Preparation of protected peptidyl thioester intermediates for native chemical ligation by Nα‐9‐fluorenylmethoxycarbonyl (Fmoc) chemistry: considerations of side‐chain and backbone anchoring strategies,and compatible protection for N‐terminal cysteine*,†

Abstract: Native chemical ligation has proven to be a powerful method for the synthesis of small proteins and the semisynthesis of larger ones. The essential synthetic intermediates, which are C‐terminal peptide thioesters, cannot survive the repetitive piperidine deprotection steps of N^α‐9‐fluorenylmethoxycarbonyl (Fmoc) chemistry. Therefore, peptide scientists who prefer to not use N^α‐t‐butyloxycarbonyl (Boc) chemistry need to adopt more esoteric strategies and tactics in order to integrate ligation approaches with Fmoc chemistry. In the present work, side‐chain and backbone anchoring strategies have been used to prepare the required suitably (partially) protected and/or activated peptide intermediates spanning the length of bovine pancreatic trypsin inhibitor (BPTI). Three separate strategies for managing the critical N‐terminal cysteine residue have been developed: (i) incorporation of N^α‐9‐fluorenylmethoxycarbonyl‐S‐(N‐methyl‐N‐phenylcarbamoyl)sulfenylcysteine [Fmoc‐Cys(Snm)‐OH], allowing creation of an otherwise fully protected resin‐bound intermediate with N‐terminal free Cys; (ii) incorporation of N^α‐9‐fluorenylmethoxycarbonyl‐S‐triphenylmethylcysteine [Fmoc‐Cys(Trt)‐OH], generating a stable Fmoc‐Cys(H)‐peptide upon acidolytic cleavage; and (iii) incorporation of N^α‐t‐butyloxycarbonyl‐S‐fluorenylmethylcysteine [Boc‐Cys(Fm)‐OH], generating a stable H‐Cys(Fm)‐peptide upon cleavage. In separate stages of these strategies, thioesters are established at the C‐termini by selective deprotection and coupling steps carried out while peptides remain bound to the supports. Pilot native chemical ligations were pursued directly on‐resin, as well as in solution after cleavage/purification.     

门静脉主干缩窄法制备SD大鼠门静脉高压症模型时最佳口径的探讨

目的 探讨缩窄门静脉主干法制备SD大鼠门静脉高压症模型时的最佳缩窄口径.方法 SD大鼠70只,随机分为正常组和6个实验组,每组各10只.正常组行假手术.各实验组分别按照5、6、7、8、9、12号针头的缩窄口径行门静脉主干缩窄术.观察各组大鼠术后累积死亡率,术后状态,术前、术后即刻及术后2周时的门静脉压力,术后2周时的食管组织学变化和脾指数.结果 5、6、7、8、9、12号针头缩窄组术后3 d时大鼠的累积死亡率分别为100%、80%、70%、20%、10%、0%,与缩窄程度正相关.8、9、12号组的大鼠存活状态明显好于5、6、7号组.5、6、7、8、9、12号组术后即刻门静脉压力分别为:(5.836±0.275)、(4.557±0.419)、(3.856±0.576)、(3.343±0.433)、(2.708±0.309)、(1.957±0.358)kPa,7、8、9、12号组术后2周时门静脉压力分别为:(2.163±0.424)、(1.956±0.172)、(1.841±0.202)、(1.232±0.154)kPa,均较正常(0.881±0.165)kPa显著升高(P＜0.05).术后2周,7、8、9、12号组大鼠食管下段黏膜下层平均血管数目分别为:(3.94±0.83)、(3.58±0.63)、(3.14±0.64)、(2.02±0.62)个,与正常组(1.65±0.62)个比较,除12号组外均有增多(P＜0.01);固有层平均血管数目分别为:(2.24±0.64)、(2.05±0.29)、(1.52±0.28)、(0.93±0.19)个,与正常组(0.82±0.18)比较,除第12组外均增多(P＜0.01);黏膜下层血管口径分别为:(4.52±1.51)、(4.05±1.23)、(3.75±1.11)、(2.03±0.86)μm,除第12组外均增大(P＜0.01);脾指数分别为:(4.21±0.93)、(4.06±0.68)、(3.84 4±0.71)、(3.31±0.69)除12号组外也较正常增加(P＜0.01).结论 缩窄门静脉主干可成功制成大鼠门静脉高压症模型;其最佳缩窄口径应该是:大鼠体重200 g左右时用8号针头(直径0.8mm),大鼠体重300 g左右时用9号针头(直径0.9 mm).     

Predictive value of local and core laboratory echocardiographic assessment of cardiac function in patients with chronic stable angina: The ACTION study.

AIMS: To evaluate the relationship between echocardiographic cardiac function and outcome in patients with stable symptomatic angina. METHODS: Baseline echo left ventricular ejection fraction and volume data measured in a central laboratory was available for 7016 patients (92% of the total) participating in the ACTION trial (A Coronary disease Trial Investigating Outcome with Nifedipine GITS). Ejection fraction was also measured by investigators. Evaluation of the different echocardiographic variables was based on adjusted hazard ratios comparing the unfavourable limit of the 90% range of the variable concerned to the favourable limit. RESULTS: The centrally measured ejection fraction was the most powerful predictor of all-cause death (adjusted hazard ratio=2.5), myocardial infarction, any stroke or transient ischaemic attack and overt heart failure (adjusted hazard ratio=4.5). The addition of either end systolic volume or end diastolic volume to ejection fraction did not materially affect the power of prediction. Compared to the central ejection fraction measurement, the investigator-measured ejection fraction was a less powerful predictor for all outcomes considered. CONCLUSION: Routine echocardiography carefully analysed by standardised methods provides useful prognostic information in patients with stable angina, including for total mortality.     

脾动脉起始部真性动脉瘤的手术治疗(附7例报告)

目的：探讨脾动脉起始部真性动脉瘤的手术治疗方法，总结临床治疗经验。方法：回顾性总结1996年1月至2006年8月收治的7例病人，经彩色超声、CT和血管造影检查证实脾动脉起始部真性动脉瘤；均在全身麻醉下行动脉瘤切除，5例行脾动脉血管重建，2例同时行脾脏切除。结果：手术后10—14d治愈出院。随访1—9年，1例人工血管转流术后2年死于急性心肌梗塞，余6例均健康生活。无复发。结论：脾动脉起始部真性动脉瘤切除、脾动脉重建是—种较好的治疗方案。     

肝动脉解剖变异在肝癌介入治疗中的临床意义

目的探讨肝动脉解剖变异的DSA表现及其在肝癌肝动脉栓塞化疗（TACE）中的临床意义。方法回顾分析223例肝癌患者的DSA资料，对存在解剖变异的肝动脉进一步分析其起源、走行、分布及对TACE的影响，统计每组变异肝动脉的发生率。结果223例中，32例存在肝动脉解剖变异，占14．34％，共40支变异肝动脉，其中变异肝总动脉2支（5％，2／40），变异肝固有动脉1支（2．5％，I／40），变异肝右动脉24支（60％，24／40），变异肝左动脉13支（32．5，13／40），有30支变异肝动脉直接参与肿瘤供血，其中24支行TACE治疗，6支行肝动脉化疗（TAI）。结论正确认识肝动脉的解剖变异特征对提高TACE的成功率、降低手术风险有极其重要的意义。     

脑源性神经生长因子和神经干细胞联合移植对大脑中动脉阻塞大鼠神经功能恢复的作用

目的 将脑源性神经生长因子(BDNF)和神经干细胞(NSCs)单独及联合移植应用于大脑中动脉阻塞(MCAo)模型大鼠,观察BDNF和NSCs移植对大鼠缺血性脑卒中神经功能恢复的作用及BDNF对内源性和外源性NSCs增殖、迁移及分化的影响.方法 体外分离、培养新生大鼠海马NSCs,BrdU标记.实验动物随机分为A组(MCAo组);B组(MCAo+BDNF组);C组(MCAo+NSCs组);D组(MCAo+BDNF+NSCs组),每组16只,移植后进行神经功能损害评分(NSS),用免疫组织化学行BrdU、nestin、BrdU/NSE检测,分析结果.结果 移植后的2、4周神经功能评分分别为:A组5.3±0.5、5.3±0.5;B组4.0±0.8、3.8±0.5;C组3.5±0.6、3.5±0.6;D组2.0 ±0.8、1.8±1.0,D组显著好于其他3组(P<0.05),B组与c组显著好于A组(P<0.05).nes.tin阳性细胞数:A组1.24±1.13,B组2.59±1.44(P<0.05),BrdU阳性细胞数:A组0.52±0.68,B组1.65±1.10(P<0.05).BrdU阳性细胞数:C组6.08±1.52,D组10.26±1.96(P<0.05),BrdU/NSE双阳性细胞数:C组1.74±1.04,D组3.58±1.20(P<0.05).结论 BDNF和NSCs移植单独及联合应用对MCAo大鼠的神经功能恢复均有作用,两者联合具有协同作用.BDNF对内源性NSCs的激活、增殖有促进作用,对外源性NSCs的增殖、迁移及分化有促进作用.     

Treatment strategy for synchronous metastases of colorectal cancer: is hepatic resection after an observation interval appropriate?

Background In cases of synchronous colorectal hepatic metastases, the primary colorectal cancer strongly influences on the metastases. Our treatment policy has been to conduct hepatic resection for the metastases at an interval of 3 months after colorectal resection. We examined the appropriateness of interval hepatic resection for synchronous hepatic metastasis. Materials and methods The subjects were 164 patients who underwent resection of hepatic metastasis of colorectal cancer (synchronous, 70 patients; metachronous, 94 patients). Background factors for hepatic metastasis and postoperative results were compared for synchronous and metachronous cases. Results The cumulative survival rate for 164 patients at 3, 5, and 10 years postoperatively was 71.9%, 51.8%, and 36.6%, and the post-resection recurrence rate in remnant livers was 26.8%. Interval resection for synchronous hepatic metastases was conducted in 49 cases after a mean interval of 131 days. No difference was seen in postoperative outcome between synchronous and metachronous cases. Conclusion The outcome was similarly favorable in cases of synchronous hepatic metastasis and in cases of metachronous metastasis. Delaying resection allows accurate understanding of the number and location of hepatic metastases, and is beneficial in determining candidates for surgery and in selecting surgical procedure.     

慢性乙型肝炎患者病毒e系统状态与肝纤维化关系的研究

目的:观察慢性乙型肝炎患者乙型肝炎病毒e系统状态和复制指标在肝纤维化发生过程中的变化及其与血清纤维化标志的关系,探讨它们在肝纤维化发生过程中的作用及其可能的临床意义.方法:188例慢性乙型肝炎患者根据肝纤维化程度分为S0～S4期等5组,分别用定量PCR及放免法检测患者血清中HBV-DNA及肝纤维化标志透明质酸、Ⅳ型胶原、Ⅲ型前胶原和层粘连蛋白的含量;HBeAg和抗-HBe采用酶联免疫吸附法(ELISA)检测,并观察其在不同肝病理纤维化分期时的变化.结果:随着肝纤维化程度加重,血清HBV-DNA含量逐渐升高,从S1期开始显著增加(P＜0.01);而HBeAg阳性率逐渐降低,S3、S4期较S0显著减少(P＜0.05和P＜0.01);抗-HBe阳性率呈相反的变化趋势,在S3和S4期的阳性率明显高于S0期(P＜0.05和P＜0.01).血清HBV-DNA( )HBeAg( )组血清纤维化标志最低,HBV-DNA(-)抗-HBe( )组最高,两者差异有显著性(P＜0.01).结论:HBV复制和e系统状态的改变与肝纤维化程度密切相关,肝内病毒复制标志与血清纤维化标志联合检测,对于判断肝纤维化程度和指导抗病毒治疗有重要的价值.