Cdc42、VEGF及CD44v6在乳腺浸润性导管癌中的表达及其意义

目的探讨乳腺浸润性导管癌(invasive ductal carcinoma,IDCa)中细胞分裂周期蛋白42(cell divisioncycle 42,Cdc42)、细胞粘附分子(Homing cell adhesion molecule variant6,CD44v6)及血管内皮生长因子(Vascuoarendothelial growth factor,VEGF)的表达及其与临床病理参数的关系。方法采用免疫组织化学SP法检测Cdc42、VEGF、CD44v6在69例乳腺IDCa中的表达。结果在乳腺IDCa中Cdc42与VEGF的表达呈正相关(P<0.05),Cdc42与CD44v6的表达没有相关性(P>0.05),在乳腺IDCa中VEGF与CD44v6的表达呈正相关(P<0.05)。Cdc42、VEGF、CD44v6在69例乳腺IDCa组织的阳性表达率分别为86.96%、88.41%、78.26%。Cdc42在有或无淋巴结转移组、不同病理分级组表达差异有统计学意义(P<0.05);VEGF在不同肿瘤直径组、不同临床分期组表达差异有统计学意义(P<0.05);CD44v6在有或无淋巴结转移组、不同肿瘤直径组、不同临床分期组表达差异有统计学意义(P<0.05)。结论 Cdc42、VEGF、CD44v6在乳腺IDCa发生、发展中可能起协同作用,并可能成为判断预后的指标。     

胸部数字合成体层成像时的呼吸信号提取

目的利用一次模型分析在胸部数字合成体层成像（CTS）扫描过程中的扫描对象的呼吸状态。方法从CTS投影图像中提取横膈肌的运动产生呼吸信号。通过动态规划获取的横膈肌运动轨迹进行模型拟合，根据拟合数据，分离出横膈膜的基底运动曲线及呼吸信号曲线。采用物理体模数据、数字Xcat体模数据和临床患者数据进行实验，验证本文方法的性能。结果物理体模数据的实验中提取出的横膈膜的运动轨迹是线性的；3组不同类型的呼吸状态下的临床数据实验中都能有效的提取出呼吸信号；Xcat仿真实验中所提取的呼吸信号与原设计的呼吸信号之间的相关系数为0.9797。结论利用一次模型能够实时地有效获取病人的呼吸运动信息。临床上医生能实时获取病人的呼吸信息，便于医生做出是否要重新扫描的判断。     

Acute cutaneous barrier disruption activates epidermal p44/42 and p38 mitogen-activated protein kinases in human and hairless guinea pig skin

Acute cutaneous barrier disruption of the skin elicits various homeostatic repair responses in the epidermis. Although several candidates for the signaling mechanisms that induce these responses have been reported, e.g. the calcium and ion concentration, peroxisome proliferator-activated receptor-alpha, and TNF-alpha signaling mediated by sphingomyelinases, the exact nature of the signals remains undertermined. Therefore, assuming that an important group of serine/threonine-signaling kinases, mitogen- and SAPK/JNK, might link the barrier disruption to the subsequent homeostatic responses, the activation of three MAPKs in hairless guinea pig or in human skin after barrier disruption was investigated. The epidermal barrier was insulated with tape stripping or organic solvents, and Western blotting, and immune complex kinase assay. In the skin of hairless guinea pigs, p44/42 MAPK and p38 MAPK, but nor SAPK/JNK, were continued to be activated for at least 180 min. The activation of p44/42 which positively correlated with the number of tape strippings, whereas K+ sucrose solution suppressed its activation. The activation of p44/42 MAPK was also induced by treatment of the skin with organic solvents. In similar fashion, p44/42 and p38 MAPKs were found to be activated in human skin after tape stripping. These results for strongly suggest that the activation of p44/42 and p38 MAPKs links the stimuli of barrier disruption to the subsequent homeostatic responses to repair the barrier defect.     

CD41-42突变型β地中海贫血重组载体pEGFP-C2-CD41-42的构建及其稳定转染HeLa细胞模型的建立

目的：构建CD41-42突变型β地中海贫血重组载体pEGFP-C2-CD41-42和HeLaCD41-42细胞模型,为β地中海贫血CD41-42突变型细胞、小鼠模型建立及进一步基因治疗提供依据。方法：在β地中海贫血CD41-42突变的纯合子患者外周血中提取DNA,PCR法扩增含有βCD41-42的β-珠蛋白基因片段。将PCR产物βCD41-42珠蛋白基因克隆到pEGFP-C2载体中,用脂质体2000将pEGFP-C2-βCD41-42质粒转染HeLa细胞,观察转染后βCD41-42在HeLa细胞中荧光表达情况。RT-PCR法检测βCD41-42在HeLa细胞中的表达。结果:重组质粒pEGFP-C2-βCD41-42转染HeLa细胞24 h后细胞发出绿色荧光,G418筛选后,有大量荧光表达。RT-PCR扩增得到227 bp的特异性条带,而稳定转染pEGFP-C2空质粒的HeLa细胞中不表达特异性条带。结论: 成功构建β地中海贫血pEGFP-C2-βCD41-42突变基因重组载体,并成功构建HeLaCD41-42细胞模型。     

冠心病患者的CD62P、CD42b检测的临床意义

对冠心病患者的CD62P、CD42b进行检测以了解血小板活化水平,可作为冠心病早期诊断的一个客观依据,同时对冠心病患者的血小板活化程度进行动态观察,可及时掌握患者的病情进展及预后判断,从而有利于临床的治疗.     

脑脊液CK、Aβ42、P-tau在阿尔茨海默病中的诊断价值

目的探究脑脊液CK、Aβ42、P-tau在阿尔茨海默病中的诊断价值。方法选取2018年5月至2020年5月本院收治的40例阿尔茨海默病患者(AD)为研究对象,另选取40例轻度认知障碍期患者(MCI),40例血管性痴呆患者(VD),40例健康体检人员为对照组。采用ELISA法测定四组脑脊液CK、Aβ42、P-tau水平;以受试者工作特征曲线(ROC曲线)评估三者诊断AD的价值。结果脑脊液CK、Aβ42、P-tau水平比较,AD组、VD组、MCI组CK及P-tau水平均显著高于健康对照组(P<0.05),AD组、VD组、MCI组Aβ42均显著低于健康对照组差异均有统计学意义(P<0.05)。AD组CK、P-tau水平显著高于VD组(P<0.05),Aβ42水平显著低于MCI组、VD组(P<0.05);VD组CK、P-tau水平显著高于MCI组(P<0.05)。ROC曲线分析结果显示,脑脊液CK、Aβ42、P-tau水平分别取3.045 U/L、155.63 ng/L、182.32 ng/mL时诊断效能最高,AUC分别为0.816、0.690、0.790(95%CI:0.709~0.923、0.572~0.808、0.678~0.902);三项联合诊断AD的AUC为0.887(95%CI:0.816~0.958),敏感度为0.950、特异度为0.725。结论脑脊液CK、Aβ42、P-tau对阿尔茨海默病有一定诊断价值,三项联合诊断价值更高。     

TNK2 promoted esophageal cancer progression via activating egfr‐akt signaling

BackgroundThis study investigated the clinical implication of TNK2 expression in esophageal cancer patients’ cancer tissue samples.MethodsThe expression of TNK2 in esophageal cancer tissues and para‐carcinoma tissue was assessed with immunohistochemistry and Western blot analysis; besides, the proteins of CDC42, EGFR, and Akt were also analyzed. Then, Kaplan‐Meier survival curves of TNK2 protein expression level were assayed with 184 esophageal cancer patients from TCGA database. Moreover, with multiple linear regression analysis, we detected the correlations of TNK2 expression associated with tumor differentiation degree and metastasis status.ResultsIt revealed that TNK2 was highly expressed in the cytoplasm of esophageal cancer tissues compared with para‐carcinoma tissue; besides, the proteins of CDC42, EGFR, and Akt were also up‐regulated in different levels of esophageal cancer tissues. However, there was no significant difference of the overall survival time of TNK2 protein expression in 184 esophageal cancer patients from TCGA database (p = 0.37). But, in the included study samples of our study, there was positive coefficience between TNK2 protein expression and differentiation degree in esophageal cancer with multiple linear regression analysis [R = 0.928, 95% confidence interval (0.085‐0.12)].ConclusionOur results indicated that TNK2 was a potential diagnostic marker and promoted esophageal cancer progression through activating EGFR‐AKT signaling.     

一例早发癫痫性脑病42型患儿的临床表型及基因变异分析

目的探讨一例早发癫痫性脑病42型患儿的基因型与表型特征。方法详细询问患儿的病史,结合其临床表型、影像学及遗传学特征进行临床诊断,并对其父母进行Sanger测序验证,明确致病变异的来源。结果患儿无意识头向一侧轻度歪斜,眼球向同侧斜视,脑电图异常放电。磁共振成像显示左额后皮层可疑异常信号,伴右侧上颌窦及筛窦炎症。全外显子组测序提示患儿携带CACNA1A基因c.5789G>A杂合变异,Sanger测序提示父母双方并未携带相同的变异,提示其为新发变异。结论先证者CACNA1A基因c.5789G>A杂合变异可能是导致其早发癫痫性脑病42型的原因。     

细胞分裂周期蛋白42高表达在雌激素诱导的乳腺癌细胞耐药性增强过程中的作用

目的 观察细胞分裂周期蛋白42(Cdc42)在雌激素作用下的变化,探讨细胞内物质运输的改变在雌激素引发的乳腺癌细胞耐药中的意义.方法 分别以100 ng/ml 17β雌二醇(E2)处理MCF-7细胞,以Cdc42的小干扰RNA(siRNA) Stealth Select RNAiTM siRNA转染MCF-7细胞.采用四甲基偶氮唑蓝法检测细胞的药物敏感性,流式细胞术检测细胞内阿霉素(ADM)的蓄积量,实时荧光定量聚合酶链反应检测细胞的Cdc42 mRNA表达,Western blot法检测细胞活化的Cdc42蛋白及总Cdc42蛋白表达量.结果 E2处理后,ADM对MCF-7细胞的半数抑制浓度(IC50)由(0.098±0.011) μg/ml增高到(0.134±0.130)μg/ml(P＜0.05),细胞内ADM的相对含量则由7.253±0.310下降为3.233±0.313(P＜0.05),而Cdc42 mRNA、活化Cdc42蛋白及总Cdc42蛋白的表达量均显著增加(P＜0.05).Cdc42 siRNA转染后,ADM对MCF-7细胞的IC50下降到(0.057±0.017)μg/ml(P＜0.05),细胞内ADM的相对含量增高为11.217±0.521(P＜0.05),而Cdc42 mRNA、活化Cdc42蛋白及总Cdc42蛋白则均有显著下降(P＜0.05).结论 雌激素可以诱导乳腺癌细胞耐药性增强,其机制可能是通过上调Cdc42基因的转录、表达和活化,加速胞内物质运输速度,使得化疗药物无法在细胞内聚集.

Abstract：

Objective To investigate the changes of Cdc42 expression under estrogen stimulation, and to explore the signaling pathway of intracellular material transportation caused by estrogen. Methods MTT was used to test the drug sensitivity of cells. Real-time PCR was used to evaluate the expression of Cdc42 mRNA. The amount of ADM accumulated in MCF-7 cells was detected by flow cytometry. The protein levels of active-Cdc42 and Total-Cdc42 were measured by Western blot. Results IC50 of ADM in MCF-7 cells was increased from (0.098±0.011)μg/ml to (0.134±0.130)μg/ml (P<0.05) after estrogen stimulation. The amount of ADM accumulated in MCF-7 cells was reduced from 7.253±0.310 to 3.233±0.313 (P<0.05). All of Cdc42 mRNA, active-Cdc42 protein and total-Cdc42 protein were increased (P<0.05). After the treatment with siRNA, the IC50 of ADM in siRNA group was decreased to (0.057±0.017)μg/ml (P<0.05) compared with that in the control group. The amount of accumulated ADM was significantly increased in the siRNA group, and all the expression levels of Cdc42 mRNA, active-Cdc42 protein and total-Cdc42 protein were decreased in the siRNA group (P<0.05). Conclusions Estrogen enhances the drug resistance in breast cancer cells. The mechanism of this effect may be via the enhancing Cdc42 expression and decreasing the accumulation of chemotherapeutic drugs in the cancer cells.