Haemophilia A and haemophilia B: molecular insights

This review focuses on selected areas that should interest both the scientist and the clinician alike: polymorphisms within the factor VIII and factor IX genes, their linkage, and their ethnic variation; a general assessment of mutations within both genes and a detailed inspection of the molecular pathology of certain mutations to illustrate the diverse cause–effect relations that exist; a summary of current knowledge on molecular aspects of inhibitor production; and an introduction to the new areas of factor VIII and factor IX catabolism. An appendix defining various terms encountered in the molecular genetics of the haemophilias is included, together with an appendix providing accession numbers and locus identification links for accessing gene and sequence information in the international nucleic acid databases.     

Comparison of gas-liquid chromatography and EMIT assay for serum valproic acid

We describe a simple direct extraction method for the gas-liquid chromatography determination of serum valproic acid. The working range for the assay is 2-180 mg/L and our within-run precision was 5.8 and 4.3% at the 40 and 90 mg/L concentrations respectively. Hemolyzed and lipemic sera as well as samples from patients with hyperbilirubinemia and from patients with decreased renal function were put through the assay and no interfering peaks were noted. Interference occurred when teflon-lined screw caps were used during the extraction step. The method was proven to be accurate by linear regression analysis of samples containing weighed-in amounts of valproic acid. The above assay was compared to an enzyme immunoassay technique (EMIT). The working range for the latter is 10-150 mg/L and the with-run precision was 10.8 and 5.9% and 90 mg/L concentration respectively. Samples were run by both the gas-liquid chromatograph and enzyme immunoassay methods and gave very similar results over the range 16-139 mg/L.     

重组结核分支杆菌16000蛋白抗原(rPA16)的免疫原性研究

目的：研究结核分枝杆菌重组16000蛋白（rPA16)诱发家兔产生抗体的能力并评价其血清学诊断的价值。方法：家兔按注射的抗原及佐剂的不同分组，皮下多点注射分别进行免疫，6次免疫后采血，双向琼脂扩散试验和酶联免疫吸附试验（ELISA）跟踪兔血清抗体效价的时间效应并检测兔血清与所试各类分支杆菌PPD有无交叉反应；同时检测rPA16与各分支杆菌PPD的免疫羊血清的反应，并用rPA16抗原检测人血标本，分析该抗原的临床检测的敏感性和特异性。结果：兔血清抗体效价结果显示：5个月后，加佐剂的抗原组的兔免疫血清仍能检测到抗体，不加佐剂组2个月后血清抗体检测即为阴性，ELISA检测兔免疫血清比双向琼脂扩散方法要更敏感，rPA16抗原加佐剂引起的兔血清抗体滴度最高可达1：6400，不加佐剂组的血清抗体滴度仅为1：400，ELISA法分析抗rPA16的兔血清的特异性表明其特异笥较好，仅与牛、人型结核分支杆菌的PPD呈阳性反应，与其他所试分支杆菌PPD反应均为阴性，同时又分析了rPA16抗原的特异性，与人型PPD相比，该抗原只与所试各类分支杆菌，BCG、牛分支杆菌免疫的血清呈阳性反应，而人型PPD则基本上与结核分支杆菌免疫的血清均呈阳性反应，ELISA检测血清标本结果：肺结核病组中rPA16和PPD检测敏感性分别为64.8%和75.2%；健康组，卡介苗接种阳转健康组，rPA16和PPD检测特异性分别为96.2%,88.5%和94.3%,66.7%。结论：rPA16具有较强的免疫原性和一定的特异性，可能成为结核病血清学诊断试剂之一。     

PTEN基因蛋白在胆囊癌组织中表达的研究

目的 :探讨抑癌基因PTEN蛋白在胆囊癌组织中的表达情况及其临床病理意义。方法 :应用免疫组化对 2 8例胆囊腺癌 ,4例胆囊鳞癌及其相应癌旁组织中PTEN蛋白进行检测 ,并结合患者的临床病理资料进行分析。结果 :PTEN蛋白在 32例癌旁组织中全都呈阳性表达 ,而癌组织中PTEN阳性表达率仅 34.4 %。胆囊癌组织中PTEN蛋白阳性表达与患者性别、年龄及组织类型无明显相关性 ,但与组织分化程度明显相关。结论 :PTEN蛋白在胆囊癌组织中阳性表达率较低 ,说明该基因失活可能在胆囊癌发生发展中起重要作用。PTEN阳性表达对胆囊癌患者的预后有一定意义     

钾通道阻断剂对低氧/复氧诱导的培养海马神经元死亡的防护作用

目的研究钾通道阻断剂对单纯低氧/复氧诱导的培养海马神经元死亡的防护作用。方法培养8 d的海马神经元置于低氧环境(95% N2/5% CO2)6 h,复氧再培养直至72 h,复氧后0.5 h培养液内分别给予不同钾通道阻断剂,用细胞计数及MTT比色法检测神经元死亡情况。结果低氧/复氧诱导培养海马神经元出现迟发性死亡;四乙铵以剂量依赖性的方式防护低氧/复氧诱导的培养海马神经元免于死亡;大电导钙激活钾通道(BK通道)阻断剂iberiotoxin(IbTX)可完全消除低氧/复氧诱导的神经元死亡(P<0.001);A型钾通道阻断剂4-氨基吡啶不能防护低氧/复氧诱导的神经元免于死亡(P>0.05)。结论钾通道阻断剂四乙铵和IbTX能防护低氧/复氧诱导的培养海马神经元免于死亡,提示某些类型的钾通道尤其是BK通道活动增强可能参与了低氧/复氧诱导的培养海马神经元死亡。     

MMP-9和TIMP-1表达失衡与乳腺癌浸润、转移的相关性

目的 研究乳腺癌组织中基质金属蛋白酶（MMP－9）和金属蛋白酶组织抑制因子（TIMP－1）的表达变化与乳腺癌生物学行为及淋巴结转移的关系。方法 应用SP免疫组织化学方法检测85例乳腺癌组织MMP－9，TIMP－1及细胞增殖核抗原ki-67的表达情况。结果 MMP-9阳性染色率90．59％，MMP－9阳性表达与肿瘤浸润，淋巴结转移，ki-67指数及TNM分期呈正相关（Pearson列联系数分别为P=0.03,P=0.02,P=0.004和P＝0．0000，P＜0．05，0．01。TIMP－1阳性染色率为78．82％，TIMP－1阳性表达与肿瘤浸润，淋巴结转移及TNM分期浸润，转移及ki-67指数显著相关（P＜0．05，P＜0．01，P＜0．001）。结论 MMP－9和TIMP－1表达失衡与乳腺癌浸润及淋巴结转移密切相关。     

Pharmacokinetics, metabolism and renal excretion of sulfatroxazole and its 5-hydroxy- and N4-acetyl-metabolites in man

Hydroxylation is the predominant pathway of metabolism for sulfatroxazole in the body, accounting for 70 per cent of the dose. Fifteen per cent of the dose is acetylated unimodally and 10 per cent is excreted unchanged. The half-lives of sulfatroxazole and its metabolites 5-hydroxysulfatroxazole and N4-acetylsulfatroxazole are approximately 22 h after administration of sulfatroxazole. N4-acetylsulfatroxazole, taken as parent drug, is eliminated by renal excretion (92 per cent of the dose). The initial elimination half-life of N4-acetylsulfatroxazole is 4.5 h, which later increases to 70 h as the result of the acetylation-deacetylation equilibrium. Probenecid inhibits the renal excretion of the metabolites 5-hydroxy- and N4-acetylsulfatroxazole. Inhibition of the N4-acetyl metabolite favours the deacetylation, which results in an increase of the T 1/2 of sulfatroxazole from 20 to 30 h. The protein binding value of sulfatroxazole is 84 per cent, that of N4-acetylsulfatroxazole is 37 per cent. Sulfatroxazole is excreted renally by passive processes, while the metabolites are excreted by both passive and active processes.     

稳定性核素测定大鼠小肠蛋白质合成

目的：建立稳定性核素（[L-^15N]亮氨酸）测定大鼠小肠蛋白质合成率的方法。方法：分别测定静脉注射相同剂量[L-^15N]亮氨酸不同时相的大鼠小肠^15N丰度及不同剂量[L-^15N]亮氨酸同一时相的大鼠小肠^15N丰度。结果：大鼠小肠游离氨基酸池中^15N核素丰度在注射后0．5h内呈线性上升并达高峰，维持4h后缓慢下降，小肠蛋白质中的^15N丰度0．5h至12h基本维持不变；随着注射剂量的增加，大鼠小肠蛋白质分数合成率（FSR）亦增加，当[L-^15N]亮氨酸剂量在1．0mmol／kg以上，FSR并不随施加[L15N]亮氨酸剂量的加大而增加。结论：在进行大鼠小肠蛋白质合成率测定时，一次性静脉注射的测量最佳时限为0．5h，剂量为1．0mmol／kg。     

CLINICAL OBSERVATION ON THERAPEUTIC EFFECTS OF COMBINED TREATMENT OF NONINSULIN DEPENDENT DIABETES

Thetherapeuticeffectofacupuncturefordia betesmellitus(DM)hasbeenconfirmedbyalarge numberofclinicalinvestigations.Itisasubjectfor acupuncturiststoconsiderandsearchforatherapy providingbettertherapeuticeffectsbywayofproper differentiationofsyndromesandcombinationofacu points.Basedonclinicalexperiencegainedformore than30years,theauthorsofthepresentpaperhave graduallyformedaneffectivecombinedtherapyfor treatingtypeⅡDM(noninsulindependentDM)and itsvariouscomplications.Thereportisasfollows.1CLI…