Establishment and identification of the human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM

In this study, we aim to establish the human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM which can be grow and passaged steadily in 1 μg/ml concentration of adriamycin in appropriate medium. The human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM was established using the method of exposure to medium with adriamycin alternated between high and low concentration with gradually increasing concentration. Furthermore, QBC939 and QBC939/ADM were both treated with adriamycin, mitomycin and vindesine, and then detected by MTT assay, respectively. Growth cycle and intra-cellular concentrations of ADM within cells of each group were determined by flow cytometry. Expression levels of P-glycoprotein were detected by Western bolt and real-time PCR. Results showed that, compared with QBC939, the inhibitive rates of adriamycin, mitomycin and vindesine to QBC939/ADM were lower. Content of ADM in the QBC939/ADM was lower. Western bolt and real time PCR showed that P-glycoprotein in the QBC939/ADM group was over expressed. Therefore, QBC939/ADM was establish and identified as the multi-drug-resistant cell line, which can grow and be passaged steadily in 1 μg/ml concentration adriamycin in appropriate medium. And the multi-drug-resistant character of QBC939/ADM was indicated to be related to the over expression of P-glycoprotein induced by chemotherapeutic drugs.     

我国余甘子新害虫锡兰玻壳蚧的形态及生物学特性

【目的】锡兰玻壳蚧Drepanococcus chiton是我国近年来新纪录的一种蜡蚧科(Coccidae)有害昆虫，对农业生产及生态环境造成巨大威胁，在云南滇西首次发现并记述其危害余甘子Phyllanthusemblica。研究明确其空间分布及相关形态学和生物学特性对该害虫的监测与防控意义重大。【方法】采用标准地调查于2019年5-9月对云南保山3块余甘子栽培林内的锡兰玻壳蚧种群发生密度进行抽样调查，应用聚集度指标法分析空间分布型；2020年1-12月分别以鲜嫩无虫余甘子枝条和嫁接有锡兰玻壳蚧雌成虫的余甘子植株作为寄主进行室内培养和试验地饲养，观察锡兰玻壳蚧的生殖方式、胚后发育、形态特征、发育历期及生活史。【结果】锡兰玻壳蚧在滇西余甘子栽培林内呈聚集分布，为害率为32%~56%，虫口密度为10.95~94.26头/株。该蚧虫一年发生2代，以受精雌成虫在寄主枝条上越冬，产卵期为1-2月、6-7月和10-11月，1-2龄若虫发生期为1-3月和6-9月，雄成虫羽化及雌雄交尾高峰期分别在3月中下旬和7月下旬-8月中旬。生殖方式为两性生殖，胚后发育系卵生类型；卵椭圆形、橘黄色，单产堆积于母体外被蜡壳保护，平均发育历期8-13 d；1龄若虫体扁平光滑，橘黄色，行动活泼随风扩散，12-18 d蜕皮；2龄若虫体背稍拱，分泌出齿状蜡质突起，7-10 d分化雌雄；3龄雌若虫体背大幅拱起，具透明薄蜡壳，虫体黄绿色或棕褐色，平均历期13-18 d；雌成虫平均历期最长(50-62 d)，产卵前腹部向背面挤压，单雌产卵量847.03±13.72粒，卵孵化率为97.59%；雄性历经预蛹、蛹和成虫3个发育阶段，预蛹期5~7 d，蛹期7~10 d，雄成虫前翅发达，后翅退化，外生殖突发达，生命历期最短仅1~3 d，林间很难见到。【结论】滇西余甘子林中的锡兰玻壳蚧种群呈聚集分布，雌成虫种群具一定产卵繁殖能力，薄弱期为1龄若虫分散期和2龄若虫刺吸发育期(分别为1-3月和6-9月)，建议这些时期对该虫开展综合防治。     

Dynamic Modeling of a Man–Land System in Response to Environmental Catastrophe

A compatible relationship between “man” and “land” is the essence of regional sustainable development. In this article VENSIM PLE is used to set up a system dynamics (SD) model of the regional man–land system of Xiangfan, Hubei Province, China. The model is used to simulate dynamic behaviors of society and economy in the face of environmental catastrophe, in order to explore dynamic response among subsystems in man–land systems, and to research mechanisms of regional sustainable development. The model is also used to forecast impacts from the south-to-north water transfer project in China on the environment, society, and economy within a time span of 50 years. The model is further used to verify the effects of suggested regulatory policies. The simulation results show counterintuitive outcomes. The most serious impact on Xiangfan caused by the water transfer project would be on economic growth, not on environment quality. However, if a series of regulatory policies were carried out in advance of the water project's construction, the trends of concern could be effectively eased. SD provides tools for precisely understanding complex man–land systems, which always seem counterintuitive. However, there still exist limitations and disadvantages in the model that must be overcome. It is believed that the next logical step in deriving better dynamic models of man–land systems is to integrate SD with other advanced algorithms or technologies.     

Tamoxifen reverses the multi-drug-resistance of an established human cholangiocarcinoma cell line in combined chemotherapeutics

Our previous study established the human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM. In this study, we investigate further the ability of tamoxifen (TAM) to reverse drug-resistance to chemotherapeutics using QBC939/ADM cells. Cell growth inhibition was determined by the MTT assay, while cell cycle progression, apoptosis and the intra-cellular concentration of adriamycin (ADM) were all determined by flow cytometry. P-glycoprotein (P-gp) protein and mRNA expression was determined by Western blotting and real-time PCR. Growth inhibition and apoptosis induced by ADM, mitomycin (MMC), or vindesine (VDS) were enhanced after pre-treatment with 5 or 10 μM TAM, while only VDS increased cell numbers in the G₂/M phase. The intra-cellular concentration of ADM rose after pre-treatment with 10 μM TAM, but not 5 μM TAM. Furthermore, real-time PCR and western blot analysis revealed down-regulation of P-gp expression in QBC939/ADM cells after TAM pre-treatment. The enhanced effects of TAM on growth inhibition, apoptosis, and intra-cellular concentration and the down-regulation of P-gp expression were blocked by an anti-P-gp antibody. TAM (10 μM) may reverse the multi-drug-resistance (MDR) of QBC939/ADM and enhance the chemotherapeutic effects on cholangiocarcinoma, by competitively inhibiting over-expressed P-gp.     

MiR-127-3p targeting CISD1 regulates autophagy in hypoxic–ischemic cortex

Neonatal hypoxic–ischemic (HI) injury derived from asphyxia during perinatal period, is a serious complication of neonatal asphyxia and the main cause of neonatal acute death and chronic neurological injury. Aberrant autophagy occurs in many nervous system diseases, but its role and underlying mechanism in HI injury is largely unknown. Here, we successfully constructed a newborn rat model of HI brain injury, and the knockout-miR-127-3p (KO-miR-127-3p) rats were structured by using CRISPR/Cas9. Subsequently, the in vitro functional experiments, in vivo zea-longa scores, as well as bioinformatics analyses and biological experiments were applied. The expression of autophagy-related proteins, including ATG12, P62, Beclin-1, LC3II in HI cortex with miR-127-3p knockout was significantly decreased, and autophagic vacuoles were disappeared. Moreover, miR-127-3p has a specific regulatory effect on CISD1 expression, another crucial molecule in autophagy process. Accordingly, the overexpression of CISD1 effectively inhibited the autophagic cell death and physiological dysfunction in the brain of HI injury, whereas si-CISD1 reversed the neuroprotective effects of KO-miR-127-3p. Our findings explained the underlying mechanism for HI injury, and miR-127-3p targeting CISD1 signal could be supposed as a new treatment strategy to prevent and treat HI injury.Subject terms: Autophagy, Molecular biology     

探究式课程设计在医学生临床决策能力培养中的探索

临床决策能力是一个医生的必备能力之一，是医学教育中必要且重要的培养目标之一。目前对医学生临床决策能力的培养没有形成完备的体系，一般安排在高年级阶段进行，为时较晚。如何把临床决策能力的培养融入低年级基础课中，以尽早开展能力训练值得探索研究。本研究以医学遗传学课程为例，结合探究式教学方式的特点，模拟临床决策思维过程的各细分模块，优化了课程设计，探索在低年级课程中融入临床决策能力的培养。结果提示：探究式课程设计能充分调动学生主动性，增加学生分析、思考的投入；在教师引导下，促使学生依循临床决策思维的过程得到有效地训练。最后，整理分析了探究式课程设计及实践的思路，总结了设计原则和要点，反思了教学中的经验得失。     

High prevalence of irritable bowel syndrome with poor sleep quality in children and adolescents in Shanghai

Sleep and Biological Rhythms - This study explored the prevalence of irritable bowel syndrome (IBS) and investigated the relation-ship between IBS and disturbed sleep in school-age children in...