，iDex组术前7、3天受者经尾静脉注射10μg供者的iDex，rDex组术前7、3天受者经尾静脉注射10μg供者的rDex，另设PBS对照组，观察移植皮肤存活情况。通过单向混合淋巴细胞反应（MLR）观察供者及非亲缘异基因供者DBA／2对同种异基因小鼠T细胞增殖情况。结果表明：TGF—β1、IL-10可下调DC表面共刺激分子CD80、CD86、CD40的表达。移植皮片平均存活时间（MST），在对照组为7、8天，iDex组为10．7天，rDex组为18．8天；iDex组的移植皮片MST明显长于对照组（P〈0．05）；rDex组的移植皮片MST明显长于iDex组（P〈0．01）。MLR结果证明iDex组和rDex组B／C小鼠均对C57小鼠的脾细胞产生特异性耐受，尤其是rDex组；对非亲缘异基因DBA供者的脾细胞两组仍表现出强烈的免疫应答。结论：iDex和rDex均有诱导免疫耐受的作用，且rDex作用较iDex作用好。     

220例患者血型不规则抗体分析

目的 对上海地区患中不规则抗体检出情况进行回顾性总结。方法 对2000年以来上海各医院送捡的免疫血液学标本2149例，应用盐水、抗球蛋白法、聚凝胺法检测不规则抗体。结果 共鉴定确认血型不规则抗体220例。其中自身抗体66例、确定同种特异性抗体154例，分别为：抗-D 41例、抗-E 37例、抗-cE 19例、抗-Ce 4例、抗-CD 3例、抗-c 4例、抗-M 15例、抗-N 3例、抗-PI 2例、抗-Le^a 1例、抗-Le^b 4例、抗-Le^x 1例、抗-Mur 3例、抗-S 2例、抗-HI 11例、抗-Jk^b 1例、抗-Fy^b 1例、抗—Tj^a 1例、抗-I 1例。结论 检出的不规则抗体与国外数据存在较大差异，国外常见的抗—K、抗—Fy^a在国内较少见。产生抗体的患(不包括孕妇)以女性居多。     

The prevalence of factor VIII inhibitors and genetic aspects of inhibitor development in Chinese patients with haemophilia A

Summary. The prevalence of inhibitors in Chinese haemophiliacs has not yet been reported. The aim of this study was to identify the prevalence of factor VIII (FVIII) inhibitors among haemophiliacs who are treated only with plasma‐derived FVIII (pdFVIII), cryoprecipitate or fresh frozen plasma (FFP), and tried to explore the relationship between the generation of inhibitors and particular FVIII deficiency genotypes. Clinical information and blood samples of 1435 patients with haemophilia A (HA) were collected by six haemophilia centres in China. The Nijmegen modification of the Bethesda assay was used to detect inhibitors. Multiplex PCR, long‐range PCR and direct sequencing were performed for genotyping. The overall prevalence of inhibitors in Chinese HA patients was 3.9% and the prevalence of severe haemophiliacs was 4.3%; 18 of the 56 patients with inhibitors had high titres. A total of 38 different mutations were identified in the 55 patients with inhibitors, including 15 intron 22 and 3 intron 1 inversions, seven large deletions, 14 small deletion/insertions, seven nonsense mutations, one splice site mutations and eight missense mutations. Of 38 mutations, 28 were novel. Patients with large deletions and nonsense mutations were prone to have high titre inhibitors, with incidence rates of 57.1% (4/7) and 42.9% (3/7), respectively. In conclusion, the prevalence of inhibitors in Chinese HA patients is much lower than that reported for other ethnic groups and the large deletion and nonsense mutations are high risk factors for high titre inhibitor development.     

不同检测方法在无偿献血筛选ALT中的应用评价

[目的]寻求一种适合应用于血液中心无偿献血血清丙氨酸氨基转移酶(ALT)筛选的检测方法,以达到节约血源,大幅度降低因ALT不合格造成的血液报废。[方法]用MissionTMC100干式生化分析仪(艾康)、Reflotron plus快速全自动干式生化分析仪(罗氏)、microlab 300半自动生化分析仪(威图)3种仪器分别测100例献血者的ALT,并分别与日立7600-010全自动生化分析仪进行比较。[结果]与全自动生化仪测试结果相比,艾康干式法χ2=2.25,相关性r=0.9550;罗氏χ2=4.167,相关性r=0.9724;威图半自动χ2=0.25,相关性r=0.9695。[结论]威图半自动生化分析仪结果良好,相关性好,与日立7600-010相比无明显差异;艾康与罗氏应适当调整临界值,艾康应调为35 U/L,罗氏应调为34 U/L。     

Two novel HLA-A alleles: HLA-A*31:01:09 and HLA-A*33:30

The HLA-A*31:01:09 differs from the closest allele HLAA*31:01:02 by a C to T silent substitution at position 447 in exon 3. The HLA-A*33:30 differs from the closest allele HLA-A*33:03:01 by a single nucleotide substitution at position 453 in exon 3, leading to a change of Arg 131 Cys.     

Effects of genistein on secretion of extracellular matrix components and transforming growth factor beta in high-glucose-cultured rat mesangial cells

The ideal treatment for diabetic nephropathy should slow the progress of renal failure, delay the initiation of dialysis, and protect residual renal function in patients receiving dialysis. Renal mesangial cells play an important role in these processes. In the current study, we investigated the effects of genistein on rodent renal mesangial cells cultured in a high-glucose environment. Since overexpression of the extracellular matrix (ECM) components (type IV collagen and fibronectin) and transforming growth factor beta (TGF-β) have been previously implicated in the development of the renal glomerulus damage of diabetic nephropathy, we included these substances in our study. The results showed that high concentration of glucose (450 mg·dl⁻¹) stimulated the synthesis of type IV collagen and fibronectin and the secretion of TGF-β. These responses were attenuated by genistein (≥5 μmol·l⁻¹) in a dose- and time-dependent manner. In conclusion, we demonstrated that genistein could inhibit the secretion of ECM components and the expression of TGF-β at both the protein and mRNA levels. These findings should be followed up by further studies and clinical trials to verify the potential therapeutic effects of genistein on diabetic nephropathy.