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Background@#Vascular resistance and flow rate during hypothermic machine perfusion (HMP) of kidneys is correlated with graft function. We aimed to determine the effects of increasing HMP pressure versus maintaining the initial pressure on kidney transplantation outcomes.@*Methods@#We retrospectively reviewed the data of 76 primary transplantation patients who received HMP-preserved kidneys from 48 donors after cardiac death between September 1, 2013, and August 31, 2015. HMP pressure was increased from 30 to 40 mmHg (1 mmHg = 0.133 kPa) in kidneys with poor flow and/or vascular resistance (increased pressure [IP] group; 36 patients); otherwise, the initial pressure was maintained (constant pressure group; 40 patients). Finally, the clinical characteristics and transplantation outcomes in both groups were assessed.@*Results@#Delayed graft function (DGF) incidence, 1-year allograft, patient survival, kidney function recovery time, and serum creatinine level on day 30 were similar in both groups, with improved flow and resistance in the IP group. Among patients with DGF, kidney function recovery time and DGF duration were ameliorated in the IP group. Multivariate logistic regression analysis revealed that donor hypertension (odds ratio [OR]: 1.43, 95% confidence interval [CI]: 1.02-2.06, P = 0.035), donor terminal serum creatinine (OR: 1.27, 95% CI: 1.06-1.62, P = 0.023), warm ischemic time (OR: 3.45, 95% CI: 1.97-6.37, P = 0.002), and terminal resistance (OR: 3.12, 95% CI: 1.76-6.09, P = 0.012) were independent predictors of DGF. Cox proportional hazards analysis showed that terminal resistance (hazard ratio: 2.06, 95% CI: 1.32-5.16, P = 0.032) significantly affected graft survival.@*Conclusion@#Increased HMP pressure improves graft perfusion but does not affect DGF incidence or 1-year graft survival.
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Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Aloinjertos , Funcionamiento Retardado del Injerto , Hipertensión , Pruebas de Función Renal , Trasplante de Riñón , Métodos , Modelos Logísticos , Preservación de Órganos , Estudios Retrospectivos , Donantes de TejidosRESUMEN
Objective To investigate the role of RUNX3 in the regulation of macrophage polarization so as to provide a new therapeutic approach for immunity-related diseases.Methods ① After RAW264.7 cells were stimulated by IFN-γ,LPS and IL-4,respectively,the expressions of their surface markers(arginase-1 and iNOS) were detected by RT-PCR to observe whether RAW264.7 cells polarized to M1 or M2 after stimulation by IFN-γ, LPS and IL-4.The cells stimulated by IFN-γ and LPS were named group M1 and those stimulated by IL-4 were group M2;the control group was group M0.② The expression of RUNX3 was detected by immunofluorescence and RT-PCR methods in each cell group(M1,M2 and M0).③ RUNX3 over-expression vector was established.The RUNX3 gene in RAW264.7 cells was silenced.Cell lines with stable expression were screened with G 418 culture medium.The expressions of cell surface markers iNOS and CD86 were detected by RT-PCR;cell secretion(TNF-α) was detected using ELESA method.Results ① Stimulation of RAW264.7 cells with IFN-γ and LPS could induce RAW264.7 cells to polarize into M1 type macrophages.The mRNA expression of iNOS in M1 group was higher than that in group M0 detected by RT-PCR(P= 0.002),while using IL-4 to stimulate RAW264.7 cells could induce RAW264.7 cells to polarize into M2 macrophages.The results of RT-PCR detection showed that the expression of arginase-1 was higher in M2 group than in group M0(P=0.021).② The expression of RUNX3 mRNA in the M1 cells group was higher than that in the M0 cells group(P= 0.001),but the expression in the M2 cells group was decreased(P=0.041).③ After silencing of RUNX3,the expressions of RAW264.7 cell surface markers CD86 and iNOS(P=0.005)and the cells secretion of TNF-α(P<0.001)were decreased compared with those in the control group.Conclusion RUNX3 transcriptional activation may promote the differentiation of macrophages into M1 type.
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<p><b>Background</b>Immunosuppressive agents are still inefficient in preventing biopsy-proven acute rejection (BPAR) after expanded criteria donor (ECD) kidney transplantation. The aim of this study was to investigate the relationships between early immunosuppressive exposure and the development of BPAR.</p><p><b>Methods</b>We performed a retrospective study of 58 recipients of ECD kidney transplantation treated with enteric-coated-mycophenolate sodium, tacrolimus (Tac), and prednisone. The levels of mycophenolic acid-area under the curve (MPA-AUC) and Tac Cwere measured at the 1 week and the 1 month posttransplant, respectively. The correlation was assessed by multivariate logistic regression.</p><p><b>Results</b>The occurrence rates of BPAR and antibody-mediated rejection were 24.1% and 10.3%, respectively. A low level of MPA-AUC at the 1 week posttransplant was found in BPAR recipients (38.42 ± 8.37 vs. 50.64 ± 13.22, P < 0.01). In addition, the incidence of BPAR was significantly high (P < 0.05) when the MPA-AUClevel was <30 mg·h·L at the 1 week (15.0% vs. 44.4%) or the Tac Cwas <4 ng/ml at the 1 month posttransplant (33.3% vs. 21.6%). Multivariable logistic regression analysis showed that the MPA-AUC at the 1 week (OR: 0.842, 95% CI: 0.784-0.903) and the Tac Cat the 1 month (OR: 0.904, 95% CI: 0.822-0.986) had significant inverse correlation with BPAR (P < 0.05).</p><p><b>Conclusions</b>Low-level exposure of MPA and Tac Cin the early weeks posttransplant reflects an increased acute rejection risk, which suggested that MPA-AUC <30 mg·h·L and Tac C <4 ng/ml should be avoided in the first few weeks after transplantation.</p>
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Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Rechazo de Injerto , Alergia e Inmunología , Inmunosupresores , Química , Usos Terapéuticos , Trasplante de Riñón , Métodos , Ácido Micofenólico , Química , Usos Terapéuticos , Estudios Retrospectivos , Tacrolimus , Química , Usos Terapéuticos , Factores de TiempoRESUMEN
<p><b>BACKGROUND</b>Improving islet graft revascularization has become a crucial task for prolonging islet graft survival. Endothelial cells (ECs) are the basis of new microvessels in an isolated islet, and EC coating has been demonstrated to improve the vascularization and survival of an islet. However, the traditional method of EC coating of islets has low efficiency in vitro. This study was conducted to evaluate the effect of a polyglycolic acid (PGA) scaffold on the efficiency of islet coating by ECs and the angiogenesis in the coated islet graft.</p><p><b>METHODS</b>A PGA fibrous scaffold was used for EC coating of islet culture and was evaluated for its efficiency of EC coating on islets and islet graft angiogenesis.</p><p><b>RESULTS</b>In in vitro experiments, we found that apoptosis index of ECs-coating islet in PGA group (27% ± 8%) was significantly lower than that in control group (83% ± 20%, P < 0.05) after 7 days culture. Stimulation index was significantly greater in the PGA group than in the control group at day 7 after ECs-coating (2.07 ± 0.31 vs. 1.80 ± 0.23, P < 0.05). vascular endothelial growth factor (VEGF) level in the PGA group was significantly higher than the coating in the control group after 7 days culture (52.10 ± 13.50 ng/ml vs. 16.30 ± 8.10 ng/ml, P < 0.05). Because of a tight, circumvallated, adhesive and three-dimensional growth microenvironment, islet cultured in a PGA scaffold had higher coating efficiency showing stronger staining intensity of enzyme than those in the control group after 14 days of culture following ECs-coating. For in vivo study, PGA scaffold significantly prolonged the average survival time of EC-coated islet graft after transplantation compared with control group (15.30 ± 5.60 days vs. 8.30 ± 2.45 days, P < 0.05). The angiogenesis and area of survived grafts were more in the PGA group compared with the control group by measuring the mean microvessel density (8.60 ± 1.21/mm2 vs. 5.20 ± 0.87/mm2, P < 0.05). In addition, expression of VEGF and tyrosin-protein kinase receptor (Tie-2) gene increased in PGA scaffold group than that in control group by real-time reverse transcription-polymerase chain reaction analysis.</p><p><b>CONCLUSIONS</b>These results demonstrate that the efficiency of EC coating of islets was successfully increased by culturing ECs on a PGA scaffold. This method enhances the function, survival, and vascularization of isolated islets in vitro and in vivo.</p>
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Animales , Ratas , Apoptosis , Células Endoteliales , Ensayo de Inmunoadsorción Enzimática , Supervivencia de Injerto , Insulina , Metabolismo , Islotes Pancreáticos , Trasplante de Islotes Pancreáticos , Métodos , Neovascularización Fisiológica , Ácido Poliglicólico , Química , Farmacología , Ratas Sprague-Dawley , Ratas Wistar , Andamios del Tejido , QuímicaRESUMEN
<p><b>BACKGROUND</b>How to evaluate the quality of donation after cardiac death (DCD) kidneys has become a critical problem in kidney transplantation in China. Hence, the aim of this study was to develop a simple donor risk score model to evaluate the quality of DCD kidneys before DCD.</p><p><b>METHODS</b>A total of 543 qualified kidneys were randomized in a 2:1 manner to create the development and validation cohorts. The donor variables in the development cohort were considered as candidate univariate predictors of delayed graft function (DGF). Multivariate logistic regression was then used to identify independent predictors of DGF with P < 0.05. Date from validation cohort were used to validate the donor scoring model.</p><p><b>RESULTS</b>Based on the odds ratios, eight identified variables were assigned a weighted integer; the sum of the integer was the total risk score for each kidney. The donor risk score, ranging from 0 to 28, demonstrated good discriminative power with a C-statistic of 0.790. Similar results were obtained from validation cohort with C-statistic of 0.783. Based on the obtained frequencies of DGF in relation to different risk scores, we formed four risk categories of increasing severity (scores 0-4, 5-9, 10-14, and 15-28).</p><p><b>CONCLUSIONS</b>The scoring model might be a good noninvasive tool for assessing the quality of DCD kidneys before donation and potentially useful for physicians to make optimal decisions about donor organ offers.</p>
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<p><b>OBJECTIVE</b>To study the effect of Poria cocos (Pcs) in preventing acute rejection of rats after renal transplantation and its mechanism.</p><p><b>METHODS</b>Rat orthotopic renal transplantation model was performed with Wistar rat as donor and SD rat as donee. All donees were divided into 4 groups, 10 in each group, before transplantation. They were treated respectively with normal saline 5 mL x kg(-1) x d(-1) (A), Pcs 25 mg x kg(-1) x d(-1) (B), Pcs 50 mg x kg(-1) x d(-1) (C) and ciclosporin A (CsA) 5 mg x kg(-1) x d(-1) (D) by intragastric administration. The renal allograft survival time (ST) was recorded, and the serum levels of creatinine (SCr), interleukin-2 (IL-2), gamma-interferon (gamma-IFN), CD4+, CD8+ lymphocytes percentage, CD4+/CD8+ ratio, as well as the pathologic changes were observed one week after transplantation.</p><p><b>RESULTS</b>ST of the renal graft in Groups C and D was significantly longer with pathologic change evidently less than those in Groups A and B (P<0.01), and the ST in Group C was shorter that in Group D (P<0.05). Changes of renal function and urine volume were identified to the pathological change of graft, the initiating time of renal dysfunction was later in Groups C and D than that in Groups A and B. Serum levels of IL-2, IFN-gamma and CD4+ percentage in Group C were significantly lower than those in Groups A and B, but higher than those in Group D respectively (P<0.05 or P<0.01), while CD8+ percentage in Group C was significantly lower than that in Group A (P<0.05), but insignificantly different to that in Groups B and D (P>0.05).</p><p><b>CONCLUSION</b>Pcs shows good dosage-dependent effect in suppressing acute rejection of renal transplantation, but the effect is inferior to that of CsA.</p>
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Animales , Masculino , Ratas , Ciclosporina , Usos Terapéuticos , Rechazo de Injerto , Trasplante de Riñón , Materia Medica , Usos Terapéuticos , Poria , Química , Ratas Sprague-Dawley , Ratas WistarRESUMEN
<p><b>OBJECTIVE</b>To retrospectively study and analyse the immune regulatory effect of Bailing Capsule (BLC, a dry powder preparation of Cordyceps sinensis mycelia) on patients after renal transplantation, its influences on various systems of organism, and to explore its possible acting mechanism.</p><p><b>METHODS</b>In accordance with the entry criteria, 67 recipients of renal homo-allograft were assigned to two groups. The 42 cases in the control group were treated with mycophenolate mofetil (MMF) plus cyclosporine A (CsA), or tacrolimus (FK506) plus prednisone (Pred); the 25 in the treated group treated with the chemotherapy the same as in the control group plus BLC. They were followed up for 48 weeks by checking up blood routine, urine routine, hepatic and renal function, total serum protein, serum albumin, uric acid, etc., and the dosage of immunoinhibitory used was recorded periodically.</p><p><b>RESULTS</b>Comparison showed no significant difference in graft survival rate, occurrence of reject reaction and renal function recovery between the two groups; but levels of urinary erythrocytes and leucocytes, blood alanine transaminase, aspartate amino transferase, uric acid, total bilirubin, direct bilirubin, as well as the incidence of infection were significantly lower, and serum total protein and albumin were significantly higher in the treated group (all P < 0.01); moreover, counts of erythrocyte and leukocyte from 12 to 48 weeks, T-lymphocyte from 4 to 48 weeks after transplantation were significantly higher in the treated group (P < 0.05 and P < 0.01), and the recovery appeared earlier, the dosage of CsA or FK506 used 12 weeks after operation was significantly lower in the treated group than in the control group (P < 0.05, P < 0.01).</p><p><b>CONCLUSIONS</b>BLC could effectively protect liver and kidney, stimulate hemopoietic function, improve hypoproteinemia, as well as reduce the incidence of infection and the dosage of CsA and FK506 used, etc. Therefore, it is a useful drug for immunoregulation after organ transplantation.</p>
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Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Cápsulas , Cordyceps , Ciclosporina , Medicamentos Herbarios Chinos , Usos Terapéuticos , Trasplante de Riñón , TacrolimusRESUMEN
<p><b>OBJECTIVE</b>To construct a replication-incompetent recombinant adenovirus mediating short hairpin RNA (shRNA)-induced tissue factor gene silencing in the islet.</p><p><b>METHODS</b>Four pairs of complementary oligonucleotides were designed and synthesized to create double-stranded oligonucleotides (ds oligo). The ds oligos were cloned into Pentr/U6 vector to construct the shuttle plasmid pENTR/U6-shRNA, which was transduced into human islets via liposome after sequence verification. The plasmid with the best silencing effect was identified by real-time RT-PCR, followed by homologous recombination with the adenovirus backbone plasmid. The functional clone was transfected into 293A cells to amplify the adenovirus, whose silencing effect against TF expression was tested using real-time RT-PCR and Western blotting.</p><p><b>RESULTS</b>The pENTR/U6-shRNA shuttle plasmid was constructed and verified by sequencing. The recombinant adenovirus-mediated shRNA against TF was constructed, and real-time RT-PCR and Western blotting demonstrated that the strongest silencing effect of the adenovirus against TF occurred on the 4th day following islet transfection.</p><p><b>CONCLUSION</b>Replication-incompetent recombinant adenovirus-mediated shRNA against TF has been successfully constructed, which has good silencing effect against TF expression in human islet in vitro.</p>
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Humanos , Adenoviridae , Genética , Fisiología , Secuencia de Bases , Línea Celular , ADN Recombinante , Genética , Expresión Génica , Ingeniería Genética , Métodos , Secuencias Invertidas Repetidas , Islotes Pancreáticos , Metabolismo , Plásmidos , Genética , Interferencia de ARN , ARN Interferente Pequeño , Genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tromboplastina , Genética , Carga Viral , Replicación ViralRESUMEN
<p><b>OBJECTIVE</b>To study the impacts of kidney transplantation on erectile function and analyse its contributing factors.</p><p><b>METHODS</b>In order to evaluate the severity of erectile dysfunction (ED), a total of 250 married male kidney transplant recipients (KTR) with functioning graft were assessed with the International Index of Erectile Function (IIEF) questionnaire. Data of clinical characteristics, medical and sexual history and laboratory examination were collected. Univariate and multivariate logistic regression analyses were carried out to determine which have independent impacts on erectile function.</p><p><b>RESULTS</b>The investigation was accomplished in 84.8% of the KTRs. There was no significant difference in ED incidence before and after renal transplantation (53.8% vs. 44.3%, P > 0.05). According to the IIEF score, erectile function improved in 43.9% of the KTRs, remained unchanged in 42.9%, and deteriorated in 13.2%, as compared with pre-transplantation. Logistic regression analysis showed that significant and independent influencing factors in erectile function were age, hemoglobin level, presence of DM and/or peripheral neuropathy and iterative transplantations, and their relative risks were 3.01, 2.01, 3.15, 3.89 and 2.67, respectively.</p><p><b>CONCLUSION</b>ED is highly prevalent among KTRs and its pathogenesis is multifactorial. Age, presence of DM and/or peripheral neuropathy, hemoglobin level and iterative transplantations were chief contributing factors in erectile function.</p>
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Adulto , Humanos , Masculino , Persona de Mediana Edad , Complicaciones de la Diabetes , Disfunción Eréctil , Epidemiología , Trasplante de Riñón , Modelos Logísticos , Factores de Riesgo , Encuestas y CuestionariosRESUMEN
<p><b>OBJECTIVE</b>To establish a stable method for obtaining large quantity of highly purified immature dendritic cells (imDCs) in vitro, and identify the morphology, function and surface markers of the cells.</p><p><b>METHODS</b>CD117(+) hemopoietic stem cells (HSCs) were isolated and purified from the bone marrow of healthy C57 mice by magnetic affinity cell sorting. After cell expansion by treatment with stem cell factor (SCF) and interleukin-3 (IL-3), the HSCs were induced for directional differentiation into imDCs by treatment with GM-CSF, IL-4 and IL-10. The imDCs obtained were identified by morphological and functional observation under inverted microscope, scanning electron microscope and transmission electron microscope, followed by detection of the expressions of the surface markers using flow cytometry.</p><p><b>RESULTS</b>After 3, 5 and 7 days of culture in the presence of SCF+IL-3, the cells were expanded by 10.34-/+1.43, 22.65-/+2.71 and 54.39-/+3.08 folds, respectively. The HSCs were successfully induced to differentiate into imDCs with phagocytotic activity. The dendrites of the imDCs were short small, and appearing spinous. The expressions of surface markers were detected from the cells showing the phenotype of CD11c(+), I-A/I-E(low), CD40(-), CD80(-), CD86(-).</p><p><b>CONCLUSION</b>The method described allows steadily acquisition of large quanty of highly purified imDCs and of their effective identification in vitro.</p>
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Animales , Ratones , Técnicas de Cultivo de Célula , Métodos , Diferenciación Celular , Separación Celular , Métodos , Células Cultivadas , Células Dendríticas , Biología Celular , Citometría de Flujo , Células Madre Hematopoyéticas , Biología Celular , Ratones Endogámicos C57BL , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Proteínas Proto-Oncogénicas c-kitRESUMEN
<p><b>OBJECTIVE</b>To study the protective effect of recombinant adenovirus-mediated human cytosolic glutathione peroxidase (hCGPx) gene transfection on vascular endothelial cells ECV304 from oxidative damage.</p><p><b>METHODS</b>pGEM-T Easy Vector containing hCGPx cDNA and recombinant adenovirus shuttle plasmid pACCMV-pLpA were used to construct the shuttle plasmid pACCMV-hCGPx for cotransfection of 293 cells with pJM17, thereby to obtain the recombinant adenovirus AdCMV-hCGPx. Cultured ECV304 cells were transfected with AdCMV-hCGPx for 24, 48 and 72 h, respectively, with the cells transfected with the empty vector serving as control, and hCGPx gene expression was then examined in the transfected cells. The transfected cell viability and apoptotic cell ratio were evaluated after treatment of the cells with H(2)O(2).</p><p><b>RESULTS</b>The expression ratio of hCGPx gene was significantly higher in the AdCMV-hCGPx-transfected cells than in those with empty vector transfection (P<0.01). The hCGPx gene-transfected cells showed significantly higher viability and significantly lower apoptotic ratio than the control cells following challenge with H(2)O(2)-induced oxidative damage.</p><p><b>CONCLUSION</b>hCGPx gene transfer mediated by recombinant adenovirus protects the vascular endothelial cells from oxidative damage in vitro, possibly due to the antioxidative and apoptosis-inhibiting effect of hCGPx.</p>
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Humanos , Adenoviridae , Genética , Apoptosis , Línea Celular , Supervivencia Celular , Citosol , Células Endoteliales , Biología Celular , Metabolismo , Citometría de Flujo , Vectores Genéticos , Glutatión Peroxidasa , Genética , Peróxido de Hidrógeno , Farmacología , Estrés Oxidativo , Plásmidos , Genética , Factores de Tiempo , TransfecciónRESUMEN
<p><b>OBJECTIVE</b>To investigate the role of simultaneous blockade of CD40/CD40L and B7/CD28 pathways in the immune tolerance via co-expression of sCD40LIg and CTLA4Ig mediated by replication-defective adenovirus.</p><p><b>METHODS</b>Ad-sCD40LIg-IRES(2)-CTLA4Ig, replication-defective adenovirus co-expressing sCD40LIg and CTLA4Ig, was constructed and identified. The co-expression of sCD40LIg and CTLA4Ig was evaluated with confocal laser scanning microscope and Western blotting. Skin transplantations of C57BL/6 to BALB/c mice were performed. PBS, Ad-Shuttle-CMV and Ad-sCD40LIg-IRES(2)-CTLA4Ig were administered. Skin graft survival was monitored and the mRNA expression of both genes was evaluated in the skin allografts.</p><p><b>RESULTS</b>Ad-sCD40LIg-IRES(2)-CTLA4Ig was constructed successfully and identified. The co-expression of sCD40LIg and CTLA4Ig was identified with confocal laser scanning microscopy and Western blotting. Compared to the skin graft mean survival time (MST) of non-treated group ((5.75+/-0.71) d) or Ad-Shuttle-CMV-treated group ((5.50+/-0.53) d), the skin graft MST was dramatically prolonged in the Ad-sCD40LIg-IRES(2)-CTLA4Ig-treated group ((16.38+/-1.19) d, P<0.001). The mRNA expression of both genes was detected.</p><p><b>CONCLUSION</b>Ad-sCD40LIg-IRES(2)-CTLA4Ig, a replication-defective adenovirus carrying genes encoding sCD40LIg and CTLA4Ig, was constructed. Simultaneous blockade of CD40/CD40L and B7/CD28 costimulatory pathway mediated by replication-defective adenovirus significantly prolonged skin allograft survival in mice.</p>
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Animales , Ratones , Abatacept , Adenoviridae , Genética , Efecto Citopatogénico Viral , Supervivencia de Injerto , Alergia e Inmunología , Inmunoconjugados , Genética , Alergia e Inmunología , Metabolismo , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , ARN Mensajero , Genética , Proteínas Recombinantes de Fusión , Genética , Alergia e Inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Trasplante de Piel , Alergia e Inmunología , Métodos , TransfecciónRESUMEN
<p><b>BACKGROUND</b>Globally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered 'nurse cells' in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro.</p><p><b>METHODS</b>Adult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets' function when they were co-cultured for 21 days in vitro.</p><p><b>RESULTS</b>In cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P > 0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P < 0.01).</p><p><b>CONCLUSIONS</b>A method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets' function when they were co-cultured in vitro. They could be a helper cell in islet transplantation.</p>
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Adulto , Humanos , Masculino , Separación Celular , Métodos , Supervivencia Celular , Células Cultivadas , Técnicas de Cocultivo , Islotes Pancreáticos , Fisiología , Trasplante de Islotes Pancreáticos , Células de Sertoli , Biología Celular , FisiologíaRESUMEN
<p><b>OBJECTIVE</b>To evaluate the effect of HSGJ on chronic allograft nephropathy (CAN) using standard rat model of CAN.</p><p><b>METHOD</b>Renal transplantation was performed with Fisher rats as donors and Lewis rats as recipients. All the recipients were randomly divided into control group and medication groups (high and low dosage of HSGJ, fed every other day). After 16 weeks of treatment, renal function and the histological alteration of CAN were measured. The expression of the TGFbeta1 mRNA in the allograft was evaluated by real-time PCR.</p><p><b>RESULT</b>The content of 24 h urine protein and the level of serum creatinine in the medication groups were significantly decreased (P < 0.01) as compared with control group, whereas the creatinine clearance was increased (P < 0.01). The degree of glomerular sclerosis and the Banff score of medication groups were lower than the control group respectively (P < 0.01), in consistent with decreased expression of the TGF 1mRNA.</p><p><b>CONCLUSION</b>HSGJ can prevent the chronic allograft nephropathy and the mechanism may be related with its influence on the expression of the TGFbeta1.</p>
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Animales , Ratas , Enfermedad Crónica , Medicamentos Herbarios Chinos , Usos Terapéuticos , Glomerulonefritis , Alergia e Inmunología , Rechazo de Injerto , Quimioterapia , Inmunosupresores , Usos Terapéuticos , Trasplante de Riñón , Distribución Aleatoria , Ratas Endogámicas F344 , Ratas Endogámicas Lew , Trasplante HomólogoRESUMEN
Aim To monitor the whole blood trough concentration of cyclosporine A (CsA) in renaltransplant recipients reciving triple therapy with mycophenolate mofetil, cyclosporine andprednisone and to establish an optimal therapeutic window of CsA. Methods Sampleswere measured by specific fluorescence polarization immunoassay. According to the timeafter operation and different therapy plan, the whole blood trough concentration of CsA ineach group was compared with that in control group.Results The optimal therapeuticwindow of CsA with MMF plan was 150~300 ?g? L-1 (less than one month after op-eration), 120~260?g?L-1 (1~