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08 November 2020, Volume 24 Issue 31 Previous Issue    Next Issue

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Rev-erbα’s effect on osteoblastogenesis of mouse bone marrow mesenchymal stem cells  Zhang Shuang, Xu Xiaomei, Zeng Yang, Yuan Xiaoping, Lin Fuwei 2020, 24 (31):  4921-4926.  doi: 10.3969/j.issn.2095-4344.2117 Abstract ( 524 )   PDF (785KB) ( 116 )   Save BACKGROUND: The orphan nuclear receptor Rev-erbα has been demonstrated to play important roles during bone metabolism. However, the specific mechanism underlying osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is still unclear. OBJECTIVE: To explore whether the orphan nuclear receptor Rev-erbα participates in the osteogenesis process of BMSCs in mice. METHODS: Mouse BMSCs were isolated and cultured by the whole bone marrow adherence method. And the lentivirus vector carrying the Rev-erbα gene was constructed and then transfected into the BMSCs. Real-time PCR was conducted to detect the mRNA level of Rev-erbα at 48 hours after transfection. There were three groups in the experiment: an experimental group with BMSCs transfected with lentiviral vectors containing overexpressed Rev-erbα gene and EGFP gene, a positive control group with BMSCs transfected with empty lentiviral vectors containing only EGFP gene, and a negative control group. Then the BMSCs were induced to differentiate into osteoblasts after the transfection and we detected the mRNA level of alkaline phosphatase, osteopontin, and osteocalcin using real-time PCR at 0, 7, and 14 days after induction. RESULTS AND CONCLUSION: The lentivirus vector carrying Rev-erbα gene was successfully transfected into BMSCs, and expressed stably. An increase in the mRNA levels of alkaline phosphatase and osteopontin was detected in each group when cultured in osteogenic medium; however, there was no significant difference between groups. The mRNA level of osteocalcin was also increased in each group, and moreover, the mRNA level in the experimental group was significantly lower than that in the two control groups (P < 0.05). These results indicate that Rev-erbα transfection reduces the osteogenic ability of BMSCs, and the expression of osteocalcin, a late-stage osteogenic marker, is inhibited, indicating that the late-stage osteogenesis of BMSCs may be influenced by Rev-erbα. Figures and Tables | References | Related Articles | Metrics

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Potential molecular targets and therapeutic mechanisms underlying transplantation of autologous bone marrow stem cells for the treatment of spinal cord injury based on bioinformatics Xun Chong, Wang Qiang, Li Changzhou, Liu Xiaofeng 2020, 24 (31):  4927-4933.  doi: 10.3969/j.issn.2095-4344.2131 Abstract ( 507 )   PDF (772KB) ( 126 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have achieved good clinical effect in the treatment of spinal cord injury. However, most of the existing studies focus on the pathophysiological changes, and less is reported on the molecular regulatory mechanism involved. OBJECTIVE: To study the potential molecular targets and signal pathway related to the use of bone marrow mesenchymal stem cells in the treatment of spinal cord injury based on bioinformatics analysis. METHODS: The microarray data of bone marrow mesenchymal stem cells for spinal cord injury treatment were retrieved from GEO database and then mapped to the background network of spinal cord injury obtained from CTD database. Finally, the differentially expressed genes were obtained. A protein-protein interaction network constructed by sting was used to analyze its network characteristics using Cytoscape and obtain the core molecular targets in the process. Finally, the molecular functions and signal pathways of the above molecular targets were analyzed by functional enrichment of the above genes on the website of WebGestalt. RESULTS AND CONCLUSION: According to the data of GEO chip, there are 126 core molecular targets in the treatment of spinal cord injury by bone marrow mesenchymal stem cells, 60 of which have been reported in CTD database. According to the results of sting and Cytoscape, there are 57 nodes in the protein-protein interaction network, among which 10 core targets are MAPK3, tumor necrosis factor, interleukin 6, CASP3, vascular endothelial growth factor A, MAPK1, CCL2, interleukin 1B, proto-oncogene protein, insulin-like growth factor 1. In this process, the main regulatory pathways are interleukin 17, MAPK, tumor necrosis factor, Toll-like receptor and nod-like receptor. The two most important aspects of this process are the inhibition of inflammatory response mediated nerve damage and the nutritional repair of damaged nerve. Figures and Tables | References | Related Articles | Metrics

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Fabrication of prevascularized osteogenic differentiated cell sheet based on human bone marrow mesenchymal stem cells and human umbilical vein endothelial cells Chen Jia, Yang Yiqiang, Hu Chen, Chen Qi, Zhao Tian, Yong Min, Ma Dongyang, Ren Liling 2020, 24 (31):  4934-4940.  doi: 10.3969/j.issn.2095-4344.2129 Abstract ( 354 )   PDF (1065KB) ( 139 )   Save BACKGROUND: How to make the engineered bone tissues survive and function after transplantation, the precondition is to have sufficient and effective blood supply, which is one of the main obstacles in the clinical application of the engineered bone tissue. OBJECTIVE: To explore a new method to acquire vascular networks in vitro to realize vascularized engineered bone tissues. METHODS: Human bone marrow mesenchymal stem cells were cultured on a cell culture dish at a high density 9×104/cm2 for 14 days to form a thick cell sheet. The morphology of the cells was observed under a microscope and cells were stained with alkaline phosphatase and alizarin red. The human umbilical vein endothelial cells at 5×104/cm2 were seeded onto the surface of human bone marrow mesenchymal stem cells sheet for 7 days. The formation of vascular network and the characteristics of the sheet were evaluated by microscopic observation, CD31 immunofluorescence staining and type I collagen immunofluorescence staining.   RESULTS AND CONCLUSION: (1) Human bone marrow mesenchymal stem cells proliferated fast and formed an intact sheet, and the sheet cultured in osteogenic-induced medium showed positive staining of alkaline phosphatase and alizarin red. (2) Inverted phase contrast microscope showed that human umbilical vein endothelial cells migrated and aligned arrangement on the osteogenic differentiated cell sheet. The immunofluorescent images for CD31 revealed a progressive process of lumen formation. (3) Positive type I collagen was observed in the pre-vascularized osteogenic differentiated cell sheet. (4) These results indicate that the osteogenic differentiated cell sheet can be formed by continuously culturing. The prevascularizing method using coculture model of culturing endothelial cells onto an osteogenic differentiated cell sheet provides a theoretical guidance for optimizing the construction of prevascularized bone tissue. Figures and Tables | References | Related Articles | Metrics

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Calcined bone/chitosan composite promotes osteogenic differentiation of bone marrow mesenchymal stem cells in Sprague-Dawley rats   Liao Jian, Huang Xiaolin, Zhou Qian, Huo Hua, Qi Yuhan, Wu Chao, Shi Qianhui, Yang Tongjing, Liao Yunmao, Liang Xing 2020, 24 (31):  4941-4947.  doi: 10.3969/j.issn.2095-4344.2137 Abstract ( 430 )   PDF (890KB) ( 92 )   Save BACKGROUND: Our previous research has confirmed that the microstructure of calcined bone/chitosan composite is similar to that of natural bone tissue, safe and non-toxic, with good compressive strength, and can promote the proliferation and adhesion of osteoblasts. Therefore, it is necessary to investigate the effect of the composite on the osteogenic differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To investigate the effect of calcined bone/chitosan composite on osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: The calcined bone/chitosan composite was co-cultured with bone marrow mesenchymal stem cells of Sprague-Dawley rats for osteogenic induction in vitro. At 1, 2 and 3 weeks after osteogenic induction, the expression of genes related to osteogenic differentiation was detected by real-time fluorescence quantitative PCR. RESULTS AND CONCLUSION: The calcined bone/chitosan composites can promote the proliferation and adhesion of bone marrow mesenchymal stem cells and promote the expression of osteogenic genes alkaline phosphatase, type I collagen, and osteocalcin, which can induce the osteogenic differentiation of bone marrow mesenchymal stem cells.  Figures and Tables | References | Related Articles | Metrics

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MicroRNA expression profiles of chondrocytes in osteoarthritis induced by stromal cell derived factor 1  Wang Guoliang, Li Yanlin, Xiang Yaoyu, Jia Di, Li Canzhang, He Lu 2020, 24 (31):  4948-4953.  doi: 10.3969/j.issn.2095-4344.2107 Abstract ( 392 )   PDF (890KB) ( 78 )   Save BACKGROUND: Osteoarthritis is a complex disease caused by many factors, but its pathogenesis is yet unknown. Intensive molecular genetic research has indicated that noncoding RNA may be a potential transcriptional regulatory factor in osteoarthritis development. Exploration on differentially expressed miRNAs between osteoarthritis and normal tissue gives the clue to understand the molecular mechanism of osteoarthritis, providing a new cue for the diagnosis and treatment of osteoarthritis. OBJECTIVE: To discuss the changes of miRNA expression profile of chondrocytes stimulated by stromal cell derived factor 1 in osteoarthritis, and provide an experimental basis for delaying articular cartilage degeneration at the genetic level. METHODS: Residual cartilage samples from 10 patients with knee osteoarthritis who underwent total knee replacement were collected for culture of chondrocytes. The cells were then randomized into two groups: experimental group was cultured in high-glucose DMEM medium containing 10% fetal bovine serum and penicillin with the addition of 100 μg/L stromal cell derived factor 1, and control group was cultured in the medium with no other induction. At 48 hours after cell culture, the miRNA expression profiles of chondrocytes were detected, and the results were verified by fluorescence quantitative RT-PCR. The study protocol was performed in accordance with the Regulations on the Administration of Medical Institutions. All study patients gave written informed consent before collection of cartilage tissue. RESULTS AND CONCLUSION: After preliminary screening, 84 microRNAs were altered, of which 70 were up-regulated and 14 were down-regulated. Seven microRNAs with significant changes (miR-146a-5p, miR-124-3p, miR-130a-3p, miR-185-5p, miR-221-3p, miR-126-3p) were selected by gene chip for qRT-PPCR experiment, which indicated that the qRT-PCR results of miR-146a-5p, miR-124-3p and miR-126-3p were consistent with the results of gene chip. Therefore, induction with stromal cell derived factor 1 makes the expression profile of circulating miRNA change a lot. miR-146a-5p, miR-124-3p and miR-126-3p may be related to the stromal cell derived factor 1/CXCR4 signaling pathway by which stromal cell derived factor 1 can stimulate osteoarthritis. Figures and Tables | References | Related Articles | Metrics

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Relationship between mitochondrial autophagy and chondrogenesis of bone marrow mesenchymal stem cells Li Yuanqi, Lin Hai, Luo Hongrong, Zhang Xingdong 2020, 24 (31):  4954-4960.  doi: 10.3969/j.issn.2095-4344.2147 Abstract ( 563 )   PDF (849KB) ( 58 )   Save BACKGROUND: It has been found that mitochondrial autophagy plays an important role in cartilage defect repair. Therefore, it is necessary to study the influence and regulatory method of mitochondrial autophagy on the chondrogenesis of bone marrow mesenchymal stem cells, which will lay a foundation for further elucidating the cartilage-inducing mechanism and understanding how biomaterials regulate the differentiation and fate of bone marrow mesenchymal stem cells. OBJECTIVE: To establish and verify characterization methods of mitochondrial status and degree of autophagy, to observe the relationship of mitochondrial autophagy with chondrocyte development and the differentiation of bone marrow mesenchymal stem cells into cartilage. METHODS: Chondrocytes from neonatal neonates, 1-month-old rabbits, 18-month-old rabbits and neonatal neonatal rabbit bone marrow mesenchymal stem cells were separated and cultured. According to the instructions in the kit, Mito Tracker Red staining was conducted. The second-generation bone marrow mesenchymal stem cells were selected, and cultured with complete cartilage induction medium. Mitochondrial marker Mito Tracker Red staining was performed on days 1, 3, and 7 according to the kit instructions. The bone marrow mesenchymal stem cells of passage 2 were selected and cultured with complete cartilage induction medium for 24 hours. The induction group was added with 10 μmol/L autophagy inducer rapamycin for 10 hours, and the inhibition group was added with 5 μmol/L autophagy inhibitor chloroquine for 10 hours. After that, it was replaced with complete cartilage culture medium, and stained with mitochondrial autophagy kit Mitophagy Detection Kit on days 1, 3, and 7 to observe the situation of mitochondrial autophagy. RESULTS AND CONCLUSION: (1) Mitochondrial staining of chondrocytes from rabbits at different ages: The mitochondrial number in chondrocytes from young rabbits was higher than that from old rabbits. (2) Mitochondrial staining of bone marrow mesenchymal stem cells and newborn rabbit chondrocytes: The mitochondrial morphology in chondrocytes was spot dispersion, while that in bone marrow mesenchymal stem cells was linear distribution. (3) Mitochondrial staining during bone marrow mesenchymal stem cells differentiation: With the increase of induction culture time, the mitochondrial morphology in stem cells changed from the tubular network structure to a spotted situation, and the microfilament structure which promotes the formation of autophagy gradually decreases during the bone marrow mesenchymal stem cells differentiation. (4) The effect of mitochondrial autophagy on chondrogenic differentiation of bone marrow mesenchymal stem cells: Rapamycin promotes the mitochondrial autophagy pathway, and further promotes the chondrogenic differentiation of bone marrow mesenchymal stem cells.   Figures and Tables | References | Related Articles | Metrics

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Fetal bovine serum exosomes promote the proliferation of osteoblasts  Xu Huijun, Zhang Mi, Shi Dongmei, Wu Saixuan, Lu Ying, Dong Ming, Niu Weidong 2020, 24 (31):  4961-4965.  doi: 10.3969/j.issn.2095-4344.2122 Abstract ( 431 )   PDF (615KB) ( 39 )   Save BACKGROUND: Exosomes have been shown to promote bone regeneration, but whether extracting exosomes from fetal bovine serum can promote osteogenesis remains controversial. OBJECTIVE: To observe the effect of fetal bovine serum exosomes on the proliferation of osteoblasts, so as to provide new ideas for treating bone destruction. METHODS: Exosomes were extracted from fetal bovine serum by ultracentrifugation method. Electron transmission microscopy and western blot assay were used to verify whether the exosomes were successfully extracted. MC3T3-E1 cells were interfered with 10 mg/L fetal bovine serum. The effect of exosomes on the proliferation of osteoblasts was detected by cell counting kit-8 assay. The effects of exosomes on the expression of osteogenic markers bone morphogenetic protein-2 and osteopontin protein were detected by western blot assay. MC3T3-E1 cells cultured in the fetal bovine serum without exosomes were as control group. RESUITS AND CONCLUSION: The typical lipid bilayer membrane structure of exosomes was observed, and the size was between 30-150 nm. The exosome surface marker factor CD81 was positive, while the microcapsule surface marker CD40 was negative. The results of cell counting kit-8 assay showed that the proliferative capacity in the exosomes group was significantly higher than that in the control group (P < 0.05). The expression levels of bone markers bone morphogenetic protein-2 and osteopontin in the exosome group were significantly higher than those in the control group (P < 0.05). The exosomes extracted from fetal bovine serum can promote the proliferation of osteoblasts, providing a new idea for the clinical treatment of bone destruction. Figures and Tables | References | Related Articles | Metrics

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Co-transfection by Nell-1 and Noggin-shRNA promotes osteoblast differentiation of adipose derived mesenchymal stem cells Liu Hao, Liu Jun, Rui Yongjun, Tang Fenglin, Lu Miao, Ding Tao 2020, 24 (31):  4966-4970.  doi: 10.3969/j.issn.2095-4344.2116 Abstract ( 339 )   PDF (665KB) ( 112 )   Save BACKGROUND: Nell-1 can induce osteogenic differentiation and accelerate bone regeneration. However, little is reported about co-transfection by Nell-1 and Noggin-shRNA. OBJECTIVE: To observe the effects of simultaneously down-regulating Noggin and up-regulating Nel-like molecule-1(Nell-1) on osteogenic differentiation of adipose derived mesenchymal stem cells (ADSCs) in vitro.   METHODS: ADSCs were isolated and cultured in vitro from adipose tissue of healthy rats, and divided into three groups. ADSCs in control group were transfected with lentiviral (Lv)-enhanced green fluorescent protein. In Nell-1 group, the cells were transfected with Lv-Nell-1. And in co-transfection group, the cells were co-transfected with Lv-Nell-1 and Lv-Noggin shRNA. At 3, 7 and 14 days after transfection, Nell-1 and osteogenesis-related genes (collagen type I, alkaline phosphatase and osteocalcin) were detected by real-time fluorescence quantitative PCR. At 14 days after transfection, ADSCs were detected by alizarin red staining. RESULTS AND CONCLUSION: At 3 days after transfection, there was no significant difference in the Nell-1 mRNA expression between the control and Nell-1 groups (P > 0.05); and the Nell-1 mRNA expression in the control and Nell-1 groups was significantly lower than that in the co-transfection group (P < 0.05). At 7 and 14 days after transfection, the Nell-1 mRNA expression was still lower in the control and Nell-1 groups than in the co-transfection group, and moreover, the expression level in the Nell-1 group was significantly higher than that in the control group (P < 0.05). At 3, 7, and 14 days after transfection, the levels of collagen type I, alkaline phosphatase and osteocalcin mRNA in the co-transfection group were significantly higher than those in the control and Nell-1 groups, and these expression levels were also significantly higher in the Nell-group than in the control group (P < 0.05). At 14 days after transfection, there were more calcium nodules in the co-transfection group than in the Nell-1 group, while no calcium nodules were observed in the control group. The overall results indicate that Nell-1 can enhance the differentiation of ADSCs into osteoblasts, and interference with Noggin gene can upregulate the expression of Nell-1, thereby triggering a synergistic effect on osteogenic differentiation. Figures and Tables | References | Related Articles | Metrics

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A serum-free monolayer method for differentiation of porcine induced pluripotent stem cells into vascular endothelial cells Wei Renyue, Li Xuechun, Li Yan, Yu Yang, Zhang Yu, Liu Zhonghua 2020, 24 (31):  4971-4978.  doi: 10.3969/j.issn.2095-4344.2115 Abstract ( 389 )   PDF (915KB) ( 204 )   Save BACKGROUND: Induced pluripotent stem cells are a cell source with multiple differentiation potentials. Since pig is a suitable model for human cardiovascular and metabolic diseases, induction of porcine pluripotent stem cells into endothelial cells will provide a support for creating a porcine cardiovascular disease model. OBJECTIVE: To establish a serum-free single layer culture method to induce porcine induced pluripotent stem cells into CD31 positive endothelial cells and to identify the obtained endothelial cells. METHODS: We used the serum-free single layer culture method to induce porcine pluripotent stem cells and human embryonic stem cells into endothelial cells under different combinations of CHIR99021, bone morphogenetic protein 4, basic fibroblast growth factor and vascular endothelial growth factor through three stages. The expressions of germ layer markers of the two kinds of stem cells during their differentiation were detected and compared. The endothelial cells obtained after the flow sorting were identified. RESULTS AND CONCLUSION: The endothelial cells differentiated from porcine induced pluripotent stem cells by a single layer culture method showed comparable morphological and functional properties to the immortalized porcine aortic endothelial cells, capable of taking up low-density lipoprotein and forming microvascular-like structure on matrigel. We found that the differentiation efficiency of porcine pluripotent stem cells was higher than that of human embryonic stem cells using the same method, which may be associated with the different expression patterns of lineage-specific markers in the two kinds of cells during the early differentiation. Figures and Tables | References | Related Articles | Metrics

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Exosomes from adipose-derived stem cells can promote cavernous nerve regeneration in rats Gu Yubo, Zhang Dongliang, Yuan Wei, , Song Lujie 2020, 24 (31):  4979-4985.  doi: 10.3969/j.issn.2095-4344.2121 Abstract ( 511 )   PDF (878KB) ( 249 )   Save BACKGROUND: Cavernous nerve is one of the main nerves regulating penile erection, which is easy to be damaged in pelvis surgery and due to trauma and induce erectile dysfunction. The direct effect of stem cell-derived exosome on the regeneration of injured cavernous nerve has not been reported yet. OBJECTIVE: To observe the effect of the exosomes from adipose-derived stem cells on the regeneration of cavernous nerve. METHODS: Three-dimensional culture model of rat nerve tissue in vitro was constructed. After 24 hours of incubation, the experimental group was incubated in the RPMI-1640 medium containing penicillin-streptomycin and exosomes (1015/L) from adipose-derived stem cells. The control group was incubated in the RPMI-1640 medium. The length of axon regeneration at the incisal edge of the cavernous nerve was measured to evaluate the regeneration ability of the cavernous nerve at 2, 5 and 9 days after intervention. RESULTS AND CONCLUSION: Adipose-derived stem cells extracted from rat perirenal adipose tissue adhered with stable morphology and could be induced to differentiate into adipose, bone and cartilage. The exosomes extracted from the passage 3 adipose-derived stem cells showed similar vesicle structure with a diameter near 85 nm. In the three-dimensional culture system, the regeneration ability of the cavernous nerve itself was certain. The axon regeneration length in the experimental group was significantly longer than that in the control group at 2, 5 and 9 days after intervention. The exosomes from adipose-derived stem cells directly act on the cavernous nerve and have a significant promoting effect on nerve regeneration. Figures and Tables | References | Related Articles | Metrics

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Pretreatment of unrelated umbilical cord blood transplantation without antithymocyte globulin for the treatment of acute myeloid leukemia and acute lymphoblastic leukemia: follow-up evaluation of 306 cases Zhang Xuhan, Wang Li, Tang Baolin, Wan Xiang, Yao Wen, Song Kaidi, Sun Zimin 2020, 24 (31):  4986-4993.  doi: 10.3969/j.issn.2095-4344.2141 Abstract ( 510 )   PDF (796KB) ( 48 )   Save BACKGROUND: Umbilical cord blood hematopoietic stem cell transplantation is more and more widely used as a radical treatment for acute leukemia, but its therapeutic effect in different leukemias has not been compared. By comparing the efficacy of diseases, it can guide different patients to choose the transplantation method. OBJECTIVE: To compare and analyze the therapeutic effect of umbilical cord blood hematopoietic stem cell transplantation on acute myeloid leukemia and acute lymphocytic leukemia. METHODS: Clinical data of 306 cases of acute leukemia treated by unrelated umbilical cord blood hematopoietic stem cell transplantation were retrospectively analyzed, including 112 patients with acute myeloid leukemia and 194 with acute lymphoblastic leukemia. All patients received myeloablative conditioning without antithymocyte, and the prevention of graft-versus-host disease was cyclosporine combined with mycophenolate mofetil. RESULTS AND CONCLUSION: (1) Except that the relapse rate after acute lymphoblastic leukemia transplantation was slightly higher than acute myeloid leukemia, the efficacy of the two groups of patients after receiving unrelated umbilical cord blood hematopoietic stem cell transplantation was basically the same. (2) In the group of adolescents and young adults (aged 15-39 years), the rate of neutrophil and platelet implantation in acute myeloid leukemia was faster than in acute lymphoblastic leukemia. Among them, CD34+ cell number and pretreatment program were independent influencing factors for neutrophil implantation, while CD34+ cell number was also an independent influencing factor for platelet implantation. In this age group, the recurrence rate of acute lymphoblastic leukemia patients after transplantation was still higher than that of acute myeloid leukemia, in which chronic graft-versus-host disease was an independent influencing factor. (3) Immune reconstruction testing after transplantation suggests that cord blood CD8+ T cell reconstruction in patients with acute myeloid leukemia was better than in acute lymphoblastic leukemia patients 4 months after transplantation. (4) The above data show that pre-treatment of unrelated cord blood transplantation without antithymocyte globulin has a good effect on acute lymphocytic leukemia and acute myeloid leukemia. Department of Hematology of The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China is qualified for stem cell transplantation. Figures and Tables | References | Related Articles | Metrics

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Effectiveness of unrelated peripheral blood stem cell transplantation in the treatment of severe aplastic anemia Zhang Suping, Sun Ling, Wan Dingming, Cao Weijie, Li Li, Liu Changfeng, Liu Yufeng, Wang Dao, Guo Rong, Jiang Zhongxing, Xie Xinsheng 2020, 24 (31):  4994-5001.  doi: 10.3969/j.issn.2095-4344.2130 Abstract ( 484 )   PDF (780KB) ( 126 )   Save BACKGROUND: The probability of obtaining sibling-matched hematopoietic stem cell transplantation donors for patients with severe aplastic anemia is less than 30%. With the recent improvement of unrelated peripheral blood stem cell transplantation, the efficacy of unrelated peripheral blood stem cell transplantation in the treatment of benign and malignant hematological diseases has reached a result similar to that of sibling-matched hematopoietic stem cell transplantation. OBJECTIVE: To evaluate the efficacy and safety of unrelated peripheral blood stem cell transplantation in the treatment of severe aplastic anemia. METHODS: A retrospective analysis was made in 25 severe aplastic anemia (type I and type II) patients who received unrelated peripheral blood stem cell transplantation from March 2014 to September 2019. All of them were sibling-matched hematopoietic stem cell transplantation-free patients who refused to accept immunosuppressive therapy. The transplantation pretreatment regimen was “fludarabine + cyclophosphamide + rabbit anti-human thymocyte globulin antibody”, and the prevention scheme of graft-versus-host disease was “cyclosporine + mycophenolate mofetil + short-term methotrexate”. The observation indicators included implantation rate, hematopoietic reconstitution time, incidence of graft-versus-host disease, incidence of transplantation-related complications, 5-year estimated overall survival rate and 5-year estimated disease-free survival rate. This study was approved by the Ethics Committee of the First Affiliated Hospital of Zhengzhou University. Patients and their family members signed the informed consent on unrelated peripheral blood stem cell transplantation. RESULTS AND CONCLUSION: (1) Three of the 25 patients died within 28 days after hematopoietic stem cell transplantation, and the implantation could not be evaluated. Of the remaining 22 patients, 21 cases (95.4%) obtained hematopoietic reconstitution and the implantation was successful; 1 case (4.6%) failed. The median time of neutrophil ≥ 0.5×109 L-1 and platelet ≥ 20×109 L-1 was 12 days (9-18 days) and 13 days (10-32 days), respectively. (2) Among the 21 patients with successful implantation, the incidence of acute graft-versus-host disease was 28.6% (6/21), including I-II degree in 3 cases and III–IV degree in 3 cases. There were only 9.5% patients with mild chronic graft-versus-host disease (2/21), and no moderate and severe chronic graft-versus-host disease occurred. (3) During the median follow-up of 396 days (15-1 886 days), 18 cases (72.0%) survived; the 5-year estimated overall survival rate was 71.1%; and the 5-year estimated disease-free survival rate was 65.6%. Totally 7 cases (28.0%) died, of which 3 cases (12.0%) died of infection, 2 cases (8.0%) died of acute intestinal graft-versus-host disease, 1 case (4.0%) died of intracranial hemorrhage, and 1 case (4.0%) died of multiple organ failure. (4) The results show that unrelated peripheral blood stem cell transplantation is a safe and effective treatment for severe aplastic anemia patients without sibling matching donor, and it is a feasible and useful alternative therapy. Figures and Tables | References | Related Articles | Metrics

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Immuoregulatory effects of colorectal cancer cell-derived exosomes on CD8+ T cells Li Zhenxiang, Jiang Xiaokui, Shen Fangfang, Li Shaoshan 2020, 24 (31):  5002-5006.  doi: 10.3969/j.issn.2095-4344.2105 Abstract ( 467 )   PDF (743KB) ( 168 )   Save BACKGROUND: Little has been reported on the effect of exosomes derived from human colorectal cancer cells on the function of immunocytes. OBJECTIVE: To investigate the immunomodulatory effect of exosomes derived from human colorectal cancer cells on CD8+ T cells. METHODS: Exosomes were extracted from Lovo cell lines, and the expression of specific landmarks was detected. Total protein level was detected by BCA assay. Exosomes derived from Lovo cells were co-cultured with human peripheral blood lymphocytes for 96 hours. The proliferation of CD8+ T cells was detected by flow cytometry. The expression of CD38 and HLA-DR markers and levels of interferon-γ and interleukin-2 were detected. RESULTS AND CONCLUSION: The CD63 and CD9 landmarks were extracted. Exosomes derived from Lovo cells could inhibit the expression of CD38+ and HLA-DR indicators of CD8+ T cells and suppress CD8+ T cell proliferation. Additionally, it could decrease the level of interferon-γ secreted by CD8+ T cells. These results indicate that exosomes derived from Lovo cells can inhibit the activation and function of CD8+ T cells. Figures and Tables | References | Related Articles | Metrics

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Effect of glucosamine capsule on cartilage metabolism-related genes in peripheral blood mononuclear cells of patients with knee osteoarthritis Xiao Lei, Hou Ligang 2020, 24 (31):  5007-5012.  doi: 10.3969/j.issn.2095-4344.2108 Abstract ( 386 )   PDF (831KB) ( 49 )   Save BACKGROUND: At present, the research on glucosamine mostly focused on the treatment of knee osteoarthritis, but the research on the effect of glucosamine on cartilage metabolism-related genes in peripheral blood of patients with knee osteoarthritis is limited. OBJECTIVE: To observe the effect of glucosamine capsule on knee osteoarthritis and the expressions of cartilage metabolism-related genes.   METHODS: Totally 90 knee osteoarthritis patients admitted to the Zhengzhou Central Hospital Affiliated to Zhengzhou University from March 2017 to February 2019 were selected, and 40 healthy subjects from the medical examination center in the same period were selected as healthy subjects. The cartilage oligomeric matrix protein (COMP), type II collagen fiber α1 gene (COL2A1), glycoprotein and specific tissue inhibitor (TIMP)-3 gene expression levels in peripheral blood mononuclear cells were detected and compared between healthy subjects and knee osteoarthritis patients before treatment. Knee osteoarthritis patients were divided into routine group and study group by random number table, and they were treated with diclofenac sodium sustained-release tablets, diclofenac sodium sustained-release tablets and glucosamine capsules, respectively. Both groups were treated for 12 weeks. The levels of gene expressions and Lequesne index in peripheral blood mononuclear cells of two groups were compared before and after treatment. During the treatment, the occurrence of adverse reactions was counted in the two groups. RESULTS AND CONCLUSION: (1) The relative expressions of COMP and TIMP-3 in peripheral blood mononuclear cells of knee osteoarthritis patients were higher than those of healthy subjects (P < 0.05), while the mRNA relative expressions of COL2A1 and glycoprotein were lower than those of healthy subjects (P < 0.05). (2) There were no significant difference in gene expression levels in peripheral blood mononuclear cells before treatment between study group and routine group (P > 0.05). The mRNA relative expressions of COMP and TIMP-3 in peripheral blood mononuclear cells of the two groups after treatment were lower than those before treatment, and those of the study group were lower than those of the routine group (P < 0.05). The mRNA relative expressions of COL2A1 and glycoprotein in peripheral blood mononuclear cells of the two groups after treatment were higher than those before treatment, and the mRNA relative expressions of COL2A1 and ACAN in the study group were higher than those in the routine group (P < 0.05). (3) There was no significant difference in Lequesne index between the study group and the routine group before treatment (P > 0.05). Lequesne index in both groups after treatment was lower than that before treatment, and that in the study group was lower than that in the routine group (P < 0.05). (4) There was no significant difference in the incidence of adverse reactions between the study group and the routine group (P > 0.05). (5) Results suggest that glucosamine capsule can effectively improve the clinical symptoms of patients with knee osteoarthritis. It is safe and reliable. It may promote the expression of COL2A1 and glycoprotein genes by inhibiting the expression of COMP and TIMP-3 genes. Figures and Tables | References | Related Articles | Metrics

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Effect of Noggin overexpression in hair follicle dermal sheath cells on the growth of skin and hair follicles Ling Xuejian, Yang Qingchun, Zhang Yanding, Ma Gang 2020, 24 (31):  5013-5017.  doi: 10.3969/j.issn.2095-4344.2091 Abstract ( 556 )   PDF (868KB) ( 90 )   Save BACKGROUND: Noggin is an inhibitory molecule of bone morphogenetic protein (BMP) signaling, which can bind to BMP2/4 to form a complex, thereby blocking BMP signaling and affecting normal development. Some studies have suggested that dermal sheath cells are a kind of dermal stem cells that can self-renew for a long time, and participate in the formation of dermal papilla and dermal sheath during hair follicle regeneration. During the development of hair follicles, Noggin is involved in the induction of hair follicle regeneration, and its loss leads to a decrease in hair follicle number and slows hair follicle growth. However, little is known about the Noggin in the study of hair follicle dermal sheaths. OBJECTIVE: To study the biological function of Noggin protein in the dermal sheath of hair follicles. METHODS: The dermal sheath specific α-SMA-Cre ERT2 tool mice were used to specifically overexpress Noggin protein in the dermal sheath. At 8 and 9 days after birth, αSMA-CreER; pMES-Noggin mice in the experimental group and αSMA-CreER mice in the control group were injected with 4-hydroxytamoxifen. At 21, 23, and 28 days after birth, mouse skin tissues were obtained. Hematoxylin-eosin staining was used to observe hair follicle growth and subcutaneous fat layer thickness, and immunohistochemical staining was used to analyze cell phenotype. RESULTS AND CONCLUSION: Based on the findings of hematoxylin-eosin staining, we found that the growth of hair follicle was not affected, but the growth and development of subcutaneous adipose tissue of hair follicles were seriously impacted, presenting with the subcutaneous fat layer becoming thinned. The immunohistochemical results indicated that the decrease of BMP signaling may result in the thinning of the hair follicle fat layer. This discovery expands the understanding of dermal sheath cells and Noggin protein, and provides an important basis for more accurate understanding of hair follicle regeneration and tissue growth mechanisms. Figures and Tables | References | Related Articles | Metrics

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Wnt5a expression in the regeneration of the inferior segment of rat hair follicle at embryonic and postnatal stages  Cai Bozhi, Chen Xiancai, Ni Na, Lin Weishen, Yuan Yanping, Lin Changmin, Huang Keng 2020, 24 (31):  5018-5022.  doi: 10.3969/j.issn.2095-4344.2136 Abstract ( 547 )   PDF (730KB) ( 88 )   Save BACKGROUND: Wnt5a is the only signaling molecule that is highly expressed in the dermis and closely related to the occurrence of hair follicle dermal components. OBJECTIVE: To preliminarily discuss initiation mechanism of dermal papilla embryogenesis and regeneration in vitro by observing Wnt5a expression of E12.5-E15.5 fetal rat skin and the upper part of tentacle hair follicle during dermal papilla regeneration induced by transplantation. METHODS: The pregnant Sprague-Dawley rats at E12.5, E13.5, E14.5 and E15.5 days were killed under anesthesia. The embryos were taken out to make paraffin sections. The hairy papilla was removed from the tentacle hair follicles of Sprague-Dawley rats, and the hair follicles were removed from the upper segment of the hair follicles and then injected into the skin of the back of Nu/Nu nude mice. The skins of rat embryos at above gestational age and hair follicles at transplanted back positions of the nude mice were taken. Enzymatic immunohistochemical method was used to observe the Wnt5a expression in rat embryonic skin and the upper segment of hair follicle during dermal papilla regeneration induced by transplantation. Hematoxylin-eosin staining was used to observe the regeneration of the inferior segment of hair follicle after transplantation in the upper segment. The animal experiment was approved by the Animal Ethics Committee of Shantou University Medical College. RESULTS AND CONCLUSION: (1) E12.5 Wnt5a was expressed in epidermal cells; E13.5 Wnt5a was expressed in epidermal cells and enhanced; E14.5 Wnt5a was transferred from epidermis to dermis and expressed in dermal cytoplasm; E15.5 Wnt5a was expressed in dermal connective tissue, obviously enhanced, and disappeared in epidermis. (2) From the third day to the ninth day after transplantation of the upper part of the rat hair follicle, the upper fragment of the hair follicle was observed to be growing downward. The complete follicles were re-formed. Wnt 5A was strongly expressed in the regeneration related tissues. (3) The high expression of Wnt5a in the site of dermal papilla formation in this model and dermal papilla embryogenesis suggests that Wnt signaling pathway is closely related to the regeneration of dermal papilla, and Wnt5a may play a very important role.  Figures and Tables | References | Related Articles | Metrics

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Pinellia ternata extract induces apoptosis of leukemia cells by regulating the expression of Bax, Bcl-2 and Caspase-3 proteins Feng Jiakun, Liu Wei, Li Zhengfa, Zhao Xiaochen, Li Zengzheng, Yang Tonghua, Zhao Renbin, Hu Peng, Pei Qiang, Guan Xin 2020, 24 (31):  5023-5029.  doi: 10.3969/j.issn.2095-4344.2120 Abstract ( 642 )   PDF (1015KB) ( 139 )   Save BACKGROUND: Pinellia ternata can be cultivated artificially and is easy to purify. Its extract can inhibit the cell proliferation of chronic myeloid leukemia, acute myelogenous leukemia M3, and T-cell acute lymphoblastic leukemia, but the underlying mechanism remains unclear. OBJECTIVE: To explore the mechanism of action of pinellia ternata extract on apoptosis of three kinds of leukemia cells. METHODS: Chronic myeloid leukemia cell lines (K562), acute myeloid leukemia–M3 cell line (HL-60), acute T lymphocyte leukemia cell line (C8166) were incubated in five kinds of concentration in pinellia ternata extracts in vitro for 24, 48, and 72 hours, respectively. Cell counting kit-8 assay was used to assess the influence of three kinds of leukemia cell proliferation to screen the optimal inhibitory effect of moderate (300 mg/L) and high (500 mg/L) concentration. Flow cytometry was used to determine the incidence of early apoptosis of three kinds of leukemia cells. The mRNA and protein expression levels of Bax, Bcl-2 and Caspase-3 were detected by RT-PCR and western blot assay. RESULTS AND CONCLUSION: Cell counting kit-8 assay results showed that five different concentrations of pinellia extracts could inhibit the proliferation of three kinds of leukemia cells. The inhibition rates of medium concentration (300 mg/L) and high concentration (500 mg/L) were significantly higher than those of low concentration (100 mg/L) and control group (P < 0.01). Flow cytometry (Annexin PE/7AAD and Annexin V/PI) double staining showed that moderate and high concentrations of pinellia extract could induce early apoptosis of three kinds of leukemia cells. RT-PCR results revealed moderate and high concentrations of pinellia extract could down-regulate the mRNA expression level of Bcl-2, and up-regulate the mRNA expression levels of Bax, Bcl-2 , Caspase-3 (P < 0.05, P < 0.01). Western blot assay results showed that the Bcl-2 protein bands of K562 and C8166 cells were significantly weakened after 72 hours of incubation with moderate and high concentrations of pinellia tubulin extract, while no significant changes were observed in HL-60 cells. Bax and caspase-3 protein bands were significantly enhanced. In summary, pinellia ternata extract can effectively inhibit the proliferation of myeloid leukemia and lymphocytic leukemia cells and promote their apoptosis via regulating Bax/Bcl-2 and Caspase-3 protein expression.  Figures and Tables | References | Related Articles | Metrics

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Melatonin promotes Schwann cell migration by activating a typical Wnt/β-catenin signaling pathway Lü Jianwei, Ma Jianxiong, Ma Xinlong 2020, 24 (31):  5030-5037.  doi: 10.3969/j.issn.2095-4344.2138 Abstract ( 480 )   PDF (924KB) ( 182 )   Save BACKGROUND: Melatonin, a neuroendocrine hormone, is mainly secreted by the pineal gland, which can promote the proliferation of Schwann cells and the regeneration of peripheral nerve after injury. OBJECTIVE: To investigate the role of melatonin in the migratory activity of Schwann cells together with the molecular mechanism involved. METHODS: The rodent RSC96 Schwann cell line was cultured in vitro followed by identification. The CCK-8 assay was performed to detect the effect of melatonin on Schwann cells proliferation. The Transwell chamber was adopted to observe the efficacy of melatonin with a series of concentrations, which is 0, 100 nmol/L, 1 μmol/L and 10 μmol/L, on the migratory activity of Schwann cells following 20 hours. The expression of melatonin receptor 1 and 2 in Schwann cells was assessed using qRT-PCR and western blot assay. Transwell cell migration system was conducted to determine the influence of melatonin receptor antagonist luzindole or inhibitor IWR-1 of the canonical Wnt/β-catenin signaling on the enhanced migratory activity exerted by melatonin. Western blot assay was performed to investigate the regulatory effect of melatonin on the activity of Wnt/β-catenin signal cascades.   RESULTS AND CONCLUSION: (1) The cultured RSC96 cells showed the typical morphology of Schwann cells, showing a spindle-like or triangular shape with thin and long processes on the two cell poles. The Schwann cells marker S-100 immunofluorescence staining was positive. (2) Compared with the blank control group, 1 μmol/L and 10 μmol/L melatonin could promote Schwann cell migration dramatically compared to the control group, and displayed a chemoattractive effect, with 1 μmol/L showing the most significant effect. (3) Melatonin receptor 1 was the dominating receptor of melatonin expressed in Schwann cells. The Transwell assay results displayed that the number of migratory cells that melatonin induced was not significantly changed with luzindole added, as compared with treatment with melatonin alone, showing no statistical significance. The number of Schwann cells treated with IWR-1 and melatonin was significantly lower than those treated with melatonin (P < 0.05). (4) Results of qRT-PCR and western blot assay showed that compared with the blank control group, the expression levels of P-Lrp6, LEF-1 and nuclear β-catenin protein in Schwann cells of melatonin group were significantly increased (P < 0.05). (5) The expression levels of LEF-1 and nuclear β-catenin protein in Schwann cells of melatonin group and IWR-1 + melatonin group were significantly higher than those of blank control group and IWR-1 group (P < 0.05), indicating that melatonin can antagonize the inhibition of IWR-1 on Wnt/β-catenin signaling pathway activity to some extent. (6) The above data confirm that melatonin can promote Schwann cell migration by activating Wnt/β-catenin signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Interventional mechanism of Feibi prescription on extracellular matrix transformation in a mouse model of pulmonary fibrosis Cheng Xue, Fang Hong, Zhang Yunke, Wu Yingen 2020, 24 (31):  5038-5043.  doi: 10.3969/j.issn.2095-4344.2112 Abstract ( 503 )   PDF (1128KB) ( 81 )   Save BACKGROUND: In the process of repair, extracellular matrix components first fill up the damaged lung tissue. However, excessive deposition of extracellular matrixes can develop pulmonary interstitial fibrosis. OBJECTIVE: To investigate the effect of Feibi prescription on extracellular matrix transformation in mice with pulmonary fibrosis induced by bleomycin. METHODS: Sixty male C57BL/6 mice were randomly divided into blank control group, model group, pirfenidone group, and three Feibi prescription groups (low-, middle-, and high-dose groups). There were 10 mice in each group. Except for the blank control group, the other five groups were intraperitoneally injected with bleomycin (7.5 mg/kg per day) for 10 continuous days to establish the model of pulmonary fibrosis. On day 1 after modeling, the mice in corresponding drug groups were intragastrically administered with pirfenidone (51.43 mg/kg per day) and Feibi prescription (6.43, 12.86, 25.72 mg/kg per day). Drug administration lasted for 28 days. Then, morphological changes of lung tissue in mice were observed by hematoxylin-eosin staining and Masson staining. The contents of transforming growth factor-β1 and tumor necrosis factor-α in the serum were detected by ELISA, and the expression of a-smooth muscle actin, collagen type I and collagen type III in the lung tissue was detected by western blot assay. RESULTS AND CONCLUSION: The serum levels of transforming growth factor-β1 were significantly increased in the other groups compared with the blank control group (P < 0.01), but the levels were significantly lower in the pirfenidone, middle-dose and high-dose Feibi prescription groups than in the model group (P < 0.05, P < 0.01), and the most decline was in the high-dose Feibi prescription group. Compared with the model group, the expression of tumor necrosis factor-α was significantly reduced in the four drug groups (P < 0.01), but there was no significant difference between the drug groups (P > 0.05). Compared with the blank control group, the expression of α-smooth muscle actin was increased in the other groups to varying degrees (P < 0.05, P < 0.01). The expression of α-smooth muscle actin was significantly lower in the high-dose Feibi prescription group than the low- and middle-dose Feibi prescription groups (P < 0.05). The expression of collagen type I was significantly higher in the model and low-dose Feibi prescription groups than in the control group, whereas it was significantly lower in the middle- and high-dose Feibi prescription groups than in the model group (P < 0.05). Compared with the blank control group, the expression of collagen type III was increased to different extents in the other groups (P < 0.05, P < 0.01). Compared with the low-dose Feibi prescription group, the expression of collagen type III was significantly decreased in the middle-dose Feibi prescription group, high-dose Feibi prescription group and bleomycin group (P < 0.05, P < 0.01). To conclude, Feibi prescription can alleviate pulmonary fibrosis and inhibit the extracellular matrix transformation in bleomycin-induced pulmonary fibrosis mice. The mechanism may be related to down-regulating the levels of transforming growth factor-β1 and tumor necrosis factor-α, and inhibiting the expression of α-smooth muscle actin, collagen type I and collagen type III in the lung tissue. Figures and Tables | References | Related Articles | Metrics

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Th-17 regulatory cytokines promote interleukins-17A and 17F production by neutrophils during asthma Wei Jianghong, Jia Aijun, Ma Libing, Wang Yueling, Qiu Lulu, Xiao Bing 2020, 24 (31):  5044-5051.  doi: 10.3969/j.issn.2095-4344.2114 Abstract ( 416 )   PDF (981KB) ( 35 )   Save BACKGROUND: Th-17 cells and their derived cytokines (interleukins 17, 21, and 22) show an increasing trend in patients with severe asthma, but the specific regulatory mechanism has not yet been determined. OBJECTIVE: To investigate the effect of Th-17 regulatory cytokines on the expression of interleukin 17 in neutrophils in patients with severe asthma. METHODS: There were 28 patients with asthma and 28 healthy controls in the study. Peripheral blood neutrophils isolated from all the subjects were stimulated with interleukins 21, 23, and 6 cytokines and their ability to produce interleukin 17A and 17F was determined relative to healthy controls. Signal transducer and activator of transcription 3 (STAT3) phosphorylation levels were measured in stimulated neutrophils using flow cytometry. The requirement for STAT3 phosphorylation was determined by blocking its activation using a specific chemical inhibitor. The study protocol was reviewed and approved by the Ethics Committee of the Affiliated Hospital of Guilin Medical University, with an approval No. 2017-013. RESULTS AND CONCLUSION: Stimulating asthmatic neutrophils with interleukins 21, 23, and 6 significantly enhanced the production of interleukins-17A and 17F in neutrophils, and increased STAT3 phosphorylation in all the patients compared with the healthy controls. Interestingly, inhibiting STAT3 phosphorylation using a specific chemical inhibitor dramatically blocked the ability of neutrophils to produce interleukin 17, demonstrating that STAT3 activation is the major approach to mediate the expression of interleukin 17 gene. To conclude, neutrophil infiltration in lungs of severe asthmatics may represent an important source of pro-inflammatory interleukins 17A and 17F. STAT3 pathway may be a potential target for regulating neutrophilic inflammation during severe asthma. Figures and Tables | References | Related Articles | Metrics

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Protective effects of sulforaphane in a H2O2 induced oxidative stress model of human hepatocytes Tan Jing, Yi Guodong 2020, 24 (31):  5052-5056.  doi: 10.3969/j.issn.2095-4344.2123 Abstract ( 448 )   PDF (727KB) ( 150 )   Save BACKGROUND: Oxidative stress injury plays an important role in the development of chronic hepatitis, and sulforaphane is a natural product with good antioxidant activity. OBJECTIVE: To investigate the protective effects of sulforaphane on oxidative stress injury of human hepatocytes (LO2) stimulated by H2O2, and to investigate its possible mechanism. METHODS: LO2 cells were cultured in vitro, followed by 24-hour treatment with 100 μg/L H2O2 to induce the oxidative stress model of the cells. The cells were divided into five groups: control group, model group, sulforaphane treatment groups (low-, middle-, high-dose groups: H2O2+10, 20, and 40 μmol/L sulforaphane, respectively). Cell counting kit-8 was used to measure the relative survival rate of cells, and analyze the levels of oxidative stress products in cell homogenate. Western blot assay was used to determine the expression levels of nuclear factor erythroid-2 related factor 2 (Nrf2), GCLC, NQO1 and heme oxygenase-1. The study protocol was in accordance with the related ethic requirements of Central Hospital of Enshi Autonomous Prefecture. RESULTS AND CONCLUSION: H2O2 could induce oxidative stress injury in LO2 cells. Compared with the model group, sulforaphane at various concentrations could significantly induce the proliferation of LO2 cells, inhibit the production of malondialdehyde and nitric oxide, and promote the activity of superoxide dismutase (P < 0.05). Western blot results revealed that sulforaphane could induce the transfer of Nrf2 into the nucleus and significantly increase the expression of GCLC, NQO1, and heme oxygenase-1 proteins (P < 0.05). To conclude, sulforaphane can significantly relieve the oxidative stress injury of LO2 cells induced by H2O2 and its underlying mechanism may be related to the activation of Nrf2 pathway. Figures and Tables | References | Related Articles | Metrics

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Single cell analysis applied to stem cell heterogeneity Gao Shijun, Zhang Haiying, Wang Tan, Chen Li, Peng Yanan, Tang Yunyi, Chen Zhibin, Zhao Zhenqiang 2020, 24 (31):  5057-5063.  doi: 10.3969/j.issn.2095-4344.2096 Abstract ( 456 )   PDF (725KB) ( 171 )   Save BACKGROUND: Single cell analysis of stem cells can reveal the composition and physiological behavior diversity and diversity of various stem cells. As stem cell therapy gradually moves into clinical applications, the accompanying heterogeneity problem is particularly prominent and becomes the key to its development. Therefore, single cell analysis technique for various stem cells has attracted increasing attention. OBJECTIVE: To summarize the single cell isolation and collection technology and progress and application of single cell analysis technology in stem cell heterogeneity research. METHODS: PubMed database was searched for relevant articles published from 1990 to 2019 using the keywords of “(stem cell OR (human pluripotent stem cells) OR (hPSCs)) AND (single cell analysis technology). CNKI and WanFang were retrieved for relevant articles published from 1990 to 2019 using the keywords of “stem cells, single cell analysis” in Chinese. The articles concerning the single cell analysis technique in recent years were reviewed, and finally 57 eligible articles were included for result analysis. RESULTS AND CONCLUSION: Stem cell heterogeneity is the key to blocking the use of stem cells in clinical treatment. Therefore, understanding the heterogeneity of stem cells through single cell analysis is crucial. With the development of single cell high-throughput technology, a new generation of single cell separation and detection technologies will enable people to fully understand single cells. The versatility and control accuracy of the microfluidic device have been significantly improved, and will play a significant role in the analysis of single cells in the stem cells. As a new technology for high-throughput screening, single-cell robots are easier to operate than microfluidics and are highly valued by scientists.  Figures and Tables | References | Related Articles | Metrics

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Physical factors and mechanisms affecting osteogenic ability of bone marrow mesenchymal stem cells Cui Xintao, Zhang Zhenyu 2020, 24 (31):  5064-5070.  doi: 10.3969/j.issn.2095-4344.2132 Abstract ( 451 )   PDF (703KB) ( 53 )   Save BACKGROUND: To accelerate osteogenic differentiation, induce cell proliferation and in turn promote bone formation is considered to be an important research direction in the treatment of osseous diseases. Numerous studies have focused on looking for inducing factors that promote the osteogenic differentiation of bone marrow mesenchymal stem cells. To investigate the optimal conditions for inducing proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells has been an increasing concern. OBJECTIVE: To review and discuss the effects of physical factors related to the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and the possible mechanisms of signal transduction pathways. METHODS: To search for articles on physical factors affecting osteogenic differentiation of bone marrow mesenchymal stem cell in WanFang, CNKI, PubMed, GeenMedical from June 2000 to November 2019, the search terms were “bone marrow mesenchymal stem cells, osteogenesis, differentiation” in Chinese and English, respectively. The old and repeated viewpoints were excluded, and finally 72 literatures were used for analysis. RESULTS AND CONCLUSION: Relevant mechanical factors (such as compressive stress, shear stress, stretch stress, microgravity, low-frequency vibration), electromagnetic field, ultrasonic wave, electrical stimulation, and cold stimulation, have certain effects on the proliferation and osteogenic ability of bone marrow mesenchymal stem cells, which are realized through relevant factors and signaling pathways. However, their effects are influenced by the microenvironment of the transplant site, and some physical factors show a dual effect. Future investigation on the influencing factors with the optimal induction of bone marrow mesenchymal stem cells under different microenvironments as well as the mechanism of these factors will greatly promote the application of stem cells and the development of tissue engineering. References | Related Articles | Metrics

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Factors influencing differentiation of stem cells from the apical papilla into odontoblasts Wang Wenhong, Li Yanjun, Cui Caiyun 2020, 24 (31):  5071-5078.  doi: 10.3969/j.issn.2095-4344.2125 Abstract ( 534 )   PDF (822KB) ( 45 )   Save BACKGROUND: Stem cells from the apical papilla are a kind of odontogenic mesenchymal stem cells that can differentiate into odontoblasts under the action of special inducing factors, further forming the root dentin. Restoration and regeneration of pulp-dentin complex is a new clinical strategy for the treatment of pulp and periapical diseases, especially for the retention of young permanent teeth. Directional induction of stem cells from the apical papilla into odontoblasts has gradually become a hot topic in endodontology. OBJECTIVE: To review the factors related to the differentiation of stem cells from the apical papilla into dontoblasts, in order to provide the research basis for the root development of young permanent teeth and other clinical applications. METHODS: PubMed, VIP, WanFang and China Journal Full-Text databases were searched for relevant literatures published from January 2013 to December 2019 using the keywords of “SCAP, odontogenic differentiation” in English and Chinese, respectively. Finally, 38 articles were retrieved and summarized, and the remaining 18 articles were further explained of the literature content. RESULTS AND CONCLUSION: We systematically reviewed the odontogenic differentiation of stem cells from the apical papilla via relevant signaling pathways under specific induction factors, including cytokines, gene transfection, bioactive materials and scaffolds, chemical factors, and physical factors. Seeking for the ideal conditions for the odontogenic differentiation of stem cells from the apical papilla as well as relevant effects and mechanism is very important for the treatment of pulp and periapical disease and for the study on pulp-dentin complex in tissue engineering field. Figures and Tables | References | Related Articles | Metrics

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Efficacy of exosomes in peripheral nerve injury Yuan Yiming, Wang Yan, Chen Chengcheng, Zhao Mingyue, Pei Fei 2020, 24 (31):  5079-5084.  doi: 10.3969/j.issn.2095-4344.2093 Abstract ( 370 )   PDF (686KB) ( 280 )   Save BACKGROUND: Due to intercellular communication functions and various biological characteristics, exosomes can deal with disease occurrence and development through multiple channels. It has great potential in the treatment of peripheral nerve injury. OBJECTIVE: To summarize the relevant role and mechanism of exosomes in the treatment of peripheral nerve injury, and provide a theoretical basis for clinical treatment of peripheral nerve injury and basic research. METHODS: The articles related to exosomes and nervous system injuries were searched in CNKI and PubMed databases from inception to October 2019. The keywords were “exosome, peripheral nervous injury, miRNA, Schwann cell, inflammation, vessel regeneration” in Chinese and English, respectively. Sixty eligible articles were included for analysis. RESULTS AND CONCLUSION: The structure and function of exosomes are summarized, and it has great advantages as a diagnostic index and treatment plan for various diseases. Exosomes and their main effectors miRNAs, mRNAs and proteins cannot only reflect the pathophysiologic conditions of the derived cells, but also are important ways for intercellular communication and bioactive material transport. In the perspective of the pathways for the treatment of peripheral nerve injury (axon regeneration, regulation of inflammation, angiogenesis), exosomes contribute to the repair of peripheral nerve injury. Figures and Tables | References | Related Articles | Metrics