Objective To investigate the effect of exercise during pregnancy on L-type Ca2+ (CaV1.2) channel in mesenteric artery from spontaneously hypertensive rats (SHR) offspring. Methods Female (11 weeks old) and male (12 weeks old) SHR, female (11 weeks old) and male (12 weeks old) WKY rats were selected to use for breeding. The day when the vaginal bolt was found was considered day 1 of gestation. The pregnant rats were randomly divided into four groups: WKY sedentary group (WKY-SED), WKY exercise group (WKY-EX), SHR sedentary group (SHR-SED) and SHR exercise group (SHR-EX). The exercise groups were subjected to swimming at the first day of pregnancy, 1h/d，6 days/week for 3 weeks. The 6-month-old male offspring were operated with femoral arterial and venous cannulation, and the blood pressure after intravenous (i.v.) injection of CaV1.2 channel opener BayK8644 and blocker nifedipine were monitored in vivo. In vitro study, the mesenteric arteries were removed and used for isometric contraction studies. The non-selective NOS inhibitor Nω-nitro-L-arginine methyl ester (L-NAME, 100 μM) was added after 60mM KCl measurement. To investigate the contribution of CaV1.2 channels in vascular tone regulation, the vascular responses to nifedipine (10−9–10−5M) were examined. Western blot was applied to examine the protein expression levels of CaV1.2 channel. Results (1) The mean arterial pressure(MAP) were higher in the 6M offspring of SHR-SED group than that of WKY-SED group (P＜0.01), but there was no significant difference between the 6M offspring of SED and Ex groups. (2) The pressor responses induced by i.v. injection of BayK8644（0.1mg/Kg）were increased in the 6M offspring of SHR-SED group (P＜0.05) compared with the WKY-SED group. Exercise during pregnancy markedly decreased the pressor responses in 6M offspring of SHR-EX group (P＜0.05). (3) Compared with the 6M offspring of WKY-SED group, the depressor responses induced by i.v. injection of nifedipine（1mg/Kg）were increased in the 6M offspring of SHR-SED group (P＜0.01). Exercise during pregnancy markedly attenuated the depressor responses in 6M offspring of SHR-EX group (P＜0.05). (4) The isometric contraction study revealed that nifedipine induced concentration-dependent vasorelaxation in mesenteric artery precontracted with noradrenaline. The sensitivity of tissues to nifedipine in 6M offspring of SHR-SED group was significantly higher than that of WKY-SED group (P＜0.01). Exercise during pregnancy normalized the increased sensitivity of tissues to nifedipine in 6M offspring of SHR (P＜0.05). (5) Compared with the 6M offspring of WKY-SED group, the protein expression of CaV1.2α1C was significantly increased in SHR-SED group(P＜0.01). Exercise during pregnancy markedly inhibited the expression of CaV1.2α1C in 6M offspring of SHR-EX group (P＜0.05). Conclusions Pregnancy exercise has no significant effect on basic blood pressure in 6M offspring of SHR; but the increased function and protein expression of CaV1.2 c
{"title":"PO-139 Effects of Exercise During Pregnancy on CaV1.2 Channel in Mesenteric Artery from SHR Offspring","authors":"Shanshan Li, F. Zeng, Lijun Shi","doi":"10.14428/EBR.V1I4.10423","DOIUrl":"https://doi.org/10.14428/EBR.V1I4.10423","url":null,"abstract":"Objective To investigate the effect of exercise during pregnancy on L-type Ca2+ (CaV1.2) channel in mesenteric artery from spontaneously hypertensive rats (SHR) offspring. \u0000Methods Female (11 weeks old) and male (12 weeks old) SHR, female (11 weeks old) and male (12 weeks old) WKY rats were selected to use for breeding. The day when the vaginal bolt was found was considered day 1 of gestation. The pregnant rats were randomly divided into four groups: WKY sedentary group (WKY-SED), WKY exercise group (WKY-EX), SHR sedentary group (SHR-SED) and SHR exercise group (SHR-EX). The exercise groups were subjected to swimming at the first day of pregnancy, 1h/d，6 days/week for 3 weeks. The 6-month-old male offspring were operated with femoral arterial and venous cannulation, and the blood pressure after intravenous (i.v.) injection of CaV1.2 channel opener BayK8644 and blocker nifedipine were monitored in vivo. In vitro study, the mesenteric arteries were removed and used for isometric contraction studies. The non-selective NOS inhibitor Nω-nitro-L-arginine methyl ester (L-NAME, 100 μM) was added after 60mM KCl measurement. To investigate the contribution of CaV1.2 channels in vascular tone regulation, the vascular responses to nifedipine (10−9–10−5M) were examined. Western blot was applied to examine the protein expression levels of CaV1.2 channel. \u0000Results (1) The mean arterial pressure(MAP) were higher in the 6M offspring of SHR-SED group than that of WKY-SED group (P＜0.01), but there was no significant difference between the 6M offspring of SED and Ex groups. (2) The pressor responses induced by i.v. injection of BayK8644（0.1mg/Kg）were increased in the 6M offspring of SHR-SED group (P＜0.05) compared with the WKY-SED group. Exercise during pregnancy markedly decreased the pressor responses in 6M offspring of SHR-EX group (P＜0.05). (3) Compared with the 6M offspring of WKY-SED group, the depressor responses induced by i.v. injection of nifedipine（1mg/Kg）were increased in the 6M offspring of SHR-SED group (P＜0.01). Exercise during pregnancy markedly attenuated the depressor responses in 6M offspring of SHR-EX group (P＜0.05). (4) The isometric contraction study revealed that nifedipine induced concentration-dependent vasorelaxation in mesenteric artery precontracted with noradrenaline. The sensitivity of tissues to nifedipine in 6M offspring of SHR-SED group was significantly higher than that of WKY-SED group (P＜0.01). Exercise during pregnancy normalized the increased sensitivity of tissues to nifedipine in 6M offspring of SHR (P＜0.05). (5) Compared with the 6M offspring of WKY-SED group, the protein expression of CaV1.2α1C was significantly increased in SHR-SED group(P＜0.01). Exercise during pregnancy markedly inhibited the expression of CaV1.2α1C in 6M offspring of SHR-EX group (P＜0.05). \u0000Conclusions Pregnancy exercise has no significant effect on basic blood pressure in 6M offspring of SHR; but the increased function and protein expression of CaV1.2 c","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"19 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81807545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chuanjun Wang, Jianmin Cao, Xingang Che, Bingxin Qiu
Objective Since the Mexican Olympic Games, altitude training has attracted the attention of international sports science circle with its remarkable training effect, which is regarded as one of the effective methods to improve aerobic capacity. With the improvement of altitude training by scholars at home and abroad, hypoxic training has gradually entered the public view. Hypoxic training aims to achieve hypoxic stimulation by artificially simulating the natural hypoxic environment in the plateau or simulating the biological effects of hypoxic on human body. However, whether the aerobic capacity can be improved through hypoxic training requires us to further study and explore the mechanism of hypoxic training. This study explored the mechanism of hypoxic exercise training by stimulating long-term hypoxic exercise training for overweight or obese adult males. Methods In this study, 40 male (aged 18—47 years) overweight/obese subjects were recruited. No physical condition was abnormal after physical examination, and BMI ≥ 24 was overweight, while BMI ≥ 28 was obese. All subjects were paired according to their weight and divided into the hypoxic group and the normoxic group, the exercise intervention lasted for 6 weeks. The exercise intervention program consists of 30min strength training and 30min endurance training, with 5 minutes of warm-up and finishing activities before and after training. Strength training uses dumbbells, chooses 12RM weight, exercise with 8 actions, which are dead lift, upright row, squat, shoulder press, calf Jump, advance junge, biceps curl and triceps extension, each action 2 Group, rest between groups for 30s. Endurance training grade 0°treadmill, speed range according to the target heart rate adjustment, the target heart rate interval computation method for 60% HRmax—70% HRmax. Among them, subjects in the hypoxic group wore inhaled low-oxygen devices, which enabled them to exercise in a hypoxic environment. The oxygen content of the inhaled mixed gas was 16%. The subjects in the aerobic group exercised in an aerobic environment. Nutritional education was administered to all subjects prior to the start of the exercise intervention, but diet was not restricted during the intervention. Fasting venous blood before and after intervention, the detection of hemoglobin (Hb) and erythropoietin (EPO), hypoxia-inducible factor 1 alpha (HIF1α), vascular endothelial growth factor (VEGF) and testosterone (T). All test results are the mean ± standard deviation, data comparison between groups using nonparametric the Mann-Whitney U test, data comparison in the group using nonparametric Wilcoxon match the symbol rank test, the significance level of P<0.05, very significance level of P<0.01. Results (1) After 6 weeks of intervention, Hb levels were elevated in the hypoxic group, but there was no statistically difference compared with the pre-intervention (P>0.05). And the change rate of Hb in the hypoxic group was higher than that in the
{"title":"PO-070 Effects of hypoxic exercise training on aerobic capacity-related proteins in overweight/obese adult males","authors":"Chuanjun Wang, Jianmin Cao, Xingang Che, Bingxin Qiu","doi":"10.14428/EBR.V1I3.11283","DOIUrl":"https://doi.org/10.14428/EBR.V1I3.11283","url":null,"abstract":"Objective Since the Mexican Olympic Games, altitude training has attracted the attention of international sports science circle with its remarkable training effect, which is regarded as one of the effective methods to improve aerobic capacity. With the improvement of altitude training by scholars at home and abroad, hypoxic training has gradually entered the public view. Hypoxic training aims to achieve hypoxic stimulation by artificially simulating the natural hypoxic environment in the plateau or simulating the biological effects of hypoxic on human body. However, whether the aerobic capacity can be improved through hypoxic training requires us to further study and explore the mechanism of hypoxic training. This study explored the mechanism of hypoxic exercise training by stimulating long-term hypoxic exercise training for overweight or obese adult males. \u0000Methods In this study, 40 male (aged 18—47 years) overweight/obese subjects were recruited. No physical condition was abnormal after physical examination, and BMI ≥ 24 was overweight, while BMI ≥ 28 was obese. All subjects were paired according to their weight and divided into the hypoxic group and the normoxic group, the exercise intervention lasted for 6 weeks. The exercise intervention program consists of 30min strength training and 30min endurance training, with 5 minutes of warm-up and finishing activities before and after training. Strength training uses dumbbells, chooses 12RM weight, exercise with 8 actions, which are dead lift, upright row, squat, shoulder press, calf Jump, advance junge, biceps curl and triceps extension, each action 2 Group, rest between groups for 30s. Endurance training grade 0°treadmill, speed range according to the target heart rate adjustment, the target heart rate interval computation method for 60% HRmax—70% HRmax. Among them, subjects in the hypoxic group wore inhaled low-oxygen devices, which enabled them to exercise in a hypoxic environment. The oxygen content of the inhaled mixed gas was 16%. The subjects in the aerobic group exercised in an aerobic environment. Nutritional education was administered to all subjects prior to the start of the exercise intervention, but diet was not restricted during the intervention. Fasting venous blood before and after intervention, the detection of hemoglobin (Hb) and erythropoietin (EPO), hypoxia-inducible factor 1 alpha (HIF1α), vascular endothelial growth factor (VEGF) and testosterone (T). All test results are the mean ± standard deviation, data comparison between groups using nonparametric the Mann-Whitney U test, data comparison in the group using nonparametric Wilcoxon match the symbol rank test, the significance level of P<0.05, very significance level of P<0.01. \u0000Results (1) After 6 weeks of intervention, Hb levels were elevated in the hypoxic group, but there was no statistically difference compared with the pre-intervention (P>0.05). And the change rate of Hb in the hypoxic group was higher than that in the","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"4 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80315742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective This study was designed to investigate the effects of curcumin on inflammatory factors and ECM expression in exercise-induced renal ischemia reperfusion injury in rats. Methods Sixty 7-week-old male SD rats were divided randomly into three groups: group A (normal control group, n=12), group B (overtraining group, n=24) and group C (curcumin + overtraining group, n=24). Group B and C performed 6 weeks of incremental load training on the treadmill. 24 hours after the last training, the rats were anesthetized intraperitoneally, the morphology of renal tissue and the deposition of glomerular ECM were observed using light microscope，the related biochemical indexes were tested. Results (1) the renal structure of rats in group A were normal, histopathological changes were observed in group B and C, Paller score of group B were significantly higher than group A(P <0.01), and that of group C were significantly lower than group B(P <0.05). (2) Blood urea nitrogen (BUN) and serum creatinine (Scr), serum and renal inflammatory factors , TGF-β protein expression level and glomerular ECM deposition of group B were significantly higher than group A(P<0.01) and those of group C were lower than group B(P<0.05). Conclusions Supplementation of curcumin can effectively protect rats from exercise-induced renal ischemia reperfusion injury, by inhibiting the up-regulation of inflammatory cytokines and TGF-β expression and maintaining the dynamic balance of ECM .
{"title":"PO-032 The protective effects of curcumin on exercise-induced renal ischemia reperfusion injury in rats","authors":"Fang-yuan Li, Jianmin Cao, Hai-tao Zhou, Y. Niu","doi":"10.14428/EBR.V1I3.9973","DOIUrl":"https://doi.org/10.14428/EBR.V1I3.9973","url":null,"abstract":"Objective This study was designed to investigate the effects of curcumin on inflammatory factors and ECM expression in exercise-induced renal ischemia reperfusion injury in rats. \u0000Methods Sixty 7-week-old male SD rats were divided randomly into three groups: group A (normal control group, n=12), group B (overtraining group, n=24) and group C (curcumin + overtraining group, n=24). Group B and C performed 6 weeks of incremental load training on the treadmill. 24 hours after the last training, the rats were anesthetized intraperitoneally, the morphology of renal tissue and the deposition of glomerular ECM were observed using light microscope，the related biochemical indexes were tested. \u0000Results (1) the renal structure of rats in group A were normal, histopathological changes were observed in group B and C, Paller score of group B were significantly higher than group A(P <0.01), and that of group C were significantly lower than group B(P <0.05). \u0000(2) Blood urea nitrogen (BUN) and serum creatinine (Scr), serum and renal inflammatory factors , TGF-β protein expression level and glomerular ECM deposition of group B were significantly higher than group A(P<0.01) and those of group C were lower than group B(P<0.05). \u0000Conclusions Supplementation of curcumin can effectively protect rats from exercise-induced renal ischemia reperfusion injury, by inhibiting the up-regulation of inflammatory cytokines and TGF-β expression and maintaining the dynamic balance of ECM .","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"246 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83242869","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective The aim of this meta-analysis was to assess the function index change trend of some fighting event-group athletes during weight reduction before competition. Methods Pub Med, MEDLINE, EBSCO, CNKI, Wanfang and VIP were searched to collect the scientific literatures about the change trend of body composition and function index of wrestling, judo and taekwondo athletes during weight reduction before the competition. The quality of literature was critically appraised, and data were extracted by 2 reviewers independently. Meta-analysis was conducted for eligible literatures. Results A total of 14 studies were included in this meta-analysis. Stata software was used for data processing to observe the trend of body composition and function index during the process of weight reduction before competition. The average weight of athletes in judo，wrestling and taekwondo decreased by 5.23kg and the body fat percent decreased by 2.13% on average. Testosterone decreased by 3.93nmol/L on average and cortisol increased by 68.55noml/L on average. Hemoglobin decreased by 1.84g/L on average and blood urea decreased by 0.026 mmol/L on average. It was also found that the body weight and the body fat percent decreased significantly during the fast control period (P < 0.05), the decrease of the body fat percent during the fast control period was significantly higher than that in the slow control period. Testosterone decreased significantly (P < 0.01) during the slow control period and continued downward trend until pre-competition. Hemoglobin in the fast control period and pre-competition significantly lower than that before weight control (P < 0.05). The level of blood urea in fast control period was significantly higher than that in slow control period (P < 0.01). Conclusions The decrease of body weight and body fat percent was concentrated in fast control period to pre-competition stag. Testosterone levels began to decline at the slow control period and lasted until the pre-competition.It is inconsistent with the expected results of coaches and athletes.We need further search for reasons.
{"title":"PO-270 Function Index Change Trend Of Fighting Events-Group Athletes During Weight Reduction Before Competition: A Meta-Analysis","authors":"Fanyang Zhou","doi":"10.14428/EBR.V1I5.11193","DOIUrl":"https://doi.org/10.14428/EBR.V1I5.11193","url":null,"abstract":"Objective The aim of this meta-analysis was to assess the function index change trend of some fighting event-group athletes during weight reduction before competition. \u0000Methods Pub Med, MEDLINE, EBSCO, CNKI, Wanfang and VIP were searched to collect the scientific literatures about the change trend of body composition and function index of wrestling, judo and taekwondo athletes during weight reduction before the competition. The quality of literature was critically appraised, and data were extracted by 2 reviewers independently. Meta-analysis was conducted for eligible literatures. \u0000Results A total of 14 studies were included in this meta-analysis. Stata software was used for data processing to observe the trend of body composition and function index during the process of weight reduction before competition. The average weight of athletes in judo，wrestling and taekwondo decreased by 5.23kg and the body fat percent decreased by 2.13% on average. Testosterone decreased by 3.93nmol/L on average and cortisol increased by 68.55noml/L on average. Hemoglobin decreased by 1.84g/L on average and blood urea decreased by 0.026 mmol/L on average. It was also found that the body weight and the body fat percent decreased significantly during the fast control period (P < 0.05), the decrease of the body fat percent during the fast control period was significantly higher than that in the slow control period. Testosterone decreased significantly (P < 0.01) during the slow control period and continued downward trend until pre-competition. Hemoglobin in the fast control period and pre-competition significantly lower than that before weight control (P < 0.05). The level of blood urea in fast control period was significantly higher than that in slow control period (P < 0.01). \u0000Conclusions The decrease of body weight and body fat percent was concentrated in fast control period to pre-competition stag. Testosterone levels began to decline at the slow control period and lasted until the pre-competition.It is inconsistent with the expected results of coaches and athletes.We need further search for reasons. \u0000 ","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"14 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77756214","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective The paper provides an overview of probiotics on the ergogenic effect,which is prepared to demonstrate that moderate doses of probiotics have many benefits for the physical exercise，the development of related probiotic functional food is of great benefit to the majority of sports people. Methods while probiotics as one of the most popular nutritional supplements,Many studies have shown that probiotics mixture(mainly lactic acid bacteria and bifidobacterium) are supplemented can improve intestinal microflora, reduce intestinal permeability, control endotoxin transport and reduce the incidence of intestinal leakage;Promoteing the metabolism of short-chain fatty acids (SCAFs), provide metabolic energy, enhance muscle strength and endurance, optimize muscle fiber composition, improve anti-fatigue ability;Improving the ability of immune regulation, such as reducing upper respiratory tract and gastrointestinal tract infection symptoms ;Reducing the oxidative stress level, increasing the scavenging rate of superoxide anion radical, reducing capacity and iron ion chelating activity can alleviate the oxidative stress after heavy exercise；Supplementation of probiotics can reconstruct intestinal flora, and intestinal flora can affect bone health and enhance calcium absorption；ther studies have confirmed that probiotics can regulate emotional disorders through the hypothalamus-pituitary-adrenal axis and relieve anxiety and depression, which also contributes to the mental health of the exercise population Results In addition, the amount of exercise, the variety of probiotics, the quantity of supplementation and the time of supply in the reference literature are different.Therefore, the experimental results are different, and the mechanism remains to be further studied. Conclusions Moderate doses of probiotics have many benefits for the physical exercise，the development of related probiotic functional food is of great benefit to the majority of sports people.
{"title":"PO-269 An overview of probiotics on the ergogenic effect","authors":"Yali Xia","doi":"10.14428/EBR.V1I5.11183","DOIUrl":"https://doi.org/10.14428/EBR.V1I5.11183","url":null,"abstract":"Objective The paper provides an overview of probiotics on the ergogenic effect,which is prepared to demonstrate that moderate doses of probiotics have many benefits for the physical exercise，the development of related probiotic functional food is of great benefit to the majority of sports people. \u0000Methods while probiotics as one of the most popular nutritional supplements,Many studies have shown that probiotics mixture(mainly lactic acid bacteria and bifidobacterium) are supplemented can improve intestinal microflora, reduce intestinal permeability, control endotoxin transport and reduce the incidence of intestinal leakage;Promoteing the metabolism of short-chain fatty acids (SCAFs), provide metabolic energy, enhance muscle strength and endurance, optimize muscle fiber composition, improve anti-fatigue ability;Improving the ability of immune regulation, such as reducing upper respiratory tract and gastrointestinal tract infection symptoms ;Reducing the oxidative stress level, increasing the scavenging rate of superoxide anion radical, reducing capacity and iron ion chelating activity can alleviate the oxidative stress after heavy exercise；Supplementation of probiotics can reconstruct intestinal flora, and intestinal flora can affect bone health and enhance calcium absorption；ther studies have confirmed that probiotics can regulate emotional disorders through the hypothalamus-pituitary-adrenal axis and relieve anxiety and depression, which also contributes to the mental health of the exercise population \u0000Results In addition, the amount of exercise, the variety of probiotics, the quantity of supplementation and the time of supply in the reference literature are different.Therefore, the experimental results are different, and the mechanism remains to be further studied. \u0000Conclusions Moderate doses of probiotics have many benefits for the physical exercise，the development of related probiotic functional food is of great benefit to the majority of sports people. \u0000 ","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"101 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81420258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective In order to elucidate the target proteins of exercise-induced stress, differential expression of proteins in urine from male athletes after 30 km running was evaluated by proteomics. Methods Urine samples were collected from ten male runners before and after 30 km running. The differential expression profile of urine proteins was investigated by using a proteomic approach based on two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) procedures. Results Totally 1011 ± 243 and 1737 ± 15 protein spots were detected from 2-DE of urine samples harvested from the subjects before and after 30 km running. In addition, 110 protein spots with differential expression were achieved. After exercise, 10 proteins with ≥5 fold up-regulation and 23 new proteins were observed; 18 protein spots with ≥5 fold down-regulation were observed and 6 proteins were disappeared. The target proteins were identified as zinc-α2-glycoprotein, albumin, vitamin D-binding protein, prostate-specific antigen, β-actin and Bence-Jones protein (BJP), which are correlated with the change of energy metabolism pathway, material transport and stress protection in the body. Conclusions An obviously differential protein expression profile in urine samples from the subjects after 30 km running is achieved, which provides a new idea for further exploring exercise-induced proteinuria. However, further studies need to develop urine protein profiles of athletes, which will benefit for monitoring sport competition and doping control as well as excessive exercise-induced diseases.
{"title":"PO-061 Differential expression of proteins in urea from male athletes after 30 km running evaluated by proteomics","authors":"Wen-tao Lin, Haiqi Lin","doi":"10.14428/EBR.V1I3.10813","DOIUrl":"https://doi.org/10.14428/EBR.V1I3.10813","url":null,"abstract":"Objective In order to elucidate the target proteins of exercise-induced stress, differential expression of proteins in urine from male athletes after 30 km running was evaluated by proteomics. \u0000Methods Urine samples were collected from ten male runners before and after 30 km running. The differential expression profile of urine proteins was investigated by using a proteomic approach based on two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) procedures. \u0000Results Totally 1011 ± 243 and 1737 ± 15 protein spots were detected from 2-DE of urine samples harvested from the subjects before and after 30 km running. In addition, 110 protein spots with differential expression were achieved. After exercise, 10 proteins with ≥5 fold up-regulation and 23 new proteins were observed; 18 protein spots with ≥5 fold down-regulation were observed and 6 proteins were disappeared. The target proteins were identified as zinc-α2-glycoprotein, albumin, vitamin D-binding protein, prostate-specific antigen, β-actin and Bence-Jones protein (BJP), which are correlated with the change of energy metabolism pathway, material transport and stress protection in the body. \u0000Conclusions An obviously differential protein expression profile in urine samples from the subjects after 30 km running is achieved, which provides a new idea for further exploring exercise-induced proteinuria. However, further studies need to develop urine protein profiles of athletes, which will benefit for monitoring sport competition and doping control as well as excessive exercise-induced diseases.","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"41 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80852175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective The purpose of the study was to explore the mechanism underlying physical training in cool environment (20˚C) improving tolerance to exercise-heat stress. Methods Rats were randomly divided into aerobic exercise group(AE group) and control group (C group). Rats of AE group undertook 5 weeks aerobic physical training in cool environment, rats of C group were kept sedentary. Through Western blotting method, the expression of HSP70 in rats motor cortex of both groups was detected. The hydroxyl radical scavenging capacity (HRSC), malondialdehyde (MDA), superoxide dismutase (SOD) activity, xanthine oxidase (XOD) and glutathion peroxidase (GPX) in rat motor cortex were detected immediately after 1h heat exposure or exhaustive exercise in heat (38-40 ˚C). The exhaustion time and changes of rectal temperature during exhaustive exercise were recorded. Results Results showed that, the HSP70 expression of AE group is higher than C group (P<0.05). After 1h heat exposure or exhaustive exercise, the XOD and MDA of AE group are all lower than C group (P<0.05). The HRSC, SOD and GSH-PX of AE group are all higher than C group (P<0.05). At 30, 45 and 60 min time point, the rectal temperature of AE group are all lower than C group (P<0.05). The exhaustion time of AE group is higher than C group(P<0.05). Conclusions Our results suggest that the enhanced expression of HSP70 and the decreased rise rate of core temperature, may result in the lower oxidative damage in rat brain, this may be one important factor for physical training in cool environment improving tolerance to exercise-heat stress.
{"title":"PO-251 Aerobic physical training in cool environment protects rat brain against oxidative damage during rest and exercise in heat through the increased expression level of HSP70","authors":"Dalei Wang","doi":"10.14428/EBR.V1I5.10993","DOIUrl":"https://doi.org/10.14428/EBR.V1I5.10993","url":null,"abstract":"Objective The purpose of the study was to explore the mechanism underlying physical training in cool environment (20˚C) improving tolerance to exercise-heat stress. \u0000Methods Rats were randomly divided into aerobic exercise group(AE group) and control group (C group). Rats of AE group undertook 5 weeks aerobic physical training in cool environment, rats of C group were kept sedentary. Through Western blotting method, the expression of HSP70 in rats motor cortex of both groups was detected. The hydroxyl radical scavenging capacity (HRSC), malondialdehyde (MDA), superoxide dismutase (SOD) activity, xanthine oxidase (XOD) and glutathion peroxidase (GPX) in rat motor cortex were detected immediately after 1h heat exposure or exhaustive exercise in heat (38-40 ˚C). The exhaustion time and changes of rectal temperature during exhaustive exercise were recorded. \u0000Results Results showed that, the HSP70 expression of AE group is higher than C group (P<0.05). After 1h heat exposure or exhaustive exercise, the XOD and MDA of AE group are all lower than C group (P<0.05). The HRSC, SOD and GSH-PX of AE group are all higher than C group (P<0.05). At 30, 45 and 60 min time point, the rectal temperature of AE group are all lower than C group (P<0.05). The exhaustion time of AE group is higher than C group(P<0.05). \u0000Conclusions Our results suggest that the enhanced expression of HSP70 and the decreased rise rate of core temperature, may result in the lower oxidative damage in rat brain, this may be one important factor for physical training in cool environment improving tolerance to exercise-heat stress. \u0000 ","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"153 8 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83176557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective Objective:Cortical parvalbumin-expressing inhibitory neurons(PV) control the activity of excitatory neurons and regulate their spike output. The present experiment is to determine the role of PV neuron in the reglution of excitability of primary motor cortex (M1) during the exercise-induced fatigue and possible molecular mechanism. Methods Methods: Male Wistar rats randomly divided into control group(C),exhaustive exercise group(E) and repeated exhaustive exercise group(RE). The gradually increasing load treadmill exercise-induced fatigue model was employed in the Group E and RE.The in vivo multi-channel recording methods was used for recording the neuronal electrophysiological activities of primary motor cortex.To observe the neuron firing rate changes during the rest state,immediately after exhausted exercise and after repeated exhaustive exercise.We also detected the expression of PV positive neurons in the primary motor cortex by the immunofluorescence method. The western blot method was used to determine the expression of calmodulin-dependent protein kinase II (CaMKII)、phosphorylated calmodulin-dependent protein kinase II( pCaMKII) and extracellular signal regulated kinase (ERK) in the primary motor cortex. Results Results：The electrophysioligical results indicated that the neuron firing rate after repeated exhausted excise the neuron firing rate significantly decreased compared with the rest state (P<0.05),but have no significantly changes as compared with exhausted excise;The expression of PV positive neurons in the group of E and RE significantly increased compared with the group C(P<0.01);The western blot results indicated that the protein expression of ERK in group REsignificantly decreased compared with group C, the pCaMKII expression of group RE decreased,but have no statistical difference. Conclusions Conclusion: After exercise-indued fatigue ,the increase of PV positive neuron maybe one reason for the excitability changes in primary motor cortex.the alteraions in the electrical signal may be participate in the regluation of exercise-induced fatigue. pCaMKII and ERK signal pathway may invloved in the molecular mechanism of exercise-induced fatigue.
目的目的:皮层小蛋白表达抑制神经元(PV)控制兴奋性神经元的活动并调节其峰输出。本实验旨在探讨PV神经元在运动性疲劳过程中对初级运动皮层(M1)兴奋性的调控作用及其可能的分子机制。方法:雄性Wistar大鼠随机分为对照组(C)、力竭运动组(E)和重复力竭运动组(RE)。E组和re组采用负荷逐渐增加的跑步机运动性疲劳模型，采用体内多通道记录法记录初级运动皮层神经元电生理活动。观察静息状态下、疲劳运动后及反复疲劳运动后神经元放电速率的变化。免疫荧光法检测原发性运动皮层PV阳性神经元的表达。western blot法检测原代运动皮层钙调素依赖性蛋白激酶II(CaMKII)、磷酸化钙调素依赖性蛋白激酶II(pCaMKII)和细胞外信号调节激酶(ERK)的表达。结果结果:电生理结果显示，重复竭电切除后神经元放电率与静息状态相比显著降低(P<0.05)，但与竭电切除相比无显著变化;E组和RE组PV阳性神经元表达较C组显著升高(P<0.01); western blot结果显示，RE组ERK蛋白表达较C组显著降低C组，RE组pCaMKII表达降低，但差异无统计学意义。结论:运动性疲劳后，PV阳性神经元的增加可能是原代运动皮层兴奋性改变的原因之一。电信号的改变可能参与了运动性疲劳的调节。pCaMKII和ERK信号通路可能参与运动性疲劳的分子机制。
{"title":"OR-034 The parvalbumin positive neuron involved the regulation of motor cortex excitability in the exercise-induced fatigue","authors":"Shuqiang Cui, D. Qiao, Xiaoli Liu","doi":"10.14428/EBR.V1I2.9623","DOIUrl":"https://doi.org/10.14428/EBR.V1I2.9623","url":null,"abstract":"Objective Objective:Cortical parvalbumin-expressing inhibitory neurons(PV) control the activity of excitatory neurons and regulate their spike output. The present experiment is to determine the role of PV neuron in the reglution of excitability of primary motor cortex (M1) during the exercise-induced fatigue and possible molecular mechanism. \u0000Methods Methods: Male Wistar rats randomly divided into control group(C),exhaustive exercise group(E) and repeated exhaustive exercise group(RE). The gradually increasing load treadmill exercise-induced fatigue model was employed in the Group E and RE.The in vivo multi-channel recording methods was used for recording the neuronal electrophysiological activities of primary motor cortex.To observe the neuron firing rate changes during the rest state,immediately after exhausted exercise and after repeated exhaustive exercise.We also detected the expression of PV positive neurons in the primary motor cortex by the immunofluorescence method. The western blot method was used to determine the expression of calmodulin-dependent protein kinase II (CaMKII)、phosphorylated calmodulin-dependent protein kinase II( pCaMKII) and extracellular signal regulated kinase (ERK) in the primary motor cortex. \u0000Results Results：The electrophysioligical results indicated that the neuron firing rate after repeated exhausted excise the neuron firing rate significantly decreased compared with the rest state (P<0.05),but have no significantly changes as compared with exhausted excise;The expression of PV positive neurons in the group of E and RE significantly increased compared with the group C(P<0.01);The western blot results indicated that the protein expression of ERK in group REsignificantly decreased compared with group C, the pCaMKII expression of group RE decreased,but have no statistical difference. \u0000Conclusions Conclusion: After exercise-indued fatigue ,the increase of PV positive neuron maybe one reason for the excitability changes in primary motor cortex.the alteraions in the electrical signal may be participate in the regluation of exercise-induced fatigue. pCaMKII and ERK signal pathway may invloved in the molecular mechanism of exercise-induced fatigue.","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"15 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78730926","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Die Wu, Wei Cao, Beibei Luo, D. Xiang, Peijie Chen
Objective Intestinal epithelial cells are positioned between an anaerobic lumen and a highly metabolic lamina propria, affected by reduced blood flow and tissue hypoxia. Exercise induces blood flow redistribution, leading to hypoperfusion and gastrointestinal (GI) compromise. The hypoxia-inducible factor (HIF) 1α is pivotal in the transcriptional response to oxygen flux. In this study, we hypothesized that exercise induces GI system hypoxia and accumulates HIF-1α. Methods (1) ROSA26 ODD-Luc/+ mouse model (ODD-Luc) was used to detect HIF-1α expression in the intestine (female, 8-week, n=6/group). ODD-Luc mice were randomized into 4 groups: stayed in 21% O2 as the normoxic control (C), exercise (E), injected HIF-1α inhibitor PX-478 before swimming (PS), placed in the chamber containing 9% O2 for 4 hours as the positive control (PC). (2) Exercise models were conducted by volume: Moderate Exercise (ME): mice voluntarily swam for 30 min; Heavy-intensity Exercise (HE): mice swam for 1.5 hours with 5% body weight loads attached to their tails; Long-time Exercise (LE): mice voluntarily swam for 3 hours or till fatigue. Results (1) Exercise increased HIF-1α in the abdominal area. The luciferase activities boosted after exercise, compared to the controls (ME v.s. C, P<0.05; HE v.s. C, P<0.05; LE v.s. C, P<0.05) but no differences among three exercise groups (ME v.s. HE, P>0.99; ME v.s. LE, P>0.99; HE v.s. LE, P>0.99); (2) Exercise altered HIF-1α distribution in the small intestine in a time-dependent manner. The expression of HIF-1α was significantly increased after exercise and gradually reduced to the rest level. The photons increased at the 0th hour after exercise compared to that of the normoxic control (P<0.01). The level of photons then reduced over time, while the 2nd, 4th and 6th hour post-exercise were still greater than that of the normoxic control (2nd hour v.s. C, P<0.01; 4th hour v.s. C, P<0.01; 6th hour v.s. C, P<0.05), and returned to normal after 24 hours (24th hour v.s. C, P>0.99). Conclusions Exercise induced the distribution of HIF-1α in the small intestine. The expression of HIF-1α is shown in a time-dependent manner after exercise.
肠上皮细胞位于厌氧腔和高代谢固有层之间，受血流量减少和组织缺氧的影响。运动诱导血流重新分配，导致灌注不足和胃肠道(GI)受损。缺氧诱导因子(HIF) 1α在对氧通量的转录反应中起关键作用。在本研究中，我们假设运动诱导胃肠道系统缺氧并积累HIF-1α。方法(1)采用ROSA26 ODD-Luc/+小鼠模型(ODD-Luc)检测HIF-1α在肠道中的表达(雌性，8周，n=6/组)。将ODD-Luc小鼠随机分为4组:常氧对照组(C组)、运动组(E组)、游泳前注射HIF-1α抑制剂PX-478组(PS组)、阳性对照组(PC组)。(2)按体积建立运动模型:适度运动(ME):小鼠自愿游泳30 min;高强度运动(HE):小鼠在尾巴附着5%体重负荷的情况下游泳1.5小时;长时间运动(LE):小鼠自愿游泳3小时或至疲劳。结果(1)运动增加腹部HIF-1α。与对照组相比，运动后荧光素酶活性增强(ME vs .s. C, P0.99;ME vs .s. LE, P < 0.99;HE vs .s. LE, P [0.99];(2)运动改变了HIF-1α在小肠中的分布，并呈时间依赖性。HIF-1α的表达在运动后显著升高，并逐渐降低至静止水平。与常氧对照组相比，运动后第0小时光子增加(P0.99)。结论运动可诱导HIF-1α在小肠中的分布。运动后HIF-1α的表达呈时间依赖性。
{"title":"OR-026 Exercise induces HIF-1α redistribution in the small intestine","authors":"Die Wu, Wei Cao, Beibei Luo, D. Xiang, Peijie Chen","doi":"10.14428/EBR.V1I2.9273","DOIUrl":"https://doi.org/10.14428/EBR.V1I2.9273","url":null,"abstract":"Objective Intestinal epithelial cells are positioned between an anaerobic lumen and a highly metabolic lamina propria, affected by reduced blood flow and tissue hypoxia. Exercise induces blood flow redistribution, leading to hypoperfusion and gastrointestinal (GI) compromise. The hypoxia-inducible factor (HIF) 1α is pivotal in the transcriptional response to oxygen flux. In this study, we hypothesized that exercise induces GI system hypoxia and accumulates HIF-1α. \u0000Methods (1) ROSA26 ODD-Luc/+ mouse model (ODD-Luc) was used to detect HIF-1α expression in the intestine (female, 8-week, n=6/group). ODD-Luc mice were randomized into 4 groups: stayed in 21% O2 as the normoxic control (C), exercise (E), injected HIF-1α inhibitor PX-478 before swimming (PS), placed in the chamber containing 9% O2 for 4 hours as the positive control (PC). (2) Exercise models were conducted by volume: Moderate Exercise (ME): mice voluntarily swam for 30 min; Heavy-intensity Exercise (HE): mice swam for 1.5 hours with 5% body weight loads attached to their tails; Long-time Exercise (LE): mice voluntarily swam for 3 hours or till fatigue. \u0000Results (1) Exercise increased HIF-1α in the abdominal area. The luciferase activities boosted after exercise, compared to the controls (ME v.s. C, P<0.05; HE v.s. C, P<0.05; LE v.s. C, P<0.05) but no differences among three exercise groups (ME v.s. HE, P>0.99; ME v.s. LE, P>0.99; HE v.s. LE, P>0.99); (2) Exercise altered HIF-1α distribution in the small intestine in a time-dependent manner. The expression of HIF-1α was significantly increased after exercise and gradually reduced to the rest level. The photons increased at the 0th hour after exercise compared to that of the normoxic control (P<0.01). The level of photons then reduced over time, while the 2nd, 4th and 6th hour post-exercise were still greater than that of the normoxic control (2nd hour v.s. C, P<0.01; 4th hour v.s. C, P<0.01; 6th hour v.s. C, P<0.05), and returned to normal after 24 hours (24th hour v.s. C, P>0.99). \u0000Conclusions Exercise induced the distribution of HIF-1α in the small intestine. The expression of HIF-1α is shown in a time-dependent manner after exercise.","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"45 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90984316","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective A signal transduction pathway is always signal-specific, but it becomes special in exercise physiology, especially for habitual exercise training. In this paper, the normal signal transduction pathways (NPs) of habitual endurance exercise of Sprague-Dawly (SD) rats at different intensities were studied in terms of self-similar algorithm . Methods 1. SD rats were randomly divided into a control group with no exercise (Cont), a low-intensity exercise group (LI) at 15 m/min for 60 min, a medium-intensity exercise group (MI) at 25 m/min for 40 min, and a high-intensity exercise group (HI) 35 m/min for 30 min. The the treadmill exercise began at 10 m/min for 10 min. The speed and time increased by 5 m/min and by 5 min respectively after every two days until the intensity was OK for LI, MI and HI groups. All rats in the exercise groups were trained for eight weeks. The muscle was collected two hours after the last training. The 20 parameters, the expression of PGC-1α, PI3K, AMPK, p38MAPK, SIRT1, IGF-1, Akt, mTOR, TFAM, NRF1 and NRF2 and the level of T, CS, SDH, β-HAD, MDH, HK, PK, LDH and HSL, were assessed. 2. The 20 dimension data sets were integratedly analyzed with self-similar algorithm. Results 1. The first-order self-similarity held for the 20 dimension data sets between MI and HI groups. 2. There was one biomarker, PGC-1α, between Cont and LI groups with respect to HI group which parameters were used to be dimensionless for Cont and LI groups. 3. There were two biomarkers, PGC-1α and PI3K, between LI and MI or HI groups with respect cont group. Conclusions The NP of LI group may be PGC-1α pathway, and the NPs of MI or HI group may be PGC-1α and PI3K pathways.
{"title":"PO-239 Normal signal transduction pathways of rat habitual endurance exercise at different intensities","authors":"T. Liu, Jing-Gang Chen, Rui Duan, Quanguang Zhang","doi":"10.14428/EBR.V1I5.10873","DOIUrl":"https://doi.org/10.14428/EBR.V1I5.10873","url":null,"abstract":"Objective A signal transduction pathway is always signal-specific, but it becomes special in exercise physiology, especially for habitual exercise training. In this paper, the normal signal transduction pathways (NPs) of habitual endurance exercise of Sprague-Dawly (SD) rats at different intensities were studied in terms of self-similar algorithm . \u0000Methods 1. SD rats were randomly divided into a control group with no exercise (Cont), a low-intensity exercise group (LI) at 15 m/min for 60 min, a medium-intensity exercise group (MI) at 25 m/min for 40 min, and a high-intensity exercise group (HI) 35 m/min for 30 min. The the treadmill exercise began at 10 m/min for 10 min. The speed and time increased by 5 m/min and by 5 min respectively after every two days until the intensity was OK for LI, MI and HI groups. All rats in the exercise groups were trained for eight weeks. The muscle was collected two hours after the last training. The 20 parameters, the expression of PGC-1α, PI3K, AMPK, p38MAPK, SIRT1, IGF-1, Akt, mTOR, TFAM, NRF1 and NRF2 and the level of T, CS, SDH, β-HAD, MDH, HK, PK, LDH and HSL, were assessed. 2. The 20 dimension data sets were integratedly analyzed with self-similar algorithm. \u0000Results 1. The first-order self-similarity held for the 20 dimension data sets between MI and HI groups. 2. There was one biomarker, PGC-1α, between Cont and LI groups with respect to HI group which parameters were used to be dimensionless for Cont and LI groups. 3. There were two biomarkers, PGC-1α and PI3K, between LI and MI or HI groups with respect cont group. \u0000Conclusions The NP of LI group may be PGC-1α pathway, and the NPs of MI or HI group may be PGC-1α and PI3K pathways.","PeriodicalId":12276,"journal":{"name":"Exercise Biochemistry Review","volume":"86 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90177084","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：