ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

ObjectiveMagnetoencephalography (MEG), a non-invasive neuroimaging technique, meticulously captures the magnetic fields emanating from brain electrical activity. Compared with MEG based on superconducting quantum interference devices (SQUID), MEG based on optically pump magnetometer (OPM) has the advantages of higher sensitivity, better spatial resolution and lower cost. However, most of the current studies are clinical studies, and there is a lack of animal studies on MEG based on OPM technology. Pain, a multifaceted sensory and emotional phenomenon, induces intricate alterations in brain activity, exhibiting notable sex differences. Despite clinical revelations of pain-related neuronal activity through MEG, specific properties remain elusive, and comprehensive laboratory studies on pain-associated brain activity alterations are lacking. The aim of this study was to investigate the effects of inflammatory pain (induced by Complete Freund’s Adjuvant (CFA)) on brain activity in a rat model using the MEG technique, to analysis changes in brain activity during pain perception, and to explore sex differences in pain-related MEG signaling. MethodsThis study utilized adult male and female Sprague-Dawley rats. Inflammatory pain was induced via intraplantar injection of CFA (100 μl, 50% in saline) in the left hind paw, with control groups receiving saline. Pain behavior was assessed using von Frey filaments at baseline and 1 h post-injection. For MEG recording, anesthetized rats had an OPM positioned on their head within a magnetic shield, undergoing two 15-minute sessions: a 5-minute baseline followed by a 10-minute mechanical stimulation phase. Data analysis included artifact removal and time-frequency analysis of spontaneous brain activity using accumulated spectrograms, generating spectrograms focused on the 4-30 Hz frequency range. ResultsMEG recordings in anesthetized rats during resting states and hind paw mechanical stimulation were compared, before and after saline/CFA injections. Mechanical stimulation elevated alpha activity in both male and female rats pre- and post-saline/CFA injections. Saline/CFA injections augmented average power in both sexes compared to pre-injection states. Remarkably, female rats exhibited higher average spectral power 1 h after CFA injection than after saline injection during resting states. Furthermore, despite comparable pain thresholds measured by classical pain behavioral tests post-CFA treatment, female rats displayed higher average power than males in the resting state after CFA injection. ConclusionThese results imply an enhanced perception of inflammatory pain in female rats compared to their male counterparts. Our study exhibits sex differences in alpha activities following CFA injection, highlighting heightened brain alpha activity in female rats during acute inflammatory pain in the resting state. Our study provides a method for OPM-based MEG recordings to be used to study brain activity in anaesthetized animals. In addition, the findings of this study contribute to a deeper understanding of pain-related neural activity and pain sex differences.

Implementing the reform of outpatient mutual-aid is a crucial measure to improve China's medical security system.Based on the policy diffusion theory,we took 228 prefecture-level cities across the country as the research objects and used the event history method to analyze the diffusion mode and influencing factors of the general outpatient pooling policy for employees'basic medical insurance,and divided into three types:the type of reducing the stock of Medical Savings Accounts(MSA),the welfare increment type I and type II for separate analysis.We found that the diffusion of the general outpatient pooling policy presented an"S"-shaped curve over time,and was mainly horizontally diffused in space,with a neighboring effect and a regional demonstration effect.The type of reducing the stock of MSA policy diffuses horizontally;the welfare increment type I is mainly horizontal diffusion,supplemented by vertical diffusion;the welfare increment type II is not affected by the pressure from higher-level governments,and mainly adopts the horizontal diffusion mode.In terms of the influencing factors of policy diffusion,the diffusion of the general outpatient pooling policy was affected by multiple factors such as policy issues,legitimacy,and government capacity.The diffusion of the type of reducing the stock of MSA policy was not affected by the pressure from higher-level governments,but was influenced by policy issues and government capacity.The diffusion of the welfare increment type I policy was most affected by policy issues,and the welfare increment type II was most affected by government capacity.

Aim To investigate the effect of cordycepin(COR)on gentamicin(GEN)-induced nephrotoxicity and the molecular mechanism of inhibiting oxidative stress and ferroptosis induced by GEN.Methods The oral SD rats were divided into a control group,GEN group,and GEN+COR group.Following the success-ful setting up of the animal model,the serum creatinine(CR)and urea nitrogen(BUN)levels of rats were measured,and renal tissue injury was assessed using HE staining.In addition,the contents of malondialde-hyde and glutathione in kidney tissues of SD rats in each group were detected,and the expressions of fer-roptosis markers GPX4 and SLC7A11 were analyzed by Western blot.Results Compared with the control group,CR and BUN in GEN-stimulated group signifi-cantly increased(P＜0.01),and the level of CR and BUN was effectively reduced after 50 mg·kg-1 COR oral administration.HE results also showed that COR could alleviate the kidney tissue damage caused by GEN.COR could reverse the increase of malondialde-hyde level and the decrease of glutathione level caused by GEN in rat kidney tissue,and COR could restore the decrease of GPX4 and SLC7A11 protein levels induced by GEN.Conclusion COR can reduce GEN-induced kidney injury by inhibiting oxidative stress and ferrop-tosis.

Objective:To apply the Timed Up and Go Test(TUGT)to investigate the correlation between motor function, emotional state, cognitive function, and sleep quality among elderly individuals in the Chinese community.Methods:A cross-sectional study was conducted, involving 739 subjects aged 60 to 90 years, who were randomly recruited from December 2021 to August 2023 across Beijing, Tianjin, Zhejiang, Guangdong, and Hainan Provinces in China.Basic demographic information was collected, and the TUGT was utilized to assess motor function.Based on the TUGT time(t), the subjects were divided into three groups: normal motor function group, mild motor abnormality group, and significant motor abnormality group.Cognitive function was evaluated using the Chinese Revised Mini-Mental State Examination(MMSE), while the Patient Health Questionnaire Depression Scale(PHQ-9)was employed to measure the degree of depression.Additionally, the Epworth Sleepiness Scale(ESS)was used to assess excessive daytime sleepiness.The correlation between subjects' motor function and their cognitive abilities, mood, and sleep was subsequently analyzed.Results:Systolic blood pressure, heart rate, PHQ-9, MMSE, and ESS scores were identified as significant factors influencing TUGT time.Specifically, TUGT time was positively correlated with PHQ-9 and ESS scores, while exhibiting negative correlations with systolic blood pressure, heart rate, and MMSE scores.Additionally, TUGT time was negatively correlated with the MMSE subcomponents of orientation, immediate memory, and verbal ability.All observed differences were statistically significant(all P<0.05).Logistic regression analysis indicated that an increase in the PHQ-9 score was associated with an odds ratio( OR)of 1.099(95% CI: 1.045-1.155, P<0.001)(mild motor abnormality group)and 1.150(95% CI: 1.066-1.242, P<0.001)(Significant motor abnormality group).Additionally, a reduction in the MMSE score was observed, with an OR of 0.939(95% CI: 0.886-0.995, P<0.001)(mild motor abnormality group)and 0.793(95% CI: 0.729-0.862, P<0.001)(Significant motor abnormality group).Furthermore, an increase in the ESS score was noted, with ORs of 1.139(95% CI: 1.094-1.186, P<0.001)(mild motor abnormality group)and 1.203(95% CI: 1.132-1.279, P<0.001)(Significant motor abnormality group).These findings suggest that these variables are independently related to decreased motor function. Conclusions:Depression, cognitive impairment, and excessive daytime sleepiness are independent risk factors for motor dysfunction among elderly individuals in community settings.The Timed Up and Go Test TUGT can be utilized for the early screening of motor function decline in this population.

Objective:To apply the Timed Up and Go Test(TUGT)to investigate the correlation between motor function, emotional state, cognitive function, and sleep quality among elderly individuals in the Chinese community.Methods:A cross-sectional study was conducted, involving 739 subjects aged 60 to 90 years, who were randomly recruited from December 2021 to August 2023 across Beijing, Tianjin, Zhejiang, Guangdong, and Hainan Provinces in China.Basic demographic information was collected, and the TUGT was utilized to assess motor function.Based on the TUGT time(t), the subjects were divided into three groups: normal motor function group, mild motor abnormality group, and significant motor abnormality group.Cognitive function was evaluated using the Chinese Revised Mini-Mental State Examination(MMSE), while the Patient Health Questionnaire Depression Scale(PHQ-9)was employed to measure the degree of depression.Additionally, the Epworth Sleepiness Scale(ESS)was used to assess excessive daytime sleepiness.The correlation between subjects' motor function and their cognitive abilities, mood, and sleep was subsequently analyzed.Results:Systolic blood pressure, heart rate, PHQ-9, MMSE, and ESS scores were identified as significant factors influencing TUGT time.Specifically, TUGT time was positively correlated with PHQ-9 and ESS scores, while exhibiting negative correlations with systolic blood pressure, heart rate, and MMSE scores.Additionally, TUGT time was negatively correlated with the MMSE subcomponents of orientation, immediate memory, and verbal ability.All observed differences were statistically significant(all P<0.05).Logistic regression analysis indicated that an increase in the PHQ-9 score was associated with an odds ratio( OR)of 1.099(95% CI: 1.045-1.155, P<0.001)(mild motor abnormality group)and 1.150(95% CI: 1.066-1.242, P<0.001)(Significant motor abnormality group).Additionally, a reduction in the MMSE score was observed, with an OR of 0.939(95% CI: 0.886-0.995, P<0.001)(mild motor abnormality group)and 0.793(95% CI: 0.729-0.862, P<0.001)(Significant motor abnormality group).Furthermore, an increase in the ESS score was noted, with ORs of 1.139(95% CI: 1.094-1.186, P<0.001)(mild motor abnormality group)and 1.203(95% CI: 1.132-1.279, P<0.001)(Significant motor abnormality group).These findings suggest that these variables are independently related to decreased motor function. Conclusions:Depression, cognitive impairment, and excessive daytime sleepiness are independent risk factors for motor dysfunction among elderly individuals in community settings.The Timed Up and Go Test TUGT can be utilized for the early screening of motor function decline in this population.

AIM To investigate the protective effect of Sanfeng Tongqiao Dropping Pills against house dust mite(HDM)-induced allergic asthma in mice.METHODS Compared to the intact BALB/c mice in the blank control group,the BALB/c mice randomly assigned into the model group,the dexamethasone group(0.67 mg/kg),and the low-dose,medium-dose,and high-dose Sanfeng Tongqiao Dropping Pills groups(15,30 and 60 mg/kg),were induced into acute allergic asthma models via weekly intraperitoneal sensitization with 0.1 mL HDM solution(0.5 mg/mL)for three weeks followed by three consecutive daily intranasal challenges with 10 μL HDM solution(2.5 mg/mL)starting in the third week.The drug administered continuously 7 days after the last excitation.The mice had their airway reactive Penh value detected,their pulmonary pathological changes observed by HE staining,their blood eosinophils(EOS)counted,their Th2 cytokines in lung tissue and serum IgE levels detected by ELISA,and their number of peripheral blood mononuclear cells(PBMC)and pulmonary Th2 cells detected by flow cytometry.Chronic allergic asthma was induced in grouped BALB/c mice through repeated intranasal challenges with 10 μL HDM solution(2.5 mg/mL)administered five times weekly for five consecutive weeks.Drug treatment continued for 14 days following the final challenge.After the final treatment,the mice had their pulmonary pathological changes observed by HE staining,and their levels of Th2 cytokines in B ALF and lung tissue and serum IgE detected by ELISA.RESULTS Compared to the blank control group,the acute allergic asthma model group exhibited increases in Penh value,EOS count and IgE level in serum,IL-4 and IL-5 levels in lung tissue(P＜0.01);obvious pulmonary inflammatory cells infiltration,and thickened airway wall;and increase in pulmonary number of Th2 cells(P＜0.01).Compared to the model group,the groups intervened with Sanfeng Tongqiao Dropping Pills demonstrated decreased Penh value,serum EOS count,IgE level and IL-5 level in lung tissue(P＜0.05,P＜0.01);reduced pulmonary inflammatory infiltration and alleviated airway wall thickening;and decreased number of pulmonary Th2 cells.Compared to the blank group,the chronic allergic asthma model group showed obvious pulmonary inflammatory infiltration and airway wall thickening;and increased EOS count and IgE level in serum,IL-4 and IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).Compared to the model group,the groups intervened with either medium-dose or high-dose Sanfeng Tongqiao Dropping Pills demonstrated reduced pulmonary inflammatory infiltration;and decreased serum EOS count,IgE level,IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).CONCLUSION Sanfeng Tongqiao Dropping Pills reduce Th2 cells in peripheral blood and lung tissue,suppress type 2 inflammation,and thereby alleviate allergic asthma.

Objective To investigate the effect of G9a histone methyltransferase inhibitor BIX01294 on the proliferation of vascular smooth muscle cell(VSMC)and its underlying mechanism.Methods Twelve SD rats were divided into the control group and the treatment group,with 6 rats in each group.The rats were anesthetized by intraperitoneal injection with sodium pentobarbital,then exposed the common carotid artery and the internal and external carotid artery after disinfection and skin preparation,ligated the distal end of the internal and external jugular veins,clamped the proximal end of the common carotid artery,cut the external carotid artery,inserted the balloon catheter,blocked the blood flow by compression,and ligated the proximal end of the external carotid artery after withdrawing the balloon.The common carotid artery clamping state of rats in the treatment group was maintained after withdrawing the balloon,then restored the blood flow after perfusion with 100 μmol/L BIX01294 solution for 30 seconds;rats in the control group were restored the blood flow after perfusion with PBS for 30 seconds.VSMC were divided into the normal group and the BIX01294 groups.Cells in the normal group were cultured in low glucose medium containing 2%fetal bovine serum,and cells in the BIX01294 groups were co-treated with 2.5,5.0,7.5 and 10.0 μmol/L BIX01294 on the basis of the normal group,respectively.CCK-8 assay and EDU assay were used to detect the cell activity and proliferative ability respectively,to screen appropriate concentration of BIX01294.The cell apoptosis was detected by TUNEL assay between the normal group and the BIX01294 group,and the control group and the treatment group.Western blot was used to detect the expression levels of autophagy-and apoptosis-related proteins in the normal group and BIX01294 group.Results Compared with 0 μmol/L,the activity and proliferative ability of VSMC decreased in a concentration-dependent manner after treatment with BIX01294 at different concentrations(P<0.05).Compared with the normal group,the apoptosis level of VSMC in the BIX01294 group was increased(P<0.05),the expression of VSMC autophagy-and apoptosis-related proteins of LC3Ⅰ/Ⅱ and Bax were up-regulated(P<0.05),and the expression of Bcl-2 and p62 proteins were down-regulated(P<0.05).In vivo test results showed that BIX01294 local perfusion aggravated the apoptosis of carotid VSMC.Conclusion BIX01294 activates VSMC autophagy and apoptosis and inhibits its proliferation.

Implementing the reform of outpatient mutual-aid is a crucial measure to improve China's medical security system.Based on the policy diffusion theory,we took 228 prefecture-level cities across the country as the research objects and used the event history method to analyze the diffusion mode and influencing factors of the general outpatient pooling policy for employees'basic medical insurance,and divided into three types:the type of reducing the stock of Medical Savings Accounts(MSA),the welfare increment type I and type II for separate analysis.We found that the diffusion of the general outpatient pooling policy presented an"S"-shaped curve over time,and was mainly horizontally diffused in space,with a neighboring effect and a regional demonstration effect.The type of reducing the stock of MSA policy diffuses horizontally;the welfare increment type I is mainly horizontal diffusion,supplemented by vertical diffusion;the welfare increment type II is not affected by the pressure from higher-level governments,and mainly adopts the horizontal diffusion mode.In terms of the influencing factors of policy diffusion,the diffusion of the general outpatient pooling policy was affected by multiple factors such as policy issues,legitimacy,and government capacity.The diffusion of the type of reducing the stock of MSA policy was not affected by the pressure from higher-level governments,but was influenced by policy issues and government capacity.The diffusion of the welfare increment type I policy was most affected by policy issues,and the welfare increment type II was most affected by government capacity.