Objective: To detect the effect of over expressed human MIR31HG gene on the proliferation and m grat on of PC9 ce Is, and to clarify the mechan sm of oncogene MIR31 HG Methods: The ful length sequence of ong non coding RNA (LncRNA) MIR31HG was amp fed by RT PGR and cloned nto the pcDNA3 1 ( ) eukaryot c expression vector. The PG9 cells were transfected w th the pcDNA3 1 MIR31HG overexpress on vector and control vector pcDNA3 1. The construction of MIR31HG overexpress on vector was detected by enzymatic digest on identification. The stab e cell 1 nes w th overexpress on of MIR31HG (PC9 pcDNA3 1 MIR31HG∗ stab e transfection group) and control ce 1 1 nes (PC9 pcDNA3 1, empty vector group) were established by G418 drug screen ng. and the express on evel of MIR31 HG gene n stably transfected cell ine was detected by RT PGR CCK 8 method and scratch heal ng assay were used to detect the proliferation act vities and m gration ab 1 ties of PC9 eel s. Results: The agarose gel e ectrophoresis resu ts showed that the spec f c gene fragment of MIR31HG was obta ned by amplif cation successfu ly. The gene fragments of target gene and vector were produced by double enzyme digest on of MIR31HG eukaryotic expression vector. The RT PGR resu ts showed that the MIR31HG RNA expression leve in the cells n stable transfect on group was sign ficantly h gher than that in empty vector group (P< 0 05). The results of CGK 8 test showed that the pro iferat on act vities of the eels in stable transfection group were s gnif cant y higher than those in empty vector group at 24, 36 and 48 h after culture ( P<0 01). The results of scratch healing assay showed that the scratch heal ng rate of ce Is n stable transfect on group was sign ficantly h gher than that in empty vector group at 48 h after culture ( P<0 05). Conclusion: The eukaryotic overexpression vector and the PG9 eel line stably transfected w th human LncRNA MIR31 HG gene are constructed successfully, and M1R31 HG overexpression can promote the pro iferat on and m grat on of ung cancer PG9 cells.

[摘 要] 目的：探讨干扰四个半LIM结构域蛋白2（FHL2）的表达对胶质母细胞瘤U87细胞中O6-甲基鸟嘌呤DNA甲基转移酶(MGMT) 表达的影响，以及对U87细胞替莫唑胺（TMZ）耐药性的影响。方法：利用慢病毒感染技术分别将携带不同FHL2干扰序列的慢病毒（shFHL2-1#、shFHL2-4#）及其阴性对照（shN）感染U87细胞，分别命名为shFHL2-1#、shFHL2-4#和shN组；采用siRNA转染技术将siMGMT-1#、siMGMT-4#和siN转染至U87细胞，为siMGMT-1#、siMGMT-4#和siN组，qPCR和WB法验证FHL2或MGMT的敲低效果。用TMZ处理上述各组细胞（以DMSO处理组为对照），随后以CCK-8法和细胞克隆形成实验检测TMZ处理前后FHL2或MGMT敲低组细胞的增殖情况，FCM检测TMZ处理前后FHL2敲低组细胞的凋亡情况，WB法和免疫荧光法检测敲低FHL2对U87细胞中MGMT表达的影响，WB法检测TMZ处理对各组细胞中FHL2和MGMT表达水平的影响。结果：成功构建敲低FHL2或MGMT表达的U87细胞。与shN组相比，shFHL2-1#、shFHL2-4#组U87细胞的增殖能力减弱、凋亡水平升高（均P<0.01），MGMT表达水平明显降低（均P<0.01）。经TMZ处理后，与相应的DMSO处理组相比，shN组细胞中FHL2和MGMT的表达水平显著升高（均P<0.05），而细胞的增殖和凋亡均无显著变化（均P>0.05）；shFHL2-1#、shFHL2-4#组细胞中FHL2和MGMT的表达水平无显著改变（均P>0.05），但细胞增殖能力进一步显著降低、凋亡水平进一步显著升高（均P<0.01）。敲低MGMT使U87细胞增殖减慢（P<0.01），而siMGMT-1#、siMGMT-4#组细胞经TMZ处理后增殖能力进一步降低（均P<0.01）。结论：干扰FHL2表达使得U87细胞增殖减慢而凋亡加剧、MGMT表达下调，提示FHL2可能通过影响MGMT的表达调控U87细胞对TMZ的耐药性。

[摘 要] 目的：探讨脾多肽联合FOLFOX4或XELOX方案用于Ⅲ/Ⅳ期结肠癌患者术后治疗的疗效及安全性。方法：回顾性收集选择2017年1月至2020年6月期间在广东省惠州市第六人民医院接受晚期结肠癌根治术的患者160例临床资料。将患者分为脾多肽注射液、奥沙利铂、亚叶酸钙联合氟尿嘧啶组[脾多肽+FOLFOX4组（脾F组），n=80]与脾多肽注射液、奥沙利铂联合卡培他滨组[脾多肽+XELOX组（脾X组），n=80]，两组患者均于手术后8周开始进行6个疗程的治疗，治疗结束1个月后对两组患者的临床疗效、免疫水平、营养状况、生存质量及不良反应等方面进行为期2年的随访观察。结果:与脾F组相比，脾X组患者的ORR和DCR均显著提高（均P<0.05）；CD3+ T、CD4+ T、NK细胞百分率、EORTC QLQ-C30评分和PNI水平均显著升高（均P<0.05），NLR、LMR、CA125、CA199、CEA均显著下降（均P<0.05）。脾X组患者白细胞或粒细胞减少、神经毒性、口腔黏膜炎和手足综合征等毒性作用和不良反应发生率均明显下降（均P<0.05）。两组患者2年PFS和OS无明显差异（均P>0.05）。结论: 在Ⅲ/Ⅳ期结肠癌术后患者的治疗中，脾多肽注射液联合XELOX方案比联合FOLFOX4方案在改善不良反应发生率和生活质量方面具有明显优势。

Methadone maintenance treatment (MMT) greatly contributed to the successful outcomes of prevention and control on both AIDS and drug abuse in China. However, the features on drug abuse changed in the past decades, and the prevalence of new psychoactive substances abuse potentially somehow offset the achievement of MMT. This paper concised the information on research and surveys of this issue that targeting on the current situation, characteristics, related factors and relevant public health problem on new psychoactive substances abuse, among patients who have been on MMT, in China.
Adult ; China/epidemiology* ; Female ; Humans ; Male ; Methadone/therapeutic use* ; Opiate Substitution Treatment ; Prevalence ; Psychotropic Drugs/adverse effects* ; Substance Abuse Treatment Centers ; Substance-Related Disorders/epidemiology* ; Surveys and Questionnaires

[摘 要] 目的：探讨食管恶性黑色素瘤（MEM）患者的临床特征，分析以PD-1单抗为基础的免疫治疗疗效及预后的影响因素。方法：收集2011年5月至2022年6月在北京大学肿瘤医院黑色素瘤暨肉瘤内科收治的手术不可切除或者转移性MEM患者的临床资料，包括基本信息、病理资料、实验室指标、治疗方案和生存情况等。采用实体瘤疗效评价标准1.1进行疗效评估，用Kaplan-Meier曲线进行生存分析，用单因素和多因素COX回归进行预后分析。结果：共收集到有完整资料的MEM患者79例，中位年龄59.0岁。大部分患者发病时伴有进食哽噎和吞咽困难等症状，以食管下段发病最为常见，NRAS和KIT基因突变的比例较高，乳酸脱氢酶（LDH）水平升高占21.5%；其中，17例患者接受化疗为主的治疗方案，62例患者接受PD-1单抗为主的免疫治疗方案，客观有效率分别为5.9%和28.8%，疾病控制率分别为35.3%和72.9%，总生存期（OS）分别为7个月[95%CI（0，16.7）个月]和13.2个月[95%CI（9.5，16.9）个月]（P<0.05）。多因素分析显示，就诊时LDH水平、ECOG评分、是否有临床症状、是否接受PD-1单抗治疗与OS显著相关（P<0.05）。结论：MEM患者对PD-1单抗为主的免疫治疗应答较好，LDH升高、ECOG评分≥2分、就诊时有临床症状可能是预后的不良因素。

<b>Backgroundb> There is a lack of evidence on whether exposure to PM_2.5 and its constituents would affect the relationship between the dietary approaches to stop hypertension (DASH) and central obesity. <b>Objectiveb> To investigate the effect of exposure to PM_2.5 and its constituents on the correlation between the DASH dietary pattern and the prevalence of central obesity. <b>Methodsb> The data were obtained from the baseline survey of the "Xinjiang Multi-Ethnic Natural Population Cohort Construction and Health Follow-Up Study" in Urumqi. A DASH score was calculated according to intake frequency of 8 food groups, and summed from intake frequency of recommended food groups scored from 1 to 5 from low to high, and intake frequency of restricted food groups scored from 1 to 5 from high to low. A higher DASH score indicates better compliance with the DASH dietary pattern. We estimated exposure using satellite-derived PM_2.5 and a chemical transport model (GEOS-Chem) for its constituents, including organic carbon (OC), black carbon (BC), sulfate (SO₄²⁻), nitrate (NO₃⁻), ammonium (NH₄⁺), and soil dust. Central obesity was defined by waist circumference: ≥90 cm for men or ≥85 cm for women according to Criteria of weight for adults (WS/T 428—2013). A logistic regression model was used to analyze the effects of the DASH dietary pattern as well as PM_2.5 and its constituents on central obesity, and a stratified analysis was used to explore the effects of PM_2.5 and its constituents on the association between the DASH dietary pattern and central obesity. <b>Resultsb> The study included 9 565 urban residents, aged (62.30±9.42) years, with a central obesity prevalence rate of 60.75%. After adjusting for selected confounders, the DASH score Q5 group had a 17.5% lower risk of central obesity than the Q1 group (OR=0.825, 95%CI: 0.720-0.947). PM_2.5 and its constituents OC, BC, SO₄²⁻, NH₄⁺, and soil dust were positively associated with the prevalence of central obesity, but no association was observed between constituent NO₃⁻ exposure and central obesity. The stratified analysis revealed that the prevalence of central obesity was reduced in the DASH score Q5 group in participants exposed to low concentrations of PM_2.5 and its constituents NO₃⁻, NH₄⁺, and soil dust, while the protective effect of the DASH pattern on central obesity disappeared in subjects exposed to high concentrations of PM_2.5 and its constituents NO₃⁻, NH₄⁺, and soil dust. <b>Conclusionb> Exposure to PM_2.5 and its constituents NO₃⁻, NH₄⁺, and soil dust could attenuate the protective effect of the DASH pattern on central obesity.

[摘 要] 目的：利用昆虫杆状病毒表达系统（昆虫BEVS）表达糖酵解酶α-烯醇化酶（ENO1）及其3种酶活性位点缺失突变的ENO1蛋白ENO1-M1、ENO1-M2和ENO1-M3，为后续宫颈癌的代谢治疗研究奠定基础。方法：利用分子克隆技术将优化后ENO1序列插入pFastBacTM1载体，获得含有目的基因的重组质粒pFastBac-ENO1。分别缺失ENO1发挥糖酵解酶功能的3个活性位点，进行优化后将其插入pFastBacTM1载体，获得3个活性位点缺失的重组质粒pFastBac-M1、pFastBac-M2和pFastBac-M3。通过转座、转染后获得重组杆状病毒rBV-ENO1、rBV-M1、rBV-M2和rBV-M3，利用WB法对目的蛋白的表达及特异性进行检测。结果：成功扩增重组杆粒rBacmid-ENO1、rBacmid-M1、rBacmid-M2和rBacmid-M3，获得大小约2 000 bp的基因片段，与预期大小相符。昆虫BEVS可表达ENO1蛋白及其3个酶活位点缺失的重组蛋白ENO1-M1、ENO1-M2和ENO1-M3，其分子量约为52 000，与预期相符。WB法鉴定这些蛋白能与特异性标签His-tag发生反应。结论：通过昆虫BEVS成功表达目的蛋白ENO1及其酶活性位点缺失蛋白ENO1-M1、ENO1-M2和ENO1-M3，这些蛋白具有反应原性，为后续测定这些蛋白与ENO1单抗亲和力创造了条件。

[摘 要] 目的：探讨地锦草乙醇提取物（EEEH）对人结直肠癌SW480细胞生物学行为的影响及其分子机制。方法：体外培养SW480细胞，实验分为Con组、EEEH-L组、EEEH-M组、EEEH-H组、si-NC组、si-circRHOT1组、EEEH-H+pcDNA组、EEEH-H+pcDNA-circRHOT1组，分别以si-NC、si-circRHOT1、pcDNA、pcDNA-circRHOT1转染SW480细胞，采用CCK-8法、细胞克隆形成实验、Transwell实验分别检测转染后各组细胞的增殖、迁移及侵袭能力，qPCR法检测转染后各组SW480细胞circRHOT1和miR-29a-3p的表达，WB法检测各组细胞中MMP-2、MMP-9蛋白的表达。双荧光素酶报告基因实验检测circRHOT1与miR-29a-3p之间的靶向关系。结果：与Con组比较，EEEH-L组、EEEH-M组、EEEH-H组SW480细胞中MMP-2和MMP-9蛋白表达均明显降低（均P<0.05），circRHOT1的表达均降低（均P<0.05）而miR-29a-3p的表达均升高（均P<0.05）且呈剂量依赖性；细胞的存活率、细胞克隆形成数、迁移及侵袭细胞数均减少（均P<0.05）。circRHOT1可靶向负调控miR-29a-3p的表达。敲减circRHOT1可抑制SW480细胞的增殖、迁移及侵袭能力，而过表达circRHOT1则可减弱EEEH对SW480细胞增殖、迁移及侵袭的抑制作用。结论：EEEH可通过调控circRHOT1/miR-29a-3p轴而抑制结直肠癌SW480细胞的增殖、迁移及侵袭能力。

Free fatty acids (FFAs) are important substrates for mitochondrial oxidative metabolism and ATP synthesis but also cause serious stress to various tissues, contributing to the development of metabolic diseases. CD36 is a major mediator of cellular FFA uptake. Inside the cell, saturated FFAs are able to induce the production of cytosolic and mitochondrial reactive oxygen species (ROS), which can be prevented by co-exposure to unsaturated FFAs. There are close connections between oxidative stress and organellar Ca²⁺ homeostasis. Highly oxidative conditions induced by palmitate trigger aberrant endoplasmic reticulum (ER) Ca²⁺ release and thereby deplete ER Ca²⁺ stores. The resulting ER Ca²⁺ deficiency impairs chaperones of the protein folding machinery, leading to the accumulation of misfolded proteins. This ER stress may further aggravate oxidative stress by augmenting ER ROS production. Secondary to ER Ca²⁺ release, cytosolic and mitochondrial matrix Ca²⁺ concentrations can also be altered. In addition, plasmalemmal ion channels operated by ER Ca²⁺ depletion mediate persistent Ca²⁺ influx, further impairing cytosolic and mitochondrial Ca²⁺ homeostasis. Mitochondrial Ca²⁺ overload causes superoxide production and functional impairment, culminating in apoptosis. This vicious cycle of lipotoxicity occurs in multiple tissues, resulting in β-cell failure and insulin resistance in target tissues, and further aggravates diabetic complications.
Adenosine Triphosphate ; Apoptosis ; Calcium* ; Cytosol ; Diabetes Complications ; Endoplasmic Reticulum ; Fatty Acids, Nonesterified ; Homeostasis ; Insulin Resistance ; Ion Channels ; Metabolic Diseases ; Metabolism ; Oxidative Stress* ; Protein Folding ; Reactive Oxygen Species ; Superoxides

[摘 要] 目的：通过分析非小细胞肺癌顺铂敏感株及耐药株的基因芯片表达数据，筛选差异基因及关键通路，构建蛋白相互作用网络，探讨关键集群功能。方法:从GEO数据库获得基因芯片表达数据，利用GEO2R工具筛选差异基因，通过STRING数据库和Cytoscape软件构建蛋白相互作用网络，经DAVID富集得到相关特征基因与信号通路信息。结果：通过芯片分析共获得481个差异表达基因，相比于敏感细胞株，顺铂获得性耐药细胞株中有418个上调基因和63个下调基因。差异基因功能主要富集在piRNA代谢、DNA甲基化修饰、细胞有丝分裂及细胞周期进程等信号通路。蛋白复合物预测得到主要功能集群6个，分别与细胞趋化性、细胞角化性、piRNA代谢过程、细胞因子受体相互作用、细胞因子分泌调节及染色质沉默相关生物进程相关。结论:本研究利用生物信息学方法，发现顺铂耐药细胞株特征基因及信号通路，其中SAA1、KRT5、TDRD9、BCL2A1、CSF1R和HIST1H1A等显著上调基因及其功能集团可能是非小细胞肺癌顺铂耐药的潜在分子机制，为临床精准治疗提供新的理论依据。