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    08 March 2022, Volume 26 Issue 7 Previous Issue    Next Issue
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    Bone marrow mesenchymal stem cells regulate ovarian aging in macaques
    Tian Chuan, Zhu Xiangqing, Yang Zailing, Yan Donghai, Li Ye, Wang Yanying, Yang Yukun, He Jie, Lü Guanke, Cai Xuemin, Shu Liping, He Zhixu, Pan Xinghua
    2022, 26 (7):  985-991.  doi: 10.12307/2022.133
    Abstract ( 579 )   PDF (1941KB) ( 65 )   Save
    BACKGROUND: As the core organ of female reproductive system, ovary exerts female reproductive function through ovulation and hormone secretion, and affects the tissues and organs of the whole body. With the acceleration of population aging and a variety of inducements, the number of people with ovarian aging is increasing, but ovarian aging is still in the preliminary research stage, which is still a scientific problem to be solved.
    OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells on the repair of ovary of aging macaques.
    METHODS:  Ten healthy old female monkeys, aged 23-27 years old and weighing 4.5-8.0 kg, were randomly divided into elderly control group (n=4) and elderly treatment group (n=6). The P4 bone marrow mesenchymal stem cells of young macaques were intravenously infused into elderly macaques, once a day, every other day, for three consecutive times. The elderly control group was infused with equal volume of normal saline at the same time. At the 8th month after the infusion of bone marrow mesenchyme stem cells, macaques were euthanized to take ovarian tissues, weighed, and fixed with 40 g/L paraformaldehyde. Hematoxylin-eosin staining was used to observe the structural changes of various levels of follicles for ovary. TUNEL staining was used to calculate the apoptosis rate. Masson staining was applied to analyze the area ratio of collagen. Immunohistochemistry was employed to observe the number of CD34-labeled positive blood vessel.
    RESULTS AND CONCLUSION: (1) The ovarian organ index of the elderly control group was 1.8×10-5, and the ovarian organ index of the elderly treatment group was 6.0×10-5. There was significant difference between the two groups (P < 0.05). (2) Hematoxylin-eosin staining results showed that original, primary, secondary and atresic follicles were seen in the elderly treatment group, but mature follicle was not found; the medulla and stroma were obvious, and there were a few calcium nodules; the elderly control group basically had no follicular structure, only atresic follicle was seen in the local area, which was filled with a lot of fatty tissue. (3) Masson staining results showed that the area ratio of collagen fibers was significantly lower in the elderly treatment group than that in the elderly control group, and significant difference was found between the two groups (P < 0.05). (4) TUNEL staining results showed that the apoptosis rate was lower in the elderly treatment group than that in the elderly control group, and significant difference was found between the two groups (P < 0.05). (5) The results of immunohistochemistry showed that the number of blood vessels in the elderly treatment group was more than that in the elderly control group, and significant difference was found between the two groups (P < 0.05). (6) Results suggest that bone marrow mesenchymal stem cells can promote follicle regeneration, improve ovarian tissue structure, reduce ovarian cell apoptosis, inhibit the progression of ovarian fibrosis, and promote blood vessel regeneration, to delay even reverse ovarian senescence.
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    Exosomes derived from bone marrow mesenchymal stem cells improve the integrity of the blood-spinal cord barrier after spinal cord injury
    Hu Wei, Xie Xingqi, Tu Guanjun
    2022, 26 (7):  992-998.  doi: 10.12307/2022.134
    Abstract ( 758 )   PDF (1995KB) ( 58 )   Save
    BACKGROUND: The studies reveal that stem cells-derived exosomes promote locomotor recovery after spinal cord injury.
    OBJECTIVE: To investigate whether the bone marrow mesenchymal stem cells-derived exosomes promote locomotor recovery via inhibiting the damage of the blood-spinal cord barrier.
    METHODS: Sixty Sprague-Dawley rats were randomly divided into sham operation group, model group, and exosome group (n=20). The rat models of spinal cord injury were established by modified Allen’s method. In the exosome group, 200 μL of bone marrow mesenchymal stem cells-derived exosomes were injected through the tail vein at 30 minutes and 1 day after injury. On the third day after injury, the permeability of the blood-spinal cord barrier was observed. Western blot assay was used to detect the expression of matrix metalloproteinase-9 and claudin-5, Occludin and ZO-1. Gelatin zymography was used to detect the activity of matrix metalloproteinase-9. Immunofluorescence was used to detect the infiltration of neutrophils. Hematoxylin-eosin staining was used to observe the morphological changes of spinal cord injury. At 1, 3, 5, 7, 10, 14, and 21 days after injury, the Basso-Beattie-Bresnahan rating scale was used to evaluate the recovery of motor function.   
    RESULTS AND CONCLUSION: (1) The Basso-Beattie-Bresnahan scores of rats treated with exosome were significantly higher than these spinal cord injury rats at 10, 14, and 21 days after spinal cord injury (P < 0.05). The results of hematoxylin-eosin staining showed that the damage area in the exosome group was significantly reduced compared to the model group (P < 0.05). (2) Exosome treatment significantly reduced the amount of Evans Blue dye extravasation (P < 0.05). Compared with the model group, the expression levels of tight junction protein, including claudin-5, Occludin and ZO-1, were significantly increased in the exosome group (P < 0.05). (3) Exosome inhibited matrix metalloproteinase-9 expression and activity (P < 0.05). (4) Infiltrated myeloperoxidase-positive neutrophils were observed at the lesion site of injured spinal cord at 3 days after spinal cord injury. The exosome treatment significantly inhibited the infiltration of neutrophils (P < 0.05). (5) The results suggested that bone marrow mesenchymal stem cells-derived exosome improved functional recovery after spinal cord injury in part by inhibiting blood-spinal cord barrier disruption and the subsequent infiltration of neutrophils through reducing the degradation of tight junction proteins by inhibiting the expression and activity of matrix metalloproteinase-9.

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    Osteogenic ability of adipose stem cells in diabetic osteoporosis mice
    Gao Yujin, Peng Shuanglin, Ma Zhichao, Lu Shi, Cao Huayue, Wang Lang, Xiao Jingang
    2022, 26 (7):  999-1004.  doi: 10.12307/2022.135
    Abstract ( 687 )   PDF (2321KB) ( 124 )   Save
    BACKGROUND: Long noncoding RNA (LncRNA)-DANCR can regulate multiple biological processes, including bone metabolism. Our gene microarray results showed that LncRNA-DANCR was significantly overexpressed in diabetic osteoporosis mice. 
    OBJECTIVE: To investigate the effect of LncRNA-DANCR on the osteogenic ability of adipose stem cells in diabetic osteoporosis through Wnt/β-catenin signaling pathway.
    METHODS: Diabetic osteoporosis adipose-derived stem cells and control adipose-derived stem cells were isolated and cultured. After 3 days of osteogenic induction, real-time fluorescence quantitative PCR and western blot assay were used to examine the expression of LncRNA-DANCR, Wnt signaling molecules β-catenin and osteogenic transcription factors (RUNX2 and OPN). Diabetic osteoporosis adipose-derived stem cells transfected with lncRNA-DANCR-specific siRNA were designed. After 3 days of osteogenic induction, real-time fluorescence quantitative PCR, western blot assay and alkaline phosphatase staining were used to examine the expression of Wnt signaling molecules β-catenin, osteogenic transcription factors RUNX2 and OPN and osteogenic ability changes.  
    RESULTS AND CONCLUSION: (1) After 3 days of osteogenic induction, the expression level of LncRNA-DANCR in diabetic osteoporosis adipose-derived stem cells group was significantly higher than that in control group. The expression levels of Wnt signaling molecule β-catenin and osteogenic transcription factors RUNX2 and OPN were significantly lower than those in control adipose-derived stem cells. (2) After 3 days of osteogenic induction, β-catenin and RUNX2 and OPN were highly expressed in the siRNA silent group, and the ability of bone formation was enhanced. (3) The results confirmed that LncRNA-DANCR decreased the osteogenic capacity of diabetic osteoporosis adipose-derived stem cells by inhibiting the Wnt/β-catenin signaling pathway. Silencing of LncRNA-DANCR can restore the reduced osteogenic capacity.

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    Hypoxia preconditioning targets and downregulates miR-195 and promotes bone marrow mesenchymal stem cell survival and pro-angiogenic potential by activating MALAT1 
    Hou Jingying, Guo Tianzhu, Yu Menglei, Long Huibao, Wu Hao
    2022, 26 (7):  1005-1011.  doi: 10.12307/2022.136
    Abstract ( 441 )   PDF (2387KB) ( 112 )   Save
    BACKGROUND: Previous studies demonstrated that hypoxia preconditioning promotes bone marrow mesenchymal stem cell survival and angiogenesis. However, specific mechanism remained unclear. Both RNA-MALAT1 (MALAT1) and microRNA-195 (miR-195) are associated with bone marrow mesenchymal stem cell survival and angiogenesis. MALAT1 promotes bone marrow mesenchymal stem cell survival and angiogenesis, whereas miR-195 is a negative regulator of  bone marrow mesenchymal stem cell survival and angiogenesis.
    OBJECTIVE: To investigate whether hypoxia preconditioning could activate MALAT1 to target and downregulate miR-195 to further promote bone marrow mesenchymal stem cell survival and pro-angiogenic potential in vitro. 
    METHODS:  Bone marrow mesenchymal stem cells cultured in vitro were divided into six groups: normoxia (20% O2), hypoxia preconditioning (1% O2), hypoxia preconditioning+si-MALAT1, hypoxia preconditioning+si-MALAT1 negative control, hypoxia preconditioning+miR-195 and hypoxia preconditioning+miR-195 negative control. MALAT1 siRNA and relevant scramble were transfected into the hypoxia preconditioning+si-MALAT1 and hypoxia preconditioning+si-MALAT1 negative control groups respectively prior to hypoxia preconditioning, and miR-195 mimics and relevant negative control were transfected into the hypoxia preconditioning+miR-195 and hypoxia preconditioning+miR-195 negative control groups respectively prior to hypoxia preconditioning. Mesenchymal stem cells in different groups were cultured for 24 hours and cell proliferation, apoptosis and vascularization were evaluated. The expression levels of MALAT1 and miR-195 were detected using qRT-PCR. The potential complementary binding sites of MALAT1 and miR-195 were predicted by RegRNA 2.0. miR-195 mimics and miR-195 mimics negative control were co-transfected into 293T cells with the luciferase reporters containing MALAT1 or MALAT1-mut. Luciferase activity was detected in different groups of cells in order to verify the relationship between MALAT1 and miR-195. 
    RESULTS AND CONCLUSION: (1) Compared with the normoxia group, the hypoxia group presented more rapid growth; cells apoptosis percentage was significantly declined; and number of vascular lumen like structures increased (P < 0.01). The expression of MALAT1 was upregulated, while the level of miR-195 was significantly decreased in the hypoxia group (P < 0.01). (2) Compared with the hypoxia preconditioning and hypoxia preconditioning+si-MALAT1 negative control groups, hypoxia preconditioning+si-MALAT1 group presented much lower growth rate, and cells apoptosis percentage was significantly higher; number of vascular lumen like structures decreased (P < 0.01). The expression of MALAT1 was decreased, while the level of miR-195 was increased (P < 0.01). Compared with the hypoxia preconditioning and hypoxia preconditioning+miR-195 negative control groups, hypoxia preconditioning+miR-195 group also presented much lower growth rate, and cells apoptosis percentage was significantly higher; number of vascular lumen like structures decreased (P < 0.01). The expression of MALAT1 was decreased, while the level of miR-195 was increased (P < 0.01). (3) The dual luciferase reporter assay indicated that compared with miR-195 control and blank control groups, the MALAT1 reporter gene luciferase activity was decreased significantly in miR-195 mimics group, down-regulating about 63% (P < 0.01). However, miR-195 mimics, miR-195 mimics negative control and blank control showed no effect on MALAT1-mut reporter gene (P > 0.05). (4) Hypoxia preconditioning could activate MALAT1 to target and downregulate miR-195 to further promote bone marrow mesenchymal stem cell survival and pro-angiogenic potential.

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    Immunomodulatory effects of deferoxamine and interferon gamma on human dental pulp stem cells
    Zhou Ying, Zhang Huan, Liao Song, Hu Fanqi, Yi Jing, Liu Yubin, Jin Jide
    2022, 26 (7):  1012-1019.  doi: 10.12307/2022.137
    Abstract ( 533 )   PDF (3156KB) ( 60 )   Save
    BACKGROUND: Immune rejection is a great challenge for tissue or organ transplantation, and immunosuppressors often bring serious side effects to patients. The immunosuppressive effect of mesenchymal stem cells brings new hope for the treatment of immune rejection.
    OBJECTIVE: To explore the effect of combined action of deferoxamine and interferon-γ on the immunomodulatory ability of human dental pulp stem cells. 
    METHODS: After pretreatment of human dental pulp stem cells with deferoxamine or interferon-γ or their combination, the proliferation of cells was detected with cell counting kit-8. The mRNA expression of some gene mRNA was examined by Q-PCR. The secretion of some immune regulatory factors was determined using ELISA. Thereafter, pretreated human dental pulp stem cells were co-cultured with mouse spleen lymphocytes, and then the proliferation of lymphocytes was detected by CFSE labeling by flow cytometry. After allogenic skin transplantation, mice were injected with PBS, human dental pulp stem cells or pretreated human dental pulp stem cells into tail vein immediately. The survival time of skin grafts was recorded. The pathology and immunohistochemistry of skin grafts were examined. Moreover, the proportion of Treg cells in spleen of transplanted mice was measured. 
    RESULTS AND CONCLUSION: (1) Deferoxamine or interferon-γ pretreatment inhibited the growth of human dental pulp stem cells, and the inhibition of deferoxamine was more significant than interferon-γ. (2) Pretreatment of deferoxamine combined with interferon-γ significantly increased the gene expression of vascular endothelial growth factor, interleukin 6, transforming growth factor β1, cyclooxygenase 2, human leukocyte antigen G, tumor necrosis factor A inducible protein 6 and interleukin 10, and promote secretion of interleukin 6, transforming growth factor β1 and prostaglandin E2. (3) The results of in vivo experiments confirmed that human dental pulp stem cells pretreated with deferoxamine combined with interferon-γ prolonged the survival time of mouse allograft skin, reduced the infiltration of CD4+T, CD8+T and macrophages at the grafted skin, and increased the proportion of Treg cells in the spleen of recipient mice. (4) It is concluded that deferoxamine combined with interferon-γ pretreatment enhanced the immunomodulatory effect of human dental pulp stem cells, and inhibited the immune rejection of skin allografts.

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    Bushen Huoxue capsule regulates osteogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells via Hedgehog signaling pathway
    Liang Xuezhen, Yang Xi, Li Jiacheng, Luo Di, Xu Bo, Li Gang
    2022, 26 (7):  1020-1026.  doi: 10.12307/2022.138
    Abstract ( 364 )   PDF (3233KB) ( 102 )   Save
    BACKGROUND: Steroid-induced osteonecrosis of femoral head, a common, progressive and refractory orthopedics disease, seriously affected the normal life and work of the patients. The imbalance of osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells was confirmed to involve in the pathogenesis, prevention, and treatment of steroid-induced osteonecrosis of femoral head. However, the mechanism is unclear.  
    OBJECTIVE: To observe the effect of Bushen Huoxue capsule containing serum on osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells treated with glucocorticoids and Hedgehog signal pathway related factors.  
    METHODS:  Bone marrow mesenchymal stem cells were isolated from SD rats and cultured, and intervened with different doses of Bushen Huoxue capsule containing serum and glucocorticoids. CCK-8 assay was used to observe the proliferation of cells. The osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells in different groups were detected by alizarin red staining and oil red “O” staining. The changes of protein expression of osteogenic factors and adipogenic factors and Hedgehog signal pathway related factors of bone marrow mesenchymal stem cells in different groups were analyzed by western blot assay.   
    RESULTS AND CONCLUSION: (1) The Bushen Huoxue capsule containing serum not only could promote the proliferation of normal rat bone marrow mesenchymal stem cells, but also could obviously increase the proliferation activity of bone marrow mesenchymal stem cells treated by the glucocorticoids. (2) After 3 weeks of bone induction, osteogenic induction abilities in the low dose group, middle dose group and high dose group of Bushen Huoxue capsule containing serum were significantly higher than that of the blank control group, especially the middle dose group. (3) After 2 weeks of lipid induction, the positive staining areas revealed by oil red “O” staining in the low dose group, middle dose group and high dose group of Bushen Huoxue capsule containing serum were lower than that of the blank control group, especially the middle dose group. (4) After 3 days of intervention in each group, western blot assay showed that Bushen Huoxue capsule could enhance the protein expression of osteogenic factors (Runx2, Col1a1) and Hedgehog signal pathway related factors (Shh, Gli1) in bone marrow mesenchymal stem cells, and weaken the protein expression of adipogenic factors (PPARγ, C/EBPα) in bone marrow mesenchymal stem cells. (5) The results indicate that the Bushen Huoxue capsule containing serum could promote the osteogenic differentiation of bone marrow mesenchymal stem cells and inhibit the adipocyte differentiation of bone marrow mesenchymal stem cells through the Hedgehog signaling pathway.
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    miR-206 regulates EVI1 gene expression and cell biological behavior in stem cells of small cell lung cancer
    Wang Jifang, Bao Zhen, Qiao Yahong
    2022, 26 (7):  1027-1031.  doi: 10.12307/2022.139
    Abstract ( 340 )   PDF (1394KB) ( 118 )   Save
    BACKGROUND: Through sorting and identification of small cell lung cancer in vitro, the existence of tumor stem cells has been shown to be an important factor to promote the malignant proliferation of tumor cells. Tumorigenesis is closely related to the abnormal expression of various molecules in cells and the regulation of targeted genes. Previous studies have found that EVI1 has the characteristics of proto oncogene and can be up-regulated in lung cancer. miRNA-206 can target the expression of EVI1 gene and protein.
    OBJECTIVE: To investigate the influence of miRNA-206 on cell proliferation and division cycle by targeted regulating the expression of EVI1 transcription gene in stem cells of small cell lung cancer.
    METHODS: Firstly, miRNA-206 transfection experiment was used to verify the targeted regulation of EVI1 gene. The experiment was divided into three groups: miRNA-206 mimics, miR-206 inhibitor, and miR-NC. The expression of EVI1 protein in the stem cells was detected by western blot assay. miRNA-206 lentivirus transfection experiment was used to verify the signal transduction pathway and biological behavior of the cells. The experiment was divided into three groups: pLB-miR-206 group, empty vector group, and control group. The expression levels of miR-206 and EVI1 mRNA in stem cells were detected by RT-PCR. The expression levels of p-Akt and p-JNK protein were detected by western blot assay. The cell proliferation rate was detected by MTT method. Percentage of mitotic and apoptotic cells in cell cycle was detected by flow cytometry.  
    RESULTS AND CONCLUSION: (1) The Evi1 protein level in the miR-206 mimics group was significantly higher than that in the miR-NC group (P < 0.05), and the Evi1 protein level in the miR-206 inhibitor group was significantly lower than that in the miR-NC group (P < 0.05). It was suggested that miR-206 may target the expression of EVI1 gene in stem cells. (2) After transfection with miRNA-206 lentivirus, the expression levels of miR-206 and EVI1 mRNA in pLB-miR-206 group were significantly lower than those in the empty vector and control groups; the protein expression levels of p-Akt and p-JNK decreased; the cell proliferation rate decreased; and the proportion of cell division cycle decreased (P < 0.05). It was found that miR-206 may target to regulate the expression of EVI1 transcription gene and the activation of Akt/JNK signaling pathway in stem cells of small cell lung cancer, which has an impact on cell proliferation and division cycle.
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    Plant-derived basic fibroblast growth factor maintains the growth and differentiation of human embryonic stem cells
    Liu Feng, Peng Yuhuan, Luo Liangping, Wu Benqing
    2022, 26 (7):  1032-1037.  doi: 10.12307/2022.140
    Abstract ( 680 )   PDF (3823KB) ( 41 )   Save
    BACKGROUND: Basic fibroblast growth factor is an important growth factor to maintain the long-term culture of human embryonic stem cells. Providing reliable, non-animal-derived and low-cost basic fibroblast growth factor products is essential for maintaining the growth of human embryonic stem cells and large-scale production.
    OBJECTIVE: To eliminate the potential interference of animal or other bacteria, this study mainly evaluated the effects of plant-derived basic fibroblast growth factor on the growth and differentiation of human embryonic stem cells. 
    METHODS: Human embryonic stem cells line (H9) was used as research materials. Immunofluorescence staining, flow cytometry detection, alkaline phosphatase staining, and RT-PCR were utilized to systematically compare the ability of plant-derived basic fibroblast growth factor to maintain the characteristics of stem cells at different concentrations and its potential to differentiate into ectoderm, mesoderm, endoderm and pancreatic cells. 
    RESULTS AND CONCLUSION: Plant-derived basic fibroblast growth factor (4 μg/L and 0.4 μg/L) and basic fibroblast growth factor (4 μg/L) used in conventional culture system could maintain the growth and differentiation of human embryonic  stem cells, indicating that plant-derived basic fibroblast growth factor can be used as an effective alternative product to support the growth and differentiation of human embryonic stem cells. 
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    Nocardia rubra cell wall skeleton for extemal use improves the viability of adipogenic mesenchymal stem cells and promotes diabetes wound repair
    Wen Dandan, Li Qiang, Shen Caiqi, Ji Zhe, Jin Peisheng
    2022, 26 (7):  1038-1044.  doi: 10.12307/2022.141
    Abstract ( 898 )   PDF (3361KB) ( 131 )   Save
    BACKGROUND: According to the current clinical application of Nocardia rubra cell wall skeleton to the treatment of cervical erosion and its immune effect, considering the possibility of its treatment of chronic wounds. 
    OBJECTIVE: To study the effect of Nocardia rubra cell wall skeleton for extemal use on the proliferation and apoptosis of adipogenic mesenchymal stem cells in vitro, and the impact on the survival rate and skin healing of adipogenic mesenchymal stem cells transplantation in vivo.
    METHODS: Adipogenic mesenchymal stem cells were treated with 10 mg/L Nocardia rubra cell wall skeleton for extemal use. CCK-8 and EDU kits were used to detect the viability and proliferation of adipogenic mesenchymal stem cells. Adipogenic mesenchymal stem cells were treated with 50% sucrose to induce apoptosis, and then with Nocardia rubra cell wall skeleton for extemal use. Afterwards, flow cytometry and TUNEL assay were used to detect the apoptosis. Nocardia rubra cell wall skeleton for extemal use (10 mg/L, 5 mL) was added to passage 3 adipogenic mesenchymal stem cells for 48 hours of culture. The pretreated cells labeled with the fluorescent dye CM-Dil were injected intracutaneously into the wound skin of diabetic mice. The cell survival rate was detected by the LB983 in vivo imaging system. Hematoxylin-eosin staining, Masson staining, and immunofluorescence were used to verify the healing of chronic wounds on day 14.
    RESULTS AND CONCLUSION: Nocardia rubra cell wall skeleton for extemal use could promote the viability and proliferation of adipogenic mesenchymal stem cells, and inhibit their apoptosis. Nocardia rubra cell wall skeleton for extemal use increased the survival rate of adipogenic mesenchymal stem cells under in vivo conditions. The wound healing rate is faster in diabetic mice.
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    Interleukin-8 receptor enhances the migration and adhesion of umbilical cord mesenchymal stem cells to injured endothelium
    Zhu Bingbing, Deng Jianghua, Chen Jingjing, Mu Xiaoling
    2022, 26 (7):  1045-1050.  doi: 10.12307/2022.142
    Abstract ( 470 )   PDF (1819KB) ( 45 )   Save
    BACKGROUND: Endothelial dysfunction is an early predictor of atherosclerosis and cardiovascular disease. Umbilical cord mesenchymal stem cell transplantation provides the possibility for the treatment of vascular injury.
    OBJECTIVE: To explore the effect of overexpression of interleukin-8 receptor A/B on migration of human umbilical cord mesenchymal stem cells. 
    METHODS: Primary cultured human umbilical cord mesenchymal stem cells and human umbilical vein endothelial cells were transfected with adenovirus vector containing interleukin-8 receptor A/B. Confocal immunofluorescence microscopy and western blot assay were used to observe the expression of interleukin-8 receptor A/B in cells. CCK-8 assay, competition test and adhesion test were used to detect the effects of interleukin-8 receptor A/B overexpression on cell proliferation, competitiveness and migration. 
    RESULTS AND CONCLUSION: The fluorescent adenovirus vector overexpressing interleukin-8 receptor A/B has no significant effect on cell proliferation. However, the competition and migration abilities of human umbilical cord mesenchymal stem cells overexpressing interleukin-8 receptor A/B were significantly higher than those of human umbilical vein endothelial cells overexpressing interleukin-8 receptor A/B (P < 0.05). It is concluded that interleukin-8 receptor enhances the migration ability of human umbilical cord mesenchymal stem cells and promotes adhesion to injured endothelium.

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    Effect of naringenin on osteogenic differentiation of human periodontal ligament stem cells
    Luo Xiaoling, Zhang Li, Yang Maohua, Xu Jie, Xu Xiaomei
    2022, 26 (7):  1051-1056.  doi: 10.12307/2022.143
    Abstract ( 660 )   PDF (2523KB) ( 70 )   Save
    BACKGROUND: Naringenin has various physiological activities such as anti-bacterial, anti-inflammatory, anti-fibrosis, and anti-oxidation, and is widely used in the fields of medicine and food. Recent studies have shown that naringenin can effectively promote the osteogenic differentiation of mesenchymal stem cells, but it is unclear whether naringenin regulates the osteogenic differentiation of periodontal ligament stem cells.
    OBJECTIVE: To investigate the effects of naringenin with different concentrations on the osteogenic differentiation of human periodontal ligament stem cells. 
    METHODS: The primary human periodontal ligament stem cells were isolated and cultured. After treatment with naringenin at concentrations of 10, 100 nmol/L, 1, 10, 100 μmol/L and 1 mmol/L for 72 hours, the cytotoxicity of naringenin on periodontal ligament stem cells was measured via CCK-8 assays. The third generation of human periodontal ligament stem cells was cultured with osteogenic medium containing 0, 1, 10 and 100 μmol/L naringenin respectively. Alkaline phosphatase staining and activity detection were performed after 3, 5, and 7 days. Real-time PCR was used to detect the expression of osteogenic factors including Runx2, osteopontin and osteocalcin in human periodontal ligament stem cells after induction for 7 days. Alizarin red staining and quantitative determination of mineralized nodule were performed after induction for 14 days. 
    RESULTS AND CONCLUSION: Naringenin at a concentration of 10 nmol/L-100 μmol/L has no cytotoxicity to human periodontal ligament stem cells, and naringenin at a concentration of 1 mmol/L has obvious cytandotoxicity to human periodontal ligament stem cells. An appropriate concentration of naringenin (1-100 μmol/L) significantly promotes the increase of alkaline phosphatase activity and mineral deposition and upregulates the gene expression of Runx2, osteopontin and osteocalcin. The results showed that naringenin can significantly promote the osteogenic differentiation of human periodontal ligament stem cells, and the ability to promote osteogenic differentiation is strongest at 100 μmol/L. 

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    Huangqin Decoction regulates autophagy to intervene with intestinal acute graft-versus-host disease in mice
    Cui Xing, Sun Xiaoqi, Zheng Wei, Ma Dexin
    2022, 26 (7):  1057-1062.  doi: 10.12307/2022.144
    Abstract ( 614 )   PDF (2075KB) ( 106 )   Save
    BACKGROUND: How to use traditional Chinese medicine to intervene the imbalance of autophagy after intestinal mucosal barrier injury, so as to ultimately intervene the occurrence of gastrointestinal acute graft-versus-host disease, is an urgent problem to be solved after hematopoietic stem cell transplantation.
    OBJECTIVE: To verify the precise mechanism by which Huangqin Decoction interferes with acute intestinal graft-versus-host disease. 
    METHODS:  CB6F1 mice were randomly divided into normal control group, model control group, low-dose Huangqin Decoction group, medium-does Huangqin Decoction group and high-does Huangqin Decoction group, with 16 mice per group. CB6F1 mice in the model control group, low-dose Huangqin Decoction group, medium-does Huangqin Decoction group and high-does Huangqin Decoction group were infused with mononuclear cell suspension (bone marrow cell 8×107 + spleen cell 8×107) obtained from Balb/c mice via caudal vein within 4 hours after 60Co whole body irradiation (radiation dose was 8 Gy). Different concentrations of Huangqin Decoction were given by gavage on the same day after modeling. The rats in the model control group and the normal control group were given the same volume of normal saline by gavage for 15 days. Eight hours after the last gavage, the small intestine tissues of six mice in each group were collected. PCR and western blot assay were used to detect the expression levels of LC3II/I, Beclin1 and P62. The pathological grading of small intestinal mucosa was scored by hematoxylin-eosin staining. The autophagic vesicle structure of small intestinal mucosal epithelial cells was observed by transmission electron microscope. The remaining 10 rats in each group (except the normal control group) were used to observe the clinical grading of acute graft-versus-host disease and record the survival time.   
    RESULTS AND CONCLUSION: (1) After the application of Huangqin Decoction, the survival time of mice was significantly prolonged; the clinical acute graft-versus-host disease score was significantly decreased, and the pathological grading score of small intestinal mucosa was significantly decreased. The score of medium-does Huangqin Decoction group and high-does Huangqin Decoction group was significantly lower than that of model control group, but there was no significant difference between medium-does Huangqin Decoction group and high-does Huangqin Decoction group. (2) The LC3II/I and Beclin1 expression was significantly lower in the model control group than that in the normal control group (P < 0.01), and P62 expression was significantly higher than that in the normal control group (P < 0.01). Huangqin Decoction could promote the recovery of LC3II/I and Beclin1 levels and downregulate p62 levels (P < 0.01). (3) Under transmission electron microscope, the number of autophagic vesicles in the treatment group was significantly higher than that in the model control group, accompanied by the recovery of important organelles such as mitochondria. (4) The results confirm that by interfering autophagy related proteins, Huangqin Decoction can promote the recovery of autophagy in acute graft-versus-host disease, protect intestinal mucosal barrier and reduce intestinal rejection after transplantation and has promise as a new treatment for acute graft-versus-host disease. 

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    Electrophysiological characteristics of cardiomyocytes differentiated from induced pluripotent stem cells
    Xiong Tinglin, Ying Menghui, Zhang Lisha, Zhang Xiaogang, Yang Yan
    2022, 26 (7):  1063-1067.  doi: 10.12307/2022.145
    Abstract ( 572 )   PDF (1305KB) ( 112 )   Save
    BACKGROUND: Transplantation therapy after the directional differentiation of induced pluripotent stem cells can solve the immune elimination and ethical problems, and directional differentiation of induced pluripotent stem cells into cardiomyocytes provides the possibility of cardiomyocyte regeneration research. 
    OBJECTIVE: To detect the electrophysiological function of cardiomyocytes differentiated from induced pluripotent stem cells. 
    METHODS:  By culturing induced pluripotent stem cells in vitro, direct suspension culture and differentiation medium were used to induce induced pluripotent stem cells to form embryoid bodies. Cell growth was observed and formation time, number and contraction rate of cardiomyocytes were recorded. The electrophysiological characteristics of cardiomyocytes were detected by Axon Axopatch 200B single probe ultra-low noise patch clamp amplifier and MED64 multi-electrode array system. The responsiveness of differentiated cardiomyocytes to isoproterenol was observed. Cardiac troponin T expression levels in differentiated cardiomyocytes were detected by immunofluorescence.  
    RESULTS AND CONCLUSION: (1) The growth characteristics of induced pluripotent stem cells were similar to embryonic stem cells, resembling a bird’s nest and growing like a colony. (2) In the differentiation medium culture, induced pluripotent stem cells aggregated to form embryoid bodies of different sizes after 5 days of suspension culture. During the adherent growth of embryoid bodies, up to 10% of the colonies appeared beating cardiomyocytes, and the beating rate on the 20th and 25th days was significantly higher than other time points (P < 0.05). (3) Action potentials of ventricular, atrial and sinoatrial nodular cells were observed in induced pluripotent stem cells differentiated cardiomyocytes. (4) Isoproterenol could increase the spontaneous beat frequency of cardiomyocytes differentiated from induced pluripotent stem cells. (5) Cellular immunofluorescence indicated that cardiomyocytes differentiated from induced pluripotent stem cells were positive for cardiac troponin T. (6) The results confirm that cardiomyocytes differentiated from induced pluripotent stem cells have sinoatrial-like, atrial-like and ventricular-like action potentials, and have adrenaline signal transduction.  

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    Effects of serum containing Wuzang Wenyang Huayu Decoction on phosphorylated-tau protein expression in Alzheimer’s disease cell model
    Wen Xiaoyu, Sun Yuhao, Xia Meng
    2022, 26 (7):  1068-1073.  doi: 10.12307/2022.146
    Abstract ( 579 )   PDF (1817KB) ( 59 )   Save
    BACKGROUND: Various previous studies have shown that Wuzang Wenyang Huayu Decoction has a good therapeutic effect on Alzheimer’s disease, but its pharmacological mechanism has not been fully elucidated.
    OBJECTIVE: To investigate the effect of Wuzang Wenyang Huayu Decoction drug-containing serum on the expression of peroxisome proliferator-activated receptor γ and phosphorylated-tau protein in Alzheimer’s disease cell model. 
    METHODS:  Beta amyloid protein was used to induce primary hippocampal neurons to establish the currently recognized Alzheimer’s disease cell model. Wuzang Wenyang Huayu Decoction drug-containing serum and donepezil hydrochloride drug-containing serum were given for intervention for 72 hours. The dendritic length and branch number of primary neurons cells were detected by immunofluorescence assay. The expression levels of peroxisome proliferator-activated receptor γ and phosphorylated-tau protein were detected by western blot assay.  
    RESULTS AND CONCLUSION: Compared with the control group, the length and branch number of dendritic cells in the model group were significantly decreased (P < 0.05), peroxisome proliferator-activated receptor γ expression decreased, and phosphorylated-tau protein increased (P < 0. 05 or P < 0. 01). Compared with the model group, the dendritic length and branch number of neurons in Wuzang Wenyang Huayu group and donepezil hydrochloride group increased, the expression of PPARγ increased, and the expression of phosphorylated-tau protein significantly decreased (P < 0. 05 or P < 0. 01). Results confirmed that Wuzang Wenyang Huayu Decoction has neuroprotective effect on Alzheimer’s disease cells induced by beta amyloid protein, and its mechanism may be related to upregulation of peroxisome proliferator-activated receptor γ protein and inhibition of phosphorylated-tau protein.
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    Core decompression combined with dental pulp stem cells in the treatment of steroid-associated femoral head necrosis in rabbits
    Wang Xinmin, Liu Fei, Xu Jie, Bai Yuxi, Lü Jian
    2022, 26 (7):  1074-1079.  doi: 10.12307/2022.147
    Abstract ( 469 )   PDF (2707KB) ( 139 )   Save
    BACKGROUND: The dental pulp stem cells have strong proliferation ability and multi differentiation potential ability, which plays an important role in bone and cartilage tissue engineering. Therefore, to study the therapeutic effect of dental pulp stem cells on femoral head necrosis will bring new treatment strategies and hope for patients with femoral head necrosis.
    OBJECTIVE: To observe the therapeutic effect of core decompression combined with dental pulp stem cells on early steroid-associated femoral head necrosis in rabbits.
    METHODS:  Fifty-two adult healthy New Zealand white rabbits were randomly divided into four groups: normal control group, model group, solvent group, and dental pulp stem cell group. Except the normal control group, the model of steroid induced osteonecrosis of femoral head was established in the other three groups. After successful modeling (the 4th week), core decompression was performed on the right side in the solvent group and dental pulp stem cell group. The dental pulp stem cell group was injected with dental pulp stem cells in the decompression hole of the femoral head, and the solvent group was injected with the same volume of sodium chloride injection at the same time. There were three injections in the 4th, 5th and 6th weeks. At the 12th week, indicators such as bone density, bone morphology parameters, empty bone lacuna rate, and bone trabecular area ratio were measured. 
    RESULTS AND CONCLUSION: (1) Imaging examination showed that the treatment effects of solvent group and dental pulp stem cell group were significantly improved compared with the model group. Bone volume fraction, trabecular number, and bone mineral density of femoral head in the dental pulp stem cell group were better than those in the solvent group, and the difference was statistically significant (P < 0.05). (2) Compared with the normal control group, the empty bone lacuna rate and trabecular area ratio of the dental pulp stem cell group were closer; the empty bone lacuna rate of the solvent group was higher than that of the normal control group (P < 0.05); and the trabecular area ratio was lower than that of the normal control group (P < 0.05). The empty bone lacuna rate of the solvent group and dental pulp stem cell group was significantly lower than that of the model group (P < 0.05), and the bone trabecular area ratio was significantly higher than that of the model group (P < 0.05). (3) The results indicate that the treatment of early femoral head necrosis by using core decompression combined with dental pulp mesenchymal stem cells is better than that of simple core decompression.

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    Mechanism by which allicin inhibits proliferation and promotes apoptosis of rat vascular endothelial cells
    Zhang Yujie, Yang Jiandong, Cai Jun, Zhu Shoulei, Tian Yuan
    2022, 26 (7):  1080-1084.  doi: 10.12307/2022.148
    Abstract ( 598 )   PDF (2111KB) ( 42 )   Save
    BACKGROUND: Allicin has anti-fibrosis and anti-angiogenesis effects. Previous studies have confirmed the inhibitory effect of allicin on the proliferation of fibroblasts, but the effect and exact mechanism of allicin on the proliferation and apoptosis of vascular endothelial cells are still unclear.
    OBJECTIVE: To observe the effect of allicin on the morphology, proliferation and apoptosis of vascular endothelial cells and its possible mechanism, and provide a theoretical basis for the next clinical application of allicin.  
    METHODS: Vascular endothelial cells were routinely cultured to logarithmic phase in vitro, and the cells were divided into control group (0 mg/L), low concentration group (25 mg/L), medium concentration group (50 mg/L), and high concentration group (100 mg/L) according to the concentration of allicin in the medium. After 24 hours of intervention, CCK-8 assay was used to detect cell viability. Inverted phase contrast microscope was used to observe cell morphological changes. AO/EB double staining method and AnnexinV-FITC double staining method were used to detect apoptosis rate. PI/RNase method was used to detect cell cycle. Reverse-transcription PCR and western blot assay were used to measure the expression levels of PCNA, Bax and BcL-2 proteins and genes.
    RESULTS AND CONCLUSION: After treated with allicin at concentrations of 0, 25, 50 and 100 mg/L for 24 hours, the viability of vascular endothelial cells significantly decreased in a time-dose-dependent manner, with an IC50 value of 103.27 mg/L at 24 hours. As the concentrations of allicin increased, the apoptosis rate of vascular endothelial cells rose up (P < 0.01); the cell numbers at G1 phase decreased (P < 0.01); and at G2 phase increased (P < 0.01). Compared with control group, the expression of PCNA and Bcl-2 decreased (P < 0.01), while the expression of Bax increased significantly in the allicin group (P < 0.01). The results suggested that allicin inhibited the proliferation of vascular endothelial cells and induced their apoptosis. The action mechanism may be achieved by regulating the expression of PCNA, Bcl-2 and Bax.
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    Systematic evaluation of different therapeutic effects of mesenchymal stem cell transplantation in the treatment of ischemic stroke
    Fang Xiaolei, Leng Jun, Zhang Chen, Liu Huimin, Guo Wen
    2022, 26 (7):  1085-1092.  doi: 10.12307/2022.149
    Abstract ( 560 )   PDF (1421KB) ( 107 )   Save
    OBJECTIVE: To conduct a meta-analysis of the efficacy and safety of mesenchymal stem cells in the treatment of ischemic stroke and to compare the clinical efficacy of different mesenchymal stem cells transplantation pathways. 
    METHODS: PubMed, CNKI, Wanfang data, VIP, Web of Science, and SinoMed were searched, and randomized controlled trials of mesenchymal stem cells in the treatment of ischemic stroke published from January 2005 to March 2021 were collected. The control group received conventional treatment. The stem cell group received mesenchymal stem cell transplantation. The literature quality was assessed using the bias assessment tool recommended by the Cochrane Collaboration. The included primary outcome measures National Institute of Health Stroke Scale score, secondary outcome index modified Rankin Scale score, function independent measure score, Fugl-Meyer assessment score, Barthel index score, and post-treatment adverse outcome were analyzed using RevMan 5.3 and Stata 16.0 software. The network meta-analysis was performed for the National Institute of Health Stroke Scale score of neurological impairment.
    RESULTS: Seventeen randomized controlled trials were included, and the quality of the included articles was generally average. A total of 936 ischemic stroke cases were included. (1) Results of meta-analysis showed that mesenchymal stem cells were superior to conventional treatment group in reducing National Institute of Health Stroke Scale score and modified Rankin Scale score of neurological impairment, improving Fugl-Meyer assessment motor function score, Barthel index score of daily living ability, and functional independence Function independent measure score (P < 0.05). (2) The results of network meta-analysis showed that compared with the conventional treatment group, subarachnoid injection combined with intravenous injection could significantly reduce the National Institute of Health Stroke Scale score (P < 0.05) and promote the recovery of neurological function. SUCRA cumulative probability: Subarachnoid injection combined with intravenous injection (79.0%) > stereotactic implantation (71.3%) > subarachnoid injection (55.8%) > intravenous injection (43.0%) > carotid injection (36.5%) > routine treatment (14.4%). The effect of subarachnoid injection combined with intravenous injection was better than other transplantation methods.
    CONCLUSION: Mesenchymal stem cells can effectively improve neurological function, improve motor function, daily living ability and functional independence in patients with ischemic stroke, and mesenchymal stem cell transplantation is relatively safe. The combination of subarachnoid injection and intravenous injection may be an optimal path to improve the neural function of mesenchymal stem cell transplantation after ischemic stroke.

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    Biological characteristics and immunoregulation of exosomes derived from mesenchymal stem cells
    Guo Jia, Ding Qionghua, Liu Ze, Lü Siyi, Zhou Quancheng, Gao Yuhua, Bai Chunyu
    2022, 26 (7):  1093-1101.  doi: 10.12307/2022.150
    Abstract ( 824 )   PDF (1553KB) ( 417 )   Save
    BACKGROUND: Clarifying the synthesis and regulation of exosomes derived from mesenchymal stem cells is of great significance to the research and application of exosomes in the future.
    OBJECTIVE: To summarize the biological characteristics in exosomes derived from different mesenchymal stem cells and the role of these exosomes in the immune regulation, which hopes to find new breakthroughs in the study of exosomes in the future and provide ideas for cell-free therapy.
    METHODS: The articles were searched on PubMed, CNKI and Wanfang using the keywords of “stem cells, mesenchymal stem cells, exosomes, immunomodulation, inflammatory mediator, biological characteristics of exosomes, biological characteristics of mesenchymal stem cells, regeneration” in Chinese and English. Finally, 105 articles were included for review.   
    RESULTS AND CONCLUSION: Mesenchymal stem cells are a type of pluripotent stem cells that exist widely in various tissues of the body. Mesenchymal stem cells not only have stem cell characteristics, but also play an important role in the immunosuppressive properties. Mesenchymal stem cells had been clinically applied for the treatment of many diseases. Therefore, mesenchymal stem cells were used to manage novel coronavirus pneumonia, and had ideal treatment effect. Exosomes are endosome-derived nanometer-scale vesicles (40–200 nm in diameter). Exosomes derived from mesenchymal stem cells have the same immune regulation function as its maternal cells, including that carrying immunosuppressive factors, promoting macrophages polarization into M2 type, preventing differentiation of pro-inflammatory T cell subsets and inhibiting antigen presentation. 
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    Application of engineered exosomes in bone repair and regeneration
    Wu Weiyue, Guo Xiaodong, Bao Chongyun
    2022, 26 (7):  1102-1106.  doi: 10.12307/2022.151
    Abstract ( 726 )   PDF (1128KB) ( 69 )   Save
    BACKGROUND: Bone defect caused by disease or accident is very common in clinic. The current main treatment method for bone defect repair is autologous bone grafting. Autologous bone grafting may cause complications, such as secondary infections and scars. The other treatments, such as allogeneic bone transplantation, may have risks of immune rejection and disease transmission. Furthermore, safety considerations and the performance of biomaterials and cell-based treatment require further clarification. In recent years, engineered exosomes have shown good application potential in bone repair and bone regeneration, and have become a current research hotspot.
    OBJECTIVE: To review the research progress of engineering exosomes in promoting bone defect repair and bone regeneration.
    METHODS: The PubMed, CNKI and Wanfang databases were searched for relevant articles with the key words of “exosome; engineered exosomes; bone repair; bone regeneration” in English and Chinese, respectively. The search time was from January 1980 to September 2020. Articles that were not related to the purpose of the research and repetitive articles were excluded, and 56 articles that met the criteria were finally included for review. 
    RESULTS AND CONCLUSION: The engineered exosomes can not only promote bone differentiation of osteoblast-related cells and new blood vessel formation, but also target tissues and cells. A large number of experiments have proven that the engineered exosomes for repairing bone defect and promoting bone regeneration is a feasible strategy.

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    Exosomes that deliver specific miRNAs can regulate osteogenesis and promote angiogenesis
    Zhou Hongqin, Wu Dandan, Yang Kun, Liu Qi
    2022, 26 (7):  1107-1112.  doi: 10.12307/2022.152
    Abstract ( 504 )   PDF (1060KB) ( 104 )   Save
    BACKGROUND: Exosomes are nanoscale paracrine vesicles containing a variety of bioactive factors, such as miRNAs. Exosome miRNAs play an important role in intercellular communication. In recent years, more and more studies have focused on whether miRNAs in exosomes promote bone regeneration. 
    OBJECTIVE: To review research status of exosomal miRNAs promoting bone regeneration in the recent years so as to provide a theoretical basis for further research and application in the field of bone regeneration. 
    METHODS: Using “bone regeneration, bone repair, exosomes, miRNA” in Chinese and English as search terms, biomedical literature database, CNKI, and PubMed were retrieved for articles on exosomal miRNA and bone regeneration published from 2010 to 2021. 
    RESULTS AND CONCLUSION: Exosomes derived from different cells can effectively regulate osteogenesis and promote angiogenesis through the delivery of specific miRNAs, which has a broad prospect in bone tissue engineering.

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    Mechanism and application of stem cell-derived exosomes in promoting diabetic wound healing
    Zhang Jinglin, Leng Min, Zhu Boheng, Wang Hong
    2022, 26 (7):  1113-1118.  doi: 10.12307/2022.153
    Abstract ( 621 )   PDF (1117KB) ( 229 )   Save
    BACKGROUND: The exosomes are one of important active components of adult stem cells paracrine, providing the basis for stem cell “non-cellular” treatment. Exosomes from stem cells can play different roles in the wound through different mechanisms to promote the healing of diabetic wounds.
    OBJECTIVE: To summarize the main mechanism and application of stem cell derived exosomes in the treatment of diabetic wounds. 
    METHODS: Computers were used to retrieve English databases such as PubMed database, Web of Science database, FMRS foreign language medical information resource retrieval platform, and Chinese databases such as CNKI and Wanfang Data Knowledge Service Platform. The retrieval scope was from the inception of the database to September 2020. The English and Chinese search terms were “stem cells, exosomes, wound healing, cell proliferation, neovascularization, inflammation, extracellular matrix, therapeutic use”. According to the inclusion and exclusion criteria, 66 articles were included and the results were analyzed. 
    RESULTS AND CONCLUSION: (1) It is summarized that stem cell-derived exosomes can promote diabetic wound healing by regulating inflammation, promoting neovascularization, re-epithelialization and remodeling of extracellular matrix. (2) The application of stem cell-derived exosomes in diabetic wounds was listed. (3) The exosomes from stem cells can avoid the immune response of stem cells in the treatment of diabetic wounds, and the genetic material of stem cells carried by them is more widely regulated and more effective than that from ordinary cells in the treatment of diabetic wounds. (4) At present, the application of stem cell derived exosomes in diabetes wounds still has some problems, such as low purity, high cost of separation, low production efficiency, and the way to reach the target area, loss rate and diffusion efficiency.  
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    Important role of glutathione in stemness and regulation of stem cells
    Huang Chenwei, Fei Yankang, Zhu Mengmei, Li Penghao, Yu Bing
    2022, 26 (7):  1119-1124.  doi: 10.12307/2022.154
    Abstract ( 965 )   PDF (1294KB) ( 93 )   Save
    BACKGROUND: Stem cells possess the capacities of self-renewal and multiple differentiation. Glutathione, an important sulfhydryl compounds, not only participates in the active oxygen metabolism of stem cells, but also plays an important role in self-renewal, proliferation, aging, stemness maintenance and differentiation regulation of stem cells.
    OBJECTIVE: To explore the important role of glutathione in stem cells.
    METHODS: In April 2020, the first author searched the title/abstract with the English keywords of “glutathione or GSH, stem cell”, and searched any field with the Chinese keywords of “glutathione, stem cell”, and searched the related articles included in CNKI, VIP, Wanfang and PubMed databases from 2000 to 2020. A total of 358 Chinese articles and 405 English articles were retrieved. Finally, 86 eligible articles were enrolled for the analysis after deleting the repetitive and non-conforming articles.  
    RESULTS AND CONCLUSION: By reviewing the recent studies on glutathione and stem cells, we found that glutathione played an important role in maintaining stemness, regulating the differentiation of stem cells. In addition, effects of glutathione on cancer stem cells have been verified, which provides more evidence for the future treatment of cancer with reduced glutathione.
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    Theoretical mechanism of traditional Chinese medicine theory on stem cell induced differentiation
    Hui Xiaoshan, Bai Jing, Zhou Siyuan, Wang Jie, Zhang Jinsheng, He Qingyong, Meng Peipei
    2022, 26 (7):  1125-1129.  doi: 10.12307/2022.155
    Abstract ( 605 )   PDF (1181KB) ( 169 )   Save
    BACKGROUND: In recent years, under the guidance of the theory of traditional Chinese medicine, scholars have made certain progress in using traditional Chinese medicine to intervene stem cell induction and differentiation into directional cells and/or tissues, which has gradually become a highlight and hotspot in the field of tissue engineering research. However, there are few articles on traditional Chinese medicine theory and stem cell induced differentiation.
    OBJECTIVE: To discuss the induction and differentiation of stem cells from traditional Chinese medicine theory and summarize its research and progress. 
    METHODS: “Traditional Chinese medicine, theory, induced differentiation, mesenchymal stem cells” were respectively used as search words to retrieve Chinese Journal Full-Text Database, VIP, CBMdisc, Wanfang database, PubMed, and Web of Science from January 1978 to December 2019. With the comprehensive retrieval method of keywords combined with subject words, through the selection of text title and abstract, the author excluded the articles that were not correlated with the research purpose and lacked original and repetitive research, and summarized the 57 articles that finally met the standard.
    RESULTS AND CONCLUSION: The combination of traditional Chinese medicine theory and stem cell differentiation has been applied in the research of induced differentiation of many cells and tissues. In particular, some research achievements have been made in the differentiation and differentiation of stem cells by traditional Chinese medicine therapies and formulas, such as activating blood circulation and removing blood stasis, invigorating qi, nourishing kidney and filling essence. To explore the induction and differentiation of stem cells from traditional Chinese medicine theory can not only open up new directions for further in-depth study of stem cells, but also provide research ideas and guidance for the treatment of related diseases by traditional Chinese medicine, which may become a sign of modernization of traditional Chinese medicine and has great practical significance.
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    Potential of muscle-derived stem cells in peripheral nerve regeneration
    An Weizheng, He Xiao, Ren Shuai, Liu Jianyu
    2022, 26 (7):  1130-1136.  doi: 10.12307/2022.156
    Abstract ( 421 )   PDF (1138KB) ( 128 )   Save
    BACKGROUND: Muscle-derived stem cells have strong self-renewal and multi-directional differentiation potential, which has a wide application prospect in the field of nerve injury repair.
    OBJECTIVE: To review the treatment status of peripheral nerve injury and research progress of muscle-derived stem cells in the treatment of peripheral nerve injury. 
    METHODS: “Muscle-derived stem cell, peripheral nerve injury, tissue engineering, Schwann cell, transplant, regeneration, nerve conduit” were the search words. The articles in PubMed, CNKI and Wanfang databases were cross-searched. After excluding repetitive research and irrelevant articles, 100 articles were finally included for review.  
    RESULTS AND CONCLUSION: Muscle-derived stem cells show good regenerative potential in repairing nerve injury by differentiating into various kinds of support cells needed for nerve regeneration and powerful secretory function.
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    Serial questions about endogenous neural stem cell response in the ependymal zone after spinal cord injury
    Fan Yiming, Liu Fangyu, Zhang Hongyu, Li Shuai, Wang Yansong
    2022, 26 (7):  1137-1142.  doi: 10.12307/2022.157
    Abstract ( 501 )   PDF (1054KB) ( 37 )   Save
    BACKGROUND: Spinal cord injury is a serious injury to the central nervous system, and there is currently no effective treatment for it. Patients with spinal cord injury suffer great pain both physically and mentally. The high cost of treatment and serious sequelae have become the main burden of sick families. With the continuous development of molecular biology research on endogenous neural stem cells, it is found that endogenous neural stem cells have great potential to repair damaged spinal cord conduction bundle, which also brings hope for the reconstruction of functional nerve loop.
    OBJECTIVE: To review the response and regulatory factors of endogenous neural stem cells after spinal cord injury.
    METHODS: PubMed, Web of Science, Wanfang, CNKI China journal full-text database were searched for articles regarding endogenous neural stem cells after spinal cord injury. English and Chinese search terms were “spinal cord injury, endogenous neural stem cell”. According to inclusion and exclusion criteria, 70 articles were included for summary. 
    RESULTS AND CONCLUSION: After spinal cord injury, endogenous neural stem cells in the ependymal region were activated, migrated to the damaged site, differentiated into functional neurons and glial cells, limited the further expansion of the injury, and repaired the damaged neural structure. This process is influenced by a variety of factors released during secondary spinal cord injury. Further research is needed on the origin of endogenous neural stem cells and the mechanisms by which they are activated, as well as how they can be induced to differentiate in a direction more conducive to repair of spinal cord injury.

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    Role of regulatory T cell subsets in liver transplantation and progress in clinical application
    Xuan Juanjuan, Bai Hongtai, Zhang Jixiang, Wang Yaoquan, Chen Guoyong, Wei Sidong
    2022, 26 (7):  1143-1148.  doi: 10.12307/2022.158
    Abstract ( 681 )   PDF (1169KB) ( 106 )   Save
    BACKGROUND: The immune rejection after liver transplantation seriously affects the quality of life of patients. Regulatory T cells play an important role in inhibiting the rejection and maintaining immune tolerance. 
    OBJECTIVE: To review the role of regulatory T cells in liver transplantation and its clinical application, so as to provide ideas for the research and clinical application of regulatory T cells in liver transplantation and other organ transplantation.
    METHODS: The first author searched the relevant articles published from January 1970 to February 2021 in PubMed and CNKI by computer in February 2021. The Chinese and English key words were “regulatory T cells, liver transplantation”. Finally, 49 articles were included and analyzed. 
    RESULTS AND CONCLUSION: (1) Regulatory T cells are a subgroup of T cells that control the autoimmune response in vivo, also known as inhibitory T cells. Regulatory T cells can be divided into naturally occurring natural regulatory T cells and induced adaptive regulatory T cells. Natural regulatory T cells are mainly CD4+CD25+ regulatory T cells, accounting for about 5%-10% of CD4+T cells in peripheral blood and spleen. Regulatory T cells play an important role in inhibiting liver transplantation rejection and maintaining immune tolerance. (2) After liver transplantation, the number and activity of regulatory T cells and the expression of transcription factor forkhead box P3 were increased when the recipient was stimulated by the transplanted liver. (3) The number and activity of regulatory T cells were low in the recipients of rejection after liver transplantation. (4) The number and activity of regulatory T cells were higher in immunotolerant patients after liver transplantation, and regulatory T cells could induce and maintain immune tolerance. (5) Infusion of regulatory T cells into the recipient after liver transplantation can reduce the rejection and prolong the survival time of the transplanted liver. 
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