Objective To explore the effect of aerobic exercise on sarcopenia in CT26 tumor mice through the gut microbiota-skeletal muscle axis and its potential mechanism.Methods Forty-eight SPF BALB/c male mice aged 8 weeks were randomly divided into a normal control group(group NC,n=12),a normal control+aerobic exercise group(group NA,n=12),a CT26 tumor-associated sarco-penia group(group TS,n=12)and a CT26 tumor-associated sarcopenia+aerobic exercise group(group TSA,n=12).After a 2-week adaptive exercise period,CT26 cell suspension(0.2 mL,1×107 cells/mL)was subcutaneously injected into the dorsal region of mice in group TS and group TSA,while group NC and group NA were inoculated with 0.2 mL normal saline.One day later,group NA and group TSA underwent a 4-week aerobic treadmill running intervention(14 m/min,60 min/day,6 days/week).If a mouse failed to complete the target intensity on a given day,its running speed was reduced by 20%the following day.Moreover,group NA and group TSA received no exercise interven-tion.General health status of all groups was monitored throughout the study.The forelimb gripping force,rotarod walking time and the cross-sectional area of gastrocnemius muscle fibers were measured.Moreover,the ultrastructural changes of gastrocnemius were observed by transmission electron micro-scope,while the cecum contents were observed by 16S rRNA sequencing.Meanwhile,the pathological morphological changes of small intestine wall were observed using HE,while the contents of tumor ne-crosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)and lipopoly saccharide(LPS)of serum and gastrocnemius muscle were determined by using ELISA.Moreover,small intestine zonula occludens-1(ZO-1),Occludin and muscle ring finger 1(MuRF1),muscle atrophy F-box(MAF-bx),Toll-like receptor 4(TLR4)/myeloid differentiation factor 8(MyD88)/nuclear transcription factor-κB(NF-κB)signal pathway related proteins expression levels of gastrocnemius muscle,as well as muscle ring finger 1(MuRF1)and muscle atrophy F-box(MAF-bx)were detected using Western blot.Results(1)Subcutaneous tumors were prominent in group TS.The forelimb gripforce,wet weight and cross-sectional area of gastrocnemius,and rotarod walking time in group TS at the 5th and 6th weekends were significantly lower than group NC(P<0.01),while the relative expression levels of MuRF1 and MAF-bx proteins in the gastrocnemius muscle in group TS were significantly higher than the latter group(P<0.05,P<0.01).Moreover,transmission electron microscopy revealed sparse and dis-organized muscle fiber arrangement in group TS,with visible Z-lines but indistinct M-lines,as well as shortened,poorly defined,and thin sarcomeres.(2)The group TSA exhibited significantly higher α-diversity indices(Shannon,Chao1,and Observed species)in gut microbiota compared to group TS(P<0.05,P<0.01,P<0.05),while the β-diversity differed to some extent between the two groups.At phylum level,the F/B ratio was lower in group NA than that group NC,with that of group TSA low-er than group TS.At genus level,the relative abundances of Muribaculaceae,Bacteroides,Roseburia,and Alistipes in group NA were higher than group NC,and those of Alloprevotellaand Helicobacter were significantly lower,while those of Muribaculaceae,Lachnospiraceae_NK4A136_group,Roseburia,Bacteroides,Lachnospiraceae_UCG-001 of group TSA were significantly higher than group TS.Nota-bly,Roseburia and Alistipes were the most significantly enriched genera in group TSA and group TS(LDA score>4 for both).(3)The inflammatory cells of the small intestine wall in group TSA signifi-cantly reduced than those in group TS(P<0.05).Moreover,the expression level of ZO-1 proteins in the small intestine wall were significantly higher in group NA than group NC,and that of ZO-1 and Occludin proteins in the small intestine wall was significantly higher in group TSA than those in group TS(P<0.05).(4)The group NA exhibited significantly lower LPS concentrations in the gastrocne-mius muscle compared to group NC(P<0.05).Similarly,the group TSA showed reduced LPS levels in both serum and gastrocnemius compared to group TS(P<0.05).The relative protein expression of p-IκBα and p-NF-κB p65 significantly decreased in group TSA compared to group TS(P<0.05).The group TS displayed elevated concentrations of pro-inflammatory cytokines(TNF-α,IL-1β)in serum and gastrocnemius compared to group NC(P<0.01,P<0.05).The group TSA demonstrated significantly lower levels of TNF-α and IL-6 in these tissues in comparison with group TS(P<0.05,P<0.01).Conclusion Aerobic exercise can improve gut microbiota α diversity,adjusting its composition,improv-ing intestinal mucosal barrier function,reducing the LPS-induced pro-inflammatory response,and delay-ing skeletal muscle atrophy.The underlying mechanism may involve down regulation of TLR4/MyD88/NF-κB signaling in skeletal muscle,which has a positive effect on Gut-Muscle Axis.

Objective To investigate the effect of aerobic exercise preconditioning on cardiac function in mice bearing subcutaneous transplantable tumors and explore the potential molecular mechanisms.Methods Twenty-four 6-week-old male BALB/c mice were selected.After an acclimation period,they were randomly assigned to a control group(C),a tumor group(M)and an exercise-preconditioning plus tumor group(EM),each of eight.The EM group underwent a 4-week aerobic exercise interven-tion.Meanwhile,the C group received 0.2 ml of physiological saline injected subcutaneously on the dorsum of the proximal left hind limb,while the M and EM groups were inoculated at the same site with 0.2 ml of a CT26.WT colon carcinoma cell suspension.Three weeks after inoculation,cardiac function was assessed by transthoracic echocardiography.Hearts were then harvested for hematoxylin-eo-sin staining to evaluate cardiomyocyte cross-sectional area(CSA),while Western blot was performed to determine the expression of proteins related to myocardial protein synthesis and degradation.Results(1)Compared with group C,group M exhibited significantly lower body weight and heart weight(P<0.05),and reduced left ventricular ejection fraction(LVEF),fractional shortening(FS)and stroke vol-ume(SV)(P<0.05),as well as cardiomyocyte CSA(P<0.01)and total mTOR(P<0.01),phosphory-lated mTOR(p-mTOR)(P<0.01)and the p-mTOR/mTOR ratio(P<0.05),but a significant increase in the expression of the muscle ring finger 1(MuRF-1)and the muscle atrophy F-box(MAFbx/Atrog-in-1)(P<0.01 and P<0.05,respectively).(2)Compared with group M,group EM showed significant-ly greater heart weight(P<0.05);increased left ventricular posterior wall thickness at end-diastole(LVPWd),LVEF,FS and SV(P<0.05),as well as the larger cardiomyocyte CSA and total mTOR,p-mTOR and the p-mTOR/mTOR ratio(P<0.05),but reduced MuRF-1 and Atrogin-1 expression(P<0.05).Conclusion Four weeks of aerobic exercise preconditioning ameliorated myocardial atrophy and improved systolic cardiac function in mice bearing subcutaneously transplanted CT26 tumors.Such beneficial effects may be associated with exercise-induced down regulation of the protein degradation mediators MuRF-1 and Atrogin-1 and up regulation of total mTOR and p-mTOR.

Objective To investigate the effects of aerobic exercise and different exercise habits on skeletal muscle function and the possible molecular mechanisms in colorectal cancer-loaded mice.Methods Thirty-five 5-week-old BABL/c male mice were acclimatized to feeding for 1 week and then divided randomly into the following groups:control(D),tumor(M),exercise preconditioning(QAM),lifetime exercise(AM),and exercise(HAM)groups(n=7 mice per group).Mice in the QAM and AM groups underwent aerobic exercise regimen 1 from weeks 2～6.At week 7,mice in the experimental groups received 0.2 mL of CT26 colon cancer cell suspension subcutaneously in the dorsal aspect of the left hind limb,while control mice received 0.2 mL of saline at the corresponding site.Mice in the AM and HAM groups were subjected to aerobic exercise regimen 2 for weeks 7～9.The general status and skeletal muscle mass and function were monitored in all mice throughout the experiments.After completion of the experiment,samples were collected and the cross-sectional area of gastrocnemius muscle fibers(CSA)was measured by hematoxylin and eosin staining and expression levels of proteins related to synthesis and catabolism of the gastrocnemius muscle were analyzed by Western Blot.Results(1)The weight ratio of the gastrocnemius muscle was significantly lower in mice in the M,QAM,and HAM groups compared with group D,and was significantly higher in AM mice compared with M,QAM,and HAM mice.(2)Grip strength,endurance,skeletal muscle circumference,and CSA were significantly lower in group D mice compared with the other groups,and was most enhanced in group HAM.Endurance and CSA were consistently enhanced in groups QAM,AM,and HAM.(3)Muscle RING-finger protein-1(MuRF1)expression levels were significantly lower in groups M,QAM,AM,and HAM than in group D,significantly lower in groups AM and HAM than in group M,and significantly lower in group HAM than in group QAM.(4)Fibronectin type Ⅲ domain-containing protein 5(FNDC5)expression levels were significantly lower in group M than in groups D and QAM.(5)Peroxisome proliferator-activated receptor gamma coactivator 1-alpha expression levels were significantly lower in the QAM and HAM groups compared with group M.(6)Expression levels of phospho(p)-AMP-activated protein kinase(AMPK)/AMPK and p-AMPK were significantly higher in group QAM than in groups D and M,p-AMPK expression significantly lower in groups AM and HAM was than in group QAM,and AMPK expression was significantly lower in groups QAM,AM,and HAM than in group D.Conclusions Exercise preconditioning and continuous aerobic exercise can improve skeletal muscle mass and function in CT26 colon cancer-loaded mice by activating AMPK phosphorylation to stimulate skeletal muscle secretion of FNDC5,thereby regulating the expression of MuRF1 protein.

Objective To explore the effect of aerobic exercise on sarcopenia in CT26 tumor mice through the gut microbiota-skeletal muscle axis and its potential mechanism.Methods Forty-eight SPF BALB/c male mice aged 8 weeks were randomly divided into a normal control group(group NC,n=12),a normal control+aerobic exercise group(group NA,n=12),a CT26 tumor-associated sarco-penia group(group TS,n=12)and a CT26 tumor-associated sarcopenia+aerobic exercise group(group TSA,n=12).After a 2-week adaptive exercise period,CT26 cell suspension(0.2 mL,1×107 cells/mL)was subcutaneously injected into the dorsal region of mice in group TS and group TSA,while group NC and group NA were inoculated with 0.2 mL normal saline.One day later,group NA and group TSA underwent a 4-week aerobic treadmill running intervention(14 m/min,60 min/day,6 days/week).If a mouse failed to complete the target intensity on a given day,its running speed was reduced by 20%the following day.Moreover,group NA and group TSA received no exercise interven-tion.General health status of all groups was monitored throughout the study.The forelimb gripping force,rotarod walking time and the cross-sectional area of gastrocnemius muscle fibers were measured.Moreover,the ultrastructural changes of gastrocnemius were observed by transmission electron micro-scope,while the cecum contents were observed by 16S rRNA sequencing.Meanwhile,the pathological morphological changes of small intestine wall were observed using HE,while the contents of tumor ne-crosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)and lipopoly saccharide(LPS)of serum and gastrocnemius muscle were determined by using ELISA.Moreover,small intestine zonula occludens-1(ZO-1),Occludin and muscle ring finger 1(MuRF1),muscle atrophy F-box(MAF-bx),Toll-like receptor 4(TLR4)/myeloid differentiation factor 8(MyD88)/nuclear transcription factor-κB(NF-κB)signal pathway related proteins expression levels of gastrocnemius muscle,as well as muscle ring finger 1(MuRF1)and muscle atrophy F-box(MAF-bx)were detected using Western blot.Results(1)Subcutaneous tumors were prominent in group TS.The forelimb gripforce,wet weight and cross-sectional area of gastrocnemius,and rotarod walking time in group TS at the 5th and 6th weekends were significantly lower than group NC(P<0.01),while the relative expression levels of MuRF1 and MAF-bx proteins in the gastrocnemius muscle in group TS were significantly higher than the latter group(P<0.05,P<0.01).Moreover,transmission electron microscopy revealed sparse and dis-organized muscle fiber arrangement in group TS,with visible Z-lines but indistinct M-lines,as well as shortened,poorly defined,and thin sarcomeres.(2)The group TSA exhibited significantly higher α-diversity indices(Shannon,Chao1,and Observed species)in gut microbiota compared to group TS(P<0.05,P<0.01,P<0.05),while the β-diversity differed to some extent between the two groups.At phylum level,the F/B ratio was lower in group NA than that group NC,with that of group TSA low-er than group TS.At genus level,the relative abundances of Muribaculaceae,Bacteroides,Roseburia,and Alistipes in group NA were higher than group NC,and those of Alloprevotellaand Helicobacter were significantly lower,while those of Muribaculaceae,Lachnospiraceae_NK4A136_group,Roseburia,Bacteroides,Lachnospiraceae_UCG-001 of group TSA were significantly higher than group TS.Nota-bly,Roseburia and Alistipes were the most significantly enriched genera in group TSA and group TS(LDA score>4 for both).(3)The inflammatory cells of the small intestine wall in group TSA signifi-cantly reduced than those in group TS(P<0.05).Moreover,the expression level of ZO-1 proteins in the small intestine wall were significantly higher in group NA than group NC,and that of ZO-1 and Occludin proteins in the small intestine wall was significantly higher in group TSA than those in group TS(P<0.05).(4)The group NA exhibited significantly lower LPS concentrations in the gastrocne-mius muscle compared to group NC(P<0.05).Similarly,the group TSA showed reduced LPS levels in both serum and gastrocnemius compared to group TS(P<0.05).The relative protein expression of p-IκBα and p-NF-κB p65 significantly decreased in group TSA compared to group TS(P<0.05).The group TS displayed elevated concentrations of pro-inflammatory cytokines(TNF-α,IL-1β)in serum and gastrocnemius compared to group NC(P<0.01,P<0.05).The group TSA demonstrated significantly lower levels of TNF-α and IL-6 in these tissues in comparison with group TS(P<0.05,P<0.01).Conclusion Aerobic exercise can improve gut microbiota α diversity,adjusting its composition,improv-ing intestinal mucosal barrier function,reducing the LPS-induced pro-inflammatory response,and delay-ing skeletal muscle atrophy.The underlying mechanism may involve down regulation of TLR4/MyD88/NF-κB signaling in skeletal muscle,which has a positive effect on Gut-Muscle Axis.

Objective To investigate the effects of aerobic exercise and different exercise habits on skeletal muscle function and the possible molecular mechanisms in colorectal cancer-loaded mice.Methods Thirty-five 5-week-old BABL/c male mice were acclimatized to feeding for 1 week and then divided randomly into the following groups:control(D),tumor(M),exercise preconditioning(QAM),lifetime exercise(AM),and exercise(HAM)groups(n=7 mice per group).Mice in the QAM and AM groups underwent aerobic exercise regimen 1 from weeks 2～6.At week 7,mice in the experimental groups received 0.2 mL of CT26 colon cancer cell suspension subcutaneously in the dorsal aspect of the left hind limb,while control mice received 0.2 mL of saline at the corresponding site.Mice in the AM and HAM groups were subjected to aerobic exercise regimen 2 for weeks 7～9.The general status and skeletal muscle mass and function were monitored in all mice throughout the experiments.After completion of the experiment,samples were collected and the cross-sectional area of gastrocnemius muscle fibers(CSA)was measured by hematoxylin and eosin staining and expression levels of proteins related to synthesis and catabolism of the gastrocnemius muscle were analyzed by Western Blot.Results(1)The weight ratio of the gastrocnemius muscle was significantly lower in mice in the M,QAM,and HAM groups compared with group D,and was significantly higher in AM mice compared with M,QAM,and HAM mice.(2)Grip strength,endurance,skeletal muscle circumference,and CSA were significantly lower in group D mice compared with the other groups,and was most enhanced in group HAM.Endurance and CSA were consistently enhanced in groups QAM,AM,and HAM.(3)Muscle RING-finger protein-1(MuRF1)expression levels were significantly lower in groups M,QAM,AM,and HAM than in group D,significantly lower in groups AM and HAM than in group M,and significantly lower in group HAM than in group QAM.(4)Fibronectin type Ⅲ domain-containing protein 5(FNDC5)expression levels were significantly lower in group M than in groups D and QAM.(5)Peroxisome proliferator-activated receptor gamma coactivator 1-alpha expression levels were significantly lower in the QAM and HAM groups compared with group M.(6)Expression levels of phospho(p)-AMP-activated protein kinase(AMPK)/AMPK and p-AMPK were significantly higher in group QAM than in groups D and M,p-AMPK expression significantly lower in groups AM and HAM was than in group QAM,and AMPK expression was significantly lower in groups QAM,AM,and HAM than in group D.Conclusions Exercise preconditioning and continuous aerobic exercise can improve skeletal muscle mass and function in CT26 colon cancer-loaded mice by activating AMPK phosphorylation to stimulate skeletal muscle secretion of FNDC5,thereby regulating the expression of MuRF1 protein.

Objective To investigate the effect of aerobic exercise preconditioning on cardiac function in mice bearing subcutaneous transplantable tumors and explore the potential molecular mechanisms.Methods Twenty-four 6-week-old male BALB/c mice were selected.After an acclimation period,they were randomly assigned to a control group(C),a tumor group(M)and an exercise-preconditioning plus tumor group(EM),each of eight.The EM group underwent a 4-week aerobic exercise interven-tion.Meanwhile,the C group received 0.2 ml of physiological saline injected subcutaneously on the dorsum of the proximal left hind limb,while the M and EM groups were inoculated at the same site with 0.2 ml of a CT26.WT colon carcinoma cell suspension.Three weeks after inoculation,cardiac function was assessed by transthoracic echocardiography.Hearts were then harvested for hematoxylin-eo-sin staining to evaluate cardiomyocyte cross-sectional area(CSA),while Western blot was performed to determine the expression of proteins related to myocardial protein synthesis and degradation.Results(1)Compared with group C,group M exhibited significantly lower body weight and heart weight(P<0.05),and reduced left ventricular ejection fraction(LVEF),fractional shortening(FS)and stroke vol-ume(SV)(P<0.05),as well as cardiomyocyte CSA(P<0.01)and total mTOR(P<0.01),phosphory-lated mTOR(p-mTOR)(P<0.01)and the p-mTOR/mTOR ratio(P<0.05),but a significant increase in the expression of the muscle ring finger 1(MuRF-1)and the muscle atrophy F-box(MAFbx/Atrog-in-1)(P<0.01 and P<0.05,respectively).(2)Compared with group M,group EM showed significant-ly greater heart weight(P<0.05);increased left ventricular posterior wall thickness at end-diastole(LVPWd),LVEF,FS and SV(P<0.05),as well as the larger cardiomyocyte CSA and total mTOR,p-mTOR and the p-mTOR/mTOR ratio(P<0.05),but reduced MuRF-1 and Atrogin-1 expression(P<0.05).Conclusion Four weeks of aerobic exercise preconditioning ameliorated myocardial atrophy and improved systolic cardiac function in mice bearing subcutaneously transplanted CT26 tumors.Such beneficial effects may be associated with exercise-induced down regulation of the protein degradation mediators MuRF-1 and Atrogin-1 and up regulation of total mTOR and p-mTOR.

Objective To investigate the effects of oral probiotics on gut microbiota diversity,colony structure,and intergroup differences in mice with subcutaneous colon cancer tumors,based on 16S rRNA sequencing technology.Methods Twenty-four 6-week-old male BALB/c mice were divided randomly into normal control group(NC group,n = 8),model group(M group,n = 8),and probiotic + model group(PM group,n = 8)after adaptive feeding for 1 week.Mice in the PM group were given 200 μL probiotic mixed solution(Bifidobacterium longum and Lactobacillus delbrueckii subsp.bulgaricus mixed lyophilized powder,2×108 colony-forming units)by gavage three times/week for 7 weeks,while the M group and PM group received 200 μL normal saline.At 10 weeks old,0.2 mL CT26.WT cell suspension(1×107/mL)was inoculated subcutaneously into the left hind limbs of M group and PM group,while NC group were inoculated with 0.2 mL normal saline.The general state of mice was observed,the growth of subcutaneous tumor was monitored,and the changes of intestinal flora structure were detected by 16S rRNA sequencing.Results The subcutaneous tumors of the M group were prominent,and the subcutaneous tumor volume and weight of the PM group were significantly reduced(P<0.05).Compared with NC group,Alpha diversity index was lower in the M group,and a significant difference of Beta diversity inter groups(P<0.01).And supplementation of probiotics had a certain effect on gut microbiota diversity in the M group.Compared with M group,the relative abundance of Bacteroidetes,Proteobacteria,Muribaculaceae,Bacteroides were higher in the PM group,while the relative abundance of Firmicutes,Desulfobacterota,Lachnospiraceae_NK4A136_group,Alistipes were lower in the PM group.LEfSe analysis showed that Muribaculaceae and Bacteroides in the PM group were different species with high abundance(LDA values>4).Conclusions Oral probiotics may improve the gut microbiota by increasing the relative abundance of beneficial Muribaculaceae and Bacteroides in subcutaneous tumors in mice with colon cancer.

Objective: To explore the short and mid-term efficacy of device closure of patent foramen ovale (PFO) for treating the patients with PFO combining cryptogenic stroke (CS) and transient ischemic attack (TIA). Methods: A total of 56 PFO patients with CS and TIA receiving device closure in our hospital from 2009-05 to 2015-12 were retrospectively studied. Transthoracic echocardiography (TTE), electrocardiogram (ECG), chest X-ray were examined at 24h, 1 month, 3 and 6 months after theoperation; telephone visit was conducted every 6 months thereafter. Results: There were 54/56 PFO patients combining CS and 2 combining TIA; 53 (94.6%)patients received PFO occluder from Starway medical technology. Aspirin was used for 6 months after the operation. The patients were followed-up for the average of (34.67±23.24) months. No body suffered from post-operative stroke and TIA; no residual shunt was observed. Conclusion: The short and mid-term efficacy of device closure has been satisfactory for treating the patients with PFO combining CS and TIA; its overall clinical value should be further investigated in large population and long-term study.

Objective:To test the validity and reliability of the Chinese version of the Snaith-Hamilton Pleasure Scale (SHAPS).Methods:Totally 166 patients with major depression completed the Chinese version of SHAPS,Beck Depression Inventory (BDI-Ⅰ) and Hamilton Depression Scale (HAMD-17).The validity was tested by using correlation analysis and exploratory factor analysis.The reliability was tested by using internal consistency coefficient and Pearson correlation analysis.Results:The ICCs between three different Scale score were 0.90 and 0.91(Ps ＜ 0.001).Exploratory factor analysis identified 2 common factors,which explained 62.8％ of total variancel.The Cronbach coefficient of SHAPS was 0.94.The test-retest coefficient was 0.91 (P ＜ 0.001).The intercorrelations of 14 items ranged from 0.30 to 0.93 and item-total correlations ranged from 0.61 to 0.79.Conclusion:The Chinese version of SHAPS has good validity and reliability,it could be used to assess anhedonia in Chinese patients with major depression.

Objective To analyze the residents′ willingness to hierarchical medical system and their intention of first visit hospitals.Methods Customized questionnaire was used for in-home survey of 1 500 households in three cities of Shandong province,with 773 of the residents aware of hierarchical medical system used as the study samples.Descriptive analysis and multi factor Logistic regression were employed for statistical analysis of their intention.Results 85.0% the residents expressed willingness to embrace hierarchical medical system.55.0% of the residents prefer primary medical institutions as their first visit in case of mild illness,a choice chosen by more rural residents(58.9%)than urban residents(48.0%),a difference of statistical significance(P<0.01).In the case of severe illness,87.3% of them prefer major hospitals as their first visit,and more urban residents(94.3%)take this option than the rural residents(83.3%),a difference of statistical significance(P<0.01).Service ability of medical institutions and residents′ concept for medical service played a key role in their choice of first visit hospitals.Conclusions The key to ensuring the implementation effect of hierarchical medical system lies in better service ability of primary medical institutions and correction of residents′ irrational medication concepts.