刚地弓形虫磷酸甘油酸变位酶2基因片段克隆、表达及抗原性分析

目的克隆、表达刚地弓形虫(Toxoplasma gondii)磷酸甘油酸变位酶2(TgPGAM2)基因片段,并分析其抗原性。方法提取弓形虫RH株速殖子总RNA,逆转录合成cDNA。PCR扩增TgPGAM2基因。扩增产物经双酶切后连接入pET30a(+)载体,重组质粒转化大肠埃希菌(E.coli)DH5α,阳性菌落经PCR和双酶切鉴定,并测序。将测序正确的重组质粒pET30a(+)-TgPGAM2转化至E.coli BL21并加入异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)结合考马斯亮蓝染色检测表达产物。以兔抗弓形虫血清为一抗,蛋白质印迹(Western blotting)分析重组蛋白的抗原性。结果PCR扩增产物约为750 bp。菌落PCR、双酶切和测序结果显示,重组质粒pET30a(+)-TgPGAM2构建成功。SDS-PAGE结果显示,经IPTG诱导获得相对分子质量(Mr)约30 000的可溶性重组蛋白。Western blotting分析证实其能被兔抗弓形虫血清识别。结论刚地弓形虫RH株TgPGAM2基因片段可在原核表达系统中表达,且该可溶性重组蛋白具有抗原性。     

脂联素在糖调节受损患者冠状动脉病变风险中的意义

目的 探讨脂联素在不同糖调节受损者冠脉病变发生及严重程度中的临床意义.方法 疑似冠心病行冠脉造影者210例,分为正常血糖组(NGT) 42例,空腹血糖调节受损组(IFG) 36例,糖耐量受损组(IGT)92例(其中IGT1组44例2 h 血糖＜10 mmol/L;IGT2组48例2h血糖≥10mmol/L),IFG+IGT组40例.检测体重指数、血压、血脂、胰岛素、脂联素、C反应蛋白,并进行Gensini评分.结果 IGT、IFG+IGT组冠心病患病率及Gensini评分显著高于IFG及NGT(P＜0.05);脂联素在IGT、IFG+IGT两组中显著低于IFG、NGT组(P＜0.05),C反应蛋白在IGT、IFG+IGT两组显著升高(P＜0.05);IGT2组较IGT1组Gensini显著升高、脂联素显著下降(P＜0.05);Gensini与脂联素负相关,与C反应蛋白、HOMA-IR正相关(p＜0.05);多因素逐步回归分析显示脂联素和HOMA-IR是冠脉病变严重程度独立影响因素.结论 低脂联素血症可更敏感地预测IGR患者的冠脉病变程度,尤其在IGT组中,如餐后血糖控制在10 mmol/L以下可能会带来更大的心血管获益.     

伤椎置钉并椎体内植骨内固定术在胸腰椎骨折治疗中的应用

目的观察伤椎置钉并椎体内植骨内固定术在胸腰椎骨折治疗中的应用效果。方法至少一侧椎弓根完整的胸腰椎骨折26例,予伤椎撬拨复位置钉、椎体内植骨、椎弓根内固定,据胸腰段正侧位片比较术前、术后即刻、术后6个月的椎体高度及Cobb’s角,并评估脊髓神经功能和融合情况。结果术后随访7～28个月。患者术后即刻伤椎椎体前缘高度占正常高度百分比及Cobb’s角优于术前(P均<0.05);伤椎外形恢复满意;按ASIA脊髓神经功能分级,伴神经损伤者15例中,A级1例无恢复,余均改善1～3级(P均<0.05),内固定无失效,植骨皆融合。结论伤椎置钉并椎体内植骨内固定术治疗胸腰椎骨折可重建椎体结构,增强脊柱稳定,近期疗效较好。     

老年糖尿病合并冠心病患者的窦性心率震荡分析

目的：探讨老年糖尿病（DM）合并冠心病（CHD）患者窦性心率震荡（HRT）变化的意义。方法：选择伴有室性早搏的老年DM合并CHD患者46例（DM＋CHD组）、老年DM非CHD患者52例（DM组）和老年健康人56例（健康对照组）,计算各组震荡起始（TO）及震荡斜率（TS）,并进行比较。结果：与健康对照组比较,DM组和DM＋CHD组TO值明显升高[（0.43±2.11）比（1.64±2.79）比（3.72±3.32）],TS值明显下降[（11.33±1.29）比（9.26±2.05）比（6.96±2.62）],P均〈0.05;多支病变较单支病变有TO上升,TS下降的趋势,但差异无显著性（P〉0.05）。结论：窦性心率震荡可作为老年糖尿病合并冠心病患者自主神经系统功能的完整性和稳定性及预后的指标之一。     

血小板减少的系统性红斑狼疮患者骨髓巨核细胞量与临床特点的关系

目的 分析血小板减少的系统性红斑狼疮(SLE)患者的临床、实验室特点,探讨骨髓巨核细胞数量对患者临床特征的影响.方法 回顾性分析北京大学人民医院住院102例血小板减少的SLE患者,以无血小板减少的140例SLE患者为对照,比较2组在器官受累、疾病活动度、骨髓象、免疫学特征及疗效方面的差别.统计学方法采用x2检验和t检验.结果 75例患者血小板减少发生在SLE确诊前后1年之间.血小板减少发生在SLE确诊前的患者中,血小板下降程度更为显著.有血小板减少的SLE患者脏器受累比例较高,主要包括神经系统和肺,但2组在总体SLE疾病活动指数(SLEDAI)评分方面的差异无统计学意义.根据骨髓巨核细胞数量分组,16例患者巨核细胞减少,无巨核细胞减少的患者34例.巨核细胞减少组的患者无论在疾病活动度、脏器受损等临床特点方面与无巨核细胞减少组差异无统计学意义.血小板减少的近期治疗效果也与骨髓巨核细胞数量无相关性.结论 合并血小板减少的SLE患者容易合并其他重要脏器受损.     

Proteomic analysis of glutathione S-transferase isoforms in mouse liver mitochondria

AIM: To survey glutathione (GSH) S-transferase (GST) isoforms in mitochondria and to reveal the isoforms’ biological significance in diabetic mice.METHODS: The presence of GSTs in mouse liver mitochondria was systematically screened by two proteomic approaches, namely, GSH affinity chromatography/two dimensional electrophoresis (2DE/MALDI TOF/TOF MS) and SDS-PAGE/LC ESI MS/MS. The proteomic results were further confirmed by Western blotting using monoclonal antibodies against GSTs. To evaluate the liver mitochondrial GSTs quantitatively, calibration curves were generated by the loading amounts of individual recombinant GST protein vs the relative intensities elicited from the Western blotting. An extensive comparison of the liver mitochondrial GSTs was conducted between normal and db/db diabetic mice. Student’s t test was adopted for the estimation of regression and significant difference.RESULTS: Using GSH affinity/2DE/MALDI TOF/TOF MS, three GSTs, namely, alpha3, mu1 and pi1, were identified; whereas five GSTs, alpha3, mu1, pi1, kappa1 and zeta1, were detected in mouse liver mitochondria using SDS-PAGE/LC ESI MS/MS, of these GSTs, GST kappa1 was reported as a specific mitochondrial GST. The R² values of regression ranged between values of about 0.86 and 0.98, which were acceptable for the quantification. Based on the measurement of the GST abundances in liver mitochondria of normal and diabetic mice, the four GSTs, alpha3, kappa1, mu1 and zeta1, were found to be almost comparable between the two sets of animals, whereas, lower GST pi1 was detected in the diabetic mice compared with normal ones, the signal of Western blotting in control and db/db diabetic mice liver mitochondria is 134.61 ± 53.84 vs 99.74 ± 46.2, with P < 0.05.CONCLUSION: Our results indicate that GSTs exist widely in mitochondria and its abundances of mitochondrial GSTs might be tissue-dependent and disease-related.     

抗焦虑对胃癌根治术患者七氟烷用量及术后恢复的影响

目的:研究劳拉西泮联合咪达唑仑对择期胃癌根治术患者抗焦虑作用的有效性,及该方法对术中七氟烷用量和术后疼痛的影响.方法:观察组手术前1d服劳拉西泮,术前40min静注咪达唑仑,术中BIS监测控制麻醉深度,观察两组患者术前Spielberge状态—特质焦虑问卷(STAI)评分,术中七氟烷用量,麻醉恢复时间及术后2、6、24h疼痛VAS评分.结果:观察组状态-特质焦虑问卷评分低于对照组(S-AI:53.60±4.45vs62.70±3.98,P<0.05;T-AI:54.78±3.97vs65.65±4.21,P<0.05),七氟烷的使用量少于对照组(34.70±0.46vs36.20±0.44,P<0.05).两组术后苏醒及拔管时间无统计学差异.两组患者术后6、24hVAS评分,观察组低于对照组(6hVAS评分:3.45±1.60vs4.89±1.91,P<0.05;24hVAS评分:3.51±1.76vs5.17±1.71,P<0.05).结论:劳拉西泮联合咪达唑可以减轻术前患者的焦虑状态,减少术中七氟烷的使用,有利于减轻患者术后疼痛.     

Neuro-cognitive impairment in breast cancer patients: pharmacological considerations

Post-chemotherapy cognitive impairment has been an issue of concern in cancer survivors. While most reviews are focused on patient-related factors, it is proposed that drug-related factors may also be determinants. The objective of this review is to study the relationship between the types and dose intensities of chemotherapy regimens on cognitive impairment in breast cancer patients through a systematic literature search. Eighteen prospective studies were selected. The types, dose intensities and durations of chemotherapy regimens received by subjects were compared against prevalence results obtained in individual studies. It was observed that the duration of impairment varied across different generations of chemotherapy regimens. Concurrent administration of multiple cytotoxic agents can lead to a synergistic decline on cognition. Current clinical evidence is insufficient to evaluate the relationship between the types, dose intensities of chemotherapy regimens and cognitive impairment. More investigation is needed to examine the role of pharmacological factors in chemotherapy-associated cognitive changes.