20 W微波照射骨骼肌卫星细胞的增殖与分化：兔骨折内固定模型研究

BACKGROUND:Low power microwave irradiation has been shown to promote the healing of fractures with internal fixation; however, its action mechanisms on the skeletal muscle around the fracture site are unclear. OBJECTIVE:To study the effects of low power microwave irradiation (20 W) on the proliferation ability of skeletal muscle satellite cells in a rabbit model of femoral fracture with internal fixation. METHODS:Forty male New Zealand rabbits were used to establish femoral fracture followed by internal fixation models, and then were equally randomized into spontaneous recovery and microwave treatment groups. Low power microwave irradiation (20 W) was given for 30 consecutive days in the microwave treatment group on day 4 after modeling, while no microwave irradiation was given in the spontaneous recovery group. Rabbit thigh muscles adjacent to the implant were obtained to isolate skeletal muscle satellite cells. Immunohistochemical staining, hematoxylin-eosin staining and quantitative RT-PCR were used to evaluate the ability of the proliferation and differentiation of skeletal muscle satellite cells. RESULTS AND CONCLUSON: Hematoxylin-eosin staining showed that there was no significant difference in the morphology and histology of skeletal muscle tissues between the spontaneous recovery and microwave treatment groups. However, the relative mRNA expression of MyoG in the cultured skeletal muscle satellite cells in vitro and the number of α-sarcometric actin-postive cells in the microwave treatment group were significantly increased compared with the spontaneous recovery group (P < 0.05). The proliferative ability of skeletal muscle satellite cells was inhibited at the early stage, but not at the later stage. Our results suggest that low power microwave irradiation (20 W) can promote the proliferation and differentiation of skeletal muscle satellite cells around the implant in a rabbit model of femoral fracture with internal fixation, and thereby confirm the efficacy and safety of low power microwave irradiation for the internal fixation of fractures. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

骨骼肌减少症与运动训练对肌卫星细胞影响的研究现状及展望

BACKGROUND: The phenomenon of atrophy or reduction of muscle, causing degenerative changes of muscle functions, appears along with age. Sports training, in which muscle satellite cells are of great importance, is beneficial to increase in muscle mass and improvement of muscle function.  OBJECTIVE:To summarize regulatory mechanism of satellite cells in skeletal muscle mass; changes of satellite muscle cells in the degenerative process of muscle mass and strength; declining and reverse effects of sports training intervention; situations and problems of current research and prospective of the future.  METHODS: A computer-based online search was conducted in PubMed database by using the key words of “sarcopenia, skeletal muscle, satellite cells” from 1986 to 2015. The language was limited to English. The eligible papers were further analyzed and reviewed. RESULTS AND CONCLUSION: A total of 168 papers were screened. Finally, 39 papers were selected according to the titles and objectives. Skeletal muscle atrophy is shown as II type muscle fiber atrophy, and the II type muscle fiber satellite cell content decreases simultaneously. Exercise is beneficial to increase muscle mass and improve muscle function in older people. Both resistance and endurance trainings can increase the skeletal muscle, especially the II muscle fiber satellite cell content with a further increase in the satellite cell activation and proliferation. The number and activation degree of satellite cells are related to muscle aging, and satellite cells and proliferation factors regulate muscle cell formation. Therefore, future researches should not only focus on the increase of satellite cell bank, but also explore effective ways to promote the activation of satellite cells, such as exercise training, nutrition and drugs. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

细胞因子对骨骼肌细胞和胰岛β细胞的作用

BACKGROUND: A variety of cytokines such as cytokines, growth factors and inflammatory proteins play an important role in the development of skeletal muscle. OBJECTIVE:To investigate the biological characteristics of a variety of cytokines and their effects on skeletal muscle cells and pancreatic β cells. METHODS: Relevant articles published from 2002 to 2015 were retrieved in CNKI and PubMed databases using the English keywords “cytokines, adiponectin, leptin, visfatin, skeletal muscle cells, pancreatic β cells”. Initially 253 literatures were obtained, and finally 53 eligible literatures were included based on the exclusion criteria. RESULTS AND CONCLUSION: As a fat-specific protein newly found, adiponectin can improve the insulin sensitivity by promoting glucose uptake, storage and utilization in skeletal muscle cells. The activation of muscle satellite cells and skeletal myoblast proliferation are both dependent on leptin, so leptin plays a vital role in the skeletal muscle cell growth and development. Visfatin, a pleiotropic cytokine, widely presents in the skeletal muscle, liver and bone marrow, and participates in the regulation of inflammation and immune function. Furthermore, visfatin contributes to glucose uptake and metabolism in the skeletal muscle, and makes considerable effects on the stress and signal transduction of skeletal muscle cells. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

骨骼肌生长因子、肌球蛋白及骨骼肌胶原与骨骼肌的损伤与修复

BACKGROUND: Regeneration and repair of injured skeletal muscle are influenced by a variety of factors. Skeletal muscle myosin heavy chain and skeletal muscle collagen content are known to be involved in the repair of injured skeletal muscle. OBJECTIVE: To summarize the changes and roles of skeletal muscle growth factors, myosin, and collagen in the repair of injured skeletal muscle. METHODS: A computer-based online search was conducted in PubMed and Wanfang databases from 2002 to 2015 to screen the relevant literatures. Roles of skeletal muscle growth factors, myosin, and collagen in the repair of injured skeletal muscle was summarized by collecting the data regarding the factors influencing exercise and the repair of injured skeletal muscle, and growth factors involved in the regeneration and repair of injured skeletal muscle. RESULTS AND CONCLUSION: Insulin-like growth factor, fibroblast growth factor and hepatocyte growth factor regulate inflammation after skeletal muscle injury extensively. Myosin heavy chain is considered as a specific marker for skeletal muscle regeneration. Types I and III collagen and fibronectin functioning as a skeleton for skeletal muscle fiber play critical roles in the regeneration and repair of injured skeletal muscle. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

生长因子对骨骼肌损伤与修复的作用

BACKGROUND: Growth factors involved in the regulation of cellular processes play an important role in the wound healing and tissue regeneration. OBJECTIVE:To elaborate the role of a variety of cellular processes involving growth factors in the repair of skeletal muscle injury, and to provide the references for the treatment and rehabilitation strategies, and the synthesis of biomaterials with growth factor for the skeletal muscle after injury. METHODS: A computer-based online search was conducted in PubMed, Mendeley, Google Scholar, and CNKI databases from 1995 to November 2015 to screen the relevant literatures using the keywords “skeletal muscle, damage repair, insulin-like growth factor, epidermal growth factor, growth factor”. Data screening, processing, and summary were performed. RESULTS AND CONCLUSION: Fifty-one eligible literatures were included. Exercise training promotes the repair and regeneration of the injured skeletal muscle cells and the recovery of the function by activating satellite cells in the sarcolemma and basement membrane to produce the numerous myoblasts. The repair involves the complex biological process regulated by growth factors. Exogenous growth factors up-regulate the mRNA expression of endogenous growth factors, stimulate the proliferation of the myoblasts, accelerate the fusion between myotubes and muscle fibers, promote the repair of skeletal muscle injury, inhibit the formation of scars, thereby enhancing the healing quality. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Platelet-rich plasma combined with naringin induces osteogenic differentiation of human bone marrow mesenchymal stem cells In vitro北大核心

BACKGROUND: Platelet-rich plasma (PRP) and naringin can both promote proliferation and induce osteogenic differentiation of mesenchymal stem cells. However, their combined use is rarely reported. OBJECTIVE: To observe the effect of PRP combined with naringin on the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) in vitro. METHODS: BMSCs at passage 3 were divided into four groups: (1) blank control group, cells were cultured in α-MEM; (2) PRP group, cells were cultured in α-MEM containing PRP; (3) naringin group, cells were cultured in α-MEM containing naringin; and (4) combined group, cells were cultured in a-MEM containing PRP and naringin. The contents oO used PRP and naringin were 12.5% and 50 µg/L respectively. Cell proliferation was detected by MTT assay. Expression oO related genes in hBMSCs was detected by RT-PCR. Alkaline phosphatase staining, collagen type I immunohistochemical staining, and alizarin red staining were used to analyze the osteogenic differentiation of hBMSCs. RESULTS AND CONCLUSION: The proliferation of hBMSCs was increased in each group, especially in the combined group. Cells in all the groups oxcept the blank control group were positive for alkaline phosphatase staining, collagen type I immunohistochemical staining, and alizarin red staining, and the positive effect was more obvious in the combined group. However, negative or weakly positive response was found in the blank control group. At 7 and 14 days, the expression of alkaline phosphatase and collagen type I was significantly higher in the PRP, naringin and combined groups than the blank control group (P < 0.05); at 14 days, the expression of alkaline phosphatase and collagen type I was significantly higher in the combined group than the PRP and naringin groups (P < 0.05). To conclude, PRP combined with naringin can promote the proliferation of hBMSCs and induce the osteogenic differentiation of hBMSCs. Moreover, there is a synergistic effect between PRP and naringin. © 2018, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.     

制备小鼠去细胞拟胚体对Lewis肺癌细胞的促分化作用

BACKGROUND: Co-culture with embryonic stem cells or embryonic tissues can induce differentiation of carcinoma cells into normal epithelial cells or decrease malignancy of carcinoma cells. Acellular embryoid bodies retain the structure and important cytokines of embryonic tissues. OBJECTIVE:To prepare acellular embryoid bodies from mouse embryonic stem cells and to investigate their effects on differentiation of mouse Lewis lung carcinoma cells at three-dimensional culture in vitro. METHODS: Mouse embryonic stem cells (D3) were dynamically cultured for 7 days to produce embryoid bodies followed by decellularization with 0.1% sodium dodecyl sulfate. Mouse Lewis lung carcinoma cells were co-cultured with acellular embryoid bodies as test group or cultured in three-dimensional matrigel medium for 7 days as control group, respectively. Cell proliferation and expression of E-cadherin were detected by immunohistochemical staining and western blot assay, respectively. In addition, mRNA expressions of Slug and E-cadherin were observed using RT-PCR technology. RESULTS AND CONCLUSION: Uniform mouse embryoid bodies were successfully prepared, and were completely decellularized with sodium dodecyl sulfate. After 7-day three-dimensional matrigel culture, in the control group, multicellular tumor spheroids were formed, accompanied by a higher Ki67 positive rate; Lewis lung carcinoma cells in the test group were repopulated in the acellular embryoid bodies showing significantly lower Ki67 positive rate. Compared with the control group, the absorbance of Paxillin in the test group was significantly smaller, and the absorbance of E-cadherin was significantly higher (P < 0.05). Besides, mRNA expressions of Slug and E-cadherin were significantly decreased and increased in the test group compared with the control group, respectively (P < 0.05). These findings indicate that the acellular embryoid bodies can promote differentiation of mouse Lewis lung carcinoma cells in three-dimensional culture in vitro. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松; ORCID: 组织工程0000-0003-2685-4218(吕卫东)     

人类脂肪来源成体干细胞体外定向成软骨的诱导实验

Objective To investigate the feasibility of inducing human adipose derived adult stem cells (hADASc) into chondrocytes by gene transfection.Methods hADASc were cultured in vitro, and transfected with gene PGL3-TGF-pl, whose Luceferase activity (RUL value) was measured.PT-PCR was used to monitor the expression of tranfected hADASc.Anti collagen II immunohistochemical staining was performed in the experimental group (transfected hADASc) , active control group (untransfected hADASc cultured in chondrocytes induction medium) and negative control group (untransfected hADASc cultured in conventional medium).The immuno-staining image was analyzed with an automated imaging analysis system and computed for PU values.Results Nucleated tissue cells with features of stem cells were isolated from in vitro culture of human subcutaneous adipose tissue.The RUL value of the experimental group (9 212.583±315.240) was higher than that of the control group(317.000?0.710, P＜0.01).RT-PCR revealed that the transfected hADASc could express the TGF-β1.Anti collagen Ⅱ immunohistochemical staining was positive in experimental group and active control group, with PU values (13.864±2.416 and 13.637±2.548,respectively) statistically different from that of the negative control group (6.013 ±0.827, P＜0.05).Conclusions Nucleated tissue cells with features of stem cells can be isolated from culture of human subcutaneous adipose tissue in vitro.The PGL3-TGF-β1-transfected hADASc may express TGF-β1.The endogenous TGF-β1-induced hADASc produces collagen Ⅱ , with similar actions of exogenous TGF-β1.     

碱性成纤维细胞生长因子基因转染骨髓间充质干细胞在失神经肌肉萎缩中的应用

BACKGROUND: How to avoid denervated muscular atrophy is a key factor to improve the therapeutic efficacy on peripheral nerve injuries.OBJECTIVE: To study the effect of basic fibroblast growth factor (bFGF) gene-transfected bone marrow mesenchymal stem cells against denervated muscle atrophy.METHODS:bFGF genes were transfected into rat bone marrow mesenchymal stem cells using viral transfection method, and then MTT, immunohistochemical staining, hematoxylin-eosin staining, RT-PCR, western blot, and ELISA methods were used to detect the transfection efficiency and product expression. Thirty-two Sprague-Dawley rats were selected to make animal models of sciatic nerve injury, and subjected to multi-point intramuscular injection of bFGF-transfected bone marrow mesenchymal stem cells (experimental group) or cell culture fluid (control group). At 2, 4, 6, 8 weeks after transfection, the gastrocnemius muscle tissues were harvested to detect action potential, residual wet weight, and cross-sectional area of muscle fibers.RESULTS AND CONCLUSION: The bFGF gene was successfully transfected into bone marrow mesenchymal stem cells using the viral transfection method. The residual wet weight, cross-sectional area and residual action potential of the gastrocnemius muscle were significantly better in the experimental group than the control group (P < 0.05). These findings indicate that bFGF gene-transfected bone marrow mesenchymal stem cells transplanted into the denervated muscle can retard the development of muscle atrophy.  中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

一种新生SD大鼠皮质源性神经元的培养方法

BACKGROUND: Primary culture in vitro of neurons plays an important role in the development, regeneration, signal transduction mechanisms, neuropharmacology and gene expressions of the nervous system. OBJECTIVE: To establish a simple method for primary culture of high-purity cortical neurons in neonatal Sprague-Dawley rats. METHODS: Cortical tissues were acquired from neonatal Sprague-Dawley rats born 1 day. In traditional experimental group, the whole cortex was removed; in improved experimental group, the cortical tissues, 2-3 mm thick on the brain surface were removed. Single cell suspensions were prepared after papain digestion and centrifugation and were then seeded onto 24-well culture plates containing neuron solutions for primary culture (1×105 per well). Cells were identified by neuronal specific markers MAP-2 and Tuj1 after 3-day culture. The number of neurons and neurite length were observed under inverted phase contrast microscope and recorded at 6, 24, 48 and 72 hours, 5 and 7 days of culture, resprctively. RESULTS AND CONCLUSION:The cultured cells expressing MAP-2 and Tuj1 were neurons that could be used in the following experiments. The purity of neurons in the improved experimental group was 92% at 3 days, while only 51% in the traditional experimental group. Cells in both two groups had attached to the wall presenting with small processes at 6 hours, and a simple neural network formed at the 3rd day until dense neural networks could be found at the 5th day. To conclude, our culture method herein is simple and convenient, and can be used to produce neurons with high purity, which will be helpful for the experimental studies on cortical neurons from Sprague-Dawley rats. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程