Objective: In this paper we compared the two methods of cell sorting (magnetic cell sorting and flow cytometry sorting) for the isolation and function analysis of mouse CD4+ CD25+ regulatory T (Treg) cells, in order t...Objective: In this paper we compared the two methods of cell sorting (magnetic cell sorting and flow cytometry sorting) for the isolation and function analysis of mouse CD4+ CD25+ regulatory T (Treg) cells, in order to inform further studies in Treg cell function. Methods: We separately used magnetic cell sorting and flow cytometry sorting to identify CD4+ CD25+ Treg cells. After magnetic cell separation, we further used flow cytometry to analyze the purity of CD4+ CD25+ Treg cells, trypan blue staining to detect cell viability, and propidium iodide (PI) staining to assess the cell viability. We detected the immune inhibition of CD4+ CD25+ Treg cells in the in vitro proliferation experiments. Results: The results showed that compared to flow cytometry sorting, magnetic cell sorting took more time and effort, but fewer live cells were obtained than with flow cytometry sorting. The CD4+ CD25+ Treg cells, however, obtained with both methods have similar immunosuppressive capacities. Conclusion: The result suggests that both methods can be used in isolating CD4+ CD25+ Treg cells, and one can select the best method according to specific needs and availability of the methodologies.展开更多
Objective： To investigate the effects of diltiazem and cyclosporine A （CsA） combination therapy on protecting the kidney, promoting graft functioning and improving post-transplanted kidney recovery. Methods： The b...Objective： To investigate the effects of diltiazem and cyclosporine A （CsA） combination therapy on protecting the kidney, promoting graft functioning and improving post-transplanted kidney recovery. Methods： The blood con- centrations of CsA, the condition of the post-transplant kidney, the rate of acute rejection （AR）, as well as hepatic and renal toxicity in 636 cases of renal transplant recipients were determined after being treated by CsA, with or without diltiazem. Results： Compared with the control group which received CsA, mycophenolate mofetil （MMF） and prednisolone （Pred） but lacked diltiazem, the group receiving these agents together with diltiazem required reduced dosage of CsA （P 〈 0.01）, while blood concentrations of CsA were significantly increased （P 〈 0.01）; the recovery time of graft function was reduced from （6.2± 1.5） d to （3.9± 1.4） d （P 〈 0.01）, and the rate of AR was decreased from 13.2% to 7.9% （P 〈 0.01）. Conclusion： In renal transplantation patients treated with CsA and diltiazem, blood concentrations of CsA were increased while the dosage was decreased. This efficient combination therapy reduced patients economic burden, at the same time retained kidney function, promoted graft function recovery and decreased hepatic and renal toxicity and the rate of AR.展开更多
The feasibility and the clinical value of the enzyme-multiplied immunoassay technique （EMIT） monitoring of blood concentrations of cyclosporine A （CsA） in patients treated with CsA were investigated after kidney t...The feasibility and the clinical value of the enzyme-multiplied immunoassay technique （EMIT） monitoring of blood concentrations of cyclosporine A （CsA） in patients treated with CsA were investigated after kidney transplantation. The validation method was performed to the EMIT determination of CsA blood concentration, the CsA whole blood trough concentrations （Co） of patients in different time periods after renal transplantation were monitored, and combined with the clinical complications, the statistical results were analyzed and compared. EMIT was precise, accurate and stable, also with a high quality control. The mean postoperative blood concentration of CsA was as follows： 〈1 month, （281.4± 57.9）ng/mL; 2 - 3 months, （264.5 ± 41.2） ng/mL; 4 - 5 months, （236.4 ± 38.9） ng/mL; 6 - 12 months, （206.5± 32.6）ng/mL; 〉12 months, （185.6± 28.1）ng/mL. The toxic reaction rate of CsA blood concentration within the recommended therapeutic concentration was 14.1%, significantly lower than that of the none-recommended dose group （37.2%） （P〈0.05）; the transplantation rejection rate was 4.4%, significantly lower than that of the none- recommended dose group （22.5%） （P〈0.05）. Using EMIT to monitor the blood concentration of CsA as the routine laboratory method is feasible, and is able to reduce the CsA toxicity and rejection significantly, leading to achieving the desired therapeutic effect.展开更多
Objective： To explore the effects of cytomegalovirus （CMV） infection on rejection-related gene expression in the endothelial cells of renal transplantation recipients. Methods： Endothelial cells （ECs） were cultu...Objective： To explore the effects of cytomegalovirus （CMV） infection on rejection-related gene expression in the endothelial cells of renal transplantation recipients. Methods： Endothelial cells （ECs） were cultured and stimulated by a variety of factors： A, normal control group; B, inactivated human cytomegalovirus （HCMV） infection group; C, HCMV infection group; D, HCMV supematant infection group; and E, ganciclovir HCMV group. Expression of intercellular adhesion molecule-1 （ICAM-1） and major histocompability complex （MHC） class I and class II antigens was detected by flow cytometry （FCM） and immuno- histochemistry. Results： We found characteristic CMV-infected ECs in this study. There were no significant differences among groups A, B and D （P〉0.05）. Although the expression levels of ICAM-1 were not significantly different between groups C and E （P〉0.05）, the ICAM-1 expression in these two groups was significantly higher than that in group A （P〈0.05）. ICAM-1 expression was detected in groups C and E, while there was no expression in groups A, B and D. Furthermore, there was no significant difference of ICAM-1 mRNA expression between groups C and E （P〉0.05）. Human leucocyte antigen （HLA）-ABC expression was detected in all the groups, while HLA-DR expression was only detected in groups C and E. There were no significant dif- ferences of HLA-ABC and HLA-DR expression among groups A, B and D （P〉0.05）. However, the HLA-ABC and HLA-DR expression levels in groups C and D were higher than those of the remaining groups previously reported （P〈0.05）. Meanwhile, the HLA-ABC and HLA-DR expression levels in group E were lower than those of group C （P〈0.05）. Conclusion： CMV could up-regulate the expression levels of ICAM-1 and MHC antigens, which was closely related to allograft rejection.展开更多
Objective To investigate the effect of prostaglandin E1 (PGE1) on recovery of early renal graft functions after transplantation. Methods One hundred and seven patients after renal transplantation were allocated in the...Objective To investigate the effect of prostaglandin E1 (PGE1) on recovery of early renal graft functions after transplantation. Methods One hundred and seven patients after renal transplantation were allocated in the treated group, and treated by conventional treatment with injection of 10 μg prostaglandin E1 additionally twice a day for 14 days. And eighty-eight patients who received conventional treatment alone after renal transplantation at the corresponding period were allocated in the control group. Indexes of the two groups, including incidence of delayed graft function and acute rejection reaction, volume of urine, serum certaintie (SCr), endogenous certainties clearance rate (CCr), the blood flow resistance in graft as well as blood viscosity (BV), and platelet aggregation rate (PAR), were determined. Results The urinary volume and endogenous certainties clearance rate of the treated group were significantly higher, but the level of SCr, incidence of renal function recovery retardation, BV, PAR and blood flow resistance in graft were significantly lower than those of the control group (P<0.05). The difference of incidence of acute rejection reaction between the two groups was insignificant (P>0.05). Conclusion Prostaglandin E1 can improve blood microcirculation and decrease the incidence of renal function recovery retardation. These effects are helpful for recovery of renal function after renal transplantation.展开更多
Background:Kidney transplant is always emergent operations and frequently need to be performed at nighttime to reduce cold ischemia time(CIT).Previous studies have revealed that fatigue and sleep deprivation can resul...Background:Kidney transplant is always emergent operations and frequently need to be performed at nighttime to reduce cold ischemia time(CIT).Previous studies have revealed that fatigue and sleep deprivation can result in adverse consequences of medical procedures.This study aimed to evaluate whether nighttime operation has adverse impact on kidney transplant.Methods:A retrospective analysis of recipients accepted kidney transplant from deceased donors in one center from 2014 to 2016 was performed.Daytime transplant was defined as operation started after 8 AM or ended before 8 PM and nighttime operation was defined as operation ended after 8 PM or started before 8 AM.The incidences of complications such as delayed graft function,acute rejection,surgical complications and nosocomial infections were compared between 2 groups.Student’s t-test was used to analyze continuous variables such as serum creatinine(Scr)at 1-year of post-transplant.The Chi-square test was used to analyze categorical variables.Differences in recipients and graft survival were analyzed using Kaplan–Meier methodology and log-rank tests.Results:Among the 443 recipients,233(52.6%)were classified into the daytime group and the others 210(47.4%)were in the nighttime group.The 1-year survival rate of recipients was similar for the recipients in the daytime and nighttime groups(95.3%vs.95.2%,P=0.981).Although the 1-year graft survival rate in the nighttime group was slightly superior to that in the daytime group,the difference was not significant(92.4%vs.88.4%,P=0.164).Furthermore,Scr and incidence of complications were also not significantly different between the 2 groups.Conclusions:Our results suggested that operation time of kidney transplant with short CIT has no significant impact on the outcome of kidney transplant.Nighttime operation of kidney transplant with short CIT could be postponed to the following day to alleviate the burden on medical staffs and avoid the potential risk.展开更多
Objective： To evaluate the efficacy of kushenin in treating patients with chronic hepatitis C after renal transplantation. Methods： Fifty-five patients were randomly assigned by lottery to the treatment group （29 c...Objective： To evaluate the efficacy of kushenin in treating patients with chronic hepatitis C after renal transplantation. Methods： Fifty-five patients were randomly assigned by lottery to the treatment group （29 cases） and control group （26 cases）. The same immunosuppression therapy was given to all patients in both groups. Patients in the treatment group were treated with kushenin 0.6 g once a day, while those in the control group were treated with conventional liver protective agents such as vitamins. The treatment duration of both groups was 3 months. The incidences of serious hepatitis and acute rejection reaction, serum biochemistry parameters including indicators of liver and kidney functions, hepatic fibrosis index, and serum HCV-RNA were compared between the two groups. Results： （1） The incidence of serious hepatitis in the treatment group and the control group was 3.45% （1/29 cases） and 11.54% （3/26 cases）, respectively, which was insignificantly different between the two groups （P=0.335）. （2） The incidence of acute rejection in the treatment group was 6.90% （2/29 cases） and that in the control group was 7.69% （2/26 cases）, showing insignificant difference （P=0.335）. （3） The differences in serum alanine aminotransferase （ALT）, direct bilirubin （DBIL）, hyaluronic acid （HA）, propeptide collagen type Ⅲ （PC Ⅲ）, laminin （LN）, collagen type Ⅳ（Col Ⅳ） levels between the two groups were insignificant before transplantation （P〉0.05）, while the above-mentioned parameters in the treatment group were significantly lower than those in the control group after transplantation （P〈0.05）. The difference in serum creatinine （SCr） and endogenous creatinine clearance rate （CCr） between the two groups was insignificant before and after transplantation （P〉0.05）. （4） The negative conversion rate of HCV-RNA in the treatment group was 31.03% （9/29 cases）, significantly higher than the value of 11.54% （3/26 cases） in the control group after transplantation （P〈0.05）. （5） The levels of serum ALT and DBIL in patients with HCV-RNA converted to negative were significantly lower than those with still-positive HCV-RNA （P〈0.05）. Conclusions： Kushenin has a certain effect on inhibiting the proliferation of HCV, protecting liver cells, and anti-liver fibrosis. On the other hand, it has no obvious influence on renal allograft function. Thus, the drug is clinically safe and effective for use in treating patients with chronic hepatitis C after renal transplantation.展开更多
Cholesterol crystal embolism(CCE),characterized by diverse clinical presentation,limited therapeutic options and poor long-term outcomes,is a multi-systemic disease in which cholesterol crystals emboli released from a...Cholesterol crystal embolism(CCE),characterized by diverse clinical presentation,limited therapeutic options and poor long-term outcomes,is a multi-systemic disease in which cholesterol crystals emboli released from atherosclerotic plaques of the aorta or other major arteries occlude small arteries and subsequently lead to a foreign-body giant cell granuloma inflammatory reaction.1 Invasive vascular procedure.展开更多
基金Project supported by the National Natural Science Foundation of China (Nos. 30872578 and 30753761)the Natural Science Founda-tion of Shanxi Province (No. SJ08C201)+1 种基金the Science and Technology Key Projects Foundation of Shanxi Province (No. 2008K13-04)the Science and Technology Plan Projects Foundation of Xi’an (No. SF08006-2), China
文摘Objective: In this paper we compared the two methods of cell sorting (magnetic cell sorting and flow cytometry sorting) for the isolation and function analysis of mouse CD4+ CD25+ regulatory T (Treg) cells, in order to inform further studies in Treg cell function. Methods: We separately used magnetic cell sorting and flow cytometry sorting to identify CD4+ CD25+ Treg cells. After magnetic cell separation, we further used flow cytometry to analyze the purity of CD4+ CD25+ Treg cells, trypan blue staining to detect cell viability, and propidium iodide (PI) staining to assess the cell viability. We detected the immune inhibition of CD4+ CD25+ Treg cells in the in vitro proliferation experiments. Results: The results showed that compared to flow cytometry sorting, magnetic cell sorting took more time and effort, but fewer live cells were obtained than with flow cytometry sorting. The CD4+ CD25+ Treg cells, however, obtained with both methods have similar immunosuppressive capacities. Conclusion: The result suggests that both methods can be used in isolating CD4+ CD25+ Treg cells, and one can select the best method according to specific needs and availability of the methodologies.
基金supported by the"13115"Innovation Technology Project of Special Purpose of Shaanxi Province(No.2087ZDKG-67)the National Natural Science Foundation of China(No.30772096)+2 种基金the Natural Science Foundation of Shaanxi Province(No.2007C2C12)the Project of the National Science Foundation for Distinguished Young Scholars of First Affiliated Hospital of the Medical College of Xi'an Jiaotong University(No.2006YK4)the Science Research of Sha-anxi Health Department(No.08D25)
文摘Objective： To investigate the effects of diltiazem and cyclosporine A （CsA） combination therapy on protecting the kidney, promoting graft functioning and improving post-transplanted kidney recovery. Methods： The blood con- centrations of CsA, the condition of the post-transplant kidney, the rate of acute rejection （AR）, as well as hepatic and renal toxicity in 636 cases of renal transplant recipients were determined after being treated by CsA, with or without diltiazem. Results： Compared with the control group which received CsA, mycophenolate mofetil （MMF） and prednisolone （Pred） but lacked diltiazem, the group receiving these agents together with diltiazem required reduced dosage of CsA （P 〈 0.01）, while blood concentrations of CsA were significantly increased （P 〈 0.01）; the recovery time of graft function was reduced from （6.2± 1.5） d to （3.9± 1.4） d （P 〈 0.01）, and the rate of AR was decreased from 13.2% to 7.9% （P 〈 0.01）. Conclusion： In renal transplantation patients treated with CsA and diltiazem, blood concentrations of CsA were increased while the dosage was decreased. This efficient combination therapy reduced patients economic burden, at the same time retained kidney function, promoted graft function recovery and decreased hepatic and renal toxicity and the rate of AR.
基金supported by the Project 973 Monitoring of the Immune Status and Rejection After Organ Transplantation"(2009CB522400)the National Natural Science Foundation of China(No.30972947)
文摘The feasibility and the clinical value of the enzyme-multiplied immunoassay technique （EMIT） monitoring of blood concentrations of cyclosporine A （CsA） in patients treated with CsA were investigated after kidney transplantation. The validation method was performed to the EMIT determination of CsA blood concentration, the CsA whole blood trough concentrations （Co） of patients in different time periods after renal transplantation were monitored, and combined with the clinical complications, the statistical results were analyzed and compared. EMIT was precise, accurate and stable, also with a high quality control. The mean postoperative blood concentration of CsA was as follows： 〈1 month, （281.4± 57.9）ng/mL; 2 - 3 months, （264.5 ± 41.2） ng/mL; 4 - 5 months, （236.4 ± 38.9） ng/mL; 6 - 12 months, （206.5± 32.6）ng/mL; 〉12 months, （185.6± 28.1）ng/mL. The toxic reaction rate of CsA blood concentration within the recommended therapeutic concentration was 14.1%, significantly lower than that of the none-recommended dose group （37.2%） （P〈0.05）; the transplantation rejection rate was 4.4%, significantly lower than that of the none- recommended dose group （22.5%） （P〈0.05）. Using EMIT to monitor the blood concentration of CsA as the routine laboratory method is feasible, and is able to reduce the CsA toxicity and rejection significantly, leading to achieving the desired therapeutic effect.
基金Project supported by the National Natural Science Foundation of China (No. 30772096)the Clinical Key Disciplines of National Public Health Department, the Major Scientific and Technological Special Projects of Shannxi Province (No. 2007ZDKG-67)the Natural Science Foundation of Shaanxi Province (No. 30571799), China
文摘Objective： To explore the effects of cytomegalovirus （CMV） infection on rejection-related gene expression in the endothelial cells of renal transplantation recipients. Methods： Endothelial cells （ECs） were cultured and stimulated by a variety of factors： A, normal control group; B, inactivated human cytomegalovirus （HCMV） infection group; C, HCMV infection group; D, HCMV supematant infection group; and E, ganciclovir HCMV group. Expression of intercellular adhesion molecule-1 （ICAM-1） and major histocompability complex （MHC） class I and class II antigens was detected by flow cytometry （FCM） and immuno- histochemistry. Results： We found characteristic CMV-infected ECs in this study. There were no significant differences among groups A, B and D （P〉0.05）. Although the expression levels of ICAM-1 were not significantly different between groups C and E （P〉0.05）, the ICAM-1 expression in these two groups was significantly higher than that in group A （P〈0.05）. ICAM-1 expression was detected in groups C and E, while there was no expression in groups A, B and D. Furthermore, there was no significant difference of ICAM-1 mRNA expression between groups C and E （P〉0.05）. Human leucocyte antigen （HLA）-ABC expression was detected in all the groups, while HLA-DR expression was only detected in groups C and E. There were no significant dif- ferences of HLA-ABC and HLA-DR expression among groups A, B and D （P〉0.05）. However, the HLA-ABC and HLA-DR expression levels in groups C and D were higher than those of the remaining groups previously reported （P〈0.05）. Meanwhile, the HLA-ABC and HLA-DR expression levels in group E were lower than those of group C （P〈0.05）. Conclusion： CMV could up-regulate the expression levels of ICAM-1 and MHC antigens, which was closely related to allograft rejection.
基金supported by the National Natural Science Foundation of China (No.30471640)
文摘Objective To investigate the effect of prostaglandin E1 (PGE1) on recovery of early renal graft functions after transplantation. Methods One hundred and seven patients after renal transplantation were allocated in the treated group, and treated by conventional treatment with injection of 10 μg prostaglandin E1 additionally twice a day for 14 days. And eighty-eight patients who received conventional treatment alone after renal transplantation at the corresponding period were allocated in the control group. Indexes of the two groups, including incidence of delayed graft function and acute rejection reaction, volume of urine, serum certaintie (SCr), endogenous certainties clearance rate (CCr), the blood flow resistance in graft as well as blood viscosity (BV), and platelet aggregation rate (PAR), were determined. Results The urinary volume and endogenous certainties clearance rate of the treated group were significantly higher, but the level of SCr, incidence of renal function recovery retardation, BV, PAR and blood flow resistance in graft were significantly lower than those of the control group (P<0.05). The difference of incidence of acute rejection reaction between the two groups was insignificant (P>0.05). Conclusion Prostaglandin E1 can improve blood microcirculation and decrease the incidence of renal function recovery retardation. These effects are helpful for recovery of renal function after renal transplantation.
文摘Background:Kidney transplant is always emergent operations and frequently need to be performed at nighttime to reduce cold ischemia time(CIT).Previous studies have revealed that fatigue and sleep deprivation can result in adverse consequences of medical procedures.This study aimed to evaluate whether nighttime operation has adverse impact on kidney transplant.Methods:A retrospective analysis of recipients accepted kidney transplant from deceased donors in one center from 2014 to 2016 was performed.Daytime transplant was defined as operation started after 8 AM or ended before 8 PM and nighttime operation was defined as operation ended after 8 PM or started before 8 AM.The incidences of complications such as delayed graft function,acute rejection,surgical complications and nosocomial infections were compared between 2 groups.Student’s t-test was used to analyze continuous variables such as serum creatinine(Scr)at 1-year of post-transplant.The Chi-square test was used to analyze categorical variables.Differences in recipients and graft survival were analyzed using Kaplan–Meier methodology and log-rank tests.Results:Among the 443 recipients,233(52.6%)were classified into the daytime group and the others 210(47.4%)were in the nighttime group.The 1-year survival rate of recipients was similar for the recipients in the daytime and nighttime groups(95.3%vs.95.2%,P=0.981).Although the 1-year graft survival rate in the nighttime group was slightly superior to that in the daytime group,the difference was not significant(92.4%vs.88.4%,P=0.164).Furthermore,Scr and incidence of complications were also not significantly different between the 2 groups.Conclusions:Our results suggested that operation time of kidney transplant with short CIT has no significant impact on the outcome of kidney transplant.Nighttime operation of kidney transplant with short CIT could be postponed to the following day to alleviate the burden on medical staffs and avoid the potential risk.
基金National Natural Science Foundation of China(No.30571799,30471640)
文摘Objective： To evaluate the efficacy of kushenin in treating patients with chronic hepatitis C after renal transplantation. Methods： Fifty-five patients were randomly assigned by lottery to the treatment group （29 cases） and control group （26 cases）. The same immunosuppression therapy was given to all patients in both groups. Patients in the treatment group were treated with kushenin 0.6 g once a day, while those in the control group were treated with conventional liver protective agents such as vitamins. The treatment duration of both groups was 3 months. The incidences of serious hepatitis and acute rejection reaction, serum biochemistry parameters including indicators of liver and kidney functions, hepatic fibrosis index, and serum HCV-RNA were compared between the two groups. Results： （1） The incidence of serious hepatitis in the treatment group and the control group was 3.45% （1/29 cases） and 11.54% （3/26 cases）, respectively, which was insignificantly different between the two groups （P=0.335）. （2） The incidence of acute rejection in the treatment group was 6.90% （2/29 cases） and that in the control group was 7.69% （2/26 cases）, showing insignificant difference （P=0.335）. （3） The differences in serum alanine aminotransferase （ALT）, direct bilirubin （DBIL）, hyaluronic acid （HA）, propeptide collagen type Ⅲ （PC Ⅲ）, laminin （LN）, collagen type Ⅳ（Col Ⅳ） levels between the two groups were insignificant before transplantation （P〉0.05）, while the above-mentioned parameters in the treatment group were significantly lower than those in the control group after transplantation （P〈0.05）. The difference in serum creatinine （SCr） and endogenous creatinine clearance rate （CCr） between the two groups was insignificant before and after transplantation （P〉0.05）. （4） The negative conversion rate of HCV-RNA in the treatment group was 31.03% （9/29 cases）, significantly higher than the value of 11.54% （3/26 cases） in the control group after transplantation （P〈0.05）. （5） The levels of serum ALT and DBIL in patients with HCV-RNA converted to negative were significantly lower than those with still-positive HCV-RNA （P〈0.05）. Conclusions： Kushenin has a certain effect on inhibiting the proliferation of HCV, protecting liver cells, and anti-liver fibrosis. On the other hand, it has no obvious influence on renal allograft function. Thus, the drug is clinically safe and effective for use in treating patients with chronic hepatitis C after renal transplantation.
文摘Cholesterol crystal embolism(CCE),characterized by diverse clinical presentation,limited therapeutic options and poor long-term outcomes,is a multi-systemic disease in which cholesterol crystals emboli released from atherosclerotic plaques of the aorta or other major arteries occlude small arteries and subsequently lead to a foreign-body giant cell granuloma inflammatory reaction.1 Invasive vascular procedure.
