反相高效液相色谱法测定犬血浆中左氧氟沙星浓度及其犬体内药代动力学

目的 :建立测定犬体内左氧氟沙星血药浓度的反相高效液相色谱法 ,并测定其犬体内药代动力学。方法 :以环丙沙星为内标 ,采用 10 %高氯酸为沉淀剂处理犬血浆样品 ,色谱柱为DIKMADiamon silTM C18,5m ,15 0mm× 4 .6mm ,流动相为甲醇 PBS 三乙胺 异丙醇 (33 6 7 0 .14 0 .4 ,用磷酸调pH为 3.0 ) ,流速 1.2ml·min-1,柱温 2 0℃ ,荧光检测(λex=2 95nm ,λem=4 40nm)。结果 :血药浓度线性范围 0 .2 5～ 5 0mg·L-1(r =0 .9998) ,最低检测限浓度为 0 .0 2mg·L-1(S N >3) ,绝对回收率为 83.0 %～85 .6 % (n =5 ) ,方法回收率为 92 .10 %～ 10 6 .77%(n =5 ) ,日内和日间RSD分别为 1.3%～ 4 .0 %和1.8%～ 8.2 % (n =5 )。结论 :本方法简便、灵敏、精密度高 ,重现性好 ,适用于左氧氟沙星犬体内药代动力学研究及生物等效性研究     

甲磺酸加替沙星片治疗急性细菌性感染的细菌学疗效

目的评价甲磺酸加替沙星片治疗急性细菌性感染疾病的细菌学疗效. 方法以盐酸左氧氟沙星片为对照药,采用多中心随机盲法阳性对照平行试验设计方法,评价甲磺酸加替沙星片治疗急性细菌性感染的细菌学疗效. 结果甲磺酸加替沙星片组和盐酸左氧氟沙星片组细菌分离阳性率分别为77.42%和80.99%;两组细菌清除率分别为 97.92%和97.96%;对不同致病菌感染有效率分别为83.33%～100%和80%～100%;以上结果两组差异无统计学意义;纸片药敏试验结果表明加替沙星、左氧氟沙星、氧氟沙星、环丙沙星和司帕沙星的细菌敏感率分别为93.81%、93.30%、91.24%、84.54%和87.11%;从MIC90结果看,加替沙星对多数致病菌的抗菌活性略强于左氧氟沙星. 结论甲磺酸加替沙星片具有良好的细菌学疗效.     

玻璃体腔注射康柏西普对不同OCT分型糖尿病黄斑水肿的疗效观察

目的：观察玻璃体腔注射康柏西普对不同OCT分型糖尿病黄斑水肿(DME)的疗效差异。

方法：DME患者96例96眼根据OCT形态特征将DME分为弥漫性黄斑水肿(DRT,35眼)、囊性黄斑水肿(CME,33眼)、浆液性视网膜脱离(SRD,28眼),所有患者行玻璃体腔注射0.5mg(0.05mL)康柏西普治疗。治疗后1、3、6mo,比较三组BCVA(LogMAR)、黄斑中心凹视网膜厚度(CFT)、注射次数及视力提高眼数的差异。

结果：随访6mo,三组患眼BCVA呈明显下降趋势(F_时间=205.880,P_时间<0.01),三组间比较有差异(F_组间=3.472,P_组间=0.042),其中DRT组治疗后BCVA改善最佳; 三组患眼CFT呈明显降低趋势(F_时间=392.994,P_时间<0.01),三组间比较有差异(F_组间=5.046,P_组间=0.012),DRT组和CME组的CFT降低程度优于SRD组。随访6mo DRT组的注射次数最少,视力提高眼数比例最高。

结论：玻璃体腔注射康柏西普可显著改善不同OCT分型DME患眼的视力,降低CFT,其中DRT疗效最佳,注射次数最少。     

眼附属器淋巴瘤的临床病理分析

目的：探讨眼附属器淋巴瘤(OAL)的发病情况、临床表现及病理分型,以提高临床诊治水平。

方法：选取2005-08/2013-07在华西医院眼科确诊的OAL患者147例,收集患者的一般情况、临床表现、临床检查(影像学、病理学)结果、治疗及预后情况等临床资料进行回顾性研究。

结果：本研究纳入OAL患者147例,其中男91例(61.9%),女56例(38.1%); 年龄3.5～87岁; 病程20d～10a; 发病部位以眼眶最多见(106/147); 最常见的临床表现为眼部包块、眼睑肿胀、眼球突出; 病理分型以黏膜相关淋巴组织边缘区B细胞淋巴瘤最多见(112/147),且其预后最好,而弥漫大B细胞淋巴瘤、结外NK/T细胞淋巴瘤,鼻型预后最差。

结论：OAL病理类型复杂,发病部位和临床表现多样,免疫组织化学染色是OAL鉴别诊断的重要依据,其预后与病理类型密切相关。     

应用非接触性眼表综合分析仪观察T2DM和/或高血压患者的眼表特征

目的：使用非接触性眼表综合分析仪分析2型糖尿病(T2DM)和/或高血压患者眼表特征。

方法：选取2017-01/2018-12在我院就诊的T2DM患者、高血压患者、T2DM合并高血压患者、无T2DM也无高血压患者各40例40眼,分析比较四组患者OSDI评分、泪河高度、眼红指数、角膜荧光素钠染色评分、非侵入性泪膜破裂时间(首次泪膜破裂时间和平均泪膜破裂时间)检测结果。

结果：T2DM患者眼部不适的主观感受更为明显,T2DM和/高血压患者更容易出现眼部充血的体征。与无T2DM也无高血压的患者相比,T2DM患者的泪河高度降低,平均泪膜破裂时间明显缩短(均P<0.05),但四组患者的首次泪膜破裂时间、角膜荧光素钠染色评分均无明显差异(P>0.05)。

结论：非接触性眼表综合分析仪能够无侵入地观察眼表特征,提供相对客观的量化检查结果。T2DM患者眼部稳定性较无T2DM也无高血压的患者下降,稳态更容易被打破。     

氯化锂通过TGFBI促进角膜基质成纤维细胞增殖和自噬

目的：研究TGFBI和微管相关蛋白轻链3(LC3)在角膜营养不良患者中的表达,及氯化锂(LiCl)通过TGFBI对角膜基质成纤维细胞增殖能力的影响。

方法：用免疫组化和Western-blot方法检测角膜营养不良及正常角膜组织中TGFBI和LC3的表达。实验构建了TGFBI过表达载体并转染角膜基质成纤维细胞,分别以5、10、20、40mmol/L LiCl作用于突变型TGFBI转染的角膜基质成纤维细胞,检测不同时间(0、1、6、12h)后,TGFBI与LC3蛋白表达变化,并用CCK-8法检测细胞增殖活性。

结果：TGFBI和LC3在角膜营养不良患者角膜组织中显著高表达。TGFBI过表达抑制角膜基质成纤维细胞增殖活性(P<0.05)。LiCl抑制突变型TGFBI转染的角膜基质成纤维细胞中TGFBI和LC3蛋白表达,并增强其细胞增殖活性(P<0.05)。

结论：LiCl可以促进角膜基质成纤维细胞增殖活性和自噬,其作用机制与下调TGFBI和LC3的表达有关。     

Constructing Large Scale Cohort for Clinical Study on Heart Failure with Electronic Health Record in Regional Healthcare Platform: Challenges and Strategies in Data Reuse

Regional healthcare platforms collect clinical data from hospitals in specific areas for the purpose of healthcare management. It is a common requirement to reuse the data for clinical research. However, we have to face challenges like the inconsistence of terminology in electronic health records (EHR) and the complexities in data quality and data formats in regional healthcare platform. In this paper, we propose methodology and process on constructing large scale cohorts which forms the basis of causality and comparative effectiveness relationship in epidemiology. We firstly constructed a Chinese terminology knowledge graph to deal with the diversity of vocabularies on regional platform. Secondly, we built special disease case repositories (i.e., heart failure repository) that utilize the graph to search the related patients and to normalize the data. Based on the requirements of the clinical research which aimed to explore the effectiveness of taking statin on 180-days readmission in patients with heart failure, we built a large-scale retrospective cohort with 29647 cases of heart failure patients from the heart failure repository. After the propensity score matching, the study group (n=6346) and the control group (n=6346) with parallel clinical characteristics were acquired. Logistic regression analysis showed that taking statins had a negative correlation with 180-days readmission in heart failure patients. This paper presents the workflow and application example of big data mining based on regional EHR data.     

Overexpression of NIMA-related kinase 6 (NEK6) contributes to malignant growth and dismal prognosis in Human Breast Cancer

NIMA Related Kinase 6 (Nek6) is a protein kinase involved in various cellular processes, including cell cycle regulation, apopotosis, senescence, telomere maintainance and chemoresistance. In the present study, we investigated the role of Nek6 in breast tumorigenesis and the prognostic merit of Nek6 expression in breast cancer. Immunohistochemistry and Western blot analyses was conducted to determine the expression profile of Nek6 in 133 breast cancer specimens. Nek6 was overexpressed in a majority of breast cancer specimens, compared with the adjacent non-tumorous tissues. Furthermore, we revealed that high expression of Nek6 was associated with histologic grade, tumor size and TNM stage in breast cancer. Liner regression analysis showed a significant association between the levels of Nek6 and Ki-67. Cox regression analysis indicated that Nek6 expression was an independent prognostic predictor for breast cancer. Moreover, intracellular level of Nek6 was remarkably increased following the release from serum starvation in MCF-7 cells. EdU incorporation assay indicated that depletion of Nek6 remarkably impaired the proliferation of MCF-7 cells. Finally, spheroid formation assay revealed that interference of Nek6 led to diminished oncospheroid-forming capacity of breast cancer cells. In conclusion, our results imply that Nek6 plays a facilitating role in breast cancer cell proliferation and may serve as a promising therapeutic target for breast cancer.