An anti-EnaTS detected in the serum of an MiI homozygote

The serum of EH reacted with all red cells (RBCs) except her own, ficin- or trypsin-treated red cells, and En(a-) red cells. This reactivity defined an anti-EnaTS specificity. The red cells of the proposita typed as M-N+S-S+, Vw+Mur-Hil-Hut-Anek-Lane-, Wr(a-b+), EnaKT+. Red cells of five relatives were Vw+ and positive with her serum. Titration studies suggest that EH is genetically an MiI homozygote and that her Vw+ relatives are MiI heterozygotes. There is no history of consanguinity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting studies have agreed with the serologic observations. A variant sialoglycoprotein of faster mobility than normal glycoprotein A, but no normal glycoprotein A, was detected on her red cells. Treatment with N-glycanase did not alter the mobility, which indicated that there was no N-glycosylation of residue 26. These findings are in agreement with the reported properties of the Mi.I-specific glycoprotein A. The relatives' Vw+ red cells showed the variant sialoglycoprotein and normal glycoprotein A. EH appears to be the first reported MiI homozygote.     

Time-dependent loss of surface complement regulatory activity during storage of donor blood

The survival of transfused red cells (RBCs) diminishes with time of in vitro storage in blood banks, but the molecular mechanisms underlying the slow but incessant deterioration are incompletely understood. To investigate the possibility that impaired resistance to autologous complement attack could play a role in this phenomenon, packed RBCs stored for variable periods were assayed for decay-accelerating factor (DAF) and CD59, two glycoinositol-phospholipid (GPI)-anchored, membrane- associated complement regulatory proteins that function physiologically to protect blood cells from autologous complement activation on their surfaces. Immunoradiometric and flow cytometric assays employing DAF and CD59 monoclonal antibodies showed that levels of both surface proteins gradually declined over 6 weeks. Digestion analyses with phosphatidylinositol-specific phospholipase C, an enzyme that releases GPI-anchored proteins from cell surfaces, showed that DAF and CD59 molecules with GPI anchors containing unacylated inositol were preferentially lost. These findings suggest: 1) that DAF and CD59 molecules with acylated GPI anchors are more stable in RBC membranes than are molecules with unacylated GPI anchors, and 2) that DAF and CD59 loss may participate with other membrane alterations that occur during in vitro storage in compromising the survival of transfused cells.     

Amplatzer封堵器的生物学性能及经导管介入治疗房间隔缺损

目的:评价经导管置入Amplatzer封堵器治疗继发孔房间隔缺损的治疗效果。方法:①选择2002-08/2006-04在兰州市第一人民医院心外科住院的继发孔型房间隔缺损65例,男26例,女39例;平均年龄(18±8)岁;平均房间隔缺损直径(19.3±7.2)mm。纳入患者对手术方案知情同意。②手术所用封堵器为美国公司的Amplatzer房间隔缺损封堵器,是一种新型的适于关闭二孔型房间隔缺损的装置,它由具有自膨胀性的双盘及连接双盘的腰部三部分组成。双盘状结构恢复记忆形状后可以稳定封堵房间隔缺损的边缘部分,降低残余分流的发生率。③根据选择封堵器大小的方式(即球囊测量或经胸超声心动图直接观察)将患者分为球囊测量组38例和经胸超声心动图测量组27例。均在透视及经胸超声心动图监视下经导管置入Amplatzer封堵器封堵房间隔缺损。同时测量患者缺损扩张直径、封堵器大小,记录X射线透视时间和手术时间。④术后即刻、24h、3个月及1年分别行经胸超声心动图、心电图及X射线检查评价治疗效果。⑤超声心动图显示完全无分流为无分流;残余分流血流宽度≤1mm为微量分流;血流宽度1.0～2.0mm为少量残余分流;血流宽度2～4mm为中量残余分流;血流宽度>4mm为大量残余分流。⑥组间计量资料差异比较采用两个独立样本t检验,组间手术效果比较采用两个独立样本的等级资料秩和检验。结果:①技术成功率:65例房间隔缺损患者,64例封堵器置入成功,技术成功率为98%。②选择封堵器直径:球囊测量组缺损扩张直径为(20.4±6.1)mm,选择的封堵器直径为(21.6±5.7)mm,与经胸超声心动图测量组相近[(22.5±4.3),(25.1±4.9)mm,P>0.05]。③术后残余分流情况:术后即刻经胸超声心动图显示,球囊测量组35例完全无分流,经胸超声心动图组有23例,差异不明显(P>0.05);术后24h,球囊测量组36例完全无分流,经胸超声心动图组有24例,差异不明显(P>0.05);术后3个月,球囊测量组37例完全无分流,经胸超声心动图组有25例,差异不明显(P>0.05);术后1年完成随访的52例患者均未见封堵器移位及房间隔缺损再通。④X射线平片检查:全部显示肺血减少,右心房、室缩小。结论:封堵器直径比球囊测量的房间隔缺损扩张直径大1.0～2.0mm,比超声心动图测量的大2～6mm封堵效果好,成功率高。     

Biopterin synthesis defect. Treatment with L-dopa and 5-hydroxytryptophan compared with therapy with a tetrahydropterin.

We have identified a generalized deficiency of monoamine neurotransmitters in a patient with a defect in biopterin synthesis. Neurotransmitter precursors (L-3,4-dihydroxyphenylalanine [L-dopa]; 5-hydroxytryptophan [5-HTP] and a tetrahydropterin [6-methyltetrahydropterin (6MPH4)] were investigated for their ability to normalize monoamine neurotransmitter metabolism. Before treatment, the concentrations of dopamine (DA), norepinephrine, epinephrine, and six monoamine metabolites were very low or undetectable in plasma, cerebrospinal fluid, or urine. L-Dopa and 5-HTP replacement was begun at age 7 mo. This therapy generally corrected the deficiency of monoamines and their metabolites, and improved neurological development until the age of 25 mo. Despite these benefits, the intermittent administration of L-dopa could not produce a stable improvement of acute neurological function or DA metabolism. In the 3 h after L-dopa administration, plasma DA and the motor activity and alertness of the patient rose and fell in parallel. Doses of L-dopa that were clinically optimal produced normal plasma levels of norepinephrine and epinephrine, but excessive concentrations of DA and its metabolites. Furthermore, the clinical and biochemical effects of L-dopa were inhibited by phenylalanine and 5-HTP, respectively, demonstrating that these amino acids have antagonistic pharmacological effects. Physiological correction of the monoamine deficit and the hyperphenylalaninemia of this disorder was attempted at age 35 mo using high doses (8-38 mg/kg per d) of 6MPH4. 6MPH4, a synthetic analogue of tetrahydrobiopterin, controlled the hyperphenylalaninemia. Significant concentrations of 6MPH4 were obtained in the cerebrospinal fluid; no neurological improvement or stimulation of monoamine synthesis in the central nervous system was detected. These findings indicate the complexity in replacement therapy with L-dopa and 5-HTP, but suggest that this treatment may be partially effective in biopterin-deficient patients who are unresponsive to high doses of tetrahydropterins.     

转脑啡肽基因移植细胞微囊化对大鼠慢性周围神经损伤的镇痛作用

目的：观察微囊化转大鼠脑啡肽原基因（pENK）细胞移植对慢性脊神经压榨性损伤大鼠的镇痛效应. 方法：实验于2006-03/2007-04在郑州大学医学重点实验室完成.①实验方法：通过反转录-聚合酶链反应技术可获得大鼠pENK基因,Hind Ⅲ,ClaⅠ双酶切后,同相应双酶切的pLNCX2载体大片段连接,构建成pENK基因反转录病毒载体pLNCX2-Enk,然后用脂质体法将该载体转染PT67细胞,G418筛选,获得携带pENK基因高滴度反转录病毒产毒细胞系.用海藻酸钠-多聚赖氨酸-海藻酸钠微囊包埋后,进行体外培养,定期检测微囊化细胞活性和pENK分泌量变化.同时,将转基因细胞移植于慢性脊神经压榨性损伤大鼠的蛛网膜下腔.②实验分组：SD大鼠60只,其中53只按照Bennett和Xie法制作大鼠慢性左侧坐骨神经压迫性损伤模型,其中42只造模成功.术后1周,将动物按随机数字表法分为3组,每组16只：微囊化转基因细胞移植组、空囊移植组和阴性对照组.微囊化转基因细胞移植组、空囊移植组分别植入微囊化细胞悬液80 μL（约300个微囊）、空微囊悬液80 μL（约300个空囊）,阴性对照组不注射任何悬液.③实验评估：术后2周和8周行甲醛实验,在大鼠一侧后爪掌侧皮下注射体积分数为0.05的甲醛50 μL,观察其注射后1 h内的痛反应.采用Abbott等所推荐的以缩腿及舔爪时间之和作为行为学反应的指标.注射甲醛后,立即记录大鼠1 h内每5 min的缩腿及舔爪时间. 结果：①术后2周甲醛实验：空囊移植组第一时相的急性痛阶段（注射后5 min）和第二时相的慢性痛阶段（注射后41～45 min）大鼠的缩腿及舔爪时间都少于微囊化转基因细胞移植组（P＜0.01）.而在静息期,3组大鼠的缩腿及舔爪时间差异无显著性意义（P＞0.05）.②术后8周甲醛实验：3组大鼠的痛觉行为都呈明显的双时相反应.除了静息期（注射后5～15 min）,微囊化转基因细胞移植组在各时间点上大鼠的缩腿及舔爪时间均低于空囊移植组（P＜0.05）.微囊化转基因细胞移植组大鼠的缩腿及舔爪时间在第一时相的急性痛阶段和第二时相的慢性痛阶段都低于空囊移植组（P＜0.05）. 结论：微囊化转pENK基因细胞移植对慢性神经痛大鼠有一定的镇痛作用,有望成为运动员慢性疼痛治疗的新方法.     

Demographic characteristics and prevalence of serologic markers among donors who use the confidential unit exclusion process: the Retrovirus Epidemiology Donor Study

BACKGROUND: Most blood centers utilize a confidential unit exclusion (CUE) process, intended to reduce the risk of transfusion-associated infectious diseases by allowing high-risk donors confidentially to exclude their blood from use for transfusion. The effectiveness of this method remains controversial. STUDY DESIGN AND METHODS: Confirmatory or supplemental test results for antibodies to human immunodeficiency virus, human T-lymphotropic virus type I, and hepatitis C virus, as well as hepatitis B surface antigen and syphilis and screening test results for antibodies to hepatitis B core (antigen) and alanine aminotransferase levels were obtained for approximately 1.8 million units donated during 1991 and 1992 at five blood centers within the United States. The prevalences of these infectious disease markers in units that the donors confidentially excluded (CUE+) and units that the donors did not exclude (CUE-) were calculated and examined within demographic subgroups. RESULTS: Units that were CUE+ were 8 to 41 times more likely to be seropositive for antibodies to human immunodeficiency virus and hepatitis C virus, hepatitis B surface antigen, and syphilis and three to four times more likely to react for antibody to hepatitis B core (antigen) or to have elevated alanine aminotransferase levels than units that were CUE- (p < 0.001). The positive predictive value of CUE (the percentage of CUE+ units that were confirmed seropositive for any marker) was 3.5 percent, and the sensitivity of CUE (the percentage of confirmed-seropositive units that were CUE+) was 2.3 percent. CONCLUSION: The current CUE process has low sensitivity and apparently low positive predictive value, and in many cases, it appeared that donors misunderstood it. Yet, CUE was not a “random process,” as CUE+ units were more likely to be seropositive for any infectious disease marker than CUE- units. This suggests that efforts to improve the CUE system may be warranted. As risk factors for transfusion-transmitted infection become more difficult to identify by history-based screening, however, such efforts may have limited effect.     

The molecular basis of renal amyloidosis in Irish-American and Polish- Canadian kindreds

Hereditary amyloidosis of an unusual form has been reported in two separate kindreds; one was Polish-Canadian and the other was Irish- American (Am J Med 1975; 59:121 and Trans Assoc Am Physicians 1981; 94:211). In both kindreds, affected members developed hypertension and nephrotic syndrome due to amyloidosis in their forties or fifties, but the genetic background responsible for the condition has been left undetermined. To identify the genetic defect in these kindreds, a portion of exon 5 of the fibrinogen alpha-chain gene in members of these kindreds was examined for a mutation by single-strand conformation polymorphism analysis and direct DNA sequencing. DNA analyses revealed an A-->T transversion at the second base of codon 526 of the fibrinogen alpha-chain gene in both of these kindreds. Analysis of DNA polymorphisms in the fibrinogen alpha-chain gene locus (TCTT repeat in intron 3, Rsal site in exon 5, and Taql site in the 3' flanking region of the gene) showed the haplotype B5-Rsal(+)-Taql(-) for the Val 526 mutant gene in both kindreds studied here, as well as in two kindreds previously described (J Clin Invest 1994; 93:731). The fibrinogen alpha-chain gene mutation (Val 526) is the genetic defect responsible for hereditary renal amyloidosis in these two kindreds, and the mutant genes in the Val 526 kindreds may have been derived from a single founder.      

Commentary on “A novel treatment strategy for newly diagnosed high-grade T1 bladder cancer: Gemcitabine and cisplatin adjuvant chemotherapy—A single-institution experience.”

Background

Management of high-grade T1 (formerly T1G3) bladder cancer continues to be controversial. Should patients with T1G3 bladder cancer have an immediate radical cystectomy or should they receive intravesical bacillus Calmette-Guérin preserving bladder? Gemcitabine and cisplatin (GC) adjuvant chemotherapy may help to strike a balance between intravesical and early cystectomy. For purposes of this study, we continue to refer high-grade T1 lesion as “T1G3.”

胶原海绵复合新生大鼠原代心肌细胞构建工程化心肌组织

目的:探索以胶原海绵为支架、新生大鼠原代心肌细胞为种子细胞,于体外构建工程化心肌组织的方法。方法:实验于2005-12/2006-11在解放军第四军医大学西京医院心内科实验室完成。Ⅰ型胶原海绵剪切成方形片状(2.0cm×1.4cm×0.2cm),经60Co照射消毒,于DMEM培养液中水化1h左右。另取1d龄SD大鼠心脏,剪成小碎块,然后用2.5g/L胰蛋白酶于37℃中消化,吸取上清至含胎牛血清的DMEM中,重复消化四五次,用差速贴壁法除去大部分成纤维细胞,将细胞沉淀用DMEM培养液以2×109L-1的密度悬浮备用。将上述的心肌细胞悬液1mL缓慢滴注于玻璃模型中的胶原海绵上,然后置于细胞培养中培养。肉眼及显微镜主要观察工程化心肌组织在培养期间的自发收缩情况,包括收缩的部位、强度、频率、一致性以及收缩随时间变化的情况。苏木精-伊红染色观察工程化心肌组织内胶原纤维的变化,细胞形态,胞核的形状及细胞之间的连接。免疫组织化学染色和透射电镜观察工程化心肌组织片的形态和功能。结果:①细胞接种于胶原海绵上1d后,细胞/胶原复合物的凝胶化过程基本完毕,体积保持恒定,维持至培养结束,第3天细胞/胶原复合物局部出现点片状自发收缩,第5天整个细胞/胶原复合物出现同步化自发收缩,收缩频率61～199次/min。2周后37.5%的工程化心肌组织的自发收缩活动减弱,但75%的工程化心肌组织的自发收缩活动持续至培养结束。②苏木精-伊红染色、免疫组织化学染色和透射电子显微镜显示,工程化心肌组织内细胞间连接广泛存在,细胞多呈纵向分布,胞核呈长圆形,胞浆内α-肌节肌动蛋白阳性,胞内肌原纤维排列整齐,可见到心肌特异性的肌小节结构和Z线,多数细胞具有分化的心肌细胞表型。结论:用新生大鼠原代心肌细胞为种子细胞、以Ⅰ型胶原海绵为支架材料,构建出的工程化心肌组织,于体外可长时间持续自发收缩,该细胞/胶原复合物的形态结构与生理功能均类似于成熟大鼠心肌组织。     

言语流畅的测量及其神经生理基础

目的:介绍言语流畅性的测量工具,考察言语流畅性与记忆之间的关系,探讨言语流畅性的神经生理基础。资料来源:应用计算机检索Sciencedirect 1975-07/2005-05中与言语流畅性相关的文献,检索词为“verbal fluency”,限定文章语言种类为英文。资料选择:对资料进行初审,选取文章标题中含有“verbal fluency”的英文文献。纳入标准:①对言语流畅性测量任务的研究。②对言语流畅性与记忆的研究。③对言语流畅性神经生理基础的研究。资料提炼:共收集到651篇关于言语流畅性的文章,纳入30篇用于本综述。资料综合:测量言语流畅性的任务主要分为简单任务(包括音位流畅性任务和语义流畅性任务)与复杂任务(包括可能工作、替代使用、草拟、推断、物品制作以及新的使用方法等)。言语流畅性任务的完成主要涉及到记忆搜索的过程,另外,精细组织的语义网络系统对于言语流畅性任务来说也是非常重要的。言语流畅性的神经生理基础包括了总体基础,前额叶,海马以及大脑两半球的不对称性。结论:言语流畅性作为人类语言基本技能的一项指标,对语言认知研究具有极其重要的意义。它不仅是神经心理研究的一项非常重要的检测和诊断指标,也是一种研究人类心理的非常有用的研究工具,是探索人类语言与认知、创造之迷进而探索人类意识之迷的钥匙。