Needle-perc-assisted endoscopic surgery versus retrograde intrarenal surgery for the treatment of 1- to 2-cm lower-pole renal stones in patients with unfavorable infundibulopelvic anatomy: a matched-pair analysis.

PURPOSE: To compare the safety and efficacy of needle-perc-assisted endoscopic surgery (NAES) and retrograde intrarenal surgery (RIRS) for the treatment of 1- to 2-cm lower-pole stones (LPS) in patients with complex infundibulopelvic anatomy. METHODS: Between June 2020 and July 2022, 32 patients with 1- to 2-cm LPS and unfavorable lower-pole anatomy for flexible ureteroscopy were treated with NAES. The outcomes of these patients were compared with patients who underwent RIRS using matched-pair analysis (1:1 scenario). The matching parameters such as age, gender, body mass index, stone size, hardness, and pelvicalyceal anatomy characteristics including infundibular pelvic angle, infundibular length, and width were recorded. Data were analyzed using the Student's t-test, Mann-Whitney U test, and Fisher's exact test. RESULTS: The two groups had similar baseline characteristics and lower-pole anatomy. The stone burden was comparable between both groups. NASE achieved a significantly better initial stone-free rate (SFR) than RIRS (87.5% vs 62.5%, p = 0.04). The auxiliary rates for the NAES and RIRS groups were 12.5% and 31.3%, respectively (p = 0.13). Finally, the SFR after 1 month follow-up period was still higher for the NAES group than RIRS group (93.8% versus 81.3%), but the difference was not statistically significant (p = 0.26). Concerning the operation duration, overall complication rates, and postoperative hospital stay, there were no differences between two groups. CONCLUSION: Compared to RIRS for treating 1- to 2-cm LPS in patients with unfavorable infundibulopelvic anatomy for flexible ureteroscopy, NAES was safe and effective with higher SFR and similar complication rate.

Ultrasound-guided renal access and balloon dilation for PCNL in the prone position: results of a multicenter prospective observational study.

PURPOSE: To investigate the safety and efficacy of ultrasound-guided renal access and tract dilation using balloon dilators, as well as to identify suitable patients for this technique. METHODS: Consecutive patients undergoing ultrasound-guided PCNL using balloon dilators between December 2019 and June 2020 in seven large medical centers from China were prospectively enrolled. Demographic and perioperative parameters of the patients were collected. Logistic regression analysis was used to analyze factors that would affect the success rate of tract establishment using ultrasound-guided renal access and balloon dilation. RESULTS: A total of 170 patients were included in this study, among whom, 91.18% of the (155/170) patients had a successful tract establishment under ultrasound guidance on the first attempt. The stone-free rate was 83.5% and postoperative complications occurred in 14 patients (8.23%). In univariate analysis, history of ipsilateral surgery (p = 0.026), and stone diameter (p = 0.01) were significantly associated with tract establishment failure, while a larger width of the target calyx (p = 0.016) and the presence of hydronephrosis (p = 0.001) were significantly associated with a successful tract establishment. In multivariate analysis, only hydronephrosis in target calyx (p = 0.027) was a favorable factor for successful tract establishment, and the history of ipsilateral renal surgery (p = 0.012) was the only independent risk factor for failure of tract establishment. CONCLUSION: It was safe and effective to establish percutaneous renal access with balloon dilation under whole-process ultrasound monitoring during PCNL. Furthermore, patients with a hydronephrotic target calyx and without history of ipsilateral renal surgery were most suited to this technique. Trial registration CHiCTR1800014448.

Needle-perc-assisted endoscopic surgery for patients with complex renal stones: technique and outcomes.

Our aim was to investigate the safety and efficacy of needle-perc-assisted percutaneous nephrolithotomy (PCNL) or retrograde intrarenal surgery (RIRS), namely, needle-perc-assisted endoscopic surgery (NAES), in a series of patients with large and/or complex renal stones. From May 2018 to August 2021, a total of 119 patients underwent NAES at our institute. Among them, 94 patients underwent needle-perc-assisted standard PCNL in prone position and 25 underwent needle-perc-assisted RIRS in the Galdakao-modified supine Valdivia position or prone split-leg position. Clinical factors including age, sex, medical history, and stone characteristics were collected. Intraoperative and postoperative outcomes were retrospectively evaluated. The patients' mean age ± standard deviation was 50.3 ± 14.3 years. The mean stone size was 7.6 ± 3.7 and 1.7 ± 0.8 cm for needle-perc-assisted PCNL and RIRS, respectively. Of the 119 patients, 51 had staghorn stones, 16 had solitary kidneys, 17 had a history of ipsilateral renal surgery, and 6 had calyceal diverticular stones. The mean operative time was 83.4 ± 25.9 min for needle-perc-assisted PCNL and 66.3 ± 21.8 min for needle-perc-assisted RIRS. The stone-free rate (SFR) for needle-perc-assisted PCNL was 77.7% after the first treatment and 88.3% after auxiliary treatments. The SFR for needle-perc-assisted RIRS was 88.0% and no auxiliary treatments were carried out in this group. Eleven (11.7%) patients who underwent needle-perc-assisted standard PCNL developed Clavien-Dindo grade I or II complications. Three (12.0%) patients who underwent needle-perc-assisted RIRS developed a fever (grade I). The overall complication rate for NAES was 11.8%, with no urosepsis, angioembolization, or other grade III to V complications. In conclusion, NAES is a safe and effective procedure for one-step complete resolution of large and/or complex renal stones with no additional procedure-related complications.

Outcomes of ultrasound-guided percutaneous nephrolithotomy for the treatment of large stones within non-functioning atrophic kidneys.

OBJECTIVES: To evaluate outcomes in patients who underwent total ultrasound-guided percutaneous nephrolithotomy for the management of large stones (>2 cm) within non-functioning atrophic kidneys, and to identify risk factors associated with postoperative persistence of urinary tract infection symptoms in such patients. METHODS: Between December 2014 and May 2019, 56 patients with large stones within non-functioning atrophic kidneys were treated at Beijing Tsinghua Changgung Hospital, Beijing, China, with total ultrasound-guided percutaneous nephrolithotomy. Factors including age, sex, previous medical history, stone burden, stone composition, stone-free rate, complication rate and postoperative urinary tract infection symptoms were retrospectively evaluated. Uni- and multivariate analyses were carried out to identify risk factors that predict persistence of urinary tract infection symptoms after percutaneous nephrolithotomy in such patients. RESULTS: The final stone-free rate after auxiliary treatments was 87.5%. The overall complication rate was 17.9%. After a median follow-up period of 12 months (range 6-40 months), three patients (5.4%) ultimately underwent nephrectomy. A total of 44 patients (78.6%) had no symptoms of urinary tract infection during follow up, whereas 63.6% of patients with diabetes continued to show persistent urinary tract infection symptoms at the latest follow up. Five patients (8.9%) showed improved total renal function, while most patients (83.9%) showed a stable estimated glomerular filtration rate. Diabetes was the only factor showing statistical significance in both univariate and multivariate analyses that predicted persistence of urinary tract infection symptoms after treatment with percutaneous nephrolithotomy. CONCLUSIONS: Ultrasound-guided percutaneous nephrolithotomy is a safe and feasible procedure that can be carried out in patients with large stones within non-functioning atrophic kidneys. However, for such patients with diabetes, percutaneous nephrolithotomy is not recommended.

Let-7d inhibits intratumoral macrophage M2 polarization and subsequent tumor angiogenesis by targeting IL-13 and IL-10.

The microRNA let-7d has been reported to be a tumor suppressor in renal cell carcinoma (RCC). Tumor-associated macrophages (TAM) are M2-polarized macrophages that can enhance tumor growth and angiogenesis in many human cancers. However, the role of let-7d in TAM-associated RCC progression remains elusive. First, we observed a strongly inverse correlation between let-7d expression and microvessel density in RCC tissues. Furthermore, the proliferation, migration, and tube formation of HUVECs were significantly inhibited by conditioned medium from a coculture system of the phorbol myristate acetate pretreated human THP-1 macrophages and let-7d-overexpressing RCC cells. Moreover, the proportion of M2 macrophages was significantly lower in the group that was cocultured with let-7d-overexpressing RCC cells. Subcutaneous xenografts formed by the injection of let-7d-overexpressing RCC cells together with THP-1 cells resulted in a significant decrease in the M2 macrophage ratio and microvessel density compared with those formed by the injection of control RCC cells with THP-1 cells. In silico and experimental analysis revealed interleukin-10 (IL-10) and IL-13 as let-7d target genes. Importantly, the addition of IL-10 and IL-13 counteracted the inhibitory effects of the conditioned medium from the coculture system with let-7d-overexpressing RCC cells in vitro. Additionally, overexpression of IL-10 and IL-13 reversed the effects of let-7d on macrophage M2 polarization and tumor angiogenesis in vivo. Finally, the expression of IL-10 and IL-13 were inversely correlated with the expression of let-7d in RCC clinical specimens. These results suggest that let-7d may inhibit intratumoral macrophage M2 polarization and subsequent tumor angiogenesis by targeting IL-10 and IL-13.

A Novel Surgical Technique for Treatment of Renal Stones in Preschool-aged Patients: Initial Experience With Needle-perc.

OBJECTIVE: To describe and evaluate our newly developed minimal instrumentation technique, needle-perc, for treatment of preschool-aged patients with renal stones. To the best of our knowledge, this is the smallest endoscopic equipment for percutaneous nephrolithotomy reported thus far. MATERIALS AND METHODS: Needle-perc was performed in 8 patients using a 4.2-Fr needle to achieve access to the collecting system under ultrasonic guidance alone. The mean patient age was 2.4 years (range, 0-5 years). Five of the 8 patients were boys, while 3 were girls. Six patients had unilateral stones and 2 patients had bilateral stones. Six renal units exhibited single calculus, 2 exhibited staghorn stones, and 2 exhibited multiple stones. The mean calculus size was 1.6 cm (range, 0.8-4.5 cm). Preoperative, intraoperative, and postoperative parameters were analyzed prospectively. RESULTS: Access was successfully achieved in all patients. Puncturing was fully guided by ultrasound. Five patients underwent needle-perc alone. Two patients were transferred to 16-Fr access because of intrarenal infection and large stone burden. One patient underwent reduction of tract number. The mean operative time was 49.2 minutes (range, 22-75 minutes); mean hemoglobin loss was 5.2 g/L (range, 0-13.8 g/L). The mean postoperative hospital stay was 5 days (range, 3-7 days). Preplaced catheters were kept for 2-3 days to facilitate fragment discharge. Complications occurred in 2 patients: fever >38.5°C. The stone-free rate was 100% at 1 month postoperatively. CONCLUSION: Our initial data indicate that needle-perc is feasible and safe for preschool-aged renal patients. Further studies are required to define its usefulness in treatment of larger stones.

MiR-21 promotes calcium oxalate-induced renal tubular cell injury by targeting PPARA.

Kidney stone disease is a crystal concretion formed in the kidneys that has been associated with an increased risk of chronic kidney disease. MicroRNAs are functionally involved in kidney injury. Data mining using a microRNA array database suggested that miR-21 may be associated with calcium oxalate monohydrate (COM)-induced renal tubular cell injury. Here, we confirmed that COM exposure significantly upregulated miR-21 expression, inhibited proliferation, promoted apoptosis, and caused lipid accumulation in an immortalized renal tubular cell line (HK-2). Moreover, inhibition of miR-21 enhanced proliferation and decreased apoptosis and lipid accumulation in HK-2 cells upon COM exposure. In a glyoxylate-induced mouse model of renal calcium oxalate deposition, increased miR-21 expression, lipid accumulation, and kidney injury were also observed. In silico analysis and subsequent experimental validation confirmed the peroxisome proliferator-activated receptor (PPAR)-α gene (PPARA) a key gene in fatty acid oxidation, as a direct miR-21 target. Suppression of miR-21 by miRNA antagomiR or activation of PPAR-α by its selective agonist fenofibrate significantly reduced renal lipid accumulation and protected against renal injury in vivo. In addition, miR-21 was significantly increased in urine samples from patients with calcium oxalate renal stones compared with healthy volunteers. In situ hybridization of biopsy samples from patients with nephrocalcinosis revealed that miR-21 was also significantly upregulated compared with normal kidney tissues from patients with renal cell carcinoma who underwent radical nephrectomy. These results suggested that miR-21 promoted calcium oxalate-induced renal tubular cell injury by targeting PPARA, indicating that miR-21 could be a potential therapeutic target and biomarker for nephrolithiasis.

MicroRNA-200b is downregulated and suppresses metastasis by targeting LAMA4 in renal cell carcinoma.

BACKGROUND: Metastasis is the primary cause of tumor death in renal cell carcinoma (RCC). Improved diagnostic markers of metastasis are critically needed for RCC. MicoRNAs are demonstrated to be stable and significant biomarkers for several malignancies. In this study, we aimed to explore the metastasis related microRNAs and its mechanism in RCC. METHODS: The relationship between microRNAs expression and prognosis and metastasis of RCC patients were explored by data mining through expression profiles from The Cancer Genome Atlas (TCGA). A total of 80 RCC tissues and adjacent normal kidney tissues were obtained from Department of Urology, Peking University First Hospital. Expression of microRNA-200b (miR-200b) in RCC tissues and cell lines were determined by bioinformatic data mining and quantitative real-time PCR (qRT-PCR). The effects of miR-200b on cell proliferation, migration and invasion were determined by cell counting kit-8 and colony formation assay, wound healing assay and Boyden chamber assay. Mouse cell-derived xenograft and patient-derived xenograft model were also performed to evaluate the effects of miR-200b on tumor growth and metastasis in vivo. The molecular mechanism of miR-200b function was investigated using bioinformatic target predication and high-throughput cDNA sequencing (RNA-seq) and validated by luciferase reporter assay, qRT-PCR, Western blot and immunostaining in vitro and in vivo. FINDINGS: Our findings indicates that miR-200b is frequently downregulated and have potential utility as a biomarker of metastasis and prognosis in RCC. Interestingly, ectopic expression of miR-200b in the Caki-1 and OSRC-2 cell lines suppresses cell migration and invasion in vitro as well as tumor metastases in vivo. However, miR-200b has no effect on cell proliferation in vitro and tumor growth in vivo. In addition, bioinformatics target predication and RNA-seq results reveals that Laminin subunit alpha 4 (LAMA4) is one target of miR-200b and significantly inhibited by miR-200b in vitro and in vivo. INTERPRETATION: These results demonstrate a previously undescribed role of miR-200b as a suppressor of tumor metastasis in RCC by directly destabilizing LAMA4 mRNA.

Long-term outcomes of ultrasound-guided percutaneous nephrolithotomy in patients with solitary kidneys: a single-center experience.

PURPOSE: To report our experience with total ultrasound-guided percutaneous nephrolithotomy (PCNL) in the management of patients with solitary kidney, and evaluate the safety and feasibility of this technique. MATERIALS AND METHODS: Between October 2014 and December 2016, 48 patients with solitary kidneys underwent total ultrasound-guided PCNL at our institution. Stone-free rate (SFR), auxiliary procedures, and complications were recorded. Changes in renal function were evaluated by comparing preoperative and postoperative estimated glomerular filtration rates (eGFRs). Perioperative factors that may affect renal function were analyzed to define factors predicting renal function improvement on long-term follow-up. Of 48 patients, 44 were followed at least 6 months, whereas four patients were lost to follow-up. RESULTS: Among all patients, staghorn calculi were found in 18 (37.5%) patients. 14 (29.2%) patients required a two-stage PCNL. Struvite was found in six (12.5%) patients. Complications were reported in eight (16.7%) patients. Severe bleeding was noticed in three patients; no angioembolization was required. After a median follow-up of 12 (6-26) months, the final SFR was 81.8% after auxiliary treatments. There was a significant improvement of eGFR from 53.9 ± 24.0 to 61.3 ± 25.4 mL/min/1.73 m2 (P < 0.01). Renal function was stable, improved and worse in 65.9% (n = 29), 27.3% (n = 12), and 6.8% (n = 3) of patients, respectively, compared with preoperative levels. CONCLUSIONS: Ultrasound-guided PCNL is a safe and feasible procedure with an acceptably low complication rate in patients with solitary kidneys. At long-term follow-up, the renal function in more than 90% of the patients with solitary kidneys can be improved or stabilized after ultrasound-guided PCNL.

Simultaneous antegrade and retrograde endoscopic treatment of non-malignant ureterointestinal anastomotic strictures following urinary diversion.

BACKGROUND: The ureterointestinal anastomosis stricture (UAS) is a common complication of urinary diversion after radical cystectomy. For decades, open anastomotic revision remained the gold standard for the treatment of UAS. However, with the advancement in endoscopic technology, mini-invasive therapeutic approaches have been used in its management. Here, we report our experience with and long-term results of combined simultaneous antegrade and retrograde endoscopy (SARE) in the treatment of non-malignant UASs after urinary diversion in a consecutive series of patients. METHODS: From March 2012 to January 2015, there were 32 consecutive patients with 32 non-malignant UASs following radical cystectomy and urinary diversion. Twenty-nine patients were treated with SARE technique and comprised the study group. Using simultaneous antegrade flexible ureteroscope combined with retrograde semi-rigid ureteroscope or nephroscope, partial or complete strictures were managed with laser incision and balloon dilation under direct visualization. A 7/12 Fr graded endopyelotomy stent was left for 3-6 months after the procedure. Success was defined as symptomatic improvement and radiographic resolution of obstruction. RESULTS: With a median followup of 22 months (6-36), the overall success rate for SARE was 69.0%. Twenty patients with partial stricture had a success rate of 85%, and 9 patients with complete stricture had a success rate of 33.3%. Renal function, hydronephrosis grade, stricture type, and stricture length were significant influences on the outcome (P < 0.05). No complication was observed. CONCLUSIONS: The SARE is a safe and effective treatment for UAS, and may be the only endoscopic treatment approach for complete UAS. While success rate for complete strictures is low compared to open revision, it should be considered as an initial approach given its low overall morbidity. For partial strictures, prudent patient selection results in higher success rates that are nearly comparable to open revision.

High expression of KPNA2 defines poor prognosis in patients with upper tract urothelial carcinoma treated with radical nephroureterectomy.

BACKGROUND: To analyze the expression of karyopherin alpha 2 (KPNA2) in upper tract urothelial carcinoma (UTUC) and to investigate whether the KPNA2 expression provides additional prognostic information following radical nephroureterectomy (RNU). METHODS: A tissue microarray (TMA) containing samples from 176 patients with UTUC who underwent RNU at our institute was analyzed for KPNA2 expression using immunohistochemistry. KPNA2 expression in normal urothelial cell line and urothelial carcinoma cell lines was evaluated by western blot analysis. Using RNA interference in vitro, the effects of KPNA2 inhibition on cellular viability, migration and apoptosis were determined. RESULTS: KPNA2 expression was significantly upregulated in the UTUC samples compared with the adjacent normal urothelial tissues. High KPNA2 immunoreactivity was identified as a predictor of bladder recurrence (hazard ratio [HR]: 2.017, 95% CI 1.13-3.61, p = 0.018), poor disease-free survival (DFS, HR: 2.754, 95% CI 1.68-4.51, p = 0.001) and poor overall survival (OS, HR: 4.480, 95% CI 1.84-10.89, p = 0.001) for patients with UTUC after RNU. Furthermore, high KPNA2 immunoreactivity was independent of the conventional predictive factors in a multivariate analysis. Additional in vitro experiments revealed that KPNA2 expression was higher in urothelial carcinoma cell lines than in normal urothelial cell line. KPNA2 inhibition with a specific siRNA decreased cell viability and migration and increased apoptosis in urothelial carcinoma cell lines. CONCLUSIONS: KPNA2 is a novel independent prognostic marker for bladder recurrence, DFS and OS of UTUC patients who have undergone RNU. Moreover, these data suggest that KPNA2 may be a promising therapeutic target for UTUC.

Human adipose derived stem cells induced cell apoptosis and s phase arrest in bladder tumor.

The aim of this study was to determine the effect of human adipose derived stem cells (ADSCs) on the viability and apoptosis of human bladder cancer cells. EJ and T24 cells were cocultured with ADSCs or cultured with conditioned medium of ADSCs (ADSC-CM), respectively. The cell counting and colony formation assay showed ADSCs inhibited the proliferation of EJ and T24 cells. Cell viability assessment revealed that the secretions of ADSCs, in the form of conditioned medium, were able to decrease cancer cell viability. Wound-healing assay suggested ADSC-CM suppressed migration of T24 and EJ cells. Moreover, the results of the flow cytometry indicated that ADSC-CM was capable of inducing apoptosis of T24 cells and inducing S phase cell cycle arrest. Western blot revealed ADSC-CM increased the expression of cleaved caspase-3 and cleaved PARP, indicating that ADSC-CM induced apoptosis in a caspase-dependent way. PTEN/PI3K/Akt pathway and Bcl-2 family proteins were involved in the mechanism of this reaction. Our study indicated that ADSCs may provide a promising and practicable manner for bladder tumor therapy.

Expression of oncogenic HMGN5 increases the sensitivity of prostate cancer cells to gemcitabine.

Prostate cancer is a leading cause of cancer-related death among men. Early diagnosis and treatment are successful against prostate cancer, yet the clinical treatment of advanced prostate cancer remains a challenge. Gemcitabine is used to treat a broad spectrum of solid tumors; however, the clinical response of prostate cancer patients to gemcitabine is limited. In the present study, we showed that HMGN5, a nucleosome binding protein that can unfold chromatin by binding to histone (H1), is overexpressed in prostate cancer cells and plays an oncogenic role in prostate cancer tumorigenesis and development by activating the MAPK signaling pathway. We also found that sensitivity of prostate cancer cells to gemcitabine was positively correlated with HMGN5 expression. Knockdown of HMGN5 expression reduced the sensitivity of PC-3 cells to gemcitabine, and ectopic HMGN5 expression in DU145 cells enhanced the sensitivity to gemcitabine. Gemcitabine decreased HMGN5 expression, consequently leading to inactivation of the MAPK signaling pathway and cleavage of the PARP protein. Finally, we showed that PC-3 cells acquire gemcitabine resistance by gradual loss of HMGN5 expression. The present study suggests that HMGN5 is a potential biomarker for treating prostate cancer, and patients with a high level HMGN5 will benefit from gemcitabine treatment.

HMGN5 knockdown sensitizes prostate cancer cells to ionizing radiation.

BACKGROUND: High Mobility Group N (HMGN) proteins are a family of chromatin structural proteins that specifically bind to nucleosome core particles. HMGN5 is a novel and characteristic member of the HMGN protein family. We have previously found that HMGN5 is upregulated in prostate cancer and its downregulation had been demonstrated to induce apoptosis and G2-M cell cycle arrest. METHODS: The radiosensitization effect of HMGN5 knockdown on PC3 and DU145 cells was assessed using clonogenic assay, flow cytometry, and comet assay. The DNA double-strand break (DSB) repair kinetics of HMGN5 knockdown and control cells after radiation exposure was evaluated using immunocytofluorescence. The mitochondrial reactive oxygen species (ROS) levels were estimated using Dihydrorhodamine 123 (DHR 123) probes. Expression of mitochondrial antioxidant MnSOD was measured by real-time PCR and Western blot. The expression of antiapoptotic proteins Bcl-2 and Bcl-xL as well as cleavage of caspase-3, caspase-9, and PARP were also measured using Western blot. RESULTS: HMGN5 knockdown cells exhibit decreased clonogenic survival and increased apoptosis rate in response to 2-8 Gy ionizing radiation (IR). Loss of HMGN5 does not affect the DSB repair kinetics after radiation exposure. HMGN5 knockdown cells demonstrated increased mitochondrial ROS level and suppressed induction of MnSOD upon radiation compared with control cells upon radiation. Further, MnSOD knockdown resulted in inhibited cell viability as well as increased mitochondrial ROS level and apoptosis upon radiation in PC3 and DU145 cells. Finally, HMGN5 knockdown cells showed significantly decreased levels of antiapoptotic proteins Bcl-2 and Bcl-xL as well as increased cleavage of caspase-3, caspase-9, and PARP compared with control cells after radiation. CONCLUSIONS: HMGN5 knockdown sensitizes prostate cancer cells to ionizing radiation, and the radiosensitization effect may be partially mediated through suppressed induction of MnSOD and enhanced activation of apoptosis pathway in response to IR.

KCNJ1 inhibits tumor proliferation and metastasis and is a prognostic factor in clear cell renal cell carcinoma.

Potassium inwardly rectifying channel, subfamily J, member 1 (KCNJ1), as an ATP-dependent potassium channel, plays an essential role in potassium balance. KCNJ1 variation is associated with multiple diseases, such as antenatal Bartter syndrome and diabetes. However, the role of KCNJ1 in clear cell renal cell carcinoma (ccRCC) is still unknown. Here, we studied the expression and function of KCNJ1 in ccRCC. The expression of KCNJ1 was evaluated in ccRCC tissues and cell lines by quantitative real-time PCR (qRT-PCR), Western blot, and immunohistochemistry analysis. The relationship between KCNJ1 expression and clinicopathological characteristics was analyzed. p3xFLAG-CMV-14 vector containing KCNJ1 was constructed and used for transfecting ccRCC cell lines 786-O and Caki-2. The effects of KCNJ1 on cell proliferation, invasion, and apoptosis were detected in ccRCC cell lines using cell proliferation assay, transwell assay, and flow cytometry, respectively. We found that KCNJ1 was low-expressed in ccRCC tissues samples and cell lines, and its expression level was significantly associated with tumor pathology grade (P = 0.002) and clinical stage (P = 0.023). Furthermore, the KCNJ1 expression was a prognostic factor of ccRCC patient's survival (P = 0.033). The re-expression of KCNJ1 in 786-O and Caki-2 significantly inhibited cancer cell growth and invasion and promoted cancer cell apoptosis. Moreover, knockdown of KCNJ1 in HK-2 cells promoted cell proliferation. Collectively, these data highlight that KCNJ1, low-expressed in ccRCC and associated with poor prognosis, plays an important role in ccRCC cell growth and metastasis.

Let-7d suppresses growth, metastasis, and tumor macrophage infiltration in renal cell carcinoma by targeting COL3A1 and CCL7.

BACKGROUND: MicroRNAs are endogenous small noncoding RNAs that are functionally involved in numerous critical cellular processes including tumorigenesis. Data mining using a microRNA array database suggested that let-7d microRNA may be associated with renal cell carcinoma (RCC) malignant progression. Here, we performed further analyses to determine whether let-7d is functionally linked to RCC malignancy. METHODS: Quantitative real-time PCR was used to determine the level of mature let-7d in RCC clinical specimens and its correlation with clinicopathological data. Immunohistochemical staining was conducted to characterize the stroma of RCC. Let-7d overexpressing RCC cell lines combined with mouse models bearing cell-derived xenografts and patient-derived xenografts were used to assess the functional role of let-7d in vitro and in vivo. RESULTS: Downregulation of let-7d in clinical RCC samples was associated with advanced tumor grade and T stage and increased vascular invasion. An inverse relationship between let-7d expression and macrophage infiltration was found in clinical RCC samples. Functional studies indicated that ectopic expression of let-7d significantly inhibited RCC cell proliferation, migration, and peripheral blood monocyte (PBMC) recruitment in vitro, as well as tumor growth, metastasis, and tumor macrophage infiltration in vivo. In silico analysis and subsequent experimental validation confirmed collagen, type III, alpha 1 (COL3A1) and C-C subfamily chemokine member CCL7 as direct let-7d target genes. The addition of COL3A1 and CCL7 counteracted the inhibitory effects of let-7d on RCC cell proliferation, migration, and PBMC recruitment. The inhibition of let-7d increased cell proliferation, migration, and PBMC recruitment by the enhanced expression of COL3A1 and CCL7 genes in vitro. The mRNA levels of COL3A1 and CCL7 were inversely correlated with let-7d level in RCC clinical specimens. CONCLUSIONS: These results suggest that let-7d may suppress RCC growth, metastasis, and tumor macrophage infiltration at least partially through targeting COL3A1 and CCL7.

Resiniferatoxin for treatment of lifelong premature ejaculation: a preliminary study.

OBJECTIVES: To evaluate the efficacy of resiniferatoxin in the treatment of patients with lifelong premature ejaculation. METHODS: A total of 41 outpatients (mean age 26.14 ± 4 years) with premature ejaculation completed the present study. They were randomly separated into the resiniferatoxin group and the placebo group. The resiniferatoxin group included 11 patients with redundant prepuce and 10 patients without redundant prepuce, whereas the placebo group contained 10 patients with redundant prepuce and 10 patients without. For the treatment, the glans were respectively soaked in 30 mL of resiniferatoxin with a concentration of 100 nmol/L or 10% alcohol solution for 30 min before sexual intercourse. Clinical efficacy was assessed by using the Chinese Index of Sexual Function for Premature Ejaculation-5 and the intravaginal ejaculation latency time before or 4 weeks after the treatment. The side-effects were also evaluated. RESULTS: In the resiniferatoxin group, the effective rate of patients with redundant prepuce was 63.6%, and both the intravaginal ejaculation latency time and Chinese Index of Sexual Function for Premature Ejaculation-5 significantly increased (P < 0.05). However, the effective rate of patients without redundant prepuce was 20%, and there were no significant changes of their intravaginal ejaculation latency time and Chinese Index of Sexual Function for Premature Ejaculation-5 before and after the resiniferatoxin treatment (P > 0.05). The total effective rate of patients treated with resiniferatoxin was 42.9%. In the placebo group, the effective rate of patients with or without redundant prepuce was 20% and 10%, respectively. The total effective rate of patients treated with placebo was 15%, and there were no significant changes of their intravaginal ejaculation latency time and Chinese Index of Sexual Function for Premature Ejaculation-5 before and after the placebo treatment (P > 0.05). The side-effects included a slight burning sensation for the glans penis and dysuria. CONCLUSIONS: These preliminary results show that resiniferatoxin might be suitable for treating patients with lifelong premature ejaculation and particularly those with redundant prepuce.