1.Proportions of memory T cells and expression of their associated cytokines in lymph nodes of mice infected with Echinococcus multilocularis
Yinshi LI ; Duolikun ADILAI ; Bingqing DENG ; Ainiwaer ABIDAN ; Sheng SUN ; Wenying XIAO ; Conghui GE ; Na TANG ; Jing LI ; Hui WANG ; Tao JIANG ; Chuanshan ZHANG
Chinese Journal of Schistosomiasis Control 2025;37(2):136-143
Objective To investigate the effects of Echinococcus multilocularis infection on levels of memory T (Tm) cells and their subsets in lymph nodes of mice at different stages of infection, so as to provide new insights into immunotherapy for alveolarechinococcosis. MethodsTwenty-four C57BL/6J mice aged 6 to 9 weeks were randomly divided into the infection group and the control group, of 12 mice in each group. Mice in the infection group were administered with 3 000 E. multilocularis protoscoleces via portal venous injection, while animals in the control group were administered with an equal volume of physiological saline. Three mice from each group were sacrificed 4, 12 weeks and 24 weeks post-infection, and lymph nodes were sampled and stained with hematoxylin and eosin (HE) to investigate the histopathological changes of mouse lymph nodes in the infection group. The expression and localization of T lymphocyte surface markers CD3, CD4, and CD8 were observed in mouse lymph nodes using immunohistochemical staining. In addition, lymphocyte suspensions were prepared from mouse lymph nodes in both groups at different time points post-infection, and the levels of Tm cell subsets and their secreted cytokines were detected using flow cytometry. Results HE staining showed diffuse structural alterations in the subcapsular cortical and paracortical regions of mouse lymph nodes in the infection group 4 weeks post-infection with E. multilocularis. Immunohistochemical staining detected CD3, CD4 and CD8 expression in mouse lymph nodes in both groups. Flow cytometry revealed higher proportions of CD4+ Tm cells [(55.3 ± 4.8)% vs. (38.8 ± 6.1)%; t = -4.259, P < 0.05] and CD4+ tissue-resident Tm (Trm) cells [(57.7 ± 3.7)% vs. (34.1 ± 11.2)%; t = -3.990, P < 0.05] in mouse lymph nodes in the infection group than in the control group 4 weeks post-infection, and higher proportions of CD4+ Tm cells [(34.6 ± 3.2)% vs. (23.3 ± 7.5)%; t = -2.764, P < 0.05] and CD4+ Trm cells [(44.0 ± 1.9)% vs. (31.2 ± 1.5)%; t = -4.039, P < 0.05] in mouse lymph nodes in the infection group than in the control group 24 weeks post-infection. The proportions of CD8+ Tm cells were higher in the infection group than in the control group 4 weeks [(56.8 ± 2.7)% vs. (43.9 ± 5.2)%; t = -4.416, P < 0.01] and 12 weeks post-infection [(25.4 ± 2.7)% vs. (12.0 ± 2.6)%; t = -2.552, P < 0.05], while the proportions of tumor necrosis factor (TNF)-α+ CD4+ T cells [(15.7 ± 5.0)% vs. (49.4 ± 6.4)%; t = 7.150, P < 0.01], TNF-α+CD8+ T cells [(20.7 ± 5.5)% vs. (57.5 ± 8.4)%; t = -6.694, P < 0.01], and TNF-α+ CD8+ Tm cells [7.0% (1.0%) vs. 31.0% (11.0%); Z = -2.236, P < 0.05] were lower in the infection group than in the control group 24 weeks post-infection. Conclusions Tm cells levels are consistently increased in lymph nodes of mice at different stages of E. multilocularis infection, with Trm cells as the predominantly elevated subset. The impaired capacity of CD8+ Tm cells to secrete the effector molecule TNF-α in mouse lymph nodes at the late-stage infection may facilitate chronic parasitism of E. multilocularis.
2.Protective effect of secreted secretomes derived from three-dimensional cultured mesenchymal stem cells on mouse hippocampal neurons with seawater immersion and trauma
Xiao-Juan LAN ; Sheng-Qiang XIE ; Bing-Xian WANG ; Wen-Jing MA ; Qiao-Mi CHEN ; Gang CHENG ; Wen-Ying LYU ; Jian-Ning ZHANG
Medical Journal of Chinese People's Liberation Army 2025;50(3):324-331
Objective To investigate the protective effects of secretomes released by three-dimensional cultured mesenchymal stem cells(MSCs)on neurons subjected to seawater immersion(SW)and stretch injury(SI),and to provide new insights into neuronal repair following SW combined with traumatic brain injury(TBI).Methods MSCs were cultured using the hanging drop method,and the conditioned medium(CM)containing MSCs secretomes was collected.A cellular model combining SW with SI was established using mouse hippocampal neuronal cells(HT22 cells).HT22 cells were randomly assigned to five groups:control,SI,SI+SW,SI+CM,and SI+SW+CM groups.Cell viability was assessed using the CCK-8 assay,apoptosis rate was measured by flow cytometry,cell migration ability was evaluated by scratch assay,and the expression levels of apoptosis-related proteins Bcl-2 and Bcl-2-associated protein(Bax),and ferroptosis-related proteins long-chain acyl-CoA synthetase 4(ACSL4)and cyclooxygenase-2(COX-2)were detected by Western blotting.Results Immersion in 15%seawater for 12 h significantly decreased HT22 cell viability(P<0.05).The CCK-8 assay indicated that cell viability in both the SI and SI+SW groups was significantly lower than that in control group after 12 h of treatment(P<0.05).Treatment with CM containing MSCs secretomes significantly increased cell viability in SI+CM group compared to SI group(P<0.0001),and in SI+SW+CM group compared to SI+SW group(P<0.001).Flow cytometry results revealed that the apoptosis rate in SI and SI+SW groups was significantly higher than that in control group(P<0.05 or P<0.001),while in SI+CM group was lower than that in SI group(P<0.05),and in SI+SW+CM group was lower than that in SI+SW group(P<0.05).Western blotting showed that compared to control group,SI and SI+SW groups exhibited reduced Bcl-2 expression level(P<0.01 or P<0.0001)and increased expression levels of Bax,ACSL4,and COX-2(P<0.01 or P<0.0001).Compared to SI group,the SI+CM group displayed increased Bcl-2 expression level(P<0.05)and decreased expression levels of Bax,ACSL4,and COX-2(P<0.05).Compared to SI+SW group,SI+SW+CM group exhibited increased Bcl-2 expression level(P<0.01)and decreased expression levels of Bax,ACSL4,and COX-2(P<0.01 or P<0.001).Scratch assay results demonstrated that at both 12 h and 24 h,the cell migration rate in SI and SI+SW groups was significantly lower than that in control group(P<0.01 or P<0.0001),while the migration rate in SI+CM group was significantly higher than that in SI group(P<0.0001 or P<0.01),and the migration rate in SI+SW+CM group was significantly higher than that in SI+SW group(P<0.0001).Conclusion Secretomes derived from MSCs cultured using the hanging drop method can alleviate neuronal damage caused by SW and TBI,potentially offering a therapeutic approach for SW combined with TBI.
3.Identification of GSK3 family and regulatory effects of brassinolide on growth and development of Nardostachys jatamansi.
Yu-Yan LEI ; Zheng MA ; Jing WEI ; Wen-Bing LI ; Ying LI ; Zheng-Ming YANG ; Shao-Shan ZHANG ; Jing-Qiu FENG ; Hua-Chun SHENG ; Yuan LIU
China Journal of Chinese Materia Medica 2025;50(2):395-403
This study identified 8 members including NjBIN2 of the GSK3 family in Nardostachys jatamansi by bioinformatics analysis. Moreover, the phylogenetic tree revealed that the GKS3 family members of N. jatamansi had a close relationship with those of Arabidopsis. RT-qPCR results showed that NjBIN2 presented a tissue-specific expression pattern with the highest expression in roots, suggesting that NjBIN2 played a role in root growth and development. In addition, the application of epibrassinolide or the brassinosteroid(BR) synthesis inhibitor(brassinazole) altered the expression pattern of NjBIN2 and influenced the photomorphogenesis(cotyledon opening) and root development of N. jatamansi, which provided direct evidence about the functions of NjBIN2. In conclusion, this study highlights the roles of BIN2 in regulating the growth and development of N. jatamansi by analyzing the expression pattern and biological function of NjBIN2. It not only enriches the understanding about the regulatory mechanism of the growth and development of N. jatamansi but also provides a theoretical basis and potential gene targets for molecular breeding of N. jatamansi with improved quality in the future.
Brassinosteroids/metabolism*
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Steroids, Heterocyclic/metabolism*
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Gene Expression Regulation, Plant/drug effects*
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Plant Proteins/metabolism*
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Phylogeny
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Nardostachys/metabolism*
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Plant Growth Regulators/pharmacology*
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Plant Roots/drug effects*
4.Research progress in pharmacological activities and pharmacokinetics of geniposidic acid.
Zi-Wei LI ; Sheng-Lan QI ; Qing-Guang ZHANG ; Ling CHEN ; Jing HU ; Guang-Bo GE ; Feng HUANG
China Journal of Chinese Materia Medica 2025;50(13):3679-3691
Geniposidic acid(GA), a natural iridoid, exists in the roots, stems, leaves, flowers, bark, fruits, and seeds of medicinal plants of Rubiaceae, Eucommiaceae, and Plantaginaceae. Modern pharmacological studies have revealed that GA has multiple pharmacological activities, including organ-protective, anti-inflammatory, antioxidative, anti-osteoporosis, anti-neurodegenerative, and anti-cardiovascular effects. GA can enhance cell/organism defenses by upregulating key anti-inflammatory and antioxidant cytokines, while downregulating key node proteins in pro-inflammatory signaling pathways such as AhR and TLR4/MyD88, thereby exerting pharmacological effects such as organ protection. Pharmacokinetic investigations have suggested that after oral administration, GA can be distributed in multiple organs(kidney, liver, heart, spleen, lung, etc.). In addition, the pharmacokinetic behavior of GA could be significantly altered under disease conditions, as demonstrated by a marked increase in systematic exposure. This article comprehensively summarizes the reported pharmacological activities and mechanisms and systematically analyzes the pharmacokinetic characteristics and key parameters of GA, with the aim of providing a theoretical basis and scientific reference for the precise clinical application of GA-related Chinese patent medicines, as well as for the investigation and development of innovative drugs based on GA.
Humans
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Drugs, Chinese Herbal/chemistry*
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Animals
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Iridoid Glucosides/chemistry*
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Plants, Medicinal/chemistry*
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Anti-Inflammatory Agents/pharmacology*
5.Biomedical Data in China: Policy, Accumulation, Platform Construction, and Applications.
Jing-Chen ZHANG ; Jing-Wen SUN ; Xiao-Meng LIU ; Jin-Yan LIU ; Wei LUO ; Sheng-Fa ZHANG ; Wei ZHOU
Chinese Medical Sciences Journal 2025;40(1):9-17
Biomedical data is surging due to technological innovations and integration of multidisciplinary data, posing challenges to data management. This article summarizes the policies, data collection efforts, platform construction, and applications of biomedical data in China, aiming to identify key issues and needs, enhance the capacity-building of platform construction, unleash the value of data, and leverage the advantages of China's vast amount of data.
China
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Humans
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Biomedical Research
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Data Management
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Data Collection
6.Thalassemia screening and genotype analysis in 488 sperm donors in Chongqing Human Sperm Bank
Weiqiong SHENG ; Jing HUANG ; Tianfeng ZHANG ; Yi ZHANG ; Hua HUANG ; Hong WANG ; Guoqing JIA ; Chengjiang KANG
Journal of Chongqing Medical University 2025;50(4):511-515
Objective:To investigate the carriage of the thalassemia gene in sperm donors in Chongqing,China by analyzing the thalas-semia screening and gene mutation types of sperm donors in Chongqing Human Sperm Bank(CHSB),and to provide a reference for thalassemia screening in sperm donors in Chongqing.Methods:Blood samples were collected from 488 sperm donors in CHSB(donors with all tests qualified during the quarantine)from December 2021 to August 2024.A complete blood count analysis was performed to measure the mean corpuscular volume(MCV)and mean corpuscular hemoglobin(MCH).Polymerase chain reaction(PCR)/agarose gel electrophoresis was used to detect deletional α-thalassemia mutations.PCR and DNA reverse dot blot hybridization were used to detect 3 common non-deletional α-thalassemia mutations and 17 common β-thalassemia mutations.The test results were then analyzed.Results:Among the 488 sperm donors aged 25.64±4.95 years,15(3.07%,all Han nationality)tested positive for thalassemia.This carriage rate was lower than that of the thalassemia population in Chongqing(9.24%).Among the 15 thalassemia carriers,13 had deletional α-gene mutations(-α3.7 genotype in 11 cases[73.33%],-α4.2 genotype in 1 case[6.67%],and-SEA genotype in 1 case[6.67%]),and 2 had non-deletional α-gene mutations(both with the WSM heterozygous mutation).There was only 1 case of abnor-mal MCV and MCH2 indicators among thalassemia carriers(screening positive rate was 6.67%).Conclusion:Thalassemia screening in sperm donors in CHSB reveals a low thalassemia carriage rate,reflecting geographic(predominantly Sichuan and Chongqing ori-gins)and demographic(healthy men of childbearing potential)characteristics.Given the significant impact of thalassemia on the off-spring conceived through sperm donation,it is recommended to conduct thalassemia gene screening among sperm donors in human sperm banks.
7.The effect of rutaecarpine on improving fatty liver and osteoporosis in MAFLD mice
Yu-hao ZHANG ; Yi-ning LI ; Xin-hai JIANG ; Wei-zhi WANG ; Shun-wang LI ; Ren SHENG ; Li-juan LEI ; Yu-yan ZHANG ; Jing-rui WANG ; Xin-wei WEI ; Yan-ni XU ; Yan LIN ; Lin TANG ; Shu-yi SI
Acta Pharmaceutica Sinica 2025;60(1):141-149
Metabolic-associated fatty liver disease (MAFLD) and osteoporosis (OP) are two very common metabolic diseases. A growing body of experimental evidence supports a pathophysiological link between MAFLD and OP. MAFLD is often associated with the development of OP. Rutaecarpine (RUT) is one of the main active components of Chinese medicine Euodiae Fructus. Our previous studies have demonstrated that RUT has lipid-lowering, anti-inflammatory and anti-atherosclerotic effects, and can improve the OP of rats. However, whether RUT can improve both fatty liver and OP symptoms of MAFLD mice at the same time remains to be investigated. In this study, we used C57BL/6 mice fed a high-fat diet (HFD) for 4 months to construct a MAFLD model, and gave the mice a low dose (5 mg·kg-1) and a high dose (15 mg·kg-1) of RUT by gavage for 4 weeks. The effects of RUT on liver steatosis and bone metabolism were then evaluated at the end of the experiment [this experiment was approved by the Experimental Animal Ethics Committee of Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences (approval number: IMB-20190124D303)]. The results showed that RUT treatment significantly reduced hepatic steatosis and lipid accumulation, and significantly reduced bone loss and promoted bone formation. In summary, this study shows that RUT has an effect of improving fatty liver and OP in MAFLD mice.
8.Knowledge, attitude and practice investigation of severe fever with thrombocytopenia syndrome among residents in epidemic areas in Qingdao
Lin LI ; Jingmin XIE ; Xiaowen WANG ; Xiaojuan GONG ; Xiaodong ZHANG ; Xinlei BI ; Changmin SHENG ; Jing JIA
Chinese Journal of Experimental and Clinical Virology 2025;39(2):162-168
Objective:To understand the knowledge, attitudes, and behaviors of residents in areas endemic for severe fever with thrombocytopenia syndrome (SFTS) and the factors influencing these aspects, to provide a scientific basis for the prevention and control of SFTS among residents in endemic areas.Methods:A multi-stage sampling method was used to select subjects for a questionnaire survey, and the collected data were analyzed and compared.Results:The awareness rate of SFTS-related knowledge among residents in the endemic areas of Qingdao was 14.38%, while the awareness rate of tick-related knowledge was 69.78%. The mean score for protective attitudes among residents in the endemic areas was 24.50±4.09, and the mean score for protective behaviors was 11.20±2.76. Among residents of different ages, occupations, and educational levels, comparisons of SFTS-related knowledge awareness rates ( χ2=293.553, P<0.001; χ2=495.216, P<0.001; χ2=677.272, P<0.001), tick-related knowledge awareness rates ( χ2=29.986, P<0.001; χ2=23.239, P=0.002; χ2=41.003, P<0.001), protective attitude scores ( H=52.094, P<0.001; H=33.446, P<0.001; H=81.715, P<0.001), and protective behavior scores ( H=52.344, P<0.001; H=28.465, P<0.001; H=67.061, P<0.001) showed statistically significant differences. Comparisons of these same factors among residents of different genders showed no statistically significant differences ( χ2=1.159, P=0.282; χ2=0.298, P=0.585; t=-0.808, P=0.419; t=-1.229, P=0.219). Comparisons of tick-related knowledge awareness rates among residents with different frequencies of outdoor work showed no statistically significant differences ( χ2=5.367, P=0.147); however, comparisons of SFTS-related knowledge awareness rates, protective attitude scores, and protective behavior scores among residents with different frequencies of outdoor work showed statistically significant differences ( χ2=117.010, P<0.001; F=4.127, P=0.006; H=56.280, P<0.001). Multivariate linear regression analysis revealed that older age, lower educational level, and less frequent outdoor work were associated with lower protective behavior scores ( t=-3.479, P<0.001; t=4.861, P<0.001; t=-7.753, P<0.001). Conclusions:Residents in the epidemic areas of SFTS have a low level of knowledge, attitude and practice. Factors influencing protective behaviors include age, occupation, educational level, and frequency of outdoor work. Farmers are a group that requires particular attention.
9.Establishment of RT-RPA/RAA-based detection method for four henipa viruses
Wenjun HE ; Yuanyuan GUO ; Sheng ZHANG ; Mengjie YANG ; Jing ZHANG ; Wenwen LEI ; Juan SONG ; Guizhen WU
Chinese Journal of Experimental and Clinical Virology 2025;39(4):502-509
Objective:To establish a rapid and accurate duplex real-time fluorescent reverse transcription-recombinase polymerase amplification/recombinase-aided amplification(RT-RPA/RAA)detection method for identification and differentiation of Nipah virus(NiV),Langya virus(LayV),Mojiang virus(MoJV),and Cedar virus(CedV).Methods:First,specific primers and probes were designed targeting the conserved L gene regions of NiV and LayV,as well as the conserved N gene regions of MoJV and CedV,respectively. The four viruses were divided into two groups for duplex detection. Subsequently,the optimal primer and probe combinations were screened by comparing the amplification efficiency of different primer pair combinations(F1/R1,F1/R2,F2/R1,F2/R2). The reaction temperature was optimized through temperature gradient settings from 37 ℃ to 42 ℃,and the amounts of primers and probes were optimized to establish the duplex real-time fluorescent RT-RPA/RAA detection system. Finally,the detection performance was evaluated through specificity,sensitivity,and stability tests,as well as clinical sample validation.Results:The selected primer pairs(NiV primer pair F2/R1,LayV primer pair F1/R1,MoJV primer pair F2/R2,and CedV primer pair F2/R2)all demonstrated optimal amplification efficiency when combined with their corresponding probes. The optimal annealing temperature was 39 ℃,and the minimum detection limit was 101-102 copies/μl. The method could effectively distinguish target viruses from other non-target viruses,and repeated experiments showed good stability( R2> 0.90). Additionally,detection results for Malaysian NiV strains and various clinical samples were consistent with the Taqman multiplex qRT-PCR method. Conclusion:The duplex real-time fluorescent RT-RPA/RAA detection method successfully established in this study can rapidly and accurately identify and differentiate four important henipavirus-like viruses:NiV,LayV,MoJV,and CedV. It features simple operation,rapid reaction,high specificity,and good stability,providing an effective molecular detection tool for rapid field diagnosis,surveillance,and control of these zoonotic viruses.
10.Role of CCL2/CCR2 signaling pathway in electroacupuncture-induced reduction of spinal cord injury in rats
Xiang WANG ; Jianzhong HUO ; Wei FAN ; Jing GAO ; Yangyang SHENG ; Jie ZHANG ; Zhaoyu ZHANG
Chinese Journal of Anesthesiology 2025;45(5):574-580
Objective:To evaluate the role of the CC chemokine ligand 2/CC chemokine receptor 2 (CCL2/CCR2) signaling pathway in electroacupuncture (EA)-induced reduction of spinal cord injury (SCI) in rats.Methods:Sixty clean-grade healthy adult female Sprague-Dawley rats, weighing 210-250 g, were divided into 5 groups ( n=12 each) using a random number table method: sham operation group (group S), group SCI, SCI+ Anti-CCL2 group (group SCI+ A), SCI+ EA group (group SCI+ EA), and spinal cord injury+ EA+ rCCL2 group (group SCI+ EA+ R). The SCI model was established using the Allen method in anesthetized animals. Group S only underwent spinous process and laminectomy without damaging the spinal cord. In SCI+ A group, CCL2 neutralizing antibody 50 μg/kg was intrathecally injected at 0, 3 and 6 days after successful development of the SCI model. On the 7th day after the successful development of the SCI model, Jiaji, Dazhui and Mingmen acupoints were stimulated with a depth of 2 mm, voltage of 12-15 mV and frequency of 2 Hz for 30 min once a day for 7 consecutive days in SCI+ EA group. In SCI+ EA+ R group, recombinant rat CCL2 2.5 μg/kg was intrathecally injected at the site of injury at 0, 3 and 6 days after successful development of the SCI model, and the remaining treatments were similar to those in SCI+ EA group. At 1 day before developing the model, 0, 3, 7, 14, 21 and 28 days after developing the model, the mechanical paw withdraw threshold (MWT) and thermal paw withdrawal latency (TWL) were measured, and the motor function was assessed by BBB score. The rats were sacrificed after the final behavioral testing, and their spinal cord tissues were obtained for determination of the expression of CCL2 and CCR2 protein and mRNA (by Western blot or quantitative real-time polymerase chain reaction), the expression of GFAP (by immunofluorescence), contents of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 (by enzyme-linked immunosorbent assay) and for examination of the pathological changes (using HE staining). Results:Compared with S group, the MWT and BBB scores were significantly decreased and the TWL was shortened at each time point after developing the model, the expression of CCL2 and CCR2 protein and mRNA and GFAP was up-regulated, and the contents of TNF-α, IL-1β and IL-6 were increased in SCI group ( P<0.05). Compared with SCI group, the MWT and BBB scores were significantly increased, and the TWL was prolonged at 7 days after developing the model in SCI+ A group, the MWT and BBB scores were significantly increased, and the TWL was prolonged at 14 days after developing the model in SCI+ EA group, and the expression of CCL2 and CCR2 protein and mRNA and GFAP was significantly down-regulated, and the contents of TNF-α, IL-1β and IL-6 were decreased in SCI+ A and SCI+ EA groups ( P<0.05). Compared with SCI+ EA group, the MWT and BBB scores were significantly decreased at 14 days after developing the model, the TWL was shortened, the expression of CCL2 and CCR2 protein and mRNA and GFAP was up-regulated, and the contents of TNF-α, IL-1β and IL-6 were increased in SCI+ EA+ R group ( P<0.05). Compared with SCI+ A and SCI+ EA groups, the histopathological injury were significantly attenuated in SCI group, and the histopathological injury was aggravated in SCI+ EA+ R group. Conclusions:The CCL2/CCR2 signaling pathway is involved in the process by which EA reduces SCI, and the mechanism is related to the inhibition of astrocyte activation, thereby reducing the inflammatory response.

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