1.Action mechanism of intrathecal transplantation of human umbilical cord mesenchymal stem cell-derived exosomes for repair of spinal cord injury under neuroendoscopy
Yitong ZHENG ; Yongxin WANG ; Wen LIU ; AMUJITE ; Hu QIN
Chinese Journal of Tissue Engineering Research 2025;29(36):7743-7751
BACKGROUND:Human umbilical cord mesenchymal stem cell-derived exosomes were found to be effective in promoting neural repair in spinal cord injury.OBJECTIVE:To investigate whether exosomes derived from human umbilical cord mesenchymal stem cells are able to attenuate neuroinflammation and promote recovery of motor function by promoting polarization of microglia toward the M2 type.METHODS:Totally 48 SD rats were randomly divided into a sham operation group,a model group,and an exosome group(n=16 per group).A rat spinal cord injury model was established using the modified Allen method.The exosome group was injected with 20 μL of human umbilical cord mesenchymal stem cell-derived exosomes intrathecally via neuroendoscopy 24 hours after injury.At 3,7,14,and 21 days after modeling,the recovery of the motor function of the hind limbs of the rats was assessed by BBB scoring method combined with Rivlin's slant plate test.The damage of spinal cord tissues was detected by using hematoxylin-eosin staining and Nissl staining.The expression levels of brain-derived neurotrophic factor and vascular endothelial growth factor A proteins were detected by western blot assay.The expression proportion of M1-type markers(inducible nitric oxide synthase)and M2 markers(arginase-1)in the spinal cord tissues was detected by immunofluorescence method.qRT-PCR and western blot assay were used to detect the expression levels of inducible nitric oxide synthase and arginase-1 in spinal cord tissues.ELISA was utilized to detect the levels of pro-inflammatory factors(tumor necrosis factor α,interleukin 1β,and interleukin 6)and anti-inflammatory factors(interleukin 10)levels in spinal cord tissues.RESULTS AND CONCLUSION:(1)At 3,7,and 14 days postoperatively,the BBB scores of the exosome group were better than those of the model group(P<0.05).The angles of the Rivlin slanting plate experiments of the exosome group were significantly higher than those of the model group at 7 and 14 days postoperatively(P<0.05).The results of hematoxylin-eosin staining and Nissl staining indicated that the spinal cord tissues and nerve injuries of the exosome group were reduced in comparison with those of the model group,and the levels of brain-derived neurotrophic factor and vascular endothelial growth factor A in spinal cord tissues of the exosome group were higher than those in the model group at 7 days postoperatively(P<0.05).(2)Immunofluorescence experiments showed that the number of inducible nitric oxide synthase-positive microglial cells in the lesion area of the exosome group was significantly reduced and the level of Arg1-positive microglial cells increased in the lesion area of the exosome group compared with the model group at 7 days postoperatively(P<0.05).qRT-PCR and western blot assay also confirmed the results of immunofluorescence experiments.(3)The secretion of pro-inflammatory factors tumor necrosis factor α,interleukin 1β,and interleukin 6 in spinal cord tissues of the exosome group was reduced compared with the model group(P<0.05),whereas the secretion of the inflammation-suppressing factor interleukin 10 was increased compared with the model group(P<0.05).These findings conclude that human umbilical cord mesenchymal stem cell-derived exosomes could promote the polarization of microglial cells from the M1 to the M2 type and decrease the release of pro-inflammatory factors,thereby reducing the secondary damage of neuroinflammation in spinal cord injury.
2.miR-30a-3p exacerbates nonalcoholic fatty liver disease in mice via in-hibiting hepatocyte autophagy and promoting lipid deposition
Diao CHEN ; Yongxin CHEN ; Guiting WANG ; Birong YUAN ; Longkun HU ; Tian ZHANG ; Bing GUO
Chinese Journal of Pathophysiology 2025;41(5):953-964
AIM:To explore whether microRNA-30a-3p(miR-30a-3p)is involved in the pathogenesis of non-alcoholic fatty liver disease(NAFLD)by regulating autophagy and promoting lipid deposition.METHODS:Eight-week-old C57BL/6 mice were randomly divided into a normal control group and a high-fat diet(HFD)group.Mice in the HFD group were fed with 60%high fat diet for 10 weeks to induce the NAFLD phenotype.Some mice were injected with adeno-virus overexpressing miR-30a-3p via the tail vein and subsequently fed with high-fat diet for 4 weeks.Glucose tolerance and insulin resistance tests were performed at the end of the treatments.In addition,the concentrations of hepatic alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglyceride(TG)and total cholesterol(TC)were mea-sured.Hematoxylin-eosin staining and oil red O staining were conducted to examine morphological changes and lipid depo-sition in the liver tissue.The expression levels of microtubule-associated protein light chain 3(LC3),autophagy-related protein 5(ATG5),beclin-1 and p62 were quantified through Western blot.In addition,NAFLD models were established in AML12 hepatocytes by incubating the cells with palmitic acid and oleic acid(PO).The AML12 cells were transfected with miR-30a-3p shRNA to knock down miR-30a-3p expression.The concentration levels of TG and TC after miR-30a-3p knockdown were measured by the kits.Nile red staining was performed to examine lipid droplet aggregation and dual fluo-rescent recombinant adenovirus Ad-mCherry-GFP-LC3B was transfected into AML12 cells to observe changes in autopha-gic flow.RESULTS:HFD-fed mice exhibited significant insulin resistance and reduced glucose tolerance,significant lip-id deposition in the liver tissue,coupled with increased hepatic ALT,AST,TG and TC levels.The expression levels of au-tophagy-related proteins LC3-Ⅱ,beclin-1,and ATG5 were decreased,while that of p62 was increased(P<0.01).More-over,miR-30a-3p overexpression significantly increased blood glucose and insulin resistance in HFD-fed mice.However,it aggravated lipid droplets deposition in liver tissue and enhanced hepatic TG,TC,AST and ALT levels.Western blot re-vealed that the expression levels of LC3-Ⅱ,beclin-1 and ATG5 were further reduced,while that of p62 was significantly in-creased(P<0.01).In vitro,we observed that the TG and TC levels,as well as lipid accumulation in PO-treated AML12 cells were increased significantly.Similarly,the expression levels of LC3-Ⅱ,beclin-1 and ATG5 were decreased,whereas that of p62 increased in PO-treated AML12 cells(P<0.01).Notably,knockdown of miR-30a-3p resulted in a significant reduction in the TG content in PO-treated AML12 cells and lipid droplet aggregation was significantly suppressed.Further-more,the expression of LC3-Ⅱ,beclin-1 and ATG5 proteins was increased,while that of p62 was decreased significantly and the autophagy flow was improved(P<0.01).CONCLUSION:The miR-30a-3p exacerbates hepatic lipid deposi-tion,inducing severe hepatic steatosis and liver damage,to promote the occurrence and development of NAFLD in mice.Mechanistically,its effects involve inhibition of hepatic autophagy level.
3.miR-30a-3p exacerbates nonalcoholic fatty liver disease in mice via in-hibiting hepatocyte autophagy and promoting lipid deposition
Diao CHEN ; Yongxin CHEN ; Guiting WANG ; Birong YUAN ; Longkun HU ; Tian ZHANG ; Bing GUO
Chinese Journal of Pathophysiology 2025;41(5):953-964
AIM:To explore whether microRNA-30a-3p(miR-30a-3p)is involved in the pathogenesis of non-alcoholic fatty liver disease(NAFLD)by regulating autophagy and promoting lipid deposition.METHODS:Eight-week-old C57BL/6 mice were randomly divided into a normal control group and a high-fat diet(HFD)group.Mice in the HFD group were fed with 60%high fat diet for 10 weeks to induce the NAFLD phenotype.Some mice were injected with adeno-virus overexpressing miR-30a-3p via the tail vein and subsequently fed with high-fat diet for 4 weeks.Glucose tolerance and insulin resistance tests were performed at the end of the treatments.In addition,the concentrations of hepatic alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglyceride(TG)and total cholesterol(TC)were mea-sured.Hematoxylin-eosin staining and oil red O staining were conducted to examine morphological changes and lipid depo-sition in the liver tissue.The expression levels of microtubule-associated protein light chain 3(LC3),autophagy-related protein 5(ATG5),beclin-1 and p62 were quantified through Western blot.In addition,NAFLD models were established in AML12 hepatocytes by incubating the cells with palmitic acid and oleic acid(PO).The AML12 cells were transfected with miR-30a-3p shRNA to knock down miR-30a-3p expression.The concentration levels of TG and TC after miR-30a-3p knockdown were measured by the kits.Nile red staining was performed to examine lipid droplet aggregation and dual fluo-rescent recombinant adenovirus Ad-mCherry-GFP-LC3B was transfected into AML12 cells to observe changes in autopha-gic flow.RESULTS:HFD-fed mice exhibited significant insulin resistance and reduced glucose tolerance,significant lip-id deposition in the liver tissue,coupled with increased hepatic ALT,AST,TG and TC levels.The expression levels of au-tophagy-related proteins LC3-Ⅱ,beclin-1,and ATG5 were decreased,while that of p62 was increased(P<0.01).More-over,miR-30a-3p overexpression significantly increased blood glucose and insulin resistance in HFD-fed mice.However,it aggravated lipid droplets deposition in liver tissue and enhanced hepatic TG,TC,AST and ALT levels.Western blot re-vealed that the expression levels of LC3-Ⅱ,beclin-1 and ATG5 were further reduced,while that of p62 was significantly in-creased(P<0.01).In vitro,we observed that the TG and TC levels,as well as lipid accumulation in PO-treated AML12 cells were increased significantly.Similarly,the expression levels of LC3-Ⅱ,beclin-1 and ATG5 were decreased,whereas that of p62 increased in PO-treated AML12 cells(P<0.01).Notably,knockdown of miR-30a-3p resulted in a significant reduction in the TG content in PO-treated AML12 cells and lipid droplet aggregation was significantly suppressed.Further-more,the expression of LC3-Ⅱ,beclin-1 and ATG5 proteins was increased,while that of p62 was decreased significantly and the autophagy flow was improved(P<0.01).CONCLUSION:The miR-30a-3p exacerbates hepatic lipid deposi-tion,inducing severe hepatic steatosis and liver damage,to promote the occurrence and development of NAFLD in mice.Mechanistically,its effects involve inhibition of hepatic autophagy level.
4.Action mechanism of intrathecal transplantation of human umbilical cord mesenchymal stem cell-derived exosomes for repair of spinal cord injury under neuroendoscopy
Yitong ZHENG ; Yongxin WANG ; Wen LIU ; AMUJITE ; Hu QIN
Chinese Journal of Tissue Engineering Research 2025;29(36):7743-7751
BACKGROUND:Human umbilical cord mesenchymal stem cell-derived exosomes were found to be effective in promoting neural repair in spinal cord injury.OBJECTIVE:To investigate whether exosomes derived from human umbilical cord mesenchymal stem cells are able to attenuate neuroinflammation and promote recovery of motor function by promoting polarization of microglia toward the M2 type.METHODS:Totally 48 SD rats were randomly divided into a sham operation group,a model group,and an exosome group(n=16 per group).A rat spinal cord injury model was established using the modified Allen method.The exosome group was injected with 20 μL of human umbilical cord mesenchymal stem cell-derived exosomes intrathecally via neuroendoscopy 24 hours after injury.At 3,7,14,and 21 days after modeling,the recovery of the motor function of the hind limbs of the rats was assessed by BBB scoring method combined with Rivlin's slant plate test.The damage of spinal cord tissues was detected by using hematoxylin-eosin staining and Nissl staining.The expression levels of brain-derived neurotrophic factor and vascular endothelial growth factor A proteins were detected by western blot assay.The expression proportion of M1-type markers(inducible nitric oxide synthase)and M2 markers(arginase-1)in the spinal cord tissues was detected by immunofluorescence method.qRT-PCR and western blot assay were used to detect the expression levels of inducible nitric oxide synthase and arginase-1 in spinal cord tissues.ELISA was utilized to detect the levels of pro-inflammatory factors(tumor necrosis factor α,interleukin 1β,and interleukin 6)and anti-inflammatory factors(interleukin 10)levels in spinal cord tissues.RESULTS AND CONCLUSION:(1)At 3,7,and 14 days postoperatively,the BBB scores of the exosome group were better than those of the model group(P<0.05).The angles of the Rivlin slanting plate experiments of the exosome group were significantly higher than those of the model group at 7 and 14 days postoperatively(P<0.05).The results of hematoxylin-eosin staining and Nissl staining indicated that the spinal cord tissues and nerve injuries of the exosome group were reduced in comparison with those of the model group,and the levels of brain-derived neurotrophic factor and vascular endothelial growth factor A in spinal cord tissues of the exosome group were higher than those in the model group at 7 days postoperatively(P<0.05).(2)Immunofluorescence experiments showed that the number of inducible nitric oxide synthase-positive microglial cells in the lesion area of the exosome group was significantly reduced and the level of Arg1-positive microglial cells increased in the lesion area of the exosome group compared with the model group at 7 days postoperatively(P<0.05).qRT-PCR and western blot assay also confirmed the results of immunofluorescence experiments.(3)The secretion of pro-inflammatory factors tumor necrosis factor α,interleukin 1β,and interleukin 6 in spinal cord tissues of the exosome group was reduced compared with the model group(P<0.05),whereas the secretion of the inflammation-suppressing factor interleukin 10 was increased compared with the model group(P<0.05).These findings conclude that human umbilical cord mesenchymal stem cell-derived exosomes could promote the polarization of microglial cells from the M1 to the M2 type and decrease the release of pro-inflammatory factors,thereby reducing the secondary damage of neuroinflammation in spinal cord injury.
5.Clinical guidelines for the diagnosis and treatment of osteoporotic thoracolumbar vertebral fracture with kyphotic deformity in the elderly (version 2024)
Jian CHEN ; Qingqing LI ; Jun GU ; Zhiyi HU ; Shujie ZHAO ; Zhenfei HUANG ; Tao JIANG ; Wei ZHOU ; Xiaojian CAO ; Yongxin REN ; Weihua CAI ; Lipeng YU ; Tao SUI ; Qian WANG ; Pengyu TANG ; Mengyuan WU ; Weihu MA ; Xuhua LU ; Hongjian LIU ; Zhongmin ZHANG ; Xiaozhong ZHOU ; Baorong HE ; Kainan LI ; Tengbo YU ; Xiaodong GUO ; Yongxiang WANG ; Yong HAI ; Jiangang SHI ; Baoshan XU ; Weishi LI ; Jinglong YAN ; Guangzhi NING ; Yongfei GUO ; Zhijun QIAO ; Feng ZHANG ; Fubing WANG ; Fuyang CHEN ; Yan JIA ; Xiaohua ZHOU ; Yuhui PENG ; Jin FAN ; Guoyong YIN
Chinese Journal of Trauma 2024;40(11):961-973
The incidence of osteoporotic thoracolumbar vertebral fracture (OTLVF) in the elderly is gradually increasing. The kyphotic deformity caused by various factors has become an important characteristic of OTLVF and has received increasing attention. Its clinical manifestations include pain, delayed nerve damage, sagittal imbalance, etc. Currently, the definition and diagnosis of OTLVF with kyphotic deformity in the elderly are still unclear. Although there are many treatment options, they are controversial. Existing guidelines or consensuses pay little attention to this type of fracture with kyphotic deformity. To this end, the Lumbar Education Working Group of the Spine Branch of the Chinese Medicine Education Association and Editorial Committee of Chinese Journal of Trauma organized the experts in the relevant fields to jointly develop Clinical guidelines for the diagnosis and treatment of osteoporotic thoracolumbar vertebral fractures with kyphotic deformity in the elderly ( version 2024), based on evidence-based medical advancements and the principles of scientificity, practicality, and advanced nature, which provided 18 recommendations to standardize the clinical diagnosis and treatment.
6.Real-time Detection Method for Motion Artifact of Photoplethysmography Signals Based on Decision Trees
Linqi HU ; Yulin ZHANG ; Yongxin CHOU ; Haiping YANG ; Xiao HE
Chinese Journal of Medical Instrumentation 2024;48(3):285-292
PPG(photoplethysmography)holds significant application value in wearable and intelligent health devices.However,during the acquisition process,PPG signals can generate motion artifacts due to inevitable coupling motion,which diminishes signal quality.In response to the challenge of real-time detection of motion artifacts in PPG signals,this study analyzed the generation and significant features of PPG signal interference.Seven features were extracted from the pulse interval data,and those exhibiting notable changes were filtered using the dual-sample Kolmogorov-Smirnov test.The real-time detection of motion artifacts in PPG signals was ultimately based on decision trees.In the experimental phase,PPG signal data from 20 college students were collected to formulate the experimental dataset.The experimental results demonstrate that the proposed method achieves an average accuracy of(94.07±1.14)%,outperforming commonly used motion artifact detection algorithms in terms of accuracy and real-time performance.
7.Selection and Weight Determination of Diagnostic Items for Qi Stagnation Syndrome Based on Delphi Method and Analytic Hierarchy Process
Xuan ZHOU ; Ge FANG ; Qingyu MA ; Xiaojuan LI ; Yongxin LI ; Zhixi HU ; Xiantao LI ; Jiaxu CHEN
Journal of Traditional Chinese Medicine 2024;65(21):2211-2216
ObjectiveUsing the Delphi method and analytic hierarchy process (AHP) to screen diagnostic items for qi stagnation syndrome and determine their weights, providing a reference for the development of a diagnostic scale of qi stagnation syndrome. MethodsLiterature related to qi stagnation syndrome were screened from databases including CNKI, Wanfang, VIP, SinoMed (from inception to October 31, 2020). Through systematic review of literature and expert discussions, the information on the four examinations of traditional Chinese medicine were organized and an item pool was constructed. The Delphi method was used to screen the item indicators, while the AHP was employed to determine their weights. Statistical methods such as mean value, full score ratio, rank sum, unimportant percentage, and coefficient of variation were used for item screening, with the weights calculated by AHP serving as the item weights. ResultsA total of 235 articles and books were included for analysis, resulting in an item pool of 16 items. After three rounds of expert consultation, a total of 84 valid questionnaires were collected, with a total expert enthusiasm coefficient of 99% and authority coefficient of 0.86, 0.84, 0.83, respectively, and the coordination coefficients were 0.45, 0.49, and 0.29, respectively. Through the statistics analysis, 8 diagnosis items were screened out, including distension (stuffi-ness) or distending pain or scurrying pain, wiry pulse, depressed emotions, frequent sighing, deep and wiry pulse, irritability, pale red tongue, and thin white coating. The AHP showed that the order of weights of the first-level indicators from high to low was clinical symptoms, pulse manifestation, and tongue manifestation; the order of weights of the second-level indicators from high to low was distension (stuffiness) or distending pain or scurrying pain, wiry pulse, depressed emotions, frequent sighing, deep and wiry pulse, irritability, pale red tongue, and thin white coating. ConclusionBy applying the Delphi method and AHP to analyze and evaluate the diagnostic items for qi stagnation syndrome, key diagnostic items were screened and their weights determined, laying the foundation for the development of a diagnostic scale for qi stagnation syndrome.
8.Advances in the application of non-contrast CT radiomics and machine learning in the diagnosis and treat-ment process of hypertensive Intracerebral hemorrhage
Yanwen JIANG ; Hu QIN ; Zhaofu LENG ; Aikel-Amu PAZILIYA ; Yongxin WANG
Chinese Journal of Nervous and Mental Diseases 2023;49(10):609-614
Hypertensive intracerebral hemorrhage(HICH)is a disease with a rapid onset,rapid progression,high mortality rate,and long-term impact on the ability to function.Non-contrast agent-based CT(NCCT)is a common method for evaluating and identifying HICH.Recent radiomics in image processing and machine learning(ML)have enabled the extraction of high-dimensional feature information from medical images,which can be used to rapidly and accurately diagnose HICH and predict its course of disease.The paper describes the application of radiomics and ML techniques in HICH diagnosis and treatment,and identifies possible directions for future research.
9.Clinical guideline for spinal reconstruction of osteoporotic thoracolumbar fracture in elderly patients (version 2022)
Tao SUI ; Jian CHEN ; Zhenfei HUANG ; Zhiyi HU ; Weihua CAI ; Lipeng YU ; Xiaojian CAO ; Wei ZHOU ; Qingqing LI ; Jin FAN ; Qian WANG ; Pengyu TANG ; Shujie ZHAO ; Lin CHEN ; Zhiming CUI ; Wenyuan DING ; Shiqing FENG ; Xinmin FENG ; Yanzheng GAO ; Baorong HE ; Jianzhong HUO ; Haijun LI ; Jun LIU ; Fei LUO ; Chao MA ; Zhijun QIAO ; Qiang WANG ; Shouguo WANG ; Xiaotao WU ; Nanwei XU ; Jinglong YAN ; Zhaoming YE ; Feng YUAN ; Jishan YUAN ; Jie ZHAO ; Xiaozhong ZHOU ; Mengyuan WU ; Yongxin REN ; Guoyong YIN
Chinese Journal of Trauma 2022;38(12):1057-1066
Osteoporotic thoracolumbar fracture in the elderly will seriously reduce their quality of life and life expectancy. For osteoporotic thoracolumbar fracture in the elderly, spinal reconstruction is necessary, which should comprehensively consider factors such as the physical condition, fracture type, clinical characteristics and osteoporosis degree. While there lacks relevant clinical norms or guidelines on selection of spinal reconstruction strategies. In order to standardize the concept of spinal reconstruction for osteoporotic thoracolumbar fracture in the elderly, based on the principles of scientificity, practicality and progressiveness, the authors formulated the Clinical guideline for spinal reconstruction of osteoporotic thoracolumbar fracture in elderly patients ( version 2022), in which suggestions based on evidence of evidence-based medicine were put forward upon 10 important issues related to the fracture classification, non-operative treatment strategies and surgical treatment strategies in spinal reconstruction after osteoporosis thoracolumbar fracture in the elderly, hoping to provide a reference for clinical treatment.
10.Neuroimaging findings, pathological features and surgical efficacy of adult gangliogliomas
Muertizha MAMUTIJIANG ; Yongxin WANG ; Yandong FAN ; Hu QIN ; Qian MA
Chinese Journal of Neuromedicine 2022;21(9):929-933
Objective:To investigate the clinical characteristics, pathological features and surgical efficacy of adult gangliogliomas.Methods:Seven patients with gangliogliomas underwent surgical treatment in our hospital from August 2010 to December 2021, were chosen; their clinical data were retrospectively analyzed, and the CT and MRI manifestations and histopathological features of gangliogliomas and surgical efficacy of these patients were concluded.Results:In these 7 patients, solid-mass type was identified in 1 patient, cystic-solid mixed type in 4 patients, and diffuse infiltrating type in 2 patients. The solid part showed slightly low or equal density on CT, and mostly showed slightly low signal on T1, slightly high signal on T2, high or slightly high signal on T2 FLAIR, and equisignal or slightly low signal on DWI of MRI. Immunohistochemical staining showed that in the tumor specimens, glial cell components were positive for glial fibrillary acidic protein, oligodendrocyte transcription factor and S-100 protein, and ganglion cell components were positive for neuronal nuclear antigen and synaptophysin. Gross total resection was achieved in 5 patients, subtotal resection in 1 patient, and partial resection in 1 patient; gangliogliomas without total resection were diffuse infiltrative type. The median Karnofsky performance status scores were 80. There was no death during the follow-up period of 7 months-11 years.Conclusions:Adult gangliogliomas are mostly cystic and solid lesions with clear boundary on imaging, and the histopathological characteristics of mixed neoplastic ganglion cells and glial cells are the main basis for their diagnosis. The surgical efficacy is usually good, but the diffuse infiltrating type is difficult to achieve total resection.

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