BLOC⁃3介导Rab32线粒体定位改变对肝癌细胞生长的作用研究

目的 观察溶酶体相关细胞器生物发生复合体⁃3 （biogenesis of lysosome⁃related organelles complex⁃3,BLOC⁃3）亚基Hps1和Hps4对肝癌细胞中Rab32线粒体定位的影响及在肝癌细胞生长中的作用。 方法 通过公共数据库GenDoma,String和InBio Discover分析Hps1和Hps4与Rab32的相互作用情况。体外培养人肝癌细胞系SNU⁃739和Hep⁃3B,利用脂质体转染方法分别转染Hps1和Hps4相关的siRNAs和质粒,采用免疫荧光和Western blot观察Hps1和Hps4对Rab32线粒体定位的影响,细胞划痕、克隆形成、EdU、MTS及Transwell侵袭实验检测Hps1和Hps4调控Rab32线粒体定位后肝癌细胞迁移、增殖和侵袭的变化情况。通过公共数据库UALCAN中的数据集CPTAC分析Rab32蛋白在肝癌和正常肝脏组织中的表达差异。 结果 数据库分析结果显示,Hps1和Hps4均可以与Rab32相互作用;与正常肝脏组织相比,Rab32蛋白在肝癌组织中的表达显著降低（P<0.001）。在肝癌细胞系SNU⁃739中干涉Hps1或Hps4或同时干涉Hps1和Hps4后,Rab32线粒体定位均减少（均P<0.01）,细胞线粒体Rab32蛋白表达均降低（均P<0.001）,细胞增殖、迁移和侵袭能力增强（均P<0.05）;在肝癌细胞系Hep⁃3B中过表达Hps1和Hps4后,Rab32线粒体定位增多（P<0.01）,细胞线粒体Rab32蛋白表达增加（P<0.001）,细胞增殖、迁移和侵袭能力均受抑制（均P<0.01）,而单独过表达Hps1或Hps4时无显著抑制作用（均P>0.05）。结论 BLOC⁃3亚基Hps1和Hps4均可与Rab32相互作用并增加Rab32线粒体定位,进而抑制肝癌细胞生长。     

Primmorph extracts and mesohyls of marine sponges inhibit proliferation and migration of hepatocellular carcinoma cells in vitro

Cancer recurrence and severe side effects of currently being used chemotherapeutic agents reduce their clinical efficacy. Thus, there is a constant need to develop alternative anticancer drugs. Sustainable supply is an important challenge facing marine-based drug discovery. Primmorph, a 3D cell culture system, could provide a sustainable source to produce metabolites for anticancer drugs from marine sponges. In the present work, the anticancer activity of primmorph extracts and mesohyls of Negombata magnifica, Hemimycle arabica, Crella spinulata, and Stylissa carteri sponges was evaluated. Antiproliferative activity was studied in terms of cytotoxicity, colony formation, cell cycle, and apoptosis. Migration was assessed by migration assay and matrix metalloproteinase activity. The expression of proliferation and migration-related genes was analyzed using real time PCR. Migration and proliferation activities of HepG2 cells were inhibited by treatment with primmorph extracts and mesohyls of N. magnifica, H. arabica, and C. spinulata. The mesohyl of S. carteri did not show any anticancer activity although the primmorph extract led to cell cycle arrest. Among the selected sponge species, the primmorph extract of C. spinulata was the most promising anticancer agent regarding antiproliferative and antimigratory activities. In addition, primmorph extracts have the advantage of working under well-defined and controlled conditions, which allows the easy application as a bioreactor.     

miR-125a-5p对卵巢癌细胞的增殖迁移和侵袭能力的影响

目的探讨miR-125a-5p对卵巢癌细胞增殖、迁移和侵袭的影响及其作用的靶基因。方法购入正常人卵巢上皮细胞HOSEpi C,卵巢癌细胞SKOV3、A2780、ES2和HO8910各1株,通过实时定量聚合酶链式反应(RT-PCR)检测人卵巢上皮细胞HOSEpi C以及卵巢癌细胞SKOV3、A2780、ES2和HO8910中miR-125a-5p的表达水平。利用生物信息学方法预测miR-125a-5p的靶基因,构建预测靶基因3’端非编码区(3’-UTR)的野生型(WT)和突变型(mut)荧光素酶报告基因质粒,通过双荧光素酶报告基因法验证miR-125a-5p与预测靶基因的靶向作用。利用慢病毒感染的方法建立稳定过表达miR-125a-5p的SKOV3细胞,使用荧光定量PCR和Western blot检测靶基因的表达水平。利用细胞计数试剂盒-8(CCK-8)分析过表达miR-125a-5p的SKOV3细胞增殖能力的变化,通过Transwell迁移和侵袭试验分析过表达miR-125a-5p的SKOV3细胞迁移和侵袭能力的变化。结果正常人卵巢上皮细胞HOSEpi C中miR-125a-5p的表达水平高于卵巢癌细胞SKOV3、A2780、ES2和HO8910中miR-125a-5p的表达水平,其中SKOV3中miR-125a-5p的表达水平最低,差异均有统计学意义(P <0. 05),选择SKOV3进行后续实验。生物信息学分析显示,miR-125a-5p能够靶向结合Rab3D的3’-UTR;双荧光素酶报告基因分析证实miR-125a-5p能够靶向作用于Rab3D的3’-UTR。筛选稳定表达miR-125a-5p和si-Rab3D及相应对照的SKOV3细胞,分别命名为SKOV3/miR-125a-5p组,SKOV3/si-Rab3D组,SKOV3/miR-NC组和SKOV3/si-NC组。在SKOV3/miR-125a-5p组中,Rab3D的表达水平低于SKOV3组和SKOV3/miR-NC组,差异均有统计学意义(P <0. 05)。细胞增殖能力检测结果显示,与SKOV3组和SKOV3/miR-NC组相比,SKOV3/miR-125a-5p组在72小时和96小时细胞增殖能力降低,差异有统计学意义(P <0. 05)。Transwell迁移和侵袭试验结果显示,SKOV3/miR-125a-5p组分别与SKOV3组和SKOV3/miR-NC组相比,细胞迁移和侵袭能力均显著下降,差异有统计学意义(P <0. 05)。结论 miR-125a-5p能够靶向作用Rab3D,抑制卵巢癌细胞的增殖、迁移和侵袭。     

miRNA-381靶向HMGB1对IHH-4细胞生长侵袭及迁移的影响

目的探究微小RNA-381 (miR-381)调控高迁移率族蛋白B1 (HMGB1)对甲状腺乳头状瘤IHH-4细胞生长、侵袭及迁移的影响。方法购置甲状腺乳头状瘤IHH-4细胞及正常甲状腺细胞Nthy-ori-3各1株,取第3代细胞培养48 h后,采用实时定量PCR(qRT-PCR)检测miR-381和HMGB1在甲状腺乳头状瘤IHH-4细胞和正常甲状腺Nthy-ori-3细胞中的表达水平。利用Lipofectamine 2000转染miR-381 mimic后,检测IHH-4细胞中miR-381和HMGB1的表达水平。生物信息学预测miR-381和HMGB1的靶向关系,并用荧光素酶报告实验验证两者的靶向关系。每孔按2×105/m L细胞密度接种IHH-4细胞,分为IHH-4组(甲状腺乳头状瘤细胞IHH-4组)、miR-381 mimic组(miR-381 mimic转染组)、pc-HMGB1组(HMGB1过表达转染组)、mimic+pc-HMGB1组(miR-381 mimic和pc-HMGB1共同转染组),每组设3个复孔,采用CCK8法检测各组IHH-4细胞活性,Transwell及划痕实验分别检测IHH-4细胞侵袭及迁移能力,免疫印迹法检测IHH-4细胞中Ki67、基质金属蛋白酶-2(MMP-2)、MMP-9和晚期糖基化终产物受体(RAGE)的表达水平。结果与Nthy-ori-3细胞比较,IHH-4细胞中miR-381mRNA水平降低,HMGB1 mRNA水平升高,差异有统计学意义(P <0. 01)。与IHH-4组相比,miR-381 mimic组miR-381表达增加(P <0. 01)。miR-381和HMGB1存在靶向关系。与IHH-4组相比,miR-381 mimic组细胞增殖倍数、Ki67表达、侵袭细胞数、划痕闭合率、MMP-2、MMP-9和RAGE表达均降低(P均<0. 01),pc-HMGB1组上述指标均升高(P均<0. 01);与miR-381mimic组相比,mimic+pc-HMGB1组细胞增殖倍数、Ki67表达、侵袭细胞数、划痕闭合率、MMP-2、MMP-9和RAGE水平也明显上升(P均<0. 01)。结论 miR-381通过靶向下调HMGB1表达,抑制IHH-4细胞增殖、侵袭和迁移。     

Restaurer la fonction maternelle blessée par la guerre et l’exil. Fragments de thérapie transculturelle

Even if the literature about the difficulty of becoming a parent in a foreign country is now rich, there are fewer data about the impact of migration or psychological trauma on a parenting process already underway. This article aims to show the way parenthood can be harmed by traumatic events such as war and exile. Through the story of a mother who fled from sub-Saharan Africa with two of her children, we question the possibilities of putting the maternal function to work which, although impeded, can be subject to changes and reconsolidations, thanks to psychic work undertaken in the context of a transcultural setting. In this situation, the body is a privileged expression of the guilt and suffering of the family members. Considering the meaning of somatic symptoms and welcoming the singularity of each person, the transcultural group allows the family to find a narrative capacity and a way to co-construct a common narrative. The transcultural method, which recommends the use of psychoanalysis and anthropology in a non-merging and non-simultaneous way, allows therapists to rely on both singular and collective representations of the parental function. The presence of an interpreter, fulfilling the true role of cultural mediator, allows the use of the mother tongue and a privileged access to the inner intimate and emotional world of the family members in a familiar and reassuring environment. The analysis of certain parts of this therapy, bringing together in the same space a mother and her children, thus helps to clarify the various therapeutic levers that therapists resorted to in order to support the process of parenthood.     

“It&#x27;s not just about being here,but what brought you here”: A qualitative study of the role of migration experiences in shaping im/migrant women's access to healthcare

This qualitative study aimed to understand how migration experiences shape im/migrant women's needs, desire for, and expectations of healthcare in the British Columbia (BC), Canada context. Interviews with 33 im/migrant women (December 2018–January 2020) highlighted that traumatic experiences across migration increased healthcare needs; insufficient prior health system information contributed to poor experiences; and comparative healthcare experiences across places shaped future healthcare expectations. We use the BC setting to demonstrate the need to abide by global commitments to protect people during migration, train providers in trauma-informed care, develop health assessments that center migration journeys, and appropriately fund im/migrant-serving community organizations.     

2020年四川省凉山彝族自治州HIV/AIDS流动情况及相关因素分析

目的分析凉山彝族自治州(凉山州)现存活艾滋病病毒感染者/艾滋病病例(HIV/AIDS)流动情况及相关因素。方法根据我国艾滋病综合防治信息系统, 纳入凉山州2020年有随访记录的28 772例HIV/AIDS作为研究对象, 分析其流动情况, 采用多因素logistic回归模型分析流动的相关因素, 绘制HIV/AIDS流入地的疫情地图。结果 28 772例HIV/AIDS中, 2020年发生流动的占20.89%(6 010/28 772)。多因素logistic回归分析结果显示, HIV/AIDS流动的促进因素包括15～24岁年龄组(相比于0～14岁年龄组, OR=2.74, 95%CI:2.04～3.69)、彝族(相比于汉族, OR=2.44, 95%CI:2.19～2.72)、初中文化程度(相比于小学及以下文化程度, OR=1.25, 95%CI:1.14～1.38)、未婚(相比于已婚, OR=1.29, 95%CI:1.20～1.39)、职业为商业服务(相比于农民, OR=1.96, 95%CI:1.31～2.92)、抗病毒治疗时间<1年(相比于抗病毒治疗时间>3年, O...     

血管细胞粘附分子1在分化型甲状腺癌中的表达及临床意义

目的:检测血管细胞粘附分子1(VCAM1)在分化型甲状腺癌(DTC)中的表达情况,观察过表达VCAM1甲状腺细胞的侵袭和迁移能力。方法:收集医院收治的60例DTC患者临床资料,按照VCAM1蛋白表达情况将其分为阳性组(39例)和阴性组(21例)。对两组患者DTC组织及癌旁甲状腺组织分别采用实时荧光定量聚合酶链反应(PCR)法、蛋白免疫印迹和免疫组织化学法检测DTC组织及癌旁甲状腺组织VCAM1mRNA和蛋白表达情况,对比DTC组织中不同病理分级、肿瘤大小及预后的蛋白表达阳性及阴性表达情况。采用慢病毒转染法建立过表达甲状腺细胞系FTC-133-VCAM1、空载细胞系FTC-133-control,采用Transwell小室及划痕试验检测并对比FTC-133、FTC-133-VCAM1及FTC-133-control细胞系的穿膜细胞数及迁移率。结果:DTC组织与癌旁甲状腺组织比较,VCAM1mRNA和蛋白相对表达量较高,差异均有统计学意义(t=43.724,t=44.423;P<0.05);DTC组织VCAM1阳性表达构成比为65.00%,高于癌旁甲状腺组织(13.33%),差异有统计学意义(χ²=33.611,P<0.05);阳性组存在淋巴结转移、复发及死亡患者占比分别为33.33%、17.95%和15.38%,显著高于阴性组,差异均有统计学意义(χ²=4.127,χ²=4.065,χ²=3.867;P<0.05);FTC-133-VCAM1细胞与FTC-133、FTC-133-control细胞相比,穿膜细胞数及迁移率较高,差异均有统计学意义(t_{穿膜细胞数}=5.107,t=5.440;t_迁移率=14.108,t=13.387;P<0.05)。结论:VCAM1在DTC中异常高表达,且与淋巴结转移及预后关系密切,推测与VCAM1高表达增强癌细胞侵袭及迁移能力有关。     

miR-143调控基质金属蛋白酶-13表达对骨肉瘤细胞迁移及侵袭的影响

目的:探究miR-143对基质金属蛋白酶-13(matrix metalloproteinase,MMP-13)表达的调节作用及其对骨肉瘤细胞迁移及侵袭能力的影响。方法:采用96孔板培养小鼠骨肉瘤细胞系143B细胞,设立空白组、阴性组、阳性组、干预组。空白组不做特殊处理,阳性组加入50 μg miR-143 mimic,阴性组加入等量mimic NC(miR-143 mimic的对照序列),干预组加入50 μg miR-143 mimic及10 μg MMP-13蛋白,继续培养3~6 h,最后吸出血清处理0.5 h。分别采用荧光定量PCR和Western-blot测定各组miR-143和MMP-13蛋白表达,采用Transwell、划痕实验测定细胞的侵袭与迁移能力。结果:阳性组及干预组MMP-13蛋白表达量明显低于空白组,阳性组低于干预组(P<0.05);空白组、阴性组、阳性组及干预组每个视野的侵袭细胞数均值分别为(1 000.01±44.77)、(959.25±46.32)、(245.04±4.33)、(634.06±33.78)个;阳性组及干预组划痕愈合率均明显低于空白组,阳性组低于干预组(P<0.05)。结论:MMP-13是miR-143作用的一个靶点,可以通过抑制MMP-13表达降低骨肉瘤细胞的迁移及侵袭能力。