Rh phenotype prediction by DNA typing and its application to practice

The complexity of the RHD and RHCE genes, which is the greatest of all blood group systems, confounds analysis at the molecular level. RH DNA typing was introduced in 1993 and has been applied to prenatal testing. PCR-SSP analysis covering multiple polymorphisms was recently introduced for the screening and initial characterization of partial D. Our objective is to summarize the accrued knowledge relevant to the approaches to Rh phenotype prediction by DNA typing, their possible applications beyond research laboratories and their limitations. The procedures, results and problems encountered are highly detailed. It is recommended that DNA typing comprises an analysis of more than one polymorphism. We discuss future directions and propose a piecemeal approach to improve reliability and cost-efficiency of blood group genotyping that may eventually replace the prevalent serology-based techniques even for many routine tasks. Transfusion medicine is in the unique position of being able to utilize the most extensive phenotype databases available to check and develop genotyping strategies.     

易善复加丹参治疗病毒性肝炎合并脂肪肝疗效观察

目的　评价易善复加丹参在病毒性肝炎合并脂肪肝的临床疗效。方法　选择病毒性肝炎合并脂肪肝 98例 ,随机分成治疗组和对照组 ,治疗组 68例 ,应用易善复加丹参治疗 ,疗程 3个月。结果　治疗组血脂 (TC、TG)、肝功能 (ALT、γ -GT)、B超脂肪肝图象及临床症状等项目的改善均明显优于对照组 (P <0 .0 1～ 0 .0 5) ,且无明显副作用。结论　易善复加丹参治疗病毒性肝炎合并脂肪肝对促进肝脏脂肪代谢、降低血脂、修复损伤的肝细胞 ,且阻止或改善肝纤维化均有明显的功效     

肺肿瘤，肺心病患者红细胞免疫功能的观察

本文用红细胞酵母菌花环率方法对３６例肺肿瘤、５５例肺心病患者红细胞免疫功能和９４例正常健康人红细胞免疫功能进行检测，结果表明：肺肿瘤、肺心病急性加重期患者红细胞免疫功能明显低于健康人，肺心病缓解期患者红细胞免疫功能有所恢复，肺心病合并多系统脏器功能衰竭患者红细胞免疫功能进一步降低。     

几丁糖预防腹膜粘连抑制胶原基因表达的实验研究

目的 :探讨几丁糖在胶原基因表达及预防大鼠腹膜粘连形成中的作用。方法 :制作大鼠回肠浆膜均一缺损的腹膜粘连模型 ,关腹前注入乳酸Ringer液或几丁糖 ,术后 14天进行粘连评分。采用半定量逆转录 -聚合酶链反应 (RT PCR)方法测定各粘连组织标本Ⅰ型胶原基因mRNA的表达 ;采用苦味酸天狼猩红染色 -偏振光法及计算机图像分析系统来观察和分析Ⅰ型和Ⅲ型胶原纤维在粘连组织中的分布及染色表达面积。结果 :与正常腹膜和乳酸Ringer’s液组相比 ,几丁糖组大鼠的粘连显著减轻 ,Ⅰ型胶原基因mRNA的表达减弱 ,Ⅰ型胶原形成减少 (P <0 .0 1) ,Ⅲ型胶原在各组间无差别 (P >0 .0 5 )。结论 :胶原基因的过度表达可能在粘连形成过程中有重要的作用 ,几丁糖能抑制Ⅰ型胶原基因的表达 ,减少Ⅰ型胶原的合成 ,从而减轻粘连的程度     

一例~(192)Ir极不均匀外照射局部极重度放射损伤的临床观察

目的对１例１９２Ｉｒ极不均匀外照射局部极重度放射损伤病人进行临床观察。方法系统观察了病变的临床过程和应用红外线热成像技术测定了损伤部位的温度变化。结果照射后２小时出现肢体麻木、抽搐，最早照后４小时出现红斑、肿胀，５４小时出现水疱，第５天出现坏死和剧痛，最晚出现红斑、肿胀是照射后４１天，４７天出现水疱和糜烂创面。红外线热成像显示：损伤早期温度升高，水疱、坏死区和损伤后期温度降低，温度升高越早，损伤越重，温度改变区域与损伤范围相一致。结论大剂量极不均匀外照射放射损伤的初期反应早，放射病与局部损伤相互加重病情，症状先后不一，轻重不等，致残率高和疼痛剧烈，红外线热成像温度变化可以作为临床诊断的指标之一。     

杆状病毒介导LacZ基因转染体外培养的人虹膜色素上皮细胞

目的：研究重组杆状病毒(baculovinus)载体介导β—半乳糖苷酶基因(LacZ)对体外培养虹膜色素上皮(IPE)细胞的转染和表达情况。方法：将质粒pCMV—β中的CMV—LacZ表达盒插入杆状病毒表达栽体pFast BacI的Eco RI／Hind Ⅲ位点，构建重组质粒pBac—β，并利用Bac—to—Bac表达系统在Sf9细胞中产生出重组杆状病毒BV—β。将MOI为200的BV—β感染经酶消化加显微法分离培养的IPE细胞，24h后进行X-gal染色。在显微镜下观察IPE中LacZ表达阳性的情况，记录阳性细胞所占的百分比。结果：限制性酶切分析及测序结果表明，我们已成功构建表达质粒pBac—β及重组杆状病毒BV—β。X-gal染色证实杆状病毒介导LacZ在细胞中的表达呈蓝色，弥漫于整个胞浆。感染后24h时表达阳性率达85％。结论：重组杆状病毒可有效介导报告基因在体外培养的IPE细胞中表达，为利用杆状病毒构建可供移植的基因工程化虹膜色素上皮细胞提供了实验依据。     

Evidence for delayed cytotoxicity effects following exposure of rat hepatoma-derived Fa32 cells: implications for predicting human acute toxicity

The delayed cytotoxicity of the Multicentre Evaluation of In vitro Cytotoxicity (MEIC) reference chemicals was investigated in rat hepatoma-derived Fa32 cells. The cells were treated for 24 h with the test chemicals, and were than further cultured for 5 days in normal culture medium. The cytotoxicity was measured by the neutral red uptake inhibition, and the results were quantified by determining the NI50del. This is the concentration of test compound required to decrease the neutral red uptake with 50% compared with control cells. The results were compared with the acute NI50, the corresponding value measured immediately after 24 h treatment of the cells. On a total of 44 chemicals, nine showed delayed cytotoxicity (NI50del lower than or equal to NI50), 11 a probably delayed, and 24 no delayed cytotoxicity (NI50del more than 1.5×NI50). When the NI50del was compared with human toxicity, a correlation coefficient r²⁼0.761 was obtained. For the same series of 44 chemicals this correlation was clearly higher than that for human hepatoma-derived Hep G2 cells (r²=0.695). The Hep G2 assay was the best acute in vitro assay for the prediction of human toxicity within the MEIC study. Consequently, the delayed cytotoxicity assay on cultured Fa32 cells has the best prediction value so far obtained for the human toxicity.     

Luminal transport step of para-aminohippurate (PAH): transport from PAH-loaded proximal tubular cells into the tubular lumen of the rat kidney in vivo

 Proximal tubular cells were loaded for 10 s with [³H]para-aminohippurate ([³H]PAH) by microperfusing the peritubular capillaries with Ringer solution containing 0.05 mmol/l PAH. Immediately thereafter [³H]PAH influx from cells into a column of equilibrium solution injected into the oil-filled tubular lumen was measured by re-aspirating the fluid after 1–10 s of contact time. The rise of luminal PAH concentration within 2 s of contact time was almost linear, reaching a luminal / capillary concentation ratio of 1.6 after 2 s and of 3.2 after 5 s. The 2-s PAH concentration ratio was not changed when different manoeuvres were applied to depolarize proximal tubular cells. Also, the 2-s PAH concentration ratio was not influenced by varying the luminal pH from 6.0 to 8.0 or the luminal Cl^–concentration from zero to 134 mmol/l or when either 5 mmol/l urate or 25 mmol/l lactate was in the luminal perfusate. A decrease in the 2-s PAH concentration ratio, i.e. trans-inhibition, was observed when 25 or 50 mmol/l HCO₃ ^–(–50%) was in the luminal perfusate. Trans-inhibition was also seen with 5 mmol/l of the following substituted benzoates: 2-hydroxy-benzoate (–58%), 2-methoxy-benzoate (–46%), 2-hydroxy-benzoate-acetyl ester (–36%), 2-hydroxy-3,5-dinitro-benzoate (–48%), 3,5-dichloro-benzoate (–49%), and 2,3,5-trichloro-benzoate (–45%). No effect was seen with benzoate, 3-hydroxy-benzoate, 2-chloro-benzoate, 2-nitro-benzoate, 2,5-dinitro-benzoate, 3-sulfamoyl-benzoate and 4-sulfamoyl-benzoate. However, analogues of the latter two compounds possessing two additional side groups, such as furosemide and piretanide, or a hydrophobic moiety, such as probenecid, were inhibitory (by –62, –41 and –49% respectively). Phenoxyacetate had no effect; however, it inhibited if in addition it had three chloro groups, as in 2,4,5-trichlorophenoxyacetate (–71%) or a hydrophobic carbamoyl side group, as in mersalylic acid (salyrgan, –75%). Benzene-sulfonate trans-inhibited (–33%), as did phenolsulfonphthalein (phenol red, –39%) and sulfofluorescein (–55%). However, the trans-inhibitory effect of the corresponding carboxy-compounds was absent (phenolphthalein) or weaker (fluorescein, –42%). The trans-inhibitory effect of the uricosurics ethacrynic acid (–53%), tienilic acid (–55%) indacrinone (–72%) and benzbromarone (–42%) could be attributed to two chloro or bromo side groups on the benzene ring. Other trans-inhibiting uricosuric substances were indomethacin (–42%), sulfinpyrazone (–38%), losartan (–80%) its metabolite EXP 3174 (–55%), and AA 193 (–65%). These organic acids, with pKa values between 2.8 and 4.9, possess chloro and sulfin groups, as well as heterocyclic 5-ring and hydrophobic ring or chain areas. No significant effect was seen with 5 mmol/l PAH, 2-oxo-glutarate, DIDS, cGMP, prostaglandin E₂, cortisol, benzylamiloride, pyrazinoic acid and 25 mmol/l lactate. Our data indicate that in situ the secretory luminal PAH transport proceeds in a non-rheogenic fashion, per exclusionem by anion exchange. The observed trans-inhibition of PAH secretion seems to correlate with the affinity for the luminal PAH transporter and, for uricosuric substances, with their uricosuric potency. Received: 15 October 1996 / Received after revision: 17 December 1996 / Accepted: 18 December 1996     

Elevated anti-alginate serum titers in patients with acute cholangitis and gallstones imbedded withPseudomonas aeruginosa

Electron microscopy and bacteriological culture revealed viable bacteria covered with a glycocalyx (biofilm) in choledochal stones recovered from two patients with acute cholangitis. On the cut surface of the choledochal stones, the cholesterol stone component was surrounded with a layer of brown pigment stone. In each case, bacterial culture of the choledochal stone recoveredPseudomonas aeruginosa. Since alginate is the main component of the glycocalyx produced byP. aeruginosa, serum IgM, IgG and IgA anti-alginate antibodies were measured in each patient. The present study is the first to demonstrate acute and transient IgM seroconversion to alginate in cases of acute cholangitis. In one case, the elevation of anti-alginate IgM preceded the elevation of anti-alginate IgG. The authors propose that the bacterial glycocalyx may play a significant role in acute cholangitis.     

红色素基因RFLP及其与情感性精神病关系的研究

以红色素基因全长ｃＤＮＡ作探针，２０例正常人，２６例双相型情感性精神病患者进行ＲＦＬＰ分析。ＳａｃＩ酶切显示：部分正常人和部分患者基因组ＤＮＡ产生的７．５ｋｂ，４．８ｋｂ，４．５ｋｂ呈多态性改变的片段。正常人具多态改变的占４５％，患者为４６．２％，按Ｘ染色体数计，正常人出现７．５ｋｂ限制性片段的频率为１４．３％，患者为３２．４％，但两者差异不显著。