Resolvin D1 attenuates imiquimod-induced mice psoriasiform dermatitis through MAPKs and NF-κB pathways

Background

Resolvin D1 (RvD1), a pro-resolution lipid mediator derived from docosahexaenoic acid (DHA), has been described to promote several kinds of inflammatory resolution. However, the effects and anti-inflammatory mechanisms of RvD1 on psoriasis have not been previously reported.

Development and applications of a real-time quantitative RT-PCR method (QRT-PCR) for BRCA1 mRNA

OBJECTIVES: To develop a real-time quantitative RT-PCR method for BRCA1 mRNA and then use it for the study of BRCA1 gene expression in human MCF-7 breast cancer cells after their exposure to antineoplastic agents and gamma irradiation. DESIGN AND METHODS: The developed QRT-PCR method is based on the real-time monitoring of a fluorescein-labeled TaqMan probe, specific for BRCA1 mRNA, during PCR in the LightCycler. A BRCA1 PCR amplicon was purified, quantitated and used as a standard of known concentration for the development and analytical evaluation of the assay. The method was applied to study the alteration of BRCA1 gene expression after exposure to taxol, doxorubicin, 5-fluorouracil, etoposide or gamma irradiation in human MCF-7 breast cancer cells. RESULTS: The developed method is quantitative, highly specific for mRNA and highly sensitive (detection limit of 4 BRCA1 copies per mug of total RNA). We observed a reduction of BRCA1 expression for all antineoplastic agents used, while the gamma irradiated MCF-7 cells had an increase of expression with a peak at the 10 Gy dose. CONCLUSIONS: The developed BRCA1 QRT-PCR method is quantitative, highly sensitive and specific. The proposed method is rapid, automated, and cost effective and can be used to study BRCA1 expression in a variety of clinical samples.     

弓形虫感染家兔血液中虫体的动态观察

目的探明虫体在家兔血液中动态变化情况。方法用RH株弓形虫速殖子105个/只的剂量腹腔感染雄性家兔16只。在家兔感染前后分别采集家兔血液,抗凝处理后,-40℃冻存备用,用实时定量PCR检测弓形虫感染家兔血液中弓形虫DNA,依据对照计算虫体拷贝量,绘制感染后时间与血液中虫体含量的动态曲线图。结果雄兔弓形虫感染后4d血液中即可检测到虫体DNA,血液中虫体含量为217.887×103个/ml,血液中虫体含量在感染后8 d达到高峰期,血液中虫体含量为248.019×103个/ml,然后虫体含量逐渐下降,在感染后121 d雄兔血液中虫体密度为1.45×103/ml。结论弓形虫感染雄兔血液中虫体含量随感染后时间不同而变化。     

Trmt112 gene expression in mouse embryonic development

Mouse Trmt112, the homologous gene of yeast Trm112 (tRNA methyltransferase 11-2), was initially cloned from RIKEN with uncertain function. The yeast TRM112 is now known to play important roles in RNA methylation. Here, we studied the expression of Trmt112 by in situ hybridization and quantitative real-time RT-PCR (QRT-PCR). A higher expression level of Trmt112 was observed in the brain and nervous system by whole mount in situ hybridization from embryonic day 10.5 (E10.5) to E11.5. At later developmental stages E13.5 and E16.5, abundant expression was prominently found in various organs and tissues including developing brain, nervous system, thymus, lung, liver, intestine, kidney, and cartilage. Furthermore, Trmt112 was persistently expressed from E9.5 to E18.5 on whole embryos and highly expressed in multiple organs at E12.5, E15.5 and E18.5 by QRT-PCR. These results showed that Trmt112 gene was highly and ubiquitously expressed during mouse embryonic development, implying that it might be involved in the morphogenesis of diverse organs and tissues and numerous physiological functions.     

长期口服天麻素对SAMP8鼠记忆力及额叶Sst表达的影响

目的:研究长期口服天麻素对SAMP8鼠记忆力的影响及与额叶Sst表达水平的关系。方法:将4月龄雄性SAMP8鼠20只随机分成2组:天麻素治疗组和对照组。治疗组每只鼠平均每日饮10ml含60μg/ml天麻素的蒸馏水,对照组每日仅饮用蒸馏水,直至12月龄。然后,观察两组鼠皮毛、眼睑、运动等基本情况;并通过passive-avoidance-test实验检测两组鼠对疼痛刺激的短期记忆力;同时用QRT-PCR(quantitative real time polymerase c hain reaction)和Western Blot分别检测两组鼠额叶生长抑素(somatostatin,Sst)的转录水平和表达水平。结果:天麻素治疗组小鼠的毛发光泽好、眼睑无炎症、较活跃,而对照组小鼠脱毛明显、部分小鼠眼睑有炎症、运动少;Passive-avoidance-test实验中天麻素治疗组小鼠明室停留时间及进入暗室小鼠的比例与对照组相比具有显著性差别;QRT-PCR结果显示,天麻素治疗组小鼠额叶中Sst转录水平较对照组升高,二者比较差别具有显著性;Western Blot结果显示,天麻素治疗组小鼠额叶中Sst表达水平较对照组明显升高。结论:天麻素治疗可能通过增进额叶Sst的转录和表达水平以改善SAMP8鼠的学习记忆能力。     

Neonatal Osteomacs and Bone Marrow Macrophages Differ in Phenotypic Marker Expression and Function

Osteomacs (OM) are specialized bone-resident macrophages that are a component of the hematopoietic niche and support bone formation. Also located in the niche are a second subset of macrophages, namely bone marrow–derived macrophages (BM Mφ). We previously reported that a subpopulation of OM co-express both CD166 and CSF1R, the receptor for macrophage colony-stimulating factor (MCSF), and that OM form more bone-resorbing osteoclasts than BM Mφ. Reported here are single-cell quantitative RT-PCR (qRT-PCR), mass cytometry (CyTOF), and marker-specific functional studies that further identify differences between OM and BM Mφ from neonatal C57Bl/6 mice. Although OM express higher levels of CSF1R and MCSF, they do not respond to MCSF-induced proliferation, in contrast to BM Mφ. Moreover, receptor activator of NF-κB ligand (RANKL), without the addition of MCSF, was sufficient to induce osteoclast formation in OM but not BM Mφ cultures. OM express higher levels of CD166 than BM Mφ, and we found that osteoclast formation by CD166^−/− OM was reduced compared with wild-type (WT) OM, whereas CD166^−/− BM Mφ showed enhanced osteoclast formation. CD110/c-Mpl, the receptor for thrombopoietin (TPO), was also higher in OM, but TPO did not alter OM-derived osteoclast formation, whereas TPO stimulated BM Mφ osteoclast formation. CyTOF analyses demonstrated OM uniquely co-express CD86 and CD206, markers of M1 and M2 polarized macrophages, respectively. OM performed equivalent phagocytosis in response to LPS or IL-4/IL-10, which induce polarization to M1 and M2 subtypes, respectively, whereas BM Mφ were less competent at phagocytosis when polarized to the M2 subtype. Moreover, in contrast to BM Mφ, LPS treatment of OM led to the upregulation of CD80, an M1 marker, as well as IL-10 and IL-6, known anti-inflammatory cytokines. Overall, these data reveal that OM and BM Mφ are distinct subgroups of macrophages, whose phenotypic and functional differences in proliferation, phagocytosis, and osteoclast formation may contribute physiological specificity during health and disease. © 2021 American Society for Bone and Mineral Research (ASBMR).     

RRBP1在早期发现甲状腺微小癌中的诊断意义

目的观察核糖体结合蛋白1(RRBP1)在早期发现甲状腺微小癌中的诊断意义。方法收集60例甲状腺微小癌、50例甲状腺瘤及50例健康者的静脉血标本,采用实时荧光定量聚合酶链式反应(QRT-PCR)检测RRBP1的表达,并分析RRBP1在甲状腺微小癌中的表达与临床病理特征的关系。结果 RRBP1在甲状腺微小癌中的表达明显高于甲状腺瘤及健康者,差异有统计学意义(P0.05);同时,RRBP1在甲状腺瘤中的表达也明显高于健康者(P0.05)。此外,RRBP1在甲状腺癌患者中的表达与肿瘤大小及淋巴结转移明显相关(P0.05),而与患者的年龄、性别、吸烟史、肿瘤的好发部位无明显相关。结论 RRBP1表达与甲状腺微小癌的发生相关,其表达的检测有助于甲状腺微小癌的诊断。     

高迁移率族蛋白A1(HMGA1)在肝细胞癌中的表达及临床意义

目的:研究肝细胞癌(hepatocelluar carcinoma,HCC)中高迁移率族蛋白A1(high mobility group protein A1,HMGA1)的表达情况,探讨HMGA1基因在HCC中的表达与肝内转移以及临床病理参数之间的关系。方法:采用实时定量荧光聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,QRT-PCR)检测52例肝细胞癌组织和配对的癌旁组织、10例正常肝脏组织中HMGA1mRNA的表达,用免疫组化法(immunohistochemistry,IHC)检测HMGA1蛋白在肝细胞癌中的表达,评价HMGA1的表达与HCC临床病理学因素的相关性。结果:肝癌组织中HMGA1mRNA的表达较正常肝脏中升高6.82倍,其中有、无肝内转移组较正常肝组织分别升高13.36倍和4.63倍(P<0.05)。HMGA1mRNA的表达与TNM分期、Edmonson分级、肝内转移有关(P<0.05)。肝细胞癌中有肝内转移组较无肝内转移组HMGA1蛋白表达明显增强。结论:HMGA1基因在肝细胞癌中的表达较正常肝脏组...