Hypertension Resolution after Laparoscopic Adrenal Tumor Resection in Patients of Adrenal Incidentaloma with Normal Hormone Levels.

INTRODUCTION: Postoperative hypertension resolution among patients with adrenal incidentalomas and normal hormone levels was unknown. Identifying the predictive factors was beneficial to the management of adrenal incidentalomas. METHODS: We conducted a retrospective cohort study, recruiting patients undergoing laparoscopic adrenal tumor resection for adrenal incidentaloma with hypertension and normal hormone levels. Demographic, clinical, treatment, and laboratory data were collected and compared. We used univariable and multivariable logistic regression methods to identify the predictive factors of postoperative hypertension resolution. RESULTS: Of the 171 patients in our study, 130 (76.0%) patients performed a resolution of hypertension, and 57 (33.3%) patients had a significant reduction. Multivariate logistic regression analysis showed that the male sex (odds ratio (OR) 0.305, 95% confidence interval (CI): 0.098-0.948, p = 0.040), body mass index (BMI) (OR 0.973, 95% CI: 0.670-0.938, p = 0.007), aldosterone and plasma renin activity ratio (APR) in erect position (OR 1.206, 95% CI: 1.042-1.397, p = 0.012), and preoperative systolic pressure (OR 1.044, 95% CI: 1.009-1.080, p = 0.014), were significantly associated with the outcomes of hypertension resolution. DISCUSSION/CONCLUSION: Adrenal incidentalomas patients with hypertension and normal hormone levels would perform hypertension resolution after laparoscopic adrenal tumor resection, especially for females with low BMI, high preoperative systolic blood pressure, and high APR (erect position).

Tislelizumab in combination with gemcitabine plus cisplatin chemotherapy as first-line adjuvant treatment for locally advanced or metastatic bladder cancer: a retrospective study.

BACKGROUND: Combining immune checkpoint inhibitors with chemotherapy can synergistically improve antitumor activity and are generally well tolerated. Recently, the efficacy and safety of combination therapy has been demonstrated for many cancers, including urothelial carcinomas. The aim of this retrospective pilot study was to evaluate the efficacy and safety of tislelizumab plus chemotherapy as first-line adjuvant treatment for locally advanced or metastatic bladder cancer. METHODS: We conducted a retrospective analysis of 31 patients with locally advanced or metastatic bladder cancer from December 2020 to January 2022 with an Eastern Cooperative Oncology Group performance status of 0/1. Of the 31 patients, 14 patients received tislelizumab (200 mg i.v. every 3 weeks, Q3W) plus 21 days cycles of chemotherapy (gemcitabine, 1000 mg/m2 i.v. on days 1 and 8 of each cycle + cisplatin, 70 mg/m2 i.v. on day 2 of each cycle) (TGC) treatment and 17 patients received gemcitabine plus cisplatin chemotherapy (GC) treatment. All patients treated with bladder cytoreductive surgery and were treated for four 21 days cycles until disease progression or intolerable treatment-related adverse events (TRAEs). The objective progression-free survival (PFS), overall survival (OS), overall response rate (ORR), disease control rate (DCR), clinical benefit rate (CBR) and TRAEs were recorded and reviewed. RESULTS: As of the cut-off date (March 25, 2022), PFS, OS, ORR, DCR, CBR and TRAEs were evaluated in 14 patients receiving combination therapy and 17 patients in the chemotherapy alone group. The median PFS was 36.0 [95% confidence interval (CI) 33.1-38.9] weeks in the TGC group and 29.0 (95% CI 25.4-32.6) weeks in the GC group [hazard ratio (HR) 0.15 (95% CI 0.04-0.55)]. In the GC group, the median OS was 48.0 (95% CI 39.7-56.3) weeks; the median OS was not yet mature for the TGC group [HR 0.26 (95% CI 0.07-0.94)]. Treatment with TGC resulted in improved DCR (TGC 71.4%; GC 65.0%) and CBR (TGC 64.3%; GC 52.9%) compared with GC. However, although higher incidences of grade ≥ 3 TRAEs were observed with TGC compared with GC (35.7% vs 23.5%), the difference was not statistically significant (p = 0.47). CONCLUSION: This study suggested that TGC provided survivors of locally advanced or metastatic bladder cancer with encouraging antitumor activity and was generally well tolerated.

P110ß Inhibition Reduces Histone H3K4 Di-Methylation in Prostate Cancer.

INTRODUCTION AND AIMS: Epigenetic alteration plays a major role in the development and progression of human cancers, including prostate cancer. Histones are the key factors in modulating gene accessibility to transcription factors and post-translational modification of the histone N-terminal tail including methylation is associated with either transcriptional activation (H3K4me2) or repression (H3K9me3). Furthermore, phosphoinositide 3-kinase (PI3 K) signaling and the androgen receptor (AR) are the key determinants in prostate cancer development and progression. We recently showed that prostate-targeted nano-micelles loaded with PI3 K/p110beta specific inhibitor TGX221 blocked prostate cancer growth in vitro and in vivo. Our objective of this study was to determine the role of PI3 K signaling in histone methylation in prostate cancer, with emphasis on histone H3K4 methylation. METHODS: PI3 K non-specific inhibitor LY294002 and p110beta-specific inhibitor TGX221 were used to block PI3 K/p110beta signaling. The global levels of H3K4 and H3K9 methylation in prostate cancer cells and tissue specimens were evaluated by Western blot assay and immunohistochemical staining. A synthetic androgen R1881 was used to stimulate AR activity in prostate cancer cells. A castration-resistant prostate cancer (CRPC) specific human tissue microarray (TMA) was used to assess the global levels of H3K4me2 methylation by immunostaining approach. RESULTS: Our data revealed that H3K4me2 levels were significantly elevated after androgen stimulation. With RNA silencing and pharmacology approaches, we further defined that inhibition of PI3 K/p110beta activity through gene-specific knocking down and small chemical inhibitor TGX221 abolished androgen-stimulated H3K4me2 methylation. Consistently, prostate cancer-targeted delivery of TGX221 in vivo dramatically reduced the global levels of H3K4me2 as assessed by immunohistochemical staining on tissue section of mouse xenografts from CRPC cell lines 22RV1 and C4-2. Finally, immunostaining data revealed a strong H3K4me2 immunosignal in CRPC tissues compared to primary tumors and benign prostate tissues. CONCLUSIONS: Taken together, our results suggest that PI3 K/p110beta-dependent signaling is involved in androgen-stimulated H3K4me2 methylation in prostate cancer, which might be used as a novel biomarker for disease prognosis and targeted therapy. Prostate 77:299-308, 2017. © 2016 Wiley Periodicals, Inc.

Clinical analysis of small cell carcinoma of the bladder in Chinese: nine case reports and literature reviews.

BACKGROUND: Small cell carcinoma of the bladder (SCCB) is a kind of rare and highly aggressive tumor that is present in an advanced stage and has a propensity for early metastasis. The main presenting symptom of SCCB is hematuria. Surgery, chemotherapy, and radiotherapy, either alone or as a part of combined therapy, have been used as the treatment. The aim of this study is to present our experience with 9 SCCB patients who were treated with different modalities and to share the findings upon reviewing the literatures for patients with SCCB reported in 56 literatures in Chinese. METHODS: We retrospectively evaluated 9 patients with SCCB from February 1980 to January 2014 in Tongji Hospital, Huazhong University of Science and Technology. The general characteristics, clinical manifestations, the pathological and immunohistochemical characteristics, treatment options, and prognostication in those eligible manuscripts were analyzed. In order to gain a better understanding of the clinical features of SCCB, another 119 cases reported in 56 articles were reviewed together (from January 1979 to March 2014). And a retrospective analysis was performed. RESULTS: All the 9 cases in Tongji Hospital were successfully operated, and the tissue samples were sent for pathological examination. All the tumor tissues contained small cell carcinoma components. 4 cases coexisted with other histologic types of bladder cancers, and 2 out of the 9 cases had three different cell components. All the patients had muscle invasion, and 4 cases showed lymph nodes metastasis, 3 cases showed invasion of neighboring structures (seminal vesicle or uterus), and 1 case was highly suspected of liver metastasis. Immunohistochemistry results showed that PCK, Syn, NSE, and CD56 were all positive, but LCA was negative. After operations, 3 patients underwent chemotherapy and only 1 patient received postoperative radiotherapy. Patients were followed up, ranging from 3 to 84 months and the median survival time was 33 months. The leading cause of death was tumor recurrence or metastasis, while 2 patients are still alive. According to the published literature, the pathological stage, immunohistochemical markers, and survival curves of all the 128 cases were also retrospectively analyzed. CONCLUSIONS: SCCB is different from transitional cell carcinoma (TCC) of the bladder. It has its unique cytology, immunohistochemistry, and ultrastructural features. Its diagnosis relies on pathological examination and immunohistochemistry. The current main treatment for SCCB is surgery combined with chemotherapy. Since the disease develops early metastasis easily, the overall prognosis of this cancer is poor. Further research need to clarify the molecular pathogenesis so that novel therapies can be developed for this rare cancer.

GSK-3ß controls autophagy by modulating LKB1-AMPK pathway in prostate cancer cells.

BACKGROUND: Glycogen synthase kinase 3ß (GSK3B, GSK-3ß) is a multi-functional protein kinase involved in various cellular processes and its activity elevates after serum deprivation. We have shown that inhibition of GSK-3ß activity triggered a profound autophagic response and subsequent necrotic cell death after serum deprivation in prostate cancer cells. In this study, we dissected the mechanisms involved in GSK-3ß inhibition-triggered autophagy. METHODS: Prostate cancer PC-3 and DU145 cells were used in the study. Multiple GSK-3ß specific inhibitors were used including small chemicals TDZD8, Tideglusib, TWS119, and peptide L803-mts. Western blot assay coupled with phospho-specific antibodies were used in detecting signal pathway activation. ATP levels were assessed with ATPLite kit and HPLC methods. Autophagy response was determined by evaluating Microtubule-associated proteins 1A/1B light chain 3B (LC3B) processing and p62 protein stability in Western blot assays. Immunofluorescent microscopy was used to detect LKB1 translocation. RESULTS: Inhibition of GSK-3ß activity resulted in a significant decline of cellular ATP production, leading to a significant increase of AMP/ATP ratio, a strong trigger of AMP-activated protein kinase (AMPK) activation in prostate cancer PC-3 cells. In parallel with increased LC-3B biosynthesis and p62 protein reduction, the classical sign of autophagy induction, AMPK was activated after inhibition of GSK-3ß activity. Further analysis revealed that Liver kinase B1 (LKB1) but not Calcium/calmodulin-dependent protein kinase kinase ß (CaMKKß) is involved in AMPK activation and autophagy induction triggered by GSK-3ß inhibition. Meanwhile, GSK-3ß inhibition promoted LKB1 translocation from nuclear to cytoplasmic compartment and enhanced LKB1 interaction with its regulatory partners Mouse protein-25 (MO25) and STE20-related adaptor (STRAD). CONCLUSIONS: In conclusion, our data suggest that GSK-3ß plays an important role in controlling autophagy induction by modulating the activation of LKB1-AMPK pathway after serum deprivation.

Preserved Erectile Function in the Aged Transgenic Rat Harboring Human Tissue Kallikrein 1.

INTRODUCTION: Human tissue kallikrein 1 (hKLK1) has enormous potential for the protection of vasodilation and endothelial function in the cardiovascular system. Our previous study proved the decreased expression of kallikrein 1 in the corpus cavernosum (CC) of aged rats, but the role of kallikrein 1 in age-related erectile dysfunction remains unknown. AIM: To explore the effect and underlying mechanisms of hKLK1 on age-related erectile dysfunction. METHODS: Male wild-type Sprague-Dawley rats (WTR) and transgenic rats harboring the hKLK1 gene (TGR) were fed to 4 and 27 months of age, respectively, and divided into four groups: young WTR (yWTR) as the control, young TGR (yTGR), aged WTR (aWTR), and aged TGR (aTGR). Rats' erectile function was evaluated by the cavernous nerve electrostimulation method. Then, CCs were collected for verification of hKLK1 followed by measurement of nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) and RhoA-Rho-kinase (ROCK) signaling activities. Masson trichrome staining and terminal deoxynucleotidyl transferase 2'-deoxyuridine 5'-triphosphate nick end labeling assay were conducted to evaluate penile fibrosis and apoptosis. MAIN OUTCOME MEASURES: Erectile response, NO-cGMP and RhoA-ROCK pathway-related indices, ratio of smooth muscle to collagen, and apoptosis index. RESULTS: The hKLK1 alleviated the decrease of erectile function in the aWTR group. Endothelial NO synthase (eNOS) and phospho-eNOS(Ser1177) expressions, NO synthase activity, and NO and cGMP levels were decreased, whereas phospho-eNOS(Thr495), L-type Ca(2+) channel, RhoA, ROCK1, ROCK2, and transforming growth factor ß1 proteins were increased in the CCs of the aWTR group compared with the control yWTR group. These changes were obviously mitigated in the aTGR group. Moreover, hKLK1 prevented the sharp decrease of the ratio of smooth muscle to collagen and the increase of the apoptosis index in the CCs of the aWTR group. CONCLUSION: These results suggest that hKLK1 could play a preventive role in age-related erectile dysfunction by activation of the NO-cGMP pathway and inhibition of the RhoA-ROCK pathway and by antitissue fibrotic and apoptotic effects.

Nanomicellar TGX221 blocks xenograft tumor growth of prostate cancer in nude mice.

BACKGROUND: Combination of androgen ablation along with early detection and surgery has made prostate cancer highly treatable at the initial stage. However, this cancer remains the second leading cause of cancer death among American men due to castration-resistant progression, suggesting that novel therapeutic agents are urgently needed for this life-threatening condition. Phosphatidylinositol 3-kinase p110ß is a major cellular signaling molecule and has been identified as a critical factor in prostate cancer progression. In a recent report, we established a nanomicelle-based strategy to deliver p110ß-specific inhibitor TGX221 to prostate cancer cells by conjugating the surface of nanomicelles with a RNA aptamer against prostate specific membrane antigen (PSMA) present in all clinical prostate cancers. In this study, we tested this nanomicellar TGX221 for its in vivo anti-tumor effect in mouse xenograft models. METHODS: Prostate cancer cell lines LAPC-4, LNCaP, C4-2 and 22RV1 were used to establish subcutaneous xenograft tumors in nude mice. Paraffin sections from xenograft tumor specimens were used in immunohistochemistry assays to detect AKT phosphorylation, cell proliferation marker Ki67 and proliferating cell nuclear antigen (PCNA), as well as 5-bromo-2-deoxyuridine (BrdU) incorporation. Quantitative PCR assay was conducted to determine prostate-specific antigen (PSA) gene expression in xenograft tumors. RESULTS: Although systemic delivery of unconjugated TGX221 significantly reduced xenograft tumor growth in nude mice compared to solvent control, the nanomicellar TGX221 conjugates completely blocked tumor growth of xenografts derived from multiple prostate cancer cell lines. Further analyses revealed that AKT phosphorylation and cell proliferation indexes were dramatically reduced in xenograft tumors received nanomicellar TGX221 compared to xenograft tumors received unconjugated TGX221 treatment. There was no noticeable side effect by gross observation or at microscopic level of organ tissue section. CONCLUSION: These data strongly suggest that prostate cancer cell-targeted nanomicellar TGX221 is an effective anti-cancer agent for prostate cancer.

Revealing the mechanism of ethyl acetate extracts of Semen Impatientis against prostate cancer based on network pharmacology and transcriptomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Prostate cancer (PCa) is the most common malignancy of the male genitourinary system and currently lacks effective treatment. Semen Impatientis, the dried ripe seed of Impatiens balsamina L., is described by the Chinese Pharmacopoeia as a traditional Chinese medicine (TCM) and is used in clinical practice to treat tumors, abdominal masses, etc. In our previous study, the ethyl acetate extracts of Semen Impatientis (EAESI) was demonstrated to be the most effective extract against PCa among various extracts. However, the biological effects of EAESI against PCa in vivo and the specific antitumor mechanisms involved remain unknown. AIM OF THE STUDY: In this study, we aimed to investigate the antitumor effect of EAESI on PCa in vitro and in vivo by performing network pharmacology analysis, transcriptomic analysis, and experiments to explore and verify the underlying mechanisms involved. MATERIALS AND METHODS: The antitumor effect of EAESI on PCa in vitro and in vivo was investigated via CCK-8, EdU, flow cytometry, and wound healing assays and xenograft tumor models. Network pharmacology analysis and transcriptomic analysis were employed to explore the underlying mechanism of EAESI against PCa. Activating transcription factor 3 (ATF3) and androgen receptor (AR) were confirmed to be the targets of EAESI against PCa by RT‒qPCR, western blotting, and rescue assays. In addition, the interaction between ATF3 and AR was assessed by coimmunoprecipitation, immunofluorescence, and nuclear-cytoplasmic separation assays. RESULTS: EAESI decreased cell viability, inhibited cell proliferation and migration, and induced apoptosis in AR+ and AR- PCa cells. Moreover, EAESI suppressed the growth of xenograft tumors in vivo. Network pharmacology analysis revealed that the hub targets of EAESI against PCa included AR, AKT1, TP53, and CCND1. Transcriptomic analysis indicated that activating transcription factor 3 (ATF3) was the most likely critical target of EAESI. EAESI downregulated AR expression and decreased the transcriptional activity of AR through ATF3 in AR+ PCa cells; and EAESI promoted the expression of ATF3 and exerted its antitumor effect via ATF3 in AR+ and AR- PCa cells. CONCLUSIONS: EAESI exerts good antitumor effects on PCa both in vitro and in vivo, and ATF3 and AR are the critical targets through which EAESI exerts antitumor effects on AR+ and AR- PCa cells.

Up-regulation of VEGF by small activator RNA in human corpus cavernosum smooth muscle cells.

INTRODUCTION: Functional failure of smooth muscle cells and endothelial cells in corpus cavernosum contributes to erectile dysfunction (ED) in aging men. Given that vascular endothelial growth factor (VEGF) may improve the function of smooth muscle cells and endothelial cells through different mechanisms, it is thus expected that increasing the expression of VEGF may have beneficial effects on erectile function. AIM: The aim of this article is to explore the possibility that VEGF can be induced by ribonucleic acid activation (RNAa) technology, and VEGF induction by RNAa has the potential of treating ED. METHODS: Primary human corpus cavernosum smooth muscle cells (CCSMCs) were isolated and cultured in vitro. The expression of α-smooth muscle actin was detected by immunohistochemistry to identify CCSMCs. A previously identified VEGF promoter-targeted small activator RNA (saRNA, double-stranded [ds]VEGF-706) and a negative control dsRNA were chemically synthesized. Cultured human CCSMCs were transfected with the saRNAs. The expression of VEGF messenger RNA (mRNA) and protein in transfected CCSMCs was evaluated by real-time polymerase chain reaction (RT-PCR) and Western blotting assay, respectively. Immunofluorescent staining was also used to confirm VEGF protein expression in cultured CCSMCs. MAIN OUTCOME MEASURE: The expression of VEGF was assessed by RT quantitative PCR, Western blotting, and immunofluorescence assays. RESULTS: After transfection, RT quantitative PCR analysis showed that the expression of VEGF mRNA was significantly induced in dsVEGF-706 transfected cells compared with cells receiving control treatments (P < 0.05). Consistent with mRNA induction, Western blotting and immunofluorescence analysis showed that VEGF protein expression was also induced by dsVEGF-706. CONCLUSION: VEGF expression can be activated by RNAa in primary human CCSMCs, suggesting a potential application of RNAa-mediated VEGF activation for the treatment of ED.

Optimize the management of urological tube-related emergencies during the coronavirus disease 2019 (COVID-19) pandemic.

BACKGROUND: To introduce and determine the value of optimized strategies for the management of urological tube-related emergencies with increased incidence, complexity and operational risk during the global spread of coronavirus disease 2019 (COVID-19). METHODS: All emergent urological patients at Tongji Hospital, Wuhan, during the period of January 23 (the beginning of lockdown in Wuhan) to March 23, 2020, and the corresponding period in 2019 were recruited to form this study's COVID-19 group and control group, respectively. Tongji Hospital has the most concentrated and strongest Chinese medical teams to treat the largest number of severe COVID-19 patients. Patients in the control group were routinely treated, while patients in the COVID-19 group were managed following the optimized principles and strategies. The case incidence for each type of tube-related emergency was recorded. Baseline characteristics and management outcomes (surgery time, secondary complex operation rate, readmission rate, COVID-19 infection rate) were analyzed and compared across the control and COVID-19 periods. RESULTS: The total emergent urological patients during the COVID-19 period was 42, whereas during the control period, it was 124. The incidence of tube-related emergencies increased from 53% to 88% (P<0.001) during the COVID-19 period. In particular, the incidence of nephrostomy tube-related (31% vs. 15%, P=0.027) and single-J stent-related problems (19% vs. 6%, P=0.009) increased significantly. The mean surgery times across the two periods were comparable. The number of secondary complex operations increased from 12 (18%) to 14 (38%) (P=0.028) during the COVID 19-period. The number of 2-week postoperative readmission decreased from 10 (15%) to 1 (3%) (P=0.049). No participants contracted during the COVID-19 period. CONCLUSIONS: Urological tube-related emergencies have been found to have a higher incidence and require more complicated and dangerous operations during the COVID-19 pandemic. However, the optimized management strategies introduced in this study are efficient, and safe for both urologists and patients.

Solitary fibrous tumor in bladder: a case report.

Solitary fibrous tumor (SFT) in bladder is extremely rare. In this study, we reported one case of bladder SFT and reviewed the only ten cases of the disease that had been reported so far. The patient suffered from residual urine sensation and urethral pain. Cystoscopy revealed a 7-cm protruding mass at the dome of the bladder, and bladder mucosa biopsy showed normal differentiation of the bladder mucosa with a small amount of inflammatory cells. Radical resection of the tumor was performed in this patient. Pathological examination found uniform, haphazardly arranged spindle cells, the majority of which were CD34-positive and Vimentin-positive and proved that the mass was a solitary fibrous tumor. Within a period of 9 months of follow-up, no reoccurrence was found.

[Changes of contractility and relaxation of corpus cavernosum in hyperlipidemic rabbit in vitro].

OBJECTIVE: To investigate the mechanisms of erectile dysfunction induced by hyperlipidemia through studying the changes of contractility and relaxation of corpus cavernosum in hypercholesterolemic rabbit in vitro. METHODS: New Zealand white rabbits were randomly divided into control and experiment groups. The control group (n = 20) received a regular diet for 8 weeks while the experimental group (n = 20) were fed a 1% cholesterol diet for 8 weeks. The authors conducted isometric tension studies with phenylephrine, endothelium-dependent vasodilators (acetylcholine), endothelium-independent vasodilators (sodium nitroprusside) and rho kinase inhibitor (Fasudil) on isolated strips of corpus cavernosum. RESULTS: The weight and total serum cholesterol significantly improved (P < 0.01) in experimental group as compared with the non-fed experimental group. The total serum cholesterol significantly improved (P < 0.01) in experimental group as compared with the control group. The contractility responses to phenylephrine in experiment group in doses (0.5, 1, 5, 10, 50, 100 micromol) were 4.79% +/- 2.00%, 8.84% +/-2.95%, 12.81% +/- 3.77%, 14.63% +/- 5.38%, 25.01% +/- 6.14% and 34.69% +/- 8.53% respectively. The contractility responses to phenylephrine in control group were 1.00% +/- 0.3%, 2.60% +/- 0.72%, 4.28% +/- 1.27%, 5.91% +/- 2.09%, 6.49% +/- 4.02% and 5.64% +/- 11.87% respectively. The contractility responses to phenylephrine significantly improved (P < 0.01) in experimental group in these doses as compared with the control group. The relaxation responses to acetylcholine in experimental group in doses (1, 10 and 100 micromol) were 36.28% +/- 4.71%, 48.81% +/- 4.36% and 56.27% +/- 11.93% respectively. The relaxation responses in control group were 48.04% +/- 4.78%, 69.12% +/- 5.27% and 78.23% +/- 5.30% respectively. There was significant reduction (P < 0.01) in experimental group in these doses as compared with the control group. No difference were found among the two groups in the relaxation response to sodium nitroprusside. The relaxation responses to fasudil in experimental group in doses (0.25, 1.25, 2.5 and 12.5 micromol) were 1.56% +/- 0.43%, 5.03% +/- 1.02%, 8.28% +/- 1.35% and 16.77% +/- 3.57% respectively. The relaxation responses in control group were 4.69% +/- 1.23%, 10.39% +/- 2.05%, 15.08% +/- 3.04% and 25.22% +/- 3.72% respectively. There was significant reduction (P < 0.01) in experimental group in these doses as compared with the control group. CONCLUSION: The improvement of cavernous smooth muscle contractility and the impairment of cavernous smooth muscle relaxation in response to endothelium-mediated stimuli are the mechanisms of erectile dysfunction induced by hyperlipidemia.

[Differentiation of rat adipose-derived stem cells into smooth-muscle-like cells in vitro].

OBJECTIVE: To study the feasibility of inducing the differentiation of rat adipose-derived stem cells (ADSCs) into smooth-muscle-like cells in monolayer culture in vitro. METHODS: ADSCs were obtained from the adipose of the inguinal region of the SD rat. A growth curve of the ADSCs was drawn. The fourth passage ADSCs were induced to differentiate into adipocytes with adipogenic inducing fluid and determined by Oil Red O staining, into osteoblasts with osteogenic inducing fluid and determined by Von Kossa staining, as well as into smooth-muscle-like cells with inducing fluid containing beta-mercaptoethanol, and the expression of alpha-smooth muscle actin (alpha-SMA) was detected by the immunohistochemical method. RESULTS: The ADSCs presented spindle and polygon shapes and proliferated rapidly in vitro. The growth curve showed the ADSCs in the logarithmic growth phase two days after subculture. The fourth passage ADSCs exhibited red stained lipid droplets characteristic of adipocytes in the cytoplasm on Oil Red O staining after induction with adipogenic inducing fluid, and calcium nodes on Von Kossa staining after induction with osteogenic inducing fluid. The immunohistochemical results of the ADSCs induced into smooth-muscle-like cells showed that the positive rate of alpha-SMA in the beta-mercaptoethanol induced cells was (29.80 +/- 6.89)%, significantly higher than in the uninduced ones ([2.89 +/- 1.24]%, P < 0.01). CONCLUSION: ADSCs displayed obvious characteristics of smooth muscle cells after induction, and could be a new source of cells in the tissue engineering studies of smooth muscle related diseases.

Methylene Blue Injection via Percutaneous Renal Cyst Puncture Used in Flexible Ureteroscope for Treatment of Parapelvic Cysts: A Modified Method for Easily Locating Cystic Wall.

OBJECTIVE: To describe a modified technique for easily locating cystic wall during flexible ureteroscopic surgery for treatment of parapelvic renal cysts. METHODS: Nineteen patients with symptomatic/asymptomatic parapelvic renal cyst were treated with modified or conventional flexible ureteroscopic surgery between February 2015 and March 2017, and the differences of the 2 techniques were compared. The detailed surgical procedures and results, postoperative complications, and patients' follow-ups were evaluated. RESULTS: All the patients received endoscopic management by flexible ureteroscope successfully, without requiring another complicated surgery. The cysts were seen clearly in 9 patients with modified method. Two of ten patients who underwent conventional ureteroscopic surgery changed to the modified surgery intra-operatively since it was difficult to identify the cyst. The total time of search and incision of cysts was 24.2 ± 6.2 minutes and 17.7 ± 2.5 minutes for conventional and modified technique respectively (P = .01), of which 11.4 ± 4.8 minutes and 5.1 ± 1.1 minutes to search the cysts, respectively (P = .002), and the mean time of the procedure of puncture was 8 ± 2.3 minutes. Duration of pure incising the cystic wall was 12.8 ± 3.3 minutes and 12.6 ± 2.5 minutes for patients who underwent conventional and modified technique, respectively (P = .859). All patients acquired relief from the presentation of flank discomfort after surgery. All of the patients were followed-up over 12 months and no serious complications and recurrence was observed. CONCLUSION: The modified technique can decrease time of searching the renal cyst and decrease the total time in flexible ureteroscopic treatment of parapelvic cysts. The limitations of our study were also observed and further studies are needed.

Xanthine oxidase-mediated oxidative stress promotes cancer cell-specific apoptosis.

The natural compound Alternol was shown to induce profound oxidative stress and apoptotic cell death preferentially in cancer cells. In this study, a comprehensive investigation was conducted to understand the mechanism for Alternol-induced ROS accumulation responsible for apoptotic cell death. Our data revealed that Alternol treatment moderately increased mitochondrial superoxide formation rate, but it was significantly lower than the total ROS positive cell population. Pre-treatment with mitochondria-specific anti-oxidant MitoQ, NOX or NOS specific inhibitors had no protective effect on Alternol-induced ROS accumulation and cell death. However, XDH/XO inhibition by specific small chemical inhibitors or gene silencing reduced total ROS levels and protected cells from apoptosis induced by Alternol. Further analysis revealed that Alternol treatment significantly enhanced XDH oxidative activity and induced a strong protein oxidation-related damage in malignant but not benign cells. Interestingly, benign cells exerted a strong spike in anti-oxidant SOD and catalase activities compared to malignant cells after Alternol treatment. Cell-based protein-ligand engagement and in-silicon docking analysis showed that Alternol interacts with XDH protein on the catalytic domain with two amino acid residues away from its substrate binding sites. Taken together, our data demonstrate that Alternol treatment enhances XDH oxidative activity, leading to ROS-dependent apoptotic cell death.

Ki-67 index and percentage of sarcomatoid differentiation were two independent prognostic predictors in sarcomatoid renal cell carcinoma.

PURPOSE: To identify whether and which of pathological features of sarcomatoid differentiation (SD) in renal cell carcinoma (RCC) can be used as independent predictors associated with overall survival (OS). MATERIALS AND METHODS: After institutional review board approval, patients with a diagnosis of sarcomatoid RCC (sRCC), spindled RCC, or RCC with the presence of spindle cells between 2003 and 2017 were further selected and re-examined. The primary pathological features including histological subtypes, tumor necrosis, Ki-67 index of SD, and the percent of SD (%SD) were included into analysis. Histological subtypes were categorized into clear-cell RCC and nonclear-cell RCC. Ki-67 index of SD was confirmed by immunohistochemical staining. %SD was estimated through reviewing all of the tumor sections microscopically and then giving an approximate %SD within the entire tumor. The clinical relevant prognostic predictor's association with OS was analyzed within Cox proportional hazards regression models. Survival curves were generated using the Kaplan-Meier method, and OS differences were compared using the log-rank test. RESULTS: A total of 2,089 consecutive patients of RCC were referred to our department, of whom 62 (3.0%) patients were identified with histological element of SD after re-examining the available slides of suspicious cases. Finally, 53 patients were included into survival analysis after excluding 9 patients without adequate information. Thirty-eight (71.7%) patients died at last follow-up. The median OS for all patients was 11.0 months from the date of surgery. In patients with clinical distant metastasis (cM1), the median OS was only 3 compared with 21 months for patients with no clinical distant metastasis (cM0). Tumor stage, status of clinical distant metastasis, Ki-67 index, and %SD were independent predictors of multivariate analysis in overall 53 patients. However, in the cohort of cM0 patients, we found that only %SD and Ki-67 index were two independent predictors of OS in multivariate analysis. CONCLUSION: Patients with sRCC are associated with very poor prognosis. Ki-67 index of SD and %SD were identified as the two most important independent predictors particularly for nonmetastatic patients. The limitations of our study were also observed, and further studies are needed.

Will the kidney function be reduced in patients with renal cell carcinoma following laparoscopic partial nephrectomy? Baseline eGFR, warm ischemia time, and RENAL nephrometry score could tell.

OBJECTIVES: To describe the natural history of kidney function following partial nephrectomy (PN) for patients with renal cell carcinoma (RCC), and to identify independent predictors of whether patients with RCC will retain renal function unchangeable or even increased and develop functional impairment of ≧25% post-PN. PATIENTS AND METHODS: We performed a retrospective analysis of 337 cases involving patients diagnosed with RCC of pT1-2N0M0 who underwent laparoscopic PN, the primary endpoints included the stabilization or increase in postoperative estimated glomerular filtration rate (eGFR) compared to the preoperative level and eGFR impairment of ≧25% following surgery. We plotted the trajectory of each patient's eGFR measurement starting from their first postoperative day to the last follow-up time post-PN and used moving average method to look at trends of eGFR changing. A logistic regression model was then applied to identify associations between clinical and surgical characteristics with eGFR outcomes. RESULTS: Patients were of an average age of 51.4 years and all were Chinese descent. The cohort was also primarily male (69.1%). One hundred ninety seven (58.5%) had eGFR ≧90 ml/min/1.73 m2, while 140 (41.5%) had an eGFR of 60 to 90 ml/min/1.73 m2 prior to the operation. All patients underwent minimally invasive PN with warm ischemia, with 64.1% (216/337) receiving laparoscopic surgery, and 35.9% (121/337) receiving robot-assisted laparoscopic surgery. On average, patients experienced a mean eGFR decrease of 23.8% immediately post-PN, followed by a slight increase and stabilization, with a mean 15.5% decline after 1 year. Twenty four percent (81/337) experienced GFR impairment of ≧25% over a median 10.0-month follow-up time period, while 29.1% (99/337) patients retained eGFR unchangeable or increased post-PN. And higher preoperative eGFR, longer warm ischemia time, and more complexity lesions (higher renal nephrometry score ) were found to be independently associated with higher chance of functional impairment of ≧25% and lower chance of eGFR stabilization post-PN. CONCLUSION: Although, majority of patients experienced decline of renal function post-PN, functional outcomes of eGFR unchangeable and increased were also seen, and baseline total eGFR, WIT as well as RENAL nephrometry score were determined to be independent predictors of those renal functional outcomes.

Free bladder mucosa graft harvested by water-jet: A novel, minimally invasive technique for urethral reconstruction.

The aim of the current study was to describe a novel approach of urethral reconstruction through minimally invasive harvesting of the bladder graft via endoscopic sub-mucosal dissection of water-jet. The records of two patients were reviewed, who underwent transurethral endoscopic surgical bladder mucosa graft harvest by water-jet and urethral reconstruction with informed consent. Case 1 was a 35-year-old male with anterior urethral stricture; case 2 was a 22-year-old male with secondary anterior urethral stricture and hypospadias following a failed hypospadias surgery. The two male patients successfully underwent urethral reconstruction using bladder mucosa graft harvested via endoscopic assisted by water-jet; no perforation, cysthemorrhagia or any other postoperative bladder-related complication was observed. Voiding cystourethrogram of case 1 indicated that the reconstructed urethra was unobstructed, and no recrudescence was observed within 4 months of follow-up. In case 2, dysuria had disappeared completely within 1 month of follow-up, and the urethra plate was successfully reconstructed by first-stage. To the best of our knowledge, this is the first report to demonstrate urethral reconstruction using a bladder mucosa graft harvested by transurethral endoscopic sub-mucosal dissection, assisted by water-jet. Transurethral endoscopic surgery may provide a minimally invasive approach instead of the traditional open surgery for harvesting bladder mucosa graft. Urethral reconstruction conducted with bladder mucosa graft harvested via endoscopic sub-mucosal dissection assisted by water-jet is a feasible and safe method, and the short-term follow-up results are encouraging.

Predictive value of FSH, testicular volume, and histopathological findings for the sperm retrieval rate of microdissection TESE in nonobstructive azoospermia: a meta-analysis.

We performed this meta-analysis to evaluate the predictive value of different parameters in the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (TESE) in patients with nonobstructive azoospermia (NOA). All relevant studies were searched in PubMed, Web of Science, EMBASE, Cochrane Library, and EBSCO. We chose three parameters to perform the meta-analysis: follicle-stimulating hormone (FSH), testicular volume, and testicular histopathological findings which included three patterns: hypospermatogenesis (HS), maturation arrest (MA), and Sertoli-cell-only syndrome (SCOS). If there was a threshold effect, only the area under the summary receiver operating characteristic curve (AUSROC) was calculated. Otherwise, the pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and the diagnostic odds ratio (DOR) were also calculated. Twenty-one articles were included in our study finally. There was a threshold effect among studies investigating FSH and SCOS. The AUSROCs of FSH, testicular volume, HS, MA, and SCOS were 0.6119, 0.6389, 0.6758, 0.5535, and 0.2763, respectively. The DORs of testicular volume, HS, and MA were 1.98, 16.49, and 1.26, respectively. The sensitivities of them were 0.80, 0.30, and 0.27, while the specificities of them were 0.35, 0.98, and 0.76, respectively. The PLRs of them were 1.49, 10.63, and 1.15, respectively. And NLRs were 0.73, 0.72, and 0.95, respectively. All the investigated factors in our study had limited predictive value. However, the histopathological findings were helpful to some extent. Most patients with HS could get sperm by microdissection TESE.

Modulating circulating sFlt1 in an animal model of preeclampsia using PAMAM nanoparticles for siRNA delivery.

INTRODUCTION: Excessive circulating sFlt1 plays a major role in the pathogenesis of preeclampsia (PE). Using RNAi to silence sFlt1 may be a therapy for treating PE. Because of the rapid degradation of siRNA, gene therapy in vivo remains limited. Poly-amidoamine (PAMAM) has been demonstrated to be an excellent nanocarrier for siRNA delivery with no discernible toxicity. METHODS: The aim of the present study was to investigate the therapeutic effect of siRNA-sFlt1-PAMAM on PE. The biophysical properties of siRNA-sFlt1-PAMAM complexes were analysed. Next, HTR-8/SVneo cells were incubated with the complexes. The transfection efficiency, silencing effect, and cell proliferation were examined. The timed pregnant rats were grouped to test the in vivo effect of siRNA-sFlt1-PAMAM on PE. The PE model was established by injection of tumour necrosis factor-α (TNF-α), while treatment group rats were injected with both TNF-α and siRNA-sFlt1-PAMAM. The mean arterial pressure, serum levels of sFlt1, and 24-h urinary protein were measured. The number of viable pups, and weights of individual pups and placentae were determined. RESULTS: siRNA-sFlt1-PAMAM complexes showed high cellular uptake and excellent siRNA encapsulation ability. siRNA-sFlt1-PAMAM significantly decreased sFlt1 secretion from HTR-8/SVneo cells (p = 0.001) with little effect on HTR-8/SVneo cell proliferation. The circulating level of sFlt1, mean arterial pressure, and urine protein level in the TNF-α group were significantly increased compared to that in the control group, while the weights of foetuses and placentae were significantly decreased compared to that in the control group (p < 0.05). All preeclamptic symptoms were greatly attenuated by siRNA-sFlt1-PAMAM (p < 0.05). CONCLUSIONS: These data suggest that siRNA-sFlt1-PAMAM effectively decreased sFlt1 secretion and improved pregnancy outcomes in a preeclamptic rat model, which may provide a new therapeutic strategy for PE.