The Hippo pathway is generally understood to inhibit tumor growth by phosphorylating the transcriptional cofactor YAP to sequester it to the cytoplasm and reduce the formation of YAP-TEAD transcriptional complexes. Aberrant activation of YAP occurs in various cancers. However, we found a tumor-suppressive function of YAP in clear cell renal cell carcinoma (ccRCC). Using cell cultures, xenografts, and patient-derived explant models, we found that the inhibition of upstream Hippo-pathway kinases MST1 and MST2 or expression of a constitutively active YAP mutant impeded ccRCC proliferation and decreased gene expression mediated by the transcription factor NF-κB. Mechanistically, the NF-κB subunit p65 bound to the transcriptional cofactor TEAD to facilitate NF-κB-target gene expression that promoted cell proliferation. However, by competing for TEAD, YAP disrupted its interaction with NF-κB and prompted the dissociation of p65 from target gene promoters, thereby inhibiting NF-κB transcriptional programs. This cross-talk between the Hippo and NF-κB pathways in ccRCC suggests that targeting the Hippo-YAP axis in an atypical manner-that is, by activating YAP-may be a strategy for slowing tumor growth in patients.

Background: The study aimed to find predictive biomarkers to evaluate donor kidney function to predict graft dysfunction as well as to assess an early signs of acute graft rejection.Method: Twenty-seven deceased donors and 54 recipients who underwent a successful kidney transplantation were enrolled in the study. An assessment was made in serum and urine from donors and recipients to measure the following biomarkers: neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), tissue inhibitor of metalloproteinase 2 (TIMP-2) and urinary N-acetyl-b-D-glucosaminidase (uNAG). These bio-markers were used to establish a model for predicting a reduced graft function (RGF) classified as either a delayed or slow graft function.Result: Our analysis suggest that out of four tested biomarkers, the serum TIMP-2 and uNAG levels of the donors had a predictive value for RGF; the area under the receiver operating characteristic curves (AUROC) of serum TIMP-2 and uNAG were 0.714 and 0.779, respectively. The combined best fitting prediction model of serum TIMP-2, uNAG, and creatinine levels was better in predicting RGF than the serum creatinine level alone. In addition, the recipient serum TIMP-2 level on the third day post-transplantation (D3) was associated with the estimated glomerular filtration rate (eGFR) on the seventh day post-transplantation (D7; OR 1.119, 95% CI 1.016-1.233, p = 0.022). Furthermore, the ROC curve value revealed that the AUROC of TIMP-2 on D3 was 0.99 (95% CI 0.97-1, p < 0.001), and this was the best predictive value of the renal function on D7.Conclusions: Donor serum TIMP-2 and uNAG levels are useful predictive biomarkers because they can provide the donor-based prediction for RGF.

Ligustrazine is a Chinese herb (Chuanxiong) approved for use as a medical drug in China. Recent evidence suggests that ligustrazine has prom-ising antitumor properties. Our preliminary results showed that ligustrazine could inhibit the growth of human renal cell carcinoma (RCC) cell lines. However, the complicated molecular mechanism has not been fully revealed. Therefore, the purpose of this study to investigate the mech-anism of ligustrazine resistance in human RCC cells. Cell proliferation, migration, invasion, and colony-formation ability of RCC cells A498 were detected by MTT assay, clonal formation rates, and transwell chamber assay in vitro. The expression of epithelial-mesenchymal transition (EMT)-related proteins were analyzed using western blot test. The effect of ligustrazine on the growth of A498 cells in nude mice was investi-gated in vivo. Our results showed that ligustrazine could significantly inhibit the proliferation, migration, and invasion of A498 both in vivo and vitro. Western blot analysis showed that the expressions of EMT-related, N-cadherin, snail, and slug proteins were significantly decreased in A498 in the ligustrazine treatment group. This study indicated that ligustrazine could significantly inhibit the malignant biological behaviors of RCC cell lines, possibly by inhibiting the EMT process.

The shortage of transplantable organs impedes the development of tissue-engineered alternatives. Producing tissues similar to immature kidneys from pluripotent stem cells is possible in vitro, but the size of the organoids is limited. Furthermore, in vivo implantation is necessary for organoid development and functional maturation. In the present study, the induction procedure was optimized and kidney organoids derived from induced pluripotent stem cells in vitro were produced. The kidney organoids were examined by immunofluorescence and quantitative PCR. Then, a unilateral nephrectomy model was established that was beneficial to the compensatory proliferation of the other kidney. Finally, these organoids were implanted below the kidney capsules of immunodeficient mouse hosts that had been nephrectomized unilaterally. This implantation resulted in the enlargement of the organoids and the production of vascular cells. Although signs of organoid maturation were lacking in short-term culture in vivo, the present study provided a method for studying kidney organoid development in vivo.

Aryl hydrocarbon receptor nuclear translocator (ARNT) is a transcription factor that has been reported to play a vital role in regulating glycolysis, angiogenesis and apoptosis. Recently, ARNT has been reported to a play role in pancreatic-islet function in type 2 diabetes. However, the role of ARNT in kidney cancer has not yet been investigated. In the present study, ARNT expression was detected in tissues from patients with renal cell carcinoma (RCC) and in RCC cell lines. Oncomine, The Cancer Genome Atlas and cBioPortal were used to investigate the roles of ARNT in RCC. Cell migration and invasion assays were used to explore the molecular mechanisms involved. It was found that ARNT protein expression was elevated both in tissues from patients with clear cell RCC (ccRCC) and in different RCC cell lines. ARNT disruption using siRNA knockdown inhibited the migratory abilities and cell proliferation, potentially by altering the glycolysis pathway in vitro, as evidenced by decreased M2 type acetone kinase, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 and hexokinase 2 expression. Taken together, the findings in the present study revealed a novel function of ARNT in ccRCC and indicated that ARNT promotes the proliferation and invasion of ccRCC, possibly through changes to the glycolytic pathway.

Background Embryonic metanephros is the mammalian renal anlagen, which is considered as a potential source for the regeneration of functional whole kidneys. Some studies reported that metanephros implanted into unilateral nephrectomized animals can develop into kidney tissue. However, kidneys are nephrotoxic in renal failure patients, and whether metanephros can grow in nephrotoxic has not been reported. This study aims to investigate the growth of metanephros in acute nephrotoxic environment and analyze the therapeutic effect of metanephros microenvironment on acute kidney injury (AKI). Methods AKI was induced in 200 g Wistar rats by giving intramuscular injections of 50% glycerol (10 mL/kg) in their hind limbs. 45 rats were divided randomly into three groups (control, glycerin, and metanephros). Metanephros group was transplanted two metanephroi (embryonic day 15) into the renal capsule of AKI rats. Glycerin group was AKI rats without transplantation. Control group was untreated. Results Mature glomeruli and tubules were detected in the grafts in metanephros group, which means that metanephroi can grow into tissues with mature kidney structure under acute nephrotoxic. Then, we assessed the renal function of host rats and found that there were fewer tubular necrosis in metanephros group than glycerin group, and the serum creatinine and urea nitrogen were significantly lower in metanephros group than glycerin group. Conclusion These results suggested that embryonic metanephroi can grow into tissues with mature kidney structure under acute nephrotoxic, and the graft microenvironment was effective in inhibiting the progression of AKI, which provides a new approach for the treatment of acute renal injury.

Purpose: Pulmonary large-cell neuroendocrine carcinoma (LCNEC) is classified as non-small-cell lung cancer, but has characteristics similar to small-cell lung cancer. This study was performed to evaluate the effect of surgery and radiotherapy on patients with LCNEC. Materials and Methods: We analyzed 1,619 patients with stage I-III LCNEC, identified from the Surveillance, Epidemiology, and End Results database, diagnosed from 2000 to 2013. The Kaplan-Meier analysis and the Cox proportional hazard model were used to study patient prognosis. Results: Overall, 869 (53.7%) stage I LCNEC patients, 203 (12.5%) stage II patients, and 547 (33.8%) stage III patients were included in the analysis. Various surgery types were all associated with higher overall survival (OS) and lung cancer-specific survival (LCSS) than no surgery, with the following HRs: 0.334 (OS) and 0.279 (LCSS) for lobectomy, 0.468 (OS) and 0.416 (LCSS) for partial/wedge/segmental resection, and 0.593 (OS) and 0.522 (LCSS) for pneumonectomy (all p < 0.05). OS and LCSS of stage I and II LCNEC patients were not improved by radiotherapy (stage I: OS p = 0.719, LCSS p = 0.557; stage II: OS p = 0.136, LCSS p = 0.697). However, in stage III patients, radiotherapy significantly improved both OS and LCSS (p < 0.001). Following multivariate analysis, increased age, male patients, radiotherapy and diagnosed at stage II or III were all independent risk factors for LCNEC (all p < 0.05). Conclusion: Lobectomy had the best outcome for OS and LCSS in stage I-II LCNEC. For stage III LCNEC patients, radiotherapy can significantly improve survival time. However, in LCNEC patients undergoing surgery, radiotherapy may reduce survival time.

Hydroxyacyl-CoA dehydrogenase alpha subunit (HADHA) is a key lipid metabolic enzyme with a novel role in carcinogenesis. We previously reported that HADHA, a prognostic marker, was downregulated in clear cell renal cell carcinoma (ccRCC). Herein, the tumor inhibitory role of HADHA overexpression in ccRCC was investigated further. The quantitative proteomic analysis displayed that a total of 1293 and 1293 proteins were identified in HADHA overexpressed 786-O-hadha and vector-transfected control 786-O-vc cells, respectively, and 206 proteins were found to be up- or downregulated. PANTHER, OmicsNet, STRING, and DAVID tools were utilized on the dysregulated proteins in order to elucidate multiple metabolic pathways (especial lipid metabolism) and lipid metabolism-related proteins (e.g. ACAT1, ACLY). The dysregulation of the lipid metabolic enzymes, ACAT1, ACLY, CYB5R3 and FASN, were confirmed by Western blotting. Further assays demonstrated that HADHA overexpression significantly inhibited cell growth, induced cell apoptosis, and decreased the formation of cytoplasmic lipid droplets (LDs); moreover, it also inhibited tumor growth and lessened the formation of LDs in xenografted mouse. Collectively, these data revealed that HADHA overexpression disrupted lipid metabolism and inhibited tumor growth, which shed light on HADHA as a potential therapeutic target for clinical intervention of ccRCC.

OBJECTIVES:Transplant renal artery stenosis is a frequently recognized complication of kidney transplant procedures. A single-center retrospective study was conducted to examine the use of 3-dimensional computed tomography reconstruction in diagnosing transplant artery stenosis.MATERIALS AND METHODS:During 2013 at our center, 86 patients underwent kidney transplant. All patients underwent ultrasonographic analyses. Patients with clinically suspected transplant renal artery stenosis were examined by 3-dimensional computed tomography reconstruction and were treated with endovascular approaches or medically managed.RESULTS:Ten patients were diagnosed with transplant renal artery stenosis by 3-dimensional computed tomography reconstruction. No evidence of contrast-induced nephrotoxicity was observed. Nine of the 10 patients underwent percutaneous transluminal angio-plasty, whereas the remaining patient was treated conservatively. Procedural success rate was 100%. Patients were followed for a mean period of 20 ± 3 months. Blood pressure improved from a mean of 163/90 to 132/73 mm Hg at the end of the follow-up period. In the 9 patients who underwent angioplasty, serum creatinine improved from 198 ± 24 to 134 ± 16 μmol/L at the end of the follow-up period. The cystatin C level in some patients declined after interventional therapy.CONCLUSIONS:Three-dimensional computed tomography reconstruction is a safe choice for patients who present with increased serum creatinine levels and refractory hypertension. Percutaneous transluminal angioplasty is the preferred therapeutic technique for transplant renal artery stenosis.

Abstract Objective: Docetaxel (DTX) remains the only effective drug for prolonging survival and improving quality of life of metastatic castration-resistant prostate cancer (mCRPC) patients. Combination anticancer therapy encapsulating DTX and another extract of traditional Chinese medicine is one nano-sized drug delivery system promising to generate synergistic anticancer effects, to maximize the treatment effect, and to overcome multi-drug resistance. The purpose of this study is to construct lipid–polymer hybrid nanoparticles (LPNs) as nanomedicine for co-encapsulation of DTX and curcumin (CUR). Methods: DTX and CUR co-encapsulated LPNs (DTX-CUR-LPNs) were constructed. DTX-CUR-LPNs were evaluated in terms of particles size, zeta potential, drug encapsulation, and drug delivery. The cytotoxicity of the LPNs was evaluated on PC-3 human prostate carcinoma cells (PC3 cells) by MTT assays. In vivo anti-tumor effects were observed on the PC3 tumor xenografts in mice. Results: The particle size of DTX-CUR-LPNs was 169.6 nm with a positive zeta potential of 35.7 mV. DTX-CUR-LPNs showed highest cytotoxicity and synergistic effect of two drugs in tumor cells in vitro. In mice-bearing PC-3 tumor xenografts, the DTX-CUR-LPNs inhibited tumor growth to a greater extent than other contrast groups, without inducing any obvious side effects. Conclusion: According to these results, the novel nanomedicine offers great promise for the dual drugs delivery to the prostate cancer cells, showing the potential of synergistic combination therapy for prostate cancer.

Kidney transplantation is now one of the most effective treatment for end-stage renal failure. However, organ shortage remains a major problem. Hence, an alternative solution for the limited donor organs is in great need. Xenotransplantation of metanephros and cell-scaffold technology (CST) provide new ways for kidney regeneration. The structure and function of regenerated kidneys using these two methods are compared to identify the trend for kidney regeneration. Kidney scaffolds, which were decellularized by SDS perfusion, were recellularized with human umbilical vein endothelial cells (HUVEC) and mouse embryonic stem (ES) cells. Metanephros from embryonic day 13.5 were transplanted into the omentum of adult rats. After cultured in vivo for a period, the structural integrity was evaluated. The quantity and type of cells of kidney generated from metanephros were much more than that of the kidney regenerated from CST. But the latter was more suitable for transplantation. Both kidney regeneration methods have their own advantages, and one method which could offer all these advantages may be the optimal way for kidney transplantation.

The standard two dose protocol of basiliximab is effective in reducing acute rejection rate in kidney transplantation, but very expensive. This study aims to investigate the efficacy and safety of one dose of basiliximab in the prevention of acute rejection in renal transplant recipients. A total of 28 renal transplant recipients treated with one dose of basiliximab were studied and followed for six months as a non-randomized retrospective study, a simultaneous 28 renal transplant recipients without anti-lymphocyte induction therapy served as control group. All the grafts were from donation after cardiac death. Acute rejection rate was lower in the study group compared with the control group (21% vs. 50%, P < 0.05). All acute rejection episodes were reversed by methylprednisolone pulse therapy or Anti-human T Lymphocyte Porcine Immunoglobulin (ALG) therapy. There was no significant difference (P > 0.05) in six-month patient or kidney survival rate between the treatment group and the control group. Difference of adverse events including gastrointestinal dysfunctions, leukopenia, impaired liver function and infection between two groups was not found. One dose regimen of basiliximab was effective in the prevention of acute rejection in renal transplant recipients, and is fit for the situation of China because of the significant reduction of cost.

Hypospadias is a common male genital malformation and is regarded as a complex disease affected by multiple genetic as well as environmental factors. In a previous genome-wide scan for familial hypospadias, we reported suggestive linkage in nine chromosomal regions. We have extended this analysis by including new families and additional markers using non-parametric linkage. The fine mapping analysis displayed an increased LOD score on chromosome 8q24.1 and 10p15 in altogether 82 families. On chromosome 10p15, with the highest LOD score, we further studied AKR1C2, AKR1C3 and AKR1C4 involved in steroid metabolism, as well as KLF6 expressed in preputial tissue from hypospadias patients. Mutation analysis of the AKR1C3 gene showed a new mutation, c.643G>A (p.(Ala215Thr)), in a boy with penile hypospadias. This mutation is predicted to have an impact on protein function and structure and was not found in controls. Altogether, we homed in on four chromosomal regions likely to harbor genes for hypospadias. Future studies will aim for studying regulatory sequence variants in these regions.

Objective To investigate the pharmacokinetic (PK) characteristics of enteric-coated mycophenolate sodium (EC-MPS) and area under the curve (AUC) from 0 to 12 h by limited sampling strategies (LSSs) in de novo kidney transplant patients of Chinese ethnicity.Method The PK data from 20 Chinese renal transplant recipients collected during the maintenance period after transplantation were analyzed retrospectively.Ten blood samples were obtained after EC-MPS was given on the postoperative day 21.Blood concentrations of mycophenolic acid were measured and AUC was estimated using high performance liquid chromatography in recipients receiving immunosuppressant EC-MPS plus tacrolimus and steroid.LSSs with jackknife technique,multiple stepwise regression analysis,and Origin 7.5 analysis were developed to estimate mycophenolic acid (MPA) AUC.Result There was wide inter-individual variation in MPA plasma level and the AUC.The best equations obtained in the test group were for 3 and 4 time points:AUC0-12h (mg· h· L-1) =12.820 + 0.662 × C1.5h + 1.750 × C4h + 4.213 × C8h,and AUC0-2h (mg· h· L-1) =12.119 + 0.676 × C1.5h + 1.685 × C4h + 1.429 × C6h + 2.573 × C8h respectively.When these LSSs were.tested in the validation group,the results were acceptable (R =0.861,P＜0.001 for 3 time points,and R =0.882,P＜0.001 for 4 time points).Conclusion LSSs are useful for MPA AUC0-12h estimation in Chinese renal transplantation patients.LSSs using time points at C1.5h-C4h-C8h or C1.5h-C4h-C6h-C8h provide the most reliable and accurate estimation of the AUC0-12h of mycophenolic acid in Chinese stable renal transplant recipients receiving EC-MPS and tacrolimus.

Nat. Genet. 43, 48–50 (2011); published online 28 November 2010; corrected after print 24 February 2011 In the version of this article initially published, the third and fourth column headings in Table 2 were mislabeled and the abbreviations in the footnotes of Table 2 were inadvertently duplicated.The correct heading for the third column is “T” and the correct heading for the fourth column is “NT”.

Hypospadias is a common congenital malformation in boys in which the urethral meatus opens on the underside of the penis. It is considered a complex disorder with several genes involved and the molecular etiology is just beginning to be revealed. As more than 85% of Opitz G/BBB syndrome (OS) patients with MID1 mutations are manifested with hypospadias, we have investigated the association between the MID1 gene and hypospadias. DNA from 114 hypospadias cases was analyzed with direct sequencing of the MID1 gene. Genotyping analysis was performed for the single-nucleotide polymorphism (SNP) c.1230G>A in 370 individuals with varying degrees of hypospadias and compared with 759 healthy controls. We identified one nonsense mutation c.712G>T (p.E238X), one missense mutation c.1679A>G (p.K560R) and two synonymous variants c.1230G>A (p.S410S) and c.1284T>G (p.V428V). We also detected a significant difference in the rare allele frequency of SNP c.1230G>A in hypospadias patients as compared with controls (P=0.016). Our finding suggests that hypospadias associated with hypertelorism is the mildest phenotype in OS caused by MID1 mutations.

Objective To study the expressions of Th17 lymphocytes related cytokines including interleukin 17A(IL17A) and interleukin 23(p19/p40)(IL23p19/p40) in prostate tissue of experimental autoimmune prostatitis(EAP) of C57BL/6 mice. Methods Prostate glands from Wistar rats were used to make purified prostate protein,and then the mice were subcutaneously injected with the purified protein combined with complete Freund′s adjuvant(CFA) and diphtheria pertussis tetanus (DPT) vaccine at 0 and 30 days.30 days after the second injection the prostates of the immune mice were observed,and one-step real-time RT-PCR was used to check mRNA expressions of Th17 lymphocytes related cytokines including IL17A and IL23(p19/p40) in the prostate tissue of EAP mice,and the differences of the cytokines between the EAP and control groups were compared to study the effectiveness of the cytokines in the onset of autoimmune prostatitis of C57BL/6 mice.Results The mice model of EAP was successfully made.The mRNA relative amounts of IL17A,IL23p19 and IL23p40 in the mice prostate of the EAP and the control group were 0.053 0±0.026 5 vs 0.017 7±0.006 5,0.028 9±0.028 3 vs 0.003 6±0.002 9,0.005 1±0.002 5 vs 0.004 1±0.002 8 respectively,and the differences between the two groups were significant(all P0.05).Conclusion Expressions of Th17lymphocytes related cytokines in the prostate tissue of EAP mice are higher than those in the controls,and these cytokines are related to the onset of EAP.

OBJECTIVES:The sympathetic nervous system and hypothalamic-pituitary-adrenal axis have been known to be reactive to psychological factors such as stress in which several humoral factors are released. Growing data suggest that stress conditions could be associated with the development or aggravation of prostatic disease. The present study aims to investigate the influence of chronic stress and blockade of the sympathetic nerve on rat prostate.METHODS:We used 2 weeks of restraint water-immersion stress to induce a chronic psychological stress model of Wistar rats. Denervation of the peripheral sympathetic nerve was done by 6-hydroxydopamine (6-OHDA). Blood flow of ventral prostate, plasma catecholamine and cortisol were measured at the end of the experiment. The weight and secretions of dissected prostate lobes were measured and histology was performed by hematoxylin and eosin stain.RESULTS:After 14 days of stress, the blood flow of ventral prostate decreased and plasma cortisol increased. The apparent proliferation of epithelium of ventral lobes were observed and dorsolateral ones were almost unaffected. On the other hand, the hyperplasia changes were not seen after the chemical sympathectomy had been done during stress.CONCLUSIONS:Chronic stress may lead to hyperplasia of ventral prostate, the overactivity of sympathetic nervous system is involved in the pathogenesis of prostatic disease.

To analyze etiology, clinical features, effective diagnostic methods and therapeutic efficacy of infant urolithiasis related to melamine.