Transient receptor potential melastatin 2(TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression...Transient receptor potential melastatin 2(TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma(PC12) cells injured by oxygen-glucose deprivation(OGD). PC12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reoxygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2(CXCL2), NACHT, LRR, and PYD domain–containing protein 3(NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation.展开更多
Background:In the livestock industry,intramuscular fat content is a key factor affecting meat quality.Many studies have shown that dietary calcium supplementation is closely related to lipid metabolism.However,few stu...Background:In the livestock industry,intramuscular fat content is a key factor affecting meat quality.Many studies have shown that dietary calcium supplementation is closely related to lipid metabolism.However,few studies have examined the relationship between dietary calcium supplementation and intramuscular fat accumulation.Methods:Here,we used C2C12 cells,C57BL/6 mice(n=8)and three-way cross-breeding pigs(Duroc×Landrace×Large white)(n=10)to study the effect of calcium addition on intramuscular fat accumulation.In vitro,we used calcium chloride to adjust the calcium levels in the medium(2 mmol/L or 3 mmol/L).Then we measured various indicators.In vivo,calcium carbonate was used to regulate calcium levels in feeds(Mice:0.5%calcium or 1.2%calcium)(Pigs:0.9%calcium or 1.5%calcium).Then we tested the mice gastrocnemius muscle triglyceride content,pig longissimus dorsi muscle meat quality and lipidomics.Results:In vitro,calcium addition(3 mmol/L)had no significant effect on cell proliferation,but promoted the differentiation of C2C12 cells into slow-twitch fibers.Calcium supplementation increased triglyceride accumulation in C2C12 cells.Calcium addition increased the number of mitochondria and also increased the calcium level in the mitochondria and reduced the of key enzymes activity involved inβ-oxidation such as acyl-coenzyme A dehydrogenase.Decreasing mitochondrial calcium level can alleviate lipid accumulation induced by calcium addition.In addition,calcium addition also reduced the glycolytic capacity and glycolytic conversion rate of C2C12 cells.In vivo,dietary calcium supplementation(1.2%)promoted the accumulation of triglycerides in the gastrocnemius muscle of mice.Dietary calcium supplementation(1.5%)had no effect on pig weight,but significantly improved the flesh color of the longissimus dorsi muscle,reduced the backfat thickness and increased intramuscular fat content in pigs.Besides,calcium addition had no effect on longissimus dorsi pH,electrical conductivity and shear force.Conclusions:These results suggest that calcium addition promotes intramuscular fat accumulation by inhibiting the oxidation of fatty acids.These findings provide a new tool for increasing intramuscular fat content and an economical strategy for improving meat quality.展开更多
The alloy AZ91 containing calcium was prepared under protection of a mixed gas atmosphere of SF6 （1%, volume fraction） and CO2. The added calcium mainly dissolves into the Mg17Al12 phase in the alloy and increases i...The alloy AZ91 containing calcium was prepared under protection of a mixed gas atmosphere of SF6 （1%, volume fraction） and CO2. The added calcium mainly dissolves into the Mg17Al12 phase in the alloy and increases its melting point and the thermal stability. The empirical electron theory （EET） of solid and molecules was used to calculate the valence electron structures （VES） of Mg17Al12 phase with different amounts of calcium additions. The theoretical calculations indicate that calcium dissolved in Mg17Al12 phase increases the strengths of atomic bonds that control the thermal stability of Mg17Al12 phase, and also makes the distribution of valence electrons on the dominant bond network as well as in the whole unit cell of Mg17Al12 more uniform, which are consistent with the experimental results.展开更多
A series of diatomite supported Cu/Ni bimetallic catalysts were prepared using the co-impregnation method to improve the efficiency and selectivity toward methyl 12-hydroxystearate in the hydrogenation of methyl ricin...A series of diatomite supported Cu/Ni bimetallic catalysts were prepared using the co-impregnation method to improve the efficiency and selectivity toward methyl 12-hydroxystearate in the hydrogenation of methyl ricinoleate.The catalysts were characterized using X-ray diffraction(XRD),transmission electron microscopy(TEM),scanning electron microscopy and energy dispersive X-ray spectroscopy(SEM-EDS),X-ray photoelectron spectroscopy(XPS)and temperature programmed reduction(H2-TPR).All the characterization results verified the formation of highly dispersed Cu/Ni alloy on support.Moreover,by subtly regulating the Ni/Cu molar ratio as well as the reaction parameters,the hydrogenation of methyl ricinoleate to methyl 12-hydroxystearate proceeded efficiently and selectively,affording 97%yield of methyl 12-hydroxystearate and nearly equivalent conversion of methyl ricinoleate under 2 MPa H2 pressure and at 130 C in 4 h with only 1 wt%of the catalyst Ni7Cu1/diatomite(based on methyl ricinoleate).Besides,the supported Cu–Ni bimetallic catalyst is stable during recycle and reuse.After five cycles of reuse,much catalytic activity is still preserved.Therefore,this low-cost and stable bimetallic catalyst would be promising for the hydrogenation of methyl ricinoleate to methyl 12-hydroxystearate,representing an example of green catalysis for efficiently conversion of biomass to value-added chemicals and materials.展开更多
Chronic atrophic autoimmune gastritis (CAAG) is an organ-specific autoimmune disease characterized by an immune response, which is directed towards the parietal cells and intrinsic factor of the gastric body and fundu...Chronic atrophic autoimmune gastritis (CAAG) is an organ-specific autoimmune disease characterized by an immune response, which is directed towards the parietal cells and intrinsic factor of the gastric body and fundus and leads to hypochlorhydria, hypergastrinemia and inadequate production of the intrinsic factor. As a result, the stomach’s secretion of essential substances, such as hydrochloric acid and intrinsic factor, is reduced, leading to digestive impairments. The most common is vitamin B12 deficiency, which results in a megaloblastic anemia and iron malabsorption, leading to iron deficiency anemia. However, in the last years the deficiency of several other vitamins and micronutrients, such as vitamin C, vitamin D, folic acid and calcium, has been increasingly described in patients with CAAG. In addition the occurrence of multiple vitamin deficiencies may lead to severe hematological, neurological and skeletal manifestations in CAAG patients and highlights the importance of an integrated evaluation of these patients. Nevertheless, the nutritional deficiencies in CAAG are largely understudied. We have investigated the frequency and associated features of nutritional deficiencies in CAAG in order to focus on any deficit that may be clinically significant, but relatively easy to correct. This descriptive review updates and summarizes the literature on different nutrient deficiencies in CAAG in order to optimize the treatment and the follow-up of patients affected with CAAG.展开更多
BACKGROUND Calpain-2 is a Ca^2+-dependent cysteine protease,and high calpain-2 activity can enhance apoptosis mediated by multiple triggers.AIM To investigate whether calpain-2 can modulate aberrant endoplasmic reticu...BACKGROUND Calpain-2 is a Ca^2+-dependent cysteine protease,and high calpain-2 activity can enhance apoptosis mediated by multiple triggers.AIM To investigate whether calpain-2 can modulate aberrant endoplasmic reticulum(ER)stress-related apoptosis in rat hepatocyte BRL-3A cells.METHODS BRL-3A cells were treated with varying doses of dithiothreitol(DTT),and their viability and apoptosis were quantified by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-H-tetrazolium bromide and flow cytometry.The expression of ER stress-and apoptosis-related proteins was detected by Western blot analysis.The protease activity of calpain-2 was determined using a fluorescent substrate,Nsuccinyl-Leu-Leu-Val-Tyr-AMC.Intracellular Ca^2+content,and ER and calpain-2 co-localization were characterized by fluorescent microscopy.The impact of calpain-2 silencing by specific small interfering RNA on caspase-12 activation and apoptosis of BRL-3A cells was quantified.RESULTS DTT exhibited dose-dependent cytotoxicity against BRL-3A cells and treatment with 2 mmol/L DTT triggered BRL-3A cell apoptosis.DTT treatment significantly upregulated 78 kDa glucose-regulated protein,activating transcription factor 4,C/EBP-homologous protein expression by>2-fold,and enhanced PRKR-like ER kinase phosphorylation,caspase-12 and caspase-3 cleavage in BRL-3A cells in a trend of time-dependence.DTT treatment also significantly increased intracellular Ca^2+content,calpain-2 expression,and activity by>2-fold in BRL-3A cells.Furthermore,immunofluorescence revealed that DTT treatment promoted the ER accumulation of calpain-2.Moreover,calpain-2 silencing to decrease calpain-2 expression by 85%significantly mitigated DTT-enhanced calpain-2 expression,caspase-12 cleavage,and apoptosis in BRL-3A cells.CONCLUSION The data indicated that Ca^2+-dependent calpain-2 activity promoted the aberrant ER stress-related apoptosis of rat hepatocytes by activating caspase-12 in the ER.展开更多
Calcium is a crucial element for striated muscle function. As such, myoplasmic free Ca2+ concentration is delicately regulated through the concerted action of multiple Ca2+ pathways that relay excitation of the plasma...Calcium is a crucial element for striated muscle function. As such, myoplasmic free Ca2+ concentration is delicately regulated through the concerted action of multiple Ca2+ pathways that relay excitation of the plasma membrane to the intracellular contractile machinery. In skeletal muscle, one of these major Ca2+ pathways is Ca2+ release from intracellular Ca2+ stores through type-1 ryanodine receptor/Ca2+ release channels (RyR1), which positions RyR1 in a strategic cross point to regulate Ca2+ homeostasis. This major Ca2+ traff ic point appears to be highly sensitive to the intracellular environment, which senses through a plethora of chemical and protein-protein interactions. Among these modulators, perhaps one of the most elusive is Triadin, a musclespecif ic protein that is involved in many crucial aspect of muscle function. This family of proteins mediates complex interactions with various Ca2+ modulators and seems poised to be a relevant modulator of Ca2+ signaling in cardiac and skeletal muscles. The purpose of this review is to examine the most recent evidence and current understanding of the role of Triadin in muscle function, in general, with particular emphasis on its contribution to Ca2+ homeostasis.展开更多
基金supported by the National Natural Science Foundation of China,Nos.81671532,81771625(to XF)the Jiangsu Provincial Key Medical Discipline of China,No.ZDXKA2016013(to XF)+3 种基金the Jiangsu Provincial Medical Youth Talent of China,No.QNRC2016758(to XF)the Jiangsu Province Women and Children Health Research Project of China,No.F201750(to XF)the Public Health Technology Project of Suzhou City of China,No.SYS201765(to XF)a grant from the Department of Pediatrics Clinical Center of Suzhou City of China,No.Szzx201504(to XF)。
文摘Transient receptor potential melastatin 2(TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma(PC12) cells injured by oxygen-glucose deprivation(OGD). PC12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reoxygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2(CXCL2), NACHT, LRR, and PYD domain–containing protein 3(NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation.
基金supported by the grants from National Key R&D Program of China (2018YFD0500402)Guangxi Science Foundation for Distinguished Young Scholars (2020GXNSFFA297008)+4 种基金Guangxi Science and Technology Base and Talents Project (AD18281085)Guangxi Natural Science Foundation(2019GXNSFDA245029)Guangxi Hundred-Talent ProgramState Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources(SKLCUSA-a202006)Training Project of High-level Professional and Technical Talents of Guangxi University。
文摘Background:In the livestock industry,intramuscular fat content is a key factor affecting meat quality.Many studies have shown that dietary calcium supplementation is closely related to lipid metabolism.However,few studies have examined the relationship between dietary calcium supplementation and intramuscular fat accumulation.Methods:Here,we used C2C12 cells,C57BL/6 mice(n=8)and three-way cross-breeding pigs(Duroc×Landrace×Large white)(n=10)to study the effect of calcium addition on intramuscular fat accumulation.In vitro,we used calcium chloride to adjust the calcium levels in the medium(2 mmol/L or 3 mmol/L).Then we measured various indicators.In vivo,calcium carbonate was used to regulate calcium levels in feeds(Mice:0.5%calcium or 1.2%calcium)(Pigs:0.9%calcium or 1.5%calcium).Then we tested the mice gastrocnemius muscle triglyceride content,pig longissimus dorsi muscle meat quality and lipidomics.Results:In vitro,calcium addition(3 mmol/L)had no significant effect on cell proliferation,but promoted the differentiation of C2C12 cells into slow-twitch fibers.Calcium supplementation increased triglyceride accumulation in C2C12 cells.Calcium addition increased the number of mitochondria and also increased the calcium level in the mitochondria and reduced the of key enzymes activity involved inβ-oxidation such as acyl-coenzyme A dehydrogenase.Decreasing mitochondrial calcium level can alleviate lipid accumulation induced by calcium addition.In addition,calcium addition also reduced the glycolytic capacity and glycolytic conversion rate of C2C12 cells.In vivo,dietary calcium supplementation(1.2%)promoted the accumulation of triglycerides in the gastrocnemius muscle of mice.Dietary calcium supplementation(1.5%)had no effect on pig weight,but significantly improved the flesh color of the longissimus dorsi muscle,reduced the backfat thickness and increased intramuscular fat content in pigs.Besides,calcium addition had no effect on longissimus dorsi pH,electrical conductivity and shear force.Conclusions:These results suggest that calcium addition promotes intramuscular fat accumulation by inhibiting the oxidation of fatty acids.These findings provide a new tool for increasing intramuscular fat content and an economical strategy for improving meat quality.
文摘The alloy AZ91 containing calcium was prepared under protection of a mixed gas atmosphere of SF6 （1%, volume fraction） and CO2. The added calcium mainly dissolves into the Mg17Al12 phase in the alloy and increases its melting point and the thermal stability. The empirical electron theory （EET） of solid and molecules was used to calculate the valence electron structures （VES） of Mg17Al12 phase with different amounts of calcium additions. The theoretical calculations indicate that calcium dissolved in Mg17Al12 phase increases the strengths of atomic bonds that control the thermal stability of Mg17Al12 phase, and also makes the distribution of valence electrons on the dominant bond network as well as in the whole unit cell of Mg17Al12 more uniform, which are consistent with the experimental results.
基金supported by the National Program on Key Research Project(2016YFA0602900)Nankai University Engineering Research Center for Castor and also supported by"the Fundamental Research Funds for the Central Universities",Nankai University(000082).
文摘A series of diatomite supported Cu/Ni bimetallic catalysts were prepared using the co-impregnation method to improve the efficiency and selectivity toward methyl 12-hydroxystearate in the hydrogenation of methyl ricinoleate.The catalysts were characterized using X-ray diffraction(XRD),transmission electron microscopy(TEM),scanning electron microscopy and energy dispersive X-ray spectroscopy(SEM-EDS),X-ray photoelectron spectroscopy(XPS)and temperature programmed reduction(H2-TPR).All the characterization results verified the formation of highly dispersed Cu/Ni alloy on support.Moreover,by subtly regulating the Ni/Cu molar ratio as well as the reaction parameters,the hydrogenation of methyl ricinoleate to methyl 12-hydroxystearate proceeded efficiently and selectively,affording 97%yield of methyl 12-hydroxystearate and nearly equivalent conversion of methyl ricinoleate under 2 MPa H2 pressure and at 130 C in 4 h with only 1 wt%of the catalyst Ni7Cu1/diatomite(based on methyl ricinoleate).Besides,the supported Cu–Ni bimetallic catalyst is stable during recycle and reuse.After five cycles of reuse,much catalytic activity is still preserved.Therefore,this low-cost and stable bimetallic catalyst would be promising for the hydrogenation of methyl ricinoleate to methyl 12-hydroxystearate,representing an example of green catalysis for efficiently conversion of biomass to value-added chemicals and materials.
文摘Chronic atrophic autoimmune gastritis (CAAG) is an organ-specific autoimmune disease characterized by an immune response, which is directed towards the parietal cells and intrinsic factor of the gastric body and fundus and leads to hypochlorhydria, hypergastrinemia and inadequate production of the intrinsic factor. As a result, the stomach’s secretion of essential substances, such as hydrochloric acid and intrinsic factor, is reduced, leading to digestive impairments. The most common is vitamin B12 deficiency, which results in a megaloblastic anemia and iron malabsorption, leading to iron deficiency anemia. However, in the last years the deficiency of several other vitamins and micronutrients, such as vitamin C, vitamin D, folic acid and calcium, has been increasingly described in patients with CAAG. In addition the occurrence of multiple vitamin deficiencies may lead to severe hematological, neurological and skeletal manifestations in CAAG patients and highlights the importance of an integrated evaluation of these patients. Nevertheless, the nutritional deficiencies in CAAG are largely understudied. We have investigated the frequency and associated features of nutritional deficiencies in CAAG in order to focus on any deficit that may be clinically significant, but relatively easy to correct. This descriptive review updates and summarizes the literature on different nutrient deficiencies in CAAG in order to optimize the treatment and the follow-up of patients affected with CAAG.
基金the National Natural Science Foundation of China,No.81560105the Department of Science and Technology of Guizhou Province,No.LH(2014)7074。
文摘BACKGROUND Calpain-2 is a Ca^2+-dependent cysteine protease,and high calpain-2 activity can enhance apoptosis mediated by multiple triggers.AIM To investigate whether calpain-2 can modulate aberrant endoplasmic reticulum(ER)stress-related apoptosis in rat hepatocyte BRL-3A cells.METHODS BRL-3A cells were treated with varying doses of dithiothreitol(DTT),and their viability and apoptosis were quantified by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-H-tetrazolium bromide and flow cytometry.The expression of ER stress-and apoptosis-related proteins was detected by Western blot analysis.The protease activity of calpain-2 was determined using a fluorescent substrate,Nsuccinyl-Leu-Leu-Val-Tyr-AMC.Intracellular Ca^2+content,and ER and calpain-2 co-localization were characterized by fluorescent microscopy.The impact of calpain-2 silencing by specific small interfering RNA on caspase-12 activation and apoptosis of BRL-3A cells was quantified.RESULTS DTT exhibited dose-dependent cytotoxicity against BRL-3A cells and treatment with 2 mmol/L DTT triggered BRL-3A cell apoptosis.DTT treatment significantly upregulated 78 kDa glucose-regulated protein,activating transcription factor 4,C/EBP-homologous protein expression by>2-fold,and enhanced PRKR-like ER kinase phosphorylation,caspase-12 and caspase-3 cleavage in BRL-3A cells in a trend of time-dependence.DTT treatment also significantly increased intracellular Ca^2+content,calpain-2 expression,and activity by>2-fold in BRL-3A cells.Furthermore,immunofluorescence revealed that DTT treatment promoted the ER accumulation of calpain-2.Moreover,calpain-2 silencing to decrease calpain-2 expression by 85%significantly mitigated DTT-enhanced calpain-2 expression,caspase-12 cleavage,and apoptosis in BRL-3A cells.CONCLUSION The data indicated that Ca^2+-dependent calpain-2 activity promoted the aberrant ER stress-related apoptosis of rat hepatocytes by activating caspase-12 in the ER.
文摘Calcium is a crucial element for striated muscle function. As such, myoplasmic free Ca2+ concentration is delicately regulated through the concerted action of multiple Ca2+ pathways that relay excitation of the plasma membrane to the intracellular contractile machinery. In skeletal muscle, one of these major Ca2+ pathways is Ca2+ release from intracellular Ca2+ stores through type-1 ryanodine receptor/Ca2+ release channels (RyR1), which positions RyR1 in a strategic cross point to regulate Ca2+ homeostasis. This major Ca2+ traff ic point appears to be highly sensitive to the intracellular environment, which senses through a plethora of chemical and protein-protein interactions. Among these modulators, perhaps one of the most elusive is Triadin, a musclespecif ic protein that is involved in many crucial aspect of muscle function. This family of proteins mediates complex interactions with various Ca2+ modulators and seems poised to be a relevant modulator of Ca2+ signaling in cardiac and skeletal muscles. The purpose of this review is to examine the most recent evidence and current understanding of the role of Triadin in muscle function, in general, with particular emphasis on its contribution to Ca2+ homeostasis.
