深低温冷冻肌腱细胞活性的研究

目的研究深低温冷冻方法对肌腱细胞活性的影响,比较程序性降温和普通深低温冷冻法对腱细胞活性的影响.方法纯种SD大鼠24只（出生21 d）,随机分为3组,取双侧跟腱.新鲜肌腱对照组（A）,常规深低温冷冻组（B）,程序性降温深低温冷冻组（C）.采用相同的方法对3组肌腱细胞进行细胞培养.相差显微镜观察原代和传代后细胞的生长,绘制细胞的生长曲线,考察细胞的活性;对细胞进行成纤维细胞染色、胶原染色和对细胞进行形态观察（扫描电镜）;水解法定量分析细胞培养基中羟脯氨酸浓度的变化,检测细胞合成胶原的能力.结果原代细胞培养时A组细胞的生长速度快于B组和C组（P＜0.01）,C组细胞的生长速度快于B组（P＜0.01）,这种生长速度的差异在细胞传代后消失.细胞的形态学和组织学符合成纤维细胞形态.3组细胞培养基中羟脯氨酸浓度变化的差异无统计意义（P＞0.05）.结论经深低温冷冻处理的肌腱中仍存在具有活性的腱细胞,但数量显著少于新鲜肌腱中活细胞的数量.应用计算机控制程序性慢速降温方法处理的肌腱其活细胞的数量有所提高,但仍低于新鲜肌腱中活细胞的数量.     

VX-induced cell death involves activation of caspase-3 in cultured rat cortical neurons

Exposure of cell cultures to organophosphorous compounds such as VX can result in cell death. However, it is not clear whether VX-induced cell death is necrotic or involves programmed cell death mechanisms. Activation of caspases, a family of cysteine proteases, is often involved in cell death, and in particular, caspase-3 activation appears to be a key event in programmed cell death processes including apoptosis. In this study, we investigated VX-induced neuronal cell death, as well as the underlying mechanism in terms of its effect on caspase-3 activity. Primary cortical neuronal cultures were prepared from gestational days 17 to 19 Sprague Dawley rat fetuses. At maturation, the cells were treated with varying concentrations of VX and cell death was evaluated by lactate dehydrogenase (LDH) release. VX induced an increase in LDH release in a concentration-dependent manner. Morphological VX-induced cell death was also characterized by using nuclear staining with propidium iodide and Hoechst 33342. VX induced a concentration- and time-dependent increase in caspase-3 activation. Caspase-3 activation was also confirmed by the proteolytic cleavage of poly(ADP-ribose)polymerase (PARP), an endogenous caspase-3 substrate. These data suggested that in rat cortical neurons, VX-induced cell death via a programmed cell death pathway that involves changes in caspase-3 protease.     

Synergistic convergence of microbiota-specific systemic IgG and secretory IgA

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Lidocaine has a narrow antiarrhythmic dose range against ventricular arrhythmias induced by programmed electrical stimulation in conscious postinfarction dogs

Summary The aim of the present study was to investigate the dose-dependent antiarrhythmic efficacy of lidocaine against electrically induced tachycardias in conscious, chronically instrumented postinfarction dogs. Programmed electrical stimulation (PES) was performed in 16 dogs 8 to 21 days after a 4 h occlusion of the left anterior descending coronary artery (LAD). Infusion of saline in 8 control animals with sustained ventricular tachycardia (SVT) inducible at baseline did not affect subsequent inducibility. In the treatment group 7 of 8 animals responded with SVT and one exhibited ventricular fibrillation at baseline. After an initial bolus of 1 mg/kg lidocaine intravenously (i.v.), the drug was infused at infusion rates of 40, 80 and 120 g/kg/min (i.v.). During 80 g/kg/min lidocaine (mean plasma level 3.5 g/ml) 7 out of 8 animals displayed an antiarrhythmic response; both the lower and the higher infusion rate were associated with a smaller antiarrhythmic efficacy (3 of 8 animals responded to 40 g/kg/min and 4 of 8 to 120 g/kg/min). Licocaine did not affect ventricular refractory periods, but induced an increase in intraventricular conduction time at all infusion rates, from 66.2 ms at baseline to 67.7 ms (p<0.05), 67.7 ms (p<0.05), 70.0 ms (p<0.01) respectively.In conclusion the present study demonstrates that lidocaine is of considerable value in the management of PES-induced ventricular arrhythmias in the postinfarction phase. However there is only a small optimal therapeutic plasma level range, where lidocaine exhibits its antiarrhythmic efficacy against this type of arrhythmia; this makes a carefully titration of the drug necessary both in the experimental and in the clinical setting. Send offprint requests to K. Krejcy at the above address     

卵巢上皮性癌组织中Gab1与PD L1蛋白表达与临床病理特征及预后的相关性

【摘要】 目的 探讨卵巢上皮性癌组织中Gab1与程序性死亡配体 1（PD L1）的表达情况,并分析Gab1、PD L1蛋白与卵巢上皮性癌患者临床病理特征及预后的相关性。方法 选取我院2008年5月~2014年5月收治的86例卵巢上皮性癌患者作为研究对象,采用回顾性分析法分析其临床及随访资料,收集86份癌组织标本（观察组）及86份癌旁正常组织标本（对照组）,所有组织标本均行Gab1与PD L1蛋白检测,并根据其资料及检测结果记录所有患者年龄、体质量指数等一般临床资料及肿瘤情况、Gab1与PD L1蛋白水平及预后等。结果 观察组中Gab1及 PD L1蛋白的表达水平均显著高于对照组（P＜0.05）;随着卵巢上皮性癌患者临床分期更高、分化程度越低及出现淋巴结转移时其Gab1及 PD L1蛋白的阳性表达率也显著增高（P＜0.05）;Gab1阴性和阳性患者其3年总生存率分别为7273%和4286%,而 PD L1阴性和阳性患者其3年总生存率分别为7368%和4583%（P＜0.05）;经非条件单因素Cox 比例风险回归模型显示不同临床分期、分化程度、是否出现淋巴结转移以及Gab1和 PD L1不同表达的卵巢上皮性癌患者间其预后生存时间均存在差异（P＜0.05）;经多因素Cox 比例风险回归模型分析显示临床分期Ⅲ~Ⅳ期、低分化、出现淋巴结转移以及Gab1和 PD L1阳性均为影响卵巢上皮性癌患者预后的独立危险因素（P＜0.05）。结论 Gab1和 PD L1蛋白在卵巢上皮性癌组织中均高表达,随着病情加重其阳性表达率更高,且其阳性表达联合临床分期Ⅲ~Ⅳ期、低分化和出现淋巴结转移均为影响卵巢上皮性癌患者预后生存的独立危险因素,故可将其作为临床上评估卵巢上皮性癌患者病情进展和预后的有效指标。     

急性支气管哮喘病人外周血程序性死亡因子 -1及其配体的表达

目的探讨急性支气管哮喘病人外周血程序性死亡因子 -1(PD-1)及其配体 PD-L1的表达水平及意义。方法选择 2017年 2月至 2019年 3月无锡市第五人民医院收治的 101例急性支气管哮喘病人作为病例组,同期体检正常 40例健康者为对照组;病例组病人分为轻中度组 46例、重度组 34例、危重组 21例。采用流式细胞术法检测外周血 CD4+T淋巴细胞中 PD-1表达( CD4+PD-1)、 CD8+T淋巴细胞中 PD-1表达( CD8+PD-1)CD19+B淋巴细胞中 PD-L1(CD19+PD-L1)表达,采用单因素方差分析其与疾病严重程度的关系;采用化学发光分析仪检测呼出,气一氧化氮( FeNO)水平,酶联免疫吸附法( ELISA)检测外周血白细胞介素 4(IL-4)、白细胞介素 17(IL-17)水平。结果病例组外周血 CD4+PD-1、CD19+PD-L1表达水平及 PD-1/PD-L1值均高于对照组高［ 17.98±4.16比 5.43±1.52、3.13±0.75比 1.08±0.27、5.75±1.62比 5.03±1.11］(P<0.05);病例组 FeNO及外周血 IL-17水平高于对照组［( 29.74±6.08)μg/L比( 16.38±4.12)μg/L、(38.66±8.27)ng/L比( 24.73±5.21)ng/L］,外周血 IL-4水平低于对照组［(9.14±2.47)ng/L比( 12.86±3.65)ng/L］(P<0.05);病例组不同严重程度外周血 CD4+PD-1、CD19+PD-L1表达水平及 PD-1/PD-L1值由高至低为危重组、重度组、轻中度组(P<0.05);病例组外周血 CD4+PD-1、CD19+PD-L1与 FeNO呈正相关关系(r=0.61、0.55, P<0.05)与 IL-17呈正相关关系( r=0.51、0.50,P<0.05)与 IL-4呈负相关关系(r=.0.43、.0.38,P<0.05)。结论急性支气管哮喘病人外周血,PD-1/PD-L1(CD4+PD-1/CD19+PD-L1)表达异,常,病情越严重,其表达水平可能越高。     

慢性乙型肝炎患者不同免疫状态下外周血T细胞表面程序性死亡受体1的表达

目的 检测慢性乙型肝炎(CHB)患者不同免疫状态下外周血CD4+、CD8+T细胞表面程序性死亡受体1(PD-1)表达,探讨其与血清HBV DNA载量、ALT之间的关系.方法 收集CHB患者免疫耐受期(A组,24例)、免疫活化期(B组,64例)、HBeAg阴性(C组,23例)和11例健康对照者(D组)外周血,采用流式细胞术检测CD4+、CD8+T细胞表面PD-1表达,以实时荧光定量PCR检测血清HBV DNA,同时检测ALT.结果 B、C组CD4+、CD8+细胞表面PD-1表达显著高于A、D组(P＜0.05).C组CD8+T细胞表面PD-1表达显著高于B组(P＜0.05).相关性分析发现,不同免疫状态下CHB患者CD4+、CD8+T细胞表面PD-1表达水平与HBV DNA载量及ALT均无相关性.结论 CHB患者外周血T细胞表面PD-1表达与机体免疫状态有关.肝脏炎症损伤是影响PD-1表达的重要因素,HBV DNA载量不是主要影响原因.     

Characteristics and immune checkpoint inhibitor effects on non-smoking non-small cell lung cancer with KRAS mutation: A single center cohort (STROBE-compliant)

Kirsten rat sarcoma (KRAS) mutation (KRASm) is associated with poor prognosis in non-small cell lung cancer (NSCLC) patients. We have aimed to survey NSCLC patients harboring KRASm in Taiwan, where never-smoking lung adenocarcinoma predominates, and analyze the immune checkpoint inhibitor effect on NSCLC harboring KRASm.NSCLC patients with KRASm were enrolled and tested on programmed death-ligand 1 (PD-L1) expression using available tissue. We analyzed their clinical features, PD-L1 status, responses to ICIs, and overall survival (OS).We studied 93 patients with a median age 66.0 years, 23.7% of whom were women, and 22.6% were never-smokers. The results showed that G12C (36.6%) was the most common KRASm. In 47 patients with available tissue for PD-L1 testing, PD-L1 expression was positive in 66.0% of patients, while PD-L1 ≥50% was higher in ever-smokers (P = .038). Among 23 patients receiving ICI treatment, those with PD-L1 ≥50% experience a 45.5% response rate to ICI. There were benefits from ICI treatment on OS compared with no ICI treatment (median OS 35.6 vs 9.8 months, P = .002) for all of our patients, and for patients with PD-L1 ≥50% (median OS not-reached vs 8.4 months, P = .008). There were no differences in survival across different KRAS subtypes (P = .666).Never-smokers composed more than one-fifth of KRASm in NSCLC in Taiwan. A high PD-L1 expression was related to smoking history and responded well to ICI. ICI treatment improved the OS in NSCLC patients with KRASm, particularly those with PD-L1 ≥50%.     

正中开胸上腔静脉人工血管置换的程序化处理在胸部肿瘤外科治疗中的应用

背景与目的 上腔静脉系统受累是局部晚期胸部肿瘤较常见的一种情况,手术可能获益,但风险极高.本研究针对正中开胸入路,提出一种程序化的手术方案,旨在优化流程,使得这一类以往认为风险极高的手术能够更加安全地实施.方法 35例胸部疾患累及上腔静脉系统,经正中开胸进行人工血管置换的患者资料,分期检查明确为局部晚期.包括肺部肿瘤16例,纵隔肿瘤19例.手术方法采用从左至右的单向推进,先游离左无名静脉起始部,阻断后切断,掀起瘤体,打开心包,用人工血管桥接左无名静脉和右心耳.游离上腔静脉近心端未受侵部分后,向尾侧牵拉肿瘤,剪开右侧纵隔胸膜,结扎切断右侧乳内血管,可以充分显露右无名静脉.向左上方牵拉瘤体,于肺门上方结扎切断奇静脉,此时可以阻断右无名静脉和上腔静脉,切除中间受侵的血管,以人工血管行右无名静脉-上腔静脉桥接,完成受侵的上腔静脉系统全部替换.结果 全组病例均顺利完成手术.术后并发症包括:心律失常6例,低氧血症5例,肌无力危象1例,心脏疝1例,真菌感染2例.2例患者死亡,死亡率5.12%,分别死于心梗和肺部感染.其余33例顺利出院.平均术后住院日15 d.在10例术前出现上腔静脉综合征的患者中,除2例术中即出现人工血管内血栓形成的患者,其余8例症状均明显改善.结论 上腔静脉人工血管置换手术经程序化的处理,规范治疗的细节,在手术操作过程中可降低手术风险,本组病例手术能够安全实施的实践也支持这一点.