新生鼠胰岛干细胞分离、培养和分化的实验研究

目的　探索新生鼠胰岛干细胞体外分离鉴定、培养以及体外分化的方法。方法　用胶原酶消化新生大鼠胰腺 ,将消化的组织碎片在pH7.4～ 7.6条件下经含血清RPMI 164 0及无血清RPMI164 0 (添加bFGF ,EGF ,N 2 )培养液中培养 ,观察形成类胰岛样细胞团的全过程。用胰岛素释放实验检测胰岛功能 ,免疫荧光法检测nestin的表达。结果　胰腺消化碎片培养 3 6h内 ,可见nestin阳性细胞贴壁 ,添加bFGF ,EGF ,N2后nestin阳性细胞快速增长 ,18～ 2 4d形成类胰岛样细胞团 ,并可表达胰岛素。结论　胰腺细胞中nestin阳性细胞具有胰岛干细胞特点 ,经体外培养可获得类胰岛样细胞团     

Bone From Blood: Characteristics and Clinical Implications of Circulating Osteogenic Progenitor (COP) Cells

Circulating osteogenic progenitor (COP) cells are a population of cells in the peripheral blood with the capacity for bone formation, as well as broader differentiation into mesoderm-like cells in vitro. Although some of their biological characteristics are documented in vitro, their role in diseases of the musculoskeletal system remains yet to be fully evaluated. In this review, we provide an overview of the role of COP cells in a number of physiological and pathological conditions, as well as identify areas for future research. In addition, we suggest possible areas for clinical utilization in the management of musculoskeletal diseases. © 2020 American Society for Bone and Mineral Research (ASBMR).     

Suppression of autophagy by FIP200 deletion leads to osteopenia in mice through the inhibition of osteoblast terminal differentiation

Autophagy is a conserved lysosomal degradation process that has important roles in both normal human physiology and disease. However, the function of autophagy in bone homeostasis is not well understood. Here, we report that autophagy is activated during osteoblast differentiation. Ablation of focal adhesion kinase family interacting protein of 200 kD (FIP200), an essential component of mammalian autophagy, led to multiple autophagic defects in osteoblasts including aberrantly increased p62 expression, deficient LC3‐II conversion, defective autophagy flux, absence of GFP‐LC3 puncta in FIP200‐null osteoblasts expressing transgenic GFP‐LC3, and absence of autophagosome‐like structures by electron microscope examination. Osteoblast‐specific deletion of FIP200 led to osteopenia in mice. Histomorphometric analysis revealed that the osteopenia was the result of cell‐autonomous effects of FIP200 deletion on osteoblasts. FIP200 deletion led to defective osteoblast terminal differentiation in both primary bone marrow and calvarial osteoblasts in vitro. Interestingly, both proliferation and differentiation were not adversely affected by FIP200 deletion in early cultures. However, FIP200 deletion led to defective osteoblast nodule formation after initial proliferation and differentiation. Furthermore, treatment with autophagy inhibitors recapitulated the effects of FIP200 deletion on osteoblast differentiation. Taken together, these data identify FIP200 as an important regulator of bone development and reveal a novel role of autophagy in osteoblast function through its positive role in supporting osteoblast nodule formation and differentiation. © 2013 American Society for Bone and Mineral Research.     

Runx2 Regulates Endochondral Ossification Through Control of Chondrocyte Proliferation and Differentiation

Synthesis of cartilage by chondrocytes is an obligatory step for endochondral ossification. Global deletion of the Runx2 gene results in complete failure of the ossification process, but the underlying cellular and molecular mechanisms are not fully known. Here, we elucidated Runx2 regulatory control distinctive to chondrocyte and cartilage tissue by generating Runx2 exon 8 floxed mice. Deletion of Runx2 gene in chondrocytes caused failure of endochondral ossification and lethality at birth. The limbs of Runx2^ΔE8/ΔE8 mice were devoid of mature chondrocytes, vasculature, and marrow. We demonstrate that the C‐terminus of Runx2 drives its biological activity. Importantly, nuclear import and DNA binding functions of Runx2 are insufficient for chondrogenesis. Molecular studies revealed that despite normal levels of Sox9 and PTHrP, chondrocyte differentiation and cartilage growth are disrupted in Runx2^ΔE8/ΔE8 mice. Loss of Runx2 in chondrocytes also impaired osteoprotegerin‐receptor activator of NF‐κB ligand (OPG‐RANKL) signaling and chondroclast development. Dwarfism observed in Runx2 mutants was associated with the near absence of proliferative zone in the growth plates. Finally, we show Runx2 directly regulates a unique set of cell cycle genes, Gpr132, Sfn, c‐Myb, and Cyclin A1, to control proliferative capacity of chondrocyte. Thus, Runx2 is obligatory for both proliferation and differentiation of chondrocytes. © 2014 American Society for Bone and Mineral Research.     

从体温调节角度研究阳虚恶寒阴虚发热的机理

本文从体温调节角度探讨了阳虚恶寒和阴虚发热的病理机制。畏寒肢冷及五心烦热等现象的出现,可能表明在阳虚或阴虚状态下机体体温调节过程出现某种程度的障碍,并表现为机体对外界冷热刺激的反应能力较正常人低下.鉴于这方面的研究尚不深入,建议从实验室和临床开展工作,有助于阐明阳虚和阴虚证的病理实质以及有助于临床诊断的客观化和标准化。     

脓毒症患者高迁移率族蛋白1水平与卫气营血辨证、APACHEⅡ评分的相关性研究

[目的]探讨不同中医证型脓毒症患者血清高迁移率族蛋白1(HMGB1)水平与急性生理和慢性健康评估Ⅱ(APACHEⅡ)评分、卫气营血辨证分型的相关性.[方法]24例脓毒症患者按卫气营血辨证分为气分证组11例、营血分证组13例,并设8例正常对照组;分别进行APACHEⅡ评分,采用Western blot法检测各组患者血清HMGB1含量,并对HMGB1水平、APACHEⅡ评分及卫气营血辨证证型进行相关性分析.[结果]营血分证组APACHEⅡ评分显著升高,与气分证组比较差异有显著性意义(P＜0.01),且不同辨证分型与APACHEⅡ评分呈正相关(P＜0.01);气分证组、营血分证组患者血清HMGB1水平较正常对照组显著升高(P＜0.05或P＜0.01),而营血分证组血清HMGB1水平较气分证组显著升高(P＜0.01),且不同辨证分型与血清HMGB1水平、APACHEⅡ评分与血清HMGB1水平均呈正相关(P＜0.01).[结论]脓毒症按卫气营血理论辨证,可反映其病理过程及病情严重程度.