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08 October 2018, Volume 22 Issue 28 Previous Issue    Next Issue

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Lentivirus-mediated Runx2 gene transfection into MC3T3-E1 cells Qin Han, Gong Yong-qing, Xu Hong-zhi 2018, 22 (28):  4429-4433.  doi: 10.3969/j.issn.2095-4344.0951 Abstract ( 264 )   PDF (1295KB) ( 122 )   Save BACKGROUND: Heterozygous mutation, gene insertion and gene deletion of Runt-related transcription factor 2 (Runx2) have been found to be important causes of cleidocranial dysostosis. However, there is a lack of studies on the changes of osteoblasts after Runx2 silencing and how to perform gene silencing. OBJECTIVE: To construct the lentiviral vector targeting Runx2 gene in mouse osteoblast-like cells (MC3T3-E1), so as to provide experimental basis for the research of cleidocranial dysostosis. METHODS: According to the preliminary experimental results, the lentiviral vector was infected by MOI=40 in the ENi.S medium containing  5 mg/L Polybrene for 10 hours, and divided into four groups according to the viral number: group NC: pFU-GW-016PSC53349-1; group KD1: LVpFU-GW-016PSC60107-1; group KD2: LVpFU-GW-016PSC60108-1 and group KD3: LVpFU-GW-016PSC60109-1, then replaced with conventional medium at 16 hours after infection. The target gene expression was detected by Celigo® Image Cytometer at 72 hours after transfection. The two step method of real-time PCR was utilized to detect the silencing effect of Runx2 gene. RESULTS AND CONCLUSION: At 72 hours after transfection, there was no green fluorescence in the group NC; a small amount of fluorescent expression in the group KD1; enhanced fluorescence expression in the group KD2 and obvious green fluorescence in the group KD3. Real-time PCR curves showed the better silencing effect of Runx2 gene in the group KD3. In summary, we successfully constructed the lentiviral vector targeting Runx2 gene into MC3T3-E1 cells, screened the virus inhibiting Runx2 gene, and clarified the action time and related measurement. 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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Polymorphisms of interleukin-23 receptor gene and susceptibility of bone and joint tuberculosis in Guangxi Zhuang populations Wu Zhao-yuan1, Xie Ke-gong2, Lu Lu2, Lu Xian-zhe2, Huang Ke2, Liao Lin-bo1, Tang Yu-jin2 2018, 22 (28):  4434-4439.  doi: 10.3969/j.issn.2095-4344.0840 Abstract ( 241 )   PDF (1090KB) ( 102 )   Save BACKGROUND: The incidence of tuberculosis is high in populations from Guangxi Zhuang Autonomous Region. Tuberculosis has been shown to have susceptibility gene. OBJECTIVE: To analyze the relationship between interleukin-23 receptor (IL-23R) gene polymorphism and susceptibility to bone and joint tuberculosis in Guangxi Zhuang populations. METHODS: Totally 125 patients with bone and joint tuberculosis admitted in the Affiliated Hospital of Youjiang Medical University for Nationalities and 150 healthy subjects were selected. The gene polymorphism of IL-23R (rs10489628G/A, rs10489629T/C, rs10889675A/C, rs10889677C/T, rs7530511C/A) was analyzed by PCR amplification technique and DNA sequencing method. The genotypes and frequency distribution of alleles were compared between the tuberculosis and control groups. The relationship between IL-23R gene polymorphism and the susceptibility to bone and joint tuberculersis was discussed. RESULTS AND CONCLUSION: The frequency of C allele of IL-23R rs10489629 T/C in the tuberculosis group was 29.2%, which was significantly higher than that in the control group (21.3%) (P < 0.05). The frequency of A allele of IL-23R rs10889675 C/A in the tuberculosis group was 26.0%, which was significantly higher than that in the control group (18.0%). The frequency of CA genotype of IL-23R rs10889675 C/A in the tuberculosis group was 37.6%, which was significantly higher than that in the control group (25.3%) (P < 0.05). The frequency of GTCAC haplotype in the bone and joint tuberculosis group was 69.6% which was lower than that in the control group (77.3%). The frequency of ACACC haplotype in the bone and joint tuberculosis group was 26.0% which was higher than that in control group (17.7%). In summary, the gene polymorphism of IL-23R rs10489629T/C and rs10889675C/A may be related to the susceptibility of bone and joint tuberculosis in Guangxi Zhuang populations. 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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Combination of bone morphogenetic protein 2 and mutant hypoxia-inducible factor 1α alpha repairs steroid-induced avascular necrosis of the femoral head Hu Liang1, Wang Jun-hai1, Wang Zhi-lie1, Xie Jin-yuan1, Chen Deng1, Ding Fan2 2018, 22 (28):  4440-4446.  doi: 10.3969/j.issn.2095-4344.0193 Abstract ( 312 )   PDF (6039KB) ( 218 )   Save BACKGROUND: Bone morphogenetic protein 2 (BMP-2) is the unique growth factor that independently induces osteogenesis, but its promoting angiogenesis is weak, and cannot produce sufficient capillaries in the newly-born bones, thereafter, other auxilliary factors are necessary. Hypoxia-inducible factor 1α (HIF-1 α) plays an important role in promoting angiogenesis. OBJECTIVE: To study the repair capacity of the adenovirus mediated BMP-2 combined with mutant HIF1α (Ad-BMP-2-IRES-HIF-1αmu) transfecting bone marrow mesenchymal stem cells (BMSCs) in steroid-induced avascular necrosis of the femoral head. METHODS: Ad-BMP-2-IRES-HIF-1αmu was transfected to BMSCs, the alkaline phosphatase activity and calcium nodules alizarin red staining were conducted to detect the osteogenic ability. The rabbit model of steroid-induced avascular necrosis of the femoral head was established, and then divided into three groups: The cells were divided into three groups: Ad-BMP-2-IRES-HIF-1αmu transfected BMSCs were transplanted into group A, and untransfected BMSCs were transplanted into group B, and only the cell culture medium was added in the group C. At 8 weeks after transplantation, the expression levels of BMP-2 and HIF-1α were detected by western blot assay; the repair of avascular necrosis of the femoral head was observed by hematoxylin-eosin staining; the angiogenesis of the necrotic region was observed by toner perfusion transplantation vascular morphology.  RESULTS AND CONCLUSION: The alkaline phosphatase activity in the transfected BMSCs and number of calcium nodules were higher than those in the untransfected BMSCs, indicating that Ad-BMP-2-IRES-HIF-1αmu could promote the osteogenic differentiation of BMSCs. The expression levels of BMP-2 and HIF-1α in the group A were significantly higher than those in the groups B and C (P < 0.05). Compared with groups B and C, obvious osteogenesis was observed in group A, the percentage of empty lacunae was significantly decreased, and trabecular bone volume fraction was significantly increased (P < 0.05). Ink perfusion results showed that compared with the groups B and C, there was a vascularization in the group A, and there was a structure of bone lacuna, an increase in blood vessel diameter, a clear connection between blood vessels, and rich and effective blood vessels in the defect area (P < 0.05). The blood vessel area in the three groups was 114.22 ± 3.78, 63.78 ± 2.61 and 21.39 ± 3.52, respectively (P < 0.05). In conclusion, BMP-2 combined with mutant HIF1α can promote the repair of steroid-induced avascular necrosis of the femoral head. 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 | Related Articles | Metrics

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Puerarin promotes osteogenesis of osteoblasts compounded with beta-tertiary calcium phosphate Chang Zai-ping1, Yang Hai-long2, Zhang Ming-yu3 2018, 22 (28):  4447-4451.  doi: 10.3969/j.issn.2095-4344.0388 Abstract ( 333 )   PDF (4206KB) ( 131 )   Save BACKGROUND: Puerarin is the major bioactive ingredient isolated from the root of the Pueraria lobata (Willd.) Ohwi, which is a kind of isoflavones. Isoflavones has been shown to promote the proliferation and differentiation of osteoblasts. OBJECTIVE: To observe the effect of puerarin at different concentrations on the osteogenic activity of β-tertiary calcium phosphate (β-TCP) complex, and to investigate the effect of puerarin on osteogenesis in vivo. METHODS: Osteoblasts isolated from calvarial of newborn rabbits were cultured in vitro with different concentrations of puerarin. The alkaline phosphotase activity and mineral node formation were determined by alkaline phosphatase kit and alizarin red staining. The osteoblasts cultured with the optimal concentration of puerarin (group A) or without puerarin (group B) were seeded on β-TCP, respectively. Osteoblast activity was determined using MTT assay. Scaffold complexes in each group were implanted into the rabbit dorsal muscles. Osteogenesis was analyzed by histological analysis. RESULTS AND CONCLUSION: The alkaline phosphatase activity and mineral nodules were significantly increased in osteoblasts cultured with puerarin, and 1×10-6 mol/L puerarin could significantly incresase increase the osteoblast activity in the scaffold. After the β-TCP composite scaffolds cultured with 1×10-6 mol/L puerarin were implanted into the rabbit dorsal muscles, the newly formed bone was significantly increased. In summary, puerarin enhances the osteogenic effect of β-TCP osteoblast complex in vivo, which can be used to promote osteogenesis. 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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Correlation of bone mineral density with sports ability Chen Zhen-yong 2018, 22 (28):  4452-4456.  doi: 10.3969/j.issn.2095-4344.0272 Abstract ( 696 )   PDF (4050KB) ( 90 )   Save BACKGROUND: Sports effects on bone mineral density (BMD) are found to be related to sport events, intensity and duration time. OBJECTIVE: To understand the relationship between BMD, especially the radius BMD and sports ability. METHODS: Totally 173 subjects were recruited, including 93 basketball athletes (21 elite athletes, 32 first-class athletes and 40 second-class athletes), and 80 sophomores. The BMD of bilateral radius, hip bone, femur, calcaneus, and lumbar vertebrae was measured by Prodigy Advance dual-energy X-ray absorptiometry. The independent sample t test, one-way ANOVA and Pearson correlation analysis were conducted using SPSS 20.0 software. RESULTS AND CONCLUSION: Except for the BMD of lumbar vertebrae, all measurements were significantly different between basketball players and college students (P < 0.05), especially the radial BMD (P < 0.01). Except for the BMD of left radius in male athletes and bilateral radius in female athletes of the elite and first-class groups, all measurements showed significant differences among athletes (P < 0.05). The correlation analysis between basketball ability and radical BMD was stronger in males than in females on the right side than on the left side. Therefore, long-term basketball exercise has a positive impact on appendicular BMD. The radical BMD is positively related to the basketball ability, so it can provide a theoretical basis for the optimization of the index selection of basketball players. 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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Relationship of membrane type-1 matrix metalloproteinase expression and chondrocyte migration in adult minced articular cartilage Dai Zhu1, Peng Jia-bin1, 2, Liao Ying1, Fu De-hong1, Lei Yun-liang1, Li Zhou1 2018, 22 (28):  4457-4462.  doi: 10.3969/j.issn.2095-4344.0837 Abstract ( 275 )   PDF (5094KB) ( 247 )   Save BACKGROUND: Minced articular cartilage has been shown to be helpful for cartilage repair, but the underlying mechanism remains unclear. OBJECTIVE: To observe the association of expression level of membrane type-1 matrix metalloproteinase (MT1-MMP) with chondrocyte migration, and to further research the underlying mechanism. METHODS: Massive cartilage (5 mm of diameter, and 2 mm of thickness) and articular cartilage fragments (about 1 mm3) were obtained from 78 adult rabbit keen joints under sterile condition, and then planted into fibrin glue-gelatin sponge composite scaffolds. Group A: cartilage fragments cultured in vitro with normal culture medium; group B: monoblock cartilage cultured in vitro with normal culture medium; group C: cartilage fragments cultured in vitro with culture medium containing inhibitor of MT1-MMP (NSC402050). The specimens were taken out respectively from each group after culture for 2, 4 and 6 weeks, for observing the chondrocyte migration under confocal laser scanning microscope, hematoxylin-eosin staining and histological semi-quantitative scoring. The expression level of MT1-MMP was detected by immunohistochemistry in the groups A and B, and the percentage of cells positive for MT1-MMP was calculated. RESULTS AND CONCLUSION: Laser confocal microscope results showed that chondrocytes gathered at the margin of tissue after 2 and 4 weeks of in vitro culture in the groups A and B, especially the group A, and at the same time, there were numerous chondrocytes. Group C showed no chondrocyte gathered at the margin of tissue and there was no cell release. Semi-quantitative score and the percentage of cells positive for MT1-MMP in the groups A and B were increased with time in vitro, and the scores and MT1-MMP positive cells in the group A were higher than those in the group B. The semi-quantitative score in the group C was closed to 0. These results suggest that the minced articular cartilage can promote cartilage repair via up-regulating the expression level of MT1-MMP and increasing the migration of chondrocytes. 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 | Related Articles | Metrics

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Stamping tensile test in rabbits: differences in mechanical parameters between veins and arteries Zhang Yu-hao1, 2, Niu Pei1, Zhang Zhi-min2, Niu Xiao-long1, Shen Wen-zeng1, Zhou Yu-juan1, Liu Fu-lin2 2018, 22 (28):  4463-4468.  doi: 10.3969/j.issn.2095-4344.0389 Abstract ( 405 )   PDF (4866KB) ( 125 )   Save BACKGROUND: There are few studies on the graft patency and biomechanics following coronary artery bypass graft surgery. OBJECTIVE: To analyze the differences in mechanical parameters between veins and arteries. METHODS: Twenty rabbits were randomized into control and experimental groups (n=10 per group). One side of the rabbit external jugular veins in the experimental group and the rabbit carotid arteries in the control group were removed and stored into the preservation solution (no Ca+ solution) for stamping tensile test. The external diameter of veins and arteries under different pressures (0-26.6 kPa), active state (high K+ solution) and passive state was measured. The vascular morphology was observed by hematoxylin-eosin staining. The stamping tensile test results were analyzed. RESULTS AND CONCLUSION: Under passive or active state, the vascular outer diameter (inner diameter)-pressure curve, vascular wall thickness-pressure curve and circumferential stretch ratio-pressure curve tended to be stable. The elasticity of the veins was significantly inferior to that of the arteries. After stamping, the outer diameter of veins began to increase, and was twice of the arteries finally. Stereomicroscopy observed that the venous wall was thinner. With the internal pressure increasing, the arterial and venous outer diameter tended to be stable at 13.3 and 5.32 kPa, respectively. The vessel cross-section results showed that the thickness of the veins was significantly smaller than that of the arteries. To conclude, with the intraluminal pressure increasing, venous elasticity peaked earlier, and its compliance was poor, so it was vulnerable to high pressure, which is in accordance with the thinner elastic layer of venous wall. 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 different volume fractions of platelet-rich plasma on the proliferation and migration of vascular endothelial cells Zhang Ming-yu, Ren Bo, Yang Zhen, Zhang Xian, Zheng Jiang 2018, 22 (28):  4469-4474.  doi: 10.3969/j.issn.2095-4344.0331 Abstract ( 354 )   PDF (4506KB) ( 129 )   Save BACKGROUND: Platelet-rich plasma (PRP) plays an important role in promoting angiogenesis of vascular endothelial cells. Different volume fractions of PRP have definite influence on vascular endothelial cells, but it is not clear PRP at which volume fraction is optimal. OBJECTIVE: To investigate the effect of PRP at different volume fractions on the proliferation and migration of vascular endothelial cells in vitro. METHODS: Hearts of Sprague-Dawley rats were mechanicaIly minced and enzymatically digested with collagenase. Vascular endothelial cells were counted and cultured in the DMEM culture medium containing various volume fractions of autologous PRP (10%, 20%, 50%) respectively. The cell morphology was observed under an inverted microscope, and cellular proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Afterwards, scratch test was utilized to detect the cell migration ability and the optimal volume fraction was determined. RESULTS AND CONCLUSION: After addition of PRP, the well-grown endothelial cells presented with regular shape, changed into polygon-like and became large with time. Different volume fractions of PRP in particular 20% PRP cultured for 72 hours could promote cell proliferation. According to scratch test, 20% and 50% PRP significantly promoted the cell migration, especially 20% PRP. These results indicate that PRP can promote the proliferation and migration of vascular endothelial cells in vitro, and 20% is the optimal volume fraction. 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 baicalin on human periodontal ligament fibroblasts and the underlying molecular mechanism Li Li-hua1, Jia Ai-min2, Yang Ming-hui2, Liu Zhen1, Wang Li-heng2, Jiang Hong2, Qiu Ya2 2018, 22 (28):  4475-4480.  doi: 10.3969/j.issn.2095-4344.0390 Abstract ( 333 )   PDF (1571KB) ( 129 )   Save BACKGROUND: Baicalin, as an important traditional Chinese medicine extract, has been extensively studied. Baicalin has been shown to promote the proliferation of human periodontal ligament fibroblasts (HPLFs) and differentiation into osteocytes, but the underlying molecular mechanism remains unclear. OBJECTIVE: To study the effect of baicalin on the proliferation of HPLFs and the underlying molecular mechanism. METHODS: Morphology of HPLFs was observed by primary cell culture and light microscopy. The growth-promoting effect of baicalin on HPLFs was detected by cell counting kit-8 assay. The effect of baicalin on the expression of proliferating cell nuclear antigen was detected by qRT-PCR. The inhibitory effect of baicalin on cell apoptosis was detected by qRT-PCR and flow cytometry. The level of bone morphogenetic protein 2 (BMP2) under baicalin treatment was tested by western blot assay and qRT-PCR. BMP2 antibody was employed to detect the influence of baicalin on the proliferation and apoptosis of HPLFs and the expression of BMP2 in HPLFs. RESULTS AND CONCLUSION: Compared with the control group, baicalin could significantly promote the proliferation of HPLFs, and up-regulate the mRNA expression level of proliferating cell nuclear antigen. Compared with the control group, baicalin could significantly up-regulate the mRNA level of Bcl-2 and down-regulate the mRNA level of Bad. Flow cytometry also showed that baicalin could inhibit cell apoptosis. Baicalin could up-regulate the protein and mRNA levels of BMP2 in a time- and concentration-dependent manner. The regulatory effect of baicalin was decreased after addition of BMP2 antibody. In summary, baicalin promotes proliferation of HPLFs probably via up-regulating BMP2 level. 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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Lung injury and inflammatory responses in a rabbit model of brain death Li Ling1, Xu Qian1, Wei Wan-hui1, Zhao Hui-jia1, Shi Yu-ying1, Liu Lu1, Wang Yan-feng1, Ye Qi-fa1, 2 2018, 22 (28):  4481-4486.  doi: 10.3969/j.issn.2095-4344.0367 Abstract ( 399 )   PDF (1399KB) ( 111 )   Save BACKGROUND: The quality of organs from donation after brain death is essential for the success of transplantation. Studies on the mechanism and protection of organ injury induced by brain death will provide references for the reasonable application of donor organs. OBJECTIVE: To investigate the changes in the liver morphology and inflammatory factors at different time points after brain death in rabbits, and to explore the mechanism of inflammatory factors in lung damage induced by brain death. METHODS: Forty rabbits were randomized into two groups (n=20 per group): sham operation group (tracheal intubation, femoral artery intubation, and sphenotresia), brain death group (low intracranial pressure to establish a state of brain death). The mean arterial pressure and heart rate in the two groups were recorded at postoperative 2, 6, 8 hours; the pathological changes of lung were observed by hematoxylin-eosin staining. The expression levels of inflammatory factors in the lung tissues including heat shock protein 27, intercellular adhesion molecule-1 and nuclear factor-κB were tested by RT-PCR and immunohistochemistry. The serum levels of interleukin-1, -6, and -8 were detected by ELISA. RESULTS AND CONCLUSION: There were no significant differences in the mean arterial pressure and heart rate at 2, 6 and 8 hours after brain death (P > 0.05). The serum levels of interleukin-1, -6, and -8 increased with time, and the differences were significant (P < 0.05). The mRNA level of heat shock protein 27 in the brain death group was significantly higher than that in the sham operation group (P < 0.05). Under light microscope, the alveolar cavity was intact at 2 hours after brain death, capillary hemorrhage, widened alveolar septum and inflammatory cell infiltration occurred at 6 hours after brain death, and inflammatory cells almost infiltrated the entire lung at 8 hours, which were consistent with the expression levels of inflammatory factors like intercellular adhesion molecule-1 and nuclear factor-κB in the brain death group. In summary, brain death can lead to lung damage, which gets worse with time, and the damage is related to the release of inflammatory factors. Moreover, the lung damage peaks at 8 hours after brain death. 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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Differences in the construction of thioredoxin-interacting protein overexpression model in INS-1 cells by two adenovirus infection methods Cao Zhu-jie, Feng Yan-jin, Li Dan, Wang Jin, Jiao Xiang-ying 2018, 22 (28):  4487-4492.  doi: 10.3969/j.issn.2095-4344.0839 Abstract ( 287 )   PDF (4821KB) ( 88 )   Save BACKGROUND: There are two methods for constructing the protein overexpression model in cells by adenovirus infection. In Method I, 3 mL of 1640 medium was supplemented 4 hours after cell infection by adenovirus, and 24 hours later, culture medium was refreshed with complete medium for further 48 hour culture, In Method II, 4 hours after cell infection by adenovirus, PBS was used to wash 1640 culture medium containing adenovirus, and then 4 mL of complete culture was added for further culture till 48 hours. The main difference between the two methods is that the adenovirus in Method I has a longer action time and a better infection effect, but the damage of the adenovirus itself to cells is more severe, and it disturbs the stability of the experimental model. The effect of adenovirus in Method II is short lasting, and the toxicity of adenovirus to cells is lower, but the efficiency of adenovirus infection is weaker, and the expression of target protein is weaker. OBJECTIVE: To find a more stable and efficient way to establish the thioredoxin-interacting protein (TXNIP) overexpression model by adenovirus infection in INS-1 islet β-cells. METHODS: The adenovirus vector (Ad-GFP) with green fluorescent protein (GFP) and TXNIP overexpressing adenovirus vector (Ad-TXNIP-GFP) with green fluorescent protein (GFP) were constructed. Three groups (normal, Ad-GFP, Ad-TXNIP-GFP) were transfected with methods I and II, respectively. RESULTS AND CONCLUSION: The GFP fluorescence intensity and the GFP positive percent in Method II were greater than those in the Method I. The cell morphology and survival rate in Method II were better than those in Method I. The expression of TXNIP at mRNA and protein levels in Method II was significantly higher than that in Method I. These findings show that the method II is more conducive to INS-1 cells in the construction of TXNIP overexpression model.  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 Duhuo Jisheng Decoction on PERK/Bip signaling pathway in cartilage of a rat model of knee osteoarthritis Chen Jun1, Wu Guang-wen2, Xu Hui-feng2, Zheng Chun-song3, Li Xi-hai3, Qiu Jian-qing3, Liu Shu-ru3, 2018, 22 (28):  4493-4500.  doi: 10.3969/j.issn.2095-4344.0391 Abstract ( 300 )   PDF (7260KB) ( 108 )   Save BACKGROUND: Preliminary study has shown that Duhuo Jisheng Decoction (DHJSD) is an effective prescription for osteoarthritis. But the underling mechanism remains unclear. OBJECTIVE: To observe the effect of DHJSD on the key regulating factors PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 in PERK/Bip signaling pathway in cartilage of rats with knee osteoarthritis, so as to explore its treatment mechanism. METHODS: After 1 week of acclimation, 80 Sprague-Dawley rats were randomly divided into normal group (n=20) and model rats (n=60). Osteoarthritis was induced by injecting 0.2 mL of 4% papain solution in both knees. The rat models were randomly divided into three groups, including model, treatment and positive control groups (n=20 per group). The normal and model groups received treatment of normal saline. The treatment and positive control groups were given DHJSD (9.3 g/(kg•d)) and glucosamine hydrochloride capsule 0.15 g/(kg•d) via gavage, respectively, for four courses (two weeks a course). After every two courses, the animals were sacrificed and the right tibial plateau was obtained. The mRNA and protein levels of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 were measured by RT-PCR and immunohistochemistry, respectively. RESULTS AND CONCLUSION: RT-PCR results showed that DHJSD and glucosamine hydrochloride capsule could time-dependently inhibit the mRNA expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 (P < 0.05 or P < 0.01). There was no significant difference between treatment and positive control groups (P > 0.05). Immunohistochemistry results found that DHJSD and glucosamine hydrochloride capsule could time-dependently inhibit the protein expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 (P < 0.05 or P < 0.01). There was no significant difference between treatment and positive control groups (P > 0.05). These results suggest that DHJSD can inhibit the apoptosis of chondrocytes caused by endoplasmic reticulum stress through regulating the PERK/Bip signaling pathway. 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 rabbit model of early femoroacetabular impingement and its morphological characteristics Xiong Ao, Luan Xin-yu, Lin Jian-jing, Kang Bin, Zeng Hui 2018, 22 (28):  4501-4506.  doi: 10.3969/j.issn.2095-4344.0852 Abstract ( 496 )   PDF (4864KB) ( 139 )   Save BACKGROUND: Femoroacetabular impingement (FAI) is a disease causing pain and dysfunction of hip joint in young active populations and has been paid more and more attention in recent years. However, its pathogenesis  and optimized treatment have not been fully understood. OBJECTIVE: To establish an animal model of early FAI in New Zealand rabbits and investigate the feasibility of this model for the study on FAI by morphological observation. METHODS: Twelve New Zealand rabbits were given unilateral femoral osteotomy with adduction 30° and external rotation 45° to establish the animal model of early FAI, and were then euthanized at 4 and 16 weeks in batches. Hip X-ray films were taken for comparison, the hip specimens were obtained for gross observation, then hematoxylin-eosin staining and toluidine blue staining were performed to observe the cartilage morphology in the area of impingement, and the cartilage degeneration was evaluated by Mankin scores. RESULTS AND CONCLUSION: Some low-density round areas were visible at the subchondral bone of the femoral head-neck conjunction in partial modeling femurs by X-ray. Gross specimen indicated the signs of cartilage injury at the impinging area. Histological observation also proved the cartilage degeneration at the femoral head-neck conjunction and acetabular labrum. The Mankin scores of the surgical side were significantly higher than those of the control side. Therefore, the animal model of early FAI in New Zealand rabbit is established successfully, which facilitates researches on the pathogenesis and treatment of FAI. 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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STEP61 negatively regulates amyloid beta-mediated ERK signaling pathway in Alzheimer’s disease cell model  Zhang Lin1, Yang Jing2, Liu Zan-hua1 2018, 22 (28):  4507-4512.  doi: 10.3969/j.issn.2095-4344.0796 Abstract ( 543 )   PDF (1261KB) ( 97 )   Save BACKGROUND: Striatal-enriched phosphatase 61 (STEP61) has been shown to play an important role in  synaptic plasticity. STEP is located at the postsynaptic terminal, and affects the formation of synaptic enhancement probably through the dephosphorylation and inactivation of key signal proteins such as MAPK kinase ERK1/2. OBJECTIVE: To explore the mechanism of STEP61 negatively regulating amyloid β-protein (Aβ)-mediated ERK signaling pathway in Alzheimer’s disease cell model. METHODS: There were three groups. Normal group was primary cortical neuron cells of C57BL/6 mice. In Alzheimer’s disease cell model group, 1 μmol/L Aβ1-42 (condensed state) was added into the cortical neuron cells for 1 hour culture to prepare Alzheimer’s disease cell model. In Aβ1-42 + RNAi group, STEP61 gene silencing was performed in Alzheimer’s disease cell model by sRNAi technique. The effect of STEP61 RNAi on ERK signaling pathway was detected by western blot assay. RESULTS AND CONCLUSION: The expression of STEP61 protein in the Alzheimer’s disease cell model group was increased by 223% compared with the normal group, and the ratio of phosphorylated STEP61 was reduced to (75.6 ± 4.6)% (P < 0.05). There was no significant difference in the ratio of STEP61 phosphorylation between Aβ1-42 + RNAi and Alzheimer’s disease cell model groups. Western blot analysis showed an evident decrease of pERK1/2 levels in the Alzheimer’s disease cell model group compared with the control group (P < 0.05). The expression of pERK1/2 protein in the Aβ1-42 + RNAi group was significantly higher than that in the Alzheimer’s disease cell model group (P < 0.05). These results suggest that STEP61 negatively regulates Aβ mediated ERK signaling pathway in the Alzheimer’s disease cell model, possibly by dephosphorylation. 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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Expression levels of Foxp3 and indoleamine 2,3-dioxygenase in mouse corneal allograft rejection Gong Yu-bo1, Liu Yong2, Zhao Hong-wei1, Xu Qian-qian1, Guo Hui-ling1 2018, 22 (28):  4513-4517.  doi: 10.3969/j.issn.2095-4344.0857 Abstract ( 482 )   PDF (1027KB) ( 110 )   Save BACKGROUND: Immunological rejection reaction is the main cause of corneal allograft failure and its pathogenesis is still unclear. Foxp3 and indoleamine 2,3-dioxygenase (IDO) are reported to be associated with transplantation immunity. OBJECTIVE: To study the roles of Foxp3 and IDO in mouse corneal allograft rejection. METHODS: Routine penetrating keratoplasty was performed with C57BL/6 mice as donors and 30 BALB/c mice as recipients. The recipient mice were randomly divided into normal group (n=6), allograft group (intraperitoneal injection of normal saline solution for 6 times at 0, 2, 4, 6, 8 and 10 days after allograft, n=12) and dexamethasone group (intraperitoneal injection of 10 mg/kg dexamethasone, n=12). The graft rejection was evaluated under slit lamp microscope. Expression level of interferon-γ in grafts was detected by immunohistochemical staining. The expression levels of Foxp3 and IDO mRNA in grafts were detected by real-time PCR. RESULTS AND CONCLUSION: The mean graft survival time in the allograft group was (14.833±1.472) days, and that in the dexamethasone group was (20.667±1.033) days (P < 0.05). The expression levels of interferon-γ as well as Foxp3 and IDO mRNA in grafts were significantly increased in the allograft group compared with the dexamethasone group (P < 0.05) at the 14th day postoperatively. These results suggest that Foxp3, IDO and interferon-γ play important roles in corneal allograft rejection; therefore, decreasing the expression levels of Foxp3, IDO and interferon-γ may contribute to inducing tolerance. 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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Expression levels of SIRT1 and oxidative stress injury markers in a mouse model of deep venous thrombosis Zhao Chong-yu, Wang Bing, Lou Zhen-kai, Li Xing-guo, He Wei, Zhao Xue-ling 2018, 22 (28):  4518-4524.  doi: 10.3969/j.issn.2095-4344.0301 Abstract ( 371 )   PDF (5867KB) ( 135 )   Save BACKGROUND: Silent information regulator1 (SIRT1) has been shown to inhibit oxidative stress and reduce arterial thrombosis. But changes in SIRT1 expression and oxidative stress in venous thrombosis remain unclear. OBJECTIVE: To further analyze the correlation of SIRT1 and oxidative stress in deep venous thrombosis. METHODS: Ninety C57 mice were randomized into three groups based on body mass: control, sham operation and model groups (n=30 per group). The mouse model of deep venous thrombosis was created by inferior vena cava stenosis, and the inferior vena cava tissues were gained at 24 hours after modeling. The thrombosis was observed by hematoxylin-eosin staining; the content of reactive oxygen species was assayed by membrane permeable fluorescent probe DCFH-DA and flow cytometry; contents of superoxide dismutase and malondialdehyde were tested; SIRT1 protein expression was detected by western blot assay. RESULTS AND CONCLUSION: Compared with the control and sham operation groups, in the modeling group, the contents of reactive oxygen species and malondialdehyde were significantly increased, and the level of superoxide dismutase was significantly decreased (P < 0.05). The expression level of SIRT1 in the modeling group was significantly lower than that in the control and modeling groups (P < 0.05). In summary, oxidative stress plays a significant part in the germination and growth of deep venous thrombosis, and the activity of SITR1 is inhibited during deep venous thrombosis formation. 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 electroacupuncture on the expression of N-cadherin, E-cadherin and P-cadherin in the facial neurons after facial nerve injury Gong Li-qiong, Cai Hong, Fei Jing, Li Lei-ji 2018, 22 (28):  4525-4531.  doi: 10.3969/j.issn.2095-4344.0845 Abstract ( 282 )   PDF (6688KB) ( 129 )   Save BACKGROUND: Preliminary studies have found that peripheral facial nerve injury can cause changes in the expression of adhesion molecules in neurons, but the mechanism of electroacupuncture in the treatment of facial nerve injury is unclear. OBJECTIVE: To observe the effect of electroacupuncture on the morphological changes of the facial neurons and the expression of N-cadherin, E-cadherin and P-cadherin in the neurons after peripheral facial nerve injury. METHODS: New Zealand white rabbits were randomized into control group (n=4), model group (n=24) and electroacupuncture group (n=24). Rabbits in the latter two groups were used to establish the crush injury model of superior buccal branch of the right facial nerve, and the electroacupuncture group rabbits were given the electroacupuncture at Jiache (ST6), Dicang (ST4), Yifeng (SJ17), Quanliao (SI18), Sibai  (ST2) and Yangbai (GB14) with dilatational wave (18-20 Hz, 1.5 V), and needle-embedding at Hegu (LI4) without electroacupuncture, 30 minutes, once daily. RESULTS AND CONCLUSION: In situ hybridization results suggested that E-cadherin mRNA and P-cadherin mRNA were negative in all groups, and N-cadherin mRNA was not expressed in the control group. The expression of N-cadherin mRNA was positive in the electroacupuncture and model groups after facial nerve injury. The number of N-cadherin mRNA positive cells in the electroacupuncture group was significantly higher than that in the model group (P < 0.05). Immunohistochemical staining results found that the expressions of N-cadherin, E-cadherin and P-cadherin were positive in the control group; the expression levels of the three cadherins decreased at 1 day after injury (except N-cadherin in the electroacupuncture group); the expression levels of N-cadherin and P-cadherin in the electroacupuncture group were higher than those in the model group at 1, 4 and 14 days after injury (P < 0.05); the expression levels of E-cadherin in the electroacupuncture group was significantly higher than that in the model group (except the first day) (P < 0.05). Hematoxylin-eosin staining showed that in the model group, there was facial nerve neuron degeneration followed by similar neuronal death, and the neuron morphology began to recover at 21 days after injury. Neuron morphological changes in the electroacupuncture group were less than those in the model group, and neurons began to recover at 14 days after injury. The neurons in the two groups were restored at 28 days after injury. Our findings reveal that facial nerve injury can induce the expression of adhesion molecules, and electroacupuncture can reduce the degeneration of facial neurons and promote the recovery of neurons, which may be achieved by increasing the expression of adhesion molecules in the facial neurons. 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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Imaging characteristics of near-infrared fluorescent dye IR-780 in clear cell renal cell carcinoma Zhang Wei1, Qu Zhi-yi1, Liu Xiao-yang1, Wang Fu-li2, Ye Yong-li2, Yan Fei2, Wei Di2, Yuan Jian-lin2 2018, 22 (28):  4532-4536.  doi: 10.3969/j.issn.2095-4344.0332 Abstract ( 445 )   PDF (3963KB) ( 122 )   Save BACKGROUND: The near-infrared fluorescent dyes that directly bind to tumor cells have been developed, and IR-780 stands out. OBJECTIVE: To evaluate the specific imaging effect of near-infrared fluorescent dye IR-780 in clear cell renal cell carcinoma (ccRCC). METHODS: Human ccRCC cells 786-O, ACHN and normal human embryonic kidney 293T cells in logarithmic growth phase were incubated with 20 μmol/L IR-780 for 30 minutes. The localization of the dye in ccRCC was observed by laser confocal microscopy. Human ccRCC cells 786-O in logarithmic phase was incubated with 20 μmol/L IR-780 for 30 minutes. The localization of the dye in mitochondria and lysosomes was observed under confocal laser scanning microscope. 10, 100, 1 000 and 10 000 DAPI-stained human ccRCC cells 786-O were respectively added to human blood samples. The mononuclear cells in each group were isolated and cultured with 20 μmol/L IR-780 for 30 minutes. DAPI and IR-780 double positive cells were observed under confocal laser scanning microscope. RESULTS AND CONCLUSION: IR-780 possessed the ability to visualize a variety of ccRCC cells, but not for normal renal epithelial cells.  IR-780 had obvious overlapping staining in lysosomes or mitochondria, indicating its selective accumulation in mitochondria and lysosomes in the organelles. IR-780 could detect a small amount of ccRCC in the blood. These results suggest that the near-infrared fluorescent dye IR-780 can specifically accumulate in ccRCC cells, which can be used for specific diagnosis of ccRCC. 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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Inhibition of high mobility group protein attenuates spinal astrocyte injury in rats after oxygen glucose deprivation/reoxygenation  Song Jun-lai1, Li Man2, Sun Lin1, Ma Xun1, Lü Cong1, He Ya-jun1 2018, 22 (28):  4537-4543.  doi: 10.3969/j.issn.2095-4344.0836 Abstract ( 317 )   PDF (1612KB) ( 134 )   Save BACKGROUND: Many investigations focus on the inhibition of high mobility group protein (HMGB1) for improving the functional recovery of spinal cord in mice, but how to culture spinal cord astrocytes in vitro and inhibit the expression of HMGB1 for attenuating oxygen glucose deprivation/reoxygenation (OGD/R) injury is rarely reported. OBJECTIVE: To investigate the effect of HMGB1 inhibition on spinal cord astrocytes after OGD/R in vitro. METHODS: Astrocytes were isolated from rat spinal cord, and were then cultured and identified. (1) The rat primary spinal cord astrocytes were subjected to 6-hour OGD, followed by 6-, 12-, and 24-hour reoxygenation, respectively. The rat primary spinal cord astrocytes cultured in the normal medium were used as controls. (2) The rat spinal cord astrocytes were divided into five groups: control group (normal medium); 6-hour OGD/24-hour R group; 6-hour OGD/24-hour R plus HMGB1 shRNA group; 6-hour OGD/24-hour R plus non-targeting shRNA group; 6-hour OGD/24-hour R plus ethyl pyruvate group. The expression and release of HMGB1 in astrocytes were determined by western blot assay and ELISA. The cell injury and survival rate were assessed by lactate dehydrogenase and MTT assay. The morphological changes of the cells were observed under light microscope. RESULTS AND CONCLUSION: The expression and release of HMGB1 protein was increased after OGD/R (P < 0.01), and reached the highest value at reoxygenation for 24 hours (P < 0.01). The expression level of HMGB1 in the HMGB1 shRNA and ethyl pyruvate groups was signficnatly decreased, suggesting that specific RNA interference and ethyl pyruvate could effectively inhibit the expression of HMGB1. The leakage rate of lactate dehydrogenase was increased and cell survival rate was decreased after OGD/R, and cell injury was the most serious at reoxygenation for 24 hours (P < 0.01). Compared with the 6-hour OGD/24-hour R group, the leakage rate of lactate dehydrogenase in the HMGB1 shRNA and ethyl pyruvate groups was significantly reduced. There were cell swelling, decomposition and other damage changes after OGD/R, while HMGB1 shRNA and ethyl pyruvate RNA could significantly reduce cell injury. Our findings imply that HMGB1 plays an important role in the occurrence and development of spinal cord astrocyte injury, and inhibition of HMGB1 can alleviate spinal cord astrocyte injury in rats after OGD/R. 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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Nicotinamide N-methyl transferase gene polymorphism and its correlation with the athletic ability of handball players Chen Wei1, Li Jiang-hua2, Gu Jian-ren1, Chen Feng1, Xia Hua1, Zhang Bin1, Jiao Qi-bin1, Zheng Feng3 2018, 22 (28):  4544-4549.  doi: 10.3969/j.issn.2095-4344.0321 Abstract ( 274 )   PDF (1162KB) ( 99 )   Save BACKGROUND: Studies confirm that niacinamide N-methyl transferase gene (NNMT) polymorphism is associated with human body aerobic exercise capacity. Handball is a kind of mixed oxygen sports, accounting for a proportion of anaerobic exercise in competition or peacetime training, mainly in aerobic exercise. OBJECTIVE: To explore a NNMT gene single nucleotide polymorphism locus rs2256292 associated with the athletic ability of handball players. METHODS: The NNMT genetic polymorphism locus rs2256292 in 28 athletes from Chinese national male handball team (experimental group) and 53 male students (control group) was tested by polymerase chain reaction-ligase detection reaction. The relative VO2max and body constituents index of skeletal muscle, body fat percentage were also tested. RESULTS AND CONCLUSION: Frequencies of G allele and GG genotype in the experimental group were significantly higher than those in the control group (P < 0.05). Among the 28 athletes, the relative VO2max in the GG genotype group was higher than that in the CG genotype group (P < 0.05). Among the 53 general students, there were no differences in relative VO2max among genotypes. Among the 28 athletes, the skeletal muscle percentage in the GG genotype group was significantly higher than that in the CC and CG genotype groups (P < 0.05). Among the 53 general students, the skeletal muscle percentage in the GG genotype group was significantly higher than that in the CG genotype group (P < 0.05). In addition, there were no differences in the body fat percentage among genotypes in athletes or general students. These results suggest that the NNMT genetic polymorphism locus rs2256292 is significantly correlated with the athletic ability of handball players, with the athletes being more likely to have G allele and carriers of GG genotype having stronger athletic ability. 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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Different methods for establishing rabbit models of knee osteoarthritis: rational option and characters Peng Xu, Xiong Hua-zhang, Yang Ji-bin, You Qi, Jin Ying, Zhang Jun, Ge Zhen, Liu Yi 2018, 22 (28):  4550-4556.  doi: 10.3969/j.issn.2095-4344.0330 Abstract ( 740 )   PDF (1127KB) ( 137 )   Save BACKGROUND: Animal models have been widely applied in the studies on osteoarthritis. Rabbits are the most commonly used animal models. In many modeling methods, inductive model is preferred because of good stability, high efficiency, and repeatability. OBJECTIVE: To analyze the research status and application of rabbit models of knee osteoarthritis at home and abroad in recent years. METHODS: A computer-based search of PubMed, Web of Science, CNKI, WanFang, and VIP databases was performed. The keywords were “animal model, osteoarthritis knee, rabbit” in English and Chinese, respectively. The articles concerning rabbit models of knee osteoarthritis published between August 1983 and December 2017 were retrieved, including reviews, basic researches, and clinical studies. The articles were screened preliminarily by reading the topic and abstract of the screening, and irrelative articles were excluded. Finally 52 articles were enrolled for review according to the inclusion and exclusion criteria. RESULTS AND CONCLUSION: Animal models of osteoarthritis are mainly divided into spontaneous, inducible, gene knockout types. Inductive models are the most commonly used models in clinical trials, including surgical and non-surgical methods. The anterior cruciate ligament transaction is the commonly used surgical methed, and immobilization model is the most common non-surgical method. The induced model can quickly and steadily cause the pathophysiological changes similar to the degeneration of human knee joint, and the models are mainly used for the studies on the drugs, pathogenesis, and signaling pathways of osteoarthritis. A variety of modeling methods can successfully establish osteoarthritis model, but each has their own advantages and disadvantages, so we should make a reasonable choice in accordance with the experimental design and experimental conditions. 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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Animal models of stress urinary incontinence: selection of appropriate construction methods of research objective-related animal models Wang Tian-xing1, Chen Yue-lai2, Yin Ping3, Xu Shi-fen3, Zheng Hui-min3 2018, 22 (28):  4557-4561.  doi: 10.3969/j.issn.2095-4344.0252 Abstract ( 297 )   PDF (1003KB) ( 149 )   Save BACKGROUND: Animal models of stress urinary incontinence (SUI) can simulate the anatomical structure and pathological state of pelvic floor in female patients with SUI. Animal models are important for studies on the pathogenesis and therapeutic effect of SUI. OBJECTIVE: To summarize the SUI modeling methods and research ideas, reveal the status and progress of establishing an animal model of SUI, and compare the advantages and disadvantages of various modeling methods, thereby providing the basis for ideal animal model construction and providing favorable reference for clinical research. METHODS: Databases of CNKI and PubMed were retrieved using computer with the search time of January 2000 to August 2017. The keywords were “stress urinary incontinence and animal model” in English and Chinese, respectively. The articles addressing animal models rather than mechanism were excluded. Pathogenesis, establishment and pathological characteristics of animal models of SUI were summarized. RESULTS AND CONCLUSION: A total of 128 articles were retrieved, the repetitive and obsolete articles were excluded, and then 37 eligible articles were included. The animal models of SUI mainly relay on the mechanical damage of the pelvic floor support organization. All kinds of models can only focus on some pathological causes of a disease, or pathological changes at a certain stage. Moreover, the maintenance time, advantages and disadvantages are different. So researchers must choose appropriate animal models according to their own requirements. 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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Biomarkers for lumbar disc degeneration: how to solve the influences of specificity, sensitivity and covariates  Guo Mei-yu, Li Zhong-hai 2018, 22 (28):  4562-4567.  doi: 10.3969/j.issn.2095-4344.0369 Abstract ( 496 )   PDF (1026KB) ( 114 )   Save BACKGROUND: Serum biomarkers have been shown to be helpful for the diagnosis and treatment of lumbar disc degenerative diseases. Understanding the pathogenesis and development process of lumbar disc degenerative diseases through biomarkers is of great significance for the clinical diagnosis, prevention and treatment of lumbar disc degenerative diseases. OBJECTIVE: To review the literature concerning the biomarkers for lumbar disc degenerative diseases to further understand the pathogenesis and help to select new treatment methods. METHODS: SinoMed, VIP, WanFang and PubMed databases were retrieved for the literature published from January 1990 to May 2018 using the keywords of “intervertebral disc degeneration; biomarkers” in English and Chinese, respectively. Relevant articles were screened according to the inclusion and exclusion criteria. The biomarkers and clinical application of lumbar disc degenerative diseases were summarized. RESULTS AND CONCLUSION: A total of 206 articles were collected and 32 eligible articles were enrolled after excluding repetitive studies. Various cytokines such as interleukin, interferon, tumor necrosis factor, vascular endothelial growth factor and chemokine were found to be related to lumbar disc degenerative diseases. The biomarkers related to degenerative disc diseases can be used for early diagnosis, disease progression and evaluation of therapeutic effect. However, the development of biomarkers is still limited, such as poor sensitivity and specificity of biomarkers for early lumbar disc degenerative changes, and influence of disease covariates (age, body mass index and depression), so a further investigation is needed. 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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Biological role of myonectin and its correlation with exercise Li Lin, Yu Liang 2018, 22 (28):  4566-4573.  doi: 10.3969/j.issn.2095-4344.0266 Abstract ( 451 )   PDF (1073KB) ( 113 )   Save BACKGROUND: Myonectin (C1q tumor necrosis factor-α-related protein isoform 15) is a novel bioactivator secreted by skeletal muscle and is the 15th member of CTRP family. Myonectin is specially expressed in skeletal muscle and its expression is affected by the type of skeletal muscle fiber, nutritional conditions, other hormones, and cytokines. OBJECTIVE: To explore the relationship between myonectin and exercise so as to provide the basis for the treatment of chronic diseases. METHODS: A computer-based search of CNKI and PubMed databases was performed for relevant articles published from 1999 to 2017 using the keywords of “myokine, myonectin, erythroferrone” in Chinese and English, respectively. Finally 32 eligible articles were included in result analysis in accordance with the inclusion and exclusion criteria. RESULTS AND CONCLUSION: Myonectin is released into bloodstream through endocrine or paracrine and works on adipose tissue and liver, involved in adipose tissue and liver lipid metabolism regulation. Nevertheless, myonectin can increase the expression of fatty acid uptake protein gene, promote the uptake of fatty acids and reduce the level of free fatty acids in serum. Notably, myonectin participates in obesity and insulin resistance and correlates with skeletal muscle energy metabolism. It is also involved in erythropoiesis during the regulation of iron metabolism, and therefore, myonectin is closely related to metabolic diseases. 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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Correlation between gene polymorphism of macrophage migration inhibitory factor and disease susceptibility: recognition, target and significance Li Jing1, 2, Peng Yun3, 4, Bao Fu-kai2, 3, 4, 5, 6, Liu Ai-hua3, 4, 5, 6, 7 2018, 22 (28):  4574-4579.  doi: 10.3969/j.issn.2095-4344.0299 Abstract ( 270 )   PDF (1097KB) ( 121 )   Save BACKGROUND: Changes in the expression levels of macrophage migration inhibitory factor (MIF) occur in a variety of diseases, and the gene polymorphism of MIF is related to the disease susceptibility and severity. The basic physicochemical properties of MIF, and correlation of MIF gene polymorphism with disease susceptibility are summarized. OBJECTIVE: To analyze the mechanisms of MIF in various diseases. METHODS: Databases of PubMed, Sino Med and CNKI were searched by computer for the articles concerning MIF and diseases using the keywords of “MIF, disease, polymorphism” in English and Chinese, respectively. Finally 58 eligible articles were included. RESULTS AND CONCLUSION: MIF is involved in various diseases such as autoimmune disease, inflammatory disease and cancer. However, MIF plays different roles in various systemic diseases, for example, MIF promotes the secretion of tumor necrosis factor α in adiposities inducing insulin resistance. It also interferes with insulin-mediated NO release from endothelial cells and participates in the development of insulin resistance in endothelial cells. MIF counteracts glucocorticoid effects, restores macrophage factor expression and activates T lymphocytes, which plays an important role in the regulation of inflammatory response. Therefore, to understand the interaction between MIF gene polymorphism and systemic diseases is conducive to the development of basic research on MIF in various systemic diseases. It is of great significance to find MIF as a therapeutic target for various diseases. 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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Roles of nerve growth factor and brain-derived neurotrophic factor and their precursors in nervous system Dai Yun-fei1, Wang Tong-tong1, Ma Wei1, Yang Jin-wei1, 2, Wang Xian-bin1, Zhang Tong2, Su Ping3, Guo Jian-hui2, Li Li-yan1 2018, 22 (28):  4580-4586.  doi: 10.3969/j.issn.2095-4344.0300 Abstract ( 405 )   PDF (780KB) ( 79 )   Save BACKGROUND: More than 20 neurotrophic factors have been found. Among them, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are the most widely distributed growth factors in the central nervous system, and they are responsible for the growth, development and neuronal plasticity of the central nervous system and even of the entire brain. OBJECTIVE: To fully understand NGF and BDNF, learn more about their role in the repair of nervous system damage, and to review the research progress in NGF and BDNF and their precursors. METHODS: A computer-based online retrieval of PubMed and CNKI databases was performed to search relevant papers about neurotrophic factors, NGF and BDNF. The keywords were “nerve growth factor (NGF), proform of nerve growth factor (proNGF), brain-derived neurotrophic factor (BDNF), precursor for brain-derived neurotrophic factor (proBDNF), neurotrophic factor, neurotrophin receptor” in English and Chinese, respectively. RESULTS AND CONCLUSION: NGF has the biological function of protecting neurons after peripheral nerve injury. The precursor of NGF simultaneously forms trimers of signal transducers with receptors p75NTR and Sortilin and induces apoptosis and death of nerves post-nerve injury. PDNF preferentially binds to TrkB to produce biological effects that promote synaptic growth or support the survival and differentiation of neurons. The precursor of BDNF preferentially binds p75NTR and Sortilin to trigger neuronal apoptosis. With the deepening of the research, it has been found that the precursors of NGF and BDNF exhibit the opposite biological effects on the mature NGF and BDNF, but the specific mechanism of action remains unclear. 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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Expression and role of wnt signaling pathway in neurogenesis and neurodegeneration Wang Xian-bin1, Tang Hong-yan2, Li Xing-tong1, Ma Wei1, Yang Jin-wei1, 3, Dai Yun-fei1, Zhang Tong3, 2018, 22 (28):  4587-4592.  doi: 10.3969/j.issn.2095-4344.0241 Abstract ( 241 )   PDF (1085KB) ( 96 )   Save BACKGROUND: Wnt signaling pathway plays an important role in the nervous system as a multi-link and multi-site open pathway. Therefore, it is important to understand its relationship with the pathophysiology and treatment of neurological diseases. OBJECTIVE: To analyze the mechanism of Wnt signaling pathway in nervous system diseases. METHODS: PubMed and SinoMed were searched for the relevant articles concerning Wnt signaling pathway and nervous system diseases. The key words were “Wnt signaling pathway, nervous system disease” in English and Chinese, respectively. Finally, 62 pertinent articles were included for result analysis. RESULTS AND CONCLUSION: Wnt signaling pathway is involved in Alzheimer’s disease, Parkinson’s disease and cerebrovascular disease. However, its role is different in different neurological diseases. For example, the dysfunction of Wnt signaling pathway leads to the disability of neuronal formation, which accelerates the progression of Alzheimer’s disease. Besides, Wnt signaling pathway regulates the proliferation of dopaminergic neurons, cell survival and synaptogenesis, thus affecting the development of Parkinson’s disease. Therefore, understanding the interaction between ligands, receptors and genes in Wnt signaling pathway in nervous system diseases is conducive to the development of Wnt signaling in the basic research of neurological diseases, and is of great significance for searching for the treatment targets of nervous system diseases. 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