The endochondral ossification of hindlimb is essential to a bird’s ability to stand,walk and fly.Most hindlimb is ossified in the embryos before hatching in precocial birds.However,the molecular mechanisms of hindlim...The endochondral ossification of hindlimb is essential to a bird’s ability to stand,walk and fly.Most hindlimb is ossified in the embryos before hatching in precocial birds.However,the molecular mechanisms of hindlimb ossification in birds is still unclear.Therefore,we tried to examine the process of hindlimb ossification and its molecular regulation by using an animal model—Japanese Quail(Coturnix japonica).We selected four critical stages(Embryo Day:E6,E8,E12 and E16) of skeletal development of embryonic quails for hindlimb skeleton staining to show the process of endochondral ossification and to examine the molecular regulation of endochondral osteogenesis by RNA-Seq analysis.The results showed that ossification became increased with embryonic development and most hindlimb was ossified before hatching.RNA-Seq analysis revealed that various signaling pathways were involved with endochondral ossification with thyroid hormone signaling and WNT signaling pathway particularly enriched.Moreover,the expression levels of 42 genes were continuously upregulated and 14 genes were continuously downregulated from E6 to E16.The present study might provide new insights into complex molecular mechanisms in regulation of endochondral ossification.展开更多
AIM To evaluate the angiogenic effect of platelet-rich plasma(PRP)-preconditioned adipose-derived stem cells(ADSCs) both in vitro and in a mouse ischemic hindlimb model.METHODS ADSCs were divided based on culture medi...AIM To evaluate the angiogenic effect of platelet-rich plasma(PRP)-preconditioned adipose-derived stem cells(ADSCs) both in vitro and in a mouse ischemic hindlimb model.METHODS ADSCs were divided based on culture medium: 2.5% PRP, 5% PRP, 7.5% PRP, and 10% PRP. Cell proliferation rate was analyzed using the MTS assay. The gene expression of CD31, vascular endothelial growth factor, hypoxia-inducible factors, and endothelial cell nitric oxide synthase was analyzed using reverse transcription polymerase chain reaction. Cell markers and structural changes were assessed through immunofluorescence staining and the tube formation assay. Subsequently, we studied the in vivo angiogenic capabilities of ADSCs by a mouse ischemic hindlimb model.RESULTS The proliferation rate of ADSCs was higher in the 2.5%, 5%, and 7.5% PRP groups. The expression of hypoxia-inducible factor, CD31, vascular endothelial growth factor, and endothelial cell nitric oxide synthase in the 5% and 7.5% PRP groups increased. The 5%, 7.5%, and 10% PRP groups showed higher abilities to promote both CD31 and vascular endothelial growth factor production and tubular structure formation in ADSCs. According to laser Doppler perfusion scan, the perfusion ratios of ischemic limb to normal limb were significantly higher in 5% PRP, 7.5% PRP, and human umbilical vein endothelial cells groups compared with the negative control and fetal bovine serum(FBS) groups(0.88 ± 0.08, 0.85 ± 0.07 and 0.81 ± 0.06 for 5%, 7.5% PRP and human umbilical vein endothelial cells compared with 0.42 ± 0.17 and 0.54 ± 0.14 for the negative control and FBS, P < 0.01).CONCLUSION PRP-preconditioned ADSCs presented endothelial cell characteristics in vitro and significantly improved neovascularization in ischemic hindlimbs. The optimal angiogenic effect occurred in 5% PRP-and 7.5% PRPpreconditioned ADSCs.展开更多
In many anurans, the forelimb muscles of males are used to grasp females and are often heavier than those of females despite the larger female body size. Such sexual dimorphism in forelimb musculature is thought to re...In many anurans, the forelimb muscles of males are used to grasp females and are often heavier than those of females despite the larger female body size. Such sexual dimorphism in forelimb musculature is thought to result from sexual selection. In addition, the hindlimbs of frogs and toads play an important role in the reproductive process as amplectant males can expel rivals with robust hindlimbs through kicking. In this study, the sexual dimorphism in dry mass for six hindlimb muscles of the Asiatic toad （Bufo gargarizans） was investigated. The results showed that, when controlled for body size, the hindlimb muscle mass of males significantly exceeded that of females for every muscle. The hindlimb muscle mass of amplectant males was also significantly larger than that of non-amplectant males. These results suggested that if strong hindlimb muscles could improve mating success of males, sexual selection would promote the evolution of dimorphism in this character.展开更多
Hindlimb unloading(HU)in rodent is a well-accepted ground-based model used to simulate some of the conditions of space flight and reproduce its deleterious effects on the musculoskeletal,cardiovascular and immune syst...Hindlimb unloading(HU)in rodent is a well-accepted ground-based model used to simulate some of the conditions of space flight and reproduce its deleterious effects on the musculoskeletal,cardiovascular and immune systems.In this study,the effects of HU on lymphocyte homeostasis in the spleen and thymus of mice were examined.HU was found to drastically deplete various cell populations in the spleen and thymus.These changes are likely to be mediated by apoptosis,since DNA strand breaks indicative of apoptosis were detected by terminal deoxynucleotidyl transferase-mediated nick end-labeling in both splenocytes and thymocytes.Surprisingly,administration of opioid antagonists or interference with the Fas-FasL interaction was able to block HU-induced reductions of splenocytes,but not thymocytes.On the other hand,steroid receptor antagonists blocked the reduction of lymphocyte numbers in both spleen and thymus.Therefore,the effects of HU on the homeostasis of splenocytes and thymocytes must be exerted through distinct mechanisms.展开更多
Assessment of locomotion recovery in preclinical studies of experimental spinal cord injury remains challenging. We studied the CatWalk XT■gait analysis for evaluating hindlimb functional recovery in a widely used an...Assessment of locomotion recovery in preclinical studies of experimental spinal cord injury remains challenging. We studied the CatWalk XT■gait analysis for evaluating hindlimb functional recovery in a widely used and clinically relevant thoracic contusion/compression spinal cord injury model in rats. Rats were randomly assigned to either a T9 spinal cord injury or sham laminectomy. Locomotion recovery was assessed using the Basso, Beattie, and Bresnahan open field rating scale and the CatWalk XT■gait analysis. To determine the potential bias from weight changes, corrected hindlimb(H) values(divided by the unaffected forelimb(F) values) were calculated. Six weeks after injury, cyst formation, astrogliosis, and the deposition of chondroitin sulfate glycosaminoglycans were assessed by immunohistochemistry staining. Compared with the baseline, a significant spontaneous recovery could be observed in the CatWalk XT■parameters max intensity, mean intensity, max intensity at%, and max contact mean intensity from 4 weeks after injury onwards. Of note, corrected values(H/F) of CatWalk XT■parameters showed a significantly less vulnerability to the weight changes than absolute values, specifically in static parameters. The corrected CatWalk XT■parameters were positively correlated with the Basso, Beattie, and Bresnahan rating scale scores, cyst formation, the immunointensity of astrogliosis and chondroitin sulfate glycosaminoglycan deposition. The CatWalk XT■gait analysis and especially its static parameters, therefore, seem to be highly useful in assessing spontaneous recovery of hindlimb function after severe thoracic spinal cord injury. Because many CatWalk XT■parameters of the hindlimbs seem to be affected by body weight changes, using their corrected values might be a valuable option to improve this dependency.展开更多
Male-biased sexual dimorphism in hind limb muscles is widespread in anuran species where scramble competition is common among males. Such sexual difference is thought to result from sexual selection. In this view, we ...Male-biased sexual dimorphism in hind limb muscles is widespread in anuran species where scramble competition is common among males. Such sexual difference is thought to result from sexual selection. In this view, we tested the differences in muscle mass between the sexes and between amplectant and non-amplectant males by quantifying the mass of four hindlimb muscles （triceps femoris, sartorius, gracilis and plantaris longus） of females and males of Odorrana schmackeri. The results showed that females significantly exceeded males for muscle triceps femoris, gracilis, plantaris longus and total mass when controlled for body size. There are no significant differences between amplectant and non-amplectant males. It is probable that the maintenance of the amplectant position in O. schmackeri may depend on the strength of hindlimb muscles in females to support the pair.展开更多
Reintroduction is an important strategy to restore or re-establish wild populations of endangered species.Pre-release training is a necessary step to ensure postreintroduction survival.However,studies reported contrad...Reintroduction is an important strategy to restore or re-establish wild populations of endangered species.Pre-release training is a necessary step to ensure postreintroduction survival.However,studies reported contradicting outcomes after pre-release training of juveniles and adults.This study used farmed and feral American mink(Neovison vison)to analyze the influence of captive breeding on the morphology,structure and efficiency of the two major hindlimb levers,the femur and tibia pivoted by hip and knee joints that are essential for locomotion.Results showed that captive breeding did not alter the sexual dimorphism of the two levers that are related to survival in the wild.Captive-bred mink showed slightly altered morphology of the femur and fundamental structure of the hindlimb levers that improved efficiency,but this resulted in reduction of performance related to foraging in both terrestrial and aquatic environments,especially for females.These findings suggest that reintroduction of mustelid as exampled by the mink here should focus on juveniles because the skeletal alterations associated with captive rearing were recorded only among adults and are irreversible in adulthood.In contrast,captive-reared juveniles showed no skeletal alterations and would be expected to recovery from any atrophy of the muscular system caused by captive rearing for shorter durations.Our results support the application of pre-release training of juveniles in enriched environments as a method for alleviating structural alteration of appendages and enhancing locomotion to increase survival probability in complex habitats.展开更多
The macrophage-mediated inflammatory response is crucial for the recovery of skeletal muscle following ischemia.Therefore,macrophage-based therapeutic targets need to be explored for ischemic disease.In the current st...The macrophage-mediated inflammatory response is crucial for the recovery of skeletal muscle following ischemia.Therefore,macrophage-based therapeutic targets need to be explored for ischemic disease.In the current study,we found that the mRNA levels of scavenger receptor A1(Sr-a1)were elevated in patients with critical limb ischemia,based on an analysis of the Gene Expression Omnibus data.We then investigated the role and underlying mechanisms of macrophage SR-A1 in a mouse hindlimb ischemia(HLI)model.Compared with the Sr-a1^(fl/fl)mice,the Lyz^(Cre+)/Sr-a1^(flox/flox)(Sr-a1~(ΔMΦ))mice showed significantly reduced laser Doppler blood flow in the ischemic limb on day seven after HLI.Consistently,histological analysis revealed that the ischemic limb of the Sr-a1~(ΔMΦ)mice exhibited more severe and prolonged necrotic morphology,inflammation,fibrosis,decreased vessel density,and delayed regeneration than that of the control Sr-a1~(fl/fl)mice.Furthermore,restoring wild-type myeloid cells to the Sr-a1 knockout mice effectively improved the Doppler perfusion in the ischemic limb and mitigated skeletal muscle damage seven days after HLI.Consistent with these in vivo findings,co-cultivating macrophages with the mouse myoblast cell line C2C12 revealed that the Sr-a1^(-/-)bone marrow macrophages significantly inhibited myoblast differentiation in vitro.Mechanistically,SR-A1 enhanced the skeletal muscle regeneration in response to HLI by inhibiting oncostatin M production via suppression of the NF-κB signaling activation.These findings indicate that SR-A1 may be a promising candidate protein to improve tissue repair and regeneration in peripheral ischemic arterial disease.展开更多
Objective:To investigate the effect and mechanism of Xuefu Zhuyu Capsule(血府逐瘀胶囊,XZC)on pro-angiogenesis in the hindlimb ischemic model rats.Methods:A total of 100 Sprague Dawley rats were randomly divided into a...Objective:To investigate the effect and mechanism of Xuefu Zhuyu Capsule(血府逐瘀胶囊,XZC)on pro-angiogenesis in the hindlimb ischemic model rats.Methods:A total of 100 Sprague Dawley rats were randomly divided into a model group,a regular-dose XZC group(0.48 g·kg^-1·d^-1)and a high-dose XZC group(0.96 g·kg^-1·d^-1)using random number table method.The model of hindlimb ischemic rats were made through femoral artery embolization with Bletilla microsphere age nt.XZC were give n on the first day after embolization surgery and lasted 5 days.Finally 72 models were obtained with 12 in each group for each time point.The lower Ischemic limb was amputated on the third day after embolization surgery.Histopathological characters and the number of blood vessels of granulation tissues were observed at 36 and 48 h after amputation,respectively.The main genes were obtained from microarray analysis and were validated using real-time quarttitative polymerase chain reaction.Results:The vascular number of granulation tissues at both 36 and 48 h were characterized by new and fresh vessels.The number of angiogenesis in the high-dose XZC group at 36 and 48 h was greater compared with that in the regular-dose XZC and model groups(P<0.01),and high-dose XZC at 36 h increased more vessels than that at 48 h(P<0.01).Consequently,granulation tissues from the high-dose XZC group at 36 h were chosen for microarray analysis.In all,2,085 differentially expressed genes(DEGs)were detected and 25 DEGs were determined to be directly related to an giogenesis.Four biological process terms were found including an giogenesis,regulati on of an gioge nesis,positive regulati on of an giogenesis,and positive regulation of vascular end othelial growth factor receptor sign aling pathway(P<0.05).Microarray an alysis also showed 49 pathways including 11 pathways related to an giogenesis.Conclusion:XZC promoted angiogenesis moderately and the mechanism invoIved multiple DEGs and multiple pathways.展开更多
Background Our previous research has suggested that platelet activating factor receptor was related to atherosclerosis. The present study investigated the effect of a platelet activating factor receptor antagonist- WE...Background Our previous research has suggested that platelet activating factor receptor was related to atherosclerosis. The present study investigated the effect of a platelet activating factor receptor antagonist- WEB2086 on angiogenesis in aortal plaque and ischemic hindlimb of apolipoprotein E-deficient mice. Methods Eight-week-old apolipoprotein E-deficient mice were fed with a 0.15% cholesterol diet to develop advanced lesions. At age 32 weeks unilateral hindlimb ischemia was surgically induced and the mice were divided into two groups： with or without WEB2086 mixed with their drinking water （4.3 mg in 100 ml）. At age 40 weeks blood was collected from the orbit for measurement of serum lipids and an enzyme linked immunosorbent assay was used to determine platelet activating factor and oxidized low density lipoprotein in the gastrocnemius and aorta. Whole-Mount CD31 stain and plaque-associated sprouting have been used to estimate angiogenesis in plaque from the aorta and laser Doppler perfusion imaging and immunohistochemical expression of von Willebrand factor have been used to estimate angiogenesis in ischemic hindlimb. Results The lipid composition of serum was not different between the groups. However, the amount of platelet activating factor and oxidized low density lipoprotein detected in the aorta was significantly higher than that in the gastrocnemius of ischemic hindlimb. The ratio of lesion to aorta levels was significantly reduced by administration of WEB2086, （31.52±6.18）% vs （55.58±8.34）%, P〈0.01. The mean density of intimal capillaries in atherosclerotic plaque, （31.13±9.20）% vs （57.74±11.28）%, P〈0.01, and the mean number of sprouts per aorta were significantly reduced, 183.92±34.17 vs 392.54±76.79, P〈0.01, in the WEB2086 group. Blood flow （0.85±0.12 vs 0.45±0.06, P〈0.01） and capillary density of ischemic hindlimb （1.18±0.17 vs 0.53±0.09, P〈0.01） were markedly increased in apolipoprotein E-deficient mice treated with WEB2086 versus controls. Conclusion The study provides evidence that WEB2086 can inhibit angiogenesis in atherosclerotic plaque but promote it in ischemic hindlimb.展开更多
目的:NOD样受体热蛋白结构域相关蛋白6(NOD-like receptor thermal protein domain associated protein 6,NLRP6)是新发现的寡聚化核苷酸结合结构域样受体家族成员,广泛表达于肠道、肝脏、肾脏、脾脏和肌肉等组织器官,在炎症、焦亡和自...目的:NOD样受体热蛋白结构域相关蛋白6(NOD-like receptor thermal protein domain associated protein 6,NLRP6)是新发现的寡聚化核苷酸结合结构域样受体家族成员,广泛表达于肠道、肝脏、肾脏、脾脏和肌肉等组织器官,在炎症、焦亡和自噬等多种生物过程发挥广泛的调节作用。近期研究表明,NLRP6在应激条件下对多种组织器官的疾病表型具有重要影响。然而,NLRP6对组织器官生长发育的影响尚未可知。方法:采用CRISPR/Cas9基因编辑技术构建NLRP6基因敲除小鼠,饲养并记录小鼠的生长和繁殖情况。将小鼠的脾、肝、心、肾、脑、四肢和后背皮肤等组织器官进行解剖和切片染色,以评估NLRP6基因敲除对实质器官的宏观发育影响和对组织结构的微观影响。结果:在自然状态下,NLRP6基因敲除缩短了雄鼠的性成熟期并使成年雄鼠的睾丸发生不可逆的破溃与萎缩。在四肢发育上,NLRP6基因敲除诱导成年雄鼠后肢横纹肌断裂,导致后肢明显萎缩。在脾脏发育上,NLRP6基因敲除不仅显著增加了雄鼠的脾脏体积(P<0.01)还诱导雄鼠脾脏出现炎性细胞浸润。在后背皮肤上,NLRP6基因敲除引发雄鼠后背皮肤出现明显的溃疡损伤、胶原纤维增生和炎性细胞浸润。结论:在自然生长发育条件下,NLRP6基因敲除选择性影响小鼠的生殖器发育与性成熟期、后肢肌肉发育、脾脏大小及其炎症免疫状态以及后背皮肤的结构完整性,而且这一作用具有明显的雄激素依赖性。展开更多
基金supported by the National Natural Science Foundationof China (No. 33372201)。
文摘The endochondral ossification of hindlimb is essential to a bird’s ability to stand,walk and fly.Most hindlimb is ossified in the embryos before hatching in precocial birds.However,the molecular mechanisms of hindlimb ossification in birds is still unclear.Therefore,we tried to examine the process of hindlimb ossification and its molecular regulation by using an animal model—Japanese Quail(Coturnix japonica).We selected four critical stages(Embryo Day:E6,E8,E12 and E16) of skeletal development of embryonic quails for hindlimb skeleton staining to show the process of endochondral ossification and to examine the molecular regulation of endochondral osteogenesis by RNA-Seq analysis.The results showed that ossification became increased with embryonic development and most hindlimb was ossified before hatching.RNA-Seq analysis revealed that various signaling pathways were involved with endochondral ossification with thyroid hormone signaling and WNT signaling pathway particularly enriched.Moreover,the expression levels of 42 genes were continuously upregulated and 14 genes were continuously downregulated from E6 to E16.The present study might provide new insights into complex molecular mechanisms in regulation of endochondral ossification.
基金Supported by grant from the National Sci-Tech Program,Ministry of Science and Technology,No.NRMPG3E0471 and No.NMRPG3D0231a Chang Gung Memorial Hospital grant,No.CMRPGBH0011
文摘AIM To evaluate the angiogenic effect of platelet-rich plasma(PRP)-preconditioned adipose-derived stem cells(ADSCs) both in vitro and in a mouse ischemic hindlimb model.METHODS ADSCs were divided based on culture medium: 2.5% PRP, 5% PRP, 7.5% PRP, and 10% PRP. Cell proliferation rate was analyzed using the MTS assay. The gene expression of CD31, vascular endothelial growth factor, hypoxia-inducible factors, and endothelial cell nitric oxide synthase was analyzed using reverse transcription polymerase chain reaction. Cell markers and structural changes were assessed through immunofluorescence staining and the tube formation assay. Subsequently, we studied the in vivo angiogenic capabilities of ADSCs by a mouse ischemic hindlimb model.RESULTS The proliferation rate of ADSCs was higher in the 2.5%, 5%, and 7.5% PRP groups. The expression of hypoxia-inducible factor, CD31, vascular endothelial growth factor, and endothelial cell nitric oxide synthase in the 5% and 7.5% PRP groups increased. The 5%, 7.5%, and 10% PRP groups showed higher abilities to promote both CD31 and vascular endothelial growth factor production and tubular structure formation in ADSCs. According to laser Doppler perfusion scan, the perfusion ratios of ischemic limb to normal limb were significantly higher in 5% PRP, 7.5% PRP, and human umbilical vein endothelial cells groups compared with the negative control and fetal bovine serum(FBS) groups(0.88 ± 0.08, 0.85 ± 0.07 and 0.81 ± 0.06 for 5%, 7.5% PRP and human umbilical vein endothelial cells compared with 0.42 ± 0.17 and 0.54 ± 0.14 for the negative control and FBS, P < 0.01).CONCLUSION PRP-preconditioned ADSCs presented endothelial cell characteristics in vitro and significantly improved neovascularization in ischemic hindlimbs. The optimal angiogenic effect occurred in 5% PRP-and 7.5% PRPpreconditioned ADSCs.
基金supported by the National Natural Science Foundation of China(31101633)the Innovative Team Foundation of China West Normal University
文摘In many anurans, the forelimb muscles of males are used to grasp females and are often heavier than those of females despite the larger female body size. Such sexual dimorphism in forelimb musculature is thought to result from sexual selection. In addition, the hindlimbs of frogs and toads play an important role in the reproductive process as amplectant males can expel rivals with robust hindlimbs through kicking. In this study, the sexual dimorphism in dry mass for six hindlimb muscles of the Asiatic toad （Bufo gargarizans） was investigated. The results showed that, when controlled for body size, the hindlimb muscle mass of males significantly exceeded that of females for every muscle. The hindlimb muscle mass of amplectant males was also significantly larger than that of non-amplectant males. These results suggested that if strong hindlimb muscles could improve mating success of males, sexual selection would promote the evolution of dimorphism in this character.
文摘Hindlimb unloading(HU)in rodent is a well-accepted ground-based model used to simulate some of the conditions of space flight and reproduce its deleterious effects on the musculoskeletal,cardiovascular and immune systems.In this study,the effects of HU on lymphocyte homeostasis in the spleen and thymus of mice were examined.HU was found to drastically deplete various cell populations in the spleen and thymus.These changes are likely to be mediated by apoptosis,since DNA strand breaks indicative of apoptosis were detected by terminal deoxynucleotidyl transferase-mediated nick end-labeling in both splenocytes and thymocytes.Surprisingly,administration of opioid antagonists or interference with the Fas-FasL interaction was able to block HU-induced reductions of splenocytes,but not thymocytes.On the other hand,steroid receptor antagonists blocked the reduction of lymphocyte numbers in both spleen and thymus.Therefore,the effects of HU on the homeostasis of splenocytes and thymocytes must be exerted through distinct mechanisms.
文摘Assessment of locomotion recovery in preclinical studies of experimental spinal cord injury remains challenging. We studied the CatWalk XT■gait analysis for evaluating hindlimb functional recovery in a widely used and clinically relevant thoracic contusion/compression spinal cord injury model in rats. Rats were randomly assigned to either a T9 spinal cord injury or sham laminectomy. Locomotion recovery was assessed using the Basso, Beattie, and Bresnahan open field rating scale and the CatWalk XT■gait analysis. To determine the potential bias from weight changes, corrected hindlimb(H) values(divided by the unaffected forelimb(F) values) were calculated. Six weeks after injury, cyst formation, astrogliosis, and the deposition of chondroitin sulfate glycosaminoglycans were assessed by immunohistochemistry staining. Compared with the baseline, a significant spontaneous recovery could be observed in the CatWalk XT■parameters max intensity, mean intensity, max intensity at%, and max contact mean intensity from 4 weeks after injury onwards. Of note, corrected values(H/F) of CatWalk XT■parameters showed a significantly less vulnerability to the weight changes than absolute values, specifically in static parameters. The corrected CatWalk XT■parameters were positively correlated with the Basso, Beattie, and Bresnahan rating scale scores, cyst formation, the immunointensity of astrogliosis and chondroitin sulfate glycosaminoglycan deposition. The CatWalk XT■gait analysis and especially its static parameters, therefore, seem to be highly useful in assessing spontaneous recovery of hindlimb function after severe thoracic spinal cord injury. Because many CatWalk XT■parameters of the hindlimbs seem to be affected by body weight changes, using their corrected values might be a valuable option to improve this dependency.
基金supported by National Sciences Foundation of China (No. 31372164, 30870277)the Scientific Research Foundation of Henan Normal University (No. 01046500145)+1 种基金Joint Funds for Fostering Talents of NSFC the People’s Government of Henan Province (Grant No. U1304309)
文摘Male-biased sexual dimorphism in hind limb muscles is widespread in anuran species where scramble competition is common among males. Such sexual difference is thought to result from sexual selection. In this view, we tested the differences in muscle mass between the sexes and between amplectant and non-amplectant males by quantifying the mass of four hindlimb muscles （triceps femoris, sartorius, gracilis and plantaris longus） of females and males of Odorrana schmackeri. The results showed that females significantly exceeded males for muscle triceps femoris, gracilis, plantaris longus and total mass when controlled for body size. There are no significant differences between amplectant and non-amplectant males. It is probable that the maintenance of the amplectant position in O. schmackeri may depend on the strength of hindlimb muscles in females to support the pair.
基金funded by China State Forestry and Grassland Administration Project for Rescue and Captive Breeding of Endangered and Rare Wildlife(2018).
文摘Reintroduction is an important strategy to restore or re-establish wild populations of endangered species.Pre-release training is a necessary step to ensure postreintroduction survival.However,studies reported contradicting outcomes after pre-release training of juveniles and adults.This study used farmed and feral American mink(Neovison vison)to analyze the influence of captive breeding on the morphology,structure and efficiency of the two major hindlimb levers,the femur and tibia pivoted by hip and knee joints that are essential for locomotion.Results showed that captive breeding did not alter the sexual dimorphism of the two levers that are related to survival in the wild.Captive-bred mink showed slightly altered morphology of the femur and fundamental structure of the hindlimb levers that improved efficiency,but this resulted in reduction of performance related to foraging in both terrestrial and aquatic environments,especially for females.These findings suggest that reintroduction of mustelid as exampled by the mink here should focus on juveniles because the skeletal alterations associated with captive rearing were recorded only among adults and are irreversible in adulthood.In contrast,captive-reared juveniles showed no skeletal alterations and would be expected to recovery from any atrophy of the muscular system caused by captive rearing for shorter durations.Our results support the application of pre-release training of juveniles in enriched environments as a method for alleviating structural alteration of appendages and enhancing locomotion to increase survival probability in complex habitats.
基金supported by the National Natural Science Foundation of China(Grant Nos.82030012,81670263,82170444,82270476,82270361,and 82100433)the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(Grant No.24KJA310003)。
文摘The macrophage-mediated inflammatory response is crucial for the recovery of skeletal muscle following ischemia.Therefore,macrophage-based therapeutic targets need to be explored for ischemic disease.In the current study,we found that the mRNA levels of scavenger receptor A1(Sr-a1)were elevated in patients with critical limb ischemia,based on an analysis of the Gene Expression Omnibus data.We then investigated the role and underlying mechanisms of macrophage SR-A1 in a mouse hindlimb ischemia(HLI)model.Compared with the Sr-a1^(fl/fl)mice,the Lyz^(Cre+)/Sr-a1^(flox/flox)(Sr-a1~(ΔMΦ))mice showed significantly reduced laser Doppler blood flow in the ischemic limb on day seven after HLI.Consistently,histological analysis revealed that the ischemic limb of the Sr-a1~(ΔMΦ)mice exhibited more severe and prolonged necrotic morphology,inflammation,fibrosis,decreased vessel density,and delayed regeneration than that of the control Sr-a1~(fl/fl)mice.Furthermore,restoring wild-type myeloid cells to the Sr-a1 knockout mice effectively improved the Doppler perfusion in the ischemic limb and mitigated skeletal muscle damage seven days after HLI.Consistent with these in vivo findings,co-cultivating macrophages with the mouse myoblast cell line C2C12 revealed that the Sr-a1^(-/-)bone marrow macrophages significantly inhibited myoblast differentiation in vitro.Mechanistically,SR-A1 enhanced the skeletal muscle regeneration in response to HLI by inhibiting oncostatin M production via suppression of the NF-κB signaling activation.These findings indicate that SR-A1 may be a promising candidate protein to improve tissue repair and regeneration in peripheral ischemic arterial disease.
基金Supported by the National Natural Science Foundation of China(No.81072933,81173431)the Beijing Natural Science Foundation Program(No.7122120)
文摘Objective:To investigate the effect and mechanism of Xuefu Zhuyu Capsule(血府逐瘀胶囊,XZC)on pro-angiogenesis in the hindlimb ischemic model rats.Methods:A total of 100 Sprague Dawley rats were randomly divided into a model group,a regular-dose XZC group(0.48 g·kg^-1·d^-1)and a high-dose XZC group(0.96 g·kg^-1·d^-1)using random number table method.The model of hindlimb ischemic rats were made through femoral artery embolization with Bletilla microsphere age nt.XZC were give n on the first day after embolization surgery and lasted 5 days.Finally 72 models were obtained with 12 in each group for each time point.The lower Ischemic limb was amputated on the third day after embolization surgery.Histopathological characters and the number of blood vessels of granulation tissues were observed at 36 and 48 h after amputation,respectively.The main genes were obtained from microarray analysis and were validated using real-time quarttitative polymerase chain reaction.Results:The vascular number of granulation tissues at both 36 and 48 h were characterized by new and fresh vessels.The number of angiogenesis in the high-dose XZC group at 36 and 48 h was greater compared with that in the regular-dose XZC and model groups(P<0.01),and high-dose XZC at 36 h increased more vessels than that at 48 h(P<0.01).Consequently,granulation tissues from the high-dose XZC group at 36 h were chosen for microarray analysis.In all,2,085 differentially expressed genes(DEGs)were detected and 25 DEGs were determined to be directly related to an giogenesis.Four biological process terms were found including an giogenesis,regulati on of an gioge nesis,positive regulati on of an giogenesis,and positive regulation of vascular end othelial growth factor receptor sign aling pathway(P<0.05).Microarray an alysis also showed 49 pathways including 11 pathways related to an giogenesis.Conclusion:XZC promoted angiogenesis moderately and the mechanism invoIved multiple DEGs and multiple pathways.
基金This work was supported by Scientific Research Fund of Hunan Provincial Education Department(No.05C457).
文摘Background Our previous research has suggested that platelet activating factor receptor was related to atherosclerosis. The present study investigated the effect of a platelet activating factor receptor antagonist- WEB2086 on angiogenesis in aortal plaque and ischemic hindlimb of apolipoprotein E-deficient mice. Methods Eight-week-old apolipoprotein E-deficient mice were fed with a 0.15% cholesterol diet to develop advanced lesions. At age 32 weeks unilateral hindlimb ischemia was surgically induced and the mice were divided into two groups： with or without WEB2086 mixed with their drinking water （4.3 mg in 100 ml）. At age 40 weeks blood was collected from the orbit for measurement of serum lipids and an enzyme linked immunosorbent assay was used to determine platelet activating factor and oxidized low density lipoprotein in the gastrocnemius and aorta. Whole-Mount CD31 stain and plaque-associated sprouting have been used to estimate angiogenesis in plaque from the aorta and laser Doppler perfusion imaging and immunohistochemical expression of von Willebrand factor have been used to estimate angiogenesis in ischemic hindlimb. Results The lipid composition of serum was not different between the groups. However, the amount of platelet activating factor and oxidized low density lipoprotein detected in the aorta was significantly higher than that in the gastrocnemius of ischemic hindlimb. The ratio of lesion to aorta levels was significantly reduced by administration of WEB2086, （31.52±6.18）% vs （55.58±8.34）%, P〈0.01. The mean density of intimal capillaries in atherosclerotic plaque, （31.13±9.20）% vs （57.74±11.28）%, P〈0.01, and the mean number of sprouts per aorta were significantly reduced, 183.92±34.17 vs 392.54±76.79, P〈0.01, in the WEB2086 group. Blood flow （0.85±0.12 vs 0.45±0.06, P〈0.01） and capillary density of ischemic hindlimb （1.18±0.17 vs 0.53±0.09, P〈0.01） were markedly increased in apolipoprotein E-deficient mice treated with WEB2086 versus controls. Conclusion The study provides evidence that WEB2086 can inhibit angiogenesis in atherosclerotic plaque but promote it in ischemic hindlimb.
文摘目的:NOD样受体热蛋白结构域相关蛋白6(NOD-like receptor thermal protein domain associated protein 6,NLRP6)是新发现的寡聚化核苷酸结合结构域样受体家族成员,广泛表达于肠道、肝脏、肾脏、脾脏和肌肉等组织器官,在炎症、焦亡和自噬等多种生物过程发挥广泛的调节作用。近期研究表明,NLRP6在应激条件下对多种组织器官的疾病表型具有重要影响。然而,NLRP6对组织器官生长发育的影响尚未可知。方法:采用CRISPR/Cas9基因编辑技术构建NLRP6基因敲除小鼠,饲养并记录小鼠的生长和繁殖情况。将小鼠的脾、肝、心、肾、脑、四肢和后背皮肤等组织器官进行解剖和切片染色,以评估NLRP6基因敲除对实质器官的宏观发育影响和对组织结构的微观影响。结果:在自然状态下,NLRP6基因敲除缩短了雄鼠的性成熟期并使成年雄鼠的睾丸发生不可逆的破溃与萎缩。在四肢发育上,NLRP6基因敲除诱导成年雄鼠后肢横纹肌断裂,导致后肢明显萎缩。在脾脏发育上,NLRP6基因敲除不仅显著增加了雄鼠的脾脏体积(P<0.01)还诱导雄鼠脾脏出现炎性细胞浸润。在后背皮肤上,NLRP6基因敲除引发雄鼠后背皮肤出现明显的溃疡损伤、胶原纤维增生和炎性细胞浸润。结论:在自然生长发育条件下,NLRP6基因敲除选择性影响小鼠的生殖器发育与性成熟期、后肢肌肉发育、脾脏大小及其炎症免疫状态以及后背皮肤的结构完整性,而且这一作用具有明显的雄激素依赖性。
