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    08 May 2024, Volume 28 Issue 13 Previous Issue    Next Issue
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    Biocompatibility and electrical output performance of composite piezoelectric film and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells
    Wang Zhen, Xu Shunen, Tang Geng, Luo Siwei, Teng Jianxiang, Xie Mengli, He Jialin, Ye Chuan
    2024, 28 (13):  1969-1975.  doi: 10.12307/2024.139
    Abstract ( 195 )   PDF (2294KB) ( 99 )   Save
    BACKGROUND: Adjustable piezoelectric effect can promote tissue regeneration and repair. Piezoelectric materials are widely used in weight-bearing tissue engineering.
    OBJECTIVE: To prepare a piezoelectric film material that can promote bone regeneration, and to explore its structural characterization, electrical output performance, biocompatibility, and effect of electrical output on osteogenic differentiation of rabbit bone marrow mesenchymal stem cells.
    METHODS: Using poly-3-hydroxybutyrateco/4-hydroxybutyrate (P34HB) as raw material, barium calcium stannate titanate powder (Ba0.94Ca0.06Sn0.08Ti0.92O3, BCST) was added according to mass ratios of 0%, 5%, 10%, 15%, and 20%. Dichloromethane was added to solve P34HB, and the thickness of 150-200 μm BCST/P34HB piezoelectric film was prepared by vacuum drying method. After polarization in the oil bath, the surface morphology, crystal phase composition, piezoelectric coefficient and open circuit voltage were tested. The effect of BCST/P34HB electrical output at 110 Hz and 0.25 N force on the proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells was tested.

    RESULTS AND CONCLUSION: (1) Scanning electron microscopy, X-ray diffraction, water contact angle, piezoelectric coefficient and electrical output performance tests showed that when the mass ratio of BCST increased to 20%, the BCST/P34HB piezoelectric film had good piezoelectric properties (d33=5.9 pC/N) and electrical output performance (180 mV), which was closer to the suitable range of 500 mV for electrical stimulation. (2) Live and dead staining showed that on the first day of co-culture, 15% group and 20% group showed less red fluorescence. On the 5th day of culture, the number of green fluorescence in each group was significantly higher than that on the first day, and the red fluorescence was not observed in the 10%, 15% and 20% groups, and only a small amount of red fluorescence was observed in the 0% and 5% groups. (3) On the 1st, 3rd and 5th days of co-culture with rabbit bone marrow mesenchymal stem cells, Almar blue staining exhibited that the number of cells in each group showed an increasing trend with the increase of time. On the 5th day of culture, the number of cells in the 20% group was significantly more than that in the 0% group (P < 0.05). (4) On day 10 of osteogenic induction, alkaline phosphatase staining results showed that the positive rate of the 20% group was significantly higher than that of the 0% group (P=0.000 1). On day 21, alizarin red staining and quantitative analysis of calcium nodules showed a similar trend to alkaline phosphatase staining. Compared with the 0% group, the 15% group and 20% group showed significant differences (P < 0.01, P < 0.000 1). (5) The results showed that 20% BCST/P34HB films had good piezoelectric properties, electrical output properties, biocompatibility and the ability of promoting osteogenic differentiation of bone marrow mesenchymal stem cells.

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    Chondrogenic differentiation of rabbit bone marrow mesenchymal stem cells induced by growth differentiation factor 5
    Li Feifei, Wang Buyu, Yang Zhihang, Dong Xiaoyu, Deng Jiang
    2024, 28 (13):  1976-1982.  doi: 10.12307/2024.126
    Abstract ( 180 )   PDF (1748KB) ( 14 )   Save
    BACKGROUND: Growth differentiation factor 5 is a member of the transforming growth factor superfamily and one of the earliest markers of joint development. Growth differentiation factor 5 has an important role in cartilage repair. 
    OBJECTIVE: To explore the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells. 
    METHODS: Rabbit bone marrow mesenchymal stem cells were isolated and cultured. CCK-8 assay was used to detect the effect of different mass concentrations of growth differentiation factor 5 on the proliferation activity of bone marrow mesenchymal stem cells. RT-PCR was utilized to detect the expression of genes related to chondrogenic differentiation of bone marrow mesenchymal stem cells induced by different mass concentrations of growth differentiation factor 5. To further investigate the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells, we added inhibitor XAV-939 and activator Laduviglusib of Wnt/β-catenin signaling pathways to induce cell culture for 14 days. RT-PCR and western blot assay were performed to detect the expression of cartilage-related genes and Wnt/β-catenin signaling pathway proteins.
    RESULTS AND CONCLUSION: (1) CCK-8 results showed no significant effect of growth differentiation factor 5 on the proliferation of bone marrow mesenchymal stem cells. (2) Growth differentiation factor 5 promoted the expression of cartilage-related genes type II collagen, aggrecan and Sox9, among which growth differentiation factor 5 induced a significant upregulation of cartilage-related genes in the 50 ng/mL group. (3) Addition of Laduviglusib, an activator of Wnt/β-catenin signaling pathway, upregulated Sox9, β-catenin and type II collagen expression (P < 0.05). Addition of XAV939, an inhibitor of Wnt/β-catenin signaling pathway, down-regulated Sox9, β-catenin and type II collagen expression (P < 0.05). (4) Taken together, growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells may be associated with the activation of the Wnt/β-catenin signaling pathway. 
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    Effects of micro-electric field on proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells
    Liu Zhong, Li Kewei, Wang Min, Liu Wenhui, Zhang Leilei, Guo Song, Qian Hui, Fu Qiang
    2024, 28 (13):  1983-1988.  doi: 10.12307/2024.161
    Abstract ( 205 )   PDF (1491KB) ( 17 )   Save
    BACKGROUND: Electrical stimulation is a physical method that can be used to induce various cellular activities such as cell proliferation, differentiation, and apoptosis. The induction of osteogenic differentiation of stem cells will be beneficial in the field of bone regeneration.
    OBJECTIVE: To observe whether micro-current field can promote the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells.
    METHODS: The fresh human umbilical cord tissue was cut to obtain umbilical cord mesenchymal stem cells, which were inoculated into a 6-well plate after cell culture and passage to the third generation. After 24 hours, the cells were cultured under a stimulation of 0, 50, and 100 mV/mm micro-electric field, at a frequency of 1 hour per day for 3 continuous days. The growth and morphological changes of human umbilical cord mesenchymal stem cells were observed by a microscope. The cell proliferation was detected by CCK-8 assay and EdU staining. Alizarin red staining was used to detect the osteogenic differentiation ability of cells. Western blot assay was used to determine the expression of ERK signal pathway proteins. 
    RESULTS AND CONCLUSION: (1) The optical density value and the number of proliferating cells in 50 and 100 mV/mm groups were significantly higher than those of the unstimulated group (P < 0.05). (2) Human umbilical cord mesenchymal stem cells could be induced to differentiate into osteocytes before and after micro-electric field stimulation, but the differentiation rate of 50 and 100 mV/mm groups was faster than that of unstimulated groups. (3) The protein expression of p-ERK1/2 in the 50 and 100 mV/mm groups was higher than that in the unstimulated group, and significant difference was detected between the 100 mV/mm group and the unstimulated group (P < 0.05). (4) Micro-electric field can promote the proliferation of human umbilical cord mesenchymal stem cells, and the mechanism may be achieved by promoting the phosphorylation of ERK.
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    Preparation and characterization of chitosan biguanide hydrochloride hydrogels loading mouse umbilical cord mesenchymal stem cells
    Deng Guodong, Yang Jia, Liu Yang
    2024, 28 (13):  1989-1995.  doi: 10.12307/2024.120
    Abstract ( 204 )   PDF (1612KB) ( 26 )   Save
    BACKGROUND: With the in-depth research of hydrogel materials, the applicable fields of hydrogel have been gradually broadened, and carrying stem cells for disease treatment has become a new direction of research, but how to construct a hydrogel suitable for stem cell growth is the key problem that needs to be solved at present.
    OBJECTIVE: To investigate the physicochemical properties of chitosan-chitosan biguanide hydrochloride-collagen composite hydrogels and to evaluate their ability to load mouse umbilical cord mesenchymal stem cells. 
    METHODS: The hydrogels were prepared by physically cross-linking chitosan, chitosan biguanide hydrochloride and collagen with the cross-linking agents β-glycerophosphate sodium and sodium bicarbonate, and the suitable hydrogels were screened according to the gel formation time and gel formation effect (noted as Gel-1, Gel-2 and Gel-3 in this way). Morphology, porosity, swelling properties, and degradability of the three groups of hydrogels were observed by scanning electron microscopy. Hemolysis experiments were performed to examine the hemolysis of the three groups of hydrogels. The mouse umbilical cord mesenchymal stem cells were co-cultured with the hydrogel with the best comprehensive performance of characterization. The cytotoxicity, cell survival and adhesion effect of the composite hydrogel were determined to evaluate the performance of this hydrogel loaded with umbilical cord mesenchymal stem cells. 
    RESULTS AND CONCLUSION: (1) Scanning electron microscopy characterization results showed that all three groups had porous mesh structures inside, and the internal structure of Gel-2 and Gel-3 with the addition of chitosan biguanide hydrochloride was more porous and three-dimensional. (2) The hydrogel porosity of the Gel-3 group was higher than the remaining two groups, with high porosity and uniform pore size distribution. (3) The swelling performance of all three groups of hydrogels was above 100%, and the swelling performance of hydrogels with chitosan biguanide hydrochloride component was better. (4) The degradation rate of the three groups of hydrogels could be degraded by more than 90% in a time scale of 15 days, with good degradation performance. (5) The results of the hemolytic properties showed that the absorbance values measured by each group of hydrogels carrying chicken erythrocytes were basically the same as those of saline  carrying chicken erythrocytes, and no hemolysis occurred. (6) The toxicity experiment and living and dead cell staining showed that the survival rate of umbilical cord mesenchymal stem cells in each group of hydrogels was above 100%, indicating that there was no obvious cytotoxicity. Umbilical cord mesenchymal stem cells could survive under the hydrogel package and the hydrogels had a positive effect on the survival rate of umbilical cord mesenchymal stem cells. (7) The cells in the umbilical cord mesenchymal stem cell adhesion assay can survive under the hydrogel package and can adhere to the surface of the hydrogel with normal morphology.
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    Wnt signaling pathway is involved in differentiation of embryonic stem cells into thymic epithelial progenitor cells together with autoimmune regulators
    Wen Tinghao, Li Yuandi, He Keke, Song Wenqian, Wang Xianbin, Gao Jie, Su Min, Hu Rong
    2024, 28 (13):  1996-2001.  doi: 10.12307/2024.132
    Abstract ( 153 )   PDF (1120KB) ( 31 )   Save
    BACKGROUND: Autoimmune regulator gene (Aire) and Wnt signaling pathway play an important role in the maintenance and differentiation of mouse embryonic stem cell pluripotency. However, whether the Wnt signal and Aire are involved in the differentiation of embryonic stem cells to thymic epithelial progenitor cells remains poorly understood. 
    OBJECTIVE: To investigate the relationship of the Wnt signaling pathway and Aire with the differentiation of embryonic stem cells. 
    METHODS: A two-step differentiation method was used to induce mouse embryonic stem cells to differentiate into endoderm and then into thymic epithelial progenitor cells. Mouse embryonic stem cells were infected with Aire shRNA lentivirus, and monoclonal stable strains were screened by puromycin. Mouse embryonic stem cells were collected on days 0, 3 and 10 of the directed induction of differentiation after the induced differentiation by the two-step differentiation method. Cellular immunofluorescence, flow cytometry, western blot assay, and real-time qPCR were used to detect the expression changes of related genes and proteins.
    RESULTS AND CONCLUSION: (1) Immunofluorescence staining showed positive expression of SSEA1 and OCT4 on day 0 of targeted induction of differentiation. (2) Immunofluorescence staining showed double-positive expression of SOX17 and FOXA2 on day 3 of targeted induction of differentiation. (3) Flow cytometry results showed positive expression of EPCAM1, K5 and K8 on day 10 of targeted induction of differentiation. (4) Compared with undifferentiated mouse embryonic stem cells, the expressions of Wnt7a, β-catenin, and Gsk-3β proteins were elevated, and the expression level of Aire protein was decreased in induced differentiated thymic epithelial progenitor cells. (5) Compared with undifferentiated mouse embryonic stem cells, the expressions of Wnt7a, β-catenin, Gsk-3β and Aire mRNA were elevated in thymic epithelial progenitor cells. (6) Compared with normal cultured mouse embryonic stem cells and their ultimately differentiated thymic epithelial progenitor cells, the expression levels of Wnt7a, β-catenin and Gsk-3β proteins were reduced in mouse embryonic stem cells with knockdown of Aire genes and their final differentiated thymic epithelial progenitor cells. In conclusion, the Wnt signaling pathway and Aire are jointly involved in the process of targeted induction of differentiation of mouse embryonic stem cells into mouse thymic epithelial progenitor cells. 
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    Radioprotective effect of 1,2-propanediol combined with hepatocyte growth factor-modified dental pulp stem cell exosomes on human skin cells
    Liu Yun, Jin Jiayan, Liu Yubin, Li Qiang, Ren Boyuan, Wu Zuze, Zhou Gangqiao, Jin Jide
    2024, 28 (13):  2002-2008.  doi: 10.12307/2024.169
    Abstract ( 228 )   PDF (1250KB) ( 52 )   Save
    BACKGROUND: Skin damage caused by radiation therapy and nuclear accidents is still a serious medical problem. It is difficult to achieve effective treatment results with single prevention and treatment methods. It is an important research direction to find new comprehensive treatment methods. 
    OBJECTIVE: To observe the protective effect and the underlying mechanism of 1,2-propanediol combined with hepatocyte growth factor-modified exosomes derived from dental pulp stem cells on human epidermal radiation damage cell models.
    METHODS: (1) After infection of human dental pulp stem cells using recombinant adenovirus of human hepatocyte growth factor gene, exosomes, i.e., Ad. HGF DPSC-Exo, were isolated with ultracentrifugation. (2) HaCat cells were irradiated with X-ray. The cells were treated with 1,2-propanediol before irradiation and Ad.HGF DPSC-Exo after irradiation. Cell proliferative activity was determined by CCK-8 assay. Cell apoptosis was detected by flow cytometry. Cell migration was detected by cell scratch assay. The expression levels of P21 and P53 were detected by PCR.
    RESULTS AND CONCLUSION: 1,2-Propanediol, Ad.HGF.DPSC-Exo, Ad.HGF.DPSC-Exo + 1,2-propanediol could significantly improve the growth inhibition of HaCaT cells, reduce cell apoptosis, elevate cell proliferation and migration, and exhibit a good radiation protection effect. Moreover, the combined effect of Ad.HGF.DPSC-Exo + 1,2-propanediol was better. Furthermore, Ad.HGF.DPSC-Exo + 1,2-propanediol alleviated the cellular G2/M phase block and decreased the expression of cell cycle genes P53 and P21. In conclusion, 1,2-propanediol pretreatment combined with Ad.HGF.DPSC-Exo had significant protective effects on radiation-induced HaCaT cell injury and it provided novel ideas and potential methods for the prevention and treatment of radiation-induced skin damage. 
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    Analysis of risk factors for short-term death after allogeneic hematopoietic stem cell transplantation
    Gao Siyu, Yao Lihong, Bian Zhilei, Zhang Suping, Li Li, Fan Jinpeng, Qin Jing, Peng Yingnan, Wan Dingming
    2024, 28 (13):  2009-2016.  doi: 10.12307/2024.173
    Abstract ( 240 )   PDF (1281KB) ( 67 )   Save
    BACKGROUND: Allogeneic hematopoietic stem cell transplantation is an effective and even the only way to cure various hematological diseases, but the short-term mortality rate is relatively high after transplantation.
    OBJECTIVE: To investigate the risk factors affecting the overall survival of patients with hematological diseases in the short term (within 100 days) after allogeneic hematopoietic stem cell transplantation, so as to reduce mortality and effectively prevent related risks in the short term (within 100 days) after allogeneic hematopoietic stem cell transplantation.
    METHODS: Clinical data of 585 patients with hematological diseases who underwent allogeneic hematopoietic stem cell transplantation at the Hematopoietic Stem Cell Transplantation Center of First Affiliated Hospital of Zhengzhou University from January 1, 2018 to June 30, 2021 were retrospectively analyzed. The risk factors that affected overall survival within 100 days after allogeneic hematopoietic stem cell transplantation were explored.
    RESULTS AND CONCLUSION: A total of 585 patients with hematologic diseases underwent allogeneic hematopoietic stem cell transplantation. 92 patients died within 100 days after transplantation, with a mortality rate of 15.7% (92/585). The median age of death cases was 26.5 years old (1-56 years), and the median survival time of death cases was 48 days (0-97 days). Univariate analysis exhibited that age ≥14 years old, acute graft-versus-host disease, grade IV acute graft-versus-host disease, bacterial bloodstream infection, as well as carbapenem-resistant organism bloodstream infection, were risk factors for overall survival within 100 days after allogeneic hematopoietic stem cell transplantation (P < 0.05). Multivariate regression analysis showed that age ≥14 years old, grades III-IV acute graft-versus-host disease, bacterial bloodstream infection, and carbapenem-resistant organism bloodstream infections were independent risk factors for overall survival (within 100 days) in patients after allogeneic hematopoietic stem cell transplantation. Hazard ratios were 1.77(95%CI 1.047-2.991), 7.926(95%CI 3.763-16.695), 2.039(95%CI 1.117-3.722), and 3.389(95%CI 1.563-7.347), respectively. In conclusion, all-cause mortality rate after allogeneic hematopoietic stem cell transplantation is relatively high in the short term. A timely diagnosis and effective treatment of bacterial bloodstream infection and acute graft-versus-host disease are essential to improving allogeneic hematopoietic stem cell transplantation outcomes.
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    Effect of miR-26b on proliferation, migration and osteogenic differentiation of stem cells from human exfoliated deciduous teeth and human umbilical cord mesenchymal stem cells
    Yuan Yuanyuan, Pan Lu, Zhou Shaolan, Liang Yan, Xu Jianwei, Xu Wen
    2024, 28 (13):  2017-2023.  doi: 10.12307/2024.166
    Abstract ( 193 )   PDF (2333KB) ( 19 )   Save
    BACKGROUND: microRNA-26b (miR-26b) plays an important regulatory role in a variety of stem cell functions, but its effects on the biological properties of stem cells from human exfoliated deciduous teeth and human umbilical cord mesenchymal stem cells are unknown.
    OBJECTIVE: To investigate the effects of miR-26b on the proliferation, migration and osteogenic differentiation of stem cells from human exfoliated deciduous teeth and human umbilical cord mesenchymal stem cells. 
    METHODS: Stem cells from human exfoliated deciduous teeth and human umbilical cord mesenchymal stem cells were cultured and identified. miR-26 mimics (experimental group) and miRNAs mimics control (control group) were used to transfect above mentioned two kinds of cells and construct overexpressed models for subsequent experiments. CCK-8 assay was applied to detect the proliferation ability of overexpressed miR-26b cells. Transwell and scratch assay were employed to analyze the migration ability of overexpressed miR-26b cells. RT-qPCR was utilized to examine the expression of osteogenic markers after osteogenic induction of overexpressed miR-26b cells.
    RESULTS AND CONCLUSION: (1) Transfection of miR-26b mimics increased miR-26b expression in the two kinds of cells and promoted the proliferation of stem cells from human exfoliated deciduous teeth, with no significant effect on the amplification of human umbilical cord mesenchymal stem cells. (2) Compared with the control group, the migration ability was enhanced after two types of cells overexpressing miR-26b. (3) miR-26b expression decreased during osteogenic differentiation of the two kinds of cells. (4) Compared with the control group, the levels of osteogenesis-related genes osteocalcin, osteopontin, alkaline phosphatase, and human type I collagen mRNA were downregulated after overexpression of miR-26b in the two kinds of cells. The results showed that overexpression of miR-26b promoted the proliferation and migration of stem cells from human exfoliated deciduous teeth and inhibited their osteogenic differentiation; it promoted the migration of human umbilical cord mesenchymal stem cells and inhibited their osteogenic differentiation, but had no significant effects on their proliferation.
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    Effects of different culture media on quality and developmental potential of mouse oocytes after in vitro maturation
    Tian Yin, Zhao Yanhua, Huang Guoning, Li Jingyu
    2024, 28 (13):  2024-2029.  doi: 10.12307/2024.124
    Abstract ( 236 )   PDF (1715KB) ( 27 )   Save
    BACKGROUND: In recent years, the demand for in vitro maturation of immature oocytes has increased. Oocyte maturation is affected by many factors, among which the selection of medium is particularly important, and there is currently no unified plan. 
    OBJECTIVE: To compare the in vitro maturation of germinal vesicle stage oocytes with different maturation media and to investigate its effects on oocyte quality and developmental potential. 
    METHODS: Germinal vesicle oocytes were matured in G-1TM PLUS medium, CZB medium and M16 medium, and mature oocytes in vivo were used as control group to compare in vitro fertilization and early embryo development among various groups. The immunofluorescence method was used to evaluate mitochondrial function in mature oocytes of each group. Calcium oscillation was detected by confocal microscopy real-time imaging system.
    RESULTS AND CONCLUSION: (1) There was no significant difference in the first polar body ejection rate among the three groups (P > 0.05). (2) The rate of in vitro fertilization was higher in the G-1TM PLUS group (52.86±11.24)% than that in the M16 group (37.76±6.70)% and the CZB group (30.62±5.51)%. The blastocyst rate was lower in the CZB group (36.23±6.63)% than that in the control group (78.16±4.17)%, G-1TM PLUS group (55.75±7.63)% and M16 group (53.36±6.33)%. (3) Compared with the control group, the length-to-width ratio of the spindle in the CZB group increased (P < 0.005). (4) The mitochondrial function of the CZB group was worse than that of the control group, G-1TM PLUS group and M16 group, and abnormal mitochondrial agglutination occurred in the CZB group. (5) The frequency of calcium oscillations in the CZB and M16 groups was significantly higher than that in the G1 and control groups. In conclusion, during in vitro maturation of mouse oocytes, in vitro maturation rate was not significantly different among G-1TM PLUS, CZB and M16 media, but the G-1TM PLUS medium had a higher rate of fertilization and blastocyst formation.
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    Silencing of activating transcription factor 4 inhibits neuronal necroptosis after intracerebral hemorrhage in vitro
    Feng Dengfeng, Wei Min, Zhang Hengzhu
    2024, 28 (13):  2030-2035.  doi: 10.12307/2024.147
    Abstract ( 210 )   PDF (2189KB) ( 28 )   Save
    BACKGROUND: Neuronal necroptosis induced by intracerebral hemorrhage is an important cause of secondary brain injury. Activating transcription factor 4 (ATF4) is a member of the transcription activator family, which plays an important role in secondary brain injury after intracerebral hemorrhage. However, the mechanism of ATF4 in neuronal necroptosis after intracerebral hemorrhage remains unclear. 
    OBJECTIVE: To explore the effect of ATF4 silencing (ATF4 small interfering RNA, ATF4 siRNA) on neuronal necroptosis after intracerebral hemorrhage.
    METHODS: The HT-22 mouse hippocampal neuron cell line and the BV-2 mouse microglial cell line were co-cultured, and hemin was used to mimic an in vitro model of intracerebral hemorrhage. A gradient concentration of hemin was used to treat cells and was set in the interval of 0-100 μmol/L, and the cell viability was evaluated by MTT assay after 24 hours of administration of hemin. The cells were divided into four groups: the blank control group without any intervention; the control group was treated with hemin (50 μmol/L), and the other two groups were treated with negative control small interfering RNA (NC siRNA) and ATF4 small interfering RNA (ATF4 siRNA) 48 hours before administration of hemin. After the cells were treated with hemin (50 μmol/L) for 24 hours, PI/Hoechst staining was used to detect neuronal necroptosis. Western blot assay was used to detect the protein expression of ATF4, receptor-interacting protein 3 (RIP3), and mixed lineage kinase domain-like protein (MLKL), and double immunofluorescent staining was located in neurons to observe the level of neuronal necroptosis and the regulatory effect of ATF4 on it.
    RESULTS AND CONCLUSION: (1) 50 μmol/L of hemin could induce neuronal necroptosis to a greater extent. (2) The number of PI+/Hoechst+ cells in the control group and NC siRNA group was higher than that in the blank control group (P < 0.000 1). The number of PI+/Hoechst+ cells in the ATF4 siRNA group was lower than that in the control group (P < 0.000 1). (3) Compared with the control group, the ATF4 siRNA group not only inhibited the expression of ATF4 protein (P < 0.001), but also inhibited the expression of RIP3 and MLKL protein (P < 0.001). (4) Through double immunofluorescent staining, compared with the control group, the protein expression of RIP3 and MLKL was significantly reduced in the ATF4 siRNA group (P < 0.000 1). (5) The results show that the silencing of the ATF4 gene can directly or indirectly inhibit the expression of genes related to neuronal necroptosis after intracerebral hemorrhage, and play a vital role in alleviating secondary brain injury.
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    Overexpression of miR-378a promotes macrophage M2 polarization and inhibits M1 polarization
    Yang Quan, He Huiyu, Wang Sifan, Lyu Shangyi, Zhou Qiqi, Han Xiangzhen
    2024, 28 (13):  2036-2041.  doi: 10.12307/2024.141
    Abstract ( 175 )   PDF (1207KB) ( 81 )   Save
    BACKGROUND: M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing. Therefore, how to regulate M2 polarization of macrophages has been a hot research topic in recent years, and some miRNAs have been found to have this function.
    OBJECTIVE: To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line.
    METHODS: The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ. Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages. By transfection with lentivirus as the vector of overexpression of miR-378a, the stable expression of miR-378a cell lines was screened. Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ. Macrophage M2 polarization was induced by interleukin-4. The levels of M1/M2 polarization-related cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay. qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines. 
    RESULTS AND CONCLUSION: (1) The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization. (2) Compared with the non-transfected group, the expressions of proinflammatory cytokine-induced nitric oxide synthase, tumor necrosis factor-α, interleukin-6 and interleukin-1β in macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group (P < 0.05); the levels of inducible nitric oxide synthase, tumor necrosis factor-α and interleukin-6 in cell supernatant were also significantly decreased (P < 0.05). (3) Compared with the non-transfected group, the expressions of CD206, interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased (P < 0.05); the levels of CD206 and interleukin-10 in cell supernatant were also significantly increased (P < 0.05) in the miR-378a transfection group. (4) It is indicated that overexpression of miR-378a promotes the M2 polarization of macrophages and inhibits the M1 polarization of macrophages. 
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    Efficacy of umbilical cord mesenchymal stem cells replacing donor bone marrow cells in haploidentical hematopoietic stem cell transplantation
    Zhang Wenhui, Pei Xiaohang, Kong Dai, Liu Zhongwen
    2024, 28 (13):  2042-2046.  doi: 10.12307/2024.170
    Abstract ( 190 )   PDF (803KB) ( 20 )   Save
    BACKGROUND: HLA haploid allogeneic hematopoietic stem cell transplantation provides a chance of survival for patients with high-risk hematologic malignancies. In recent years, the research on the transplantation mode and graft selection of haploidentical transplantation is still ongoing. At present, the mixed transplantation model of non-extracorporeal T-cell removal bone marrow and peripheral blood stem cells established by the Hematology Research Center of Peking University is gradually becoming popular in China, but this model requires the collection of donor bone marrow fluid, which increases the pain and risk of the donor.
    OBJECTIVE: To explore the curative effect of infusion of umbilical cord mesenchymal stem cells replacing donor bone marrow cells in haploidentical peripheral blood hematopoietic stem cell transplantation for malignant hematological diseases.
    METHODS: Fifty hematological malignancies patients who underwent haploidentical hematopoietic stem cell transplantation from January 2019 to May 2022 were selected and randomly assigned to two study groups at a ratio of 2:3. Among them, 19 patients received umbilical cord mesenchymal stem cell combined with peripheral blood stem cell transplantation, and 31 patients were treated with bone marrow cells combined with peripheral blood stem cells. The study was approved by the Ethics Committee of Henan Provincial People’s Hospital. The recipients of umbilical cord mesenchymal stem cells were first transfused with third-party umbilical cord mesenchymal stem cells (1×106/kg) on the day of transplantation, followed by peripheral blood hematopoietic stem cells 6 hours later. In the bone marrow group, donor bone marrow cells were transfused +1 day after transplantation and peripheral blood stem cells were transfused +2 days after transplantation. After transplantation, rabbit anti-human thymocyte immunoglobulin, cyclosporine A, mycophenolate mofetil, and a short-course methotrexate were used for graft-versus-host disease prophylaxis for all recipients.
    RESULTS AND CONCLUSION: No adverse events occurred during the reinfusion of umbilical cord mesenchymal stem cells. There were no significant differences between the mesenchymal stem cell group and the bone marrow group in the engraftment rate [100% (19/19) vs. 96.8%(30/31), P > 0.05], median duration for neutrophil engraftment (14 days vs.15 days, P > 0.05) and median duration for platelet engraftment (20 days vs.19 days, P > 0.05). The incidence of grade II-IV acute graft-versus-host disease in the mesenchymal stem cell group was significantly lower than in the bone marrow group [21.1% (4/19) vs. 58.1% (18/31), P = 0.01]. There were no significant differences between the two groups in the incidence of chronic graft-versus-host disease [21.1% (4/19) vs. 25.8% (8/31), P > 0.05], the relapse rates [15.8% (3/19) vs. 16.1% (5/31), P > 0.05] and the incidence of early cytomegalovirus viremia [42.1% (8/19) vs. 35.5% (11/31), P > 0.05], and the 2-year overall survival rate [68.4% (10/19) vs. 70.9% (16/31), P > 0.05]. It is indicated that umbilical cord mesenchymal stem cells replace donor bone marrow cells in haploidentical peripheral blood stem cell transplantation for malignant hematological diseases, which reduced the incidence of acute graft-versus-host disease after transplantation, did not increase the incidence of chronic graft-versus-host disease, recurrence rate and early cytomegalovirus viremia, and reduced the pain and risk of donor pulp extraction.
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    Placenta-derived mesenchymal stem cells in promoting acute skin wound healing in rats
    Dong Hongfei, Huang Xi, Li Xianhui, Zhang Yanbiao, Wang Xuyang, Wang Bing, Sun Hongyu
    2024, 28 (13):  2047-2053.  doi: 10.12307/2024.136
    Abstract ( 178 )   PDF (3758KB) ( 21 )   Save
    BACKGROUND: Currently, a variety of mesenchymal stem cells have been confirmed to have the effect of promoting wound repair, but there is still a lack of relevant research on whether placenta-derived mesenchymal stem cells can promote acute skin wound healing.
    OBJECTIVE: To investigate the effect of placenta-derived mesenchymal stem cell transplantation on the healing of acute skin wound in rats.
    METHODS: Twenty SD rats were divided into PBS group and stem cell group by the random number table method, with 10 rats in each group. All rats were selected to establish a full-thickness skin defect model. In the PBS group and stem cell group, PBS buffer and placenta-derived mesenchymal stem cells were immediately injected on the wound surface and wound margin immediately and on day 8 after modeling. The wound healing was observed immediately and on days 2, 4, 6, 8, 10, 12, and 14 after modeling. The skin tissue of the wound surface was taken on day 14 and treated with hematoxylin-eosin staining, Masson staining, immunohistochemical staining and immunofluorescence staining. 
    RESULTS AND CONCLUSION: (1) The wound surface of the rats in each group decreased with the prolongation of treatment time. The wound healing rate and wound epithelization rate of the stem cell group at 14 days were higher than those of the PBS group (P < 0.01), and the wound contracture rate was lower than that of the PBS group (P < 0.01). (2) The results of hematoxylin-eosin staining showed that the skin wound healing of the stem cell group was better than that of the PBS group; the degree of wound epithelization was higher, and the morphology of collagen fibers was close to that of normal skin. (3) Masson staining results showed that compared with the PBS group, collagen fibers in the skin wound tissue of the stem cell group were significantly increased and thicker, and the content of collagen fibers in the new tissue was significantly higher than that of the PBS group (P < 0.01). (4) Immunohistochemical staining showed that the number of new capillaries in the stem cell group was higher than that in the PBS group (P < 0.01), while the expressions of tumor necrosis factor-α and interleukin-6 were lower than those in the PBS group (P < 0.01). (5) Immunofluorescence staining showed that the number of M2 macrophages in the new wounds of the stem cell group was higher than that of the PBS group (P < 0.01), while the number of M1 macrophages was less than that in the PBS group (P < 0.01). These findings indicate that placenta-derived mesenchymal stem cells can accelerate skin wound healing, promote wound epithelization, and reduce wound contracture, which may be related to the promotion of capillary angiogenesis, regulation of collagen fiber production, inhibition of inflammation, and regulation of macrophage polarization to M2 type.
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    Hypoxic postconditioning protects myocardium by regulating autophagy in aging cardiomyocytes through piRNA-005854
    Chi Hongyang, Yang Huixia, Hao Yinju, Yang Anning, Bai Zhigang, Jiao Yun, Xiong Jiantuan, Ma Shengchao, Jiang Yideng
    2024, 28 (13):  2054-2060.  doi: 10.12307/2024.133
    Abstract ( 174 )   PDF (1285KB) ( 20 )   Save
    BACKGROUND: Ischemic postconditioning is one of the effective ways to reduce ischemia-reperfusion injury and has been more and more widely used in clinical practice in recent years, but its specific molecular mechanism has yet to be studied.  
    OBJECTIVE: To investigate the role and mechanism of piRNA-005854 in the aging cardiomyocytes caused by hypoxic postconditioning.
    METHODS: In vitro, cardiomyocytes were administered 8 mg/mL D-galactose for 9 days to induce their aging. β-Galactosidase staining was used to observe the aging of cardiomyocytes. Senescent cells were treated with hypoxia/reoxygenation and hypoxic postconditioning. ELISA was utilized to detect changes in myocardial injury markers creatine kinase isoenzyme MB and lactate dehydrogenase levels. Western blot assay was applied to detect the expression changes of autophagy-related proteins LC3II, p62, ULK1 and phosphorylated ULK1 in aging cardiomyocytes. qRT-PCR was employed to determine the expression level of piRNA-005854. piRNA-005854 inhibitor and piRNA-005854 mimics were transferred into aging cardiomyocytes and followed with hypoxic postconditioning. Western blot assay was used to examine the expression of LC3II, p62, ULK1 and phosphorylated ULK1.  
    RESULTS AND CONCLUSION: (1) D-galactose induced obvious senescence of cardiomyocytes 9 days later. (2) Compared with the normoxia group, creatine kinase isoenzyme MB and lactate dehydrogenase levels increased in the hypoxia/reoxygenation group (P < 0.01); LC3 II/I expression was increased; p62 expression was decreased; ULK1 phosphorylation level was increased, and piRNA-005854 expression was increased (P < 0.01). (3) Compared with the hypoxia/reoxygenation group, creatine kinase isoenzyme MB and lactate dehydrogenase levels significantly reduced in the hypoxic postconditioning group (P < 0.01); LC3 II/I expression significantly decreased (P < 0.05); p62 expression increased (P < 0.01); ULK1 phosphorylation level decreased (P < 0.05), and piRNA-005854 expression decreased (P < 0.01). (4) After transfection of piRNA-005854 inhibitor, LC3II/I expression was decreased (P < 0.01); the expression of p62 was increased significantly (P < 0.05); the phosphorylation level of ULK1 was decreased significantly (P < 0.01). After transfection of piRNA-005854 mimics, LC3II/I expression was increased significantly; the expression of p62 was decreased, and the phosphorylation level of ULK1 was increased significantly (P < 0.01). (5) The results show that piRNA-005854-mediated reduction of ULK1-dependent autophagy level is a possible mechanism that hypoxic postconditioning exerts its protective effect on aging cardiomyocytes.
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    Ginsenoside Rg1 inhibits H2O2-induced apoptosis and autophagy in human periodontal ligament cells
    Yang Wanrong, Chu Yi, Xu Yao, Li Sihui, Guo Ling
    2024, 28 (13):  2061-2067.  doi: 10.12307/2024.121
    Abstract ( 198 )   PDF (1304KB) ( 39 )   Save
    BACKGROUND: Our previous studies have confirmed that H2O2 and ginsenoside Rg1 can cause changes in reactive oxygen species levels in human periodontal ligament cells, but the correlation of reactive oxygen species with apoptosis and autophagy remains unclear.
    OBJECTIVE: To explore the effect and mechanism of ginsenoside Rg1 on H2O2-induced apoptosis and autophagy of human periodontal ligament cells. 
    METHODS: Human periodontal ligament cells were divided into control group, H2O2 group and H2O2+Rg1 group. Ginsenoside Rg1 (50 μmol/L) was pre-incubated for 24 hours and H2O2 was treated for 2 hours (500 μmol/L). CCK-8 assay was used to detect the proliferation ability of the cells. A fluorescent probe DCFH-DA was used to detect the reactive oxygen species level of the cells. qRT-PCR and western blot assay were used to detect heme oxygenase 1, apoptosis-related factor Caspase-3, Bax, anti-apoptotic factor Bcl-2, autophagy Beclin-1, P62, LC3, pathway-related factors phosphatidylinositol-3-kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR) mRNA and protein expression.
    RESULTS AND CONCLUSION: (1) Compared with the control group, the proliferation activity of human periodontal ligament cells in the H2O2 group decreased. Compared with the H2O2 group, the proliferation activity of human periodontal ligament cells increased in the H2O2+Rg1 group. (2) Compared with the control group, the expression of reactive oxygen species and heme oxygenase-1 increased in the H2O2 group. Compared with the H2O2 group, the expression of reactive oxygen species and heme oxygenase-1 decreased in the H2O2+Rg1 group. (3) Compared with the control group, the expression of Caspase-3, Bax, Beclin-1 and LC3 increased, while the expression of Bcl-2 and P62 decreased in human periodontal ligament cells of the H2O2 group. Compared with the H2O2 group, the expressions of Caspase-3, Bax, Beclin-1 and LC3 decreased, and the expressions of Bcl-2 and P62 increased in the H2O2+Rg1 group. (4) Compared with the control group, the expressions of PI3K, AKT and mTOR decreased in human periodontal ligament cells of the H2O2 group. Compared with the H2O2 group, the expressions of PI3K, AKT and mTOR increased in human periodontal ligament cells of the H2O2+Rg1 group. (5) These results suggest that ginsenoside Rg1 can inhibit H2O2-induced apoptosis and autophagy in human periodontal ligament cells by reducing the content of reactive oxygen species and down-regulating the related factor expression of the PI3K/AKT/mTOR pathway.
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    Effects of aerobic exercise on adult hippocampal neurogenesis in Alzheimer’s disease mice
    Zhang Yeting, Li Chuikun, Wei Cuilan, Fu Yan, Zhang Feifei
    2024, 28 (13):  2068-2075.  doi: 10.12307/2024.131
    Abstract ( 196 )   PDF (2724KB) ( 28 )   Save
    BACKGROUND: Exercise improves Alzheimer’s disease, dementia, and age-related cognitive abilities. A potential mediator between exercise and these health benefits may be adult hippocampal neurogenesis. Therefore, it is of great significance to explore whether and how exercise affects the adult hippocampal neurogenesis process in Alzheimer’s disease mice. 
    OBJECTIVE: To observe the effect of aerobic exercise on adult hippocampal neurogenesis of Alzheimer’s disease mice, and to explore whether aerobic exercise can promote their adult hippocampal neurogenesis.
    METHODS: Three-month-old wild-type (C57BL/6Jnju) and APP/PS1 double transgenic Alzheimer’s disease mice were randomly divided into four groups: wild control group, wild exercise group, Alzheimer’s disease control group and Alzheimer’s disease exercise group, with 20 mice in each group. The control group did not do exercise, and the exercise group did aerobic exercise for 5 months. After exercise intervention, real-time PCR, immunofluorescence and western blot assay were used to detect the expression levels of DCX, Ki67, βIII-tubulin and NeuN in the hippocampal tissue of mice in each group. 
    RESULTS AND CONCLUSION: The expressions of DCX, βIII-tubulin and NeuN in the hippocampal dentate gyrus in the Alzheimer’s disease control group were significantly lower than those in the wild control group (P < 0.05). The expressions of DCX, Ki67, βIII-tubulin and NeuN were significantly higher in the hippocampal dentate gyrus in the Alzheimer’s disease exercise group than those in the Alzheimer’s disease control group (P < 0.05). It is indicated that long-term aerobic exercise intervention can strengthen the proliferation, migration and differentiation of neurons during adult hippocampal neurogenesis and significantly increase the number of neuronal precursor cells and new neurons in Alzheimer’s disease mice. 
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    Solute carrier family 2 member 12 intervenes in uric acid-induced renal tubular cell injury
    He Yi, Li Xiaolin, He Jinke, Jiang Xiangju, Liang Meiting, Chen Wujin, Cui Yuena, Sun Yuping
    2024, 28 (13):  2076-2081.  doi: 10.12307/2024.117
    Abstract ( 172 )   PDF (1381KB) ( 20 )   Save
    BACKGROUND: In recent years, the incidence of hyperuricemia caused by purine metabolism disorders has been increasing, which can induce inflammatory responses and lead to renal injury.  
    OBJECTIVE: To explore the role and mechanism of solute carrier family 2 member 12 (SLC2A12) in hyperuricemia-related renal injury. 
    METHODS: Renal tubular cells (HK2 cells) were divided into five groups: HK2 group, HK2+uric acid group, HK2+uric acid+NC group, HK2+uric acid+siSLC2A12 group, and HK2+uric acid+siSLC2A12+MK-2206 group. HK2 cells were treated with uric acid and transfected with siRNA SLC2A12, followed by MK-2206 treatment to inhibit AKT expression. Cell proliferation was detected by CCK-8 assay. Apoptosis was detected by TUNEL assay. qRT-PCR and western blot assay were used to detect fibrogenic factors as well as activation of the AKT/FOXO3a pathway. The concentrations of inflammatory cytokines were measured by enzyme-linked immunosorbent assay. 
    RESULTS AND CONCLUSION: (1) Uric acid treatment inhibited cell proliferation and promoted cell apoptosis in the HK2+uric acid group compared with the HK2 group. The proliferative ability of cells in the HK2+uric acid+siSLC2A12 group was further decreased and apoptotic cells were further increased compared with the HK2 group. Compared with the HK2+uric acid+siSLC2A12 group, the HK2+uric acid+siSLC2A12+MK-2206 group showed an increase in cell proliferation and a decrease in apoptotic cells. (2) Compared with the HK2 group, the connective tissue growth factor (CTGF), α-smooth muscle actin (α-SMA) and transforming growth factor beta (TGF-β) expressions increased in the HK2+uric acid group; CTGF, α-SMA and TGF-β expression further increased in the HK2+uric acid+siSLC2A12 group. Compared with the HK2+uric acid+siSLC2A12 group, the CTGF, α-SMA and TGF-β expressions decreased. (3) Compared with the HK2 group, the expression of p-AKT, FOXO3a, and p-FOXO3a elevated in the HK2+uric acid group; the expression of p-AKT further increased, while the expression of FOXO3a and p-FOXO3a decreased in the HK2+uric acid+siSLC2A12 group. Compared with the HK2+uric acid+siSLC2A12 group, p-AKT expression decreased; FOXO3a and p-FOXO3a expression increased in the HK2+uric acid+siSLC2A12+MK-2206 group. (4) Compared with the HK2 group, interleukin-6, interleukin-1 β, and tumor necrosis factor α levels increased in the HK2+uric acid group; interleukin-6, interleukin-1 β, and tumor necrosis factor α levels further increased in the HK2+uric acid+siSLC2A12 group. Compared with the HK2+uric acid+siSLC2A12 group, interleukin-6, interleukin-1 β, and tumor necrosis factor α levels diminished in the HK2+uric acid+siSLC2A12+MK-2206 group. (5) These findings indicate that SLC2A12 may protect against hyperuricemia-induced renal injury by counteracting uric acid-induced tubular fibrosis and inflammation through activation of the FOXO3a pathway.
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    Screening and verification of genes related to immune infiltration between myelodysplastic syndrome and acute myeloid leukemia
    Deng Fahua, Hu Huali, Wang Siqi, Xu Jianxia, Lu Tingting, Huang Hai, Wei Sixi
    2024, 28 (13):  2082-2089.  doi: 10.12307/2024.154
    Abstract ( 194 )   PDF (2965KB) ( 22 )   Save
    BACKGROUND: Myelodysplastic syndrome has worse hazards of acute myeloid leukemia transformation, and some studies have revealed that immune infiltration plays a vital part in the two. Nevertheless, more studies are required to confirm the relationship between immune infiltration and related differentially expressed gene regulation.
    OBJECTIVE: To screen the differentially expressed genes with prognostic significance between myelodysplastic syndrome and acute myeloid leukemia by bioinformatics analysis and explore the possible roles and mechanisms among these differentially expressed genes and immune infiltration mechanisms in the occurrence and progression of diseases. 
    METHODS: The differentially expressed genes were screened for bioinformatics analysis using the GEO datasets, and analyzed by DO, GO, KEGG and GSEA. The TCGA prognostic database was used to plot the K-M curves of differentially expressed genes and receiver operating characteristic curve analysis was applied to evaluate the clinical diagnostic performance. Finally, CIBERSORT analysis was used to intuitively demonstrate the correlation between critical prognostic genes and the distribution of immuno-infiltrated cells. RT-qPCR was employed to detect peripheral blood samples from healthy controls, myelodysplastic syndrome and acute myeloid leukemia patients so as to verify the crucial genes preliminarily.
    RESULTS AND CONCLUSION: (1) A total of 150 differentially expressed genes were obtained between myelodysplastic syndrome and acute myeloid leukemia, among which 16 genes were up-regulated and 134 were down-regulated. (2) The results of DO, GO, KEGG and GSEA analysis suggested that differentially expressed genes might promote the development of myelodysplastic syndrome to acute myeloid leukemia by regulating the immune response. CIBERSORT revealed the differences in immune infiltration between myelodysplastic syndrome and acute myeloid leukemia. The distribution of CD4+ T cells, monocytes, neutrophils and M1 macrophages decreased in acute myeloid leukemia patients. In contrast, the distribution of inflammatory suppressor cells M2 macrophages increased, suggesting that it may be related to the immunosuppression of acute myeloid leukemia. (3) K-M curve and receiver operating characteristic curve analysis of 150 differentially expressed genes screened out four genes relevant to immunity and prognosis with good diagnostic performance: MANSC1, FLT3, BMX and CXCR2. (4) The results of RT-qPCR exhibited that MANSC1, BMX and CXCR2 were low expressed, while FLT3 was highly expressed in acute myeloid leukemia patients. These findings verify that the differential expression of MANSC1, FLT3, BMX and CXCR2 in patients with myelodysplastic syndrome and acute myeloid leukemia is not only significantly correlated with the prognosis of patients but may also affect the occurrence and development of myelodysplastic syndrome and acute myeloid leukemia by regulating the immune infiltration of patients. They can be used as potential biomarkers and therapeutic targets of the transformation from myelodysplastic syndrome to acute myeloid leukemia, providing a new direction for clinical diagnosis and treatment of the transformation of myelodysplastic syndrome. 
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    Construction of nomogram and validation of clinical prediction model for high-quality blastocyst formation in patients with unexplained infertility
    Zhou Chao, Jiang Yueyuan, Yu Guangyu, Yu Chunmei
    2024, 28 (13):  2090-2097.  doi: 10.12307/2024.130
    Abstract ( 199 )   PDF (1090KB) ( 19 )   Save
    BACKGROUND: Unexplained infertility is associated with a higher abortion rate and lower fertilization rate, implantation rate, clinical pregnancy rate and cumulative live birth rate. It is urgent to establish a clinical prediction model related to infertility of unknown cause to solve the problems of clinical prognosis and individualized medical services, and finally achieve the purpose of increasing the cumulative live birth rate of patients with infertility of unknown cause.
    OBJECTIVE: To construct and verify the prediction model of high-quality blastocyst formation in patients with unexplained infertility during in vitro fertilization.
    METHODS: A total of 419 patients with unknown infertility who underwent in vitro fertilization in the Assisted Reproduction Department of Changzhou Maternal and Child Health Care Hospital from March 2017 to June 2022 were retrospectively analyzed, including 317 patients with high-quality blastocysts and 102 patients without high-quality blastocysts. A prediction model was established and used as the model group. The model group was sampled 1 000 times by the Bootstrap method as the validation group. Firstly, the univariate analysis was used to screen the influencing factors of high-quality blastocyst formation of unknown infertility, and the best matching factors were selected by the least absolute shrinkage and selection operator (LASSO) algorithm. Multiple factors were included in the progressive Logistic regression to find out the independent influencing factors and draw a column graph. Finally, the subject working curve, calibration curve, clinical decision curve and clinical impact curve were used to verify the differentiation and accuracy of the prediction model as well as the clinical application efficiency.
    RESULTS AND CONCLUSION: (1) Univariate analysis of the factors influencing the formation of high-quality blastocyst of unknown infertility were age, insemination method, antimullerian hormone level, basal follicle-stimulating hormone level, basal luteinizing hormone level, human chorionic gonadotropin injection day follicle-stimulating hormone level, human chorionic gonadotropin day estradiol level, progesterone level on human chorionic gonadotropin day, the number of high-quality cleavage embryo (day 3) and the number of blastocyst formation (P < 0.05). (2) The best matching factors further screened by LASSO regression were age, insemination method, antimullerian hormone level, basal luteinizing hormone level, human chorionic gonadotropin injection day follicle-stimulating hormone level, human chorionic gonadotropin day estradiol level, the number of high-quality cleavage embryo (day 3) and the number of blastocyst formation (P < 0.05). Multifactor stepwise Logistic regression results showed that independent influencing factors on the formation of high-quality blastocysts for unexplained infertility were age, insemination method, antimullerian hormone level, the number of high-quality cleavage embryo (day 3), and the number of blastocyst formation. (3) Receiver operating characteristic curve exhibited that the area under the curve was 0.880(0.834, 0.926) in the model group and 0.889(0.859, 0.918) in the validation group. It showed that the prediction model had good differentiation. The average absolute error of the calibration curve was 0.036, indicating that the model had good accuracy. The Hosmer-Lemeshow test showed that there was no statistical difference between the prediction probability of blastocyst formation and the actual probability of blastocyst formation (P > 0.05). The clinical decision curve and clinical impact curve showed that the model group and the validation group had the maximum clinical net benefit when the threshold probability value was (0.16-0.96) and (0.08-0.93), respectively, and had better clinical application efficacy within the threshold probability range. These findings concluded that age, insemination method, antimullerian hormone, the number of high-quality cleavage embryos (day 3), and the number of blastocyst formation were independent factors influencing the formation of the fine blastocyst in patients with unexplained infertility. The clinical prediction model constructed by these factors has good clinical prediction value and clinical application efficiency and can provide a basis for clinical prognosis and intervention as well as the formulation of individual medical programs.
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    Factors affecting differentiation of tendon stem/progenitor cells
    Lu Jingwei, Lyu Kexin, Jiang Li, Chen Yixuan, Shi Houyin, Li Sen
    2024, 28 (13):  2098-2104.  doi: 10.12307/2024.142
    Abstract ( 208 )   PDF (888KB) ( 27 )   Save
    BACKGROUND: Tendinopathy is a musculoskeletal disorder characterized by pain and decreased mobility, with pathological changes of disturbed collagen and hyperplasia of the vasculature. Tendinopathy tends to occur in athletes, physical workers, and the elderly. One of the mechanisms of tendinopathy is the “failed healing response”, and part of what causes the failed healing response is the erroneous differentiation of tendon stem/progenitor cells. 
    OBJECTIVE: By reviewing the relevant literature, we introduce the characteristics of tendon stem/progenitor cells, summarize the factors that affect the differentiation of tendon stem/progenitor cells to tendon cells and those that lead to mis-differentiation of tendon stem/progenitor cells (differentiation to adipocytes, osteocytes and chondrocytes), and also describe the limitations of tendon stem/progenitor cells in clinical applications.  
    METHODS: PubMed and Web of Science databases were searched for the terms “tendon stem/progenitor cells, tendinopathy, tendon injury, differentiation”. The relevant literature was screened by reading and 109 articles were included for the analysis of the results.
    RESULTS AND CONCLUSION: (1) Tendon stem/progenitor cells are a type of stem cells that can spontaneously differentiate into tendons and have the ability to self-renew, clone, and multi-differentiate. Various external conditions acting on tendon stem/progenitor cells can lead them to differentiate in diverse directions. The specific factors that regulate the fate of tendon stem/progenitor cells are not known with certainty. When stem cell renewal and differentiation in tendons becomes abnormal, it can lead to failure of tendon healing and consequently to tendinopathy. (2) Aging, changes in extracellular matrix composition, excessive mechanical stimulation, prostaglandin E2 and interleukin-6 as well as interleukin-10 and some systemic diseases may be important in regulating the mis-differentiation of tendon stem/progenitor cells. (3) Possible favorable factors that promote the differentiation of tendon stem/progenitor cells to tenocytes are: some growth factors and cytokines, moderate mechanical stimulation and topography of the extracellular matrix, low oxygen tension, drugs, and several transcriptional genes and proteins. (4) The most desirable therapeutic tools are the regulation of endogenous tendon stem/progenitor cells or the stimulation of endogenous tendon stem/progenitor cell proliferation and differentiation by exogenous tendon stem/progenitor cells. (5) Understanding the factors that regulate mis-differentiation of tendon stem/progenitor cells may provide insight into the pathogenesis of tendinopathy and identify therapeutic targets. Elaborating on the induction of tendon stem/progenitor cell differentiation into tendons could facilitate their use in tissue engineering. 
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    Effect of exosomes and the preconditioning method on pulp regeneration
    Yang Runze, Wang Wei, Chen San, Zhou Xuedong, Wu Jiayuan
    2024, 28 (13):  2105-2113.  doi: 10.12307/2024.108
    Abstract ( 207 )   PDF (1212KB) ( 53 )   Save
    BACKGROUND: Existing studies have confirmed that exosomes can effectively promote pulp regeneration. However, the biological functions and properties of exosomes from preconditioned sources can be significantly changed, which have different effects on cell proliferation, migration and odontogenic differentiation.
    OBJECTIVE: To discuss the application status of exosomes and their preconditioning methods in the field of pulp regeneration, and summarize the preconditioning methods that affect the function of exosomes, and explore the effect of exosomes and their preconditioning on pulp regeneration.
    METHODS: The relevant articles were searched in WanFang, CNKI, PubMed, and Web of Science databases from 2006 to 2022. The Chinese and English search terms were “exosomes, pulp regeneration; preconditioning method”. A total of 78 articles were included for analysis.

    RESULTS AND CONCLUSION: (1) Exosomes have the advantages of good biocompatibility, low immunogenicity and no cytotoxicity, and can induce the regeneration of pulp tissue by promoting stem cell tooth formation, neurogenesis and vascularization. (2) Exosomes derived from preconditioning can enhance the ability of tissue repair and regeneration and have a significant impact on the quality of regenerated dental pulp. (3) Currently, the preconditioning methods used in the field of dental pulp regeneration include inflammatory stimulation, hypoxia induction, conditioned medium and three-dimensional culture, and secreted exosomes can effectively improve the quality of regenerated dental pulp. Nevertheless, the specific effect and mechanism of different preconditioning methods on pulp regeneration need to be explored. 

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    Factors influencing allogeneic platelet concentrate in chronic refractory wounds
    Li Jiao, Li Xiaofeng, Li Jianping
    2024, 28 (13):  2114-2119.  doi: 10.12307/2024.155
    Abstract ( 178 )   PDF (906KB) ( 60 )   Save
    BACKGROUND: Clinical application of autologous platelet-rich plasma is limited by the condition of patients and by the quantity and activity of platelets. Transfusable platelet component blood platelet concentrates, collected and prepared by blood collection and supply agencies, are widely available, can be standardized and are effective in the clinic. 
    OBJECTIVE: To review the factors influencing the preparation of platelet concentrate by the buffy-coat method for the treatment of chronic refractory wounds.
    METHODS: A computer search of the PubMed, CNKI, and WanFang databases for platelet-related articles from January 2000 to April 2023 was conducted using the Chinese and English search terms “allogeneic platelets(concentrate), buffy-coat method, skin ulcer, refractory wounds”. The titles and abstracts were screened, and the full text was reviewed, resulting in the inclusion of 51 articles that met the subject criteria. 
    RESULTS AND CONCLUSION: (1) Platelet concentrates supplied by blood collection and supply agencies are effective in the treatment of diabetic foot wounds, pressure sores, lower limb venous ulcers and other chronic refractory wounds: reducing the size of the ulcer and shortening the wound healing time, especially in the first two weeks, with significant healing-promoting effects. (2) The storage conditions of the raw material used to prepare platelet concentrates (whole blood overnight/buffy-coat overnight, shaking/resting (time)), centrifugation conditions (centrifugation parameters, canning method) and the structure of the blood bag can affect the concentration of platelet concentrates, indirectly affecting the efficacy of allogeneic platelet concentrates. (3) There are also some problems with studies of allogeneic platelet concentrates for the treatment of chronic refractory wounds, such as the sample size of the available evidence is small, the results lack the optimal preparation parameters, optimal dose and treatment regimen for platelet concentrates to promote healing of different wounds are unknown, so more randomized multicenter clinical studies are needed.
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    Influence of adipose tissue and its derivative on wound repair and vascularization
    Zhao Lixin, Tang Qingxi, Zhang Kezhong
    2024, 28 (13):  2120-2125.  doi: 10.12307/2024.153
    Abstract ( 186 )   PDF (847KB) ( 20 )   Save
    BACKGROUND: Vascularization is essential for wound healing and functional recovery during soft tissue repair. Adipose tissue is believed to be the body’s largest source of stem cells, and a number of different fat complexes have been developed for research and treatment. Its ability to promote angiogenesis and soft tissue repair has been extensively studied.
    OBJECTIVE: To review the progress of vascularization in soft tissue repair, and to summarize the preparation methods of adipose tissue and its derivative and their applications in vascularization and soft tissue repair. It is proven that adipose tissue and its derivative have excellent research value and clinical application prospects in vascular and soft tissue engineering.
    METHODS: PubMed, Web of Science and CNKI databases were used to search the related articles published from January 2010 to February 2023. The search terms were “soft tissue repair, wound healing, vascularization, angiogenesis, adipose tissue, stromal vascular fraction, adipose tissue-derived microvascular fragment, nanofat, adipose extracellular matrix/stromal vascular fraction gel” in Chinese and English. A small number of old classic literature was also included. An initial screening was performed by reading the titles and abstracts to exclude literature that was not relevant to the topic of the article, and 69 papers were finally included for the analysis of the result.
    RESULTS AND CONCLUSION: (1) Wound healing is an important physiological process, which mainly occurs when tissue is damaged, such as injury, surgery, burn, tumor, infection and vascular disease caused by tissue damage and defects. (2) Adequate vascularization of the wound site is essential for tissue repair, reconstruction of local homeostasis and functional recovery. (3) Adipose tissue is believed to be the body’s largest pool of stem cells and a number of different fat components have been used for research and treatment. (4) Due to its inherent composition and preparation advantages, adipose tissue will continue to play an important role in tissue engineering research and therapy.
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    Bibliometrics and visualization-based analysis of research landscape of exosomes and foresight
    Zheng Ying, Huang Kemin
    2024, 28 (13):  2126-2132.  doi: 10.12307/2023.296
    Abstract ( 225 )   PDF (2648KB) ( 31 )   Save
    BACKGROUND: Exosomes have become one of the frontiers of biomedical research in recent years because of their promising applications in clinical diagnosis and therapy.  
    OBJECTIVE: To reveal the basic research and technology development trend of exosomes at home and abroad, and provide suggestions for related scientific and technological research and development in China.
    METHODS: This paper presented a quantitative analysis of the overall research and development situation, the frontier hotspots and future development trends in this research area by means of bibliometric methods and tools.  
    RESULTS AND CONCLUSION: The research and development hotspots in this area have entered a high growth phase with the application of new tools and methods. Many new technologies and products are facing clinical applications and markets with the development of a large number of clinical translation experiments. Comparative analysis at home and abroad reveals that China has made more remarkable achievements in basic research in this field, but there are still weaknesses in the possession of independent intellectual property rights, clinical technology translation, and regulatory system standardization and management, which need to be further strengthened in the future.
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