Background Commercial diets are frequently formulated to meet or exceed nutrient levels including those of lim-iting essential amino acids(AA)covering potential individual variations within the herd.However,the provis...Background Commercial diets are frequently formulated to meet or exceed nutrient levels including those of lim-iting essential amino acids(AA)covering potential individual variations within the herd.However,the provision of dietary excess of AA,such as Lys,may lead to reduced appetite and growth in pigs.The mechanisms modulat-ing these responses have not been extensively investigated.This study evaluated the effect of Lys dietary excesses on performance and satiety biomarkers in post weaning pigs.Methods Twenty-four pigs aged 21 d and weighing 6.81±0.12kg(mean±SEM)were individually housed and offered 1 of 4 dietary treatments for 3weeks:a diet containing a standardized ileal digestible Lys reaching 100%(T0),120%(T1),150%(T2)or 200%(T3)of the NRC(2012)requirements.At the end of the experiment,blood samples from the cephalic vein of the T0 and T3 groups were obtained for AA analysis.In addition,primary intestinal cultures from T0 pigs were used,following their humane killing,to evaluate the effect of Lys on gut hormone secretion and AA sensors gene expression under ex vivo conditions.Results Feed intake was linearly reduced(P<0.001)and the weight gain to feed ratio reduced(P<0.10)with increased dietary levels of Lys during the third-and first-week post weaning,respectively.Cholecystokinin con-centration(P<0.05)and the metabotropic glutamate receptor 1 and the solute carrier family 7 member 2(P<0.10)gene expression was enhanced in proximal jejunum tissues incubated with Lys at 20mmol/L when compared to the control(Lys 0mmol/L).Plasma Lys and Glu(P<0.05)concentration increased in the T3 compared to T0 pigs.In contrast,plasma levels of His,Val,Thr,Leu(P<0.05)and Gln(P<0.10)were lower in T3 than T0 pigs.Conclusion The present results confirm that excess dietary Lys inhibits hunger in pigs.Moreover,the results provide evidence of pre-and post-absorptive mechanisms modulating these responses.Lys dietary excesses should be nar-rowed,when possible,to avoid negative effects of the AA on appetite in pigs.展开更多
[Objectives] In this study,routine blood indexes of Guike pig were detected to clarify quantity variation and morphology distribution of different functional cells（ red blood cells,white blood cells and platelets）,t...[Objectives] In this study,routine blood indexes of Guike pig were detected to clarify quantity variation and morphology distribution of different functional cells（ red blood cells,white blood cells and platelets）,thus providing reference data for immunological evaluation and disease resistance research of Guike pig. [Methods] Blood samples were collected from forty-four 49-day-old healthy Guike pigs（ 22 males and 22 females） once a week for seven consecutive weeks to detect routine blood indexes,and the results were compared with routine blood indexes of Landrace pig and Enshi black pig. [Results]Among red blood cell indexes of Guike pig at different day ages,red blood cell count（ RBC） and hemoglobin count（ HGB） both reached the lowest levels（ 6. 85 × 10^12/L and 97. 1 g/L）at 70 days of age,which were significantly lower than the corresponding indexes at 49,56,63,84 and 91 days of age（ P 〈 0. 05,similarly hereinafter）; mean corpuscular hemoglobin（ MCH） and mean corpuscular hemoglobin concentration（ MCHC） demonstrated basically the same variation pattern,which both reached the lowest levels（ 15. 11 pg and 275. 70 g/L） at 70 days of age and reached the highest levels（ 16. 40 pg and 314. 30 g/L） at 91 days of age; hematocrit（ HCT）,mean corpuscular volume（ MCV） and red blood cell distribution width（ RDW） exhibited relatively stable changes with no significant differences among different day ages（ P 〉 0. 05,similarly hereinafter）. Among white blood cell indexes of Guike pig at different day ages,white blood cell count（ WBC） reached the lowest level（ 13. 71 × 10^9/L） at 63 days of age and reached the highest level（ 17. 52 × 10^9/L） at 84 days of age,with significant differences; neutrophil count（ NEUT）reached the lowest level（ 3. 61 × 10^9/L） at 70 days of age,which was significantly lower than that at 49,56 and 77 days of age; lymphocyte count（ LYMPH） exhibited relatively stable changes and was not significantly affected by the increase in day age. Among platelet indexes of Guike pig at different day ages,platelet count（ PLT）,thrombocytocrit（ PCT） and platelet volume distribution width（ PDW） exhibited relatively stable changes among different day ages,which were not significantly affected by the increase in day age. Compared to 35-day-old Landrace pig and 5-month-old Enshi black pig,most routine blood indexes of Guike pig were similar to or ranged between those of Landrace pig and Enshi black pig except HCT,mean platelet volume（ MPV）,PCT and PDW. [Conclusions]Guike pig has a certain uniqueness in routine blood indexes,which has developed a stable immune system at 49-91 days of age.展开更多
The acute （15 minutes-4 hours） effects of exposure to the inhaled DEE （diesel exhaust emitted） on blood parameters of Guinea pigs Cavia porcellus were assessed via CBC （complete blood count） as an indicator usin...The acute （15 minutes-4 hours） effects of exposure to the inhaled DEE （diesel exhaust emitted） on blood parameters of Guinea pigs Cavia porcellus were assessed via CBC （complete blood count） as an indicator using an automated blood analyzer machines （Cell DYN 1700）. The results showed significant changes in some parameters in comparison with control animals started as early as 15 minutes and intensified by the continuous exposure time, leading to significant increase （p 〈 0.05） in the number of both WBC （white blood cells）, PLT （blood platelets） and decrease in numbers of RBC （red blood corpuscles） while hemoglobin percentages （Hb%） showed an insignificant （p 〉 0.05） fluctuating rhythm. The percentages of both Neutrophil and Lymphocyt recorded a significant increase （p 〈 0.05） 1 hour post-exposure to the DEE while other WBCs showed less changes. It is concluded that blood components are so sensitive to DEE and do respond quicker to inhaled pollutants via changes in some components of WBC% but decrease in RBC% and the likelihood of blood clotting due to increase in blood platelets. These results indicate the acute toxic risks of the exposure to DEE of mechanics and technicians who work in vicinity of DEE sources or/and spend most daily hours in semi closed areas.展开更多
[Objectives]This study aimed to optimize the chelation process for complex microelement iron supplement derived from pig blood by response surface methodology.[Methods]On the basis of single-factor test,p H value,conc...[Objectives]This study aimed to optimize the chelation process for complex microelement iron supplement derived from pig blood by response surface methodology.[Methods]On the basis of single-factor test,p H value,concentration of polypeptide solution and volume ratio of polypeptide solution to FeCl_2 solution were selected as influencing factors with Fe(II)chelation rate as the indicator for Box-Behnken central composite experimental design with three factors and three levels.The effects of three factors on the response value were analyzed by response surface methodology.[Results]The optimized chelation process for complex microelement iron supplement derived from pig blood by response surface methodology was as follows:pH 5.40,polypeptide solution concentration 2.27%,volume ratio of polypeptide solution to FeCl_2 solution 2.16∶1.Under this condition,the predictive Fe(II)chelation rate of iron supplement was 79.37%,while the actual value was 79.41%.[Conclusions]The optimized process may provide new thoughts for the development and utilization of complex microelement iron supplement derived from pig blood.展开更多
Objective: TO investigate the killing differences of human blood group antibodies IgG and IgM on PBMC and RBC in peripheral blood of genetically modified pigs, and provide theoretical basis for the selection of pig bl...Objective: TO investigate the killing differences of human blood group antibodies IgG and IgM on PBMC and RBC in peripheral blood of genetically modified pigs, and provide theoretical basis for the selection of pig blood group for clinical application of porcine xenotransplantation. Methods: Serum samples were collected from 20 healthy subjects, 20 patients with end‑stage renal disease and 20 brain dead organ donors, and divided into 4 groups on the basis of ABO blood group (A: n=20;B: n=17;AB: n=7;O: n=16). Flow cytometry was used to detect antibody binding or complement dependent cytotoxicity test (CDC) between human serum and O blood group Wild‑type (WT), α1, 3‑galactosyltransferase gene‑knockout(GTKO), cytidine monophosphate‑N‑acetylneuraminic acid hydroxylase gene‑knockout (GTKO/β4GalNT2KO), β‑1, 4N‑acetylgalactosaminyltransferase gene‑knockout(GTKO/CMAHKO) and mononuclear cells (PBMC) and red blood cells (RBC) of Triple knockout (TKO/hCD55) porcine peripheral blood, respectively. Results: There was no significant difference in binding and killing of human serum antibodies with blood group A, B, AB and O on PBMC of WT pigs, GTKO pigs, GTKO/β4GalNT2KO pigs, GTKO/CMAHKO pigs and TKO/hCD55 pigs, respectively. There was no significant difference in RBC binding with RBC of WT pig, GTKO pig, GTKO/β4GalNT2KO pig, GTKO/CMAHKO pig and TKO/hCD55 pig. Conclusion: The selection of recipients of pigs with type O blood group can be done without considering blood group.展开更多
Objective To evaluate the effect of wound closure tension on the blood flow of the expanded pedicled fasciocutaneous flap,so as to find the best tension for the blood supply of the flap.Methods 8 piglets,aged 9-12 mon...Objective To evaluate the effect of wound closure tension on the blood flow of the expanded pedicled fasciocutaneous flap,so as to find the best tension for the blood supply of the flap.Methods 8 piglets,aged 9-12 months,were used.On展开更多
There is a critical shortage of organs, cells, and corneas from deceased human donors worldwide. There are also shortages of human blood for transfusion. A potential solution to all of these problems is the transplant...There is a critical shortage of organs, cells, and corneas from deceased human donors worldwide. There are also shortages of human blood for transfusion. A potential solution to all of these problems is the transplantation of organs, cells, and corneas from a readily available animal species, such as the pig, and the transfusion of red blood cells from pigs into humans. However, to achieve these ends, major immunologic and other barriers have to be overcome. Considerable progress has been made in this respect by the genetic modification of pigs to protect their tissues from the primate immune response and to correct several molecular incompatibilities that exist between pig and primate. These have included knockout of genes responsible for the expression of major antigenic targets for primate natural anti-pig antibodies, insertion of human complement- and coagulation-regulatory transgenes, and knockdown of swine leukocyte antigens that stimulate the primate＇s adaptive immune response. As a result of these manipulations, the administration of novel immunosuppressive agents, and other innovations, pig hearts have now functioned in baboons for 6-8 months, pig islets have maintained normoglycemia in diabetic monkeys for 〉 1 year, and pig corneas have maintained transparency for several months. Clinical trials of pig islet trans- plantation are already in progress. Future developments will involve further genetic manipulations of the organ- source pig, with most of the genes that are likely to be beneficial already identified.展开更多
We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy...We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy for neurodegenerative diseases,neurotrauma,and stroke.In this study,using a mini pig model of spinal cord injury,we proposed for the first time the use of gene-modified leucoconcentrate prepared from peripheral blood in the plastic blood bag for personalized ex vivo gene therapy.Leucoconcentrate obtained from mini pig peripheral blood was transduced with a chimeric adenoviral vector(Ad5/35 F)that carried an enhanced green fluorescent protein(EGFP)reporter gene in the plastic blood bag.The day after blood donation,the mini pigs were subjected to moderate SCI and four hours post-surgery they were intravenously autoinfused with gene-modified leucoconcentrate.A week after gene-modified leucoconcentrate therapy,fluorescent microscopy revealed EGFP-expressing leucocytes in spinal cord at the site of contusion injury.In the spleen the groups of EGFP-positive cells located in the lymphoid follicles were observed.In vitro flow cytometry and fluorescent microscopy studies of the gene-modified leucoconcentrate samples also confirmed the production of EGFP by leucocytes.Thus,the efficacy of leucocytes transduction in the plastic blood bag and their migratory potential suggest their use for temporary production of recombinant biologically active molecules to correct certain pathological conditions.This paper presents a proof-of-concept of simple,safe and effective approach for personalized ex vivo gene therapy based on gene-modified leucoconcentrate autoinfusion.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.5)on May 27,2014.展开更多
基金This study was partially supported by The University of Queensland and Australian Pork Limited as part of the project APL 2016/053.
文摘Background Commercial diets are frequently formulated to meet or exceed nutrient levels including those of lim-iting essential amino acids(AA)covering potential individual variations within the herd.However,the provision of dietary excess of AA,such as Lys,may lead to reduced appetite and growth in pigs.The mechanisms modulat-ing these responses have not been extensively investigated.This study evaluated the effect of Lys dietary excesses on performance and satiety biomarkers in post weaning pigs.Methods Twenty-four pigs aged 21 d and weighing 6.81±0.12kg(mean±SEM)were individually housed and offered 1 of 4 dietary treatments for 3weeks:a diet containing a standardized ileal digestible Lys reaching 100%(T0),120%(T1),150%(T2)or 200%(T3)of the NRC(2012)requirements.At the end of the experiment,blood samples from the cephalic vein of the T0 and T3 groups were obtained for AA analysis.In addition,primary intestinal cultures from T0 pigs were used,following their humane killing,to evaluate the effect of Lys on gut hormone secretion and AA sensors gene expression under ex vivo conditions.Results Feed intake was linearly reduced(P<0.001)and the weight gain to feed ratio reduced(P<0.10)with increased dietary levels of Lys during the third-and first-week post weaning,respectively.Cholecystokinin con-centration(P<0.05)and the metabotropic glutamate receptor 1 and the solute carrier family 7 member 2(P<0.10)gene expression was enhanced in proximal jejunum tissues incubated with Lys at 20mmol/L when compared to the control(Lys 0mmol/L).Plasma Lys and Glu(P<0.05)concentration increased in the T3 compared to T0 pigs.In contrast,plasma levels of His,Val,Thr,Leu(P<0.05)and Gln(P<0.10)were lower in T3 than T0 pigs.Conclusion The present results confirm that excess dietary Lys inhibits hunger in pigs.Moreover,the results provide evidence of pre-and post-absorptive mechanisms modulating these responses.Lys dietary excesses should be nar-rowed,when possible,to avoid negative effects of the AA on appetite in pigs.
基金Supported by Major Scientific and Technological Special Project of Guangxi Zhuang Autonomous Region"Innovation and Demonstration of Ecological Comprehensive Prevention and Control Technology for Major Diseases of Livestock and Poultry"(GKAA17204057)Natural Science Foundation of Guangxi Zhuang Autonomous Region(2015GXNSFAA139070)+1 种基金Open-end Fund for Key Laboratory of Livestock Genetic Improvement of Guangxi Zhuang Autonomous Region(2014GXKLLGI-07)Project of Aquatic Animal Husbandry Science and Technology Extension and Application of Guangxi Zhuang Autonomous Region(GYMK201528030,GYMK201633034)
文摘[Objectives] In this study,routine blood indexes of Guike pig were detected to clarify quantity variation and morphology distribution of different functional cells（ red blood cells,white blood cells and platelets）,thus providing reference data for immunological evaluation and disease resistance research of Guike pig. [Methods] Blood samples were collected from forty-four 49-day-old healthy Guike pigs（ 22 males and 22 females） once a week for seven consecutive weeks to detect routine blood indexes,and the results were compared with routine blood indexes of Landrace pig and Enshi black pig. [Results]Among red blood cell indexes of Guike pig at different day ages,red blood cell count（ RBC） and hemoglobin count（ HGB） both reached the lowest levels（ 6. 85 × 10^12/L and 97. 1 g/L）at 70 days of age,which were significantly lower than the corresponding indexes at 49,56,63,84 and 91 days of age（ P 〈 0. 05,similarly hereinafter）; mean corpuscular hemoglobin（ MCH） and mean corpuscular hemoglobin concentration（ MCHC） demonstrated basically the same variation pattern,which both reached the lowest levels（ 15. 11 pg and 275. 70 g/L） at 70 days of age and reached the highest levels（ 16. 40 pg and 314. 30 g/L） at 91 days of age; hematocrit（ HCT）,mean corpuscular volume（ MCV） and red blood cell distribution width（ RDW） exhibited relatively stable changes with no significant differences among different day ages（ P 〉 0. 05,similarly hereinafter）. Among white blood cell indexes of Guike pig at different day ages,white blood cell count（ WBC） reached the lowest level（ 13. 71 × 10^9/L） at 63 days of age and reached the highest level（ 17. 52 × 10^9/L） at 84 days of age,with significant differences; neutrophil count（ NEUT）reached the lowest level（ 3. 61 × 10^9/L） at 70 days of age,which was significantly lower than that at 49,56 and 77 days of age; lymphocyte count（ LYMPH） exhibited relatively stable changes and was not significantly affected by the increase in day age. Among platelet indexes of Guike pig at different day ages,platelet count（ PLT）,thrombocytocrit（ PCT） and platelet volume distribution width（ PDW） exhibited relatively stable changes among different day ages,which were not significantly affected by the increase in day age. Compared to 35-day-old Landrace pig and 5-month-old Enshi black pig,most routine blood indexes of Guike pig were similar to or ranged between those of Landrace pig and Enshi black pig except HCT,mean platelet volume（ MPV）,PCT and PDW. [Conclusions]Guike pig has a certain uniqueness in routine blood indexes,which has developed a stable immune system at 49-91 days of age.
文摘The acute （15 minutes-4 hours） effects of exposure to the inhaled DEE （diesel exhaust emitted） on blood parameters of Guinea pigs Cavia porcellus were assessed via CBC （complete blood count） as an indicator using an automated blood analyzer machines （Cell DYN 1700）. The results showed significant changes in some parameters in comparison with control animals started as early as 15 minutes and intensified by the continuous exposure time, leading to significant increase （p 〈 0.05） in the number of both WBC （white blood cells）, PLT （blood platelets） and decrease in numbers of RBC （red blood corpuscles） while hemoglobin percentages （Hb%） showed an insignificant （p 〉 0.05） fluctuating rhythm. The percentages of both Neutrophil and Lymphocyt recorded a significant increase （p 〈 0.05） 1 hour post-exposure to the DEE while other WBCs showed less changes. It is concluded that blood components are so sensitive to DEE and do respond quicker to inhaled pollutants via changes in some components of WBC% but decrease in RBC% and the likelihood of blood clotting due to increase in blood platelets. These results indicate the acute toxic risks of the exposure to DEE of mechanics and technicians who work in vicinity of DEE sources or/and spend most daily hours in semi closed areas.
基金Supported by Youth Fund of National Natural Science Foundation of China(31801673)Talent Development Fund of Anhui Academy of Agricultural Sciences(17F1205)+2 种基金Youth Innovation Fund of President of Anhui Academy of Agricultural Sciences(17B1220)Team Building Project of Anhui Academy of Agricultural Sciences(18C1225)Youth Fund of Natural Science Foundation of Anhui Province(1808085QC94)
文摘[Objectives]This study aimed to optimize the chelation process for complex microelement iron supplement derived from pig blood by response surface methodology.[Methods]On the basis of single-factor test,p H value,concentration of polypeptide solution and volume ratio of polypeptide solution to FeCl_2 solution were selected as influencing factors with Fe(II)chelation rate as the indicator for Box-Behnken central composite experimental design with three factors and three levels.The effects of three factors on the response value were analyzed by response surface methodology.[Results]The optimized chelation process for complex microelement iron supplement derived from pig blood by response surface methodology was as follows:pH 5.40,polypeptide solution concentration 2.27%,volume ratio of polypeptide solution to FeCl_2 solution 2.16∶1.Under this condition,the predictive Fe(II)chelation rate of iron supplement was 79.37%,while the actual value was 79.41%.[Conclusions]The optimized process may provide new thoughts for the development and utilization of complex microelement iron supplement derived from pig blood.
基金Key Science and Technology Project of Hainan Provincial Science and Technology Department (ZDKT2019009)Hainan Clinical Medical Center construction project.
文摘Objective: TO investigate the killing differences of human blood group antibodies IgG and IgM on PBMC and RBC in peripheral blood of genetically modified pigs, and provide theoretical basis for the selection of pig blood group for clinical application of porcine xenotransplantation. Methods: Serum samples were collected from 20 healthy subjects, 20 patients with end‑stage renal disease and 20 brain dead organ donors, and divided into 4 groups on the basis of ABO blood group (A: n=20;B: n=17;AB: n=7;O: n=16). Flow cytometry was used to detect antibody binding or complement dependent cytotoxicity test (CDC) between human serum and O blood group Wild‑type (WT), α1, 3‑galactosyltransferase gene‑knockout(GTKO), cytidine monophosphate‑N‑acetylneuraminic acid hydroxylase gene‑knockout (GTKO/β4GalNT2KO), β‑1, 4N‑acetylgalactosaminyltransferase gene‑knockout(GTKO/CMAHKO) and mononuclear cells (PBMC) and red blood cells (RBC) of Triple knockout (TKO/hCD55) porcine peripheral blood, respectively. Results: There was no significant difference in binding and killing of human serum antibodies with blood group A, B, AB and O on PBMC of WT pigs, GTKO pigs, GTKO/β4GalNT2KO pigs, GTKO/CMAHKO pigs and TKO/hCD55 pigs, respectively. There was no significant difference in RBC binding with RBC of WT pig, GTKO pig, GTKO/β4GalNT2KO pig, GTKO/CMAHKO pig and TKO/hCD55 pig. Conclusion: The selection of recipients of pigs with type O blood group can be done without considering blood group.
文摘Objective To evaluate the effect of wound closure tension on the blood flow of the expanded pedicled fasciocutaneous flap,so as to find the best tension for the blood supply of the flap.Methods 8 piglets,aged 9-12 months,were used.On
文摘There is a critical shortage of organs, cells, and corneas from deceased human donors worldwide. There are also shortages of human blood for transfusion. A potential solution to all of these problems is the transplantation of organs, cells, and corneas from a readily available animal species, such as the pig, and the transfusion of red blood cells from pigs into humans. However, to achieve these ends, major immunologic and other barriers have to be overcome. Considerable progress has been made in this respect by the genetic modification of pigs to protect their tissues from the primate immune response and to correct several molecular incompatibilities that exist between pig and primate. These have included knockout of genes responsible for the expression of major antigenic targets for primate natural anti-pig antibodies, insertion of human complement- and coagulation-regulatory transgenes, and knockdown of swine leukocyte antigens that stimulate the primate＇s adaptive immune response. As a result of these manipulations, the administration of novel immunosuppressive agents, and other innovations, pig hearts have now functioned in baboons for 6-8 months, pig islets have maintained normoglycemia in diabetic monkeys for 〉 1 year, and pig corneas have maintained transparency for several months. Clinical trials of pig islet trans- plantation are already in progress. Future developments will involve further genetic manipulations of the organ- source pig, with most of the genes that are likely to be beneficial already identified.
基金the Russian Science Foundation(No.16-15-00010to RRI)the Russian Government Program of Competitive Growth of Kazan Federal University。
文摘We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy for neurodegenerative diseases,neurotrauma,and stroke.In this study,using a mini pig model of spinal cord injury,we proposed for the first time the use of gene-modified leucoconcentrate prepared from peripheral blood in the plastic blood bag for personalized ex vivo gene therapy.Leucoconcentrate obtained from mini pig peripheral blood was transduced with a chimeric adenoviral vector(Ad5/35 F)that carried an enhanced green fluorescent protein(EGFP)reporter gene in the plastic blood bag.The day after blood donation,the mini pigs were subjected to moderate SCI and four hours post-surgery they were intravenously autoinfused with gene-modified leucoconcentrate.A week after gene-modified leucoconcentrate therapy,fluorescent microscopy revealed EGFP-expressing leucocytes in spinal cord at the site of contusion injury.In the spleen the groups of EGFP-positive cells located in the lymphoid follicles were observed.In vitro flow cytometry and fluorescent microscopy studies of the gene-modified leucoconcentrate samples also confirmed the production of EGFP by leucocytes.Thus,the efficacy of leucocytes transduction in the plastic blood bag and their migratory potential suggest their use for temporary production of recombinant biologically active molecules to correct certain pathological conditions.This paper presents a proof-of-concept of simple,safe and effective approach for personalized ex vivo gene therapy based on gene-modified leucoconcentrate autoinfusion.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.5)on May 27,2014.
