Stromal cell-derived factor 1 gene polymorphism is associated with susceptibility to adverse long-term allograft outcomes in non-diabetic kidney transplant recipients.

Although the genetic polymorphism of Stromal Cell-Derived Factor 1 (SDF-1) is associated with higher mortality of liver allograft recipients, the role of SDF-1 in the modulation of renal allograft outcomes is unclear. Between March 2000 and January 2008, we recruited 252 non-diabetic renal transplant recipients (RTRs). Baseline characteristics and blood chemistry were recorded. Genomic DNA extraction with polymerase chain reaction-restriction fragment length polymorphism was utilized to analyze the genetic polymorphisms of SDF-1 (rs1801157). The influence of SDF-1 on an adverse renal allograft outcome, defined as either a doubling of serum creatinine, graft failure, or patient death was evaluated. Sixteen patients with the SDF-1 AA/AG genotype and nine with the SDF-1 GG genotype reached an adverse outcome. According to Kaplan-Meier analysis, patients carrying the SDF-1 AA/AG genotype or A allele showed a significantly higher risk of reaching an adverse outcome than those carrying the SDF-1 GG genotype or G allele (p=0.041; p=0.0051, respectively; log rank test). Stepwise multivariate Cox proportional regression analysis revealed that patients carrying the SDF-1 AA/AG genotype and A allele had a 2.742-fold (95% CI. 1.106-6.799, p=0.03) and 2.306-fold (95% CI. 1.254-4.24, p=0.008) risk of experiencing an adverse outcome. The SDF-1 AA/AG genotype and A allele have a detrimental impact on the long-term outcome of RTRs.

Subclinical peripheral arterial disease in renal transplantation.

BACKGROUND: Peripheral arterial disease (PAD) is a common finding in patients at various stages of chronic kidney disease; however, there has only been a limited amount of data that have been published regarding the prevalence and associated risk factors of subclinical PAD in renal transplant recipients. METHODS: The authors cross sectionally investigated the prevalence of PAD using ankle-brachial index (ABI) in 304 renal transplant recipients with no previous diagnosis of PAD. Patients were considered to have subclinical PAD when ABI <0.9. The authors also determined the associated risk factors for subclinical PAD. RESULTS: The mean age of the 304 patients was 53 years, and 30 patients (9.9%) had a history of atherosclerotic event (including past cardiovascular and cerebrovascular events). Twenty-five of the 304 patients (8%) had ABI <0.9 and 1 had (0.3%) ABI >1.3. Compared to patients with normal ABI, a history of atherosclerotic events is the only independent risk factor for patients with subclinical PAD (P = 0.0468). CONCLUSIONS: Subclinical PAD is an inadvertent issue in renal transplant patients, especially those with a history of atherosclerotic events. Further research is needed on the long-term clinical impact and optimal treatment of subclinical PAD among renal transplant patients.

Conversion to prolonged release tacrolimus formulation in stable kidney transplant recipients.

PRINCIPLES: The once-daily tacrolimus formulation (Advagraf®), with the potential for improving medical adherence, has been advocated to improve long-term kidney allograft outcomes. However, experience with late conversion from the twice-daily tacrolimus formulation (Prograf®) to Advagraf in the daily care of stable kidney transplant recipients has been limited. METHODS: The aim of this study was to observe the efficacy and safety of conversion from Prograf to Advagraf in chronic stable kidney transplant recipients in routine clinical practice. The recruited patients had postconversion follow-up at least once monthly for a total of six months, unless they had discontinued the use of Advagraf, had been lost the follow-up or had lost the graft. RESULTS: The mean age of the 199 patients was 51.5 ± 10.4 years (60.8% male). The mean time from transplantation to the conversion to Advagraf was 8.3 ± 3.2 years and the mean tacrolimus trough level at conversion was 4.2 ± 1.4 ng/ml. After conversion, 147 patients (73.8 %) had a reduced trough level at one month and the mean change in trough level postconversion was -13.5%. The mean serum creatinine level between conversion and six months postconversion was not significantly different (1.12 ± 0.36 vs 1.10 ± 0.42 mg/dl). Thirty-four patients (17%) discontinued the treatment with Advagraf and two (1%) developed biopsy-proved acute rejection. CONCLUSIONS: In conclusion, frequent conversion caused by a high discontinuation rate may further raise the potential risk of allograft rejection and increase unnecessary cost. In view of this, the policy of converting to Advagraf with the purpose of improving medical adherence should be individualised in routine clinical practice.

Community-acquired urinary tract infection in kidney transplantation: risk factors for bacteremia and recurrent infection.

BACKGROUND/PURPOSE: Urinary tract infection (UTI) is the most common type of infectious complication among kidney transplant patients. However, the antibiotic susceptibility of causative microorganisms and risk factors for concomitant bacteremia and recurrent infection are rarely discussed. METHODS: This was a retrospective cohort review of kidney transplant recipients who had received follow-up in the past 10 years at the Chung-Shan Medical University (Taichung, Taiwan). Only community-acquired and symptomatic UTIs were included in this study. RESULTS: During the 53 ± 22 months of follow-up, 99 patients developed 167 episodes of UTI. Forty-two (25%) episodes had concomitant bacteremia. Escherichia coli was the most common causative microorganism, and strains with resistance to multiple commonly used empirical antibiotics began to emerge. The independent risk factors for UTI with concomitant bacteremia in multivariate analysis were immunosuppression with tacrolimus (adjusted odds ratio [AOR] 3.17; 95% confidence interval [CI] 1.29-7.75; P = 0.011) and baseline serum creatinine level >1.3 mg/dL before first UTI (AOR 2.55; 95% CI 1.02-6.36; P = 0.045). However, there were no factors that were significantly associated with recurrent infection. CONCLUSION: From this study, we found that E coli tends to have resistance to commonly used empirical antibiotics in this modern era and that patients who use the immunosuppressant tacrolimus and have baseline serum creatinine level >1.3 mg/dL before their first UTI have a tendency to suffer from concomitant bacteremia and even sepsis.

A higher glomerular filtration rate predicts low risk of developing chronic kidney disease in living kidney donors.

BACKGROUND: The risk of developing chronic kidney disease (CKD) among living kidney donors (LKDs) is seldom included in evaluations of patients' outcomes. Potential risk factors and new criteria for estimating the glomerular filtration rate (eGFR) indexed for body surface area (BSA) were investigated with a view to prevent the development of CKD in LKDs. METHODS: We conducted a retrospective study of LKDs from May 1983 to March 2011. The Mann-Whitney U test and χ(2) test were used to analyze the male versus female groups. Survival analysis was plotted as CKD-free survival and analyzed separately by different eGFR index classifications. The Cox regression model was used to identify potential risk factors for development of CKD. RESULTS: A total of 105 LKDs with a mean age of 46.3 ± 12.5 years had a mean eGFR indexed for BSA of 88.9 ± 21.5 ml/min per 1.73 m(2). After a mean duration of 5.4 ± 4.9 years' follow-up, eGFR dropped to 61.4 ± 16.4 ml/min per 1.73 m(2) (p = 0.002). Median CKD-free survival was only 5.7 years. The difference between eGFR ≥ 80 ml/min per 1.73 m(2) and <80 ml/min per 1.73 m(2) was not statistically significant (p = 0.980). Multivariate Cox regression analysis showed that higher eGFR at donation (HR = 0.952, p = 0.0199) could be a protective factor. The receiver operating characteristic (ROC) curve for initial eGFR with best sensitivity of 52.78 % and specificity of 81.40 % was obtained with a cutoff value of 90.2 ml/min per 1.73 m(2) for preoperative eGFR. An eGFR of 90 ml/min per 1.73 m(2) yielded a significant survival curve (p = 0.0199) after 21 years of follow-up. Further classifications of eGFR >90 ml/min per 1.73 m(2) into 90-99 ml/min per 1.73 m(2), 100-109 ml/min per 1.73 m(2), and ≥110 ml/min per 1.73 m(2) were examined, but this survival curve was not statistically significant (p = 0.1247). CONCLUSIONS: Living kidney donors will develop CKD after a long duration of follow-up if there is insufficiently high eGFR at donation. An eGFR above 90 ml/min per 1.73 m(2) before donation is the only factor that predicts prevention of CKD. Larger studies with longer duration of follow-up are necessary to clarify the clinical outcome of this postoperative CKD group, especially for patients with eGFR between 80 and 90 ml/min per 1.73 m(2).

Glutathione S-transferase M1 gene polymorphism is associated with susceptibility to impaired long-term allograft outcomes in renal transplant recipients.

BACKGROUND: Despite improved post-transplantation care, progress in long-term kidney allograft survival of diabetic renal transplant recipients (pre-DM RTR) is worse than that of non-diabetic recipients (non-DM). We hypothesized that there are other potential risk factors, that predispose RTR to adverse renal allograft outcomes. METHODS: A total of 323 transplant recipients who underwent renal transplantation between March 2000 and January 2008 were recruited. The composite end-point consisted of serum creatinine (SCr) doubling, graft failure, and death. Baseline clinical data were recorded, and polymerase chain reaction-restriction fragment length polymorphism measurements of interleukin (IL)-4, IL-10, IL-23, glutathione S-transferase (GST)A1, GSTM1, and GSTP1 polymorphisms were determined. The risk factors for developing the primary outcome were analyzed among these clinical and genetic factors. RESULTS: Within a mean follow-up of 71.1 ± 24 months, there were 43 (13.3 %) patients with the primary outcome. Stepwise multivariate Cox regression analysis was used to determine the risk factors for the primary outcome of RTR. Renal transplant recipients who possessed the GSTM1 null genotype had a 2.2-fold risk (95 % CI: 1.10-4.40; P = 0.026) of developing the primary outcome. Additionally, RTR that had DM before transplantation (aHR: 3.31; 95 % CI: 1.77-6.20; P = 0.0002) or changes in SCr 6 to 12 months after transplantation (aHR: 2.83; 95 % CI: 1.29-6.19; P = 0.0095) had an increased risk of developing the primary outcome. CONCLUSIONS: In addition to the adverse role played by DM, the GSTM1 null genotype also has an unfavorable influence on the long-term allograft outcome of RTR.

Increased expression of intranuclear matrix metalloproteinase 9 in atrophic renal tubules is associated with renal fibrosis.

BACKGROUND: Reduced turnover of extracellular matrix has a role in renal fibrosis. Matrix metalloproteinases (MMPs) is associated with many glomerular diseases, but the histological association of MMPs and human renal fibrosis is unclear. METHODS: This is a retrospective study. Institutional Review Board approval was obtained for the review of patients' medical records, data analysis and pathological specimens staining with waiver of informed consents. Specimens of forty-six patients were examined by immunohistochemical stain of MMP-9 in nephrectomized kidneys, and the association of renal expression of MMP-9 and renal fibrosis was determined. MMP-9 expression in individual renal components and fibrosis was graded as high or low based on MMP-9 staining and fibrotic scores. RESULTS: Patients with high interstitial fibrosis scores (IFS) and glomerular fibrosis scores (GFS) had significantly higher serum creatinine, lower estimated glomerular filtration rate (eGFR), and were more likely to have chronic kidney disease (CKD) and urothelial cell carcinoma. Univariate analysis showed that IFS and GFS were negatively associated with normal and atrophic tubular cytoplasmic MMP-9 expression and IFS was positively correlated with atrophic tubular nuclear MMP-9 expression. Multivariate stepwise regression indicated that MMP-9 expression in atrophic tubular nuclei (r = 0.4, p = 0.002) was an independent predictor of IFS, and that MMP-9 expression in normal tubular cytoplasm (r = -0.465, p<0.001) was an independent predictor of GFS. CONCLUSIONS: Interstitial fibrosis correlated with MMP-9 expression in the atrophic tubular nuclei. Our results indicate that renal fibrosis is associated with a decline of MMP-9 expression in the cytoplasm of normal tubular cells and increased expression of MMP-9 in the nuclei of tubular atrophic renal tubules.

Long-term outcomes of patients treated with primary stenting for transplant renal artery stenosis: a 10-year case cohort study.

BACKGROUND: The purpose of the present study was to evaluate graft and patient survival and long-term outcomes of primary endoluminal stenting (PES) as an initial treatment for transplant renal artery stenosis (TRAS). METHODS: From December 1999 to March 2010, 744 consecutive patients undergoing renal transplantation were enrolled. Patients were divided into one of two groups: the study group, comprised of 18 patients who underwent PES for TRAS > 60%, and a control group, including the remaining 726 recipients who did not develop TRAS post-transplantation. Primary outcome measures were death-censored graft failure and all-cause mortality. The immediate and long-term effects of PES were evaluated by assessing blood pressure (BP) control and biochemical graft function. RESULTS: The technical success rate for PES was 100%, and minor complication occurred in only one case (5.6% of the study group). With a mean follow-up of 7.1 ± 3.7 and 6.9 ± 2.4 years in the study and control groups, respectively, 4 patients in the study group and 113 patients in the control group reached the primary outcome (log rank P = 0.418). The reduction in stenosis resulted in immediate improvement in BP control and graft function, which persisted throughout the 6 year follow-up period. Restenosis occurred in only one patient (5.6%), but restenosis was not the cause of graft failure. CONCLUSIONS: This study indicated that both the long-term graft and patient survival were as good in TRAS patients treated with PES as in patients without TRAS. The data also supported the use of PES as an initial treatment for TRAS.

Y27632 attenuates the aristolochic acid-promoted invasion and migration of human urothelial cancer TSGH cells in vitro and inhibits the growth of xenografts in vivo.

BACKGROUND: Aristolochic acid I (AAI) has been implicated in urothelial cell carcinoma (UCC) in humans. However, whether AAI promotes invasion/migration of UCC has not been established. METHODS: A study of human UCC TSGH cells cultured with AAI was conducted. Cell viability, the effects of AAI on the activity of matrix metalloproteinase (MMP)-9, the abilities of invasion/migration and the migration-related proteins (Ras, RhoA, ROCK1, PI-3K, pAkt and nuclear factor-kappaB) of the TSGH cells were assessed. The TSGH cells were subcategorized to 1-day or 30-day AAI exposure. An in vivo study using a nude mice xenograft model was employed to test the antitumor effects of Rho kinase inhibitor or Y27632. RESULTS: A time- and dose-dependent increase in both activity and messenger RNA (mRNA) level of MMP-9 were demonstrated. The mRNA level of urokinase-type plasminogen activator was increased and tissue inhibitor of metalloproteinase-1 was decreased in the cells with 30-day but not 1-day AAI exposure. A dose-dependent enhancement in wound-healing rate and cell migration was demonstrated, especially in the 30-day AAI-exposed cells. Expressions of Ras/RhoA and other migration-related proteins were increased after AAI treatment, which could be inhibited by Y27632. The in vivo results demonstrated that Y27632 was able to attenuate the speed of growth of the inoculated tumors in nude mice. CONCLUSION: Clinically, the patients with prolonged AAI exposure are highly associated UCC, our results provided in vitro and in vivo evidence that prolonged AAI exposure enhances invasion and migration of human TSGH cells.

Glutathione S-transferase gene polymorphisms are not major risks for susceptibility to posttransplantation diabetes mellitus in Taiwan renal transplant recipients.

Glutathione S-transferase (GST) M1 null genotype has been reported playing a significant role in the diabetes mellitus (DM) susceptibility in Turkish population. We investigated whether the GSTM1, GSTA1, and GSTP1 gene polymorphisms are associated with posttransplantation diabetes mellitus (PTDM) in Taiwan. There were 283 renal transplant recipients (RTRs) enrolled. Polymerase chain reaction-restriction fragment length polymorphism was used for the measurement of GSTA1, M1, and P1 genetic polymorphisms. PTDM was diagnosed according to the American Diabetes Association guidelines. Eight-five patients (30%) were diagnosed with PTDM. The averaged posttransplant follow-up period was 77.9 ± 27.2 months. Duration from transplantat to diagnosis of PTDM ranged from 0.2 to 103.1 months (19.2 ± 26.3 months). There were significantly differences between non-DM and PTDM groups in age (50.6 ± 11.0 vs. 54.6 ± 9.36 years, P = 0.005), BMI (22.4 ± 3.6 vs. 24.3 ± 3.8, P<0.001). The distributions of GSTA1, GSTP1, and GSTM1 genotypes alleles were not significantly different between PTDM and non-DM group. Patients carrying the different GSTA1, GSTP1, and GSTM1 genetic and allelic polymorphisms had no differences for the development of PTDM. These overall results suggested a lack of strong association with GSTA1, GSTP1, and GSTM1 genetic polymorphisms to the susceptibility of PTDM in Taiwanese RTRs.

Long-term impact of pretransplant and posttransplant diabetes mellitus on kidney transplant outcomes.

BACKGROUND: The aim of this study was to compare the impact of preexisting diabetes mellitus (pre-DM), posttransplant DM (PTDM), and non-DM on the long-term outcomes of renal transplant recipients (RTRs). METHODS: This is a retrospective observational cohort study of 427 RTRs who underwent transplantation from 1999 to 2008. Patients were divided into non-DM, pre-DM, and PTDM groups. The primary outcome was the composite of doubling of the serum creatinine (SCr) level, graft failure, or death. Secondary outcomes were biopsy-proven acute rejection (BPAR), biopsy-proven interstitial fibrosis and/or tubular atrophy (IF/TA), and individual components of the primary outcome. RESULTS: A total of 70 patients (16.4%) had pre-DM, 104 (24.2%) had PTDM, and 253 (59.3%) had non-DM. Kaplan­Meier analysis indicated significant differences in the development of the primary outcome: p = 0.003 (log rank test). Relative to the non-DM group, the pre-DM group had a 6.36-fold increased risk [95% confidence interval (CI) 2.43­16.33; p < 0.001), and the PTDM group had a 2.00-fold increased risk (95% CI 1.08­3.73; p = 0.029) for development of the primary outcome. Patients in the pre-DM group had 6.73-fold (95% CI 2.46­18.42; p < 0.001), 4.56-fold (95% CI 1.77­11.78; p = 0.002), and 13.95-fold (95% CI 2.96­65.75; p < 0.001) increased risks for the development of SCr doubling, biopsy-proven IF/TA, and death-censored graft failure, respectively. Patients in the PTDM group had a 2.09-fold (95% CI 1.10­3.99; p = 0.025), increased risk for the development of SCr doubling. CONCLUSIONS: The presence of pre-DM or PTDM significantly impaired kidney allograft outcome.

Silibinin inhibits the invasion and migration of renal carcinoma 786-O cells in vitro, inhibits the growth of xenografts in vivo and enhances chemosensitivity to 5-fluorouracil and paclitaxel.

Silibinin is a flavonoid antioxidant that is widely used for its anti-hepatotoxic properties. It exerts a dose-dependent inhibition on the invasion and migration of 786-O renal cell carcinoma (RCC) cells in the absence of cytotoxicity. 786-O cells were treated with silibinin at various concentrations, up to 50 µM, for a defined period and then subjected to gelatin zymography, casein zymography, and Western blot to investigate the impacts of silibinin on metalloproteinase (MMP) -2, -9, urokinase plasminogen activator (u-PA), and MAPK pathway signaling proteins, respectively. The results showed that silibinin decreased MMP-2, MMP-9, u-PA, p-p38, and p-Erk1/2 expressions in a concentration-dependent manner. The reduced expressions of MMP-2 and u-PA, as well as inhibition of cell invasion were obtained in the cultures pre-treated with PD98059 (Erk1/2 inhibitor) and SB203580 (p38 inhibitor). An in vivo anti-tumor study with a nude mice xenograft model by a subcutaneous inoculation of 786-O cells demonstrated small solid tumors after eight days following cell inoculation. There was a 70.1% reduction in tumor volume and 69.7% reduction in tumor weight by silibinin feeding on day 44, compared to those of controls. Moreover, combination treatment with silibinin and 5-fluorouracil, paclitaxel, vinblastine, or RAD-001 enhanced the chemosensitivity of 5-fluorouracil and paclitaxel. In conclusion, silibinin inhibits the invasion and migration of 786-O cells in vitro, inhibits the growth of xenografts in vivo, and enhances chemosensitivity to 5-fluorouracil and paclitaxel. © 2011 Wiley-Liss, Inc.

Primary ovary transitional cell carcinoma after renal transplantation.

Transitional cell carcinoma (TCC) of the urinary tract is the most frequent malignancy following renal transplantation reported in Taiwan. A 67-year-old female underwent bilateral nephrouretectomy and bladder cuff excision because of bilateral hydronephrosis 5 years after cadaveric renal transplantation. The pathologic report was only atrophied kidney. Pelvic sonography and abdominal computed tomography showed a pelvic mass 8 years after transplantation. After gynecological surgery, the pathologic report of the left ovarian tumor was TCC, high grade, stage IIA. The patient then underwent four cycles of postoperative chemotherapy with carboplatin and gemcitabine. TCC of the ovary is a rare, recently recognized subtype of ovarian surface epithelial cancer. We present the first case of primary ovarian TCC following renal transplant.

Association between interleukin 23 receptor polymorphism and kidney transplant outcomes: a 10-year Taiwan cohort study.

OBJECTIVE: Interleukin 23 receptor (IL-23R) plays a role in the pathogenesis of multiple autoimmune processes. The relationship between allograft outcomes and the IL-23Rvariant genotypes has not been reported on previously. Therefore, we examined the relationship between this genetic polymorphism and kidney transplant outcomes. METHODS: This is an observational cohort study and 422 renal transplant recipients (RTRs) were enrolled. Polymerase chain reaction-restriction fragment length polymorphism was used for the measurement of IL-23R genetic polymorphisms. We used a composite end-point incorporating serum creatinine (SCr) doubling, graft failure and death as the primary outcome. Secondary outcomes included biopsy-proven acute rejection (BPAR), biopsy-proven interstitial fibrosis/tubular atrophy (IF/TA) and individual primary outcome. The risks of developing primary and secondary outcomes were compared between the different IL-23R genotypes and alleles. RESULTS: With a mean follow-up of 79.3±28.8 months, 26 patients in the IL-23R genotype AA group and 32 patients in the IL-23R genotype AC/CC group reached the primary outcome (p=0.061). RTRs who carried the IL-23R AC/CC genotype (aHR 1.78; 95% CI. 1.01-3.12; p=0.046) and C allele (aHR 1.48; 95% CI. 0.96-2.28; p=0.075) had a higher risk of developing primary outcome as compared to those with IL-23R AA genotype and A allele, respectively. Moreover, RTRs who carried the IL-23R AC/CC genotype and C allele had a higher risk of developing biopsy-proven IF/TA (p=0.012; p=0.012) and SCr doubling (p=0.024; p=0.042) as compared to those with IL-23R AA genotype and A allele, respectively. The risk of BPAR, graft failure and death between the IL-23R genotypes and alleles were comparable. CONCLUSION: IL-23R polymorphism may have a potential immuno-modulating role in long-term allograft outcome.

Plasminogen activator inhibitor-1 5G/5G genotype is a protecting factor preventing posttransplant diabetes mellitus.

OBJECTIVE: Plasminogen activator inhibitor 1 (PAI-1) is thought to play a role in the pathogenesis of obesity and insulin resistance. A connection between gestational diabetes mellitus and the functional -675 PAI-1 genotype has been reported. Therefore, we examined the role of the PAI-1 gene polymorphism in kidney transplant recipients. METHODS: A total of 376 kidney transplant recipients were prospectively screened for posttransplant diabetes mellitus (PTDM). Eighty-one (21.5%) patients were diagnosed with PTDM and the other 295 patients were non-diabetic following kidney transplantation. DNA samples were isolated from the sera and analyzed for the functional -675 4G/5G promoter polymorphisms of the PAI-1 gene. RESULTS: Kidney transplant recipients with PTDM were significantly associated with tacrolimus use (p=0.03), older age (p=0.036), and higher body mass index (p=0.001). The genotype distribution was significantly different between the patients with PTDM (genotype 4G/4G:4G/5G:5G/5G=33.3%:60.5%:6.2%) and those without PTDM (genotype 4G/4G:4G/5G:5G/5G=36.9%:44.1%:19.0%) (p=0.018). Patients with homozygosity for 5G had a significantly lower rate of PTDM (aOR, 0.286, p=0.022) and higher cumulative event-free probability of time to PTDM (log rank test, p=0.0058). CONCLUSION: Homozygosity for the 5G allele of the PAI-1 gene constitutes a protecting factor for the development of PTDM. Our findings are similar to a previous study on gestational diabetes mellitus, and strongly support a possible genetic role of PAI-1 in the development of PTDM.

The suppressive effect of Rho kinase inhibitor, Y-27632, on oncogenic Ras/RhoA induced invasion/migration of human bladder cancer TSGH cells.

Urothelial cell carcinoma is the most common type of malignancy found in long-term dialysis patients and kidney transplant recipients in Taiwan. Surgical specimens of tumorous and non-tumorous bladder tissues were collected from 12 patients with bladder cancer. Increased expressions of Ras, RhoA, Akt, PI-3K were demonstrated in the tumors as compared to adjacent control tissues. To understand the impact of Ras over-expression on bladder cancer progression, human bladder cancer TSGH 8301 cells were transfected with Ras DNA. The Ras-transfected cells were then treated with either a PI-3K inhibitor (wortmannin) or Rho kinase inhibitor (Y-27632) and the expressions of Ras, PI-3K, Akt, NF-kappaB, and RhoA were analyzed. Fluorescent phalloidin staining demonstrated more intense F-actin staining in the Ras over-expressed cells than in the control cells, and the intensity of F-actin was inhibited by Y-27632. A gelatin zymography study demonstrated that the MMP-2 and MMP-9 expressions of the Ras-transfected cells were enhanced, and Y-27632 treatment reduced the levels of MMP-2 and MMP-9. Similarly, a wound healing assay revealed that the ability of cell migration was markedly increased by Ras transfection and the healing rate after treatment of Y-27632 was delayed. Our results provide evidence that Ras-induced RhoA and NF-kappaB activation was involved in the invasion/migration of bladder cancer. Through Ras and/or RhoA inhibition, there might be an opportunity for new therapeutic interventions in bladder cancer.

Prediction of chronic allograft damage index of renal allografts using serum level of plasminogen activator inhibitor-1.

BACKGROUND: Plasminogen activator inhibitor-1 (PAI-1) plays an important role in renal fibrosis. We conducted this study to examine whether serum PAI-1 has a role in predicting chronic allograft nephropathy (CAN). METHODS: Fifty kidney transplant recipients receiving graft biopsies were enrolled. The pathologic diagnoses were acute tubular necrosis (ATN; n = 12), borderline rejection (BR; n = 7), acute rejection (ACR; n = 12), CAN (n = 11), polyomavirus nephropathy (PVN; n = 3) and others (n = 5; glomerulopathy, n = 4; calcineurin inhibitor nephropathy, n = 1). The serum level of PAI-1 and chronic allograft damage index (CADI) score of each patient were determined. RESULTS: The CADI score of all patients was 4 (0-10) (median and range) and in each group was ATN 0.5 (0-4), BR 2.0 (1-4), ACR 4.0 (2-8), CAN 5.0 (4-10), PVN 7 (4-8) and others 2 (0-8). The serum PAI-1 level of each group was ATN 9.38 (2.35-14.52) ng/mL, BR 10.09 (0.61-18.06) ng/mL, ACR 11.08 (2.32-20.68) ng/mL, CAN 14.51 (3.66-21.12), PVN 14.79 (13.94-21.94) and others 16.35 (4.05-21.01) ng/mL. CADI score is associated with serum PAI-1 activity (r = 0.405, p = 0.003) and is inversely associated with serum creatinine (r = -0.348, p = 0.011), but not with estimated glomerular filtration rate (p = 0.124). CONCLUSION: Serum PAI-1 level has the potential to be a marker to predict CADI score, the quantitative score of CAN.

The effect of low-dose cidofovir on the long-term outcome of polyomavirus-associated nephropathy in renal transplant recipients.

BACKGROUND: Polyomavirus-associated nephropathy (PVAN) has an unfavourable impact on graft survival. The cornerstone of therapy is early reduction of immunosuppressive medications; however, the rate of graft failure is still high. Antiviral drugs, such as cidofovir, are thought to have therapeutic effects, but the benefits of cidofovir in retarding the deterioration of PVAN are still a controversial issue. METHODS: Fourteen renal kidney recipients were diagnosed to have biopsy-proven PVAN between 2001 and 2006 in Chung-Shan Medical University Center with nearly 600 renal transplant recipients. After the diagnosis of PVAN, all patients were treated with a reduction of their original immunosuppressive medications with/without converting tacrolimus to cyclosporine. Eight of the 14 patients agreed to receive low-dose cidofovir (0.5 mg/kg) every 2 weeks for a total of six doses. RESULTS: During 30 +/- 18 months of follow-up, three (37%) patients in the cidofovir-treated and three (50%) patients in the non-cidofovir-treated group experienced graft loss (P = 0.64). The rejection rate before PVAN diagnosis or other baseline characteristics of the patients between two groups were not significantly different. The long-term survival rate to graft loss and major graft functional decline with Kaplan-Meier analysis between the two groups were not significantly different (P = 0.898 and P = 0.243). In all demographic and clinical characteristics, we found that there was a tendency towards long-term major graft functional decline in the patients with acute rejection prior to PVAN diagnosis (P = 0.04). CONCLUSIONS: We concluded that (1) there was no obvious effect of low-dose cidofovir on long-term graft survival in patients with PVAN, and (2) acute rejection prior to PVAN diagnosis was a potential risk factor for poorer long-term graft outcome.

Impact of plasminogen activator inhibitor-1 gene polymorphisms on primary membranous nephropathy.

BACKGROUND: Idiopathic membranous nephropathy (MN) is one of the most common causes of nephrotic syndrome in adults, and 25% of MN patients proceed to end-stage renal disease. Plasminogen activator inhibitor type 1 (PAI-1) activity plays an important role in renal fibrosis. The objective of this study was to clarify the relationship between PAI-1 gene polymorphisms and the progression of MN-associated pathologies. METHODS: We recruited a cohort of 104 biopsy-diagnosed MN patients and 142 healthy subjects that served as controls. Genotyping of PAI-1 gene polymorphisms was performed using allele-specific polymerase chain reaction methods. We then analysed associations between PAI-1 gene 4G/5G polymorphisms and clinical manifestations and progression of MN. RESULTS: The genotype distribution had no effect on the development of MN. The last measured creatinine clearance in MN patients having the 4G/4G genotype was significantly lower than in patients having the 4G/5G or 5G/5G genotypes (43.6 +/- 33.6, 55.8 +/- 44.3 and 73.3 +/- 29.8 ml/min, respectively, P = 0.008). Coronary artery diseases were more prevalent in patients having the 4G5G (14/32%) and 4G4G genotypes (4/11%) than in those having the 5G5G genotype (1/5%, P = 0.008). Peripheral vascular events were more prevalent in patients having the 4G5G (18/41%) and 4G4G (6/16%) genotypes than in those having the 5G5G genotype (3/14%, P = 0.021). Disease progression occurred more frequently in patients having the 4G4G (20/53%) and 4G5G (25/57%) genotypes compared with those having the 5G5G genotype (5/23%, P = 0.026). CONCLUSIONS: The presence of the 4G allele was associated with renal deterioration and increased cardiovascular as well as other vascular events in MN patients. These findings should prompt specific considerations for the treatment of MN in patients having the 4G4G genotype.

Circulating matrix metalloproteinase-2 is associated with cystatin C level, posttransplant duration, and diabetes mellitus in kidney transplant recipients.

Studies have indicated that matrix metalloproteinase-2 (MMP-2) is vital for the patient's condition after renal transplantation. Although the allograft survival rate has been improved, the relationships between various clinical parameters in stable graft function and serum MMP-2 still need to be clarified. In this study, gelatin zymography and enzyme-linked immunosorbent assay were employed to measure MMP-2 level in the plasma of 152 kidney transplant recipients, 41 chronic kidney disease patients, and 50 healthy control subjects. The creatinine and the MMP-2 levels in the transplant recipients were significantly greater (P < 0.001) than those of control subjects. Univariate and stepwise regression analysis demonstrated the MMP-2 level was associated with cystatin C level (P < 0.001), creatinine level (P = 0.036), proteinuria (P = 0.043), posttransplant days (P = 0.025), and posttransplant diabetes mellitus (P = 0.03). We conclude that circulating MMP-2 is associated with cystatin C, posttransplant duration, and diabetes mellitus in kidney transplant recipients and suggest that MMP-2 may be critical for graft survival.