简介:AbstractObjective:To investigate the relationship between the concentration of L-carnitine in semen and sperm parameters and investigate the epigenetic profile in sperm cell after L-carnitine usage.Methods:From February 2017 to February 2018, 46 semen samples from asthenospermic males and 41 semen samples from healthy donors were acquired. Motility parameters were assessed using computer-assisted sperm analysis (CASA, n = 78) and the DNA fragmentation index (DFI) was evaluated through flow cytometry (n = 86), %DFI = % cells outside main population. Other oxidative stress markers, such as reactive oxygen species (ROS) levels (n = 86) and the mitochondria DNA copy numbers, were detected (n = 78). The concentration of L-carnitine and acetyl-L-carnitine was detected (n = 82), and methylation was analyzed (n = 30). After that, we collected 13 fresh semen samples from asthenospermic males and 23 fresh semen samples from healthy donors. These samples were used in a freeze-thaw model that was used to determine whether adding L-carnitine could change sperm progressive motility (n = 23), apoptosis index (n = 9), and methylation analysis (n = 7). In total, we have done 13 asthenospermia samples for Western blot, and except for the poor Western result, we analyzed 6 samples for H3K9ac detection, 7 samples for H3K9m3 and H3K27m3 detection, and immunofluorescence (n = 3). Finally, we had recruited 30 volunteers, and they were given oral administration of L-carnitine for 3 months and then collected semen samples at different time points for methylation analysis.Results:The concentration of acetyl-L-carnitine is negatively correlated with the %DFI value (r2 = 0.1090; P = 0.0026), and the concentration of acetyl-L-carnitine is positively correlated with sperm forward motility (r2 = 0.0543; P = 0.0458) and ROS (r2 = 0.1854; P < 0.0001), and the acetyl-L-carnitine level is negatively correlated with %DFI in asthenospermia (r2 = 0.1701; P = 0.0066), and the level of acetyl-L-carnitine in asthenospermic semen is significantly lower than the normal group (P = 0.0419). In addition, this study indicates that adding L-carnitine significantly improved sperm motility (P = 0.0325) and reduced sperm apoptosis (P = 0.0032). Importantly, Western blotting (P = 0.0429) and immunofluorescence staining results showed that the addition of L-carnitine reduced H3K9Me3 methylation level in sperm, respectively. Furthermore, semen samples from asthenospermic patients had reduced methylation levels in a specific region (16thP= 0.0003; 17thP= 0.0016) of the brain-derived neurotrophic factor (BDNF) promoter. The 16th methylation decreased with age (r2 = 0.1564; P = 0.0306), and the 17th methylation was decreased after treatment with L-carnitine for 28 days (P = 0.0341).Conclusion:L-carnitine can reduce the %DFI and also affect the methylation of the histone modification marker in sperm as a possible epigenetic regulator.
简介:【摘要】 梅钢 1780 热轧 L2 过程控制计算机系统是由上海宝信软件股份有限公司自主集成开发的, L2 过程控制计算机系统中的加热炉燃烧控制模型在计算板坯温度时需要使用到加热炉 DCS 系统中的一些数据,同时需要将模型计算的相关结果下发到 L1 加热炉 DCS 系统,这就需在 L2 过程控制计算机系统与 L1 加热炉 DCS 系统之间进行数据通讯。 本文 主要 介绍了 在上海宝信软件的 iPlature 平台下,利用其 iMultiLink 技术实现 L2 过程控制 计算机系统与 L1 加热炉 DCS 系统通讯的方法。
简介:摘要:船舶副机飞车事故并非经常发生,如果是并入电网的副机发生飞车事故,极易造成全船失电和机损事故,对船舶安全航行影响很大。针对2019年某轮“
简介:【摘要】目的 : 探讨孕产妇 α -L- 岩藻糖苷酶的变化及意义。 方法 : 选取我院 2019 年 60 例患者为研究对象,均分成两组,为孕产妇组和非孕产妇组。入院时为每个患者测量 α -L- 岩藻糖苷酶含量, 24 周后再为每位患者测量一次,对比两组患者在入院前后 α -L- 岩藻糖苷酶含量的变化。 结果 : 在 24 周后,孕产妇组的 α -L- 岩藻糖苷酶含量 变化 过多,而非孕产妇的 α -L- 岩藻糖苷酶含量 变化 较少。 结论 : 孕妇在做检查时,应先检查 α -L- 岩藻糖苷酶含量变化,再决定是否患有肝病。
简介:摘要目的探讨原发性肝癌(肝癌)患者外周血循环肿瘤细胞(CTC)的程序性死亡-配体1(PD-L1)表达及其与肝癌患者临床病理特征的关系。方法回顾性分析2017年1月至2019年12月中山大学孙逸仙纪念医院收治的30例肝癌患者临床资料。患者均签署知情同意书,符合医学伦理学规定。其中男27例,女3例;年龄29~77岁,中位年龄51岁。多重RNA原位分析检测患者外周血中CTC分型及其PD-L1表达水平,分析PD-L1表达与临床病理特征之间的相关性。相关性分析采用χ2检验或Fisher确切概率法。结果共检出CTC 275个,其中上皮型82个,间质型51个,混合型142个。混合型PD-L1平均表达水平为0.89,明显高于上皮型的0.55及间质型的0.49。混合型CTC为主型肝癌患者数量及PD-L1表达水平亦明显高于其他分型患者。PD-L1阳性的CTC分型与患者肝硬化及肝被膜侵犯相关(χ2=9.707,7.153;P<0.05)。肝癌患者平均PD-L1表达水平仅与患者饮酒史相关(P=0.042)。结论肝癌患者CTC以混合型为主,其PD-L1表达水平高于其他分型患者。PD-L1阳性的CTC分型与肝硬化及肝被膜侵犯相关,而PD-L1表达水平仅与患者饮酒史相关。
简介:摘要目的探讨计算机辅助成角双L型截骨术矫正颧骨肥大的临床效果。方法回顾性分析2014年6月至2019年4月唐都医院烧伤整形科进行颧骨截骨内推术的35例女性患者的临床资料。根据患者意愿选择手术方式,其中计算机辅助成角双L型截骨26例(计算机辅助组),年龄(28.4 ± 6.6)岁;常规手术9例(常规手术组),年龄(27.3 ± 4.8)岁。计算机辅助组患者行手术导板指引下成角双L型截骨,预弯钛板辅助颧骨截骨块就位,钛板、钛钉固定。常规手术组患者行L型截骨,内推后以钛板、钛钉固定。记录手术时间,观察术后并发症(骨不连、双侧不对称)发生情况,术后3个月调查患者满意度。计算机辅助组比较手术模拟与术后3个月CT的差异。对2组的手术时间进行独立样本t检验;对患者满意度和不对称发生率进行卡方检验。以P<0.05为差异有统计学意义。结果计算机辅助组手术时间(85.1 ± 17.8) min,常规手术组手术时间(62.2 ± 11.7) min,2组比较差异有统计学意义(t=3.53,P=0.020);2组患者术后早期均未发生明显截骨块移位,也未出现明显软组织下垂,常规手术组有1例患者右侧颧骨发生骨不连,左侧颧骨骨块部分吸收。计算机辅助组不对称发生率为3.8%(1/26,不要求手术纠正),常规手术组为33.3%(3/9,其中1例要求手术纠正),2组比较差异有统计学意义(χ2=6.179,P=0.046);计算机辅助组患者满意度为100%(26/26),常规手术组为78%(7/9),2组比较差异有统计学意义(χ2=7.929,P=0.019);计算机辅助组患者术后CT与手术模拟CT图像显示,颧骨块的位置偏差为(0.21±0.19) mm。结论采用计算机辅助成角双L型截骨术治疗颧骨肥大,可以实现颧骨的精确截骨和截骨块的精确就位,可提高患者满意度,并可降低骨不连和双侧不对称等并发症的发生率。
简介:摘要目的构建肿瘤相关抗原表皮生长因子受体特异性嵌合抗原受体(EGFR-CAR)和程序性死亡受体-配体1(PD-L1)抗体双修饰慢病毒载体表达系统。方法人PD-L1-Fc蛋白免疫BALB/c小鼠,经细胞融合、亚克隆筛选高分泌PD-L1特异性抗体的稳定杂交瘤,酶联免疫吸附试验(ELISA)和Western blot检测抗体特异性,流式细胞术(FACS)鉴定对PD-1配受体封阻性能,Fortebio测定抗体亲和力,抗体全长测序,经保留鼠源CRD1、CRD2和CRD3人源化改造后构建单链抗体(single-chain variable fragment,scFv);人EGFR单克隆抗体杂交瘤系,经5′RACE技术扩增其轻链和重链可变区(VL和VH)基因,构建scFv,克隆至真核载体pcDNA3.1表达鉴定。基因合成EGFR-CAR(引入CD137协同信号胞内功能域)与PD-L1-scFv借助2A序列连接,克隆入慢病毒pLVX-EF1a-IRES-ZsGreen1表达载体,使用Lenti-X Packaging Single Shots (VSV-G)共同转染293T细胞,获得包装病毒,感染293V细胞,FACS测定CAR膜表达,ELISA检测CAR感染293V细胞培养上清中PD-L1-scFv表达情况,转染激活人外周血T细胞,验证CAR膜表达。结果获得PD-L1抗体11E3,具备高度配受体封阻性能,经人源化改造后,亲和力稳定(2.67×10-10 mol/L),EGFR-scFv获得有效表达。进一步构建了EGFR-CAR和PD-L1双修饰慢病毒分泌型CAR(CTC0537-1)及膜表达型CAR(CTC0537-2),其病毒感染293V细胞阳性率为10%。CTZ0431-1感染293V细胞后,细胞膜表面表达EGFR-scFv,检测培养上清存在PD-L1-ScFv;CTZ0431-2感染293V细胞后,细胞膜表面EGFR-scFv和PD-L1-scFv有效表达,双表达病毒感染活化T细胞的CAR表达率为39.3%。结论成功构建了EGFR-CAR和PD-L1-scFv双表达慢病毒载体,EGFR-CAR中度结合亲和力,此为EGFR靶向和PD-L1抗体双修饰CAR-T细胞的实体瘤治疗研究提供了关键工具。