Background Felids are the only definitive hosts ofToxoplasma gondii. However, the biological features of the feline small intestine followingT. gondii infection are poorly understood. We investigated the changes in th...Background Felids are the only definitive hosts ofToxoplasma gondii. However, the biological features of the feline small intestine followingT. gondii infection are poorly understood. We investigated the changes in the expression of RNAs (including mRNAs, long non-coding RNAs and circular RNAs) in the small intestinal epithelia of cats followingT. gondii infection to improve our understanding of the life cycle ofT. gondii and cat responses toT. gondii infection.Methods Fifteen cats were randomly assigned to five groups, and the infection groups were inoculated with 600 tissue cysts of theT. gondii Pru strain by gavage. The small intestinal epithelia of cats were collected at 6, 10, 14, and 30 days post infection (DPI). Using high-throughput RNA sequencing (RNA-seq), we investigated the changes in RNA expression. The expression levels of differentially expressed (DE) genes and non-coding RNAs (ncRNAs) identified by RNA-seq were validated by quantitative reverse transcription PCR (qRT-PCR). Differential expression was determined using the DESeq R package.Results In total, 207 annotated lncRNAs, 20,552 novel lncRNAs, 3342 novel circRNAs and 19,409 mRNAs were identified. Among these, 70 to 344 DE mRNAs, lncRNAs and circRNAs were detected, and the post-cleavage binding sites between 725 ncRNAs and 2082 miRNAs were predicted. Using the co-location method, we predicted that a total of 235 lncRNAs target 1044 protein-coding genes, while the results of co-expression analysis revealed that 174 lncRNAs target 2097 mRNAs. Pathway enrichment analyses of the genes targeted by ncRNAs suggested that most ncRNAs were significantly enriched in immune or diseases-related pathways. NcRNA regulatory networks revealed that a single ncRNA could be directly or indirectly regulated by multiple genes or ncRNAs that could influence the immune response of cats. Co-expression analysis showed that 242 circRNAs, mainly involved in immune responses, were significantly associated withT. gondii infection. In contrast, 1352 protein coding RNAs, mainly involved in nucleic acid process/repair pathways or oocyte development pathways, were negatively associated withT. gondii infection.Conclusions This study is the first to reveal the expression profiles of circRNAs, lncRNAs and mRNAs in the cat small intestine followingT. gondii infection and will facilitate the elucidation of the role of ncRNAs in the pathogenesis ofT. gondii infection in its definitive host, thereby facilitating the development of novel intervention strategies againstT. gondii infection in humans and animals.展开更多
AIM To investigate the effects of active vitamin D3 on autophagy and interleukin(IL)-1β expression in Salmonella-infected intestinal epithelial cells(IECs).METHODS Caco-2 cells, NOD2 siR NA-, Atg16L1 siR NA- or vitam...AIM To investigate the effects of active vitamin D3 on autophagy and interleukin(IL)-1β expression in Salmonella-infected intestinal epithelial cells(IECs).METHODS Caco-2 cells, NOD2 siR NA-, Atg16L1 siR NA- or vitamin D receptor(VDR) siR NA-transfected Caco-2 cells were pretreated with 1,25-dihydroxyvitamin D3(1,25D3), and then infected by wild-type S. typhimurium strain SL1344. The conversion of LC3-I to LC3-II was detected by Western blot analysis and LC3+ autophagosome was analyzed by immunofluorescence. Caco-2 cells or VDR si RNA-transfected cells were pretreated with 1,25D3, and then infected by SL1344. Membrane protein and total RNA were analyzed by Western blot and RT-PCR for VDR and Atg16L1 protein and m RNA expression, respectively. Atg16L1 si RNA-transfected Caco-2 cells were pretreated by 1,25D3 and then infected with SL1344. Total RNA was analyzed by RT-PCR for IL-1β mR NA expression.RESULTS The active form of vitamin D, 1,25D3, showed enhanced VDR-mediated Atg16L1 mR NA expression, membranous Atg16L1 protein expression leading to autophagic LC3 II proteins expression and LC3 punctae in Salmonella-infected Caco-2 cells which was counteracted by Atg16L1 and VDR si RNA, but Atg16L1 mediated suppression of IL-1β expression. Thus, active vitamin D may enhance autophagy but suppress inflammatory IL-1β expression in Salmonella-infected IECs.CONCLUSION Active vitamin D might enhance autophagic clearance of Salmonella infection, while modulation of inflammatory responses prevents the host from detrimental effects of overwhelming inflammation.展开更多
Heparins show great anticoagulant effect with few side effects,and are administered by subcutaneous or intravenous route in clinics.To improve patient compliance,oral administration is an alternative route.Nonetheless...Heparins show great anticoagulant effect with few side effects,and are administered by subcutaneous or intravenous route in clinics.To improve patient compliance,oral administration is an alternative route.Nonetheless,oral administration of heparins still faces enormous challenges due to the multiple obstacles.This review briefly analyzes a series of barriers ranging from poorly physicochemical properties of heparins,to harsh biological barriers including gastrointestinal degradation and pre-systemic metabolism.Moreover,several approaches have been developed to overcome these obstacles,such as improving stability of heparins in the gastrointestinal tract,enhancing the intestinal epithelia permeability and facilitating lymphatic delivery of heparins.Overall,this review aims to provide insights concerning advanced delivery strategies facilitating oral absorption of heparins.展开更多
Objective:To explore the effect of Tangshen Formula(TSF),a Chinese herbal medicine,on interstitial cells of Cajal(ICC)in the colon of diabetic rats.Methods:Fifty-four male Wistar rats were randomly divided into normal...Objective:To explore the effect of Tangshen Formula(TSF),a Chinese herbal medicine,on interstitial cells of Cajal(ICC)in the colon of diabetic rats.Methods:Fifty-four male Wistar rats were randomly divided into normal control(NC,n=14)and high-fat diet(HFD)groups(n=40).After 6 weeks,the rats in the HFD group were injected intraperitoneally streptozotocin once(30 mg/kg).Thirty rats with fasting blood glucose higher than 11.7 mmol/L were randomly divided into diabetes(DM)and TSF groups,15 rats in each group.Rats in the NC and DM groups were intragastrically administered with saline,and those in the TSF group were given with TSF(2.4 g/kg)once daily for 20 weeks.Expression levels of Bax,Bcl-2,and caspase-3 in colonic smooth muscle layer were measured by Western blotting and immunohistochemical staining.The number of ICC was determined by immunohistochemical staining.Immunofluorescence was used for analyzing the ratio of classically activated macrophages(M1)and alternatively activated macrophages(M2)to total macrophages.Electron microscopy was used to observe the epithelial ultrastructure and junctions.Results:TSF appeared to partially prevented loss of ICC in DM rats(P<0.05).Compared with the NC group,expression levels of Bcl-2,Bax,caspase-3,and TNF-αas well as the ratio of M1 to total macrophages increased in DM rats(all P<0.05),and the ratio of M2 to total macrophages decreased(P<0.05 or P<0.01).Compared with the DM group,TSF decreased the expression levels of abovementioned proteins and restore M2 to total macrophages ratio(P<0.05 or P<0.01).TSF appeared to attenuate the ultrastructural changes of epithelia and improve the tight and desmosome junctions between epithelia reduced in the DM rats.Conclusions:Reduced number of ICC in DM rats may be associated with damage of the intestinal barrier.The protective effects of TSF on ICC may be through repair of the epithelial junctions,which attenuates inflammation and inflammation-initiated apoptosis in colon of DM rats.展开更多
基金Project support was kindly provided by the National Key Research and Development Program of China(Grant Nos.2021YFC2300800 and 2021YFC2300802)the National Natural Science Foundation of China(Grant Nos.32172887 and 32102701)+4 种基金the Youth Science and Technology Fund Program of Gansu province(Grant No.23JRRA562)the Innovation Project of Chinese Academy of Agricultural Sciences(Grant No.25-LZIHPS-05)the Agricultural Science and Technology Innovation Program(ASTIP)(Grant No.CAAS-ASTIP-2016-LVRI-03)the Yunnan Expert Workstation(Grant No.202005AF150041)The funding bodies played no role in the design of the study and collection,analysis,and interpretation of data and in writing the manuscript.
文摘Background Felids are the only definitive hosts ofToxoplasma gondii. However, the biological features of the feline small intestine followingT. gondii infection are poorly understood. We investigated the changes in the expression of RNAs (including mRNAs, long non-coding RNAs and circular RNAs) in the small intestinal epithelia of cats followingT. gondii infection to improve our understanding of the life cycle ofT. gondii and cat responses toT. gondii infection.Methods Fifteen cats were randomly assigned to five groups, and the infection groups were inoculated with 600 tissue cysts of theT. gondii Pru strain by gavage. The small intestinal epithelia of cats were collected at 6, 10, 14, and 30 days post infection (DPI). Using high-throughput RNA sequencing (RNA-seq), we investigated the changes in RNA expression. The expression levels of differentially expressed (DE) genes and non-coding RNAs (ncRNAs) identified by RNA-seq were validated by quantitative reverse transcription PCR (qRT-PCR). Differential expression was determined using the DESeq R package.Results In total, 207 annotated lncRNAs, 20,552 novel lncRNAs, 3342 novel circRNAs and 19,409 mRNAs were identified. Among these, 70 to 344 DE mRNAs, lncRNAs and circRNAs were detected, and the post-cleavage binding sites between 725 ncRNAs and 2082 miRNAs were predicted. Using the co-location method, we predicted that a total of 235 lncRNAs target 1044 protein-coding genes, while the results of co-expression analysis revealed that 174 lncRNAs target 2097 mRNAs. Pathway enrichment analyses of the genes targeted by ncRNAs suggested that most ncRNAs were significantly enriched in immune or diseases-related pathways. NcRNA regulatory networks revealed that a single ncRNA could be directly or indirectly regulated by multiple genes or ncRNAs that could influence the immune response of cats. Co-expression analysis showed that 242 circRNAs, mainly involved in immune responses, were significantly associated withT. gondii infection. In contrast, 1352 protein coding RNAs, mainly involved in nucleic acid process/repair pathways or oocyte development pathways, were negatively associated withT. gondii infection.Conclusions This study is the first to reveal the expression profiles of circRNAs, lncRNAs and mRNAs in the cat small intestine followingT. gondii infection and will facilitate the elucidation of the role of ncRNAs in the pathogenesis ofT. gondii infection in its definitive host, thereby facilitating the development of novel intervention strategies againstT. gondii infection in humans and animals.
基金Supported by the Ministry of Science and Technology[MOST 103-2314-B-182-032(in part)]the Chang Gung Memorial Hospital,No.CMRPG8B1431,No.CMRPG8B1481 and No.CMRPG880443the Stem Cell Research Core Laboratory (grant CLRPG8B0052) for technical support
文摘AIM To investigate the effects of active vitamin D3 on autophagy and interleukin(IL)-1β expression in Salmonella-infected intestinal epithelial cells(IECs).METHODS Caco-2 cells, NOD2 siR NA-, Atg16L1 siR NA- or vitamin D receptor(VDR) siR NA-transfected Caco-2 cells were pretreated with 1,25-dihydroxyvitamin D3(1,25D3), and then infected by wild-type S. typhimurium strain SL1344. The conversion of LC3-I to LC3-II was detected by Western blot analysis and LC3+ autophagosome was analyzed by immunofluorescence. Caco-2 cells or VDR si RNA-transfected cells were pretreated with 1,25D3, and then infected by SL1344. Membrane protein and total RNA were analyzed by Western blot and RT-PCR for VDR and Atg16L1 protein and m RNA expression, respectively. Atg16L1 si RNA-transfected Caco-2 cells were pretreated by 1,25D3 and then infected with SL1344. Total RNA was analyzed by RT-PCR for IL-1β mR NA expression.RESULTS The active form of vitamin D, 1,25D3, showed enhanced VDR-mediated Atg16L1 mR NA expression, membranous Atg16L1 protein expression leading to autophagic LC3 II proteins expression and LC3 punctae in Salmonella-infected Caco-2 cells which was counteracted by Atg16L1 and VDR si RNA, but Atg16L1 mediated suppression of IL-1β expression. Thus, active vitamin D may enhance autophagy but suppress inflammatory IL-1β expression in Salmonella-infected IECs.CONCLUSION Active vitamin D might enhance autophagic clearance of Salmonella infection, while modulation of inflammatory responses prevents the host from detrimental effects of overwhelming inflammation.
基金Supported by the Natural Science Fund for Colleges and Universities in Jiangsu Province(No.18KJB350009)the Natural Science Fund for Colleges and Universities in Jiangsu Province(No.17KJB350009)the Natural Science Foundation of Jiangsu Province(No.BK20170445).
文摘Heparins show great anticoagulant effect with few side effects,and are administered by subcutaneous or intravenous route in clinics.To improve patient compliance,oral administration is an alternative route.Nonetheless,oral administration of heparins still faces enormous challenges due to the multiple obstacles.This review briefly analyzes a series of barriers ranging from poorly physicochemical properties of heparins,to harsh biological barriers including gastrointestinal degradation and pre-systemic metabolism.Moreover,several approaches have been developed to overcome these obstacles,such as improving stability of heparins in the gastrointestinal tract,enhancing the intestinal epithelia permeability and facilitating lymphatic delivery of heparins.Overall,this review aims to provide insights concerning advanced delivery strategies facilitating oral absorption of heparins.
基金Supported by National Natural Science Foundation of China(No.81873135)。
文摘Objective:To explore the effect of Tangshen Formula(TSF),a Chinese herbal medicine,on interstitial cells of Cajal(ICC)in the colon of diabetic rats.Methods:Fifty-four male Wistar rats were randomly divided into normal control(NC,n=14)and high-fat diet(HFD)groups(n=40).After 6 weeks,the rats in the HFD group were injected intraperitoneally streptozotocin once(30 mg/kg).Thirty rats with fasting blood glucose higher than 11.7 mmol/L were randomly divided into diabetes(DM)and TSF groups,15 rats in each group.Rats in the NC and DM groups were intragastrically administered with saline,and those in the TSF group were given with TSF(2.4 g/kg)once daily for 20 weeks.Expression levels of Bax,Bcl-2,and caspase-3 in colonic smooth muscle layer were measured by Western blotting and immunohistochemical staining.The number of ICC was determined by immunohistochemical staining.Immunofluorescence was used for analyzing the ratio of classically activated macrophages(M1)and alternatively activated macrophages(M2)to total macrophages.Electron microscopy was used to observe the epithelial ultrastructure and junctions.Results:TSF appeared to partially prevented loss of ICC in DM rats(P<0.05).Compared with the NC group,expression levels of Bcl-2,Bax,caspase-3,and TNF-αas well as the ratio of M1 to total macrophages increased in DM rats(all P<0.05),and the ratio of M2 to total macrophages decreased(P<0.05 or P<0.01).Compared with the DM group,TSF decreased the expression levels of abovementioned proteins and restore M2 to total macrophages ratio(P<0.05 or P<0.01).TSF appeared to attenuate the ultrastructural changes of epithelia and improve the tight and desmosome junctions between epithelia reduced in the DM rats.Conclusions:Reduced number of ICC in DM rats may be associated with damage of the intestinal barrier.The protective effects of TSF on ICC may be through repair of the epithelial junctions,which attenuates inflammation and inflammation-initiated apoptosis in colon of DM rats.
