微小RNAmiR-26a的克隆及其慢病毒表达载体的构建

目的克隆微小RNAhsa-miR-26a并构建其慢病毒表达载体。方法将PCR扩增得到的miR-26。前体序列和pIVTHM载体经双酶切后连接产生pIVTHM-miR-26a慢病毒表达载体，双酶切后测序鉴定。筛选阳性克隆。用pIVTHM-miR-26a、psPAX2和pMD2-G3质粒共转染包装细胞293FT，包装产生慢病毒，以293FT细胞绿色荧光蛋白（green fluorescent protein，GFP）的表达水平测定病毒滴度。结果经双酶切鉴定和测序证实，成功构建了miR-26a的慢病毒表达载体pIVTHM—miR-26a。倒置荧光显微镜下观察可见包装细胞293FT呈绿色荧光．并测得病毒滴度为5×10^5TU／ml。成功构建hsa—miR-26a的慢病毒表达载体，为深入研究miR-26a的生物学功能奠定了基础．     

慢性胃炎脾气虚证与脾胃湿热证的差异表达基因比较

目的：比较慢性胃炎脾气虚证与脾胃湿热证患者差异表达基因。方法:分别提取慢性胃炎脾气虚和脾胃湿热患者胃黏膜组织RNA各4例，逆转录荧光探针标记后杂交制作BiostarH-140 s基因芯片。采用荧光值ratio、生物信息学、t检验等方法分析结果，实时荧光定量PCR检测部分相关基因。结果:获得差异表达基因245条，主要为营养物质消化吸收运输、物质能量合成代谢、细胞周期增殖分化、免疫反应等相关基因；有显著意义的差异表达基因77条；实时定量PCR检测10条基因，6条与芯片结果一致。结论:慢性胃炎脾气虚和脾胃湿热2个证型基因表达存在明显差异，提示中医的临床辨证分型与基因差异表达有一定关系。     

MTHFR基因多态性对血管内皮功能指标的影响

目的：研究社区人群中MTHFR 677C→T基因突变对血管紧张素Ⅱ、前列环素和一氧化氮水平的影响。方法：整群抽取社区汉族成年居民1 146人，采用PCR－RFLP鉴定目标人群MTHFR 677位基因型；采用荧光生化技术测定血浆Hcy浓度；用镉还原法检测其血清一氧化氮水平，用放射免疫分析法检测血浆血管紧张素Ⅱ和前列环素的浓度。所有数据用SPSS 13.0 软件进行统计分析。结果：1 146例居民按MTHFR 677基因型自然分组，三组之间PGI₂、AngⅡ没有统计学差异；C/C和T/C基因型组之间血清NO水平无显著差异（P>0.05），T/T基因型组的血清NO水平较C/C和T/C基因型组都低（P＜0.01）。结论:社区人群中MTHFR 677C→T突变对血浆PGI₂、AngⅡ的影响并不明显；血清NO水平下降，可能是MTHFR 677C→T导致缺血性心脑血管病的机制之一。     

全膝关节置换后不放置引流的相关管理策略

背景:近年来,随着全膝关节置换技术提升、手术时间缩短、术中及术后止血药的应用,其总失血量不断减少,术后引流量也逐渐减少,术后快速康复理论应运而生,引流管放置也再次成为骨科医生所热议的问题,越来越多的医生认为可以常规取消放置引流,引流似乎已可有可无。目的:旨在复习既往文献,对全膝关节置换后引流放置的相关问题归纳总结,提供围术期引流管理的相关经验。方法:检索中国知网、万方、维普、中国生物医学文献数据库等中文数据库,以及PubMed、The Cochrane Library、WebofScience、EMBASE、OVID等外文数据库,以"引流,膝关节置换术"及"totalknee arthroplasty,drainage"为检索词,检索自建库始至2020年2月为止所有与主题相关的文献,总结归纳全膝关节置换后不放置引流管围术期各方面需综合考虑的要素。结果与结论:①引流管的放置问题关乎全膝关节置换手术的最终结果,与术后出血、血肿、血栓、感染等并发症的发生关系密切;②目前全膝关节置换术后不推荐常规放置引流,但不意味着不需要管理,其实际上对临床医护在围术期各方面的管理提出更高的要求,包括但不限于术前适应证选择、手术操作、止血带使用、术中及术后药物使用、物理疗法的使用等。     

人外周血T细胞活化状态对血卟啉单甲醚吸收的影响

目的：观察健康人外周血T细胞的活化状态对其吸收新型光敏剂血卟啉单甲醚(HMME)的影响。以期为进一步研究HMME-PDT对活化T细胞选择性作用提供实验依据。 方法： 以流式细胞仪检测孵育浓度和孵育时间对活化T细胞吸收HMME的影响。密度梯度法常规分离人外周血淋巴细胞，分别以多克隆刺激剂PHA和PDB+Ion刺激T细胞活化，同时设立未加刺激剂的静止T细胞作对照。在评价孵育浓度对T细胞吸收HMME的影响时，将上述处理的淋巴细胞分别与0、5、10、20、30和40 mg/L的HMME孵育1 h，经CD3抗体染色后进行T细胞HMME吸收分析。在评价T细胞活化状态对HMME吸收量与孵育时间相关性的影响时，将培养的淋巴细胞与10 mg/L的HMME分别孵育0-160 min后进行检测。 结果： T细胞的活化使其HMME吸收量的孵育浓度相关曲线以及孵育时间相关曲线较静止T细胞右移和上移。在一定孵育浓度范围内(5-20 mg/L)，活化T细胞对HMME的吸收量明显大于静止T细胞，差别具有统计学意义。然而，随着孵育浓度的进一步增加，活化T细胞与静止T细胞对HMME吸收量的差异逐渐减小。在10 mg/L HMME孵育浓度的条件下，不论活化与否，随着孵育时间的延长，T细胞内HMME的吸收量在逐渐增强。在15-60 min之间，活化T细胞HMME吸收量明显高于静止T细胞。 结论： 活化T细胞与静止T细胞HMME吸收量的差别取决于其孵育浓度和孵育时间，在一定孵育浓度和孵育时间范围内，活化T细胞HMME的吸收量明显高于静止T细胞，提示这可能有助于形成选择性删除活化T细胞的剂量关联和时间关联的HMME-PDT治疗窗口。     

从氧自由基、一氧化氮探讨四逆汤抗急性失血性休克的肝脏机制

目的:从氧自由基、一氧化氮探讨四逆汤抗急性失血性休克的肝脏机制。方法:复制急性失血性休克大鼠模型, 分为假手术对照组;单纯休克模型组;休克+生理盐水复苏组;休克+四逆注射液复苏组。四逆注射液(浓度1000g生药/L), 剂量0.1mL/200g大鼠。用生理盐水或四逆注射液治疗3h后处死动物并取组织。测定各组肝脏超氧化物歧化酶(SOD)活性、丙二醛(MDA)水平, 一氧化氮(NO)水平。采用免疫组化染色法, 观察诱导型一氧化氮合酶(iNOS)在肝细胞中的变化特点。RT-PCR观测肝细胞iNOS和内皮源性一氧化氮合酶(eNOS)基因表达的变化。结果:模型组在休克1h后SOD活性明显低于对照组(P＜0.01)、MDA水平明显高于对照组(P＜0.01)。四逆汤组复苏3h后肝组织SOD明显高于生理盐水组(P＜0.01)、MDA低于生理盐水组(P＜0.01)、NO水平明显高于生理盐水组(P＜0.01)。生理盐水组iNOS在肝细胞中染色阳性单位明显高于四逆汤组(P＜0.05)。生理盐水组促进iNOSmRNA的表达。四逆汤组eNOSmRNA的表达增强。结论:四逆汤通过清除氧自由基, 升高NO, 改善肝组织微循环, 减少诱导型iNOS表达的各种因素, 理论上减轻了NO与氧自由基生成的ONOO的细胞毒作用和血管的低反应性, 并对肝脏起到保护作用。     

四川省卫生资源配置现状综合评价——基于秩和比和密切值法

目的 综合评价四川省21市（州）卫生资源配置现状,为促进卫生健康发展和“健康中国”战略实施提供依据。方法 选取每千人口卫生技术人员数、每千人口执业（助理）医师数、每千人口注册护士数、每千人口医疗机构床位数和每千人口卫生机构数5个衡量指标,利用秩和比法（RSR）和密切值法两种方法共同评价四川省内各市（州）卫生资源配置情况,并比较两种方法结果差异。结果 21市（州）按RSR法最优分档策略分成四档,probit>6:成都、攀枝花、雅安和广元,5     

Associations of park features with park use and park-based physical activity in an urban environment in Asia: A cross-sectional study

Park use is associated with health, yet our understanding of park features related to their use is limited. Singapore's parks were audited for 30 micro-features, then geospatial analysis characterized micro-features scores for parks nearest to participants' homes. Adults (3,435) reported their park use and park-based physical activity. Using linear regression models, we found living near a park with higher micro-features scores was associated with more time in parks and park-based physical activity. Specific micro-features were associated with more park time (wildlife areas, water features, forested areas, unpaved trails (2–2.6 h/month, p < 0.05)) and with physical activity in parks (water features, forested areas, large playground, open green spaces (1.8–2.2 h/month, p < 0.05)). These findings could inform parks planning to support population-health.     

Study of electroacupuncture with different frequencies for the treatment of headache and anxiety-depression symptoms in patients with migraine: A randomized clinical trial: 不同频率电针改善偏头痛患者头痛及焦虑抑郁症状的差异性研究-随机临床试验

ObjectiveTo observe the effects of electroacupuncture (EA) with varied frequencies on headache and anxiety-depression symptoms in patients with migraine and to screen optimal frequency of EA for patients with such conditions.DesignSingle-center, randomized, controlled clinical trial was designed, and the outcome assessors and statisticians were blinded. The patients with migraine were randomized into 2 Hz EA group, 100 Hz EA group and sham-stimulation group. In each group, the changes in migraine attacks, days with headache, the scores of visual analogy scale (VAS), self-rating anxiety scale (SAS), self-rating depression scale (SDS) and migraine-specific quality-of-life questionnaire (MSQ), as well as the dosage of analgesics were observed at the baseline, during treatment, in 1, 2 and 3 months of follow-up separately.SettingPatients were enrolled in the Third Affiliated Hospital of Zhejiang Chinese Medical University, between 1^st August 2018 and 31^st July 2021.ParticipantsSixty-five migraine patients with or without aura.InterventionsIn the EA groups, the acupoints were bilateral Fengchi (GB20), Gongxue (Extra), Sizhukong (TE23), Taiyang (EX-HN5), Shuaigu (GB8), Waiguan (TE5) and Yanglingquan (GB34). Electric stimulation was exerted at GB20 and Gongxue (Extra), with 2 Hz and 100 Hz separately. In the sham-stimulation group, the shallow acupuncture was operated at the sites 1 cm lateral to GB20 and Gongxue (Extra), and on the radial side of TE5 and GB34. The output wires were cut off after attached to the acupoints. The patients in each group received the treatment 3 times weekly, once every two days, for consecutive 4 weeks. The complete intervention was composed of 12 treatments.Main outcome measuresChanges in numbers of migraine attacks at treatment phase (week 1 to week 4) from the baseline(week -4 to week 0) in patients of each groupResults(1) Changes in migraine attacks and days with headache: In the 2 Hz EA and 100 Hz EA groups, the changes for migraine attacks and days of headache were higher significantly when compared with that in the sham-stimulation group at the same time stage (P < 0.05). There was no statistical difference between two EA groups. (2) Changes of VAS score: In the 2 Hz EA and 100 Hz EA groups, the changes of VAS score were significantly higher when compared with that in the sham-stimulation group at the same time stage (P < 0.05). There was no statistical difference between two EA groups. (3) Assessment of anxiety and depression: The differences in the changes of SAS and SDS scores had no statistical significance in patients of each group at each assessment stage (P > 0.05). (4) Assessment on the quality of life: Compared with the sham-stimulation group at the same time stage, the improvement in MSQ score was increased significantly during treatment in patients of the 2 Hz EA and 100 Hz groups (P < 0.05). There was no statistical difference between two EA groups. (5) Assessment on safety and compliance: The patients of each group had sound compliance. There was no adverse events during trial, suggesting promising safety of treatment.ConclusionEA may effectively reduce the migraine attacks, and the days and intensity of headache, presenting promising safety. However, there was no significant improvement on anxiety-depression symptoms, and no significant difference between high and low frequencies of EA treatment in relieving headache and anxiety/depression symptoms in the patients with migraine.Trial registrationChiCTR1800017259     

基于复杂网络的《新针灸学》与《经络腧穴学》比较分析研究（英文）

提取《新针灸学》《经络腧穴学》中穴位名称、主治病症信息,基于复杂网络建立穴-症网络,分析两者穴位数量、相互关联程度及主治规律变化,借助拓扑学数据解释变化原因,为传统针灸知识体系的结构化、标准化研究提供具体思路和方法。共纳入《新针灸学》386穴、773种症状、形成152163个穴位配伍对,《经络腧穴学》403穴、253种症状、28755个穴位配伍对。两本教材的穴-症网络存在丰富的差异性,其所载的病症结构化程度随医学知识的更新而提升。《新针灸学》模型具有更加典型的小世界效应,或因其以病症为主要分类手段的优势体现。两本教材穴位定位与主治方面发生许多变化,学科发展、时代背景等方面是变化的主要原因。