Background Mastitis not only deteriorates the composition or quality of milk,but also damages the health and pro-ductivity of dairy goats.Sulforaphane(SFN)is a phytochemical isothiocyanate compound with various pharma...Background Mastitis not only deteriorates the composition or quality of milk,but also damages the health and pro-ductivity of dairy goats.Sulforaphane(SFN)is a phytochemical isothiocyanate compound with various pharmacologi-cal effects such as anti-oxidant and anti-inflammatory.However,the effect of SFN on mastitis has yet to be elucidated.This study aimed to explore the anti-oxidant and anti-inflammatory effects and potential molecular mechanisms of SFN in lipopolysaccharide(LPS)-induced primary goat mammary epithelial cells(GMECs)and a mouse model of mastitis.Results In vitro,SFN downregulated the mRNA expression of inflammatory factors(tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6),inhibited the protein expression of inflammatory mediators(cyclooxygenase-2(COX2),and inducible nitric oxide synthase(iNOS))while suppressing nuclear factor kappa-B(NF-κB)activation in LPS-induced GMECs.Additionally,SFN exhibited an antioxidant effect by increasing Nrf2 expression and nuclear translocation,up-regulating antioxidant enzymes expression,and decreasing LPS-induced reactive oxygen species(ROS)produc-tion in GMECs.Furthermore,SFN pretreatment promoted the autophagy pathway,which was dependent on the increased Nrf2 level,and contributed significantly to the improved LPS-induced oxidative stress and inflammatory response.In vivo,SFN effectively alleviated histopathological lesions,suppressed the expression of inflammatory factors,enhanced immunohistochemistry staining of Nrf2,and amplified of LC3 puncta LPS-induced mastitis in mice.Mechanically,the in vitro and in vivo study showed that the anti-inflammatory and anti-oxidative stress effects of SFN were mediated by the Nrf2-mediated autophagy pathway in GMECs and a mouse model of mastitis.Conclusions These results indicate that the natural compound SFN has a preventive effect on LPS-induced inflam-mation through by regulating the Nrf2-mediated autophagy pathway in primary goat mammary epithelial cells and a mouse model of mastitis,which may improve prevention strategies for mastitis in dairy goats.展开更多
The enrichment and identification of human epidermal stem cells （EpSCs） are of paramount importance for both basic research and clinical application. Although several approaches for the enrichment of EpSCs have been...The enrichment and identification of human epidermal stem cells （EpSCs） are of paramount importance for both basic research and clinical application. Although several approaches for the enrichment of EpSCs have been established, enriching a pure population of viable EpSCs is still a challenging task. An improved approach is worth developing to enhance the purity and viability of EpSCs. Here we report that cell size combined with collagen type IV adhesiveness can be used in an improved approach to enrich pure and viable human EpSCs. We separated the rap- idly adherent keratinocytes into three populations that range in size from 5-7 μm （population A）, to 7-9 μm （population B）, to ≥9μm （population C） in diameter, and found that human putative EpSCs could be further enriched in population A with the smallest size. Among the three populations, population A displayed the highest density of plintegrin receptor, contained the highest percentage of cells in G0/G1 phase, showed the highest nucleus to cytoplasm ratio, and possessed the highest colony formation efficiency （CFE）. When injected into murine blastocysts, these cells participated in multi-tissue formation. More significantly, compared with a previous approach that sorted putative EpSCs according to pl-integrin antibody staining, the viability of the EpSCs enriched by the improved approach was significantly enhanced. Our results provide a putative strategy for the enrichment of human EpSCs, and encourage further study into the role of cell size in stem cell biology.展开更多
Current measures mainly focus on how melatonin reduces physiological heat stress in animals,but its effects on reproductive damage to male dairy goats have been neglected.This study aimed to determine the protective e...Current measures mainly focus on how melatonin reduces physiological heat stress in animals,but its effects on reproductive damage to male dairy goats have been neglected.This study aimed to determine the protective effect of melatonin on male reproduction during heat stress in dairy goats and to further explore its mechanisms.A natural heat stress model of Saanen dairy goats was used to assess testicular tissue damage 7days after heat stress and to examine semen quality changes during a spermatogenic cycle.RNA-seq,Western blot,RT–qPCR,and immunofluo-rescence staining were used to explore the mechanism by which melatonin protects against heat stress-induced reproductive damage and to validate the results.The data suggested that melatonin significantly alleviated the heat stress-induced decrease in sperm quality,protected varicose tubule structure,reduced the levels of heat shock proteins and apoptotic proteins and protected the spermatocytes and round spermatozoa,which are mainly affected by heat stress.RNA-seq results suggest that melatonin inhibits the PI3K/AKT signaling pathway,reduces the level of p-AKT,and promotes elevated BCL-2.In addition,melatonin treatment could upregulate the gene expression of MT2 which was downregulated by heat stress and improve the change in extracellular matrix components and restore serum testosterone levels.Our results suggest that melatonin can protect against testicular and spermatogenic cell damage and improve semen quality in male dairy goats under heat stress.This study provides an important reference for subsequent studies on the molecular mechanisms of melatonin in protecting male reproductive processes under heat stress and using exogenous melatonin to prevent heat stress.展开更多
Sheep is an important livestock species raised globally to produce meat,milk,wool,and other by-products.During the Neolithic Revolution,sheep were domesticated in the Fertile Crescent of Southwest Asia around 10,000 y...Sheep is an important livestock species raised globally to produce meat,milk,wool,and other by-products.During the Neolithic Revolution,sheep were domesticated in the Fertile Crescent of Southwest Asia around 10,000 years ago(Chessa et al.,2009).展开更多
Early weaning induces intestinal injury,leading to a series of long-term symptoms such as inflammation,malabsorption and diarrhea.In this study,we hypothesized that microbes and theirmetabolitesmodulate the host's...Early weaning induces intestinal injury,leading to a series of long-term symptoms such as inflammation,malabsorption and diarrhea.In this study,we hypothesized that microbes and theirmetabolitesmodulate the host's inflammatory response to early weaning stress in a goatmodel.A total of 18 female Tibetan goat kids(n?9)wereweaned fromtheirmothers at 28 d(D28)and 60 d(D60)postpartum.D60 and D28 groupswere fed the same solid diet ad libitum fromweaning to 75 d of age.The colonic epithelium was subject to RNAsequencing,the caecal digesta metabolomics were assessed by liquid chromatographyetandem mass spectrometry(LC-MS/MS),and the caecal microbiota composition was analysed by 16S ribosomal RNA gene sequencing.We foundthatearlyweaningsubstantially increased the colonic pro-apoptotic gene expressionof B-cell lymphoma associated X(Bax),caspase-9,and caspase-3,and decreased the expression of zonula occludens-1(ZO-1)and claudin-1(P<0.01).In addition,a significant Bacteroides acidifaciens enrichmentwas observed in the hindgut of early-weaned goats(P<0.01),which negatively correlated with lysophosphatidylcholine products.Similarly,the chemokine signaling,IL-17 signaling,and peroxisome proliferatorsactivated receptor(PPAR)signaling pathways were upregulated in the colonic mucosa of the early-weaned goats.By applying caecal microbiota transplantation from goats to defaunated C57/6J mice,we confirmed that caecalmicrobiota of D28 goat kids increased the relative abundance of B.acidifaciens and significantly upregulated the genes of Bax,G proteinecoupled receptor(GPR)109A,GPR 43,fatty acid binding protein 6,nuclear receptor subfamily 1 group H member 3,angiotensin converting enzyme 2,and IL-6 expression(P<0.05),and decreased ZO-1,and claudin-1 protein expression in the mice jejunum and colon(P<0.001).These results proposed that the hindgut microbiota andmetabolites mediate the barrier functionweakening duringearlyweaning,and the relative abundance of B.acidifacienswas negatively correlatedwiththe hindgut barrier gene expression.This studydemonstrateshowweaningstress canaffectkeyhostemicrobe interaction regulators in the hindgut,in a lysophosphatidylcholine dependent and independent manner.Furthermore,based on our mice data,these results are transferable to other mammal species.展开更多
Effects of antibiotic residues on methane production in anaerobic digestion are commonly studied using the following two antibiotic addition methods：（1） adding manure from animals that consume a diet containing ant...Effects of antibiotic residues on methane production in anaerobic digestion are commonly studied using the following two antibiotic addition methods：（1） adding manure from animals that consume a diet containing antibiotics, and（2） adding antibiotic-free animal manure spiked with antibiotics. This study used chlortetracycline（CTC） as a model antibiotic to examine the effects of the antibiotic addition method on methane production in anaerobic digestion under two different swine wastewater concentrations（0.55 and 0.22 mg CTC/g dry manure）. The results showed that CTC degradation rate in which manure was directly added at 0.55 mg CTC/g（HSPIKE treatment） was lower than the control values and the rest of the treatment groups. Methane production from the HSPIKEtreatment was reduced（p 〈 0.05） by 12% during the whole experimental period and 15% during the first 7 days. The treatments had no significant effect on the pH and chemical oxygen demand value of the digesters, and the total nitrogen of the0.55 mg CTC/kg manure collected from mediated swine was significantly higher than the other values. Therefore, different methane production under different antibiotic addition methods might be explained by the microbial activity and the concentrations of antibiotic intermediate products and metabolites. Because the primary entry route of veterinary antibiotics into an anaerobic digester is by contaminated animal manure, the most appropriate method for studying antibiotic residue effects on methane production may be using manure from animals that are given a particular antibiotic, rather than adding the antibiotic directly to the anaerobic digester.展开更多
基金supported by Fuping County Dairy Goat High-efficiency Breeding Technology R&D and Extension Application Project(No.K3380216101)the Dairy Goat High-efficiency Breeding Technology Research and Application Project(No.K4040121023).
文摘Background Mastitis not only deteriorates the composition or quality of milk,but also damages the health and pro-ductivity of dairy goats.Sulforaphane(SFN)is a phytochemical isothiocyanate compound with various pharmacologi-cal effects such as anti-oxidant and anti-inflammatory.However,the effect of SFN on mastitis has yet to be elucidated.This study aimed to explore the anti-oxidant and anti-inflammatory effects and potential molecular mechanisms of SFN in lipopolysaccharide(LPS)-induced primary goat mammary epithelial cells(GMECs)and a mouse model of mastitis.Results In vitro,SFN downregulated the mRNA expression of inflammatory factors(tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6),inhibited the protein expression of inflammatory mediators(cyclooxygenase-2(COX2),and inducible nitric oxide synthase(iNOS))while suppressing nuclear factor kappa-B(NF-κB)activation in LPS-induced GMECs.Additionally,SFN exhibited an antioxidant effect by increasing Nrf2 expression and nuclear translocation,up-regulating antioxidant enzymes expression,and decreasing LPS-induced reactive oxygen species(ROS)produc-tion in GMECs.Furthermore,SFN pretreatment promoted the autophagy pathway,which was dependent on the increased Nrf2 level,and contributed significantly to the improved LPS-induced oxidative stress and inflammatory response.In vivo,SFN effectively alleviated histopathological lesions,suppressed the expression of inflammatory factors,enhanced immunohistochemistry staining of Nrf2,and amplified of LC3 puncta LPS-induced mastitis in mice.Mechanically,the in vitro and in vivo study showed that the anti-inflammatory and anti-oxidative stress effects of SFN were mediated by the Nrf2-mediated autophagy pathway in GMECs and a mouse model of mastitis.Conclusions These results indicate that the natural compound SFN has a preventive effect on LPS-induced inflam-mation through by regulating the Nrf2-mediated autophagy pathway in primary goat mammary epithelial cells and a mouse model of mastitis,which may improve prevention strategies for mastitis in dairy goats.
文摘The enrichment and identification of human epidermal stem cells （EpSCs） are of paramount importance for both basic research and clinical application. Although several approaches for the enrichment of EpSCs have been established, enriching a pure population of viable EpSCs is still a challenging task. An improved approach is worth developing to enhance the purity and viability of EpSCs. Here we report that cell size combined with collagen type IV adhesiveness can be used in an improved approach to enrich pure and viable human EpSCs. We separated the rap- idly adherent keratinocytes into three populations that range in size from 5-7 μm （population A）, to 7-9 μm （population B）, to ≥9μm （population C） in diameter, and found that human putative EpSCs could be further enriched in population A with the smallest size. Among the three populations, population A displayed the highest density of plintegrin receptor, contained the highest percentage of cells in G0/G1 phase, showed the highest nucleus to cytoplasm ratio, and possessed the highest colony formation efficiency （CFE）. When injected into murine blastocysts, these cells participated in multi-tissue formation. More significantly, compared with a previous approach that sorted putative EpSCs according to pl-integrin antibody staining, the viability of the EpSCs enriched by the improved approach was significantly enhanced. Our results provide a putative strategy for the enrichment of human EpSCs, and encourage further study into the role of cell size in stem cell biology.
基金supported by the natural sciences foundation of China(32072815)the technology innovation leading program of Shaanxi province(2020QFY10)+1 种基金the program of Shaanxi province science and technology innovation team(2019TD-036)the fundamental research funds for the central universities(2452020157).
文摘Current measures mainly focus on how melatonin reduces physiological heat stress in animals,but its effects on reproductive damage to male dairy goats have been neglected.This study aimed to determine the protective effect of melatonin on male reproduction during heat stress in dairy goats and to further explore its mechanisms.A natural heat stress model of Saanen dairy goats was used to assess testicular tissue damage 7days after heat stress and to examine semen quality changes during a spermatogenic cycle.RNA-seq,Western blot,RT–qPCR,and immunofluo-rescence staining were used to explore the mechanism by which melatonin protects against heat stress-induced reproductive damage and to validate the results.The data suggested that melatonin significantly alleviated the heat stress-induced decrease in sperm quality,protected varicose tubule structure,reduced the levels of heat shock proteins and apoptotic proteins and protected the spermatocytes and round spermatozoa,which are mainly affected by heat stress.RNA-seq results suggest that melatonin inhibits the PI3K/AKT signaling pathway,reduces the level of p-AKT,and promotes elevated BCL-2.In addition,melatonin treatment could upregulate the gene expression of MT2 which was downregulated by heat stress and improve the change in extracellular matrix components and restore serum testosterone levels.Our results suggest that melatonin can protect against testicular and spermatogenic cell damage and improve semen quality in male dairy goats under heat stress.This study provides an important reference for subsequent studies on the molecular mechanisms of melatonin in protecting male reproductive processes under heat stress and using exogenous melatonin to prevent heat stress.
文摘Sheep is an important livestock species raised globally to produce meat,milk,wool,and other by-products.During the Neolithic Revolution,sheep were domesticated in the Fertile Crescent of Southwest Asia around 10,000 years ago(Chessa et al.,2009).
基金Qinghai Province Key R&D and Transformation Plan(2020-NK-127)Tibet Science and Technology Department's"13th Five-Year Plan"Major Agriculture Project(XZ201901NA02)+2 种基金the Key Technology R&D Program of Xinjiang Groups(2020AB016)Science and Technology Program of Tibet(XZ202001YD0017C)Agriculture Research System of China(CARS-39-12).
文摘Early weaning induces intestinal injury,leading to a series of long-term symptoms such as inflammation,malabsorption and diarrhea.In this study,we hypothesized that microbes and theirmetabolitesmodulate the host's inflammatory response to early weaning stress in a goatmodel.A total of 18 female Tibetan goat kids(n?9)wereweaned fromtheirmothers at 28 d(D28)and 60 d(D60)postpartum.D60 and D28 groupswere fed the same solid diet ad libitum fromweaning to 75 d of age.The colonic epithelium was subject to RNAsequencing,the caecal digesta metabolomics were assessed by liquid chromatographyetandem mass spectrometry(LC-MS/MS),and the caecal microbiota composition was analysed by 16S ribosomal RNA gene sequencing.We foundthatearlyweaningsubstantially increased the colonic pro-apoptotic gene expressionof B-cell lymphoma associated X(Bax),caspase-9,and caspase-3,and decreased the expression of zonula occludens-1(ZO-1)and claudin-1(P<0.01).In addition,a significant Bacteroides acidifaciens enrichmentwas observed in the hindgut of early-weaned goats(P<0.01),which negatively correlated with lysophosphatidylcholine products.Similarly,the chemokine signaling,IL-17 signaling,and peroxisome proliferatorsactivated receptor(PPAR)signaling pathways were upregulated in the colonic mucosa of the early-weaned goats.By applying caecal microbiota transplantation from goats to defaunated C57/6J mice,we confirmed that caecalmicrobiota of D28 goat kids increased the relative abundance of B.acidifaciens and significantly upregulated the genes of Bax,G proteinecoupled receptor(GPR)109A,GPR 43,fatty acid binding protein 6,nuclear receptor subfamily 1 group H member 3,angiotensin converting enzyme 2,and IL-6 expression(P<0.05),and decreased ZO-1,and claudin-1 protein expression in the mice jejunum and colon(P<0.001).These results proposed that the hindgut microbiota andmetabolites mediate the barrier functionweakening duringearlyweaning,and the relative abundance of B.acidifacienswas negatively correlatedwiththe hindgut barrier gene expression.This studydemonstrateshowweaningstress canaffectkeyhostemicrobe interaction regulators in the hindgut,in a lysophosphatidylcholine dependent and independent manner.Furthermore,based on our mice data,these results are transferable to other mammal species.
基金supported by the National Natural Science Foundation of China (Nos. 30700586, 31072067, 31272481)the National Spark Program Project (No. 2012GA780001)the China Scholarship Council (NO. 201308440210)
文摘Effects of antibiotic residues on methane production in anaerobic digestion are commonly studied using the following two antibiotic addition methods：（1） adding manure from animals that consume a diet containing antibiotics, and（2） adding antibiotic-free animal manure spiked with antibiotics. This study used chlortetracycline（CTC） as a model antibiotic to examine the effects of the antibiotic addition method on methane production in anaerobic digestion under two different swine wastewater concentrations（0.55 and 0.22 mg CTC/g dry manure）. The results showed that CTC degradation rate in which manure was directly added at 0.55 mg CTC/g（HSPIKE treatment） was lower than the control values and the rest of the treatment groups. Methane production from the HSPIKEtreatment was reduced（p 〈 0.05） by 12% during the whole experimental period and 15% during the first 7 days. The treatments had no significant effect on the pH and chemical oxygen demand value of the digesters, and the total nitrogen of the0.55 mg CTC/kg manure collected from mediated swine was significantly higher than the other values. Therefore, different methane production under different antibiotic addition methods might be explained by the microbial activity and the concentrations of antibiotic intermediate products and metabolites. Because the primary entry route of veterinary antibiotics into an anaerobic digester is by contaminated animal manure, the most appropriate method for studying antibiotic residue effects on methane production may be using manure from animals that are given a particular antibiotic, rather than adding the antibiotic directly to the anaerobic digester.
