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    08 March 2024, Volume 28 Issue 7 Previous Issue   
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    Combination of 1% platelet-rich plasma and bone marrow mesenchymal stem cells improves the recovery of peripheral nerve injury
    Feng Ruiqin, Han Na, Zhang Meng, Gu Xinyi, Zhang Fengshi
    2024, 28 (7):  985-992.  doi: 10.12307/2024.109
    Abstract ( 310 )   PDF (9960KB) ( 168 )   Save
    BACKGROUND: Platelet-rich plasma has been shown to enhance the viability and the pro-angiogenesis capacity of mesenchymal stem cells. Extracellular vesicles are one of the key mediators for mesenchymal stem cells to exert their effects, but currently, it is unclear whether platelet-rich plasma affects the functions of extracellular vesicles.
    OBJECTIVE: To investigate the effects of platelet-rich plasma on the function of extracellular vesicles from bone marrow mesenchymal stem cells, verify whether platelet-rich plasma can be used as an adjuvant to enhance the healing effects of bone marrow mesenchymal stem cells on repairing the peripheral nerve injury.
    METHODS: For in vitro study, bone marrow mesenchymal stem cells were cultured under normal conditions and with 1% platelet-rich plasma. The ultracentrifugation was used to extract the extracellular vesicles produced by bone marrow mesenchymal stem cells cultured under normal conditions (EVs-nor) or the condition supplemented with 1% platelet-rich plasma (EVs-prp). Extracellular vesicles were used to incubate with Schwann cells. The EdU assay, western blot assay, qPCR and light microscopy photography were performed to examine the effects of EVs-nor and EVs-prp on Schwann cell reprogramming, which was characterized by cell proliferation, c-Jun expression, reprogramming-associated gene expression and cell morphology. For in vivo study, the model of sciatic nerve injury in rats was established. Bone marrow mesenchymal stem cells were grafted with or without 1% platelet-rich plasma into the injured rat sciatic nerve using a chitin nerve conduit. Eight weeks after the surgery, the recovery was assessed by histological and functional indexes, including regenerated nerve fiber density, gastrocnemius wet weight ratio and sciatic function index.
    RESULTS AND CONCLUSION: (1) Compared with EVs-nor, EVs-prp was stronger in promoting Schwann cell proliferation. The gene expressions of c-Jun and GDNF were significantly upregulated in EVs-prp treated Schwann cells. The morphology of Schwann cells was significantly longer in EVs-prp group than that in EVs-nor group, indicating that EVs-prp had a stronger ability to stimulate Schwann cell reprogramming than EVs-nor. (2) Sciatic nerve injury animal experiment results revealed that grafting mesenchymal stem cells along with platelet-rich plasma into the injured sciatic nerve showed the best recovery compared with grafting mesenchymal stem cells or platelet-rich plasma alone, demonstrated by the significantly improved density of nerve fibers, gastrocnemius wet weight ratio, and sciatic function index. (3) These results suggested that platelet-rich plasma improved the function of bone marrow mesenchymal stem cell-derived extracellular vesicles and could be served as a practical and feasible preparation to synergize with bone marrow mesenchymal stem cells to improve peripheral nerve repair.
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    Overexpression of Sema3A promotes osteogenic differentiation of dental pulp stem cells and MC3T3-E1
    Wang Wen, Zheng Pengpeng, Meng Haohao, Liu Hao, Yuan Changyong
    2024, 28 (7):  993-999.  doi: 10.12307/2024.122
    Abstract ( 173 )   PDF (1562KB) ( 85 )   Save
    BACKGROUND: Sema3A is a power secretory osteoprotective factor. However, studies about Sema3A-modified dental pulp stem cells (Sema3A-DPSCs) are rare.
    OBJECTIVE: To explore the osteogenic differentiation ability of Sema3A-DPSCs and their regulatory effect on the osteogenic differentiation of the pre-osteoblast cell line MC3T3-E1.
    METHODS: First, Sema3A-DPSCs were constructed using a lentivirus infection system carrying the Sema3A gene. Control lentivirus-treated DPSCs (Vector-DPSCs) were used as controls. Sema3A-DPSCs or Vector-DPSCs were co-cultured with proosteoblast line MC3T3-E1 at the ratio of 1∶1 and 1∶3 for 24 hours. Finally, the Sema3A-DPSCs, Vector-DPSCs and their co-cultured cells with MC3T3-E1 were cultured for osteogenic induction and differentiation. Osteogenic gene expression was detected by alkaline phosphatase staining, alizarin red staining and real-time quantitative RT-PCR to evaluate osteogenic differentiation ability.
    RESULTS AND CONCLUSION: (1) Sema3A mRNA and protein expression levels in Sema3A-DPSCs were significantly up-regulated. The level of secreted Sema3A in cell supernatant was up-regulated. (2) Compared with the Vector-DPSCs, mRNA expressions of osteogenic genes alkaline phosphatase, Runt-related transcription factor 2, osteocalcin and Sp7 transcription factors in Sema3A-DPSCs were up-regulated; the activity of alkaline phosphatase was enhanced, and the formation of mineralized nodules increased. (3) There were no obvious differences in proliferation between Sema3A-DPSCs and Vector-DPSCs. (4) Compared with MC3T3-E1/Vector-DPSCs co-culture system, the expression of MC3T3-E1 osteogenic genes was up-regulated, and the total alkaline phosphatase activity was enhanced and more mineralized nodules were formed in the MC3T3-E1/Sema3A-DPSCs co-culture system. (5) The results suggest that overexpression of Sema3A can enhance the osteogenic differentiation of DPSCs. Overexpression of Sema3A in DPSCs can promote osteogenic differentiation of MC3T3-E1 in the DPSCs/MC3T3-E1 co-culture system. 
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    Differentiation of insulin-producing cells from human umbilical cord mesenchymal stem cells infected by MAFA-PDX1 overexpressed lentivirus
    Qiu Xiaoyan, Li Bixin, Li Jingdi, Fan Chuiqin, Ma Lian, Wang Hongwu
    2024, 28 (7):  1000-1006.  doi: 10.12307/2024.104
    Abstract ( 298 )   PDF (3742KB) ( 77 )   Save
    BACKGROUND: Transplantation of stem cell-derived islet β cells has been considered effective for the treatment of type 1 diabetes. Human umbilical cord mesenchymal stem cell is an ideal cellular source, but with a low differentiation efficiency to islet β cells.  
    OBJECTIVE: To explore the possibility of human umbilical cord mesenchymal stem cells modified by MAFA and PDX1 to differentiate into insulin-producing cells.
    METHODS: MAFA-PDX1 lentivirus expression vectors were constructed. The efficiency and potentiality of human umbilical cord mesenchymal stem cells differentiated into insulin-producing cells with three methods were compared by cell morphology, RT-qPCR, and dithizone staining [protocol A: Simple lentivirus group; protocol B: Drug (nicotinamide β-mercaptoethanol) induction followed by lentivirus group; protocol C: lentivirus and drug induction group].  
    RESULTS AND CONCLUSION: (1) Morphological change of cells: Cell morphology was all altered after the induction of three protocols. At day 11, human umbilical cord mesenchymal stem cells induced by protocol B showed the most cell clusters among the three protocols, appearing aggregated islet-like cell clusters. (2) Islet-related gene expression detected by RT-qPCR: Horizontal comparison of the three protocols at the same induction time point showed that the expression levels of MAFA and PDX1 genes were the highest in protocol C on day 5 of induction, and those in protocol B were the highest on day 11 of induction. Human umbilical cord mesenchymal stem cells induced by protocol B had the greatest expression of GCG gene at day 5, INS and GLUT2 genes at day 11. (3) Dithizone staining to identify zinc ions: parts of the post-induced cells were stained brownish red by dithizone on day 11. The partial small island cells were stained brownish red with a darker color (positive expression) in protocol B. (4) It is concluded that the overexpression of MAFA and PDX1 can promote the differentiation of human umbilical cord mesenchymal stem cells into insulin-producing cells. The combination of MAFA-PDX1 gene modification and drug induction is superior to the single gene modification.
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    Efficient strategies for microglia replacement in spinal cord injury models
    Zeng Fanzhuo, Li Yuxin, Sun Jiachen, Gu Xinyang, Wen Shan, Tian He, Mei Xifan
    2024, 28 (7):  1007-1014.  doi: 10.12307/2024.101
    Abstract ( 294 )   PDF (9928KB) ( 47 )   Save
    BACKGROUND: As the incidence of spinal cord injury increases with the years and axon regeneration after spinal cord injury was very difficult. How to promote the recovery from spinal cord injury and improve the transplantation efficiency of stem cells and other therapeutic cells after spinal cord injury has been the focus of clinical and scientific research.  
    OBJECTIVE: To establish the efficient transplantation and replacement of mouse spinal cord microglia in the spinal cord injury model.
    METHODS: CX3CR1 creER-/+::LSL-BDNF-/+-tdTomato mice, CX3CR1+/GFP mice, β-actin GFP mice and C57 BL/6J wild-type mice at 8-10 weeks of age were selected. According to the requirements of the experiment, they were randomly divided into six groups. (1) Sham operation group: eight C57 BL/6J wild-type mice were used when only the lamina was removed without injury. (2) Spinal cord contusion injury group: eight C57 BL/6J wild-type mice were used. (3) Spinal cord crush injury group: eight C57 BL/6J wild-type mice were used. (4) Conjoined symbiotic spinal cord strike injury group: β-actin GFP mice with green fluorescent blood were surgically stitched together with C57 BL/6J wild-type mice, using eight β-actin GFP mice and eight C57 BL/6J wild-type mice. (5) Mr BMT-X Ray group (using PLX5622 to eliminate the spinal microglia and bone marrow transplantation with X-ray radiation): Bone marrow cells from four CX3CR1 creER-/+::LSL-BDNF-/+-tdTomato mice were extracted and transplanted into eight C57 BL/6J wild-type mice for spinal cord injury modeling. (6) Mr BMT-Busulfan group (using PLX5622 to eliminate the spinal microglia and bone marrow transplantation with Busulfan): Bone marrow cells from four CX3CR1+/GFP mice were transplanted into eight C57 BL/6J wild-type mice. The percentage of cell transplantation replacement in this group was observed, and the spinal cord injury model was not established in this group. The sham operation group, spinal cord contusion injury group and spinal cord crush injury group were sampled by perfusion on day 14 after spinal cord injury. The conjoined symbiotic spinal cord strike injury group was sampled by perfusion on day 7 after spinal cord injury. Mr BMT-X Ray group was sampled by perfusion on day 28 after spinal cord injury. Mr BMT-Busulfan group was sampled by perfusion on day 28 after transplantation. The sampling site was a 1.2 cm long spinal cord with the T10 segment as the center. In the Mr BMT-X Ray group and Mr BMT-Busulfan group, additional mouse brain tissue was retained to see if it would lead to brain transplantation and replacement. The number and proportion of transplanted and replaced cells in the damaged area were measured using transgenic mice, symbiosis and immunofluorescence.  
    RESULTS AND CONCLUSION: Compared with the traditional peripheral blood transplantation (9.8%) of mice in the conjoined symbiotic spinal cord strike injury group, the new transplantation methods, Mr BMT-X Ray and Mr BMT-Busulfan, could greatly improve the proportion of spinal microglia transplantation and replacement, which could reach 84.8% and 95.6%, respectively. The difference was significant (P < 0.05). The results showed that Mr BMT-X Ray and Mr BMT-Busulfan could achieve efficient replacement of spinal microglia cells, and could improve the problems of low cell transplantation efficiency, few survival numbers and unclear differentiation of the traditional cell transplantation methods. In addition, Mr BMT-X Ray can only replace the microglia in the spinal cord, while Mr BMT-Busulfan could avoid brain inflammation and injury caused by X-ray radiation transplantation.
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    Role and mechanism of umbilical cord mesenchymal stem cells on polycystic ovary syndrome
    Liu Qiwei, Zhang Junhui, Yang Yuan, Wang Jinjuan
    2024, 28 (7):  1015-1020.  doi: 10.12307/2024.115
    Abstract ( 390 )   PDF (2603KB) ( 53 )   Save
    BACKGROUND: At present, many drugs used in the treatment of polycystic ovary syndrome are super-designated drugs, and the treatment of patients with polycystic ovary syndrome still faces great challenges. Studies have shown that human umbilical cord mesenchymal stem cells can repair ovarian function, but few studies have reported their therapeutic effect on polycystic ovary syndrome.  
    OBJECTIVE: To investigate the therapeutic effect of human umbilical cord mesenchymal stem cells on polycystic ovary syndrome, and to preliminarily explore the correlation between mitochondrial autophagy and the improvement of polycystic ovary syndrome by human umbilical cord mesenchymal stem cells.  
    METHODS: Polycystic ovary syndrome mouse model was established by subcutaneous injection of dehydroepiandrosterone for 20 days into C57BL/6J mice. Human umbilical cord mesenchymal stem cells (2×106) were injected through the caudal vein. After treatment, vaginal secretions were collected for 10 consecutive days to detect the estrus cycle of mice. At 2 weeks after treatment, the levels of sex hormones in the peripheral blood of mice, including luteinizing hormone and follicle-stimulating hormone, were detected by ELISA. Hematoxylin-eosin staining was used to evaluate ovarian histopathology. Finally, mitochondrial autophagy in ovaries was observed by transmission electron microscopy. 
    RESULTS AND CONCLUSION: (1) After human umbilical cord mesenchymal stem cell therapy, follicles at different stages (primitive follicles, primary follicles, and secondary follicles) appeared in the ovary of polycystic ovary syndrome mice, and luteal tissue could be seen, indicating that ovulation function of mice was effectively improved. (2) Polycystic ovary syndrome mice treated with human umbilical cord mesenchymal stem cells had sex hormone levels. (3) Untreated polycystic ovary syndrome mice were found to be in the estrous stage for a long time, lacking estrous interphase and estrous phase, but after human umbilical cord mesenchymal stem cell therapy, the estrous cycle returned to a normal level. (4) After treatment with human umbilical cord mesenchymal stem cells, the mitochondrial autophagy of polycystic ovary syndrome mice was significantly reduced. (5) The results show that human umbilical cord mesenchymal stem cells can effectively improve the symptoms of endocrine disorders and promote ovulation in polycystic ovary syndrome mice, which may be related to the inhibition of mitochondrial autophagy.
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    miR-20a regulates pressure overload-induced cardiac hypertrophy
    Sun Teng, Han Yu, Wang Shuang, Li Jialei, Cao Jimin
    2024, 28 (7):  1021-1028.  doi: 10.12307/2024.107
    Abstract ( 279 )   PDF (2889KB) ( 138 )   Save
    BACKGROUND: Cardiac hypertrophy is an adaptive response of the heart to physiological and pathological stimuli such as pressure overload. It is of compensatory significance in the early stage, but if the stimulation continues, it can cause cardiomyopathy leading to heart failure. MicroRNAs are involved in the regulation of cardiac hypertrophy. However, the role of miR-20a in pressure overload-induced cardiac hypertrophy has not been reported.
    OBJECTIVE: To investigate the role of miR-20a in pressure overload-induced cardiac hypertrophy and the underlying mechanisms.
    METHODS: Transverse aortic constriction was used to induce cardiac hypertrophy in vivo and angiotensin II was used to induce H9c2 cell models of cardiac hypertrophy in vitro. MiR-20a was overexpressed in vivo by intramyocardial injection of miR-20a overexpressing adenovirus and in vitro by transfecting miR-20a mimic into H9c2 cells. Cardiac hypertrophy was assessed by measuring heart weight/body weight ratio, cell surface area, and myocardial fibrosis. The expression levels of atrial natriuretic peptide, brain natriuretic peptide, β-myosin heavy chain and miR-20a were detected by real-time fluorescence quantitative PCR. Mitochondrial fission was detected by MitoTracker. The downstream target genes of miR-20a were predicted by RNAhybrid software.
    RESULTS AND CONCLUSION: (1) The expression level of miR-20a was significantly decreased in both hypertrophic cardiomyocytes and hearts (P < 0.05). (2) At the animal level, overexpression of miR-20a significantly inhibited transverse aortic constriction-induced cardiac hypertrophy, including decreasing the upregulated expression level of hypertrophic marker genes (P < 0.05), reduced the enlarged heart volume, reducing the increased heart weight/body weight ratio (P < 0.01), reducing the increased myocardial cross-sectional area (P < 0.05), and attenuating fibrosis (P < 0.01). (3) At the cellular level, overexpression of miR-20a significantly inhibited angiotensin II-induced cardiomyocyte hypertrophy, including decreasing the upregulated expression levels of atrial natriuretic peptide (P < 0.05), brain natriuretic peptide (P < 0.01) and β-myosin heavy chain (P < 0.05), reducing the increased protein/DNA ratio (P < 0.01), and suppressing the increased cell surface area (P < 0.05). (4) Overexpression of miR-20a significantly inhibited angiotensin II-induced mitochondrial fission (P < 0.05). (5) The results of RNAhybrid software analysis showed that miR-20a and the mRNA 3’ untranslated region of cAMP-dependent protein kinase inhibitor alpha were well complementary and the predicted binding sites were highly conserved. (6) In conclusion, miR-20a is significantly down-regulated in pressure overload-induced cardiac hypertrophy. Overexpression of miR-20a inhibits cardiac hypertrophy at both the cellular level and animal level and attenuates angiotensin II-induced mitochondrial fission.
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    Electroacupuncture intervention on the proliferation and differentiation of hippocampal neurons and oligodendrocytes in Alzheimer’s disease model mice
    Li Longyang, Zhang Songjiang, Zhao Xianmin, Zhou Chunguang, Gao Jianfeng
    2024, 28 (7):  1029-1035.  doi: 10.12307/2024.110
    Abstract ( 234 )   PDF (18796KB) ( 33 )   Save
    BACKGROUND: The effect of electroacupuncture on the proliferation and differentiation of hippocampal oligodendrocytes in model mice with Alzheimer’s disease remains poorly understood while demyelinating reaction related to oligodendrocytes is a common pathological reaction of Alzheimer’s disease.
    OBJECTIVE: To investigate the effects and mechanism of electroacupuncture stimulation of “Baihui” (GV 20), “Fengfu” (GV 16) and bilateral “Shenshu” (BL 23) in Alzheimer’s disease model mice on the proliferation and differentiation of endogenous neural stem cells to neurons and oligodendrocytes.
    METHODS:  Forty 6-week-old SPF APP/PS1 transgenic male Alzheimer’s disease model mice were randomly divided into electroacupuncture group(n=20) and Alzheimer’s disease model group (n=20). Healthy male C57BL/6J mice of the same age were used as normal controls (n=20). The mice in the electroacupuncture group received electroacupuncture at “Baihui” (GV 20), “Fengfu” (GV 16) and bilateral “Shenshu” (BL 23) for 16 weeks (20 minutes/day and one day off a week). After electroacupuncture, Morris water maze was used to detect the changes of learning and memory function. Immunohistochemistry was utilized to detect hippocampal dentate gyrus β-amyloid senile plaques. The expression of BrdU/NeuN and BrdU/GALC in the hippocampal dentate gyrus was detected by immunofluorescence double labeling. Western blot assay was used to detect the expression levels of neuron specific protein Nestin and oligodendrocyte specific protein GALC in the hippocampus. mRNA and protein levels of Notch1 and Hes1 in the hippocampus were detected by real-time fluorescence quantitative PCR and western blot assay. 
    RESULTS AND CONCLUSION: (1) Compared with the normal control group, the ability of learning and memory in the Alzheimer’s disease model group decreased significantly; hippocampal dentate gyrus β-amyloid senile plaques increased significantly (P < 0.01); the expression of GALC and Nestin in the hippocampus decreased significantly (P < 0.01, P < 0.05). (2) Compared with the Alzheimer’s disease model group, the learning and memory ability of the electroacupuncture group was significantly increased; β-amyloid senile plaque in the hippocampal dentate gyrus decreased significantly (P < 0.01). BrdU/NeuN double labeled positive cells in the hippocampal dentate gyrus and Nestin protein expression in the hippocampus increased significantly (P < 0.01, P < 0.05); GALC expression in hippocampus increased significantly (P < 0.01). The mRNA and protein levels of Notch1 in the hippocampus were significantly increased (P < 0.05, P < 0.01). The mRNA and protein levels of Hes1 in the hippocampus decreased significantly (P < 0.05). (3) These findings indicate that electroacupuncture at “Baihui” (GV 20), “Fengfu” (GV 16) and bilateral “Shenshu” (BL 23) of the Alzheimer’s disease model infant mice can promote the proliferation and differentiation of endogenous neural stem cells to neurons and oligodendrocytes, which may be regulated through the Notch1/Hes1 pathway.
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    Riluzole interferes with the activation of NLRP3 inflammasome in microglia of rats with spinal cord injury
    Liu Tao, Zhang Wenkai, Ma Ziqian, Zhang Yan, Chen Xueming
    2024, 28 (7):  1036-1042.  doi: 10.12307/2024.127
    Abstract ( 259 )   PDF (15344KB) ( 36 )   Save
    BACKGROUND: Previous animal studies have shown that riluzole can inhibit neuroinflammatory response after spinal cord injury and promote functional recovery in injured rats, but the study on whether it can regulate the expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in the acute stage is lacking. 
    OBJECTIVE: To observe whether riluzole can reduce microglial pyroptosis and promote functional recovery after spinal cord injury by modulating NLRP3 inflammasome through animal experiments, histological experiments and molecular biology experiments.
    METHODS: Female SD rats were divided into sham operation, model and riluzole groups, with 12 rats in each group. In addition to the sham operation group, T10 spinal cord injury was conducted in rats. The model group was treated with intraperitoneal administration of riluzole with solvent cyclodextrin. The riluzole group was treated with a 4 mg/kg dose of riluzole injection. The effect of riluzole on motor function recovery was assessed using the BBB score and inclined plane test. The recovery of sensory-evoked potential and motor-evoked potential was measured by electrophysiology. Hematoxylin-eosin staining was used to evaluate spinal cord tissue repair. The regulatory effects of riluzole on NLRP3, Caspase-1 and gasdermin D protein expression in spinal cord tissues were detected by western blot assay. ELISA was utilized to detect the expression levels of inflammatory factors interleukin-1β and interleukin-18. The effects of riluzole on the expression of NLRP3, Caspase-1, gasdermin D and interleukin-1β in microglial cells of the injured spinal cord were determined by immunofluorescence staining. 
    RESULTS AND CONCLUSION: (1) At 35 days after spinal cord injury, BBB score and inclined plane test score in the riluzole group were higher than those in the model group (P < 0.05). (2) At 3 days after spinal cord injury, the protein expressions of NLRP3, cleaved Caspase-1, gasdermin D-N (N-terminal domain), interleukin-1β, and interleukin-18 in the spinal cord homogenate of the riluzole group were significantly lower than those of the model group (P < 0.05). (3) At 3 days after spinal cord injury, the fluorescence intensity of NLRP3, Caspase-1, gasdermin D and interleukin-1β in the riluzole group was significantly lower than that in the model group (P < 0.05). (4) At day 35 after spinal cord injury, hematoxylin-eosin staining showed that the area of spinal cord injury in the riluzole group was smaller than that in the model group. Electrophysiological tests showed that the latency periods of sensory-evoked potential and motor-evoked potential in the riluzole group were shorter than those in the model group, and the latency period of wave amplitude in the riluzole group was higher than that in the model group. (5) These results suggest that riluzole can promote the repair of injured spinal cord tissue, promote the repair of nerve conduction function, and further promote the recovery of motor function in rats with spinal cord injury, which may be achieved through the regulation of NLRP3 inflammasome and the reduction of microglial pyroptosis. 
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    Proliferation and metabolic patterns of smooth muscle cells during construction of tissue-engineered blood vessels
    Mei Jingyi, Liu Jiang, Xiao Cong, Liu Peng, Zhou Haohao, Lin Zhanyi
    2024, 28 (7):  1043-1049.  doi: 10.12307/2024.116
    Abstract ( 276 )   PDF (1172KB) ( 61 )   Save
    BACKGROUND: Seed cells are seeded on three-dimensional scaffold materials, and three-dimensional culture in bioreactors is a common in vitro tissue engineering culture method, but the changes in cell proliferation and metabolic patterns in bioengineered blood vessel construction are still unclear.
    OBJECTIVE: To explore the metabolic changes of cells such as oxygen consumption and their causes during the whole process of biological vascular tissue construction by in vitro bioreactor.
    METHODS: The self-built vascular bioreactor system was used as the platform; bovine vascular smooth muscle cells were used as the seed cells, and a conventional CO2 incubator provided the external gas environment for the cultivation process. Seed cells were seeded on a tubular porous polyglycolic acid scaffold material for three-dimensional culture, and the whole process included a one-week resting period and a seven-week pulsating tensile stress stimulation loading period. A non-invasive monitoring system was built, and the optical dissolved oxygen patch method was used to monitor the changes of dissolved oxygen in the culture solution in the reactor, and the glucose consumption and lactic acid production were measured by regular sampling. CCK-8 assay was used to determine the proliferation of smooth muscle cells on polyglycolic acid three-dimensional scaffold materials. Nicotinamide adenine dinucleotide oxidation state and reduction state ratio (NAD+/NADH) was utilized to understand cell proliferation and metabolism in the early stage of culture. RT-qPCR and western blot assay were applied to detect the expression of proliferation-related genes (Ki67) and glycolysis-related genes (GLUT-1, LDHA).
    RESULTS AND CONCLUSION: (1) The dissolved oxygen level in the culture solution was (4.314±0.380) mg/L from the cell injection to the end of the resting period (the first week), and gradually stabilized at (1.960±0.866) mg/L after the tensile stress stimulation (the last seven weeks); the two had significant changes (P < 0.05). (2) The ratio of glucose consumption to lactic acid production in the cell culture medium YL/G increased rapidly after the cells were injected, and the highest value was above 1 on the fifth day, and then slow down to 0.5 (The mean value of YL/G in the resting period was 0.89 and the mean value in the pressurized period was 0.57, P < 0.05). (3) CCK-8 assay results showed that A450 value gradually increased after the cells were injected, and reached the highest value on the fifth day, reaching 3.17, and then slowly decreased. At the same time, it was found that Ki67 mRNA was up-regulated on the third day of culture, and then declined. The expression level of Ki67 protein was higher from the third day to the fifth day. (4) The detection of NAD+/NADH showed that the increase was obvious from the fifth to the seventh day after the injection of cells, and the expression of glycolysis-related genes (GLUT-1 and LDHA) was up-regulated and changed synchronously, and the relative expression was higher in the first five days. (5) The results showed that the tissue-engineered blood vessels were constructed using the vascular bioreactor and the smooth muscle cells in the early stage mainly proliferated and exhibited a metabolic feature of low oxygen consumption. The metabolic characteristics of high oxygen consumption were observed during the pulsatile tensile stress stimulation stage. 
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    Establishment and validation of embryo high-quality prediction models based on the third-day 340 nm absorbance embryo culture
    Zhou Chao, Yu Guangyu, Fan Jiaqi, Yu Chunmei, Wu Min, Chen Shibei
    2024, 28 (7):  1050-1056.  doi: 10.12307/2024.114
    Abstract ( 212 )   PDF (1145KB) ( 126 )   Save
    BACKGROUND: A large number of previous studies have confirmed that a high concentration of metabolites is significantly correlated with embryo quality and clinical outcome, and the theory of silencing embryo development indicates that normally developed embryos maintain a low level of material exchange with the outside world during in vitro culture, while embryos often show abnormal metabolic activity due to stress repair mechanism when DNA damage occurs. 
    OBJECTIVE: To establish and verify an embryo quality prediction model based on the third-day 340 nm absorbance embryo cultures to provide the basis for a more objective and accurate embryo quality assessment. 
    METHODS: 269 patients at the Nanxishan Hospital of Guangxi Zhuang Autonomous Region for in vitro fertilization and embryo transplantation from November 2019 to December 2021 were retrospectively analyzed. Among them, on day 3, 162 cases who had 873 optimal embryos and 214 high-quality blastocysts were included in the high-quality embryo group. On day 3, 107 cases who had 859 non-optimal embryos and 214 non-high-quality blastocysts were included in the non-high-quality embryo group. Lambert-beer law was used to screen out the characteristic wavelength with distinguishing degree between superior and non-superior embryos, analyze its correlation and influence trend with high-quality embryos, and establish the clinical prediction model and validation of absorbance for high-quality and non-high-quality embryos at this wavelength.
    RESULTS AND CONCLUSION: (1) There was a significant difference in absorbance between high-quality and non-high-quality embryos at 340 nm on day 3 (P < 0.001), and a negative correlation was found with the formation of high-quality embryos on day 3 (r=-0.486, P < 0.001). The absorbance of high-quality and non-high-quality blastocyst at 340 nm was significantly different (P < 0.05), and was negatively correlated with the formation of high-quality blastocyst (r=-0.642, P < 0.001). (2) The optimal cut-off value of absorbance at 340 nm between high-quality and non-high-quality embryos on day 3 was 0.235. The area under the curve was 0.799. Sensitivity was 62.9%. Specificity was 78.0%. Accuracy was 70.5%. The optimum cutoff value of high-quality and non-high-quality blastocysts of absorbance at 340 nm was 0.175. The area under the curve was 0.871. Sensitivity was 74.3%. Specificity was 89.1%. Accuracy was 82.2%. (3) Restricted cubic spline curve analysis showed that when the absorbance of the culture medium at 340 nm was greater than 0.221, there was a significant positive trend on the formation of non-high-quality embryos at day 3, and when the absorbance of the culture medium at 340 nm was greater than 0.160, there was a significant positive trend on the formation of non-high-quality blastocysts. (4) The clinical decision curve and clinical influence curve showed that the absorbance of the culture medium at 340 nm had the maximum clinical net benefit for the prediction models of high-quality embryos and high-quality blastocysts on the third day when the valve probability was 0.18-0.95 and 0.16-1.00, respectively, and the ratio of loss to gain within the valve probability range was always less than 1. It is proven that the prediction model has good efficacy in clinical applications. The results of embryo transfer showed that the absorbance of embryo culture medium at 340 nm in non-pregnant patients was significantly higher than that in clinical pregnancy, biochemical pregnancy and early abortion patients (P < 0.05). (5) The high-quality and non-high-quality embryo culture in 340 nm absorbance has a significant difference with correlation. The embryo quality prediction model has a certain clinical value and application effectiveness. The joint embryo morphology evaluation to a certain extent improves the objectivity and accuracy of embryo quality evaluation.
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    Immunomodulatory effect of astragaloside IV on T cells of experimental autoimmune encephalomyelitis mice
    Mu Bingtao, Yu Jingwen, Liu Chunyun, Guo Minfang, Meng Tao, Yang Pengwei, Wei Wenyue, Song Lijuan, Yu Jiezhong, Ma Cungen
    2024, 28 (7):  1057-1062.  doi: 10.12307/2023.766
    Abstract ( 304 )   PDF (5105KB) ( 70 )   Save
    BACKGROUND: In the initial stage of multiple sclerosis, central immune cells activate and release a large number of inflammatory factors, causing white matter demyelination and even involving gray matter neurons. The equilibrium of differentiation between different subsets of CD4+ T cells plays an important role in the progression of experimental autoimmune encephalomyelitis. The previous results of the research group showed that the active ingredient astragalus glycoprotein in astragalus can regulate the immune response in experimental autoimmune encephalomyelitis mice, and whether it has a regulatory effect on the differentiation of T cell subsets has not been determined.  
    OBJECTIVE: To explore the therapeutic effects and immune regulatory mechanisms of astragaloside IV on experimental autoimmune encephalomyelitis mice.
    METHODS: Female C57BL/6 mice were divided into the normal control group, experimental autoimmune encephalomyelitis disease model group, and astragaloside IV treatment group (n=8 per group). Myelin oligodendrocyte glycoprotein peptides 35-55 were used for experimental autoimmune encephalomyelitis model induction in the last two groups. On day 10 to 28 after immunization, the astragaloside IV treatment group was treated with 40 mg/kg per day astragaloside IV intragastrically. Body weight and clinical scores of mice in each group were recorded from the immunization day to the 28th day. On the 28th day after immunization, the mouse spinal cord was taken and made into frozen sections for hematoxylin-eosin staining and Lux fast blue staining to observe pathological changes in the spinal cord. Percentage of splenic T cell subsets was detected using flow cytometry. Western blot assay was used to determine the protein expression of interferon-γ, interleukin-17 and interleukin-6 in the spinal cord. Levels of interferon-γ, interleukin-17, interleukin-6 and interleukin-4 in supernatants of cultured splenocytes were determined by ELISA.  
    RESULTS AND CONCLUSION: (1) Compared with the experimental autoimmune encephalomyelitis disease model group, astragaloside IV could reduce the degree of weight loss in experimental autoimmune encephalomyelitis mice (P < 0.05), ameliorate clinical symptoms (P < 0.05), inhibit the infiltration of inflammatory cells and alleviate myelin loss (P < 0.01, P < 0.05). (2) Compared with the experimental autoimmune encephalomyelitis disease model group, astragaloside IV could inhibit the proportion of CD4+T cell subsets expressing interferon-γ (P < 0.001) and interleukin-17 (P < 0.001), but increase percentages of CD4+ interleukin-10+ (P < 0.001) and CD4+ transforming growth factor-β+ (P < 0.01) T cell subsets. (3) Astragaloside IV could inhibit the expression of interferon-γ (P < 0.05, P < 0.01), interleukin-17 (P < 0.05, P < 0.05), and interleukin-6 (P < 0.05, P < 0.05) in the spinal cord and spleen, and up-regulate the expression of interleukin-4 (P < 0.01) in spleen. (4) These findings confirm that astragaloside IV alleviates clinical symptoms in experimental autoimmune encephalomyelitis mice, which may be related to regulating the splenic T cell subsets, therefore, inhibiting the infiltration of inflammatory cells into the center and reducing the demyelination.
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    Hydroxysafflor yellow A intervenes astrocyte lipocalin 2 expression after cerebral ischemia/reperfusion injury
    Liu Kexin, Song Lijuan, Wu Yige, Han Guangyuan, Miao Zhuyue, Wei Ruheng, Xiao Baoguo, Ma Cungen, Huang Jianjun
    2024, 28 (7):  1063-1069.  doi: 10.12307/2024.119
    Abstract ( 226 )   PDF (1391KB) ( 57 )   Save
    BACKGROUND: Ischemic stroke is a serious threat to human health. After ischemia and hypoxia, astrocyte expresses lipocalin-2 in large amounts to aggravate brain injury, but the specific mechanism is not clear. Hydroxysafflor yellow A has anti-ischemia, anti-oxidation, anti-thrombosis and anti-inflammatory effects. However, whether hydroxysafflor yellow A affects the expression of lipocalin-2 in astrocytes after cerebral ischemia and hypoxia and its mechanism are not clear. 
    OBJECTIVE: To investigate the effect and mechanism of hydroxysafflor yellow A on the expression of lipocalin-2 in astrocytes after cerebral ischemia and reperfusion.
    METHODS: (1) Thirty adult SD rats were randomly divided into three groups: sham operation group, middle cerebral artery occlusion and reperfusion group, and hydroxysafflor yellow A group. The middle cerebral artery occlusion and reperfusion model was established in the latter two groups, and hydroxysafflor yellow A group was intraperitoneally injected with 12 mg/kg hydroxysafflor yellow A after reperfusion. Longa score was used to evaluate the degree of neurological impairment. Infarct volume was determined by TTC staining. JAK2/STAT3 pathway and lipocalin-2 expression were detected by western blot assay and immunofluorescence. Levels of interleukin 1β, interleukin 6 and tumor necrosis factor α were detected by ELISA. (2) Astrocytes were divided into four groups: Normal group, glucose-oxygen deprivation group, hydroxysafflor yellow A group and AG490 group. In the latter three groups, glucose-oxygen deprivation and glucose-oxygen recovery models were established. Astrocytes were treated with 75 μmol/L hydroxysafflor yellow A and 10 μmol/L tyrosine phosphorylation inhibitor AG490 for 8 hours during glucose-oxygen deprivation, respectively. The mechanism of hydroxysafflor yellow A on lipocalin-2 was further verified.  
    RESULTS AND CONCLUSION: (1) Compared with the sham operation group, cerebral infarction was significantly increased in the middle cerebral artery occlusion and reperfusion group, accompanied by aggravated neurological impairment (P < 0.01). Hydroxysafflor yellow A treatment could reduce cerebral infarction volume and improve neurological function (P < 0.01). (2) The expressions of p-JAK2, p-STAT3 and lipocalin-2 in the middle cerebral artery occlusion and reperfusion group were higher than those in the sham operation group (P < 0.01). Hydroxysafflor yellow A treatment reduced the expressions of JAK2, STAT3 and lipocalin-2 (P < 0.01). (3) The expression levels of interleukin 1β, interleukin-6 and tumor necrosis factor α in the middle cerebral artery occlusion and reperfusion group were higher than those in the sham operation group (P < 0.01). Hydroxysafflor yellow A inhibited the expressions of interleukin 1β, interleukin-6 and tumor necrosis factor α (P < 0.01). (4) In vitro, the expressions of p-JAK2, p-STAT3 and lipocalin-2 in the glucose-oxygen deprivation group were significantly higher than those in the normal group (P < 0.01). After adding AG490, the phosphorylation of JAK2 and STAT3 decreased, and the expression of lipocalin-2 was inhibited (P < 0.01). The results suggest that hydroxysafflor yellow A may inhibit the expression of lipocalin-2 in astrocytes after ischemia and hypoxia by regulating the JAK2/STAT3 signaling pathway, thereby reducing brain injury.
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    Expression of N-methyl-D-aspartic acid receptor and endoplasmic reticulum stress related pathway proteins in brain tissue of fluorosis rats
    Yang Chun, Wen Jianxia, Feng Jianglong, Guan Zhizhong, Wei Na
    2024, 28 (7):  1070-1075.  doi: 10.12307/2024.118
    Abstract ( 241 )   PDF (1375KB) ( 115 )   Save
    BACKGROUND: Previous studies have shown that N-methyl-D-aspartic acid receptor (NMDA) receptors are associated with fluorine, but the role in fluoride-induced endoplasmic reticulum stress remains unclear. 
    OBJECTIVE: To observe the changes of excitatory neurotransmitter NMDA receptor and endoplasmic reticulum stress IRE1α-ASK1-JNK pathway protein expression in brain tissue of rats with experimental fluorosis, and to investigate the pathogenesis of neurological injury in fluorosis by giving NMDA receptor inhibitor to SH-SY5Y cells. 
    METHODS: (1) Animal model: 18 1-month-old SD rats were randomly divided into control group (drinking water fluoride content < 0.5 mg/L), low fluoride group (drinking water fluoride content 10.0 mg/L) and high fluoride group (drinking water fluoride content 100.0 mg/L), with 6 rats in each group, half of each sex. After 6 months of fluoride intake, the rats were observed for the occurrence of dental fluorosis, and the 24-hour urinary fluoride content was measured. After anesthesia and euthanasia, the brain tissue of rats was taken to observe the pathological changes. Western blot assay was used to detect NMDA receptors and IRE1α, ASK1 and JNK protein expression in the brain tissue. (2) Cell model: SH-SY5Y cells were cultured in vitro and treated with sodium fluoride at final concentrations of 0.3 mmol/L and 3 mmol/L. The fluoride-stained cells were interfered with 10 μmol/L NMDA receptor antagonists Ifenprodil and MK-801 to observe the relevant protein changes. 
    RESULTS AND CONCLUSION: (1) The incidence of dental fluorosis and urinary fluoride level in rats in the high fluoride group were significantly higher than that in the control and low fluoride groups (P < 0.05). (2) Compared with the control group, the cytoplasm of neuronal cells in the CA3 area of the hippocampus in the low fluoride group was slightly more basophilic, while the neuronal cells in the CA3 area of the high fluoride group were disorganized, with increased basophilicity and some of the nuclei solidified. (3) In rat brain tissue, the expressions of NR2A in the high fluoride group and NR2B in the low fluoride group were significantly higher compared with the control group (P < 0.05), and NR2B, IRE1, ASK1, and p-JNK protein expression levels were increased in the high fluoride group compared with the control and low fluoride groups (P < 0.05). (4) In SH-SY5Y cells, NR1, NR2A and NR2B protein expressions were significantly increased in the high fluoride group compared to the control group (P < 0.05). The protein levels of NR1 and NR2A were significantly reduced in the high fluorine + Ifenprodil group and high fluorine + MK-801 group compared with the high fluorine group (P < 0.05). NR2B protein expression was significantly lower in the high fluorine + Ifenprodil group than that in the high fluorine group (P < 0.05). (5) In SH-SY5Y cells, IRE1, ASK1, and p-JNK protein expression was significantly higher in the high fluoride group compared with the control group (P < 0.05), while ASK1 and p-JNK protein expressions were significantly decreased in the high fluorine + Ifenprodil group and high fluorine + MK-801 group compared with the high fluorine group (P < 0.05). IRE1 protein level was significantly lower in the high fluorine + Ifenprodil group than that in the high fluorine group (P < 0.05). (6) It is concluded that excessive fluorine intake activates NMDA receptors in the central nervous system, causing increased expression of endoplasmic reticulum stress IRE1α, ASK1, and p-JNK proteins, and the use of NMDA receptor inhibitors has a mitigating effect on endoplasmic reticulum stress caused by fluorosis.
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    Extracellular vesicles carrying non-coding RNA regulate the activation of osteoclasts
    Liu Jianhong, Liao Shijie, Li Boxiang, Tang Shengping, Wei Zhendi, Ding Xiaofei
    2024, 28 (7):  1076-1082.  doi: 10.12307/2024.112
    Abstract ( 316 )   PDF (1725KB) ( 72 )   Save
    BACKGROUND: It has been demonstrated that osteoclast activation plays an important role in skeletal system-related diseases. The mechanism of regulation of osteoclast activation by extracellular vesicles carrying non-coding RNA has not been fully elucidated.
    OBJECTIVE: To review and summarize relevant literature in and outside China, and to review the regulation of osteoclast activation by different non-coding RNAs in extracellular vesicles in different diseases, so as to provide a certain direction for subsequent research.
    METHODS: “Non-coding RNA, miRNA, lncRNA, circRNA, snoRNA, osteoclasts, extracellular vesicles, exosome, microparticle, apoptotic bodies” were used as search terms to search the databases of CNKI, WanFang, and VIP. “Extracellular vesicles, exosome, microparticle, apoptotic bodies, non-coding RNA, miRNA, lncRNA, circRNA, snoRNA, osteoclast” were used as search terms to search PubMed. Finally, 71 articles were included.
    RESULTS AND CONCLUSION: (1) The activation of osteoclasts is affected by many factors, among which the specific mechanism of non-coding RNA regulating osteoclast activation is not clear. (2) Extracellular vesicles can be secreted by osteoblasts, bone marrow mesenchymal stem cells, tumor cells and other cells. As a carrier of intercellular communication, extracellular vesicles can carry non-coding RNA to regulate osteoclast activation. (3) In the current studies on the regulation of osteoclast activation by extracellular vesicles carrying non-coding RNA, most of the diseases are osteoporosis, followed by tumor bone metastasis, and most types of non-coding RNA are miRNA. (4) There are relatively few studies on the regulation of extracellular vesicles carrying lncRNA and circRNA and snoRNA on osteoclast activation, and the regulatory mechanism is mainly ceRNA mechanism. (5) In conclusion, an in-depth study of the regulatory mechanism of extracellular vesicles carrying non-coding RNA on osteoclast activation is helpful to find key targets for the treatment of skeletal system-related diseases.
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    Physical factors promote osteogenic differentiation of stem cells
    Wang Shanshan, Shu Qing, Tian Jun
    2024, 28 (7):  1083-1090.  doi: 10.12307/2024.123
    Abstract ( 344 )   PDF (2122KB) ( 92 )   Save
    BACKGROUND: In response to the limitations of traditional repair methods for bone defects, stem cells are widely used in the research of regenerative medicine. Chemical factors are the current research hotspots, but recent studies confirm that the application of physical factors to regulate stem cell differentiation at home and abroad has been intensifying, and physical factors combined with biological scaffolds in bone tissue engineering provide a new idea and method for solving the difficult problem of bone defect repair, with good development prospects.
    OBJECTIVE: To summarize the molecular mechanisms of physical factors such as electromagnetic fields and ultrasound on osteogenic differentiation of stem cells as well as the regulation of signaling pathways and the feasibility of their application in bone tissue engineering. 
    METHODS: A computerized search of the CNKI and PubMed for the last 20 years was conducted. In the title and abstract, we used “stem cell, bone defect, osteogenic differentiation, electromagnetic fields, ultrasound, shock wave, low-level laser therapy, mechanical force, bone tissue engineering” in Chinese and “stem cell, osteoporosis, osteogenic differentiation, electromagnetic fields, ultrasound, bone tissue engineering” in English as search terms. A total of 94 relevant articles were included for review.
    RESULTS AND CONCLUSION: (1) As a non-invasive, non-contact adjuvant therapy, physical factors have a significant impact on bone tissue engineering, and have a positive effect on regulating osteogenic differentiation of stem cells, promoting cell proliferation and viability in bone engineering scaffolds. (2) In addition to activating signaling pathways and osteogenic gene transcription, physical factors can also improve vascularization, increase the volume, area and thickness of bone formed in the stent, promote osseointegration, and improve the success rate of bone scaffolds in regenerating healthy bone tissue. (3) However, the use of physical factors for bone tissue engineering uses different experimental conditions, such as scaffold type, cell type, and intervention conditions, and cannot be directly compared to determine the best parameter settings. There is also a lack of consistency in the effectiveness of these different interventions in promoting fracture healing in clinical use. Therefore, it is necessary to further determine the optimal parameters of physical factors for bone tissue engineering in the future. (4) In general, as an ideal adjuvant therapy, physical factors have great potential in combining with various biomaterials and applying them in bone tissue engineering. 
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    Effect and mechanism of traditional Chinese medicine on inhibiting the aging of mesenchymal stem cells
    Pan Xiaolong, Fan Feiyan, Ying Chunmiao, Liu Feixiang, Zhang Yunke
    2024, 28 (7):  1091-1098.  doi: 10.12307/2023.756
    Abstract ( 417 )   PDF (1725KB) ( 188 )   Save
    BACKGROUND: The aging of mesenchymal stem cells is one of the main causes of aging-related diseases, and seriously affects its clinical application. Traditional Chinese medicine has a good anti-aging effect, and it can inhibit the aging of mesenchymal stem cells to promote its application in tissue engineering and prevent and treat aging-related diseases.
    OBJECTIVE: To review the effect and mechanism of traditional Chinese medicine on inhibiting the aging of mesenchymal stem cells.
    METHODS: We searched CNKI and PubMed for the literature on inhibiting mesenchymal stem cell aging with traditional Chinese medicine from 2012 to 2022. The keywords were “traditional Chinese medicine, mesenchymal stem cells (MSCs), aging” in Chinese and English, respectively. Finally, 92 articles were included for further review.
    RESULTS AND CONCLUSION: (1) We summarized five main mechanisms of the aging of mesenchymal stem cells: DNA damage, telomere shortening, oxidative stress, autophagy disorder and mitochondrial dysfunction. (2) This paper reviewed the phenotypic characteristics of senescent mesenchymal stem cells, including increases in cell volume, decreases in proliferation and multi-directional differentiation, increases in β-galactosidase activity, and activation of p21 and p16 pathways, and so on. (3) We summarized the main mechanisms of Chinese medicine inhibiting the senescence of mesenchymal stem cells at present, including inhibiting the activation of the Wnt pathway, inhibiting the production of mitochondrial reactive oxygen species, promoting the silencing of information regulator factor 2 homolog 1, phosphatidylinositol 3-kinase/protein kinase B, adenosine 5’-monophosphate-activated protein kinase and nuclear factor E2 related factor 2 pathway activation, and promoting the expression of telomerase reverse transcriptase. (4) At present, bone marrow mesenchymal stem cells are the most widely studied in the research of traditional Chinese medicine to inhibit the aging of bone marrow mesenchymal stem cells, and the effect is better. (5) Zuogui Wan, Bushen Tiaogan Formula, resveratrol, Astragalus membranaceus and other traditional Chinese medicines can prevent and treat osteoporosis by promoting the proliferation and osteogenic differentiation of aging mesenchymal stem cells. However, the mechanism of Chinese medicine in improving the paracrine function of mesenchymal stem cells and preventing other aging-related diseases by inhibiting the aging of mesenchymal stem cells needs to be further explored. 
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    Bibliometric analysis of researches on liver organoids
    Xu Canli, He Wenxing, Wang Lei, Wu Fangting, Wang Jiahui, Duan Xuelin, Zhao Tiejian, Zhao Bin, Zheng Yang
    2024, 28 (7):  1099-1104.  doi: 10.12307/2024.134
    Abstract ( 376 )   PDF (30880KB) ( 124 )   Save
    BACKGROUND: In recent years, the development of liver organoids has made it a hot spot in the field of international liver disease research, but there is still no article on the bibliometric analysis of liver organoids.
    OBJECTIVE: To explore the hot trends in liver organoids in the last 20 years based on bibliometrics and visualization analysis. 
    METHODS: We searched the articles about liver organoids in the Web of Science Core Collection from January 1, 2002 to November 12, 2022. Origin, Office, and CiteSpace software were used for bibliometrics and visualization analysis. We statistically analyzed the number of annually published articles, countries, institutions, authors, journals, and keywords of the articles by generating charts. 
    RESULTS AND CONCLUSION: The number of articles, citation frequency, institutions and personnel involved in the research about liver organoids showed an overall upward trend in the last 20 years, indicating that the field was growing rapidly and attention was increasing. The USA had published the most papers and had the strongest influence in this field. Although it had invested a lot of time and energy, the number of papers published by a single research institution in the USA was not the highest among many research institutions. China was second only to the USA in the number of publications, with the Chinese Academy of Sciences and Fudan University leading the list. Utrecht University in the Netherlands was the institution with the most publications. Clevers H was the author with the highest number of articles. The article with the highest co-citation frequency was “Long-term culture of genome-stable bipotent stem cells from adult human liver”. The main fields of study for liver organoids were Molecular Science, Biology, and Immunology. The most frequently occurring keywords were stem cell, in vitro, and culture. The research hotspots in the liver organoids field were mainly focused on in vitro stem cell three-dimensional culture, differentiation and gene expression. 
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    Mechanism of satellite cell regulation and its role in ecological niche signaling during skeletal muscle regeneration
    Kong Jianda, Mu Yujing, Zhu Lei, Li Zhilin, Chen Shijuan
    2024, 28 (7):  1105-1111.  doi: 10.12307/2023.745
    Abstract ( 364 )   PDF (2004KB) ( 75 )   Save
    BACKGROUND: Satellite cells are a specific population of adult stem cells contained in skeletal muscle that promote the regenerative reconstruction of injured skeletal muscle, but their specific mechanisms are not well established.  
    OBJECTIVE: To review the regulatory role of satellite cells during skeletal muscle regeneration and the mechanism of interaction between satellite cells and their ecological niche signals, aiming to provide new research ideas and perspectives based on the summary of existing knowledge.
    METHODS: Web of Science, PubMed, CNKI, WanFang, and VIP databases were searched for literature published between January 2002 and June 2022. English search terms were “muscle, skeletal muscle, muscle injury, stem cells, satellite cells, muscle repair”. Chinese search terms were “skeletal muscle, skeletal muscle regeneration, skeletal muscle reconstruction, satellite cells, ecological niche”. The 66 included papers were organized and analyzed.  
    RESULTS AND CONCLUSION: (1) Satellite cells exist in skeletal muscle and contribute to both the formation of new muscle fibers after injury and the effective growth of existing adult muscle fibers. (2) After the activation of quiescent satellite cells in satellite cells, the steps of satellite cell proliferation, differentiation and fusion to form muscle fibers during skeletal muscle regeneration are influenced by their intrinsic regulatory effects of different mechanisms. (3) Satellite cells can interact with myofibers, extracellular matrix, skeletal muscle junctions, fibroblast progenitor cells, immune cells and endothelial cells in the ecological niche signal to promote satellite cell activation, proliferation and differentiation to achieve effective skeletal muscle regeneration. (4) Possible breakthroughs in future research include: the division pattern of satellite cells in the body; the mechanisms regulating satellite cell transfer; the specific timing of satellite cell differentiation or self-renewal in vivo; and the interaction mechanisms between satellite cells and skeletal muscle junctions. (5) This review may provide some theoretical reference values for the field of injury reconstruction of skeletal muscle and its innovation.
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    Role of autophagy in hair regeneration
    Huang Yuxin, Liang Wenzi, Chen Xiuwen, Ni Na, Zhao Yinglin, Lin Changmin
    2024, 28 (7):  1112-1117.  doi: 10.12307/2024.102
    Abstract ( 423 )   PDF (1842KB) ( 62 )   Save
    BACKGROUND: Autophagy has become a rapidly developing research hotspot in the biomedical fields. Many researchers are actively exploring the molecular regulatory mechanism of autophagy in a variety of diseases. However, the role of autophagy in hair growth is still unknown. 
    OBJECTIVE: To review the current research progress and application value of autophagy in hair growth and regeneration, to understand the role of autophagy in hair growth, to explore the pathogenesis of autophagy in pathological hair loss, and to provide new ideas for the study of drugs for hair loss.
    METHODS: Using “hair follicle growth, hair growth, hair regeneration, autophagy associated proteins, autophagy activity, autophagy associated genes, autophagy” as Chinese search terms and “hair growth, hair follicle, hair regeneration, autophagy” as English search terms, PubMed and CNKI databases were searched. The research progress on autophagy, hair growth and the role of autophagy in hair growth in and outside China in recent years was reviewed and summarized. Articles incompatible with the subject content of the paper were excluded. Finally, 78 articles were included for the result analysis.
    RESULTS AND CONCLUSION: (1) Autophagy is a normal metabolic process in eukaryotes with complex molecular mechanisms and functional properties, which is beneficial to cell survival and cell death. (2) Alopecia-related diseases are associated with changes in autophagy activity, which can regulate hair growth cycle. Knockout or overexpression of autophagy-related genes can change the state of hair growth. Multiple autophagy related signaling pathways have been found to be related to hair follicle growth. Activators or inhibitors of autophagy can be used to treat or prevent hair loss.
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    Artificial exosomes in treatment of myocardial infarction: current status and prospects
    Liu Hanfeng, Wang Jingjing, Yu Yunsheng
    2024, 28 (7):  1118-1123.  doi: 10.12307/2024.111
    Abstract ( 394 )   PDF (1798KB) ( 101 )   Save
    BACKGROUND: Myocardial infarction is one of the most serious cardiovascular diseases at present, and the existing clinical treatment options such as thrombolytic therapy, percutaneous coronary intervention and coronary artery bypass grafting cannot fully restore the myocardial damage caused by ischemia. Stem cell-derived exosomes for the treatment of myocardial infarction have been a hot research topic in recent years, but the low yield of natural-derived exosomes, the difficulty and time consuming nature of obtaining them, and the poor homing effect have limited their clinical application. In this context, the construction of artificial exosomes as an alternative to natural exosomes has become an effective strategy to solve the above problems.  
    OBJECTIVE: To expound the research status of artificial exosomes in the treatment of myocardial infarction, and classify them into two design modes: semi-artificial and full-artificial, and discuss the research progress and problems of the two modes, finally, make the evaluation and prospect of its clinical application in the future.
    METHODS: PubMed and CNKI were searched for relevant articles with “artificial exosomes, myocardial infarction, engineering” in Chinese, and “artificial exosome, hybrid exosome, myocardial infarction, nanoparticle, drug delivery system” in English. The focus of the search was from January 2017 to December 2022, and some of the classic forward literature was included. A preliminary selection was conducted through reading titles and abstracts. Repetitive studies, low-quality journals and irrelevant articles were excluded. Finally, 73 articles were included for review.  
    RESULTS AND CONCLUSION: (1) By semi-artificially modifying exosomes, whether it is the modification of targeting peptides, hybridization of biofilms or the assistance of magnetic substances, traditional exosome therapies with insufficient targeting and low retention rate and easy to be cleared by the reticuloendothelial system have improved the efficiency of traditional exosome therapy for myocardial infarction. However, these strategies have problems such as unclear modification efficiency, medical ethics, and biotoxicity. (2) Fully artificial bionic exosomes have a higher degree of design freedom compared to exosome modification, which can solve the problems of high extraction and storage difficulties of exosomes of natural origin and limitations of large-scale production; however, this artificial exosome strategy still lacks reliable preclinical data support and biosafety testing, and has not yet formed a standardized process required for large-scale production; therefore, before applying to the clinic, the artificial exosome solution as an alternative to natural exosomes still needs continuous in-depth research by researchers.
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    Role and mechanism of noncoding RNA in diabetic peripheral neuropathy
    Liu Tao, He Zhijun, Li Jinpeng, Song Yuan, Yao Xingzhang, Chen Wen, Li Yan, Bai Bihui
    2024, 28 (7):  1124-1129.  doi: 10.12307/2023.780
    Abstract ( 253 )   PDF (1618KB) ( 90 )   Save
    BACKGROUND: Persistent hyperglycemia has been identified as promoting neurovascular dysfunction, leading to irreversible endothelial dysfunction, increased neuronal apoptosis, oxidative stress and inflammation. These changes in combination or alone lead to microvascular and macrovascular lesions as well as progressive neuropathy. Noncoding RNAs may provide a new strategy for understanding the etiology, pathogenesis and treatment of the disease.  
    OBJECTIVE: To review the role and mechanism of noncoding RNAs in the occurrence and development of diabetic peripheral neuropathy by reviewing relevant literature at home and abroad, in order to provide new ideas and approaches for noncoding RNAs in the prevention, diagnosis and treatment of diabetes neuropathy. 
    METHODS: CNKI and PubMed were retrieved for relevant literature published from database inception to 2022. The key words were “noncoding RNA; lncRNA; miRNA; diabetes peripheral neuropathy; expression profile” in Chinese and English, respectively. The retrieved documents were summarized and analyzed, and 61 articles were finally selected for further review. 
    RESULTS AND CONCLUSION: (1) Noncoding RNA plays a key role in the pathophysiological process of diabetic peripheral neuropathy. Among the most widely studied regulatory noncoding RNA species, there are long noncoding RNAs, circular RNAs and microRNAs. (2) Through the regulation of noncoding RNAs, the activation or inhibition of related cell pathways, inflammatory genes and downstream-related cytokines will inhibit cell apoptosis, improve inflammation, and thus change the expression of target genes to participate in the process of diabetic neuralgia. (3) Although many microRNAs and long noncoding RNAs have been found to participate in diabetic peripheral neuropathy, the mechanisms of many noncoding RNAs are unclear, and the same noncoding RNAs may play different roles in different modes. Therefore, it is necessary to further study their action modes in disease etiology and pathology, thereby clarifying their role in the pathogenesis of diabetic peripheral neuropathy. However, the criteria for evaluating noncoding RNA activity have not yet been established, and further research is needed on which specific noncoding RNAs play a dominant regulatory role. (4) MicroRNAs, long noncoding RNAs and their target genes can regulate progressive neuropathy, which are expected to become new targets for the clinical prevention and treatment of diabetic peripheral neuropathy and new biomarkers for the development and prognosis of diabetic peripheral neuropathy.
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    Potential value of canonical and non-canonical roles of connexin 43 in disease treatment
    Zhuge Xiaoxuan, Li Ce, Bao Guangjie, Kang Hong
    2024, 28 (7):  1130-1136.  doi: 10.12307/2023.905
    Abstract ( 299 )   PDF (1638KB) ( 45 )   Save
    BACKGROUND: Connexin 43 (Cx43), which is thought to be engaged in the gap junction process and build the structural groundwork for the development of direct material signaling channels between cells, is crucial for maintaining the homeostatic balance of tissue metabolism. Recent research, however, has revealed fresh information about its distinct hemichannel function and highlighted the significance of its subcellular localization and self-fragmentation for cellular physiological activities and pathological processes.
    OBJECTIVE: To systematically summarize the molecular characteristics and expression of Cx43 in a variety of cells, concentrate on the pathological and physiological roles of channel-dependent Cx43 and channel-independent Cx43, and investigate the potential value in disease treatment by reviewing the pertinent literature in the database.
    METHODS: The Chinese and English keywords were “gap junction, connexin 43 (Cx43), hemichannel, channel-dependent Cx43, channel-independent Cx43, extracellular vesicles (EVs), mitochondria, GJA1-20k”, which were searched in PubMed and CNKI. Finally, 81 articles were selected for review. 

    RESULTS AND CONCLUSION: (1) The canonical role of Cx43 is to form a gap junction channel. Channel-dependent Cx43 has primarily involved in disease physiopathological processes by directly constituting gap junction channels, but full attention should be paid to the issue of its structural and functional integrity. Adhesion is a crucial characteristic of gap junctions, which are strongly associated with barrier-like diseases. (2) The non-canonical role of Cx43 is non-gap junction channel-dependent effect. In addition to being localized at the plasma membrane, inner mitochondrial membrane, extracellular vesicle surface, and other structures, Cx43 hexamer has also been found to play a role in positive pro-inflammatory mechanisms, mitochondrial functional metabolism, and targeted uptake of extracellular vesicles in inflammatory diseases. Selective shortened segments control mitochondrial homeostasis by encouraging the polymerization of peri-mitochondrial actin and are involved in the targeted translocation of full-length Cx43 to intracellular structural domains. (3) The development of targeted medicines and the solving of issues like the mechanism of seed cell transformation in tissue engineering-based therapies are both made possible by these two categories of impacts. The interactions of various types of Cx43, however, are frequently not fully taken into account in some of the existing original studies, which confuses the overall characteristics and skews the results. (4) It is necessary to systematically frame the physiological characteristics of Cx43 in different forms and its potential mechanisms in various diseases, so as to provide a reference for the exploration of the Cx43 integrity mechanism and the diagnosis and treatment of multiple diseases.

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    Exosomes derived from mesenchymal stem cells in treatment of animals with acute liver failure: a meta-analysis
    Ma Shuwei, He Sheng, Han Bing, Zhang Liaoyun
    2024, 28 (7):  1137-1142.  doi: 10.12307/2024.113
    Abstract ( 268 )   PDF (16312KB) ( 28 )   Save
    OBJECTIVE: To evaluate the efficacy of exosomes derived from mesenchymal stem cells on animal models of acute liver failure.
    METHODS: PubMed, Web of Science, Embase, The Cochrane Library, CBM, CNKI, WanFang, and VIP databases were retrieved from inception to January 16, 2023. A series of animal experiments on the treatment of acute liver failure animal models by exosomes derived from mesenchymal stem cells were collected. Two evaluators screened the literature and extracted the data independently. The bias risk was evaluated by the SYRCLE tool. The extracted data were analyzed by Revmen 5.4.1 software and Stata 17.0 software.
    RESULTS: A total of 241 articles were retrieved and 9 animal experiments were included, with 219 animals: 110 animals in the model group and 109 animals in the exosome group. The results showed that the survival rate of animals in the exosome group improved significantly [RR=9.34, 95%CI(3.91, 22.29), P < 0.001], the levels of serum alanine transaminase [SMD=-5.31, 95%CI(-7.43, -3.19), P < 0.001] and aspartate aminotransferase [SMD=-4.47, 95%CI(-5.85, -3.10), P < 0.001] were reduced obviously. The expressions of interleukin-1β [SMD=-11.54, 95%CI(-18.12, -4.95), P=0.000 6], interleukin-6 [SMD=-5.75, 95%CI(-8.08, -3.41), P < 0.001] and tumor necrosis factor-α [SMD=-4.46, 95%CI(-6.83, -2.09), P=0.000 2], were suppressed obviously. 
    CONCLUSION: Exosomes derived from mesenchymal stem cells effectively inhibit the inflammatory response, ameliorate liver function of animals with acute liver failure, and improve their survival rate. The results of subgroup analysis showed that the shorter survival time of animals (≤ 24 hours), the lower dose of transplanted exosomes (< 1 mg/kg) and the source of exosomes (adipose-derived mesenchymal stem cells) may affect the efficacy of the exosomes derived from mesenchymal stem cells in the animal model of acute liver failure. This conclusion and its clinical transformation still need to be confirmed by randomized controlled studies with large sample sizes and high quality.
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    Visualization analysis of stem cell therapy for myocardial infarction based on Web of Science in recent ten years
    Sun Yukang, Song Lijuan, Wen Chunli, Ding Zhibin, Tian Hao, Ma Dong, Ma Cungen, Zhai Xiaoyan
    2024, 28 (7):  1143-1148.  doi: 10.12307/2024.138
    Abstract ( 407 )   PDF (3984KB) ( 192 )   Save
    BACKGROUND: Although traditional therapies, including drugs and surgery, cannot repair the damaged myocardial tissue, the mortality rate of myocardial infarction remains high. Stem cells provide the possibility to solve this problem due to their self-renewal and multi-directional differentiation potential. 
    OBJECTIVE: To analyze the research progress of stem cell therapy for myocardial infarction in recent ten years by bibliometric analysis. 
    METHODS: The related articles on stem cells and myocardial infarction published in SCI-E and SSCI from January 1, 2012 to December 1, 2022 in the Web of Science database were searched. EXCEL, CiteSpace and VOSviewer software were used to make statistical and visualization analyses of the data such as the number of publications, authors, institutions, journals, countries and keywords.  
    RESULTS AND CONCLUSION: A total of 3 210 core articles were published, and the total number increased year by year. hausenloy,derek j. is the author with the largest number of publications, China is the country with the largest number of publications, and the Fourth Military Medical University is the institution with the largest number of publications. The research hotspots in this field are changing from cell experiments and animal experiments to clinical trials. In the past ten years, research in this field has been highly popular and still has great development prospects. It is necessary to promote international and inter-agency exchange and learning, and further explore the role of stem cells in the treatment of myocardial infarction.
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