向楠治疗亚急性甲状腺炎经验

向楠教授系湖北中医药大学二级教授、主任医师、博士生导师,湖北中医名师、首届湖北省中青年知名中医、武汉中青年中医名医,培养学术继承人及硕士、博士等50余名,从事中医临床、教学、科研工作30余载,擅长应用中医药防治内分泌及代谢性疾病。余有幸跟师学习,受益颇深,现将其治疗亚急性甲状腺炎的经验总结于下,与同道共享。     

鼓膜穿刺加压灌注治疗分泌性中耳炎的临床疗效

目的探讨鼓膜穿刺加压灌注疗法治疗分泌性中耳炎的临床疗效。方法选择确诊为分泌性中耳炎患者88例为研究对象,所有患者采用中耳鼓膜穿刺鼓室灌注加压治疗。比较不同性质分泌物患者的临床疗效及治疗前后患者鼓室导抗图变化。结果胶冻状患者治疗时间最长,浆液性患者治疗时间最短,三组治疗时间比较差异有统计学意义(P<0.01)。浆液性总有效率94.0%,黏液性总有效率86.8%,胶冻状总有效率75.0%,三组比较有显著差异(P<0.05)。治疗前鼓室导图主要是B型,治疗后主要是A型,治疗前后比较差异有统计学意义(P<0.01)。结论鼓膜穿刺加压灌注治疗分泌性中耳炎具有较好的临床疗效。     

PERK信号通路介导结肠癌细胞对5-氟尿嘧啶耐药机制的研究

目的:观察PERK蛋白对结肠癌细胞药物敏感性的影响，并进一步探讨其相关作用机制。方法:结肠癌细胞系HCT116分为三组:空白对照（Control）组、下调PERK表达（si-PERK）组、阴性对照（si-NC）组；采用免疫荧光及RT-PCR验证转染效率；利用CCK-8实验检测下调PERK表达后结肠癌细胞对化疗药物5-FU的敏感性变化；Annexin V-FITC凋亡实验检测下调PERK表达对结肠癌细胞凋亡的影响；利用RT-PCR及Western Blot实验检测PERK信号通路中关键分子eIF2α、ATF4、CHOP、XIAP的mRNA及蛋白表达变化。结果:RT-PCR实验表明:与正常对照组相比，si-PERK 组mRNA的表达显著下降（P＜0.05）,免疫荧光提示转染效率达80%以上；CCK-8实验发现与si-NC组相比，5-FU对 si-PERK组细胞的半数抑制浓度（IC50）明显降低（P＜0.01）；Annexin V-FITC凋亡实验发现与si-NC组相比，si-PERK组细胞的凋亡发生率显著升高（P＜0.05）；RT-PCR及Western Blot实验发现与si-NC组相比，si-PERK组细胞中PERK信号通路下游关键分子eIF2α、ATF4、CHOP的mRNA及蛋白表达均明显降低（P＜0.05）。结论:在结肠癌细胞中，抑制PERK表达后，其可能通过下调eIF2α、ATF4、CHOP的表达促进细胞发生凋亡，从而促进细胞对化疗药物5-FU的敏感性。     

Quantile regression and empirical likelihood for the analysis of longitudinal data with monotone missing responses due to dropout,with applications to quality of life measurements from clinical trials

The analysis of quality of life (QoL) data can be challenging due to the skewness of responses and the presence of missing data. In this paper, we propose a new weighted quantile regression method for estimating the conditional quantiles of QoL data with responses missing at random. The proposed method makes use of the correlation information within the same subject from an auxiliary mean regression model to enhance the estimation efficiency and takes into account of missing data mechanism. The asymptotic properties of the proposed estimator have been studied and simulations are also conducted to evaluate the performance of the proposed estimator. The proposed method has also been applied to the analysis of the QoL data from a clinical trial on early breast cancer, which motivated this study.     

Survivin-shRNA对视网膜母细胞瘤HXO-RB44细胞自噬及凋亡的影响

目的:研究Survivin-shRNA对视网膜母细胞瘤HXO-RB44细胞自噬及凋亡的影响。方法:培养视网膜母细胞瘤HXO-RB44细胞，构建Survivin-shRNA载体。按照处理不同分为Survivin-shRNA组、GFP组和CON组。分别采用流式细胞术检测Survivin-shRNA对HXO-RB44细胞凋亡和细胞周期的影响，应用MTT法检测Survivin-shRNA对HXO-RB44细胞活性的影响，Western blot法检测Survivin-shRNA对HXO-RB44 细胞自噬相关蛋白LC3、p62 与mTOR 表达的影响。结果:流式细胞术结果表明，与Control组和GFP组相比，Survivin-shRNA组细胞凋亡率增加，差异有统计学意义（P＜0.05）；随着作用时间的延长，S期出现阻滞，48 h阻滞最强，显著高于对照组（P＜0.05）。MTT结果发现Survivin-shRNA可抑制HXO-RB44细胞增殖活性，与Control组和GFP组相比，差异有统计学意义（P＜0.05）；Western blot结果发现，与对照组相比，LC3表达量显著增加（P＜0.01）；而mTOR表达量降低（P＜0.01）。结论:Survivin-shRNA可促进视网膜母细胞瘤HXO-RB44细胞的凋亡和自噬水平，进而特异性杀伤肿瘤细胞。     

SMYD3 promotes implant metastasis of ovarian cancer via H3K4 trimethylation of integrin promoters

Detachment of cancer cells from the primary tumor and formation of spheroids in ascites is required for implantation metastasis in epithelial ovarian cancer (EOC), but the underlying mechanism of this process has not been thoroughly elucidated. To mimic this process, ovarian cancer cells were grown in 3D and 2D culture. Hey and OVCA433 spheroids exhibited decreased cell proliferation and enhanced adhesion and invasion. SMYD3 expression was elevated in ovarian carcinoma spheroids in association with increased H3K4 methylation. Depletion of SMYD3 by transient siRNA, stable shRNA knockdown and the SMYD3 inhibitor BCI-121 all decreased spheroid invasion and adhesion. Gene expression arrays revealed downregulation of integrin family members. Inhibition assays confirmed that invasion and adhesion of spheroids are mediated by ITGB6 and ITGAM. SMYD3-deficient cells regained the ability to invade and adhere after forced overexpression of SMYD3, ITGB6 and ITGAM. However, this biological ability was not restored by forced overexpression of SMYD3 in ITGB6- and/or ITGAM-deficient cancer cells. SMYD3 and H3K4me3 binding at the ITGB6 and ITGAM promoters was increased in spheroids compared to that in monolayer cells, and the binding was decreased when SMYD3 expression was inhibited, consistent with the expression changes in integrins. SMYD3 expression and integrin-mediated adhesion were also activated in an intraperitoneal xenograft model and in EOC patient spheroids. In vivo, SMYD3 knockdown inhibited tumor metastasis and reduced ascites volume in both the intraperitoneal xenograft model and a PDX model. Overall, our results suggest that the SMYD3-H3K4me3-integrin pathway plays a crucial role in ovarian cancer metastasis to the peritoneal surface.