AIM: To determine the method of growing small intestinal epithelial cells in short-term primary culture and to investigate the effect of extracellular iron concentration ([Fe3+]) on calcium absorption and the relation...AIM: To determine the method of growing small intestinal epithelial cells in short-term primary culture and to investigate the effect of extracellular iron concentration ([Fe3+]) on calcium absorption and the relationship between the rising intracellular calcium concentration ([Ca2+]i) and cell apoptosis in human intestinal epithelial Caco-2 cells. METHODS: Primary culture was used for growing small intestinal epithelial cells. [Ca2+]i was detected by a confocal laser scanning microscope. The changes in [Ca2+]i were represented by fluorescence intensity (FI). The apoptosis was evaluated by flow cytometry. RESULTS: Isolation of epithelial cells and preservation of its three-dimensional integrity were achieved using the digestion technique of a mixture of collagenase Ⅺ and dispase Ⅰ. Purification of the epithelial cells was facilitated by using a simple differential sedimentation method. The results showed that proliferation of normal gut epithelium in vitro was initially dependent upon the maintenance of structural integrity of the tissue. If 0.25% trypsin was used for digestion, the cells were severely damaged and very difficult to stick to the Petri dish for growing. The Fe3+ chelating agent desferrioxamine (100, 200 and 300 μmol/L) increased the FI of Caco-2 cells from 27.50±13.18 (control, n = 150) to 35.71±13.99 (n = 150, P<0.01), 72.19±35.40 (n = 150, P<0.01) and 211.34±29.03 (n = 150,P<0.01) in a concentration-dependent manner. There was a significant decrease in the FI of Caco-2 cells treated by ferric ammonium citrate (FAC, a Fe3+ donor; 10, 50 and 100 μmol/L). The FI value of Caco-2 cells treated by FAC was 185.85±33.77 (n = 150, P<0.01), 122.73±58.47 (n = 150, P<0.01), and 53.29±19.82 (n= 150,P<0.01), respectively, suggesting that calcium absorption was influenced by [Fe3+]. Calcium ionophore A23187(0.1,1.0 and 10 μmol/L) increased the FI of Caco-2 cells from 40.45±13.95 (control, n = 150) to 45.19±21.95 (n = 150, P<0.01), 89.87±43.29 (n = 150, P<0.01) and 104.64±51.07 (n = 150,P<0.01) in a concentration-dependent manner. The positive apoptotic cell number of the Caco-2 cells after being treated with A23187 increased from 0.32% to 0.69%, 0.90% and 1.10%, indicating that the increase in the positive apoptotic cell number was positively correlated with [Ca2+]i. CONCLUSION: Ca2+ absorbability is increased with the decrease of extracellular iron concentration Fe3+ and hindered with the increase of Fe3+ consistence out of them. Furthermore, increase of [Ca2+]i can induce apoptosis in Caco-2 cells.展开更多
Phosvitin(PV)was treated with high-temperature,mild pressure(HTMP),and enzyme combination,and then phosvitin phosphopeptides-calcium(PPP-Ca)complexes were prepared.The low-calcium specific pathogen free-Kunming(SPF-KM...Phosvitin(PV)was treated with high-temperature,mild pressure(HTMP),and enzyme combination,and then phosvitin phosphopeptides-calcium(PPP-Ca)complexes were prepared.The low-calcium specific pathogen free-Kunming(SPF-KM)mice were used to determine the effect of PPP-Ca complexes on intestinal calcium absorption and their utilization for bone formation.The serum calcium content was the highest with the HTMP-Enz-PPP-Ca treatment(2.19 mmol/L),and it significantly down-regulated the abnormal elevation of serum alkaline phosphatase(AKP)caused by calcium deficiency.The low-calcium control group had the lowest calcium deposited to the femur(80.41 mg/g)and the lowest femur bone mineral density(BMD)(0.17 g/cm^(3)),while HTMP-Enz-PPP-Ca significantly improved bone calcium content(94.33 mg/g)and BMD(0.29 g/cm^(3)).The micro-computed tomography(MCT)images showed that the femur with the normal control,PV-Ca,and HTMP-Enz-PPP-Ca treatments had a more compact,complete,and thicker trabecular network than the low-calcium and CaCl_(2)treatments.These results indicated that the organic calcium(HTMP-Enz-PPP-Ca)promoted calcium absorption and bone deposition,and the effect of HTMP-Enz-PPP-Ca was better than the inorganic CaCl_(2).展开更多
Background:Very low-protein(VLP)diets negatively impact calcium(Ca)metabolism and absorption.The objective of this study was to investigate the effect of supplemental branched-chain amino acids(BCAA)and limiting amino...Background:Very low-protein(VLP)diets negatively impact calcium(Ca)metabolism and absorption.The objective of this study was to investigate the effect of supplemental branched-chain amino acids(BCAA)and limiting amino acids(LAA)on Ca digestibility,absorption and reabsorption in pigs fed with VLP diets.Forty-eight piglets were assigned to six treatments:positive control(PC),negative control(NC),and NC containing LAA 25%,LAA 50%,LAA+BCAA 25%(LB25)and LAA+BCAA 50%(LB50)more than recommendations.Results:Relative to PC or NC,LB25 and LB50 had higher digestibility of Ca and plasma Ca and phosphorus(P),but lower plasma vitamin D3.LB50 tended to increase vitamin D receptor transcript and protein in the gut,but decreased mRNA or protein abundance of parathyroid hormone 1 receptor(PTH1R),calbindin 1(CALB1),cytochrome P450 family 27 subfamily B member 1 and occludin in small intestine.LB50 increased the transcript of cytochrome P450 family 24 subfamily A member 1 and PTH1R but decreased the transcript of transient receptor potential cation channel subfamily V member 5,CALB1 and solute carrier family 17 member 4 in kidney.Conclusion:Overall,BCAA increased Ca digestibility through regulating the transcellular and paracellular Ca absorption in the gut and reabsorption in kidney during protein restriction.展开更多
Objective: To study the relationship between Ca 2+ intake,Ca 2+ absorption and Ca 2+ retention. Methods; Thirty-six weaning rats were firstly fed with Ca-poor diet for a 3-week Ca 2+ depletion period,and ...Objective: To study the relationship between Ca 2+ intake,Ca 2+ absorption and Ca 2+ retention. Methods; Thirty-six weaning rats were firstly fed with Ca-poor diet for a 3-week Ca 2+ depletion period,and then randomly divided into 3 groups to receive diet with low,adequate and high Ca 2+ contents.The experiment lasted 12 weeks,with a 3-day metabolism experiment conducted in the 5th week. Results:Supplement with adequate amount of Ca 2+ increased bone Ca 2+,bone mineral density(BMD),bone mineral content(BMC)and the weight of femur(P<0.05).Higher doses of Ca 2+ failed to further improve the efficiency.Analysis showed that Ca 2+ intake was positively correlated to bone Ca 2+,BND and BMC,and negatively correlated to the rates of Ca 2+ absorption and retention.Dose-effect study revealed that the Ca 2+ absorption rate()was a hyperbola function of Ca 2+ intake(X),i.e.=e 6.8068×X -0.8821+20(F=47.3154,P<0.0001).It was found from the results that bone Ca 2+ content(determined by atom absorption spectrophotometer)had a linear correlation to BMC(determined with uni-photon bone mineral densitometer). Conclusion:The rate of Ca 2+ absorption is a hyperbola function of Ca 2+ intake.The increment of Ca 2+ absorption will be minimal when Ca 2+ intake exceeds 3d 250 mg in rats.BMC,better than BMD,may substitute for the ideal bone Ca 2+ content as the index to assess the efficacy of Ca 2+ supplement.展开更多
Dear Editor,Postmenopausal estrogen deficiency in humans is thought to be the main cause of osteoporosis and diminished calcium absorption in older women.Estrogen supplementation may alleviate the problem of osteoporo...Dear Editor,Postmenopausal estrogen deficiency in humans is thought to be the main cause of osteoporosis and diminished calcium absorption in older women.Estrogen supplementation may alleviate the problem of osteoporosis in estrogen-deficient women by promoting calcium absorption.展开更多
In a rotary kiln process for phosphoric acid production,the reaction between gaseous phosphorus pentoxide(P2O5)and phosphate ore and silica contained in feed balls(the so-called P2O5"absorption")not only reduces...In a rotary kiln process for phosphoric acid production,the reaction between gaseous phosphorus pentoxide(P2O5)and phosphate ore and silica contained in feed balls(the so-called P2O5"absorption")not only reduces phosphorous recovery but also generates a large amount of low melting-point side products.The products may give rise to formation of kiln ring,which interferes with kiln operation performance.In this study,the reactions of gaseous P2O5with solid calcium phosphate(Ca3(PO4)2),silica(SiO2)and their mixture,respectively,were investigated via combined chemical analysis and various characterizations comprised of X-ray diffraction(XRD),Fourier-transform infrared(FT-IR)spectroscopy,thermogravimetric analysis and differential scanning calorimeter(TG&DSC),and scanning electron microscopy and energy dispersive spectrometer(SEM&EDS).Attentions were focused on apparent morphology,phase transformation and thermal stability of the products of the P2O5"absorption"at different temperatures.The results show that the temperature significantly affected the"absorption".The reaction between pure Ca3(PO4)2 and P2O5 occurred at 500℃.Calcium metaphosphate(Ca(PO3)2)was the primary product at the temperatures≤900℃ with its melting point≤900℃ while calcium pyrophosphate(Ca2P2O7)was obtained over 1000℃,which has a melting point≤1200℃.The"absorption"by pure SiO2 started at 800℃ and the most significant reaction occurred at 1000℃ with formation of silicon pyrophosphate(SiP2O7)product of melting point≤1000℃.Using mixed Ca3(PO4)2and SiO2as raw material,the"absorption"by Ca3(PO4)2 was enhanced due to existence of silica.At 600–700℃,silica was inert to P2O5and thus formed a porous structure in the raw material,which accelerated diffusion of gaseous P2O5inside the mixture.At higher temperatures,the combined"absorption"by calcium phosphate and reaction between silicon dioxide and the"absorption"product calcium pyrophosphate,reinforced the"absorption"by the mixture.Besides,it was found that both Ca(PO3)2and SiP2O7were unstable at high temperatures and would decompose to Ca2P2O7and SiO2,respectively,at over 1000℃ and 1100℃ with the release of gaseous P2O5at the same time.展开更多
The objective of this work was to evaluate the influence of bread with thermal treated lentil flour on metabolic and physiological parameters.Thirty-two Wistar rats fed for 60 days with different diets were studied:co...The objective of this work was to evaluate the influence of bread with thermal treated lentil flour on metabolic and physiological parameters.Thirty-two Wistar rats fed for 60 days with different diets were studied:control(C),wheat bread(WB),wheat-lentil bread(LB),and lentil flour(LF).Food intake,weight gain,feces moisture,and apparent calcium absorption(AACa)were recorded during the experiment.The weight and pH of the cecum,as well as serum parameters,were evaluated after sacrifice.LB group showed higher intake and weight gain.Diets with lentil produced several positive effects:an increase in feces moisture,higher AACa,augmentation of cecum weight,and acidification in the cecum.Besides,an improvement in bone quality and a reduction of cholesterol was observed with lentil diets.Because of the named benefits,bread containing lentil flour could be considered a functional food.展开更多
基金Supported by the Natural Science Foundation of Hebei Province,No. 303158 Education Department Foundation of Hebei Province,No. 2002136
文摘AIM: To determine the method of growing small intestinal epithelial cells in short-term primary culture and to investigate the effect of extracellular iron concentration ([Fe3+]) on calcium absorption and the relationship between the rising intracellular calcium concentration ([Ca2+]i) and cell apoptosis in human intestinal epithelial Caco-2 cells. METHODS: Primary culture was used for growing small intestinal epithelial cells. [Ca2+]i was detected by a confocal laser scanning microscope. The changes in [Ca2+]i were represented by fluorescence intensity (FI). The apoptosis was evaluated by flow cytometry. RESULTS: Isolation of epithelial cells and preservation of its three-dimensional integrity were achieved using the digestion technique of a mixture of collagenase Ⅺ and dispase Ⅰ. Purification of the epithelial cells was facilitated by using a simple differential sedimentation method. The results showed that proliferation of normal gut epithelium in vitro was initially dependent upon the maintenance of structural integrity of the tissue. If 0.25% trypsin was used for digestion, the cells were severely damaged and very difficult to stick to the Petri dish for growing. The Fe3+ chelating agent desferrioxamine (100, 200 and 300 μmol/L) increased the FI of Caco-2 cells from 27.50±13.18 (control, n = 150) to 35.71±13.99 (n = 150, P<0.01), 72.19±35.40 (n = 150, P<0.01) and 211.34±29.03 (n = 150,P<0.01) in a concentration-dependent manner. There was a significant decrease in the FI of Caco-2 cells treated by ferric ammonium citrate (FAC, a Fe3+ donor; 10, 50 and 100 μmol/L). The FI value of Caco-2 cells treated by FAC was 185.85±33.77 (n = 150, P<0.01), 122.73±58.47 (n = 150, P<0.01), and 53.29±19.82 (n= 150,P<0.01), respectively, suggesting that calcium absorption was influenced by [Fe3+]. Calcium ionophore A23187(0.1,1.0 and 10 μmol/L) increased the FI of Caco-2 cells from 40.45±13.95 (control, n = 150) to 45.19±21.95 (n = 150, P<0.01), 89.87±43.29 (n = 150, P<0.01) and 104.64±51.07 (n = 150,P<0.01) in a concentration-dependent manner. The positive apoptotic cell number of the Caco-2 cells after being treated with A23187 increased from 0.32% to 0.69%, 0.90% and 1.10%, indicating that the increase in the positive apoptotic cell number was positively correlated with [Ca2+]i. CONCLUSION: Ca2+ absorbability is increased with the decrease of extracellular iron concentration Fe3+ and hindered with the increase of Fe3+ consistence out of them. Furthermore, increase of [Ca2+]i can induce apoptosis in Caco-2 cells.
基金supported by the General Program of National Natural Science Foundation of China(32072237)the Hubei Provincial Natural Science Foundation of China(2020CFB583)Fundamental Research Funds for the Central Universities(2662020SPPY006)。
文摘Phosvitin(PV)was treated with high-temperature,mild pressure(HTMP),and enzyme combination,and then phosvitin phosphopeptides-calcium(PPP-Ca)complexes were prepared.The low-calcium specific pathogen free-Kunming(SPF-KM)mice were used to determine the effect of PPP-Ca complexes on intestinal calcium absorption and their utilization for bone formation.The serum calcium content was the highest with the HTMP-Enz-PPP-Ca treatment(2.19 mmol/L),and it significantly down-regulated the abnormal elevation of serum alkaline phosphatase(AKP)caused by calcium deficiency.The low-calcium control group had the lowest calcium deposited to the femur(80.41 mg/g)and the lowest femur bone mineral density(BMD)(0.17 g/cm^(3)),while HTMP-Enz-PPP-Ca significantly improved bone calcium content(94.33 mg/g)and BMD(0.29 g/cm^(3)).The micro-computed tomography(MCT)images showed that the femur with the normal control,PV-Ca,and HTMP-Enz-PPP-Ca treatments had a more compact,complete,and thicker trabecular network than the low-calcium and CaCl_(2)treatments.These results indicated that the organic calcium(HTMP-Enz-PPP-Ca)promoted calcium absorption and bone deposition,and the effect of HTMP-Enz-PPP-Ca was better than the inorganic CaCl_(2).
基金funded by Animal Health and Production and Animal Products:Improved Nutritional Performance,Growth,and Lactation of Animals,grant no. 2021–67016-33399/project accession no. 1024334Hatch project accession no. 1012889 from the USDA National Institute of Food and Agriculture (NIFA)
文摘Background:Very low-protein(VLP)diets negatively impact calcium(Ca)metabolism and absorption.The objective of this study was to investigate the effect of supplemental branched-chain amino acids(BCAA)and limiting amino acids(LAA)on Ca digestibility,absorption and reabsorption in pigs fed with VLP diets.Forty-eight piglets were assigned to six treatments:positive control(PC),negative control(NC),and NC containing LAA 25%,LAA 50%,LAA+BCAA 25%(LB25)and LAA+BCAA 50%(LB50)more than recommendations.Results:Relative to PC or NC,LB25 and LB50 had higher digestibility of Ca and plasma Ca and phosphorus(P),but lower plasma vitamin D3.LB50 tended to increase vitamin D receptor transcript and protein in the gut,but decreased mRNA or protein abundance of parathyroid hormone 1 receptor(PTH1R),calbindin 1(CALB1),cytochrome P450 family 27 subfamily B member 1 and occludin in small intestine.LB50 increased the transcript of cytochrome P450 family 24 subfamily A member 1 and PTH1R but decreased the transcript of transient receptor potential cation channel subfamily V member 5,CALB1 and solute carrier family 17 member 4 in kidney.Conclusion:Overall,BCAA increased Ca digestibility through regulating the transcellular and paracellular Ca absorption in the gut and reabsorption in kidney during protein restriction.
文摘Objective: To study the relationship between Ca 2+ intake,Ca 2+ absorption and Ca 2+ retention. Methods; Thirty-six weaning rats were firstly fed with Ca-poor diet for a 3-week Ca 2+ depletion period,and then randomly divided into 3 groups to receive diet with low,adequate and high Ca 2+ contents.The experiment lasted 12 weeks,with a 3-day metabolism experiment conducted in the 5th week. Results:Supplement with adequate amount of Ca 2+ increased bone Ca 2+,bone mineral density(BMD),bone mineral content(BMC)and the weight of femur(P<0.05).Higher doses of Ca 2+ failed to further improve the efficiency.Analysis showed that Ca 2+ intake was positively correlated to bone Ca 2+,BND and BMC,and negatively correlated to the rates of Ca 2+ absorption and retention.Dose-effect study revealed that the Ca 2+ absorption rate()was a hyperbola function of Ca 2+ intake(X),i.e.=e 6.8068×X -0.8821+20(F=47.3154,P<0.0001).It was found from the results that bone Ca 2+ content(determined by atom absorption spectrophotometer)had a linear correlation to BMC(determined with uni-photon bone mineral densitometer). Conclusion:The rate of Ca 2+ absorption is a hyperbola function of Ca 2+ intake.The increment of Ca 2+ absorption will be minimal when Ca 2+ intake exceeds 3d 250 mg in rats.BMC,better than BMD,may substitute for the ideal bone Ca 2+ content as the index to assess the efficacy of Ca 2+ supplement.
基金supported by the National Key Research and Development Program of China (2023YFD1301003)the China Agricultural Research System program (CARS-40K08)。
文摘Dear Editor,Postmenopausal estrogen deficiency in humans is thought to be the main cause of osteoporosis and diminished calcium absorption in older women.Estrogen supplementation may alleviate the problem of osteoporosis in estrogen-deficient women by promoting calcium absorption.
基金Suported by the breeding programs of Sichuan University(SCU2015C002)
文摘In a rotary kiln process for phosphoric acid production,the reaction between gaseous phosphorus pentoxide(P2O5)and phosphate ore and silica contained in feed balls(the so-called P2O5"absorption")not only reduces phosphorous recovery but also generates a large amount of low melting-point side products.The products may give rise to formation of kiln ring,which interferes with kiln operation performance.In this study,the reactions of gaseous P2O5with solid calcium phosphate(Ca3(PO4)2),silica(SiO2)and their mixture,respectively,were investigated via combined chemical analysis and various characterizations comprised of X-ray diffraction(XRD),Fourier-transform infrared(FT-IR)spectroscopy,thermogravimetric analysis and differential scanning calorimeter(TG&DSC),and scanning electron microscopy and energy dispersive spectrometer(SEM&EDS).Attentions were focused on apparent morphology,phase transformation and thermal stability of the products of the P2O5"absorption"at different temperatures.The results show that the temperature significantly affected the"absorption".The reaction between pure Ca3(PO4)2 and P2O5 occurred at 500℃.Calcium metaphosphate(Ca(PO3)2)was the primary product at the temperatures≤900℃ with its melting point≤900℃ while calcium pyrophosphate(Ca2P2O7)was obtained over 1000℃,which has a melting point≤1200℃.The"absorption"by pure SiO2 started at 800℃ and the most significant reaction occurred at 1000℃ with formation of silicon pyrophosphate(SiP2O7)product of melting point≤1000℃.Using mixed Ca3(PO4)2and SiO2as raw material,the"absorption"by Ca3(PO4)2 was enhanced due to existence of silica.At 600–700℃,silica was inert to P2O5and thus formed a porous structure in the raw material,which accelerated diffusion of gaseous P2O5inside the mixture.At higher temperatures,the combined"absorption"by calcium phosphate and reaction between silicon dioxide and the"absorption"product calcium pyrophosphate,reinforced the"absorption"by the mixture.Besides,it was found that both Ca(PO3)2and SiP2O7were unstable at high temperatures and would decompose to Ca2P2O7and SiO2,respectively,at over 1000℃ and 1100℃ with the release of gaseous P2O5at the same time.
文摘The objective of this work was to evaluate the influence of bread with thermal treated lentil flour on metabolic and physiological parameters.Thirty-two Wistar rats fed for 60 days with different diets were studied:control(C),wheat bread(WB),wheat-lentil bread(LB),and lentil flour(LF).Food intake,weight gain,feces moisture,and apparent calcium absorption(AACa)were recorded during the experiment.The weight and pH of the cecum,as well as serum parameters,were evaluated after sacrifice.LB group showed higher intake and weight gain.Diets with lentil produced several positive effects:an increase in feces moisture,higher AACa,augmentation of cecum weight,and acidification in the cecum.Besides,an improvement in bone quality and a reduction of cholesterol was observed with lentil diets.Because of the named benefits,bread containing lentil flour could be considered a functional food.