miR-3074-5p及其靶基因 p27在子痫前期胎盘组织中的异常表达

目的:观察miR-3074-5p及其靶基因 p27在子痫前期胎盘组织中的表达情况,初步探讨miR-3074-5p/ p27分子途径在子痫前期病理过程中的作用。 方法:收集2017年9月至2018年3月期间在天津医科大学第二医院产科行剖宫产手术的子痫前期患者（子痫前期组,16例）和相同孕...     

miR-485-5p靶向Toll样受体4调控巨噬细胞的炎症反应

目的 探究miR-485-5p对Toll样受体4(TLR-4)的靶向作用，研究其在巨噬细胞炎症反应中的调控作用。方法 合成miR-485-5p模拟物(mimics)及其抑制剂(inhibitors),分别转染HEK-293T细胞，RT-PCR检测miR-485-5p的表达水平。根据miRNA靶向作用预测数据分析选定TLR-4作为研究的靶基因，构建TLR-43’UTR及其突变体萤火虫-海肾双荧光素酶报告基因系统，通过检测在细胞模型中过表达/抑制表达miR-485-5p后对荧光素酶活力的影响，验证miR-485-5p与TLR-4之间的靶向关系，Western blot检测TLR-4蛋白质表达水平。构建巨噬细胞模型，将miR-485-5p mimics及inhibitors分别转染巨噬细胞，RT-PCR检测靶基因TLR-4 mRNA的表达水平，ELISA检测上清中下游致病因子IL-6、 IL-17、IL-18的表达。结果 与对照组比较，转染miR-485-5p mimics后miR-485-5p表达水平升高，转染miR-485-5p inhibitors后miR-485-5p表达水平降低(P...     

多发性骨髓瘤患者血清中miR-223-3p和miR-140-5p表达水平与肾功能损害的相关性分析

目的 探讨血清miR-223-3p和miR-140-5p表达水平与多发性骨髓瘤患者肾功能损害的相关性。方法 选取本院于2019年7月—2022年6月收治的165例多发性骨髓瘤患者,按照入院时诊断结果中存在肾损伤与未见肾损伤分为观察组与对照组。比较2组血清中血肌酐(Scr)、血尿素氮(BUN)、24 h尿蛋白水平以及胱抑素C(Cys-C),检测血清miR-223-3p和miR-140-5p表达水平,分析血清miR-223-3p和miR-140-5p表达水平与多发性骨髓瘤患者肾功能损害的相关性。结果 与对照组比较,观察组患者血清miR-223-3p和miR-140-5p表达水平均降低(P<0.05),血清中Scr、BUN、24 h尿蛋白以及Cys-C水平升高(P<0.05);不同性别、年龄、肿瘤细胞形态、患者血清miR-223-3p、miR-140-5p表达水平差异无统计学意义(P>0.05),低肿瘤分化及与Ⅲ期患者血清miR-223-3p、miR-140-5p表达水平低于高、中分化及Ⅰ期、Ⅱ期患者(P<0.05);Pearson相关性分析结果显示,血清miR-22...     

25-羟基维生素D_3诊断子痫前期的价值

目的检测孕早期女性外周血25-羟基维生素D_3[25(OH)D_3]水平,探讨25(OH)D_3水平与子痫前期的相关性,旨在为子痫前期的早期临床预测、预防及治疗提供理论依据。方法选取2014年5月-2016年6月于湖北医药学院附属人民医院进行产前检查和分娩的781例孕妇作为研究对象,根据妊娠结局及实验室指标分为正常妊娠组(680例)、妊娠期高血压组(45例)、轻度子痫前期组(30例)和重度子痫前期组(26例)。采用离子比色法检测血清钙水平,采用电化学发光免疫分析法检测血清25(OH)D_3水平。制作ROC曲线,评价两者预测子痫前期的价值。结果与正常妊娠组比较,妊娠期高血压组孕妇血清钙和25(OH)D_3水平差异均无统计学意义(均P0.05),轻度子痫前期组和重度子痫前期组孕妇血清钙和25(OH)D_3水平均显著降低(均P0.05)。与妊娠期高血压组比较,轻度子痫前期组和重度子痫前期组孕妇血清钙和25(OH)D_3水平均显著降低(均P0.05)。与轻度子痫前期组比较,重度子痫前期组孕妇血清钙和25(OH)D_3水平均显著降低(均P0.05)。血清钙水平预测子痫前期的ROC曲线下面积为0.76,当其截断值为0.91时,灵敏度为72.52%,特异度为84.94%。血清25(OH)D_3水平预测子痫前期的ROC曲线下面积为0.87,当其截断值为1.41时,灵敏度为78.81%,特异度为87.54%。结论孕妇血清25(OH)D_3水平降低与子痫前期发病相关,可能预示子痫前期的患病风险,对子痫前期有一定的预测价值。     

miR-30c-5p通过调节Notch1的表达抑制人绒毛膜滋养层细胞HTR-8/SVneo的迁移、侵袭和管状形成

目的：探讨miR-30c-5p通过调节Notch1表达对人绒毛膜滋养层细胞HTR-8/SVneo迁移、侵袭和管状形成调控的机制,确定其在子痫前期发病中的潜在作用。方法：在体外对人绒毛膜滋养层细胞HTR-8/SVneo进行培养,慢病毒系统构建miR-30c-5p过表达及敲低稳转HTR-8/SVneo细胞系。分为对照组、导入miR-30c-5p过表达质粒组和导入miR-30c-5p敲低质粒组。通过荧光定量PCR(qRT-PCR)检测各组细胞中miR-30c-5p和Notch1基因的表达量,蛋白质免疫印迹法(Western blot)检测各组细胞中Notch1蛋白的表达量,划痕愈合实验、Transwell侵袭实验和管状形成实验分别检测各组细胞的迁移、侵袭和管状形成能力。结果：与对照组比较,miR-30c-5p过表达质粒组中细胞的miR-30c-5p基因表达水平升高(P<0.0001),细胞Notch1蛋白表达水平下降(P<0.01),细胞迁移(P<0.01)、侵袭(P<0.001)和管状形成能力(P<0.05)均受到抑制。反之,miR-30c-5p敲低质粒组中细...     

CircPAPPA、miR-3127-5p表达与子痫前期妊娠结局的相关性分析

目的 探讨子痫前期(PE)患者血清环状RNA妊娠相关蛋白A(CircPAPPA)、微小RNA-3127-5p(miR-3127-5p)表达水平及与子痫前期妊娠结局相关性。方法 选择2020年6月至2022年3月期间在西北妇女儿童医院产前检查的93例PE孕妇为研究对象,并作为PE组,其中轻度组53例,重度组40例。根据妊娠结局再将93例PE患者分为良好组与不良组。另选择同期健康孕妇90例为对照组。分析比较各组血清CircPAPPA、miR-3127-5p表达水平与临床指标的相关性。结果 PE组剖宫产比例、舒张压、收缩压、24h尿蛋白显著高于对照组,分娩孕周显著低于对照组(χ^(2)/t值分别为6.662、18.409、17.345、29.530、6.573,P<0.05);与对照组比较,PE组患者血清CircPAPPA表达水平显著降低,miR-3127-5p表达水平显著升高(t值分别为10.862、15.749,P<0.05)。与轻度组比较,重度组患者血清CircPAPPA表达水平显著降低,miR-3127-5p表达水平、24h尿蛋白显著升高(t值分别为7.774、9.652、18.493,P<0.05)。相关性分析显示,PE患者血清CircPAPPA与舒张压、收缩压、24h尿蛋白呈负相关(r值分别为-0.475、-0.603、-0.561,P<0.05),而血清miR-3127-5p与舒张压、收缩压、24h尿蛋白呈正相关(r值分别为0.505、0.591、0.607,P<0.05);CircPAPPA与miR-3127-5p表达水平呈负相关(r=-0.610,P<0.05)。与良好组比较,孕中、晚期不良妊娠结局孕妇的血清CircPAPPA表达水平显著降低,miR-3127-5p表达水平显著升高(孕中期:t值分别为2.582、4.407;孕晚期:t值分别为6.095、6.501,P<0.05)。结论 PE患者血清CircPAPPA表达水平降低,miR-3127-5p表达水平升高,且与PE病情和妊娠结局相关,可能成为评估PE病情及妊娠结局的标志物。     

卵巢癌患者血清miR-124-3p和miR-150检测的临床意义分析

目的探讨卵巢癌患者血清miR-124-3p和miR-150检测水平变化情况及临床意义。方法选取2016年1月-2017年1月该院肿瘤科收治的68例卵巢癌患者作为研究组,另选取同期在该院体检健康者68例作为对照组。采用实时荧光定量PCR (qRT-PCR)方法检测所有受试者血清miR-124-3p和miR-150表达水平,分析二者表达与卵巢癌病理特征之间的关系,进一步分析二者对卵巢癌患者生存时间及预后的影响。结果研究组血清miR-124-3p和miR-150表达水平明显低于对照组(P0. 05); miR-124-3p和miR-150表达与年龄、病理类型、肿瘤直径均无相关性(P0. 05),与FIGO分期、肿瘤分级、淋巴结转移密切相关(P0. 05); miR-124-3p高表达组预后5年存活率为40. 54%,低表达组预后5年存活率为22. 58%,高表达组存活率高于低表达组(χ2=4. 541,P=0. 033); miR-150高表达组预后5年存活率为42. 86%,低表达组预后5年存活率为27. 27%,高表达组存活率高于低表达组(χ2=4. 669,P=0. 031);年龄与卵巢癌预后无关(P 0. 05); FIGO分期、肿瘤分级、淋巴结转移、miR-124-3p、miR-150是影响卵巢癌患者预后的危险因素(P0. 05)。结论 miR-124-3p和miR-150在卵巢癌患者血清中均表达下调,并与FIGO分期、肿瘤分级、淋巴结转移有关,可作为卵巢癌患者预后的检测指标。     

子痫前期胎盘组织中LAMA4蛋白表达量及其与滋养细胞侵袭、胎盘缺氧的相关性

目的探讨子痫前期胎盘组织中层粘连蛋白α4链(LAMA4)蛋白表达量及其与滋养细胞侵袭、胎盘缺氧的相关性。方法收集2012年5月-2016年5月在该院接受治疗及分娩的子痫前期患者56例作为观察组;另选取同期进行产检及分娩的正常产妇50例作为正常对照组。根据胎盘组织中LAMA4蛋白表达中位数,观察组患者被分为高LAMA4蛋白组和低LAMA4蛋白组,每组各28例。采用Western-Blot法检测胎盘组织中LAMA4蛋白表达量;采用荧光定量PCR法检测胎盘组织中滋养细胞侵袭相关基因mRNA表达量;采用化学发光法检测脐血中胎盘缺氧相关指标含量。结果观察组胎盘组织LAMA4蛋白表达量低于正常对照组(P0.05);高LAMA4蛋白组、低LAMA4蛋白组胎盘组织中基质金属蛋白酶-9(MMP-9)、血管内皮生长因子(VEGF)、胎盘生长因子(PLGF)mRNA表达量低于正常对照组,可溶性fms-样酪氨酸激酶受体-1(sFlt1)mRNA表达量高于正常对照组,脐血胎盘缺氧指标,缺氧诱导因子-1α(HIF-1α)、缺氧诱导因子脯氨酸羟化酶-1(HPH-1)、瘦素含量均高于正常对照组(P0.05);高LAMA4蛋白组胎盘组织中MMP-9、VEGF、PLGFmRNA表达量高于低LAMA4蛋白组,sFlt1mRNA表达量低于低LAMA4蛋白组,脐血胎盘缺氧指标HIF-1α、HPH-1、瘦素含量低于低LAMA4蛋白组(P0.05)。结论子痫前期胎盘组织中LAMA4蛋白表达量减少,具体LAMA4蛋白表达水平与滋养细胞侵袭呈正相关,与胎盘缺氧程度呈负相关。     

miR-142-3p抑制甲状腺癌细胞增殖的研究

目的 研究甲状腺癌细胞株中miR-142-3p的表达水平,探讨miR-142-3p不同表达水平对甲状腺癌细胞增殖能力的影响.方法 采用RT-PCR检测人甲状腺癌细胞K1、TPC-1、BCPAP和人甲状腺细胞Nthy-ori 3-1中miR-142-3p的表达水平.观察K1细胞分别转染miR-142-3p mimics,...     

Drug-induced modifications of the immune response 8. N-(Substituted phenyl)-2,5-dihydro-2-oxo-4-[(substituted phenyl) amino]-3-furancarboxamide derivatives

The synthesis and anti-allergic activity of a series of novel N-(substituted phenyl)-2,5-dihydro-2-oxo-4-[(substituted phenyl)amino]-3-furancarboxamide derivatives (12) are described. The latter were obtained through a novel, and heretofore unknown rearrangement of 2-(N-phenylamino)-4,5-dihydro-4-oxo-3-furancarboxylic acids 8a to the 2(5H)-furanone amides 12.     

Silk peptides inhibit adipocyte differentiation through modulation of the Notch pathway in C3H10T1/2 cells

Silk protein is a biocompatible material that has been used in many biotechnological applications and exhibits body fat-lowering effects. Recent studies have shown that silk peptides increase expression of osteogenic markers in osteoblast-like cells. Because osteogenic and adipogenic differentiation from common mesenchymal progenitor cells are inverse processes and often regulated reciprocally, we hypothesized that silk peptides might suppress adipocyte differentiation. We therefore endeavored to evaluate the effects of silk peptides on adipocyte differentiation in C3H10T1/2 cells. We find that silk peptides inhibit lipid accumulation and morphological differentiation in these cells. Molecular studies show that silk peptides block expression of adipocyte-specific genes such as peroxisome proliferator-activated receptor γ and its targets, including aP2, Cd36, CCAAT enhancer binding proteinα. Silk peptides appear to inhibit adipogenesis by suppression of the Notch pathway, repressing the Notch target genes Hes-1 and Hey-1. In addition, these peptides inhibit endogenous Notch activation, as shown by a reduction in generation of Notch intracellular domain. N-[N-(3.5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butylester, compound E, and WPE-III-31C, which are all known Notch signaling inhibitors, block adipocyte differentiation to an extent similar to silk peptides. Together, our data demonstrate that silk peptides can modulate adipocyte differentiation through inhibition of the Notch signaling and further suggest potential future strategies for treating obesity and its related metabolic diseases.     

Immune responses elicited by a fourth dose of the HPV-16/18 AS04-adjuvanted vaccine in previously vaccinated adult women

Background

Vaccines are now available for the prevention of HPV-16/18-related cervical infections and pre-cancers, primarily targeting adolescent girls. Since the risk of HPV exposure potentially persists throughout a woman's sexual life, vaccine-derived immunity should be long-term. The current study, HPV-024 (NCT00546078, http://clinicaltrials.gov), assessed the immune memory in North American women who received three doses of HPV-16/18 AS04-adjuvanted vaccine 7 years earlier in HPV-001 (NCT00689741).

Methods

Women vaccinated in HPV-001 received a 4th-dose of the HPV-16/18 vaccine (024-4DV group, N = 65). Post 4th-dose immune responses were compared with post 1st-dose immune responses in cross-vaccination controls (024-3DV group, N = 50). Reactogenicity was compared between the 4th-dose and the 1st-dose administration.

Results

Pre 4th-dose, 100% of subjects in the 024-4DV group remained seropositive for anti-HPV-16/18 antibodies (ELISA). Compared to pre 4th-dose, GMTs for anti-HPV-16 and anti-HPV-18 antibodies were respectively 9.3-fold and 8.7-fold higher at day 7, and 22.7-fold and 17.2-fold higher at month 1. Compared to post 1st-dose, GMTs for anti-HPV-16 and anti-HPV-18 were respectively 80.5-fold and 205.4-fold higher at day 7, and 11.8-fold and 20.5-fold higher at month 1. Furthermore, 68.2% and 77.3% of women had HPV-16/18 specific memory B-cells, respectively, pre 4th-dose, rising to 100% one month post 4th-dose vaccination. The 4th-dose was generally well tolerated.

Conclusion

A 4th-dose of HPV-16/18 AS04-adjuvanted vaccine triggered a rapid and strong anamnestic response in previously vaccinated women, demonstrating vaccine-induced immune memory.