ORC6, a novel prognostic biomarker, correlates with T regulatory cell infiltration in prostate adenocarcinoma: a pan-cancer analysis.

BACKGROUND: The origin recognition complex (ORC), a six-subunit DNA-binding complex, participates in DNA replication in cancer cells. Specifically in prostate cancers, ORC participates the androgen receptor (AR) regulated genomic amplification and tumor proliferation throughout the entire cell cycle. Of note, ORC6, the smallest subunit of ORC, has been reported to be dysregulated in some types of cancers (including prostate cancer), however, its prognostic and immunological significances remain yet to be elucidated. METHODS: In the current study, we comprehensively investigated the potential prognostic and immunological role of ORC6 in 33 human tumors using multiple databases, such as TCGA, Genotype-Tissue Expression, CCLE, UCSC Xena, cBioPortal, Human Protein Atlas, GeneCards, STRING, MSigDB, TISIDB, and TIMER2 databases. RESULTS: ORC6 expression was significantly upregulated in 29 types of cancers compared to the corresponding normal adjacent tissues. ORC6 overexpression correlated with higher stage and worse prognostic outcomes in most cancer types analyzed. Additionally, ORC6 was involved in the cell cycle pathway, DNA replication, and mismatch repair pathways in most tumor types. A negative correlation was observed between the tumor endothelial cell infiltration and ORC6 expression in almost all tumors, whereas the immune infiltration of T regulatory cell was noted to be statistically positively correlated with the expression of ORC6 in prostate cancer tissues. Furthermore, in most tumor types, immunosuppression-related genes, especially TGFBR1 and PD-L1 (CD274), exhibited a specific correlation with the expression of ORC6. CONCLUSIONS: This comprehensive pan-cancer analysis revealed that ORC6 expression serves as a prognostic biomarker and that ORC6 is involved in the regulation of various biological pathways, the tumor microenvironment, and the immunosuppression status in several human cancers, suggesting its potential diagnostic, prognostic, and therapeutic value in pan-cancer, especially in prostate adenocarcinoma.

Application of a new position in endoscopic combined intrarenal surgery: modified prone split-leg position.

BACKGROUND: Endoscopic combined intrarenal surgery (ECIRS) is well established as a minimally invasive procedure for the treatment of multiple urolithiasis. The position is the key to the perfect combination of percutaneous nephrolithotomy (PCNL) and retrograde intrarenal surgery (RIRS). Galdakao-modified supine Valdivia (GMSV) and prone split-leg positions are widely used. However, both positions have their own advantages and disadvantages. This study aimed to evaluate the effect of ECIRS in the treatment of multiple urolithiasis in the modified prone split-leg position. PATIENTS AND METHODS: A total of 96 patients with multiple urolithiasis underwent ECIRS in modified prone split-leg position from September 2017 to January 2021. Relevant demographic and clinical data were analysed retrospectively. Clinical outcomes, such as the stone free rate, complications and postoperative hospital stay were evaluated. The chi-square test was used to compare categorical variables and Student's t test was applied for continuous variables of the treatment groups. RESULTS: The mean renal stone size was 32.5 ± 10.7 mm and renal stone surface area was 712.2 ± 264.8 mm2. The mean ureteral stones size was 24.8 ± 12.3 mm. The mean surgical time was 82.2 ± 38.3 min. The incidence of complications was 16.7%, and they were mainly grade 1 and grade 2. No complications occurred above grade 3. The stone was completely removed in 75 (78.1%) patients in a single operation. The risk factors affecting the stone-free rate of ECIRS were analysed, and only the number of involved calyces by stone was found to be significant (p = 0.01). CONCLUSION: ECIRS is safe and effective in the treatment of multiple renal calculi or multiple renal calculi with ipsilateral ureteral calculi in the modified prone split-leg position. The modification of the prone split-leg position makes the retrograde operation more convenient, which is conducive to the combination of RIRS and PCNL.

Endoscopic combined intrarenal surgery in the prone-split leg position for successful single session removal of an encrusted ureteral stent: a case report.

BACKGROUND: The most serious complication of ureter stent is long-term retention of ureteral stent and stone formation around the stent. CASE PRESENTATION: A 51-year old female patient with left ureteral stent placed 2 years before developed both pyelic and vesical stones on the two ends of the double J was admitted to our hospital. Intravesical lithotripsy, retrograde ureteroscopy, and percutaneous nephrolithotripsy were performed with the patient in the prone split-leg position. All the stones and the ureteral stent were successfully removed in a single session. CONCLUSIONS: Combined endoscopic techniques in the prone split-leg position can effectively and safely manage severely encrusted stents.

Melatonin alleviates intrarenal CaOx crystals deposition through inhibiting LPS-induced non-canonical inflammasome-mediated renal tubular epithelial cells pyroptosis.

Urinary tract infection has long been considered a complication rather than etiology of calcium oxalate (CaOx) nephrolithiasis. This study aimed to explore the role of lipopolysaccharide (LPS), an important component of Gram-negative bacteria, on CaOx nephrolithiasis formation and antagonistic effect of melatonin. Male C57BL/6 mice were intraperitoneally injected with glyoxylate acid (80 mg/kg) daily for 7 days to construct CaOx nephrolithiasis model. A single dose of LPS (2.0 mg/kg) was given 2 h before the second glyoxylate acid treatment in the presence or absence of melatonin (25 mg/kg). Our results found that LPS promoted adhesion of CaOx crystals to renal tubular epithelial cells (RTECs) and intrarenal CaOx crystals deposition. Protein levels of cleaved Caspase-11, N-terminal of cleaved GSDMD (GSDMD-N), NOD-like receptor thermal protein domain associated protein 3 (NLRP3) and cleaved Caspase-1, several markers of non-classical inflammasome activation were upregulated in LPS-treated mouse kidneys and HK-2 cells. Moreover, the number of GSDMD pores was increased in LPS-treated HK-2 cell membrane. Melatonin inhibited Caspase-11 cleavage and antagonized the subsequent LPS-mediated upregulation of GSDMD-N, NLRP3 and cleaved Caspase-1 in kidney tissues and HK-2 cells. In addition, melatonin reduced membrane localization of GSDMD-N and the number of GSDMD pores in LPS-treated HK-2 cells. Accordingly, melatonin inhibited LPS-induced IL-1ß and IL-18 in mouse serum and HK-2 culture supernatant. Importantly, melatonin alleviated LPS-induced crystal-cell interactions and intrarenal CaOx crystals deposition. We provide experimental evidence that LPS promoted CaOx nephrolithiasis formation by inducing non-canonical inflammasome-mediated RTECs pyroptosis. Melatonin alleviated CaOx nephrolithiasis formation through inhibiting LPS-induced non-canonical inflammasome-mediated RTECs pyroptosis.

The Clinical Efficacy and Safety of Extracorporeal Shock Wave Lithotripsy in the Treatment of Patients with Urinary Calculi.

Objective: To evaluate the clinical efficacy of extracorporeal shock wave lithotripsy (ESWL) for urinary calculi and precautions of postoperative complications. Methods: 90 patients with urinary calculi at our hospital were randomly recruited between July 2019 and July 2020 and were allocated (1 : 1) to receive either ESWL (observation group) or conventional surgery (control group). Clinical efficacy was the primary endpoint, whereas adverse events were the secondary endpoint. Results: The operation time, early activity time, and hospitalization time of the observation group were significantly lower than those of the control group (P < 0.05). ESWL resulted in less postoperative pain in patients versus conventional surgery (P < 0.05). ESWL was associated with a significantly higher total clinical efficacy (97.78%) versus conventional surgery (82.22%) (P < 0.05). The eligible patients given ESWL had a lower incidence of complications (11.12%) versus those given conventional surgery (31.12%) (P < 0.05). Conclusion: Hematuria prevention requires precise localization of stones as well as adjustment of pulse energy and the number of impacts due to stone changes. Precautions against renal colic necessitate complete comminution of stones intraoperatively, more postoperative water intake, moderate exercise, or injection of antispasmodic drugs and cathartics for pain relief. Nausea and vomiting precautions require preoperative recording of previous medical history and corresponding treatment, intraoperative real-time adjustment of voltage pulse frequency, and duration depending on the magnitude of intraoperative reaction. Urinary tract infection prevention requires preoperative prevention and proper postoperative anti-infection and anti-inflammatory treatment, along with enough water intake and bed rest. Other precautions include thorough comminution of the calculi, proper anti-infection and anti-inflammatory treatment, no early exercise or excessive activity after surgery, and proper postoperative care. ESWL is effective in treating patients with urinary calculi with a simple, safe, and quick operation and a low incidence of adverse events, as it effectively reduces the incidence of complications, accelerates the recovery of patients and improves their quality of life.

p-Phenylenediamine induces epithelial-mesenchymal transition in SV-40 immortalized human urothelial cells via the ERK5/AP-1 signaling.

PURPOSE: To investigate whether p-Phenylenediamine (PPD) could triggered EMT inSV-40 immortalized human urothelial cells (SV-HUC-1), and the regulation role of ERK5/AP-1 during this process. MATERIALS AND METHODS: SV-HUC-1 cells were treated with different concentrations of PPD. MTT assay was employed to detect cell viability. Wound healing and transwell assay were performed to detect migrative and invasive capacity. Western blot and qRT-PCR were utilized for detecting molecular changes. ERK5 specific inhibitor was used to suppress ERK5 signaling. RESULTS: Migration and invasion capacity of SV-HUC-1cells were enhanced after PPD exposure. Expression of epithelial markers E-cadherin and ZO-1 was decreased and expression of mesenchymal markers N-cadherin and vimentin was increased after being cultured with low concentrations of PPD, indicating that PPD induced EMT in PPD-cultured SV-HUC-1 cells. Meanwhile, PPD triggered activation of ERK5signaling and downstream AP-1 was activated, but no obvious influence of PPD on other sub-families of MAPK was detected. After inhibition of ERK5/AP-1, PPD-induced enhancement of migrative and invasive abilities were attenuated and expression of EMT markers was also reversed. CONCLUSION: PPD may be a carcinogen, which could induce EMT in SV-40 immortalized human urothelial cells (SV-HUC-1) via activating ERK5/AP-1 signaling.

Sirt1 inhibits kidney stones formation by attenuating calcium oxalate-induced cell injury.

Cell injury is a necessary and critical event during CaOx kidney stone formation. Sirt1 exerts a number of pleiotropic effects, protecting against renal cell injury. This study aims to explore the relationship between Sirt1 and CaOx kidney stone formation and the underlying mechanism. Sirt1 expression in renal tissues or HK-2 cells was detected by Western blot, immunohistochemistry and immunofluorescence. Apoptosis in renal tissues was examined by TUNEL staining. Renal pathological changes and the crystals deposition were detected by hematoxylin-eosin and Von Kossa staining. Crystal-cell adhesion and cell injury in HK-2 cells were assessed by atomic absorption spectrometry and flow cytometry, respectively. Sirt1 expression in nephrolithiasis patients was downregulated and the level of apoptosis was increased. Further study found that Sirt1 expression was decreased in both in vivo and in vitro models. Interestingly, the levels of cell injury were elevated in vivo and in vitro models. Suppressing Sirt1 expression promoted COM-induced crystal-cell adhesion and exacerbated cell injury. In contrast, increasing the expression of Sirt1 by lentivirus transfection in vitro and resveratrol administration in vivo, alleviated crystal deposition and cell damage. Our findings suggest that Sirt1 could inhibit kidney stone formation, at least in part, through attenuating CaOx -induced cell injury.

Role of PI3K/Akt pathway in Benzidine-induced proliferation in SV-40 immortalized human uroepithelial cell.

BACKGROUND: Long term exposure to benzidine has been determined as a cause of urothelial carcinoma. But how it works in the process of cell proliferation that involves in tumor growth is not examined yet. In the current research, the effect of PI3K/Akt on cell proliferation mediated by benzidine was confirmed. METHODS: The immortalized SV-40 human uroepithelial cells (SV-HUC-1) had been subjected to 6 days of benzidine treatment at various contents, then MTT assay, together with subsequent flow cytometry assay were used for observing effects on cell proliferation. Further Western blots were used to detect the expression of total-Akt, phospho-Akt and specific proteins of cell cycle. The Akt, Cyclin D1, PCNA and P21 mRNA levels were detected through RT-PCR. In addition, the blocker-LY294002 was used to cut down the PI3K/Akt signaling pathway. And then those parameters were detected using the same methods as above. RESULTS: Results showed that benzidine acted to induce cell proliferation at low doses (P<0.05 vs. controls) via MTT and flow cytometry assay. The expression of phospho-Akt, Cyclin D1, and PCNA were significantly enhanced compared with that of control (P<0.05; P<0.01), but total-Akt and P21 levels were reduced. Whereas, inhibitor of PI3K/Akt suppressed the proliferating procedure when cells were treated with the blocker (LY294002) and also inhibited the expression of related cycle proteins. CONCLUSIONS: Activated PI3K/Akt signal pathway promotes benzidine-triggered cell proliferation. It may shed light on the molecular mechanisms that the activated PI3K/Akt pathway promotes benzidine-triggered cell proliferation and intervention of its target.

Resveratrol Reverses Cigarette Smoke-Induced Urocystic Epithelial-Mesenchymal Transition via Suppression of STAT3 Phosphorylation in SV-HUC-1-Immortalized Human Urothelial Cells.

PURPOSE: Bladder cancer is a malignant tumor of the urinary tract, and cigarette smoke (CS) is closely related to tumorigenesis. Resveratrol, a plant-derived bioactive nutrient, possesses multiple anticancer effects. However, the mechanism of CS-induced tumorigenesis is still not clear. The role of resveratrol in CS-meditated bladder cancer development has not been reported. METHODS: MTT assay showed the toxicity of cigarette smoke extract (CSE) on the cell viability of SV-HUC-1 cells. Western blotting detected the expression levels of related proteins. Transwell migration or invasion assay evaluated the capacity of cell migration or invasion after treatment. Wound-healing assay revealed the effect of cell migratory capacity. The cell cycle was detected by flow cytometry. RESULTS: Our study demonstrated that CSE-triggered epithelial-mesenchymal transition (EMT) in SV-HUC-1-immortalized human urothelial cells via the STAT3/TWIST1 pathway. Furthermore, the results showed resveratrol effectively inhibited STAT3 phosphorylation, thus reversed EMT triggered by CSE. Meanwhile, the cell proliferation was also suppressed. CONCLUSION: In conclusion, inhibition of the STAT3 in CSE-induced EMT on bladder cancer may be a promising cancer treatment target for suppression by resveratrol.

Histological subtype is a significant predictor for inguinal lymph node metastasis in patients with penile squamous cell carcinoma.

The present study aimed to investigate the relationship between histopathological subtype and the probability of inguinal lymph node metastasis (ILNM) in patients with penile squamous cell carcinoma (PSCC). The clinical records of 198 consecutive patients with PSCC were analyzed retrospectively. Primary lesions were reevaluated according to the 2016 World Health Organization (WHO) histopathological classification. We retrieved the clinicopathological factors from the medical records including age, clinical lymph node stage, pathological tumor stage, lymphatic invasion, and nerve invasion. Uni- and multivariate logistic regression analyses were used to explore the risk factors of ILNM. Multivariate analyses identified clinical lymph node stage (P = 0.000), pathological tumor stage (P = 0.016), histologic grade (P = 0.000), and risk group of histological subtypes (P = 0.029) as independent predictors for ILNM. Compared with the low-risk group of PSCC subtypes, the intermediate- (HR: 3.66, 95% CI: 1.30-10.37, P = 0.021) and high-risk groups (HR: 28.74, 95% CI: 2.37-348.54, P = 0.008) were significantly associated with ILNM. In conclusion, the histopathological subtype of the primary lesion is a significant predictor for ILNM in patients with PSCC.

Giant adrenal germ cell tumour in a 59-year-old woman.

Adrenal germ cell tumour is very rare. We report a case of a 59-year-old woman who presented with right flank discomfort. The laboratory examinations were normal and the chest computed tomography (CT) showed right pleural effusion. The abdominal CT scan revealed a large mass on the right adrenal gland. The patient underwent an adrenalectomy. Histopathologic examination and immunohistochemical findings were consistent with mixed germ cell tumour. Three months later following the operation, the patient was admitted to our hospital again with chest tightness and shortness of breath. The chest CT showed right pleural effusion recurrence and enlargement of mediastinal lymph nodes and right hilar lymph nodes. The patient had right supraclavicular lymphadenectasis on physical examination. Fine needle aspiration cytology from the supraclavicular lymph nodes showed groups of malignant tumour cells. The patient died within 6 months postoperatively. In this case, the lymph node pathway played an important role in the metastatic procedure.

Metastasis to the proximal ureter from prostatic adenocarcinoma: A rare metastatic pattern.

Prostate cancer is one of the most common male malignancies, but it rarely metastasizes to the proximal ureter. We report a case of a 76-year-old man who presented with flank pain and lower urinary tract symptoms. Abdominal computed tomography scan revealed multiple filling defects at the middle of the left ureter, enlarged retroperitoneal lymph nodes, and probable psoas invasion. The patient underwent nephroureterectomy with excision of a cuff of bladder, and was found to have an adhesion between the middle part of left ureter and psoas intraoperatively. The pathological examination displayed positive immunohistochemical staining with prostate-specific antigen and prostate acid phostate, supporting the diagnosis of metastatic ureteral tumour from prostate cancer. In this case, periureteral soft tissue and ureteral muscular layer were infiltrated by metastatic tumour, whereas the mucosa was spared. The periureteral lymphatic pathway played an important role in the metastatic procedure of prostate cancer to the proximal ureter.