Bladder cancer tissue-derived exosomes suppress ferroptosis of T24 bladder cancer cells by transporting miR-217.

It has been reported that miR-217 can inhibit the oncogenic activity and progression of bladder cancer (BCa) cells, but it has not been explored whether miR-217 is involved in the regulation of ferroptosis. In the present study, RNA transfection, real-time PCR, flow cytometry, Western blotting assays, immunofluorescence and ELISA were performed to explore the effects and mechanisms of miR-217 in BCa tissue-derived exosomes. We found that extracellular fluid from bladder cancer tissue promoted the growth and miR-217 expression of T24 cells and inhibited ferroptosis. MiR-217 was confirmed to inhibit ferroptosis in bladder cancer cells by RNA interference and functional assays. By cell membrane fluorescence probe (CM-Dil) labeling, inhibiting exosome secretion by GW4689 and exosome extraction, we determined that BCa tissue-derived exosomes transport miR-217 into T24 cells. Culture of T24 cells with extracellular fluid after RNA interference showed that exosomes carrying miR-217 derived from BCa tissues inhibited ferroptosis of T24 cells. We conclude that bladder cancer tissue-derived exosomes inhibit ferroptosis of T24 bladder cancer cells by transporting miR-217. The results of our study provide a new insight into the progression of bladder cancer.

Bladder mesenchymal stromal cell-derived exosomal miRNA-217 modulates bladder cancer cell survival through Hippo-YAP pathway.

BACKGROUND: Donor cell-derived exosomes regulate recipient cell functions. The aim of this study was to investigate the effect of human normal bladder stromal cell (hBSC) derived exosomal miR-217 on bladder cell cancer proliferation and migration. METHODS: Human BSCs were transfected with miR-217 mimic or inhibitor and hBSC-derived exosomes were isolated. Human bladder cancer cell lines (T24 and 5367) were co-cultured with hBSC-derived exosomal miR-217 mimic or inhibitor. Proliferation, migration, and apoptosis of the bladder cancer cells were assessed by Edu assay, Transwell migration assay, and Annexin V assay. RESULTS: Expression of miR-217 was significantly higher in the T24 and 5367 cell lines (P < 0.01). Exosomal miR-217 mimic enhanced proliferation and migration of T24 and 5367 cells, but inhibited apoptosis of the cells (P < 0.01); in contrast, exosomal miR-217 inhibitor suppressed proliferation and migration but stimulated apoptosis of the two cancer cell lines (P < 0.01). Moreover, exosomal miR-217 mimic stimulated YAP and its target proteins including Cyr61, CTGF, and ANKRD1 (P < 0.01), and in contrast, exosomal miR-217 inhibitor suppressed YAP and its target proteins (P < 0.01). CONCLUSION: These findings suggested that hBSC-derived exosomal miR-217 may act as oncogene in bladder cancer cells, and that Hippo-YAP signaling pathway maybe the target for miR-217 in the bladder cancer cell lines.

CircRNA-100284 activates aurora kinase B by inducing methylation of HSP70 via microRNA-217 to promote proliferation of bladder cancer cells.

OBJECTIVE: The malignant transformation of normal bladder cells (SV-HUC-1) was induced by arsenite to explore the possible mechanism of circRNA-100284 influencing bladder cancer cell proliferation. METHODS: Normal bladder SV-HUC-1 cells were cultured with 2 µM arsenite to induce malignant transformation. After 0, 3, 6, 12, and 24 h of culture, the expression level of circRNA-100284 in cells was detected by quantitative real-time PCR. Western blotting assays were used to detect the expression levels of EZH2 and cyclin-D1 proteins in cells treated with different media. Cell cycle was analyzed by flow cytometry. In addition, through cell transfection and CCK-8 experiments, the effect and mechanism of circRNA-100284 targeting microRNA-217 on proliferation was determined. The interaction between HSP70 methylation and Aurora-B was determined by Western blotting and immunoprecipitation experiments. RESULTS: With prolonged contact time with arsenite, the expression level of circRNA-100284 in cells increased continuously (P < 0.05). Western blotting assays showed that the expression levels of EZH2 and cyclin-D1 proteins in arsenite-transformed cells increased. Flow cytometry and CCK-8 showed that circRNA-100284 accelerated cell cycle transition and cell proliferation through miR-217. Finally, after culturing human bladder cancer T24 cells, combined with immunoprecipitation and in vitro kinase experiments, it was found that K561- dimethyl HSP70 activated Aurora-B, thus promoting the proliferation of bladder cancer cells. CONCLUSION: CircRNA-100284 activates aurora kinase B by inducing methylation of HSP70 via microRNA-217 to promote the proliferation of bladder cancer cells.

Renal medullary carcinoma masquerading as renal infection: a case report.

BACKGROUND: Renal medullary carcinoma is a rare and aggressive tumor and often seen in young adults with sickle cell hemoglobinopathies. CASE PRESENTATION: We report a case of renal medullary carcinoma in a 29-year old male patient with an occupying renal lesion who presented with fever, flank pain and hematuria. The patient received intensive antibiotics treatment, but no improvement was seen. The symptoms disappeared after laparoscopic radical left nephrectomy. Postoperative pathological study showed that the mass was renal medullary carcinoma. CONCLUSIONS: Our case suggests that renal medullary carcinoma should be considered in differential diagnoses of patients with occupying renal lesions who have fever of unknown origin.

Intraoperative Vaginal Perforation During Various Mid-Urethral Sling Procedures Treating Female Stress Urinary Incontinence.

BACKGROUND: Incidence of intraoperative vaginal perforation is generally considered to be low but varies among different procedures. Vaginal perforation could not only prolong the surgeries and aggravate surgical trauma but also result in postoperative discomfort or even a second surgery. METHOD: Vaginal perforation, vaginal epithelial perforation, vaginal wall perforation, vaginal penetration, urinary incontinence were searched in PubMed, Cochrane, Embase database to identify the qualified clinical trial and relevant literature sources were also searched. RESULTS: A total of 9223 cases of from 33 trials from literatures and 387 cases from our own trail were analyzed, which provided detailed data on intraoperative vaginal perforation. Incidence of intraoperative vaginal perforation during mid-urethral sling surgery treating stress urinary incontinence was generally low, which was 1.56%. Incidence of intraoperative vaginal perforation during transobturator (TOR) procedure was higher than that during retropubic (RPR) procedure, which were 2.11% and 0.89% respectively. Incidence of intraoperative vaginal perforation during outside-to-inside TOR procedure like TOT and MONARC was higher than that during inside-to-outside TOR procedure like TVT-O, which were 2.74% and 1.52%, respectively. Incidence of intraoperative vaginal perforation during single-incision surgery like H-type TVT-SECUR reached 1.97%, while no report on U-type TVT-SECUR surgeries. CONCLUSION: The incidence of intraoperative vaginal during mid-urethral sling procedures for female stress urinary incontinence is fairly high. Vaginal perforation was more common in trans-obturator route (TOR) than retropubic route (RPR). In TOR route, it was less frequent in inside-to-outside procedure than outside-to-inside procedure. Surgery proficiency could also have an impact on this complication.

[Double Endobutto reconstituting coracoclavicular ligament combined with repairing acromioclavicular ligament at stage I for the treatment of acromioclavicular dislocation with Rockwood type III - V].

OBJECTIVE: To explore clinical efficacy of double Endobutto reconstituting coracoclavicular ligament combined with repairing acromioclavicular ligament in stage I in treating acromioclavicular dislocation with Rockwood type III - V . METHODS: From January 2010 to September 2013, 56 patients with Rockwood type III - V acromioclavicular dislocation were treated by operation, including 20 males and 36 femlaes, aged from 32 to 52 years old with an average of 38.5 years old. Twenty-five patients were on the left side and 31 cases on the right side. The time from injury to operation was from 3 to 14 days, averaged 7 days. All patients were diagnosed as acromioclavicular dislocation with Rockwood type III - V, and double Endobutto were used to reconstituting coracoclavicular ligament, line metal anchors were applied for repairing acromioclavicular ligament. Postoperative complications were observed, Karlsson and Constant-Murley evaluation standard were used to evaluate clinical effects. RESULTS: All patients were followed up from 8 to 24 months with average of 11 months. According to Karlsson evaluation standard at 6 months after operation,42 cases were grade A, 13 were grade B and 1 was grade C. Constant-Murley score were improved from (42.80±5.43) before operation to (91.75±4.27) at 6 months after operation. All items at 6 months after operation were better than that of preoperative items. Forty-eight patients got excellent results, 7 were moderate and only 1 with bad result. No shoulder joint adhesion, screw loosening or breakage were occurred during following up. CONCLUSION: Double Endobutto reconstituting coracoclavicular ligament combined with repairing acromioclavicular ligament in stage I for the treatment of acromioclavicular dislocation with Rockwood type III - V could obtain early staisfied clinical effects, and benefit for early recovery of shoulder joint function.

[Effect of tensile stress on human heel skin fibroblast proliferation in vitro].

OBJECTIVE: To observe the effect of tensile stress on human heel skin fibroblast proliferation in vitro, providing a theoretical basis for preventing the wound edge skin necrosis and nonunion after calcaneal fracture surgery. METHODS: Fibroblast cells were taken from lateral heel skin of a 40 year-old-man, then cultured and subcultured in vitro. After that, they were divided into three groups: 0 hours group, 6 hours group and 24 hours group and were tested by tensile stress testing. The levels of TGF-ß1 and IL-6 in nutrient fluid were measured. Transmission electron microscope and light microscope was applied for observe mitochondria and nucleus. RESULTS: Under 10% of the tensile stress, mitochondria decreased, the levels of TGF-ß1 and IL-6 in nutrient fluid were decreased and cell proliferation was inhibited gradually with time increasing. CONCLUSION: The human lateral heel skin in a long-time tensile stress state is an important cause of wound edge skin necrosis and nonunion after calcaneus fracture surgery.

Difference of adherence, proliferation and osteogenesis of mesenchymal stem cells cultured on different HA/ZrO2 composites.

OBJECTIVE: To study the adherence, proliferation and osteogenesis of mesenchymal stem cells (MSCs) cultured on different HA/ZrO2 composites. METHODS: The simplex and graded HA/ZrO2 compo-sites were prepared using dry-laid method. The surface topography of the composites was observed by scanning electron microscope (SEM). The MSCs were isolated from rabbits and cultured on experimental groups (simplex HA/ZrO2 composite, graded HA/ZrO2 composite, pure HA or pure ZrO2 coatings respectively) and control group (ordinary culture plate). Then, we observed the adherence, proliferation and osteogenesis of the MSCs, detected the cellular alkaline phosphatase (ALP) activities, extracted total RNA and detected the mRNA expression of collagen I, osteocalcin and osteopontin using the reverse transcription and polymerase chain reaction (RT-PCR) method. RESULTS: The SEM images confirmed that the surface of the simplex HA/ZrO2 composite was coverd by discon-tiguous HA layer with clear visualization of the partial ZrO2 matrix, while the surface of the graded HA/ZrO2 composite was fairly rough with porosity. X-ray diffraction showed that after high temperature sintering, the ZrO2 phase still remained, while the HA phase was transformed to beta-Ca3(PO4)2 , alpha-Ca3(PO4)2 and CaZrO3 phases on the surface of both composites. Cell culture indicated that the HA/ZrO2 composites supported cell attachment. Neither ALP expression nor mRNA expression of collagen I, osteocalcin or osteopontin from RT-PCR results showed significant deviation among four groups. CONCLUSION: Among these four composites, the graded HA/ZrO2 composite promotes the MSCs proliferation and the osteogenic differentiation to a certain extent.

[Clinical diagnosis and treatment of spontaneous urinary extravasation caused by primary lower ureteral tumor].

OBJECTIVE: To explore the clinical characteristics of spontaneous urinary extravasation caused by primary lower ureteral cancer. METHODS: Eight cases of spontaneous urinary extravasation caused by the obstruction of primary ureteral cancer from January 2005 to June 2010 from Department of Urology, Peking Union Medical College Hospital, were reported. There were 5 males and 3 females with an age range of 56-81 years old. Six cases presented with an onset of acute flank pains and 2 cases complained of flank discomforts with a lower fever. All cases had peripelvic fluid collection of varying levels on computed tomography (CT) scan and extravasation of contrast medium was found in delayed enhanced CT. The tumors of ureter were detected in 6 cases through preoperative imaging examinations while another 2 cases had no evidence of tumor. RESULTS: Unilateral nephroureterectomy was performed in 6 cases, including 5 cases undergoing retroperitoneal laproscopic surgery and 1 case with open surgery. In 5 laproscopic operations, the mean operative duration was 152 (120 - 235) minutes and the mean estimated intraoperative blood loss 130 (100 - 430) ml. The patient of open nephroureterectomy had an operative duration of 175 minutes and an intraoperative blood loss of 200 ml respectively. One patient underwent a local resection of ureteral carcinoma. A 81-year-old patient was diagnosed of tumor by ureteroscopic biopsy and accepted neither surgery or adjunctive therapy. All patients had a pathological diagnosis of urothelial carcinoma. One patient received local radiotherapy and chemotherapy with gemcitabine after nephroureterectomy while another one had local radiotherapy only. During a follow-up period of 6-36 months, CT showed that five cases undergoing nephroureterectomy were free of recurrence. One patient with local lymphatic metastasis had a local recurrence, developed multiple metastases at Month 3 and died of exhaustion at Month 8 post-operation. The patient with a local resection had a local bladder recurrence at Month 6 and underwent unilateral nephroureterectomy and radical cystectomy after local radiotherapy and neoadjuvant chemotherapy. Tumor progression was found in the patient without surgery. No implantation metastasis was found. CONCLUSIONS: Spontaneous urinary extravasation caused by the obstruction of primary lower ureteral cancer is rare and mostly peripelvic. Biopsy may exclude tumors for those patients with no obvious cause of ureteral obstruction. And radical operation is a preferred option.

[Diagnosis and treatment for adrenal ganglioneuroma].

OBJECTIVE: To evaluate the clinical characteristics,diagnosis and treatment for adrenal ganglioneuroma. METHODS: Thirty cases of adrenal ganglioneuroma at our hospital from 1999 to 2010 were reviewed. Their clinical, radiological, laboratory and pathologic findings as well as follow-up data were analyzed retrospectively. RESULTS: There were 12 males and 18 females with an age range of 7 - 63 (23.8 ± 11.4) years old. Tumor size ranged from 2.5 cm to 15 cm. Fourteen (46.7%) cases were found incidentally without any overt symptom. And the presenting symptoms and signs of the others were paroxysmal headache and palpitation (n = 7, 23.3%), hypertension (n = 5, 16.7%) and flank or abdominal pain (n = 4, 13.3%). All cases underwent open or laproscopic operations and their diagnosis of adrenal ganglioneuroma was confirmed pathologically. An oval or crescentic well-defined mass, low or intermediate attenuation on CT (computed tomography) scan was found in most cases. Tumors partially or completely surrounded major blood vessels in 9 cases while 7 cases had discrete punctuate calcification. The 24-hours urinary catecholamine and urinary free cortisol output were within a normal range. Seventeen cases underwent a (99)Tc-octreotide scan and negative results were found in 15 cases. Seven cases had a (131)I-Metaiodobenzylguanidine ((131)I-MIBG) scan and all results were negative. Twenty-one cases were diagnosed empirically as adrenal ganglioneuroma. One case was found pre-operatively with multiple bone metastases. Among all cases, either open operation (n = 11) or retroperitoneal laparoscopic excision (n = 19) was performed successfully. All recovered smoothly. One patient with pre-operative bone metastasis developed progression. No recurrence was detected after a follow-up period of 1 - 11 years. CONCLUSION: Mostly benign in nature, adrenal ganglioneuromas is often found incidentally without specific clinic symptoms. A relatively accurate preoperative diagnosis can be made on the basis of a detailed history and the relevant imaging and laboratory results. Surgical resection of tumor is a preferred choice with a fairy good prognosis.

Diagnosis and treatment of primary adult renal sarcoma.

OBJECTIVE: To investigate the clinical characteristics of primary adult renal sarcoma. METHODS: A total of 1654 cases with adult renal tumors were treated during 1985 to 2009 in Peking Union Medical College Hospital. Of all, 17 cases were diagnosed as primary renal sarcoma and underwent radical nephrectomy. The clinical features of 17 such patients were retrospectively analyzed. RESULTS: The first symptom of 10 (59%) cases in all renal sarcomas was abdominal mass. The pathological diagnosis was leiomyosarcoma (7 cases), rhabdomyosarcoma (2 cases), malignant fibrous histiocytoma (2 cases), low-differentiated sarcoma (2 cases), chromophobe renal cell carcinoma coexisting with liposarcoma (1 case), fibrosarcoma (1 case), embryonic sarcoma (1 case) and leiomyosarcoma (1 case). One patient died of tumor thrombus of the inferior vena cava during surgery. Finally, 15 cases were regularly followed up for 4 to 60 months. Till now, 1 had tumor-free survival for 9 months, and the other 14 cases died 2-38 months after the operation with a median survival time of 18 (range, 5-60) months. The median survival time of leiomyosarcoma group was 28 (range, 11-60) months, and 2 cases of malignant fibrous histiocytoma died 4 and 8 months after the operation respectively. CONCLUSIONS: The primary renal sarcoma has the clinical symptom similar with advanced renal cell carcinoma and has poor prognosis. Leiomyosarcoma might have relative good prognosis.

[Melanoma adrenal metastasis: report of 3 cases and literature review].

OBJECTIVE: To investigate the clinical diagnosis, treatment and prognosis of melanoma adrenal metastasis. METHODS: A total of 48 cases of malignant melanoma from 1985 to 2007 were reviewed. And three cases of melanoma adrenal metastasis were analyzed. There were 2 males and 1 female with an age of 57, 70 and 83 years old respectively. Two cases had a history of cutaneous melanoma and another one suffered previously from anal melanoma. The disease-free interval to adrenal metastasis were 25, 37, 33 months respectively. Two presented with flank or abdominal discomfort and another one had adrenal metastasis on routine examination. The maximal diameters of metastasis were 5, 8 and 12 cm respectively. The CT value of adrenal metastasis had a range of 9 - 45 Hu in plain scan. And the maximal post-contrast value was 107.5 Hu. RESULTS: One patient with concurrent lymph node metastasis received non-operative therapy with interleukin-2. The other two cases underwent complete resection of melanoma metastasis. One received adrenalectomy via retroperitoneal laparoscopic approach while another patient with tumor thrombus in inferior vena cava underwent right adrenalectomy and extraction of tumor thrombus out of inferior vena cava in traditional open surgery. When contralateral adrenal metastasis of melanoma was found 6 months later, left adrenalectomy laparoscopically was performed. The patient who was managed non-operatively died of cachexia 6 months later. Two operated patients did not accept adjuvant therapy. One had multiple metastasis in 16 months and died at 21 months. Another was alive without recurrence or new metastasis at 30 months. CONCLUSION: Melanoma metastasis to adrenal gland is rare and it generally has a poor prognosis. Patients with adrenal metastases from melanoma, either isolated or with a limited number of additional metastases, may achieve a survival benefit from surgical resection if all visible lesions are removed.

[Clinical effectiveness and safety of combined therapy with an alpha-blocker and an anticholinergic drug for patients with benign prostatic hyperplasia].

OBJECTIVE: To evaluate the effectiveness and safety of the combined therapy of doxazosin and tolterodine for BPH patients with dominant storage symptoms. METHODS: From May 2009 to April 2010, 76 patients diagnosed as benign prostatic hyperplasia (BPH) by International Prostate Symptom Score (IPSS), flowmetry, ultrasound for prostate volume, residual urine and other methods were included in the study, all of them fulfilled the study's inclusive/exclusive criteria. The principal exclusive criteria were prostate volume > 50 ml, maximum flow rate (Qmax) < 10 ml/s and residual urine > 100 ml. All 76 patients had bothering storage symptoms and divided randomly into two groups:doxazosin group (n = 36) in which patients were treated with doxazosin 2 mg once daily for 8 weeks, and combination group (n = 40) in which patients were treated with doxazosin 2 mg once daily and tolterodine 2 mg twice daily for 8 weeks. RESULTS: At baseline, there was no significant differences. The IPSS was significantly improved in the two groups after treatment, but the reduction of IPSS in combination group was significantly greater (18.7 ± 2.2 vs 12.7 ± 3.9) than that in doxazosin group (18.6 ± 3.0 vs 15.2 ± 3.8) (P < 0.05), and the decreased storage IPSS was the main contribution to the reduction of IPSS in combination group. There was no difference between the groups for Qmax and residual urine. CONCLUSIONS: Combined therapy with doxazosin plus tolterodine for BPH patients with dominant storage symptoms can improve the IPSS, especially the storage IPSS, without any negative effects on uroflow rate and residual urine.

[Study on the change of bladder wall weight pre and after surgery in patients with benign prostatic hyperplasia].

OBJECTIVE: To investigate the changes and its clinical significance of ultrasound evaluated bladder wall weight (UEBW) pre and after surgery in patients with benign prostatic hyperplasia (BPH). METHODS: The clinical data of 63 patients of benign prostatic hyperplasia and 30 of contrast control were studied. The bladder wall weight was calculated by subtracting the volume of sphere represented by the urine volume inside the bladder from the volume of the sphere representing the whole bladder (sphere with a radius = internal radius + bladder wall thickness). Bladder outlet obstruction (BOO) and bladder function was evaluated by urodynamic studies. Of the 63 patients with BPH, UEBW was (97 +/- 54) g, while the control group was only (41 +/- 14) g. UEBW was found to be significantly correlated with LinPURR grade (R = 0.47, P < 0.01) and positive residual urine volume (R = 0.48, P < 0.01), and it was negatively correlated with Qmax (R = -0.52, P < 0.01) and detrusor contraction strength (presented as WF, R = -0.4, P < 0.05). RESULTS: A significant difference was found between the UEBW pre and after surgery [(99 +/- 50) g vs. (56 +/- 21) g, t = 5.05, P < 0.01)]. UEBW decreased 43.68%, and IPSS score decreased 16.81 point, while Qmax increased 8.38 ml/s. CONCLUSION: As an non-invasive methods, measurement of bladder wall weight appears to be a useful marker in evaluating status before operation and in monitoring the effect of surgery for patients with BPH.

[Clinical effectiveness and safety study of combined therapy with an alpha-blocker and an anticholinergic for patients with LUT/BPH].

OBJECTIVES: The effectiveness and safety of the combined therapy of terazosin and tolterodine for LUT/BPH patients with dominant storage symptoms was evaluated. METHODS: The present study included 69 patients diagnosed as LUT/BPH by IPSS, flowmetry, ultrasound for prostate volume, residual urine, serum PSA and other methods, all of them fulfilled the study's inclusive/exclusive criteria. The principal exclusive criteria were prostate volume > 50 ml, Qmax < 10 ml/s and residual urine > 50 ml. All 69 patients had bothering storage symptoms after initial treatment with terazosin 2 mg once daily for one week., they were divided randomly into two groups. Terazosin group in which patients were treated with terazosin 2 mg once daily for six weeks, and combination group in which patients were treated with terazosin 2 mg once daily and tolterodine 2 mg twice daily for 6 weeks. RESULTS: Sixty-nine patients with LUT/BPH were recruited with inclusive/exclusive criteria of our study, thirty-six patients were in terazosin group and 33 were in combination group. At baseline there were no significant differences between the groups, in mean age, body weight, prostate volume, IPSS, storage IPSS, voiding IPSS, Qmax and residual urine. The results showed that the IPSS was significantly improved in the two groups after treatment, but the reduction of IPSS in combination group was significantly greater than that in terazosin group (P < 0.001), and the decreased storage IPSS was the main contribution to the reduction of IPSS in combination group. There were no differences between the groups for Qmax and residual urine. CONCLUSION: Combined therapy with terazosin plus tolterodine for LUT/BPH patients with dominant storage symptoms can improve the IPSS, especially the storage IPSS, without any negative effects on uroflow rate and residual urine. Combining with inclusion/exclusion criteria of the study, we believe that combined therapy with alpha-blocker plus anticholinergics is an effective and safe treatment for LUT/BPH patients with relatively small prostate volume, moderate impaired Q(max) and dominant storage symptoms.