微小RNA-494-3p靶向配子生成素结合蛋白2促进结直肠癌细胞侵袭、迁移及上皮间质转化的实验研究

目的 研究微小RNA-494-3p(miR-494-3p)影响结直肠癌细胞迁移、侵袭及上皮间质转化(EMT)的分子机制.方法 于2019年1—12月体外培养购自美国ATCC的结直肠上皮细胞NCM460和结直肠癌细胞系T84、LS1034和HCT116,实时定量聚合酶链式反应(RT-qPCR)检测细胞中miR-494-3...     

Association between age at arrival,duration of migration,and overweight/obesity in Chinese rural-to-urban migrants: the Yi migrant study

Background:Urbanization in China is rapidly proceeding, but rural-to-urban migration and its association with overweight and obesity is not well studied. This study aimed to explore the age at arrival, duration of migration, and the corresponding association with overweight/obesity in Yi migrants in China.Methods:A cross-sectional study was conducted in rural and urban areas in 2015 in Sichuan province, China. Demographic characteristics, lifestyle factors, and anthropometry were collected. General linear regression models were used to assess the effect of duration of migration (1–10, 11–20, 21–30, and >30 years) on body mass index (BMI). Multi-variable logistic regression was used to examine the association between duration of migration and overweight/obesity (BMI ≥ 25 kg/m²).Results:A total of 3056 Yi people (1894 Yi farmers and 1162 Yi migrants) aged 20 to 80 years were enrolled. After adjusting for age, sex, and other potential confounders, Yi migrants had 1.71 kg/m² (95% confidence interval [CI]: 1.36–2.06) higher BMI and a 2.13-fold (95% CI: 1.71–2.65) higher risk of overweight/obesity than Yi farmers. In Yi migrants, stratified by age at arrival, no significant association between duration of migration and overweight/obesity was observed in those who were 0 to 20 years old at arrival. In comparison, in migrants >20 years old at arrival, compared with the reference group (1–10 years), long-term migration (>30 years) was found to be associated with overweight/obesity after adjustment (odds ratio: 1.85, 95% CI: 1.04–3.29).Conclusions:Yi migrants were observed to have greater risk of overweight/obesity than Yi farmers. In Yi migrants, the risk of overweight/obesity increased according to the duration of migration, especially in those who were older upon their arrival.     

江西省市售不锈钢餐具中24种元素迁移量调查

目的 了解江西省市售不锈钢餐具中24种元素迁移含量,为江西省食品安全风险监测工作提供数据支持。方法 从江西省11个地市的超市、农贸市场、网店采集65份不锈钢餐具,采用电感耦合等离子体质谱法分析测定。结果 65份餐具均有多种元素迁移检出,样品总检出率为100.0%,24种元素迁移总量范围为3.35×10 - 2～1.76 mg/kg。铅、铬、镍元素的迁移检出率分别为23.1%、100.0%、87.7%,最大检出值分别为3.39×10 - 4、0.137、1.35×10 - 2 mg/kg,均未超过限量标准。结论 江西省市售不锈钢餐具中元素迁移普遍存在,建议制定相关标准,加强监管力度。     

The chrondroitin sulfate proteoglycan (CSPG4) regulates human trophoblast function

Introduction

Trophoblast growth and invasion of the uterine endometrium are critical events during placentation and are tightly regulated by locally produced factors. Abnormal placentation can result in early miscarriage or preeclampsia and intrauterine growth restriction, leading to impaired fetal and/or maternal health. Chondroitin sulfate proteoglycan 4 (CSPG4) is involved in cancer cell migration and invasion, processes which are critical during placentation but unlike in cancer, trophoblast invasion is highly regulated. CSPG4 expression and function in trophoblast is unknown. We determined CSPG4 expression in human first trimester placenta and implantation sites, and investigated whether CSPG4 influenced proliferation, migration and invasion of a human extravillous trophoblast (EVT) cell line (HTR8/SVneo cells) as a model for extravillous trophoblast (EVT).

Methods and results

Immunoreactive CSPG4 localized to EVT cells in the trophoblast shell, subpopulations of interstitial EVT cells within the decidua and cytotrophoblast cells in placental villi. In HTR8/SVneo cells, siRNA knockdown of CSPG4 stimulated proliferation and decreased migration/invasion. In primary first trimester placental villi explants two cytokines, interleukin 11 (IL11) and leukemia inhibitory factor (LIF) with known roles in trophoblast function, stimulated CSPG4 mRNA expression and immunoreactive protein in the cyotrophoblast.

Discussion and conclusion

This is the first demonstration of the production and function of CSPG4 in human placentation. These data suggest that locally produced CSPG4 stimulates human EVT migration and invasion and suggests that IL11 and LIF regulate villous cytotrophoblast differentiation towards the invasive phenotype at least in part via CSPG4.     

胰蛋白酶抑制剂对人肝癌细胞SK－HEP-1迁移和侵袭的影响

目的 观察尿胰蛋白酶抑制剂(UTI)对人肝癌细胞株SK-HEP-1迁移和侵袭等肿瘤生物学特性的影响.方法 噻唑蓝(MTT)比色法检测UTI对SK-HEP-1细胞增殖的影响;Transwell法检测不同浓度UTI对细胞迁移和侵袭能力的作用;逆转录-聚合酶链反应(RT-PCR)和Western blot检测不同浓度UTI作用后,尿激酶型纤溶酶原激活物(UPA)的mRNA和蛋白表达.结果 UTI组对SK-HEP-1细胞的增殖无影响(P＞0.05);与对照组比较,不同浓度的UTI组均能抑制人肝癌细胞SK-HEP-1的体外迁移和侵袭能力,且呈剂量效应关系,差异有统计学意义(P＜0.05);不同浓度的UTI组中SK-HEP-1细胞UPA的mRNA和蛋白表达均明显下降,且呈剂量依赖性,与对照组比较差异有统计学意义(P＜0.05).结论 UTI能抑制人肝癌细胞SK-HEP-1迁移与侵袭,其机制可能与UTI抑制SK-HEP-1细胞UPA表达有关.     

Proximal loading of the femur leads to low subsidence rates: first clinical results of the CR-stem

Introduction

A new femoral stem was developed with a design that leads to compression of cancellous bone in the calcar region which results in proximal loading. The cross-sectional design of the implant provides rotational stability.

Materials and methods

In the first clinical investigation ten patients underwent uncemented total hip arthroplasty between January 1999 and May 1999 using the CR-stem^® (Implantcast GmbH, Buxtehude, Germany). Results were investigated using the Harris-hip-score (HHS) and antero-posterior and lateral radiographs. Migration was evaluated with the EBRA-FCA-method with a follow-up of 7 years.

Results

We demonstrated a mean subsidence rate of 2.23 ± 1.13 mm 7 years after implantation thus providing basic data for extensive testing in a clinical environment.

Discussion

As small subsidence rates are regarded as predictor for superior long-term results in uncemented total hip arthroplasty according to the literature, the CR-stem shows promise for excellent long-term results.

     

miR-424对非小细胞肺癌细胞株A549迁移及侵袭的影响

目的探讨miR-424对非小细胞肺癌细胞株A549迁移及侵袭的影响。方法 RT-PCR法检测肺癌细胞NCI-H460、NCI-H1975、NCI-H446、A549、NCI-H1299、NCI-H157及人胚肺成纤维细胞MRC-5中miR-424表达,脂质体LipofectamineTM2000将miR-424 inhibitor和miR-424 NC转入A549细胞中,48 h后,RT-PCR法检测miR-424表达,CCK-8法检测细胞活力,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力,western blot检测基质金属蛋白酶2(MMP2)、MMP9、转化生长因子-β1(TGF-β1)及p-Smad3的表达。结果 miR-424在肺癌细胞NCIH460、NCI-H1975、NCI-H446、A549、NCI-H1299、NCI-H157中的[(1.78±0.13),(1.69±0.10),(1.89±0.18),(2.88±0.27),(2.52±0.20),(2.49±0.23)]表达量显著高于miR-424在人胚肺成纤维细胞MRC-5中的(0.58±0.05)表达量(P0.01)。与miR-424 NC组比较,miR-424 inhibitor组miR-424表达量显著降低(P0.01),细胞活力下降(P0.01),细胞迁移及侵袭能力降低(P0.01),MMP2,MMP9,TGF-β1及p-Smad3表达量均显著下调(P0.01)。结论 miR-424表达量下调后能通过抑制TGF-β1/Smad3信号通路进而抑制A549细胞的迁移及侵袭。     

miRNA-186靶向Twist-1促胃癌侵袭、迁移的机制研究

目的检测miRNA-186在胃癌中的表达情况,观察miRNA-186表达改变对人胃癌细胞株侵袭、迁移能力的影响并分析可能的分子机制。方法RT-PCR法检测40例胃癌患者胃癌及配对的正常胃黏膜组织中miRNA-186的表达水平,同时检测miRNA-186在胃癌细胞株和正常胃黏膜上皮细胞中的表达水平。在胃癌细胞中分别转染miRNA-186模拟物（mimic）、抑制物（inhibitor）以上调、下调miRNA-186的表达,应用Transwell试验观察其对胃癌细胞侵袭、迁移能力的影响。应用生物信息学软件预测miRNA-186可能的靶基因,并经双荧光素酶试验及Westernblot试验验证靶基因,分析其促转移的分子机制。结果胃癌组织相较于正常胃黏膜组织、胃癌细胞株相较于正常胃黏膜上皮细胞株的miRNA-186表达水平均降低（均P＜0.05）。在胃癌细胞中,分别上调、下调miRNA-186的表达水平,能够抑制、促进胃癌细胞的侵袭、迁移能力。miRNA-186与Twist-1之间存在反向调控的关系,Twist-1是miRNA-186的靶基因。同时发现,下调miRNA-186的表达时,Twist-1和N-cadherin表达上调,E-cadherin表达下调,而当上调miRNA-186的表达时,Twist-1和N-cadherin表达下调,E-cadherin表达上调。结论在胃癌组织中miRNA-186的表达水平降低,可能通过靶向调控Twist-1的表达,改变E-cadherin和N-cadherin的表达水平,导致胃癌细胞上皮间质转化,进而促进胃癌细胞的侵袭、迁移。     

沉默piRNA-651抑制骨肉瘤细胞系U20S增殖和转移的影响

目的 研究非编码小RNA piRNA-651对骨肉瘤U20S细胞增殖、侵袭与转移的影响.方法 通过RNA干扰技术沉默U20S细胞中的piRNA-651,并应用实时定量PCR（qRT-PCR）检测骨肉瘤U20S细胞中piRNA-651的表达情况;流式细胞术检测细胞周期的变化,MTT实验检测沉默piRNA-651表达对细胞增殖的影响,Transwell侵袭实验检测细胞侵袭能力的变化,细胞划痕实验检测细胞转移能力的变化.结果 piRNA-651-siRNA可有效沉默piRNA-651在骨肉瘤细胞U20S的表达,敲低U20S细胞中piRNA-651的表达可以明显阻滞骨肉瘤细胞的细胞周期,抑制骨肉瘤细胞的增殖、侵袭与转移能力.结论 piRNA-651在骨肉瘤的发生发展及侵袭转移中发挥着重要作用.     

长链非编码RNA MEG3靶向Rac1抑制宫颈癌细胞增殖、迁移的实验研究

目的 检测长链非编码RNA MEG3在宫颈癌细胞中的表达，检测MEG3表达对HeLa细胞增殖、迁移的影响及可能分子机制。方法 采用荧光定量PCR（QPCR）检测正常宫颈上皮细胞HcerEpic和宫颈癌细胞系C-33A、C4-1、Caski、SiHa和HeLa中MEG3的表达； MEG3过表达质粒（MEG3）、阴性对照质粒（NC组）、MEG3+Rac1过表达质粒（MEG3+Rac1组）分别转染HeLa细胞，MTT法检测细胞增殖情况， Transwell实验检测细胞迁移能力，Western blotting检测PCNA、E-cadenin、N-cadenin、Vimentin、Rac1 蛋白表达。结果 QPCR结果显示，C-33A、C4-1、Caski、SiHa和HeLa细胞中MEG3表达水平分别为0.37±0.044、0.65±0.075、0.41±0.071、0.71±0.053和0.42±0.081，均低于HcerEpic细胞的1.02±0.064，差异具有统计学意义（P＜0.05）。MTT法结果显示MEG3组HeLa细胞增殖活力显著低于NC组； MEG3组的迁移细胞数为385±14，显著低于NC组的594±16（P＜0.05）；与NC组比较，MEG3组PCNA、N-cadenin和Vimentin表达下调，E-cadenin表达上调。与MEG3组比较，MEG3+Rac1组显著上调Rac1蛋白表达，上调HeLa细胞增殖活力，增加迁移细胞数。结论 LncRNA MEG3可能通过靶向Rac1信号通路抑制宫颈癌细胞增殖、迁移。