VHL-TGFBI signaling is involved in the synergy between 5-aza-2'-deoxycytidine and paclitaxel against human renal cell carcinoma.

PURPOSE: To analyse the role of von Hippel-Lindau (VHL) and transforming growth factor ß-induced (TGFBI) in synergistic mechanisms of 5-aza-2'-deoxycytidine (DAC) and paclitaxel (PTX) against renal cell carcinoma (RCC). METHODS: To elucidate the role in the synergy between DAC and PTX against RCC cells, TGFBI expression was regulated using siRNA technology and an expression vector containing the full-length cDNA for TGFBI was also transfected into RCC cells. The proliferation of RCC cells was evaluated using the WST-1 assay and TGFBI expression was detected by real-time PCR (RT-PCR), and Western blot. RESULTS: The results indicated that the expression of TGFBI was significantly decreased by DAC or PTX alone in vitro and in vivo. Moreover, the combination of DAC and PTX caused a synergistic decrease in the expression of TGFBI in RCC cells. We also investigated the effect of VHL-TGFBI signaling on the synergy between DAC and PTX, although the synergy between the two medications was not abolished by interfering with VHL activity or TGFBI expression. RCC cells without VHL activity and RCC cells expressing high levels of TGFBI displayed an increased synergistic effect compared to control cells. CONCLUSION: Our study suggests that VHL-TGFBI signaling is involved in the synergy between DAC and PTX against RCC cells. In addition, the synergy between DAC and PTX is more effective in VHL inactive RCC cells.

VHL-TGFBI signaling is involved in the synergy between 5-aza-2'-deoxycytidine and paclitaxel against human renal cell carcinoma.

PURPOSE: To analyse the role of von Hippel-Lindau (VHL) and transforming growth factor ß-induced (TGFBI) in synergistic mechanisms of 5-aza-2'-deoxycytidine (DAC) and paclitaxel (PTX) against renal cell carcinoma (RCC). METHODS: To elucidate the role in the synergy between DAC and PTX against RCC cells, TGFBI expression was regulated using siRNA technology and an expression vector containing the full-length cDNA for TGFBI was also transfected into RCC cells. The proliferation of RCC cells was evaluated using the WST-1 assay and TGFBI expression was detected by real-time PCR (RT-PCR), and Western blot. RESULTS: The results indicated that the expression of TGFBI was significantly decreased by DAC or PTX alone in vitro and in vivo. Moreover, the combination of DAC and PTX caused a synergistic decrease in the expression of TGFBI in RCC cells. We also investigated the effect of VHL-TGFBI signaling on the synergy between DAC and PTX, although the synergy between the two medications was not abolished by interfering with VHL activity or TGFBI expression. RCC cells without VHL activity and RCC cells expressing high levels of TGFBI displayed an increased synergistic effect compared to control cells. CONCLUSIONS: Our study suggests that VHL-TGFBI signaling is involved in the synergy between DAC and PTX against RCC cells. In addition, the synergy between DAC and PTX is more effective in VHL inactive RCC cells.

Expression and proliferation-promoting role of lymphoid enhancer-binding factor 1 in human clear cell renal carcinoma.

Lymphoid enhancer-binding factor 1 (LEF1) has been regarded as an important gene for carcinogenesis in many malignancies, however, the role of LEF1 in the progression of human renal cell carcinoma (RCC) has not been well studied. In this study, we investigated the expression of LEF1 in human RCC and the effect on proliferative ability of RCC cells. RCC samples from 138 patients who underwent radical nephrectomy were used in this study, the expression of LEF1 protein was determined by immunohistochemistry and Western blot, mRNA expression was analyzed by RT-PCR and real-time PCR. To investigate the effect of LEF1 on the proliferation of RCC cells, a LEF1 vector was transfected into RCC cells and LEF1 expression was also decreased by using siRNA. Proliferative ability of RCC cells was examined by WST-1 assay and a xenograft study with BALB/C nude mice. Our results indicated that LEF1 expression was significantly increased in stage III, IV and grade 3 RCC than in normal kidney, however, decreased LEF1 expression was found in low-stage and grade RCC compared to that in normal kidney, the expression of LEF1 was correlated to tumor stages, histologic grade, and tumor sizes in RCC. The effect of LEF1 on the proliferation in RCC was also analyzed, our results suggested that RCC cells expressing high levels of LEF1 had significantly increased proliferative ability compared to control cell lines, in contrast, RCC cells with a low LEF1 expression had lower proliferative ability. Moreover, LEF1 promoted proliferation of RCC cells depending on suppressing G2/M cell-cycle arrest. Our study demonstrated that the expression of LEF1 is associated with the progression of RCC and that LEF1 maybe involved in the development of RCC, these suggested LEF1 play a key role and might serve as a therapeutic target in treating advanced RCC.

Machine learning algorithms being an auxiliary tool to predict the overall survival of patients with renal cell carcinoma using the SEER database.

Background: The clinical prognosis assessment of renal cell carcinoma (RCC) still relies on nuclear grading and nuclear score by naked eye with microscope, which has defects long time, low efficiency, and uneven evaluation level criteria. There are few machine learning (ML) studies investigating the prognosis in the RCC literature which could also quantify the risk of postoperative recurrence of RCC patients and guide cancer patients to conduct individualized postoperative clinical management. This study evaluated the suitability of ML algorithms for survival prediction in patients with RCC. Methods: A total of 192,912 RCC patients from the Surveillance, Epidemiology, and End Results (SEER) were obtained from 2004 to 2015. Six ML algorithms including support vector machine (SVM), Bayesian method, decision tree, random forest, neural network, and Extreme Gradient Boosting (XGBoost) were applied to predict overall survival (OS) of RCC. Results: Patients from the SEER with a median age of 62 years and the pathological types were clear cell RCC (47.6%), papillary RCC (9.5%), chromophobe RCC (4.0%) and others (4.1%) were collected. In the deleting patients with missing data, the highest accurate model was XGBoost [area under the curve (AUC) 67.0%]. In the deleting patients with missing data and survival time <5 years, the accuracy of random forest, neural network and XGBoost were high, with AUC of 80.8%, 81.5% and 81.8%, respectively. In the only deleting the missing tumor diameter and filling the missing dataset with missForest, the highest accurate model was random forest (AUC: 71.9%). In this study, the overall accuracy of the SVM model was not high, apart from in the population of patients with deleting the missing tumor diameter and survival time <5 years, and filling the missing data with missForest. Random forest, neural network and XGBoost had high accuracy, with AUC of 84.1%, 84.7% and 84.8%, respectively. Conclusions: ML algorithms could be used to predict the prognosis of RCC. It could quantify the recurrence possibility of patients and help more individualized postoperative clinical management. Given the limitations and complexity of datasets, ML may be used as an auxiliary tool to analyze and process larger datasets and complex data.

Gene expression profiling of the synergy of 5-aza-2'-deoxycytidine and paclitaxel against renal cell carcinoma.

BACKGROUND: Renal cell carcinoma (RCC) is one of the most common kidney cancers and is highly resistant to chemotherapy. We previously demonstrated that 5-aza-2'-deoxycytidine (DAC) could significantly increase the susceptibility of renal cell carcinoma (RCC) cells to paclitaxel (PTX) treatment in vitro, and showed the synergy of DAC and PTX against RCC. The purpose of this study is to investigated the gene transcriptional alteration and investigate possible molecular mechanism and pathways implicated in the synergy of DAC and PTX against RCC. METHODS: cDNA microarray was performed and coupled with real-time PCR to identify critical genes in the synergistic mechanism of both agents against RCC cells. Various patterns of gene expression were observed by cluster analysis. IPA software was used to analyze possible biological pathways and to explore the inter-relationships between interesting network genes. RESULTS: We found that lymphoid enhancer-binding factor 1 (LEF1), transforming growth factor ß-induced (TGFBI), C-X-C motif ligand 5 (CXCL5) and myelocytomatosis viral related oncogene (c-myc) may play a pivotal role in the synergy of DAC and PTX. The PI3K/Akt pathway and other pathways associated with cyclins, DNA replication and cell cycle/mitotic regulation were also associated with the synergy of DAC and PTX against RCC. CONCLUSION: The activation of PI3K/Akt-LEF1/ß-catenin pathway could be suppressed synergistically by two agents and that PI3K/Akt-LEF1/ß-catenin pathway is participated in the synergy of two agents.

Persistent Depressive Symptoms and the Changes in Serum Cystatin C Levels in the Elderly: A Longitudinal Cohort Study.

Background: The burden of depression in the elderly is increasing worldwide with global aging. However, there is still a lack of research on the relationship between depressive symptoms and the progression of renal function. Our aim is to evaluate the longitudinal association between baseline depressive symptoms and the changes in serum cystatin C levels over 10 years' follow-up period. Methods: We used longitudinal data from the Health and Retirement Study (HRS), an existing community based nationally representative aging cohort study which enrolled individuals over age 50 in the USA. Depressive symptoms were determined using an eight-item version of the Center for Epidemiologic Studies Depression Scale (CESD) at wave 7 (2004) and wave 8 (2006). Persistent depressive symptoms were defined as both CESD scores measured at waves 7 and 8 were ≥3; episodic depressive symptoms were defined as CESD scores ≥3 at wave 7 or wave 8. A linear mixed model was used to evaluate the correlation between baseline depressive symptoms and future changes in cystatin C levels. Results: The mean age of the 7,642 participants was 63.8 ± 10.8 years, and 60.9% were women. Among the participants, 1,240 (16.2%) had episodic depressive symptoms and 778 (10.2%) had persistent depressive symptoms. Compared with participants with no depressive symptoms at both waves, a significant increase in serum cystatin C levels was found among those with persistent depressive symptoms. Conclusions: Our results showed that baseline persistent depressive symptoms were significantly associated with an increased rate of serum cystatin C levels. The level of serum cystatin C should be monitored in the elderly with persistent depressive symptoms.

Resveratrol Ameliorates Trigeminal Neuralgia-Induced Cognitive Deficits by Regulating Neural Ultrastructural Remodelling and the CREB/BDNF Pathway in Rats.

Chronic pain often leads to cognitive impairment. Resveratrol (Res), a natural polyphenol existing in Polygonum cuspidatum, has been widely investigated for its antinociceptive, anti-inflammatory, and neuroprotective properties. Our aim was to explore the ameliorating effects of resveratrol on pain-related behaviors and learning and memory deficits induced by cobra venom-induced trigeminal neuralgia (TN). The TN model of rats was established by injecting cobra venom solution beneath the epineurium of the infraorbital nerve. Resveratrol was intragastrically administered at a dose of 40 mg/kg twice daily beginning on postoperative day 15. CREB inhibitor 666-15 was intraperitoneally administered at a dose of 10 mg/kg from POD 35-42 after morning resveratrol treatment. Mechanical allodynia was measured via von Frey filaments. Rat free movement was videotaped and analyzed. Spatial learning and memory were evaluated via the Morris water maze test. Ultrastructures of the hippocampal DG region and infraorbital nerve were observed by transmission electron microscopy. We found that resveratrol alleviated TN-induced allodynia, ameliorated learning and memory deficits, restored the ultrastructure of hippocampal neurons and synapses, repaired the damaged myelin sheath of the infraorbital nerve, and activated the CREB/BDNF pathway in the hippocampus of TN rats. CREB inhibitor administration suppressed the resveratrol-rescued abnormal hippocampal ultrastructural changes and aggravated spatial learning and memory impairment by inhibiting CREB/BDNF pathway activation in the hippocampus. Our findings indicated that resveratrol alleviated pain and improved cognitive deficits, probably by regulating neural ultrastructure remodelling and the CREB/BDNF pathway.

Strategy for optical data encryption and decryption using a D-A type stimuli-responsive AIE material.

In this study, we report a new donor-acceptor (D-A) type stimuli-responsive material, (E)-4-(((9-ethyl-9H-carbazol-2-yl)methylene)amino)benzoic acid (C1), which possesses both aggregation-induced emission (AIE) and intramolecular charge transfer (ICT) natures. It glows green photoluminescence which changes into yellow color in response to mechanical stimuli, and fumigation in volatile organic compounds (VOCs) can switch the emission back to the initial state with high reversibility. In addition, the C1 film glows yellow-orange light, but turns into blue emission under continuous fumigation in ethyl acetate vapor. However the vapochromism behaves different when the C1 film is smeared: The emission of the smeared film is similar to the unsmeared but changes into cyan color after fumigation. The differences in vapochromism between smeared and unsmeared film can be easily distinguished by naked eyes. As revealed by SEM, the as-prepared film undergoes a morphology change from ill-shaped particles to microspheres in response to organic vapor, while the smeared film with scratched surface changes into dendritic patterns. According to the morphology study, the vapochromic luminescence can be ascribed to the physical adsorption of ethyl acetate vapor and the resulting change in the ICT process. In light of the unique vapochromism of C1, a new encryption-decryption technique for data recording was developed. Information can be recorded on the C1 film by mechanical writing and simultaneously concealed. It can be only accessed via fumigation in organic vapor, demonstrating a reliable steganography technology.

Treatment of upper urinary tract stones with flexible ureteroscopy in children.

INTRODUCTION: This single-centre, retrospective study aimed to assess the efficacy and safety of flexible ureteroscopy (FURS) combined with holmium laser lithotripsy in treating children with upper urinary tract stones. METHODS: From June 2014 to October 2015, a total of 100 children (74 boys and 26 girls) with upper urinary tract stones were treated using FURS. A 4.7 Fr double-J stent was placed two weeks before operation. Patients were considered stone-free when the absence of residual fragments was observed on imaging studies. The preoperative, operative, and postoperative data of the patients were retrospectively analyzed. RESULTS: A total of 100 pediatric patients with a mean age of 3.51±1.82 years underwent 131 FURS and holmium laser lithotripsy. Mean stone diameter was 1.49±0.92 cm. Average operation time was 30.8 minutes (range 15-60). The laser power was controlled between 18 and 32 W, and the energy maintained between 0.6 and 0.8 J at any time; laser frequency was controlled between 30 and 40 Hz. Complications were observed in 69 (69.0 %) patients and classified according to the Clavien system. Postoperative hematuria (Clavien I) occurred in 64 (64.0 %) patients. Postoperative urinary tract infection with fever (Clavien II) was observed in 8/113 (7.1%) patients. No ureteral perforation and mucosa avulsion occurred. The overall stone-free rate of single operation was 89/100 (89%). Stone diameter and staghorn calculi were significantly associated with stone-free rate. CONCLUSIONS: FURS and holmium laser lithotripsy is effective and safe in treating children with upper urinary tract stones.

Flexible ureteroscopic lithotripsy for the treatment of upper urinary tract calculi in infants.

We evaluated the clinical value of flexible ureteroscopic lithotripsy for the treatment of upper urinary tract calculi in infants. Fifty-five infants with upper urinary tract calculi were included in this study: 41 males and 14 females. Retrograde intrarenal surgery was performed by an 8 Fr/30 cm flexible ureterorenoscope (POLY®) combined with a holmium laser. CT scanning or radiography of the kidneys, ureters, and bladder region was performed one month after the operation to confirm the clearance of calculi. All the 55 infants with calculi in 74 sides underwent 66 flexible ureteroscopic lithotripsy procedures. The median operation time was 30 min. The median amount of flushing fluid was 500 mL. The stone-free rate after a single session treatment was 94.6%, within which 10 infants underwent simultaneous bilateral flexible ureteroscopy lithotripsy. Catheters were retained in 45 infants for 24-48 h after the operation. Continuous high fever due to reflux was present in two cases. Flushing fluid extravasation was found in one infant. Some patients with minor complications, such as mild hematuria, irritation symptoms, and low fever, recovered without treatment. The duration of hospitalization time after the operation was approximately 1-5 days. Flexible ureteroscopic lithotripsy is a safe, highly efficient, minimally invasive, and reproducible operation for removal of upper urinary tract calculi in infants. This technique is a convenient method for postoperative management of patients that enhances their rapid recovery. It is a promising option for therapy of infants ineffectively treated by extracorporeal shockwave lithotripsy.

Decitabine induces G2/M cell cycle arrest by suppressing p38/NF-κB signaling in human renal clear cell carcinoma.

OBJECTIVE: The anti-neoplastic effects of decitabine, an inhibitor of DNA promoter methylation, are beneficial for the treatment of renal cell carcinoma (RCC); however, the mechanism of action of decitabine is unclear. We analyzed gene expression profiling and identified specific pathways altered by decitabine in RCC cells. METHODS: Four human RCC cell lines (ACHN, Caki-1, Caki-1, and A498) were used in this study; growth suppression of RCC cells by decitabine was analyzed using the WST-1 assay. Apoptosis and cell cycle arrest were examined using flow cytometric analysis. Gene expression of RCC cells induced by decitabine was evaluated with cDNA microarray, and potential biological pathways were selected using Ingenuity Pathway Analysis. The activity of the p38-NF-κB pathway regulated by decitabine was confirmed by Western blotting. RESULTS: Decitabine suppresses the proliferation of RCC cells in vitro. Although decitabine did not significantly induce apoptosis, decitabine caused cell cycle arrest at G2/M in a dose-dependent manner. Gene expression regulated by decitabine in RCC cells was investigated using microarray analysis. Ubiquitin carboxyl terminal hydrolase 1 (UCHL1), interferon inducible protein 27 (IFI27), and cell division cycle-associated 2 (CDCA2) may be involved in growth suppression of RCC cells by decitabine. The phosphorylation of p38-NF-κB pathway was suppressed by decitabine in RCC cells. CONCLUSIONS: We investigated gene expression profiling and pathways modulated by decitabine in RCC cells. Decitabine was shown to suppress the growth of RCC cells via G2/M cell cycle arrest and the p38-NF-κB signaling pathway may play a role in the anti-neoplastic effect of decitabine in RCC cells.

Interferon-α enhances the susceptibility of renal cell carcinoma to rapamycin by suppressing mTOR activity.

The aim of the present study was to investigate the antiproliferative effects of interferon (IFN)-α and rapamycin (RPM) on renal cell carcinoma (RCC) cells and examine the synergistic growth suppression conferred by IFN-α and RPM. The effects of IFN-α and/or RPM on RCC cells were determined using a WST-1 assay and the synergy of IFN-α and RPM against three RCC cell lines was analyzed with isobolographic analysis. The expression of mammalian target of rapamycin (mTOR) was downregulated by RNAi, and the expression and phosphorylation of proteins in the mTOR pathway following treatment with IFN-α and/or RPM was examined by western blot analysis. The observations indicated that IFN-α significantly increased the susceptibility of RCC cells to RPM and the synergistic effect of IFN-α and RPM against RCC cells was confirmed in all three RCC cell lines. The mTOR pathway was shown to be associated with the synergistic effect of IFN-α and RPM against RCC. IFN-α and RPM alone decreased the phosphorylation of mTOR, p70 S6 kinase, S6 and 4E binding protein 1, and IFN-α significantly enhanced the RPM-induced suppression of the mTOR pathway. However, in RCC cells with low mTOR activity, the synergy of IFN-α and RPM was eliminated. Therefore, the results of the present study indicate that the mTOR pathway plays an important role in the synergistic effect of IFN-α and RPM against RCC cells. Thus, mTOR may serve as an effective therapeutic target in the treatment of advanced RCC.

5-aza-2'-deoxycytidine enhances susceptibility of renal cell carcinoma to paclitaxel by decreasing LEF1/phospho-ß-catenin expression.

We investigated the molecular mechanisms by which 5-aza-2'-deoxycytidine (DAC) and paclitaxel (PTX) use lymphoid enhancer-binding factor 1 (LEF1) and the Wnt/ß-catenin pathway to synergistically interact against renal cell carcinoma (RCC). LEF1 expression was examined by real-time PCR and immunohistochemistry. The regulation of LEF1/ß-catenin protein expression by DAC and/or PTX was examined by Western blot and immunoprecipitation. To analyze the effect of LEF1 on the proliferative ability of RCC cells and the synergy of DAC and PTX against RCC cells, an expression vector containing the full-length cDNA for LEF1 was transfected into RCC cells, and LEF1 expression was also decreased using siRNA technology. Our results confirmed that DAC and PTX synergistically decreased the expression of LEF1 in vivo and in vitro. Moreover, treatment of RCC cell lines with the combination of DAC and PTX caused a synergistic decrease in LEF1/phospho-ß-catenin. Our study also demonstrated a negative correlation between LEF1 expression and the proliferative ability of RCC cells. Although interfering with LEF1 expression did not abolish the synergy between the two agents, RCC cells expressing high levels of LEF1 displayed an increased synergistic effect compared with RCC cells expressing low levels of LEF1. This study suggests that LEF1 can enhance the proliferation of RCC cells and that the LEF1/ß-catenin complex plays an important role in the synergy of DAC and PTX against RCC cells. Moreover, the synergy between DAC and PTX may be more effective in RCC cells expressing high levels of LEF1.