[Objective] The aim was to study the polymorphism of CMYA3 gene in the 148 pigs of hybrid offspring of 13/17 Robertsonian translocation pigs [2n = 37,rob （13;17）] intercrossing.[Method] PCR-RFLP method was adopted.[...[Objective] The aim was to study the polymorphism of CMYA3 gene in the 148 pigs of hybrid offspring of 13/17 Robertsonian translocation pigs [2n = 37,rob （13;17）] intercrossing.[Method] PCR-RFLP method was adopted.[Result] A 507 bp fragment of CMYA3 gene was obtained by PCR amplification,and then amplification product by using restriction nuclease Bsh1236Ⅰ was detected by agarose gel electrophoresis.As a result,both alleles （A and B） of the loci were found in the population.The frequencies of allele A and B were 0.699 and 0.301.The genotype frequencies of AA,AB and BB were 0.615,0.169 and 0.216.The frequencies of allele A and genotype AA were significantly higher than allele B and genotype BB in populations.[Conclusion] The study will provide theoretical basis for molecular breeding and marker-assisted selection of 13/17 Robertsonian translocation pigs.展开更多
AIM: To investigate into the potential involvement of pyrin containing 3 gene(NLRP3), a member of the nucleotide-binding oligomerization domain-like receptors with cytosolic pattern recognition, in the host defense of...AIM: To investigate into the potential involvement of pyrin containing 3 gene(NLRP3), a member of the nucleotide-binding oligomerization domain-like receptors with cytosolic pattern recognition, in the host defense of corneas against viruses.METHODS: The herpes viral keratitis model was utilized in BALB/c mice with inoculation of herpes simplex virus-1(HSV-1). Corneal tissues removed during therapy of patients with viral keratitis as well as a Simian vacuolating virus 40(SV40)-immortalized human corneal epithelial cell line were also examined.Immunohistochemistry was used to detect NLRP3 in these subjects, focusing on their distribution in tissue or cells. Western blot was used to measure the level of NLRP3 and another two related molecules in NLPR3 inflammasome, namely caspase-1 and IL-1β.RESULTS: The NLRP3 activation induced by HSV-1infection in corneas was accompanied with redistribution of NLRP3 from the cytoplasm to the nucleus in both murine and human corneal epithelial cells. Furthermore,in the SV40-immortalized human corneal epithelial cells,NLRP3 was exclusively located in the nucleus, and treatment of the cells with high concentration of extracellular potassium(known as an inhibitor of NLRP3activation) effectively drove NLRP3 back to the cytoplasm as reflected by both immunohistochemistry and Western blot.· CONCLUSION: It is proposed that herpes virus infection activates and causes redistribution of NLRP3 to nuclei. Whether this NLRP3 translocation occurs with other viral infections and in other cell types merit further study.展开更多
BACKGROUND: It has been found in recent years that STAT3 widely distributes in nervous system, including hippocampal CA1-3 region, dentate gyrus and cerebral neocortex, etc. Ischemic brain injury can cause the release...BACKGROUND: It has been found in recent years that STAT3 widely distributes in nervous system, including hippocampal CA1-3 region, dentate gyrus and cerebral neocortex, etc. Ischemic brain injury can cause the release of some cytokines and growth factors, while electro-acupuncture may have multi-level, multi-channel and multi-target protective and interventional effects on ischemic brain injury. OBJECTIVE: To observe the effects of electro-acupuncture on STAT3 expression and nuclear translocation in hippocampal CA1 region of rat models of brain ischemia/reperfusion. DESIGN: Randomized and controlled observation. SETTING: Staff Room of Acupuncture and Moxibustion, Department of Acupuncture and Bone Injury, Hubei College of Traditional Chinese Medicine; Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Seventy-two healthy SD rats, of clean degree and either gender, weighing (200±20) g, were provided by the Experimental Animal Center of Hubei College of Traditional Chinese Medicine. STAT3 monoclonal antibody was purchased from Santa Cruz Company, USA, and G-6805 electro-acupuncture instrument was purchased from Shanghai Medical Electronic Instruments Factory. METHODS: This experiment was carried out in the comprehensive laboratory of Department of Acupuncture and Bone Injury, Hubei College of Traditional Chinese Medicine between September 2005 and February 2006. Seventy-two rats were randomly divided into 4 groups: ① control group(n =6): Untouched. ② Sham-operation group (n =18): Artery was isolated, but without inserting thread bolt.③ Model group (n =24): Rat models of local brain ischemia/reperfusion were established with modified suture occlusion. ④Electro-acupuncture group (n =24): Dazhui and bilateral Neiguan points were selected for electro-acupuncture treatment. No. 28 acupuncture needle of 3.33 cm was used in the treatment. A G-6085 electro-acupuncture instrument with continuous wave, frequency of 120 times/min, intensity of 1 mA, 30 min/time, was used. Acupuncture was conducted firstly at ischemia/reperfusion 3 hours, then once every 12 hours. STAT3 positive nuclear translocation in hippocampal CA1 region of rats was observed with immunohistochemical method at 24, 48 and 72 hours after brain ishcemia/reperfusion, and then STAT3 positive cells were counted. MAIN OUTCOME MEASURES: STAT3 positive cells and nuclear translocation in hippocampal CA1 region of rats in each group. RESULTS: All the 72 rats were involved in the result analysis. ①In the control group and sham-operation group, STAT3 positive cells with light cytoplasm and nucleus were decreased , and nuclear translocation was not found. ② In the model group, STAT3 positive cells were mostly found in the cytoplasm of the hippocampal CA1 region at the ischemic side of rats after ischemia/reperfusion 24 hours. They were significantly more than those in the sham-operation group and control group [(18.00±2.68), (9.00±1.35), (8.00±1.22) cells/ mm2, P < 0.01], but cells with nuclear reaction were fewer; At ischemia/reperfusion 48 and 72 hours, STAT3 positive cells were increased, and they were significantly more than those of sham-operation group [(25.00±3.23), (35.00±3.52) cells/mm2, (13.00±1.93), (12.00±1.24) cells/mm2, P < 0.01]. Positive cells with nuclear reaction were found dark-stained. ③At ischemia/reperfusion 24, 48 and 72 hours, STAT3 positive cells were strongly expressed in hippocampal CA1 region at ischemic side of rats of electro-acupuncture group, and they were significantly more than those of model group [(25±3.52), (50±6.31), (75±8.09) cells/mm2, P < 0.01]. STAT3 positive cells were gradually enhanced with time, and considerable STAT3 nuclear positive reaction cells were found. CONCLUSION: Electro-acupuncture can activate STAT3 protein expression in hippocampal tissue of rats with local brain ischemia/reperfusion, promote STAT3 nuclear translocation and function its neuroprotective effect.展开更多
Gene fusions involving two of the MiT subfamily factors, such as TFE3, TFEB, TFC and MiTF, have been identified in renal cell carcinoma (RCC). Xp11.2 translocation RCC is a rare pediatric neoplasm that harbors gene fu...Gene fusions involving two of the MiT subfamily factors, such as TFE3, TFEB, TFC and MiTF, have been identified in renal cell carcinoma (RCC). Xp11.2 translocation RCC is a rare pediatric neoplasm that harbors gene fusions involving TFE3, which plays an important role in cell proliferation and survival. We herein present a case of RCC associated with Xp11.2 translocation/TFE3 gene fusion in a 14-year-old Japanese boy presenting gross hematuria and body weight loss. The tumor was characterized by histopathology, cytology and TFE3-immunohistochemistry/immunocytochemistry. Knowledge of distinctive morphological and immunostaining features of this tumor can help to accurately diagnose this rare subset of translocation associated RCC in routine pathological diagnostic procedures.展开更多
The recognition of cytosolic nucleic acid triggers the DNA/RNA sensor–IRF3 axis-mediated production of type I interferons(IFNs),which are essential for antiviral immune responses.However,the inappropriate activation ...The recognition of cytosolic nucleic acid triggers the DNA/RNA sensor–IRF3 axis-mediated production of type I interferons(IFNs),which are essential for antiviral immune responses.However,the inappropriate activation of these signaling pathways is implicated in autoimmune conditions.Here,we report that indomethacin,a widely used nonsteroidal anti-inflammatory drug,inhibits nucleic acid-triggered IFN production.We found that both DNA-and RNA-stimulated IFN expression can be effectively blocked by indomethacin.Interestingly,indomethacin also prohibits the nuclear translocation of IRF3 following cytosolic nucleic acid recognition.Importantly,in cell lines and a mouse model of Aicardi–Goutières syndrome,indomethacin administration blunts self-DNA-induced autoimmune responses.Thus,our study reveals a previously unknown function of indomethacin and provides a potential treatment for cytosolic nucleic acid-stimulated autoimmunity.展开更多
Objective To investigate the potential therapeutic targets and pharmacological mechanism of(-)-epigallocatechin-3-gallate(EGCG)based on network pharmacology and experimental verification.METHODS The druggability of EG...Objective To investigate the potential therapeutic targets and pharmacological mechanism of(-)-epigallocatechin-3-gallate(EGCG)based on network pharmacology and experimental verification.METHODS The druggability of EGCG was measured by the traditional Chinese medicine systems pharmacology(TCMSP)server,and potential targets of EGCG were identified by Pharm Mapper and Drug Repositioning and Adverse drug Reaction via Chemical-Protein Interactome(DRAR-CPI).The potential targets were imported into GeneMANIA database to obtain the protein-protein direct interaction network,and target physical interaction,co-expression,prediction,genetic interaction,and shared protein domains.The biological process,molecular functions,cellular components and KEGG signaling pathways of potential targets were analyzed using DAVID database.For further study,ethanol was used to establish a model of endothelial injury in vitro.The cell viability was assayed by MTT method,the cellular apoptosis was stained by Annexin V/PI,and the expression levels of Bcl-2,Bax and cleved-caspase-3 were tested by Western blotting.Then,JC-1 and nuclear translocation of NF-κB experiments were used to study the mitochondrial membrane potential and nuclear translocation.RESULTS The oral availability of EGCG was 55.09%(≥30%)and drug-like index was 0.77(≥0.18),which were considered pharmacokinetically active.17 potential targetable proteins of EGCG were predicted by Pharm Mapper and DRAR-CPI.Further research showed that 68.13%displayed similar co-expression characteristics,26.11%physical interactions,and 2.74%shared the same protein domain.The depth network analysis results showed that the biofunctions of EGCG were mainly by regulating glutathione derivative biosynthetic process,glutathione metabolic process,nitrogen compound metabolic process etc..via drug binding,catalytic activity,glutathione transferase activity,anion binding etc..in sarcoplasmic reticulum,spindle pole,microtubule cytoskeleton and cytoplasm.KEGG enrichment analysis showed that Glutathione metabolism,IL^(-1)7 signaling pathway,EGFR tyrosine kinase inhibitor resistance,PI3K-Akt signaling pathway and other pathways were involves in the biofunction of EGCG.The above analyses indicated that EGCG exerts its biofunction through antioxidant and anti-inflammatory mechanisms.The experimental results showed that ethanol 20.0 mmol·L^(-1) decreased cell viability,Bcl-2 expression,and increased cell apoptosis,the intracellular ROS,as well as the expression of Bax and cleaved-caspase-3 of human endothelial cells.However,treatment of the cells with EGCG can significantly alleviate ethanol induced endothelial cells injury.Further study showed that EGCG significantly alleviates ethanol induced mitochondrial depolarization and nuclear translocation of NF-κB.CONCLUSIONS EGCG exerts pharmacological efficacies on ethanol induced endothelial cell injury through multi-target,multi-function and multi-pathway mode.Protective effect of EGCG on ethanol induced cell injury was mainly through alteration of mitochondrial function and NF-κB translocation.Therefore,EGCG have great potential in protecting against endothelial dysfunction of the persons who are chronically abuse of ethanol.This study also provides a new understanding of EGCG in clinical application on cardiovascular and cerebrovascular diseases.展开更多
基金Supported by Major Specialized Subject of Transgenic Organism New Variety Breeding(20082X08006-003)National Natural Science Foundation of China(30871778 )Construction Engineering Special Fund for Mountain Tai Scholars of Shandong Province~~
文摘[Objective] The aim was to study the polymorphism of CMYA3 gene in the 148 pigs of hybrid offspring of 13/17 Robertsonian translocation pigs [2n = 37,rob （13;17）] intercrossing.[Method] PCR-RFLP method was adopted.[Result] A 507 bp fragment of CMYA3 gene was obtained by PCR amplification,and then amplification product by using restriction nuclease Bsh1236Ⅰ was detected by agarose gel electrophoresis.As a result,both alleles （A and B） of the loci were found in the population.The frequencies of allele A and B were 0.699 and 0.301.The genotype frequencies of AA,AB and BB were 0.615,0.169 and 0.216.The frequencies of allele A and genotype AA were significantly higher than allele B and genotype BB in populations.[Conclusion] The study will provide theoretical basis for molecular breeding and marker-assisted selection of 13/17 Robertsonian translocation pigs.
基金Supported by National Natural Science Foundation of China(No.81273212,81100651)Project of Science and Technology of Shandong Province(No.2014GSF118044)
文摘AIM: To investigate into the potential involvement of pyrin containing 3 gene(NLRP3), a member of the nucleotide-binding oligomerization domain-like receptors with cytosolic pattern recognition, in the host defense of corneas against viruses.METHODS: The herpes viral keratitis model was utilized in BALB/c mice with inoculation of herpes simplex virus-1(HSV-1). Corneal tissues removed during therapy of patients with viral keratitis as well as a Simian vacuolating virus 40(SV40)-immortalized human corneal epithelial cell line were also examined.Immunohistochemistry was used to detect NLRP3 in these subjects, focusing on their distribution in tissue or cells. Western blot was used to measure the level of NLRP3 and another two related molecules in NLPR3 inflammasome, namely caspase-1 and IL-1β.RESULTS: The NLRP3 activation induced by HSV-1infection in corneas was accompanied with redistribution of NLRP3 from the cytoplasm to the nucleus in both murine and human corneal epithelial cells. Furthermore,in the SV40-immortalized human corneal epithelial cells,NLRP3 was exclusively located in the nucleus, and treatment of the cells with high concentration of extracellular potassium(known as an inhibitor of NLRP3activation) effectively drove NLRP3 back to the cytoplasm as reflected by both immunohistochemistry and Western blot.· CONCLUSION: It is proposed that herpes virus infection activates and causes redistribution of NLRP3 to nuclei. Whether this NLRP3 translocation occurs with other viral infections and in other cell types merit further study.
基金the Natural Science Foundation of Hubei Province,No.2003ABA154
文摘BACKGROUND: It has been found in recent years that STAT3 widely distributes in nervous system, including hippocampal CA1-3 region, dentate gyrus and cerebral neocortex, etc. Ischemic brain injury can cause the release of some cytokines and growth factors, while electro-acupuncture may have multi-level, multi-channel and multi-target protective and interventional effects on ischemic brain injury. OBJECTIVE: To observe the effects of electro-acupuncture on STAT3 expression and nuclear translocation in hippocampal CA1 region of rat models of brain ischemia/reperfusion. DESIGN: Randomized and controlled observation. SETTING: Staff Room of Acupuncture and Moxibustion, Department of Acupuncture and Bone Injury, Hubei College of Traditional Chinese Medicine; Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Seventy-two healthy SD rats, of clean degree and either gender, weighing (200±20) g, were provided by the Experimental Animal Center of Hubei College of Traditional Chinese Medicine. STAT3 monoclonal antibody was purchased from Santa Cruz Company, USA, and G-6805 electro-acupuncture instrument was purchased from Shanghai Medical Electronic Instruments Factory. METHODS: This experiment was carried out in the comprehensive laboratory of Department of Acupuncture and Bone Injury, Hubei College of Traditional Chinese Medicine between September 2005 and February 2006. Seventy-two rats were randomly divided into 4 groups: ① control group(n =6): Untouched. ② Sham-operation group (n =18): Artery was isolated, but without inserting thread bolt.③ Model group (n =24): Rat models of local brain ischemia/reperfusion were established with modified suture occlusion. ④Electro-acupuncture group (n =24): Dazhui and bilateral Neiguan points were selected for electro-acupuncture treatment. No. 28 acupuncture needle of 3.33 cm was used in the treatment. A G-6085 electro-acupuncture instrument with continuous wave, frequency of 120 times/min, intensity of 1 mA, 30 min/time, was used. Acupuncture was conducted firstly at ischemia/reperfusion 3 hours, then once every 12 hours. STAT3 positive nuclear translocation in hippocampal CA1 region of rats was observed with immunohistochemical method at 24, 48 and 72 hours after brain ishcemia/reperfusion, and then STAT3 positive cells were counted. MAIN OUTCOME MEASURES: STAT3 positive cells and nuclear translocation in hippocampal CA1 region of rats in each group. RESULTS: All the 72 rats were involved in the result analysis. ①In the control group and sham-operation group, STAT3 positive cells with light cytoplasm and nucleus were decreased , and nuclear translocation was not found. ② In the model group, STAT3 positive cells were mostly found in the cytoplasm of the hippocampal CA1 region at the ischemic side of rats after ischemia/reperfusion 24 hours. They were significantly more than those in the sham-operation group and control group [(18.00±2.68), (9.00±1.35), (8.00±1.22) cells/ mm2, P < 0.01], but cells with nuclear reaction were fewer; At ischemia/reperfusion 48 and 72 hours, STAT3 positive cells were increased, and they were significantly more than those of sham-operation group [(25.00±3.23), (35.00±3.52) cells/mm2, (13.00±1.93), (12.00±1.24) cells/mm2, P < 0.01]. Positive cells with nuclear reaction were found dark-stained. ③At ischemia/reperfusion 24, 48 and 72 hours, STAT3 positive cells were strongly expressed in hippocampal CA1 region at ischemic side of rats of electro-acupuncture group, and they were significantly more than those of model group [(25±3.52), (50±6.31), (75±8.09) cells/mm2, P < 0.01]. STAT3 positive cells were gradually enhanced with time, and considerable STAT3 nuclear positive reaction cells were found. CONCLUSION: Electro-acupuncture can activate STAT3 protein expression in hippocampal tissue of rats with local brain ischemia/reperfusion, promote STAT3 nuclear translocation and function its neuroprotective effect.
文摘Gene fusions involving two of the MiT subfamily factors, such as TFE3, TFEB, TFC and MiTF, have been identified in renal cell carcinoma (RCC). Xp11.2 translocation RCC is a rare pediatric neoplasm that harbors gene fusions involving TFE3, which plays an important role in cell proliferation and survival. We herein present a case of RCC associated with Xp11.2 translocation/TFE3 gene fusion in a 14-year-old Japanese boy presenting gross hematuria and body weight loss. The tumor was characterized by histopathology, cytology and TFE3-immunohistochemistry/immunocytochemistry. Knowledge of distinctive morphological and immunostaining features of this tumor can help to accurately diagnose this rare subset of translocation associated RCC in routine pathological diagnostic procedures.
基金supported by grants from the National Natural Science Foundation of China(82001690 to H.C.81925017 and 82130052 to T.L.)the National Key Research and Development Program of China(2020YFA0707703 to T.Z.and 2020YFA0707702 to T.L.).
文摘The recognition of cytosolic nucleic acid triggers the DNA/RNA sensor–IRF3 axis-mediated production of type I interferons(IFNs),which are essential for antiviral immune responses.However,the inappropriate activation of these signaling pathways is implicated in autoimmune conditions.Here,we report that indomethacin,a widely used nonsteroidal anti-inflammatory drug,inhibits nucleic acid-triggered IFN production.We found that both DNA-and RNA-stimulated IFN expression can be effectively blocked by indomethacin.Interestingly,indomethacin also prohibits the nuclear translocation of IRF3 following cytosolic nucleic acid recognition.Importantly,in cell lines and a mouse model of Aicardi–Goutières syndrome,indomethacin administration blunts self-DNA-induced autoimmune responses.Thus,our study reveals a previously unknown function of indomethacin and provides a potential treatment for cytosolic nucleic acid-stimulated autoimmunity.
基金National Natural Science Foundation of China(82100488)Key Research and Development Pro⁃gram Project of Shaanxi Province(2021SF-071)and National Training Program of Innovation and Entrepreneurship for Students of China(201910716019,201910716020,202110716027)。
文摘Objective To investigate the potential therapeutic targets and pharmacological mechanism of(-)-epigallocatechin-3-gallate(EGCG)based on network pharmacology and experimental verification.METHODS The druggability of EGCG was measured by the traditional Chinese medicine systems pharmacology(TCMSP)server,and potential targets of EGCG were identified by Pharm Mapper and Drug Repositioning and Adverse drug Reaction via Chemical-Protein Interactome(DRAR-CPI).The potential targets were imported into GeneMANIA database to obtain the protein-protein direct interaction network,and target physical interaction,co-expression,prediction,genetic interaction,and shared protein domains.The biological process,molecular functions,cellular components and KEGG signaling pathways of potential targets were analyzed using DAVID database.For further study,ethanol was used to establish a model of endothelial injury in vitro.The cell viability was assayed by MTT method,the cellular apoptosis was stained by Annexin V/PI,and the expression levels of Bcl-2,Bax and cleved-caspase-3 were tested by Western blotting.Then,JC-1 and nuclear translocation of NF-κB experiments were used to study the mitochondrial membrane potential and nuclear translocation.RESULTS The oral availability of EGCG was 55.09%(≥30%)and drug-like index was 0.77(≥0.18),which were considered pharmacokinetically active.17 potential targetable proteins of EGCG were predicted by Pharm Mapper and DRAR-CPI.Further research showed that 68.13%displayed similar co-expression characteristics,26.11%physical interactions,and 2.74%shared the same protein domain.The depth network analysis results showed that the biofunctions of EGCG were mainly by regulating glutathione derivative biosynthetic process,glutathione metabolic process,nitrogen compound metabolic process etc..via drug binding,catalytic activity,glutathione transferase activity,anion binding etc..in sarcoplasmic reticulum,spindle pole,microtubule cytoskeleton and cytoplasm.KEGG enrichment analysis showed that Glutathione metabolism,IL^(-1)7 signaling pathway,EGFR tyrosine kinase inhibitor resistance,PI3K-Akt signaling pathway and other pathways were involves in the biofunction of EGCG.The above analyses indicated that EGCG exerts its biofunction through antioxidant and anti-inflammatory mechanisms.The experimental results showed that ethanol 20.0 mmol·L^(-1) decreased cell viability,Bcl-2 expression,and increased cell apoptosis,the intracellular ROS,as well as the expression of Bax and cleaved-caspase-3 of human endothelial cells.However,treatment of the cells with EGCG can significantly alleviate ethanol induced endothelial cells injury.Further study showed that EGCG significantly alleviates ethanol induced mitochondrial depolarization and nuclear translocation of NF-κB.CONCLUSIONS EGCG exerts pharmacological efficacies on ethanol induced endothelial cell injury through multi-target,multi-function and multi-pathway mode.Protective effect of EGCG on ethanol induced cell injury was mainly through alteration of mitochondrial function and NF-κB translocation.Therefore,EGCG have great potential in protecting against endothelial dysfunction of the persons who are chronically abuse of ethanol.This study also provides a new understanding of EGCG in clinical application on cardiovascular and cerebrovascular diseases.
