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28 April 2017, Volume 21 Issue 12 Previous Issue    Next Issue

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Effect of different resting time on patella-patellar tendon healing after acute injury Liang Xiao-tian1, Wang Bo1, Zeng Xiao-hui1, Tang Yi-ni1, Xiao Fang-xin1, Li Hai-wei2, Wang Lin3 2017, 21 (12):  1805-1812.  doi: 10.3969/j.issn.2095-4344.2017.12.001 Abstract ( 327 )   PDF (1807KB) ( 460 )   Save BACKGROUND: Patella-patellar tendon junction (PPTJ) injuries are a common disease in athletes and vulnerable to re-injury. Post-injury training is a double-edged sword. OBJECTIVE: To examine the effect of post-injury resting days on the quality of healing in PPTJ in view of histomorphology, expression levels of growth factor and collagen, and muscle tension. METHODS: Forty New Zealand white rabbits were randomly divided into control, and five post-injury training groups representing different time to restart training after acute injury: the rabbits restarted training at 24 (PIT24), 48 (PIT48), 72 (PIT72), 96 (PIT96) and 120 (PIT120) hours, respectively. All post-injury training groups received acupuncture at the PPTJ and then underwent 4-week low-intensity training. Afterwards, all rabbits were killed to take specimens for hematoxylin-eosin, safranin O and immuohistochemical staining. The muscle tension and hardness were detected using Myoton-3. The thickness of fibrous cartilage zones, cell density, as well as the expression levels of vascular endothelial growth factor and collagen were measured on Metamorph software. RESULTS AND CONCLUSION: Compared with the control group, in the PIT groups, the collagens arranged in disorder, and decreased cell density, increased mucopolysaccharide and “rising tide” phenomenon appeared. The fibrocartilage in the PIT24 and PIT120 groups was significantly thicker than that in the control group (P < 0.05). The ratio of collagen type I/III was significantly decreased in the PIT24 and PIT48 groups compared with the control group (P < 0.05), and the highest in the PIT72 group. The expression level of vascular endothelial growth factor in the PIT48, PIT72 and PIT96 groups was significantly higher than that in the control group (P < 0.05). The muscle tension was negatively correlated with the collagen ratio (r=-0.538; P < 0.01) and chondrocyte density (r=-0.570; P < 0.01), while muscle hardness was positively correlated with the expression level of vascular endothelial growth factor (r=0.613; P < 0.01). Our results suggest that different resting time makes significant differences in the PPTJ healing and the 72-hour resting is the best choice. Moreover, muscular properties are related to the cell density, collagen synthesis and growth factor expression, which can be used as a non-invasive indicator to guide post-injury training and prevent recurrence. 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 Figures and Tables | References | Related Articles | Metrics

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Association of insulin-like growth factor 1 receptor gene rs2229765 polymorphism with osteoporosis in postmenopausal women Wang Yu1, Guo Tian-kang2, Liu Jing3, Yang Rui-fei3, Shao Fei-fei3, Tian Li-min3 2017, 21 (12):  1813-1818.  doi: 10.3969/j.issn.2095-4344.2017.12.002 Abstract ( 302 )   PDF (1156KB) ( 416 )   Save BACKGROUND: Insulin-like growth factor 1(IGF-1) plays an important role in regulating osteocyte function and bone metabolism. The association of insulin-like growth factor 1 receptor (IGF-1R) gene polymorphisms with osteoporosis in postmenopausal women has not yet been reported. OBJECTIVE: To investigate the correlation between IGF-1R gene rs2229765 single nucleotide polymorphism and osteoporosis in postmenopausal women. METHODS: IGF-1R gene rs2229765 SNPs were detected using PCR-RFLP in 218 patients with postmenopausal osteoporosis and 270 postmenopausal women with normal bone mineral density. The bone mineral density of the lumbar spine, femoral neck and forearm was determined by dual-energy X-ray absorptiometry. Serum IGF-1 level was investigated by ELISA. RESULTS AND CONCLUSION: The AA genotype (29% vs. 17%, P=0.001) and A allele (51% vs. 40%, P=0.000) distributions of the rs2229765 polymorphism in the osteoporosis group were significantly higher than those in the control group. Compared with GG genotype of rs2229765, AA genotype was significantly associated with an increased risk of osteoporosis (OR=2.12, 95%CI=1.27-3.54, P=0.004). The analysis of serum IGF-1 showed that osteoporotic women with rs2229765AA (P=0.007) and GA (P=0.016) genotype were found to have a lower serum IGF-1 level than osteoporotic women with GG genotype. Our results indicate that the IGF-1R gene rs2229765 polymorphism capable of regulating serum IGF-1 level is associated with postmenopausal osteoporosis. 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 Figures and Tables | References | Related Articles | Metrics

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Influence of M2 macrophage supernatant combined with Eucommia flavonoids on the biological behavior of osteoblasts under hypoxic conditions Zhang Wen-bo1, Zhang Xian2 2017, 21 (12):  1819-1825.  doi: 10.3969/j.issn.2095-4344.2017.12.003 Abstract ( 351 )   PDF (5178KB) ( 627 )   Save BACKGROUND: Tissue-engineered materials containing osteoblasts have been widely used in the treatment of orthopedic diseases. How to improve hypoxic tolerance and the biological behavior of osteoblasts under hypoxic conditions is an important goal of the research on tissue engineering. OBJECTIVE: To explore the effect of M2 macrophage supernatant and Eucommia flavonoids on the biological behavior of osteoblasts under hypoxia to pave ways for tissue engineering. METHODS: After MC3T3E1 cells were resuscitated, and M2 macrophages were isolated, induced and cultured, all cells were divided into control, M2 macrophage, Eucommia flavonoids and combination groups. The control group received no intervention. Cells in the M2 macrophage, Eucommia flavonoids and combination groups were exposed to M2 macrophages or/and 100 mg/L Eucommia flavonoids, respectively. RESULTS AND CONCLUSION: The cell viability in the M2 macrophage, Eucommia flavonoids and combination groups was significantly higher than that in the control group. The percentage of cells in the S and G2/M phases in the combination group was significantly higher than that in the control group. The relative mRNA expression levels of Runx2, collagen type I and alkaline phosphatase, alkaline phosphatase activity and relative concentration of calcium ions in the combination group were significantly higher than those in the other three groups. To conclude, M2 macrophage supernatant combined with Eucommia flavonoids can promote osteoblast proliferation and differentiation under hypoxic conditions, maintain normal mineralization and osteogenesis of osteoblasts, and improve the tolerance ability of osteoblasts to hypoxia. 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 Figures and Tables | References | Related Articles | Metrics

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Advanced glycation end products modulate osteoclastic acidification by inhibiting the expression of V-ATPase a3 and ClC-7 Wang Hai-xing1, Li Zi-qing1, Xiao Yin-bo1, Zhang Zi-ji1, Zhang Yang-chun1, Yang Xing1, Li Chao-hong2, Sheng Pu-yi1 2017, 21 (12):  1826-1832.  doi: 10.3969/j.issn.2095-4344.2017.12.004 Abstract ( 363 )   PDF (6813KB) ( 396 )   Save BACKGROUND: The effect of advanced glycation end products (AGEs) on bone resorption is controversial. Our previous study has shown that bone resorption is significantly inhibited when AGEs present with pre-osteoclast cells RAW 264.7, while the effect of AGEs on osteoclastic acidification remains unknown. OBJECTIVE: To investigate the effect of AGEs on osteoclastic acidification and the underlying mechanism. METHODS: RAW 264.7 cells were induced by RANKL (15 μg/L; normal group) to generate osteoclasts, and AGEs  (50-400 mg/L; experimental group) or bovine serum albumin (100 mg/L; control group) were added at the beginning of the induction. The effect of AGEs on bone resorption was evaluated by analyzing the area of bone resorption on the Osteo Assay Surface plates, and the effect of AGEs on osteoclastic acidification was evaluated by acridine orange staining. Furthermore, the expression levels of V-ATPase a3 and ClC-7 were detected to investigate the underlying mechanism. RESULTS AND CONCLUSION: The bone resorption area in the AGEs group was significantly decreased compared with the normal group (P < 0.05). Acridine orange staining revealed that the red fluorescence (620 nm) intensity in the AGEs group was significantly decreased compared with the normal group (P < 0.05), and this inhibitory effect became obvious with the increase of AGEs concentration. Immunocytochemistry, western blot assay and PCR findings showed that the expression levels of V-ATPase a3 and ClC-7 in the AGEs group were decreased significantly compared with the normal group (P < 0.05). To conclude, AGEs exert inhibitory effect on osteoclastic acidification, probably by inhibiting the expression of V-ATPase a3 and ClC-7. 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 Figures and Tables | References | Related Articles | Metrics

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Culture of rat primary osteoblasts using enzymatic digestion combined with tissue explant method Ding Xiang-ying1, Cai Jing-wei2, Pan Ji-ming1, Liang Min2 2017, 21 (12):  1833-1837.  doi: 10.3969/j.issn.2095-4344.2017.12.005 Abstract ( 342 )   PDF (5718KB) ( 431 )   Save BACKGROUND: Osteoblasts with high purity and activity are essential for bone metabolism research. OBJECTIVE: To explore a simple and effective culturing method of primary osteoblasts. METHODS: Osteoblasts were isolated from the parietal and frontal bones of newborn Sprague-Dawley rats using trypsin and collagenase digestion and tissue explant method. The morphology of osteoblasts was observed by inverted phase contrast microscope and transmission electron microscope; the cells was counted to draw the growth curve; the osteoblasts were identified by alkaline phosphatase BCIP/NBT staining and alizarin red staining. RESULTS AND CONCLUSION: The cells showed spindle, triangle or polygon shapes, having two or three protrusions. There were abundant mitochondria and endoplasmic reticulum under electron microscope, which presented the typical characteristics of osteoblasts. The cell growth was slow intially, accelerating at the 3rd day,  and peaking at the 7th day. The cells were highly positive for alkaline phosphatase staining and were stained orangered through the alizarin red staining. To conclude, the cells isolated using enzymatic digestion combined with tissue explant method exhibit the typical characteristics and functions of osteoblasts, and this method is an ideal way to culture primary osteoblasts. 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 Figures and Tables | References | Related Articles | Metrics

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Lipoic acid effects on electrophysiological changes of the sciatic nerve following ischemia/reperfusion injury Fu Ling1, Huang Bo2, Li Yi-lin3, Song Ning4, Mo Yi-nan4, Ma Hong5 2017, 21 (12):  1838-1842.  doi: 10.3969/j.issn.2095-4344.2017.12.006 Abstract ( 306 )   PDF (4034KB) ( 319 )   Save BACKGROUND: Lipoic acid, with a closed circle structure composed by sulphur and carbon atoms, exerts strong anti-oxidation, and has been extensively applied in the prevention and treatment of oxidative stress, diabetic cataract, diabetic neuropathy and cardiovascular diseases. OBJECTIVE: To investigate the protective effect of lipoic acid on peripheral nerve function during peripheral nerve ischemia/reperfusion injury. METHODS: Models of peripheral nerve ischemia/reperfusion injury were established in rabbits, and then rabbit models were then allotted to treatment and non-treatment groups. The treatment group was subdivided into experimental (injection of lippoic acid) and control groups according to the use of lipoic acid at 1, 3 and 6 hours after ischemia and before reperfusion. The ultrastructural changes of the sciatic nerve were observed under electron microscope, and the electrophysiological changes of the sciatic nerve were detected using evoked potential instrument. RESULTS AND CONCLUSION: With the ischemic time increasing, the number of vacuoles in the axon increased gradually, accompanied by axonal atrophy, and Waller’s degeneration in the aggregated microfilaments. The myelin sheath thickening and dissolving were visible. All above phenomena became severest at 6 hours after ischemia. Compared with the control groups, lipoic acid reduced the number of the vacuoles in the axon and alleviated axonal atrophy, Waller’s degeneration and demyelination. As the ischemic time increasing, the latency of sciatic nerve was significantly increased, and peaked at 6 hours of ischemia; while the amplitude was significantly decreased, and reached a minimum at 6 hours of ischemia. Compared with the control groups, in the experimental groups, the latency of sciatic nerve was significantly decreased, but the amplitude was significantly increased. These results suggest that lipoic acid provides neuroprotection against peripheral nerve ischemia/reperfusion injury. 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 Figures and Tables | References | Related Articles | Metrics

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Chondrogenesis of chondrocyte cell sheets Zhou Li-bin, Wu Wei, Ding Rui-ying, Xu Bing-xin, Han Hao-lun, Wang Gang, Li Bao-wei, Wang Hong-nan 2017, 21 (12):  1843-1848.  doi: 10.3969/j.issn.2095-4344.2017.12.007 Abstract ( 428 )   PDF (2475KB) ( 306 )   Save BACKGROUND: The cell-sheet technology, based on a temperature-responsive culture, has been drawing more and more attention; however, the temperature-responsive culture dish is quite expensive. Therefore, it is imperative to develop a substitutive technique.     OBJECTIVE: To study the feasibility of cell-sheet culturing using common culture dish, and investigate the chondrogenesis of the cell sheet. METHODS: A piece of nasal septal cartilage was adopted from a patient with deviation of nasal septum to extract primary chondrocytes that were then cultured and amplified. The passage 3 chondrocytes were used to construct cell sheets. Monolayer cell sheet was formed by intensive culturing and allowing the extracellular matrix secretion. Bilayer cell sheet was constructed by seeding passage 2 chondrocytes on the monolayer cell sheet. The cell sheets were harvested using cell scraper, their properties were investigated prior to implantation into nude mice to construct the tissue-engineered cartilage. RESULTS AND CONCLUSION: Both bilayer and monolayer cell sheets with soft tremellose structures showed no significant difference through naked eyes. The newly harvested cell sheets appeared to have good fluidity and gelation. Eight weeks after implantation into the nude mice, mature cartilage blocks were obtained. Histologically, the cell sheets were thin films composed by layered chondrocytes and extracellular matrix. Glycosaminoglycan formation and type II collagen expressions were observed in the cell sheets cultured in vitro. The explanted samples exhibited mature cartilaginous tissue at 8 weeks after implantation. Biochemical analysis showed that the DNA contents of the neocartilages were higher than those of native human costal cartilage, while the contents of glycosaminoglycan and hydroxyproline were similar to native human nasal septal cartilage. To conclude, the chondrocyte cell sheets are likely to be constructed and harvested successfully using common culture dish, and the cell sheets exhibit favourable chondrogenesis. 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 Figures and Tables | References | Related Articles | Metrics

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Association of COL1A1, COL5A1 and COL12A1 genes with anterior cruciate ligament injury Chen Hong1, Shi Li2, Zhang Jun3, Zhao Dao-hong3, Shi Lei2, Lu Qin-nan1, Zhang Qi1, Duan Qi-hui1, Shu Song-hua1 2017, 21 (12):  1849-1854.  doi: 10.3969/j.issn.2095-4344.2017.12.008 Abstract ( 615 )   PDF (1222KB) ( 451 )   Save BACKGROUND: The etiology of anterior cruciate ligament injury remains unclear yet, and some researchers have pointed that interior and exterior factors both contribute to anterior cruciate ligament injury; additionally, the genetic factor interior factors stand out. Collagen genes COL1A1, COL5A1, and COL12A1 are reported to be associated with anterior cruciate ligament injury in Caucasian populations. OBJECTIVE: To investigate the association of polymorphisms of COL1A1, COL5A1 and COL12A1 genes with anterior cruciate ligament injury in Chinese Han population . METHODS: 105 patients with anterior cruciate ligament injury were enrolled and 110 patients without history of anterior cruciate ligament injury were as controlls. The first intron rs1800012 in COL1A1, rs127722 and rs13946 in the 3’-UTR region of COL5A1 gene, rs970547 and rs240736 in the 65 and 29 regions of COL12A1 extron were detected and classified by restriction fragment length polymorphism and genetic sequencing technology. RESULTS AND CONCLUSION: rs1800012, rs12722 and rs13946 genotypes, phenotypes and haplotypes in COL1A1 and COL5A1 genes showed no significant differences between groups. rs970547 and rs240736 genotypes as well as phenotypes and haplotypes in COL12A1 also showed no significant differences between groups. However, there was a significant difference in rs970547 gene frequence in male patients between groups. In conclusion, the Sp1 binding site of COL1A1 rs1800012 is not the susceptibility locus of anterior cruciate ligament injury in Chinese Han population. COL5A1 genes rs12722 and rs13946 in COL5A1 are not closely related to anterior cruciate ligament injury. COL12A1 rs970547 and rs240736 have a certain association with anterior cruciate ligament injury in Chinese men. Male individuals with COL12A1 rs970547 A allelic gene and AA genotype are likely to be susceptible to anterior cruciate ligament injury in Chinese Han population. 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 Figures and Tables | References | Related Articles | Metrics

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Establishing an intervertebral disc degeneration model using annulus puncture Wang Zi-xing, Wei Peng 2017, 21 (12):  1855-1860.  doi: 10.3969/j.issn.2095-4344.2017.12.009 Abstract ( 493 )   PDF (2120KB) ( 351 )   Save BACKGROUND: Annulus puncture method is commonly reported in the establishment of intervertebral disc degeneration (IDD) models, but there is a lack of comparative studies concerning complete MRI, pathologyical and extracellular matrix changes. Thereafter, a further exploration is imperative. OBJECTIVE: To investigate the feasibility of establishing IDD models using annulus puncture. METHODS: Thirty New Zealand white rabbits were randomly divided into two groups (n=15 per group). In experimental group, through extraperitoneal approach, the L1-2, L2-3, and L3-4 intervertebral space were exposed, and then stabbed by 16-gauge hypodermic needles; while rabbits in sham operated group were only subjected to making and incision but not to puncture. Five rabbits in each group were randomly selected for MRI examination at 4, 8 and 16 weeks after modeling, and were sacrificed through air embolism to obtain the nucleus pulposus. Histological changes were observed by hematoxylin-eosin staining, and the level of proteoglycan was detected by Safranin O staining. RESULTS AND CONCLUSION: In the experimental group, T2-weighted signal in the disc was decreased in a time-dependent manner; an obvious reduction was found at 4 weeks after modeling, and reached the peak at 16 weeks. In contrast, the sham operated group showed no significant changes at each time point. Hematoxylin-eosin staining showed that in the experimental group, cartilage-like cells decreased, while fibroblast-like cells in the disc increased in number and proportion, and extracellular fibers were in disorder; all above changes became obvious at 4 weeks after surgery and the most obvious at 16 weeks, but there were no significant changes in the sham operated group. Safranin O staining revealed that the color of the tissues under puncture became lighter, and this variation became overt at 4 weeks after surgery and the most obvious at 16 weeks. In conclusion, annulus puncture is a reliable method to make IDD models. 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 Figures and Tables | References | Related Articles | Metrics

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Liuwei Dihuang Pill inhibits mitogen-activated protein kinase signaling cascade of nucleus pulposus cells in a model of intervertebral disc degeneration Xu Wu-ji1, Liu Si-hua2, Yang Kang3 2017, 21 (12):  1861-1866.  doi: 10.3969/j.issn.2095-4344.2017.12.010 Abstract ( 325 )   PDF (1064KB) ( 401 )   Save BACKGROUND: JNKs and p38 MAPK, as important members of mitogen-activated protein kinase (MAPK), play critical roles in the cell proliferation, differentiation, transformation and apoptosis. OBJECTIVE: To observe the effects of Liuwei Dihuang Pill on the MAPK signaling cascade of nucleus pulposus cells in a rabbit model of intervertebral disc degeneration. METHODS: 100 discs samples with endplate cartilages were equivalently randomized into five groups and cultured in the medium containing 10 mg/L tumor necrosis factor-α (TNF-α, 2 μL), p38-JNK/SAPK blockers (10 μL of 20 μmol/L SB203580 and SP600125, 2 μL of 10 μmol/L TNF-α), 10% Liuwei Dihuang Pill serum, 10 mg/L TNF-α (2 μL) plus 10% Liuwei Dihuang Pill serum, respectively. Those cells received no intervention as controls. Samples were collected and detected at 2, 4, 8 and 14 days of culture. RESULTS AND CONCLUSION: Real time-PCR and western blot assay showed that mRNA and protein expressions of JNK and P38 pathways by the nucleus pulposus cells were upregulated with time, while all above indicators were significantly decreased by the Liuwei Dihuang Pill serum. These findings reveal that Liuwei Dihuang Pill can delay the process of disc degeneration by blocking JNK and P38 signaling pathways, thus exerting protective effect on the intervertebral discs. 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 Figures and Tables | References | Related Articles | Metrics

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Role and significance of hydrogen peroxide-induced transforming growth factor beta1 expression in ligamentum flavum hypertrophy Wang Zhi-qing, Chen Xiong-sheng, Zhou Sheng-yuan, Xu Guo-feng 2017, 21 (12):  1867-1871.  doi: 10.3969/j.issn.2095-4344.2017.12.011 Abstract ( 320 )   PDF (4598KB) ( 356 )   Save BACKGROUND: The pathogenesis of ligamentum flavum hypertrophy remains poorly understood, and the expression of transforming growth factor beta1 (TGF-β1) is increased notably. Reactive oxygen species (ROS) accumulation is associated with tissue degeneration, which may accelerate the progression of ligamentum flavum hypertrophy by upregulating TGF-β1 expression. OBJECTIVE: To clarify the effect and significance of ROS H2O2-mediated up-regulation of TGF-β1 and collagen type I in the progress of ligamentum flavum hypertrophy. METHODS: Ligamentum flavum was removed from a case of acquired lumbar disc herniation with normal ligamentum flavum during lumbar posterior decompression surgery, and then separated and cultured in vitro to the 4-6 generations, followed by exposure to H2O2 at various concentrations (0, 50, 100, 150, 200 μmol/L) for 72 hours. The mRNA and protein expression levels of TGF-β1 and collagen type I were detected by real-time PCR and western blot assay, respectively. RESULTS AND CONCLUSION: Real-time quantitative PCR showed that the mRNA expression level of TGF-β1 was significantly increased in the 150 and 200 μmol/L groups (P < 0.05). The mRNA expression level of collagen type I was significantly higher in the experimental groups than that in the control group, especially in the 200 μmol/L group (P < 0.05). Western blot assay revealed that the protein expression levels of TGF-β1 and collagen type I were significantly increased in a dose-dependent manner (P < 0.05). These findings indicate that H2O2 may accelerate the progression of ligamentum flavum hypertrophy by up-regulating the expression levels of TGF-β1 and collagen type I. 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 Figures and Tables | References | Related Articles | Metrics

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Culture of fibroblast-like synoviocytes from rabbits with adjuvant arthritis Wang Li-ping, Liu Qin, Chen Fang, Chen Li-feng, Zhang Yi 2017, 21 (12):  1872-1876.  doi: 10.3969/j.issn.2095-4344.2017.12.012 Abstract ( 335 )   PDF (4406KB) ( 394 )   Save BACKGROUND: Currently, fibroblast-like synoviocytes from the synovial tissue of patients with rheumatoid arthritis or adjuvant arthritis rats are isolated and cultured mainly using enzyme digestion method, but tissue explant adherent method is rarely reported in primary culture. OBJECTIVE: To establish a culture method of fibroblast-like synoviocytes from adjuvant arthritis rabbits and to investigate cell biological characteristics. METHODS: Rabbit models of adjuvant arthritis were prepared by injected with adjuvant at multiple sites and times, the knee synovial tissues were obtained, and fibroblast-like synoviocytes were separated and cultured by the tissue explant adherent method. Furthermore, the cell morphology was observed. The cell proliferation was measured using cell counting kit-8 assay, and the expression of Vimentin was detected by immunofluorescence. RESULTS AND CONCLUSION: The primary cultured rabbit synoviocytes were long spindle fiber-like and coincided with the fibroblast-like synoviocytes. Moreover, cells were positive for Vimentin. In conclusion, fibroblast-like synoviocytes from rabbits with adjuvant arthritis are successfully isolated and cultured by the tissue explant method. 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 Figures and Tables | References | Related Articles | Metrics

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Establishment of an in vivo rabbit model of intervertebral disc degeneration under tensile load and its significance Xiao Liang, Xu Hong-guang, Shen Xiang 2017, 21 (12):  1877-1882.  doi: 10.3969/j.issn.2095-4344.2017.12.013 Abstract ( 340 )   PDF (5639KB) ( 681 )   Save BACKGROUND: Abnormal stress is an important factor causing intervertebral disc degeneration. To establish an ideal in vivo animal model of intervertebral disc degeneration is of great significance for in-depth study on the related pathogenesis. OBJECTIVE: To develop an in vivo rabbit intervertebral disc model and to investigate the relationship between continuous tensile load and intervertebral disc degeneration. METHODS: Twenty-five New Zealand white rabbits aged 6 months old were randomly divided into three groups: blank control (n=5), sham (n=10) and experimental (n=10) groups. The blank control group received no intervention, and the L4/5 segments were removed at the 1st day. The intervertebral disc assistor was used to fix the L4 and L5 vertebral bodies in the experimental and sham groups, the L4/5 segments in the experimental group were loaded 1 MPa axial tensile force, and the L4/5 segments in both two groups were then removed at 14 and 28 days. The changes of L4/5 intervetebral space height and surrounding bone substance were observed by X-ray examination, the morphological changes of the intervertebral disc were observed by hematoxylin-eosin staining, the cell survival was detected by nitro blue tetrazolium staining and mRNA expression levels of aggrecan, collagen type II and SOX9 in the intervertebral disc tissues were assessed by RT-PCR at each time point. RESULTS AND CONCLUSION: The radiological manifestations, histological changes, cell survival and mRNA expression levels of aggrecan, collagen type II and Sox9 showed no significant difference between the blank control and sham groups. Comparied with the blank control group, in the experimental group, the L4/5 intervertebral space was narrowed with time, the articular surface was coarse, and the upper and lower corpus vertebrae edge appeared to have lip-shaped hyperplasia; the intervertebral disc cells distributed irregularly; the nucleus pulposus was in dehydration and deflation, annulus fibrosus arranged irregularly, and the vacuoles in notochord cells tended to disappear; the expression levels of aggrecan, collagen type II and SOX9 were markedly downregulated. These findings suggest that the in vivo rabbit model of intervertebral disc is successfully established, in which continuous mechanical tensile load is further proved to directly cause intervertebral disc degeneration. 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 Figures and Tables | References | Related Articles | Metrics

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Morphology of the intervertebral disc in high fat combined with streptozotocin-induced type II diabetic rats Liu Guang-yuan, Dai Mu-wei, Tian Fa-ming, Yuan Lei-liang, Geng Lin-dan, Bei Ming-jian, Zhang Liu, Wang Wen-ya 2017, 21 (12):  1883-1888.  doi: 10.3969/j.issn.2095-4344.2017.12.014 Abstract ( 278 )   PDF (1488KB) ( 449 )   Save BACKGROUND: Rat models of diabetes mellitus type 2 (T2DM) are usually induced by the combination of high-fat diet and low-dose streptozotocin, but their effects on the intervertebral disc have not yet been reported. Endplate sclerosis is an important factor contributing to intervertebral disc degeneration. OBJECTIVE: To evaluate whether the rat T2DM model induced by high-fat diet combined with low-dose streptozotocin is suitable for the study of T2DM related intervertebral disc degeneration. METHODS: Thirty-two 3-month-old female Sprague-Dawley rats were divided into four groups (n=8 per group), including sham, bilateral variectomy, DM, and bilateral variectomy plus DM groups, followed by subjected to bilateral ovariectomy and/or high-fat diet combined with low-dose streptozotocin, respectively. The blood glucose level, body mass and glucose tolerance were determined. The bone mineral density of the lumbar spine was measured after 8-week streptozotocin treatment. The L5-6 segments were removed and cut through midst sagittal plane after decalcification, and then underwent Von Gieson staining and histological degeneration scoring, and the disc height and endplate thickness were measured. RESULTS AND CONCLUSION: Fasting blood glucose and random blood glucose levels in the DM and bilateral variectomy plus DM groups were significantly higher than those in the other two groups, and the insulin sensitivity in the DM and bilateral variectomy plus DM groups were significantly lower than that in the other groups (P < 0.05). L4-6 vertebral bone mineral density in the bilateral variectomy group was significantly lower than that in the sham group (P < 0.05); L5-6 vertebral bone mineral density in the DM and bilateral variectomy plus DM groups was significantly lower than that in the sham group (P < 0.05). L5-6 vertebral histological scores in the DM and bilateral variectomy plus DM groups were significantly higher than those in the other groups (P < 0.05). Similar with the bilateral variectomy group, there were chondrometaplasia and mucoid degeneration of nucleus pulposus cells in the bilateral variectomy plus DM group, and the histological scores were significantly higher than those in the sham and DM groups (P < 0.05). Compared with the sham group, the intervertebral disc height in the bilateral variectomy and bilateral variectomy plus DM groups was significantly decreased (P < 0.05). While, there was no significant difference in the endplate thickness among groups. These results indicate the combination of high-fat diet and low-dose streptozotocin-induced rat T2DM models possess diabetic characteristics, but the rat intervertebral disc tissues show no significant differences from the normal ones; therefore, this model may be unsuitable for the study on T2DM-related intervertebral disc degeneration. 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 Figures and Tables | References | Related Articles | Metrics

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Establishment of a three-dimensional model of rabbit lumbar vertebrae: its morphological characters He Qing1, Li Bing1, Deng Yan-qing2, Zhuo Xiang-long1, Jiang Yu1 2017, 21 (12):  1889-1893.  doi: 10.3969/j.issn.2095-4344.2017.12.015 Abstract ( 403 )   PDF (1071KB) ( 278 )   Save BACKGROUND: Morphological characteristics of the rabbit lumbar vertebrae are of great significance for animal experiments about lumbar spine, but there is a lack of study on the lumbar morphology in rabbits. OBJECTIVE: To establish a three-dimensional model of rabbit lumbar vertebrae, and to observe its morphological characteristics, thereby providing anatomical basis for the animal experiments about lumbar spine.  METHODS: Twenty New Zealand rabbits were subjected to 64-row multi-detector CT (Siemens) scan, and the three-dimensional models were constructed based on the CT data from 140 lumbar vertebrae. The morphological parameters of L1-7 were measured, and the internal and external structures of lumbar vertebrae and changes of different segments were observed. RESULTS AND CONCLUSION: (1) Changes of the morphological parameters of each lumbar vertebra: the L1-7 height first increased and then decreased, and the shortest height was in the L7, which was (1.12±0.18) cm. L1 vertebral body exhibited the shortest transverse diameter, (1.03±0.15) cm, and the longitudinal diameter did not differ significantly among vertebrae. The transverse and longitudinal diameters of L1-6 spinal canals were on a rise, but decreased abruptly at L7. The pedicle width of L1-7 increased gradually; the pedicle height of L1-5 increased gradually, and then began to decrease markedly, and the shortest at L7, (0.58±0.11) cm. The width and length of L1-6 spinous process increased steadily, but decreased at L7. The length of L1-3 transverse process increased gradually, tended to be steady at L3-6, and increased notably at L6-7; there were significant differences in the transverse process angle at L1-6, but the angle increased markedly to (58±2)° at L7, (2) These results indicate that the internal and external structures of lumbar vertebrae and changes of different segments are obtained through morphological measurement of lumbar vertebrae in New Zealand white rabbit, which provides a detailed anatomical data for relative animal experiments. 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 Figures and Tables | References | Related Articles | Metrics

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Establishment and analysis of a rat model of chronic renal allograft dysfunction complicated with insulin resistance Lv Dao-yuan1, 2, Zhou Qin1, 2, Xia Yue1, 2, You Xu1, 2, 3, Zhao Zhi-hong4, Li Yong-qiang1, 2, Zou He-qun1, 2 2017, 21 (12):  1894-1902.  doi: 10.3969/j.issn.2095-4344.2017.12.016 Abstract ( 293 )   PDF (9216KB) ( 424 )   Save BACKGROUND: The main cause of chronic renal allograft dysfunction (CRAD) remains unclear until now, and insulin resistance (IR) may be a cause of CRAD. However, there is a lack of an available animal model for research on their association. OBJECTIVE: To establish a rat model of CRAD complicated with IR, and to investigate the effect of IR on function, urinary protein and pathologic changes of the renal allograft. METHODS: The high-fat diet was administrated to a F344-to-Lewis rat model of CRAD to induce IR. There were three groups including CRAD+IR, CRAD and control (uninephrectomy) groups, and the serum insulin, glucose, creatinine, urea nitrogen and 24-hour urinary protein levels in each group were measured and insulin resistance index was calculated at 8, 12, and 16 weeks after transplantation. Meanwhile, oral glucose tolerance and insulin tolerance tests were performed and the areas under glucose-time curve were calculated. The status and stability of CRAD+IR models were assessed based on all above indicators at each time point. The serum creatinine, urea nitrogen and 24-hour urinary protein levels were compared between CRAD+IR and CRAD groups. Pathological evaluation was conducted at 16 weeks based on the total Banff scores. RESULTS AND CONCLUSION: The insulin resistance index, and area under glucose-time curve of the oral glucose tolerance and insulin tolerance tests in the CRAD+IR group were higher than those in the CRAD and control groups, and all above indexes showed no significant changes with time. The serum creatinine, urea nitrogen and 24-hour urinary protein levels in the CRAD+IR group were lower than those in the CRAD group, and showed no obvious changes with time. Slightly thickened basement membrane of glomeruli and tubules, interstitial fibrosis and mild arterial intimal hyperplasia were observed both in the CRAD+IR and CRAD groups, but the pathological degree did not differ notably between the two groups. These results indicate that the stable CRAD+IR models can be established via feeding CRAD rats with high-fat diet for 8 weeks. IR can cause glomerular hyperfiltration and albumin overload-induced tubular injury which may potentially affect the function and urinary protein level of the renal allograft. 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 Figures and Tables | References | Related Articles | Metrics

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Construction of a high efficient pancreatic duodenal homeobox 1/insulin dual-reporter vector and its preliminary application Ye Ling-ling1, Li Shi-chong1, Sun Hai-yan2, Lan San-chun3, Chen Zhao-lie1, Wang Qi-wei1 2017, 21 (12):  1903-1908.  doi: 10.3969/j.issn.2095-4344.2017.12.017 Abstract ( 279 )   PDF (4228KB) ( 545 )   Save BACKGROUND: Islet beta cell replacement therapy is one of the most promising approaches for treating type 1 diabetes mellitus. However, its large scale application is hampered by a shortage of islet beta cells for transplantation. Pluripotent stem cells are one of ideal seed cells for islet beta cell replacement therapy, but pancreatic beta-cell differentiation is time-consuming and labor-intensive. OBJECTIVE: To construct a high efficient pancreatic and duodenal homeobox 1 (Pdx1)/insulin dual-reporter vector and to monitor the key genes expression during pancreatic beta-cell differentiation from pluripotent stem cells. METHODS: In order to construct a high efficient Pdx1/insulin dual-reporter vector, puromycin resistance gene was firstly introduced into pTiger vector, and then the original 410 bp mouse Ins1 promoter of the vector was replaced by 646 bp mouse Ins1 promoter. Finally, the dual-reporter vector was transduced into INS-1 and human induced pluripotent stem cells to testify its function. RESULTS AND CONCLUSION: The high efficient Pdx1/insulin dual-reporter vector was constructed successfully. The vector successfully acquired puromycin resistance gene and high gene expression efficacy of insulin in INS-1 cells. The specific gene expression pattern of Pdx1/insulin was first found in INS-1 cells. To conclude, the real-time monitoring function of Pdx1/insulin expression is preliminarily confirmed during pancreatic beta-cell differentiation. 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 Figures and Tables | References | Related Articles | Metrics

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Effect of metformin on the insulin resistance in a rat model of type 2 diabetes mellitus Qin Feng1, Li Chun-liang2, Zhang Hui-li1 2017, 21 (12):  1909-1914.  doi: 10.3969/j.issn.2095-4344.2017.12.018 Abstract ( 278 )   PDF (1951KB) ( 402 )   Save BACKGROUND: More than 10 kinds of lipid metabolism-related genes have been found, and endocrine dysfunction mediated by these genes is an important pathological basis for the occurrence and development of insulin resistance. OBJECTIVE: To investigate the effects of metformin on serum lipid profiles and the expression levels of various  genes associated with insulin resistance, as well as the histopathological changes of the liver and pancreas in rats with type 2 diabetes melitus. METHODS: A type 2 diabetes mellitus rat model was established by feeding a high-fat diet to the rats for 4 weeks, combined with the intraperitoneal injection of streptozotocin (35 mg/kg). In the meanwhile, metformin was administered orally (400 mg/kg•d) (model group) or nothing (control group). Semi-quantitative RT-PCR, histopathological and biochemical examinations were then performed. RESULTS AND CONCLUSION: Metformin improved the symptoms of insulin resistance by normalizing the serum lipid profiles in the diabetic rats. Furthermore, metformin upregulated the expression levels of insulin receptors and genes associated with lipid metabolism, including acyl-CoA oxidase, carnitine palmitoyl transferase-1 and peroxisome proliferator activated receptor-α. In addition, metformin downregulated the expression levels of fetuin-A and retinol binding protein-4, and improved the expression of perilipin that had been reduced in the type 2 diabetes mellitus rats. Metformin was shown to induce positive signaling for insulin and the regeneration of pancreatic β cells in the pancreas. These results suggest that metformin ameliorates the insulin resistance induced by type 2 diabetes mellitus via regulating the expression levels of fetuin-A, retinol binding protein-4, perilipin and various genes associated with lipid metabolism. 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 Figures and Tables | References | Related Articles | Metrics

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Optimal acting time of cytarabine in primary culture of rat cortical neurons Guan Hong1, Pan Xue-feng2, 3, Liu Hao-kun4, Liu Xiao-qing1, Zhang Li-na2, Wang Shao-yi2, Dong Xiao-dong2, Niu Si-yun2 2017, 21 (12):  1915-1920.  doi: 10.3969/j.issn.2095-4344.2017.12.019 Abstract ( 493 )   PDF (1249KB) ( 438 )   Save BACKGROUND: Toxic cytarabine is often used to prepare highly purified neurons in experimental studies addressing central nervous system diseases. However, the intervention time of cytarabine is little reported. OBJECTIVE: To determine the optimal intervention time of cytarabine(final concentration 10 μmol/L) in primary culture of rat cortical neurons. METHODS: Rat primary cortical neurons were cultured in Neurobasal+B27 medium, and 10 μmol/L cytarabine were added at 12, 24, 36 and 48 hours after culture, respectively. Half of the medium were changed every 48 hours. The morphology of neurons was observed under inverted microscope at 7 days. The purity and differentiation of neurons maturity were identified by immunocytochemistry method of neuron specific enolization enzyme staining. Morphometric analysis for all neuron-specific enolase positive cells was performed by Multifunction Computer Image Analysis System. RESULTS AND CONCLUSION: After addition of cytarabine at 24 hours of culture, the purity of neurons was more than 90%, well-differentiated cortical neurons accounted for 89.00%, and the area of neuronal body was the largest with the longest synapses. There were more neuron cells with transparent cytoplasm, and large nucleus. The cell body had good refraction and strong stereo sense. The neurons with 3-4 synapses and 3-4 bifurcates formed a good network structure. These results illustrate that although it will be beneficial for the purity of neurons to add cytarabine early in neuron culture process, it will make obvious effect on neuronal differentiation. The highly purified and well-grown cerebral cortical neurons will be obtained after cultured in neurolbasal medium, which cytarabine is added to at 24 hours of culture. 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 Figures and Tables | References | Related Articles | Metrics

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Application of autologous hard palate mucosa in the eyelid reconstruction Shi Jun-hu1, Liu Shan1, Bai Ping1, Chu Miao2 2017, 21 (12):  1921-1925.  doi: 10.3969/j.issn.2095-4344.2017.12.020 Abstract ( 303 )   PDF (992KB) ( 471 )   Save BACKGROUND: Autologous hard palate mucosa has been validated to be a substitute of the tarsal conjunctival. OBJECTIVE: To investigate the treatment outcomes of autologous hard palate mucosa applied in the eyelid reconstruction. METHODS: 180 patients (180 eyes) with eyelid defects aged 10-63 years old were recruited, including 122 males and 58 females, and subjected to the transplantation of autologous hard palate mucosa. In the same time, flaps were designed and transferred into the defected eyelid soft tissues. All patients were followed up for over 3 months to observe the treatment outcomes. RESULTS AND CONCLUSION: Three months after surgery, viscous secretions were found in the implanted region and conjunctival sac, which were relieved and eliminated by usage of antibiotic drops. All implanted hard palate mucosa survived within 1 year, neither infections nor falling off occurred, and blood vessels growth into the implant-tissue conjugation that inosculated with the host vessels presenting no rejection. The implants revealed different degrees of shrink in 132 patients, but the eye function was normal. During the hospitalization, some uncomfortable feeling such as eye pain had been relieved, and the wound showed no infections and healed well. During the follow-up, there were eight patients with slight lower eyelid ectropion and three with hypophasis. The length or height difference of both eyelids was within 2 mm, except those who lost eye balls. These results suggest that autologous hard palate mucosa achieve satisfactory outcomes in the eyelid reconstruction and possesses good histocompatibility. 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 Figures and Tables | References | Related Articles | Metrics

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Application and progress of co-culture systems in cartilage tissue engineering Zhang Yu1, Liu Shu-yun1, Guo Wei-min1, Hao Chun-xiang2, Wang Ming-jie1, Lu Liang3, Lu Shi-bi1, Guo Quan-yi1 2017, 21 (12):  1926-1932.  doi: 10.3969/j.issn.2095-4344.2017.12.021 Abstract ( 350 )   PDF (1044KB) ( 384 )   Save BACKGROUND: Accumulative evidence supports that co-culture technology can be applied to construct the tissue-engineered cartilage with excellent biological characters. OBJECTIVE: To elaborate the co-culture concept and conclude and analyze seed cell sources, cell mixed ratio, spatially-defined co-culture models and biomaterials in co-culture systems to conclude and analyze the biological characters of tissue-engineered cartilage, and to prospect progression of co-culture systems in cartilage tissue engineering. METHODS: The first author retrieved the databases of PubMed, Web of Science, and CNKI for relative papers published from January 1976 to May 2016 using the keywords of “co-culture, co-culture systems; articular cartilage, chondrocytes, mesenchymal stem cells; tissue engineering, articular cartilage tissue engineering” in English and Chinese, respectively. Finally 60 literatures were included in result analysis, including 1 Chinese and 59 English articles. RESULTS AND CONCLUSION: Co-culture technology emphasizes the role of microenvironment in terms of various physical, chemical and biological factors in the cell processing. In cartilage tissue engineering, co-culture systems contribute to maintain the viability and natural cell phenotype of chondrocytes and induce cartilage differentiation of mesenchymal stem cells. In addition, co-culture technology provides a novel way for cartilage tissue engineering to overcome the shortage of chondrocytes and repair injury to the cartilage-subchondral bone. However, the mechanisms of cell-cell interaction in co-culture systems still need to be explored in depth, so as to optimize the co-culturing conditions and construct perfect tissue-engineered cartilage. 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 Figures and Tables | References | Related Articles | Metrics

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Clinical manifestations of osteoarthritis and the role of silent information regulation 1 in the pathogenesis of osteoarthritis Lei Ming1, Yu Fei2, Xiao De-ming1 2017, 21 (12):  1933-1939.  doi: 10.3969/j.issn.2095-4344.2017.12.022 Abstract ( 352 )   PDF (1115KB) ( 467 )   Save BACKGROUND: Functional changes of some functional genes have been showed to trigger osteoarthritis, among which, age may be a critical one. OBJECTIVE: To summarize the clinical manifestations of osteoarthritis and the research process of silent information regulation 1 in the occurrence and development of osteoarthritis. METHODS: A computer-based retrieval was performed in the databases of CNKI, PubMed, SpringerLink, and Elsevier Science Direct using the keywords of “osteoarthritis, silent information regulation 1 and cartilage” in Chinese and English, respectively. Finally 83 eligible literatures were enrolled for analysis. RESULTS AND CONCLUSION: Deacetylation modification acts on the occurrence and development of osteoarthritis by regulating the expression and biological function of various cytokines. Silent information regulation 1 cannot only regulate body metabolism, inhibit cell apoptosis, repair DNA injury, delay senescence and resist to stress, but also play important roles in the epigenetic modification, gene silencing and signal transduction. Therefore, silent information regulation 1 is involved in the age-related diseases, such as osteoarthritis, osteoporosis, diabetes and Alzheimer’s disease. Furthermore, silent information regulation 1 is likely to be an important target of osteoarthritis drugs, and even reverses mechanical stress-induced cartilage injury in the early stage of osteoarthritis. 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 Figures and Tables | References | Related Articles | Metrics

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Ghrelin participates in bone metabolism: the newest research progress Chen Hong1, Bi Ran-ran2, Chen Liang1, Zhu Qiong1 2017, 21 (12):  1940-1945.  doi: 10.3969/j.issn.2095-4344.2017.12.023 Abstract ( 373 )   PDF (1112KB) ( 310 )   Save BACKGROUND: Ghrelin is a newly discovered brain-gut peptide from the stomach of human and rats. As an endogenous ligand for growth hormone secretagogue receptor (GHSR), ghrelin can notably stimulate the release of growth hormone. Although GHSR is expressed in many peripheral tissues, little is known about the influence of ghrelin on bone metabolism and GHSR expression in bone tissue. OBJECTIVE: To review the research progress of ghrelin in bone metabolism. METHODS: The first author retrieved CNKI, WanFang, PubMed, and Springerlink databases with the keywords of “Ghrelin, bone metabolism” in Chinese and English, respectively. The studies regarding ghrelin and its involvement in bone metabolism were included, and repetitive ones were excluded. A total of 53 eligible literatures were selected through skimming abstracts. RESULTS AND CONCLUSION: Ghrelin is a peptide composed of 28 amino acids discovered in gastric endocrine cells and hypothalamic arcuate nucleus in mice and human, which makes a great effect on digestive, nervous, immune and endocrine systems, and also plays a role in hormone secretion, glucose metabolism, immunity, cell proliferation, and inflammation. Serum ghrelin makes a certain influence on bone growth and development, and promotes the differentiation and proliferation of osteoblasts, and inhibits its apoptosis. Additionally, ghrelin suppresses the early osteogenic differentiation of C3H10T1/2 cells by upregulating the expression of Runx2 protein, and attenuates adipogenic differentiation by downregulating PPARγ2 expression, thus inducing osteogenic differentiation. However, few studies have addressed the expression of GHSR in bone tissue. 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 Figures and Tables | References | Related Articles | Metrics

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Effects of mechanical factors on autophagy  Gui Jin-peng, Jiang Jing-yi, Zhang Ling-yu, Na Jing, Fan Yu-bo, Zheng Li-sha 2017, 21 (12):  1946-1952.  doi: 10.3969/j.issn.2095-4344.2017.12.024 Abstract ( 433 )   PDF (1343KB) ( 320 )   Save BACKGROUND: Numerous diseases like cardiac hypertrophy and intervertebral disc degeneration are known to be implicated in the changes of mechanical stress acting on surrounding tissues or cells, and autophagy contributes to the pathogenesis of these diseases. OBJECTIVE: To review the effects of mechanical factors on autophagy in different tissues and the underlying molecular mechanisms, thereby providing references for the research of autophagy and the prevention and treatment of related diseases. METHODS: A search of Web of Science and PubMed databases was performed for the literatures addressing the effects of mechanical factors on autophagy using the English keywords of “autophagy, mechanical” and the articles were summarized systematically. Finally, 52 literatures were enrolled according to the inclusion and exclusion criteria. RESULTS AND CONCLUSION: Mechanical factors make great effects on autophagy of various cells, such as myocardial cells, endothelial cells, chondrocytes and skeletal muscle cells. Autophagy is a self-protective reaction, and the mechanical stress of physiological conditions induces autophagy to maintain cell homeostasis, normal function and survival. The mechanism of autophagy induced by mechanical stress may involve PI3K-AKT-mTOR, oxygen free radical, AKT-FoxO and other pathways, and the definite mechanism needs to be further studied. 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 Figures and Tables | References | Related Articles | Metrics

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Safety management for tissue engineering laboratories in universities Wang Ling1, 2, Luo Li1, Xu Ming-en1, 2 2017, 21 (12):  1953-1958.  doi: 10.3969/j.issn.2095-4344.2017.12.025 Abstract ( 290 )   PDF (955KB) ( 377 )   Save BACKGROUND: Tissue engineering is an interdisciplinary field and its experimental safety covers a wide scope along with the increase in accidents of tissue engineering laboratories, the safety management in tissue engineering laboratories in universities has become an important public health problem. OBJECTIVE: To enhance the safety management strategies for tissue engineering laboratories in chinese universities and improve the safety management system. METHODS: PubMed, Elseveir, WanFang and CNKI databases were retrieved for the literatures concerning  the biosafety, safety management of hazardous chemicals, waste, medical information, instrument operation and human body mechanics in tissue engineering laboratories, and then the status of laboratory management was analyzed. RESULTS AND CONCLUSION: The status of safety management in tissue engineering laboratories in China is analyzed based on the technique characters and regulations on safety management. Management strategies for laboratory safety have been proposed according to the differences in relevant laboratory safety management between China and the US, such as regular training on laboratory safety, improving the safety consciousness of experimental staff, constructing the strict system access to laboratory, improving safety facilities, standardizing the experimental safety manual and appointing specialized safety management committee. Comprehensive and systematical regulations on laboratory safety management will ensure the smooth running of tissue engineering laboratories in universities, thus accelerating the development of tissue engineering. 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 Figures and Tables | References | Related Articles | Metrics

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Architecture of network medicine and its research progress Li Ying1, Huang Wen-cong2 2017, 21 (12):  1959-1968.  doi: 10.3969/j.issn.2095-4344.2017.12.026 Abstract ( 349 )   PDF (1343KB) ( 472 )   Save BACKGROUND: Network medical research contains various widely applicable networks with different structures, and construction methods. These networks are not organized in architecture yet, and there is a lack of overview about its application and research progress. OBJECTIVE: To organize various medical networks to a hierarchical architecture, and summarize their research progress. METHODS: A computer-based search of CNKI and PubMed databases was performed for articles about the network medicine published between 1999 and 2016 using the keywords of “disease network, protein interaction network, metabolic network, gene regulatory” in English and Chinese, respectively. Then the main research contents, module property, construction methods, maturity and application progress of network medicine were discussed to predict the research direction. RESULTS AND CONCLUSION: The architecture of network medicine consists of molecular interaction network layer (interactome), disease network layer (diseasome) and drug network layer (drugome). In the interactome, protein-protein interaction network, metabolic network and gene regulatory network are in the order of organisms → mammals → human, and respective networks of human have been mapped but not mature; and new networks are being built. In diseasome, many disease networks have been constructed using a single element to cluster; and cluster by new element or combined elements can be considered in the future. Given this the current pharmacology is based on protein, there are few networks in the drugome the elements that constitute the drugome are only drug and protein. 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 Figures and Tables | References | Related Articles | Metrics