Role of GM-CSF in a mouse model of experimental autoimmune prostatitis.

The etiology of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is still unknown. Granulocyte macrophage colony-stimulating factor (GM-CSF) has been shown to play an important role in the development of autoimmune and inflammatory diseases. Here, we investigated the expression and function of GM-CSF in patients with CP/CPPS and in a mouse model of experimental autoimmune prostatitis (EAP). GM-CSF mRNA levels were detected in expressed prostatic secretions samples from patients with CP/CPPS and in prostate tissue from a mouse model of EAP. The expression of GM-CSF receptor in mouse prostate and dorsal root ganglia were determined using PCR and immunohistochemistry. Behavioral testing and inflammation scoring were performed to evaluate the role of GM-CSF in disease development and symptom severity of EAP using GM-CSF knockout mice. mRNA levels of putative nociceptive and inflammatory markers were measured in the prostate after the induction of EAP. Elevated GM-CSF mRNA levels were observed in expressed prostatic secretions samples from patients with CP/CPPS compared with healthy volunteers. GM-CSF mRNA was also significantly increased in prostate tissue of the EAP mice model. The expression of GM-CSF receptors was confirmed in mouse prostate and dorsal root ganglia. GM-CSF knockout mice showed fewer Infiltrating leukocytes and pain symptoms after the induction of EAP. Deletion of GM-CSF significantly diminished EAP-induced increases of chemokine (C-C motif) ligand 2, chemokine (C-C motif) ligand 3, and nerve growth factor mRNA expression. The results indicated that GM-CSF plays a functional role in the pathogenesis of EAP. GM-CSF may function as a signaling mediator for both inflammation and pain transduction in CP/CPPS.

Amyloid precursor protein is overexpressed in bladder cancer and contributes to the malignant bladder cancer cell behaviors.

OBJECTIVES: To investigate the expression of amyloid precursor protein in bladder cancer, and to study its role in malignant bladder cancer cell behaviors. METHODS: Immunohistochemistry and western blotting analysis were used to detect the amyloid precursor protein level in bladder cancer tissues and cell lines. The effect of amyloid precursor protein on bladder cancer cell proliferation, migration, invasion and cell cycle was evaluated by using small interfering ribonucleic acid. The levels of RAS, RAF, MEK, phosphorylated MEK, extracellular regulated protein kinases, phosphorylated extracellular regulated protein kinases, protein kinase B and phosphorylated protein kinase B were determined by western blot after amyloid precursor protein knockdown. The effect of amyloid precursor protein on the extracellular regulated protein kinases pathway was further evaluated using extracellular regulated protein kinases pathway agonist, ceramide C6. RESULTS: The expression of amyloid precursor protein was significantly increased in the human bladder cancer tissues compared with matched normal bladder tissues. The overexpression of amyloid precursor protein was significantly associated with tumor stage, tumor size, histological grade and lymph node metastasis. The proliferation, migration and invasion of human bladder cancer cells were significantly inhibited by the silencing of amyloid precursor protein. In addition, knockdown of amyloid precursor protein arrested the cell cycle progression of bladder cancer cells in the G2/M phase. Mechanistic analysis showed that knockdown of amyloid precursor protein significantly decreased the phosphorylation of extracellular regulated protein kinases. Ceramide C6 could rescue the malignant bladder cancer cell behaviors inhibited by the silencing of amyloid precursor protein. CONCLUSIONS: Amyloid precursor protein is upregulated in bladder cancer, and might be closely associated with bladder cancer cell growth and survival. Amyloid precursor protein could potentially be used as a therapeutic target for bladder cancer treatment.

An uncommon manifestation of paraneoplastic cerebellar degeneration in a patient with high grade urothelial, carcinoma with squamous differentiation: A case report and literature review.

BACKGROUND: Paraneoplastic neurological syndromes (PNS) are rare disorders associated with malignant tumours, which are triggered by autoimmune reactions. Paraneoplastic cerebellar degeneration (PCD) is the PNS type most commonly associated with ovarian and breast cancer. Two bladder cancers manifesting in PCD were previously reported. However, the cancers in these cases had poor outcomes. CASE PRESENTATION: Here, we present a 68-year old man with history of high-grade papillary urothelial carcinoma of the bladder. The patient suffered from persistent cerebellar ataxia accompanied by bladder cancer recurrence five months after transurethral resection of the bladder tumour (TURBt). Laboratory screening for the specific antibodies of paraneoplastic neurological syndromes revealed no positive results. Symptoms were not remitted after a 7-day-course of high-dose glucocorticoid therapy. To our surprise, the patient recovered fully after laparoscopic radical cystectomy. Postoperative pathology revealed that surgical specimens were urothelial carcinoma in situ (CIS) and squamous cell carcinoma of the bladder. The patient remained asymptomatic and there was no evidence of recurrence after the followup period of 11 months. CONCLUSION: To our knowledge, this is the third report of PCD in a patient with bladder cancer. This case showed that tumour resection cured the PCD. To assist clinical evaluation and management, literature regarding basic PNS characteristics and bladder cancers was reviewed.

Increased expression of miR-222 is associated with poor prognosis in bladder cancer.

BACKGROUND: MicroRNA-222 (miR-222) has been shown to play a potential oncogenic role in bladder cancer. The aim of this study was to evaluate the expression of miR-222 in bladder cancer and its potential relevance to clinicopathological characteristics and patient survival. METHODS: Surgical specimens of cancer tissue and adjacent normal tissue were obtained from 97 patients with bladder cancer. The relative expression levels of miR-222 in the cancer and the normal adjacent tissue were measured by quantitative reverse-transcriptase PCR. We analyzed their correlation with clinicopathological parameters and prognostic value. RESULTS: The expression level of miR-222 was significantly higher in tumor tissues than in corresponding non-cancerous tissues (5.46 ± 1.45 versus 1.92 ± 0.65, P < 0.0001), and a high expression of miR-222 was found to be significantly associated with tumor grade (P = 0.003) and tumor stage (P = 0.005). The miR-222 expression level was classified as high or low in relation to the median value (cutoff value = 5.15). Kaplan-Meier analysis showed that patients with higher levels of miR-222 had significantly poorer survival than those with lower expression of this miRNA in patients, with a 5-year overall survival of 29.53% and 52.75%, respectively (P = 0.0034). In the multivariate Cox proportional hazards analysis, which included miR-222 level, tumor grade, tumor stage, and tumor number, high miR-222 expression was independently associated with poor survival (P < 0.001; hazard ratio 6.17; 95% CI 2.33 to 10.39). CONCLUSION: miR-222 overexpression is involved in the poor prognosis of bladder cancer and can be used as a biomarker for selection of cases requiring special attention.

Comparison of flexible ureteroscopy and mini-percutaneous nephrolithotomy in the treatment for renal calculi larger than 2 cm: a matched-pair analysis.

To compare the effectiveness and safety of flexible ureteroscopy and mini-percutaneous nephrolithotomy for renal calculi > 2 cm and perform subgroup analysis of stone length and age. Patients received mini-percutaneous nephrolithotomy or flexible ureteroscopy in Qilu Hospital of Shandong University from 2016.01 to 2021.03 with renal calculi > 2 cm were retrospectively analyzed. Propensity score matching was performed to get comparable patients. The postoperative hospital days, operation time, complication rate, and stone free rate were compared. The age and stone length were analyzed by subgroup. 162 in 313 patients were finally included. Each group had 81 cases. Outcomes such as intraoperative transfusion, stone free rate show no difference either. Flexible ureteroscopy had shorter postoperative hospital days (3.2 days vs 7.2 days, P < 0.001) and fewer complications (9, 11.1% vs 25, 30.9%, P = 0.002) compared to mini-percutaneous nephrolithotomy. The postoperative hospital days, and complication of the flexible ureteroscopy were significantly lower than those in the mini-percutaneous nephrolithotomy for renal stones ≤ 2.5 cm; when the stone length > 2.5 cm, the stone free rate of flexible ureteroscopy was lower than that of the mini-percutaneous nephrolithotomy group, but not statistically significant. The complications of flexible ureteroscopy in the young group (18-39 years old) were significantly lower than those in the mini-percutaneous nephrolithotomy group. For 2-2.5 cm renal stones, flexible ureteroscopy can achieve a similar stone free rate with shorter hospital stay, and lower complications. For larger stones, flexible ureteroscopy performed poorly. Flexible ureteroscopy may be a better option for younger patients with fewer complications.

Comparison of flexible ureteroscopy and mini-percutaneous nephrolithotomy in the treatment for multiple nephrolithiasis.

Objective: To compare the outcomes of flexible ureteroscopy and mini-percutaneous nephrolithotomy in the treatment for multiple nephrolithiasis in 1-2 cm size. Methods: The clinical data of patients with multiple renal calculi in the range of 1-2 CM who underwent flexible ureteroscopy lithotripsy and percutaneous nephrolithotomy in Qilu Hospital of Shandong University from January 2016 to March 2021 were retrospectively collected and matched using propensity score matching. Then a subgrouping of the number of stones was performed. Patients were divided into Group A and Group B according to their stone numbers. Patients with no statistically significant differences in baseline data were matched to compare the safety and efficacy of the two procedures. Results: A total of 210 patients with clinical data were collected, and the patients' baseline data were not comparable, and 142 patients were finally included in the study after propensity score matching. There was no statistical difference in baseline data between the two groups of patients. The postoperative hospital days (3.00, 2.00 vs. 7.00, 3.00, P < 0.001), operation time (90.00, 50.00 vs. 110.00, 53.00, P = 0.018), complications (6, 6.8% vs. 14, 25.9%, P = 0.001) of patients in flexible ureteroscopy group %, P = 0.001) was significantly lower than that in the percutaneous nephrolithotomy group. There was no significant difference in stone clearance rate between the two groups (76, 86.4% vs. 42, 77.8%, P = 0.185). When the number of stones was no more than 3, the operation time (85.00, 49.00 vs. 110.00, 53.00, P = 0.005) and complications (2, 4.2% vs. 11, 29.7%, P = 0.001) of f-URS were significantly less than those of mPCNL, but when the number of stones was more than 3, there was no significant difference between the two operations. Conclusion: For multiple nephrolithiasis within 1-2 CM, when the number of stones does not exceed 3, flexible ureteroscopy can achieve the same stone clearance rate as percutaneous nephrolithotomy, while having shorter post-operation days, operative time and fewer complications. When the number of stones is more than 3, there are no significant difference between two operations.

Safety and efficacy of extracorporeal shock wave lithotripsy vs. flexible ureteroscopy in the treatment of urinary calculi: A systematic review and meta-analysis.

Objective: This study aims to compare the safety and efficacy of extracorporeal shock wave lithotripsy (SWL) and flexible ureteroscopy lithotripsy (f-URS) in treating urinary tract stones. Methods: We systematically searched PubMed, Embase, and Cochrane for literature comparing SWL with f-URS. The primary outcomes we focused on were stone-free rate (SFR) and complications; the secondary outcomes were operation time, hospital stay, retreatment rate, number of sessions, and auxiliary procedures rate. We used ReviewManager version 5.4.1 and STATA version 14.2 for meta-analysis. Results: Seventeen studies with a total of 2,265 patients were included in the meta-analysis, including 1,038 patients in the SWL group and 1,227 patients in the f-URS group. The meta-analysis indicated that patients in the f-URS group had higher SFR than those in the SWL group [odds ratio (OR): 2.00, 95% confidence interval (CI): 1.29-3.12, p = 0.002]. In addition, we found no significant difference in complications (OR: 1.08, 95% CI: 0.85-1.37) between the two treatments. Also, we found that the retreatment rate and the auxiliary procedure rate in the f-URS group were significantly lower than those in the SWL group (OR: 0.08, 95% CI: 0.02-0.24, p < 0.00001; OR: 0.30, 95% CI: 0.11-0.83, p = 0.02). Moreover, the number of sessions in the f-URS group was significantly lower than that in the SWL group [mean difference (MD): -1.96, 95% CI: -1.55 to -0.33, p = 0.003]. However, the operation time and hospital stay in the f-URS group were significantly longer than those in the SWL group (MD: 11.24, 95% CI: 3.51-18.56, p = 0.004; MD: 1.14, 95% CI: 0.85-1.42, p < 0.00001). Conclusion: For 1-2-cm urinary stones, f-URS can achieve a higher SFR than SWL while having a lower retreatment rate, number of sessions, and auxiliary procedure rate. For urinary stones <1 cm, there was no significant difference in SFR between SWL and f-URS groups. The SWL group has a shorter operative time and hospital stay than the f-URS group.

Anticancer Properties and Mechanisms of Singly-Protonated Dehydronorcantharidin Silver Coordination Polymer in a Bladder Cancer Model.

Bladder cancer is the most common malignant urinary system tumor. Chemotherapy is frequently used as a treatment regimen for patients with bladder cancer, however, new and effective drugs for bladder cancer need to be developed. The present study examined the effects and mechanisms of Ag-SP-DNC, a silver and singly-protonated dehydronorcantharidin complex, on bladder cancer in vitro and in vivo. It was identified that Ag-SP-DNC suppressed cell proliferation and induced apoptosis in bladder cancer cells in vitro, a suppression associated with G0/G1 phase arrest and elevated intracellular reactive oxygen species (ROS) levels. Furthermore, Ag-SP-DNC enhanced the cleaved caspase-3 levels, disrupted the mitochondrial transmembrane potential balance, and induced intracellular calcium overload. The Ag-SP-DNC-induced bladder cancer cell apoptosis was significantly decreased following treatment with a broad caspase inhibitor, zVAD-fmk. In addition, treatment of MB49 tumor-bearing mice with Ag-SP-DNC significantly inhibited tumor growth and decreased the anti-apoptosis and cell cycle promotion protein levels in the tumor. The results of the present study suggested that Ag-SP-DNC elicits a strong anticancer effect against bladder cancer, and can therefore be used as a promising treatment for bladder cancer.

Bulbocavernosus reflex to stimulation of prostatic urethra in patients with lifelong premature ejaculation.

INTRODUCTION: Lifelong premature ejaculation (LPE) is a prevalent sexual dysfunction among men, while its precise pathologic mechanisms have remained poorly understood. AIM: In our study, the correlation between excitability of bulbocavernosus reflex (BCR) to stimulation of the prostatic urethra and LPE was studied. METHODS: Twenty normal potent male volunteers and 42 patients with LPE were studied by inserting a specially designed Foley catheter with two electrodes mounted on its distal surface (intraurethral catheter electrode) into bladder to evoke the BCR to stimulation of prostatic urethra. Also, sensitivity of glans penis to electrical stimulation was detected by two surface electrodes. MAIN OUTCOME MEASURES: Sensory thresholds of BCR to stimulation of prostatic urethra, thresholds to evoke stable BCR, latencies of BCR, and sensory thresholds of glans penis to electrical stimulation. RESULTS: The mean sensory thresholds of BCR to stimulation of prostatic urethra, thresholds to evoke stable BCR, latencies of BCR, and sensory thresholds of glans penis were 12.38±3.71 mA (0.2 ms in duration, 1 Hz), 23.81±5.55 mA (0.2 ms, 1 Hz), 70.48±6.33 ms, and 11.89±2.26 mA (0.04 ms in duration,3 Hz) in the patients with LPE, respectively, and were 18.20±2.68 mA (0.2 ms, 1 Hz), 34.76 ± 4.15 mA (0.2 ms, 1 Hz), 71.20±5.77 ms, and 14.16±1.94 mA (0.04 ms, 3 Hz) in the normal potent men, respectively (mean±SD). Statistically significant differences were seen regarding the sensory thresholds of BCR to stimulation of prostatic urethra, the thresholds to evoke stable BCR and the sensory thresholds of glans penis between the two groups (P<0.001). No statistically significant differences were seen regarding the latencies of BCR between the two groups (P>0.05). CONCLUSIONS: Patients with LPE might have hyperexcitable BCR to stimulation of prostatic urethra, which is probably one of the important factors for its etiology.

Bone marrow mesenchymal stem cells induce angiogenesis and promote bladder cancer growth in a rabbit model.

OBJECTIVES: To investigate the effect of mesenchymal stem cells (MSCs) in the process of tumor development and the possibility of MSCs differentiating into vascular endothelial cells in the tumor microenvironment. MATERIAL AND METHODS: Twenty male New Zealand rabbits were randomly divided into 2 groups: a test group and a control group. MSCs were isolated and cultured by bone marrow cell adherence. The bladder tumor models were built by embedding a VX2 mass in swelled bladder mucosa in all of the rabbits (n = 20). One week later, 4',6-diamidino-2-phenylindole-labeling MSCs were transplanted into tumor tissue in the test group (n = 10). Culture medium was injected into the tumor tissue of the control group (n = 10). The maximum diameter of the tumor mass was measured by ultrasound at 2 and 4 weeks after the VX2 tumor mass was embedded. All animals were sacrificed at 4 weeks. The double labeling immunofluorescence for CD146 was performed to reveal whether engrafted cells can differentiate into vascular endothelial cells. Vascular density was compared between the 2 groups. RESULTS: There was no significant difference in the maximum diameters of the tumor masses between the 2 groups at 2 weeks (test group 0.77 +/- 0.15 cm vs. control group 0.71 +/- 0.15 cm, p > 0.05). The maximum diameters appeared larger in the test group at 4 weeks (test group 3.82 +/- 0.94 cm vs. control group 2.28 +/- 0.54 cm, p < 0.05). Immunofluorescence studies revealed some engrafted MSCs expressing a vascular endothelial cell phenotype (CD146). Furthermore, vascular density was augmented in the test group in comparison to the control group (10.1 +/- 0.70/0.2 mm(2) vs. 8.24 +/- 0.81/0.2 mm(2), p < 0.05). CONCLUSIONS: Engrafted MSCs can differentiate into vascular endothelial cells and contribute to angiogenesis in the tumor microenvironment, which may be the major pathway of promoting tumor growth.

Vitamin C Inhibits Metastasis of Peritoneal Tumors By Preventing Spheroid Formation in ID8 Murine Epithelial Peritoneal Cancer Model.

High mortality is associated with exclusively metastasis within the peritoneal cavity among patients with epithelial ovarian cancer that is the most lethal gynecologic cancer. There is an unmet need to develop more effective therapies to prevent metastasis of peritoneal cancer. Multicellular spheroid formation, during which cancer cells migrate and adhere to tumor-associated macrophages, is a critical step of ovarian cancer metastasis. Here, we showed that vitamin C inhibited spheroid formation and metastasis in ID8 ovarian cancer-bearing mice. We further found that vitamin C treatment decreased the levels of M2 macrophages in tumor nodules and suppressed the epithelial-mesenchymal transition (EMT). In vitro studies revealed that vitamin C inhibited proliferation, arrested cell cycle, attenuated migration, and prevented the spheroid formation of ID8 ovarian cancer cells. Vitamin C induced apoptosis of ID8 cells, which was confirmed by membrane potential collapse, cytosolic calcium overload, ATP depletion, and caspase-3 activation in vitamin C-treated cells. Intriguingly, vitamin C treatment caused striking morphological change and apoptosis of macrophages. The presented proof of concept study strategically identifies new anticancer mechanisms of vitamin C.

Does preoperative urine culture still play a role in predicting post-PCNL SIRS? A retrospective cohort study.

There is controversy regarding the predicting value of preoperative urine culture for post-percutaneous nephrolithotripsy (PCNL) infection. The purpose of our study was to re-evaluate the importance of preoperative urine culture for developing post-PCNL systemic inflammatory response syndrome (SIRS) in China. A total of 303 patients undergone PCNL from March 2012 to January 2018 were recruited. Urine tests, urine cultures and the perioperative data were prospectively recorded and analyzed. 95 patients (31.4%) were identified with positive preoperative urine cultures. Female patients had significantly higher rate of urine cultures positivity than that in male patients (42.9% vs. 21.5%, p < 0.01). Escherichia coli was the most common organism (56 cases, 58.9%) in patients with positive urine cultures and 35.7% of E. coli-positive patients developed SIRS after PCNL. Even with intensive perioperative prophylaxis, patients with positive urine cultures had a higher rate of post-PCNL SIRS than those patients with negative urine cultures (p = 0.043). On multivariable analysis, preoperative positive urine cultures (OR 1.943, 95% CI 1.11-3.39, p = 0.019), preoperative neutrophils (OR 1.228, 95% CI 1.07-1.41, p = 0.003), intraoperative pyonephrosis (OR 3.37, 95% CI 1.44-9.71, p = 0.01) and postoperative hospitalization (OR 1.154, 95% CI 1.05-1.28, p = 0.005) were independent risk factors for postoperative SIRS. These results demonstrated that preoperative urine culture still played a role in predicting post-PCNL SIRS, but it was unable to prevent the occurrence of SIRS even with intensive perioperative prophylaxis based on urine culture results. Further studies are required to explore the predicting role of advanced preoperative bacterial detecting techniques in post-PCNL infectious complications.

[Role of bulbocavernosus reflex to stimulation of prostatic urethra in pathologic mechanism of primary premature ejaculation].

OBJECTIVE: Primary premature ejaculation (PPE) is a prevalent sexual dysfunction among men while its precise pathologic mechanism has remained poorly understood. In current study the correlation between excitability of bulbocavernosus reflex (BCR) to stimulation of prostatic urethra and primary premature ejaculation was studied. METHODS: Forty-two patients with PPE and 20 normal potent male volunteers were studied by inserting a specially designed Foley catheter with two electrodes mounted on its distal surface (intraurethral catheter electrode) into bladder to evoke the BCR to stimulation of prostatic urethra to record the sensory thresholds of BCR to stimulation of prostatic urethra, thresholds to evoke stable BCR and latencies of BCR. Also the sensitivity of glans penis to electrical stimulation was detected by two surface electrodes. RESULTS: The mean sensory thresholds of BCR to stimulation of prostatic urethra, thresholds to evoke stable BCR, latencies of BCR and sensory thresholds of glans penis were (18.2 +/- 2.7) mA (0.2 ms in duration, 1 Hz), (34.8 +/- 4.2) mA (0.2 ms, 1 Hz), (71.2 +/- 5.8) ms and (14.2 +/- 1.9) mA (0.04 ms in duration, 3 Hz) in normal potent men respectively and were (12.4 +/- 3.7) mA (0.2 ms, 1 Hz), (23.8 +/- 5.6) mA (0.2 ms, 1 Hz), (70.5 +/- 6.3) ms and (11.9 +/- 2.3) mA (0.04 ms, 3 Hz) in patients with PPE respectively. Statistically significant differences were seen regarding the sensory thresholds of BCR to stimulation of prostatic urethra, the thresholds to evoke stable BCR and the sensory thresholds of glans penis between two groups (all P < 0.01). No statistically differences were seen regarding the latencies of BCR between two groups (P > 0.05). CONCLUSION: Patients with PPE have hyperexcitable BCR to stimulation of prostatic urethra. It is probably one of the important etiological factors. Moreover the findings may provide new therapeutic modalities of PPE.

Synchronous primary sigmoid colon cancer and primary thyroid cancer followed by a malignant tumor of the kidney: Case report of multiple primary cancer and review of the literature.

Multiple primary cancer (MPC) is relatively rare. With the development of diagnostic and anti-cancer therapeutic techniques, the incidence of MPC is rising annually. However, the incidence of triple or quadruple cancers in a single patient remains low. In this report, the case of a 58-year-old male with triple malignant cancer is outlined. Synchronous sigmoid colon cancer and thyroid cancer were diagnosed in May 2015; on subsequent re-examination, metastasis to the liver and a malignant kidney tumor were also identified. The diagnosis was established via computed tomography (CT), Positron emission tomography-CT (PET-CT) and other laboratory examination results, including analysis of tumor markers and liver function, and was confirmed by pathological diagnosis. The patient underwent radical surgery and standardized chemotherapy. Through literature review, the definition, characteristics, classification, incidence, possible causes of and treatment strategies for MPC were more clearly understood. In addition, immunohistochemical staining of integrin αvß6 was performed on patient tissue specimens, where integrin αvß6 expression was confirmed in cancer of the colon, thyroid and liver, as a result of colonic metastasis. Therefore, the involvement of integrin αvß6 in the malignant progression of MPC was hypothesized, which may aid the investigation of MPC etiology in the future.

Role of APLP2 in the prognosis and clinicopathology of renal cell carcinoma.

Identifying diagnostic and prognostic biomarkers is crucial for improved guidance of the treatment of renal cell carcinoma (RCC). Amyloid ß precursor-like protein 2 (APLP2) has been determined to serve an important role in the progression of a number of cancer types. However, the expression and significance of APLP2 in RCC remains unknown. In the present study, it was determined that the expression of APLP2 protein (n=10) and mRNA (n=8) expression was significantly decreased in clear cell RCC (CCRCC) tissues compared with that in matched normal renal tissues. The expression level of APLP2 was significantly associated with high Fuhrman grade, high pT stage, and presence of distant metastasis and lymph node metastasis (P<0.05). Multivariate analysis demonstrated that the expression of APLP2 was a significant independent predictor of disease-specific survival in renal cell carcinoma (P=0.026). Notably, APLP2 expression was significantly associated with disease-specific survival (P<0.001). APLP2 may be used to potentially predict patient prognosis, and to guide clinical diagnosis and treatment in CCRCC.

A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue.

Primary human keratinocytes isolated from fresh skin tissues and their expansion in vitro have been widely used for laboratory research and for clinical applications. The conventional isolation method of human keratinocytes involves a two-step sequential enzymatic digestion procedure, which has been proven to be inefficient in generating primary cells from adult tissues due to the low cell recovery rate and reduced cell viability. We recently reported an advanced method to isolate human primary epidermal progenitor cells from skin tissues that utilizes the Rho kinase inhibitor Y-27632 in the medium. Compared with the traditional protocol, this new method is simpler, easier, and less time-consuming, and increases epithelial stem cell yield and enhances their stem cell characteristics. Moreover, the new methodology does not require the separation of the epidermis from the dermis, and, therefore, is suitable for isolating cells from different types of adult tissues. This new isolation method overcomes the major shortcomings of conventional methods and is more suitable for producing large numbers of epidermal cells with high potency both for laboratory and for clinical applications. Here, we describe the new method in detail.

Identification of hub genes and pathways associated with bladder cancer based on co-expression network analysis.

The aim of the present study was to identify hub genes and signaling pathways associated with bladder cancer (BC) utilizing centrality analysis and pathway enrichment analysis. The differentially expressed genes (DEGs) were screened from the ArrayExpress database between normal subjects and BC patients. Co-expression networks of BC were constructed using differentially co-expressed genes and links, and hub genes were investigated by degree centrality analysis of co-expression networks in BC. The enriched signaling pathways were investigated by Kyoto Encyclopedia of Genes and Genomes database analysis based on the DEGs. The hub gene expression in BC tissues was validated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. A total of 329 DEGs were screened, including 147 upregulated and 182 downregulated genes. The co-expression network constructed between BC and normal controls consisted of 182 nodes and 434 edges, and the two genes in each gene pair were differentially co-expressed genes. Centrality analysis of co-expression networks suggested that the top 5 hub genes with high degree included lectin, galactoside-binding, soluble, 4 (LGALS4), protein tyrosine phosphatase, receptor type N2 (PTPRN2), transmembrane protease, serine 11E (TMPRSS11E), tripartite motif containing 31 (TRIM31) and potassium voltage-gated channel subfamily D member 3 (KCND3). Pathway analysis revealed that the 329 DEGs were significantly enriched in 5 terms (cell cycle, DNA replication, oocyte meiosis, p53 signaling pathway and peroxisome proliferator-activated receptor signaling pathway). According to RT-qPCR and western blot analysis, 4/5 hub genes were significantly expressed, including LGALS4, PTPRN2, TMPRSS11E, TRIM31; however, KCND3 was not significantly expressed. In the present study, 5 hub genes were successfully identified (LGALS4, PTPRN2, TMPRSS11E, TRIM31 and KCND3) and 5 biological pathways that may be underlying biomarkers for early diagnosis and treatment associated with bladder cancer were revealed.

The role of c-Met in prognosis and clinicopathology of renal cell carcinoma: Results from a single-centre study and systematic review.

BACKGROUND AND OBJECTIVES: The c-Met proto-oncogene pathway plays an important role in the progression of various cancers. However, the effect of the c-Met pathway on renal cell carcinoma (RCC) remains controversial. We decided to clarify the role of c-Met in prognosis and clinicopathology of RCC. METHODS: A total of 10 pairs of tumour and adjacent tissues were obtained from patients with primary RCC between 2013 and 2014 and tissue microarrays to assess c-Met expression in tumour tissues from 90 patients with RCC by Western blot and immunohistochemical staining. We also presented a meta-analysis to explore the correlation between c-Met and pathological grade and stage of RCC. The two-tailed Pearson's χ2 and Fischer exact tests were used to compare categorical variables. Multivariate analysis was performed using the multivariate Cox proportional hazards model. RESULTS: C-Met protein levels were increased in 8 of 10 RCC tissue samples compared with their adjacent normal tissue and c-Met expression levels were positively associated with a high nuclear grade (P = 0.008) and pT stage (P = 0.002). Multivariate analysis showed that a high expression of c-Met was an independent predictor of disease-specific survival (P = 0.017). A meta-analysis found that increased c-Met expression in RCC tissues was closely correlated with high tumour grade (P<0.001) and high pT stage (P = 0.001). Most importantly, c-Met expression was significantly correlated with disease-specific survival (P<0.001). CONCLUSIONS: Because c-Met is strongly associated with pathological grade, stage and disease-specific survival, c-Met levels may have potential to predict patient prognosis and to guide clinical diagnosis and treatment.