Conformational study of glycopeptides

The conformational feature has been studied by n.m.r. spectroscopy on the compounds, Boc-Asn-NHMe, Boc-Asn-Gly-NHMe, Boc-Gly-Asn-NHMe, and their glycosylated derivatives. From the temperature dependence of the amide proton chemical shifts and vicinal coupling constants, little change was confirmed in the peptide conformation upon N-glycosylation. There is no particular intramolecular interaction between the peptide and carbohydrate moieties. Boc-Asn-Gly-NHMe takes, to some extent, a folded structure with a hydrogen bond involving the amide proton of N-methylamide group. This backbone conformation is also preferable in the corresponding glycopeptide.     

A Case of Rectal Carcinoma with Elevated Serum Alpha-Fetoprotein

A gradual increase in serum alpha-fetoprotein (AFP) had beenobserved in a patient with liver cirrhosis and the complicationof rectal carcinoma was disclosed 18 months later. The concentrationof serum AFP decreased rapidly after removal of the tumor andreached a normal value within a month. The level of serum AFP has remained normal and the patient hasbeen well since the operation. There has been no clinical evidenceof the existence of a liver tumor and the production of AFPby the rectal carcinoma cells was suggested.     

The role of pretreatment squamous cell carcinoma antigen level in locally advanced squamous cell carcinoma of the uterine cervix treated by radiotherapy

The purpose of this study was to determine the pretreatment serum squamous cell carcinoma antigen (SCC-ag) level as a generally applicable measurement in predicting and estimating the treatment outcome of patients with locally advanced SCC of the cervix. Three hundred fifty-two patients with stage IIB-IVA SCC of the cervix were managed with both external irradiation and high-dose rate intracavitary brachytherapy. A significantly higher median SCC-ag was seen in association with increasing stage, tumor size, and lymph node involvement. The difference in disease-free survival (DFS) between stages IIB and III patients was not statistically significant with SCC-ag level <2 ng/mL. In multivariate analysis, median SCC-ag level (> or =6.0 ng/mL) and lymph node metastases had significant independent effects on absolute survival and DFS. A direct linear relationship (y=-2.932x+ 84.896) existed between the median SCC-ag of groups distributed by pretreatment prognostic factors and the 5-year DFS rate. The 5-year DFS rate as a function of SCC-ag level defined by cervix size, lymph node status, and hydronephrosis was obtained from a formula combining risk scores and the baseline survival function. From the obtained formulas, we can objectively estimate the treatment outcome in patients with locally advanced squamous cell cervical cancer.     

Roles of two allelic variants (Arg144Cys and Ile359Leu) of cytochrome P4502C9 in the oxidation of tolbutamide and warfarin by human liver microsomes

1. Tolbutamide methyl hydroxylation and racemic warfarin 7-hydroxylation activities were determined in liver microsomes of 39 Japanese and 45 Caucasians genotyped for the cytochrome P450 (P450 or CYP) 2C9 gene into three groups, namely the wild-type (Arg144·Ile359), and two heterozygous Cys allele (Cys144·Ile359) and Leu allele (Arg144·Leu359) variants. 2. Good correlations were found between tolbutamide methyl hydroxylation and racemic warfarin 7-hydroxylation activities in liver microsomes of Japanese and Caucasians. Humans with the Cys allele CYP2C9 variant, which was detected in 22% of Caucasians, were found to have similar catalytic rates to those of the wild-type in the oxidations of tolbutamide and racemic warfarin, whereas humans with the Leu allele, which was detected in 8% Japanese and 7% Caucasian samples, had lower catalytic rates than those of other two groups. 3. The rates of 6- and 7-hydroxylation of racemic warfarin were correlated well with those of S-warfarin, but not R-warfarin, in human liver microsomes. 4. Both human liver microsomes and recombinant CYP2C9 catalysed 7-hydroxylation of S-warfarin more extensively than those of R-warfarin. K_m's for the 7-hydroxylation of S-warfarin were not very different in liver microsomes of humans with these three genotypes. Anti-CYP2C9 antibodies and sulphaphenazole inhibited the 6- and 7- hydroxylation of S-warfarin, but not R-warfarin,by > 90% and the methyl hydroxylation of tolbutamide by about 50%. 5. These results suggest that humans with Leu allele of CYP2C9 have lower V_max's for S-warfarin 7-hydroxylation and tolbutamide methyl hydroxylation than those with wildtype and Cys allele CYP2C9, although the K_m's are not very different in liver microsomes m of these three groups of humans. R-warfarin hydroxylation may be catalysed by P450 enzymes other than CYP2C9 in man.     

组织蛋白酶K与高血压性心脏病心室肌重塑及纤维化关系的初步探讨

目的:探讨组织蛋白酶K(cathepsin K,Cat K)在高血压性心脏病心室重塑发生发展中的表达及其活性与心肌纤维化的关系.方法:7周龄雄性Dahl盐敏感(DS)大鼠分别给予高负荷食盐(8%)或正常负荷食盐(0.3%)19周,建立高血压性心脏痛心衰(H-HF)模型和对照组(19W-C).另设人心肌活检标本作研究对象(H-HF患者10例和对照组6例).用RT-PCR和免疫组化分别检测Cat K mRNA及蛋白表达,酶谱法测定大鼠心肌组织中的组织蛋白酶活性,心脏石蜡切片Azan染色法分析左心室肌间质和血管周围纤维化程度,细胞荧光染色法分析白介素-β1(IL-β1)对培养心肌细胞的Cat K的表达及其胶原蛋白分解能的影响.结果:H-HF组大鼠和患者心肌中的Cat K mRNA的表达显著高于19W-C组和对照组人(P<0.01).免疫组化法和in situ法分析显示增高的Cat K mRNA和蛋白主要表达于心肌细胞.H-HF组大鼠心肌组织蛋白酶依赖性胶原蛋白分解能显著高于19W-C组.H-HF组大鼠心肌中IL-β1表达也在显著增高.IL-β1刺激心肌细胞的Cat K蛋白及其胶原蛋白分解活性.心肌中的Cat K mRNA表达与左心室肌纤维化面积呈显著正相关(r=0.68,P<0.01).结论:在心肌中Cat K表达及分泌受IL-β1的调控,并参与高血压性心脏病的发生、发展过程,提示心肌Cat K在H-HF的心肌重塑及纤维化发展中起重要作用.     

Intra‐articular injection of mesenchymal stem cells expressing coagulation factor ameliorates hemophilic arthropathy in factor VIII‐deficient mice

Summary. Background: Transplantation of cells overexpressing a target protein represents a viable gene therapeutic approach for treating hemophilia. Here, we focused on the use of autologous mesenchymal stem cells (MSCs) expressing coagulation factor for the treatment of coagulation factor VIII (FVIII) deficiency in mice. Methods and Results: Analysis of luciferase gene constructs driven by different promoters revealed that the plasminogen activator inhibitor‐1 (PAI‐1) gene promoter coupled with the cytomegalovirus promoter enhancer region was one of the most effective promoters for producing the target protein. MSCs transduced with the simian immunodeficiency virus (SIV) vector containing the FVIII gene driven by the PAI‐1 promoter expressed FVIII for several months, and this expression was maintained after multiple mesenchymal lineage differentiation. Although intravenous injection of cell supernatant derived from MSCs transduced with an SIV vector containing the FVIII gene driven by the PAI‐1 promoter significantly increased plasma FVIII levels, subcutaneous implantation of the MSCs resulted in a transient and weak increase in plasma FVIII levels in FVIII‐deficient mice. Interestingly, intra‐articular injection of the transduced MSCs significantly ameliorated the hemarthrosis and hemophilic arthropathy induced by knee joint needle puncture in FVIII‐deficient mice. The therapeutic effects of a single intra‐articular injection of transduced MSCs to inhibit joint bleeding persisted for at least 8 weeks after administration. Conclusions: MSCs provide a promising autologous cell source for the production of coagulation factor. Intra‐articular injection of MSCs expressing coagulation factor may offer an attractive treatment approach for hemophilic arthropathy.     

中枢免疫耐受缺损模型的构建

目的构建基于shRNA介导的中枢免疫耐受缺损模型。方法分离妊娠后13.5 d的胎鼠胸腺,除去淋巴细胞,获得原代胸腺基质细胞,利用慢病毒将小鼠TRAF6基因特异性shRNA慢病毒质粒(LV-T6-shRNA)导入原代胸腺基质细胞中,重新聚集这些胸腺基质细胞得到LV-T6-shRNA重塑胸腺,将此重塑胸腺移植入无胸腺的雌性小鼠肾脏荚膜下,饲养8周。结果慢病毒可有效转导LV-T6-shRNA慢病毒质粒入原代胸腺髓质上皮细胞中;移植LV-T6-shRNA重塑胸腺的小鼠饲养8周后,胸腺明显较小,成熟的胸腺髓质上皮细胞减少,脾脏肿大,脾脏中活化的T淋巴细胞增多,肺脏中出现淋巴细胞浸润等现象,这些表现型与TRAF6-/-小鼠的表现型相似。结论中枢免疫耐受缺损模型构建成功。     

孤立性髂动脉瘤36例的临床诊断和治疗

目的提高对孤立性髂动脉瘤（isolated iliae artery aneurysm,IIAA）的认识,总结临床诊断和治疗经验。方法回顾性分析1983年1月至2006年3月期间收治的36例IIAA的临床资料。无症状患者17例,有症状患者19例。择期手术治疗33例,急诊手术3例。治疗措施主要包括动脉瘤切除和动脉重建术。辅助手术包括输尿管松解术和输尿管切断再缝合。结果择期手术术后发生切口延期愈合1例,臀肌跛行2例,下肢运动和排尿障碍1例,均经药物保守治疗,症状减轻和消失,其余患者均康复良好出院。急诊手术3例于术后死亡。结论IIAA临床罕见,破裂出血死亡率高,早期诊断和手术治疗是降低死亡率的关键。微创血管腔内治疗术是值得推荐应用的方法。