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PDF file - 23K, List of Primers

PDF file - 68K, Isolation of CSCs/CICs from human RCC cells

PDF file - 74K, RT-PCR analysis

PDF file - 91K, SP analysis of DNAJB8- transduced cells

PDF file - 67K, Western blot analysis

PDF file - 142K, Protein expression of DNAJB8 in SP and MP cells

PDF file - 92K, Serine-rich region of DNAJB8 has role for induction of SP cells

This study aims to evaluate the influence of myosteatosis on survival of patients after radical cystectomy (RC) for bladder cancer. We retrospectively identified 230 patients who underwent RC for bladder cancer at our three institutions between 2009 and 2018. Digitized free-hand outlines of the left and right psoas muscles were made on axial non-contrast computed tomography images at level L3. To assess myosteatosis, average total psoas density (ATPD) in Hounsfield Units (HU) was also calculated as an average of bilateral psoas muscle density. We compared cancer-specific survival (CSS) between high ATPD and low ATPD groups and performed cox regression hazard analyses to identify the predictors of CSS. Median ATPD was 44 HU (quartile: 39–47 Hounsfield Units). Two-year CSS rate in overall patients was 76.6%. Patients with low ATPD (P = 0.01) than patients with high ATPD (≥ 44 HU). According to multivariate analysis, significant independent predictors of poor CSS were: Eastern Cooperative Oncology Group performance status ≥ 1 (P = 0.03), decreasing ATPD (P = 0.03), non-urothelial carcinoma (P = 0.01), pT ≥ 3 (P P

Our objectives were to compare measurements of ureteral wall area, ureteral wall volume and ureteral wall thickness for their use in prediction of shock wave lithotripsy outcomes. We retrospectively identified 218 patients that underwent shock wave lithotripsy for ureteral calculi with pretreatment non-contrast computed tomography. We measured ureteral wall thickness, ureteral wall area and ureteral wall volume by high functional viewer. Ureteral wall thickness was defined as the maximum thickness of ureteral wall, and ureteral wall area as the area of ureteral wall around the stone in the maximal stone diameter on axial computed tomography image. Ureteral wall volume was defined as the volume of ureteral wall from the upper to lower edge of the stone. Treatment success was defined as absence of residual fragments within 3 months after the first session. We compared the outcome predictive power among these parameters and logistic regression analysis to identify factors contributing to treatment failure. The treatment success rate was 47.6%. Ureteral wall thickness, ureteral wall area and ureteral wall volume in successful cases were all significantly smaller than those in unsuccessful cases (all p

We herein present an innovative technique of laparoscopic posterior mesh rectopexy (LPMR) for full-thickness rectal prolapse and report the clinical outcomes in our institution. Ten consecutive patients who were treated with our latest LPMR technique using mesh with an anti-adhesion coating from June 2014 to May 2017 were retrospectively analyzed. All patients were women with a mean age of 63.6 years (range, 39 to 82 y). The median operative time and blood loss volume were 197.5 minutes (range, 156 to 285 min) and 0 mL (range, 0 to 152 mL), respectively. No perioperative complications occurred, including surgical site infection, pneumonia, urinary dysfunction, and intestinal obstruction. The median follow-up duration was 768 days (range, 396 to 1150 d). During the follow-up, the cumulative incidence of full-thickness rectal prolapse and any mesh-related complications was 0. It may be possible to eliminate retroperitoneal closure using a mesh with an anti-adhesion coating. Our LPMR technique appears safe and acceptable.

Objectives The objective of the present study was to investigate the usefulness of three-dimensional images of stones to measure mean stone density for predicting the outcome of shock wave lithotripsy. Methods We retrospectively identified 239 patients who underwent shock wave lithotripsy with pretreatment non-contrast computed tomography. We automatically measured the mean stone density of three-dimensional images of stones using a high-functional viewer. For comparison, mean stone density was also measured by two previously reported techniques using both the abdominal windows and the bone windows on the axial slice at the level of the largest diameter of the stone. We compared the outcome predictive power after the first treatment with outcomes according to measurement by four other methods. We also carried out logistic regression analysis, including mean stone density measured by three-dimensional images. Results The single treatment success rate was 48.5%. The effect size (14.148) of the mean stone density measured by three-dimensional images was higher than those of the other four manual methods. In addition, the area under the curve (0.6330) of the mean stone density measured by three-dimensional images was significantly higher than those of the other methods. Increasing stone volume (P = 0.002) and increasing mean stone density measured by three-dimensional images (P = 0.023) were significant independent predictors of the treatment outcome on multivariate analysis. Conclusions This is the first study to compare the predictive powers for shock wave lithotripsy outcome of various mean stone density measuring methods. There is an indication that mean stone density automatically measured by three-dimensional images of stones is more useful than other measuring methods for predicting outcomes of shock wave lithotripsy.

In a previous study, we found that DNAJB8, a heat shock protein (HSP) 40 family member is expressed in kidney cancer stem-like cells (CSC)/cancer-initiating cells (CIC) and that it has a role in the maintenance of kidney CSC/CIC. Heat shock factor (HSF) 1 is a key transcription factor for responses to stress including heat shock, and it induces HSP family expression through activation by phosphorylation. In the present study, we therefore examined whether heat shock (HS) induces CSC/CIC. We treated the human kidney cancer cell line ACHN with HS, and found that HS increased side population (SP) cells. Western blot analysis and qRT-PCR showed that HS increased the expression of DNAJB8 and SOX2. Gene knockdown experiments using siRNAs showed that the increase in SOX2 expression and SP cell ratio depends on DNAJB8 and that the increase in DNAJB8 and SOX2 depend on HSF1. Furthermore, treatment with a mammalian target of rapamycin (mTOR) inhibitor, temsirolimus, decreased the expression of DNAJB8 and SOX2 and the ratio of SP cells. Taken together, the results indicate that heat shock induces DNAJB8 by activation of HSF1 and induces cancer stem-like cells.

Objectives To identify antigenic peptides of cancer stem-like cells (CSCs) antigen, DNAJB8, and establish a mouse CSCs-targeting immunotherapy model. Materials and methods To induce DNAJB8-specific immune reaction, we stimulated human CD8+ lymphocytes with antigen-presenting cells pulsed with a cocktail of three candidate HLA-A*24:02 restricted peptides and assessed peptide specific human cytotoxic T lymphocytes (CTLs) induction. One of the antigenic peptides showed identical amino acid sequence as corresponding mouse DNAJB8. We evaluated CTL induction with the peptide immunization in mouse model. Results We confirmed peptide-specific interferon-γ secretions and cytotoxic activities of induced human CTLs. In vivo immunization with the peptide to mice, peptide-specific CTL response could be observed in mouse CD8+ T cells. Furthermore, immunization with the peptide showed significant anti-tumor effects compared with negative controls. Conclusion DNAJB8-derived peptide is a novel candidate for CSCs-targeting immunotherapy, and mouse models can be used to evaluate CSCs-targeting immunotherapy.

Objectives To identify biomarkers predicting prognosis in bladder cancer patients undergoing the gemcitabine and cisplatin regimen. Methods We studied 52 patients with metastatic bladder cancer treated with the gemcitabine and cisplatin regimen by evaluating the relationship between the expression of two biomarkers, ribonucleotide reductase subunit M1 and excision repair cross complementing 1, by immunohistochemistry and clinical outcomes. Results The patients with low expression of ribonucleotide reductase subunit M1 showed a higher objective response rate by the gemcitabine and cisplatin regimen than those with high expression of ribonucleotide reductase subunit M1 (80.0% and 45.5%, respectively). No differences were observed according to the expression level of excision repair cross complementing 1. Low expression of ribonucleotide reductase subunit M1 significantly prolonged overall survival and progression-free survival compared with the high expression group. Low expression of excision repair cross complementing 1 tended to prolong overall survival and progression-free survival, but there were no significant differences (P = 0.07 and 0.10, respectively). Multivariate analysis showed that the expression of ribonucleotide reductase subunit M1 was the only independent prognostic factor (P = 0.012). Conclusions The expressions of ribonucleotide reductase subunit M1 seem to be associated with clinical response and survival in patients with metastatic bladder cancer treated with gemcitabine and cisplatin-based chemotherapy.

A 51-year-old man visited our hospital since he had noticed remarkable increase in size of left scrotal contents. Computed tomography revealed left testicular swelling, retroperitoneal, pelvic and left inguinal lymph nodes. Serum testicular tumor markers (α fetoprotein, β-human chorionic gonadotropin, lactate dehydrogenase) were elevated. Low left orchiectomy was performed due to swelling of the left inguinal lymph node. The excised specimen weighed 3,400g. Pathological findings were non-seminoma. Although there was no operation history of scrotal groin, there was metastasis in the left inguinal lymph node ; therefore, the stage of disease was T2N3M1a. According to the postoperative values of tumor markers, his condition was judged as "good prognosis" in the International Germ Cell Consensus Classification (IGCCC), and he underwent 3 courses of bleomycin, etoposide and cis-platin (BEP) therapy. Although the tumor markers decreased to the normal limit and the size of lymph node was remarkably decreased after 2 courses of BEP therapy, one course of EP therapy was added for further reduction of tumor size. For the residual lymph node metastasis, retroperitoneal lymph node dissection (RPLND) was performed. Then pelvic and left inguinal lymph node dissection was performed at a later date. Pathological findings of excised lymph nodes were only necrotic tissue. He is alive without disease recurrence three years after treatment.

BACKGROUND Recent large-scale randomized studies have demonstrated that 2 new hormone preparations (abiraterone and enzalutamide) prolong survival in docetaxel-treated or -naïve castration-resistant prostate cancer patients. However, no studies have directly compared antitumor effects between these 2 agents, and no clear guidelines are available for choosing between them. OBJECTIVE The objective of this clinical study is to compare antitumor effects and adverse events between abiraterone and enzalutamide by allocating castration-resistant prostate cancer patients deemed not indicated for docetaxel treatment to receive either of the 2 agents. METHODS This study is an open-label, comparative study allocating castration-resistant prostate cancer patients to abiraterone or enzalutamide treatment arms (allocation factors: age RESULTS Recruitment started in May 2016, and 13 patients have been recruited so far. We expect to complete enrollment by December 2020. CONCLUSIONS Recently, cross-resistance between abiraterone and enzalutamide has been an issue of focus. Urologists thus tend to prefer docetaxel rather than sequential therapies using 2 hormonal preparations after the progression of a first hormonal preparation. From that perspective, our clinical trial is rather out of fashion. Nevertheless, we assume that many patients receive hormonal sequential therapy in the actual clinical setting, since most such patients cannot receive chemotherapeutic agents due to old age or poor performance status. This is why we are attempting this randomized clinical trial comparing abiraterone versus enzalutamide. We will try to identify which drug is suitable for initial hormonal therapy among castration-resistant prostate cancer patients who do not meet the indications for docetaxel therapy in terms of not only antitumor effect, but also adverse events and quality of life. CLINICALTRIAL University Hospital Medical Information Network UMIN000022102; https://upload.umin.ac.jp /cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000025463 (Archived by WebCite at http://www.webcitation.org/70xaQfGlJ)

Kidney stones are a major public health concern with continuously increasing worldwide prevalence. Shock wave lithotripsy (SWL) is the first line treatment choice for upper urinary tract calculi with ureteroscopy and has advantages of safety and noninvasiveness, but the treatment success rate of SWL is lower than that of other therapies. It is therefore important to identify predictive factors for SWL outcome and select a suitable treatment choice for patients with upper urinary tract calculi. In recent years, computed tomography (CT) has become the gold standard for diagnosis of upper urinary tract calculi. Several factors based on CT images, including skin-to-stone distance, mean stone density, stone heterogeneity index, and variation coefficient of stone density, have been reported to be useful for predicting SWL outcome. In addition, a new method of analysis, CT texture analysis, is reportedly useful for predicting SWL outcomes. This review aims to summarize CT parameters for predicting the outcome of shock wave lithotripsy in stone cases in the upper urinary tract.

Background It remains controversial as to whether active stone removal should be performed in patients with poor performance status because of their short life expectancy and perioperative risks. Our objectives were to evaluate treatment outcomes of active stone removal in patients with poor performance status and to compare life prognosis with those managed conservatively. Methods We retrospectively reviewed 74 patients with Eastern Cooperative Oncology Group performance status 3 or 4 treated for upper urinary tract calculi at our four hospitals between January 2009 and March 2016. Patients were classified into either surgical treatment group or conservative management group based on the presence of active stone removal. Stone-free rate and perioperative complications in surgical treatment group were reviewed. In addition, we compared overall survival and stone-specific survival between the two groups. Cox proportional hazards analysis was performed to investigate predictors of overall survival and stone-specific survival. Results Fifty-two patients (70.3%) underwent active stone removal (surgical treatment group) by extracorporeal shock wave lithotripsy (n = 6), ureteroscopy (n = 39), percutaneous nephrolithotomy (n = 6) or nephrectomy (n = 1). The overall stone-free rate was 78.8% and perioperative complication was observed in nine patients (17.3%). Conservative treatment was undergone by 22 patients (29.7%) (conservative management group). Two-year overall survival rates in surgical treatment and conservative management groups were 88.0% and 38.4%, respectively (p

OBJECTIVES To compare various fat parameters based on computed tomography images between recurrent stone-forming patients and patients forming stones for the first time. METHODS Included in the present study were 300 patients with upper urinary tract calculi who had undergone active stone removal in our hospital. Using pretreatment computed tomography images, we measured visceral fat area and volume, subcutaneous fat area and volume, visceral fat area ratio and visceral fat volume ratio. We compared patient backgrounds and these fat parameters between those who recurrently formed stones and those who formed stones for the first time. We also performed logistic regression analysis to identify factors that contribute to severe stones. RESULTS A total of 148 (49.3%) patients were recurrent stone-forming patients. Recurrent stone-forming patients were statistically significantly younger (P < 0.01) and there were more male patients (P < 0.01). In addition, visceral fat area ratio and visceral fat volume ratio in recurrent stone-forming patients were significantly higher than those in first-time stone-forming patients (P = 0.03 and P = 0.01, respectively). On the other hand, there was no significant difference in visceral fat area (P = 0.32), subcutaneous fat area (P = 0.36), visceral fat volume (P = 0.38) or subcutaneous fat volume (P = 0.23). Receiver operating characteristics analysis showed that area under the curve of visceral fat volume ratio (0.583) for recurrent stones was larger than that of visceral fat area ratio (0.571). In multivariate analysis, increasing visceral fat volume ratio was an independent significant predictor of recurrent stones (P = 0.04). CONCLUSIONS Recurrent stone-forming patients have high visceral fat ratios compared to first-time stone-forming patients, shown here for the first time.

Although previous studies have indicated that stone heterogeneity can affect extracorporeal shockwave lithotripsy (SWL) outcomes, there is no established measurement of stone heterogeneity on CT imagery. We investigated whether variation coefficient of stone density (VCSD) can predict shockwave success.We conducted a retrospective review of 245 patients with urinary calculi who had undergone SWL. We compared the predictive powers of treatment success between VCSD and other parameters associated with CT attenuation. In addition, we performed logistic regression analysis to identify the factors contributing to treatment success. Treatment success was determined within 3 months after first treatment using noncontrast CT.The treatment success rate was 47.8% (117/245 cases). From receiver operating characteristic curves for treatment success, area under curve of VCSD (0.7181) was larger than that of mean stone density (MSD) (0.6384, p = 0.09) and standard deviation of stone density (0.5412, p 0.01). Multivariate analysis revealed that MSD (p = 0.028) and VCSD (p 0.001) independently predicted the outcome. Categorized by stone location, VCSD was the independent significant predictor for SWL outcomes in both kidney (p = 0.047) and ureteral calculi (p 0.001).We found that VCSD can be a novel predictor of SWL success. The development of nomograms or scoring systems, including VCSD, can assist in the decision process for patients and minimize unnecessary delay in treatment of urolithiasis.

A 63-year-old man was admitted to our hospital owing to weight loss and vertigo. Endoscopic examination revealed advanced gastric cancer type 2. Abdominal CT showed a 62mm liver tumor in segment 4 and a 26mm tumor in segment 8. Distal gastrectomy and D2 lymph node dissection were performed. After surgery, he was administered chemotherapy with S- 1. After 2 courses of treatment, the tumors' in segments 4 and 8 were reduced to 52mm and 16mm, respectively. No other metastases were detected. Left lobectomy and partial resection of segment 8 were performed. The pathological therapeutic effects were rated as Grade 1b for the tumor in segment 4 and Grade 3 for the tumor in segment 8. After hepatectomy, he was administered adjuvant chemotherapy with S-1 for 1 year. No recurrence has been detected for 4 years and 6months after hepatectomy.

Background: Recent large-scale randomized studies have demonstrated that 2 new hormone preparations (abiraterone and enzalutamide) prolong survival in docetaxel-treated or -naive castration-resistant prostate cancer patients. However, no studies have directly compared antitumor effects between these 2 agents, and no clear guidelines are available for choosing between them. Objective: The objective of this clinical study is to compare antitumor effects and adverse events between abiraterone and enzalutamide by allocating castration-resistant prostate cancer patients deemed not indicated for docetaxel treatment to receive either of the 2 agents. Methods: This study is an open-label, comparative study allocating castration-resistant prostate cancer patients to abiraterone or enzalutamide treatment arms (allocation factors: age

INTRODUCTION AND OBJECTIVES: Cancer stem-like cells (CSCs) are small population of cancer cells with superior tumor initating, seilf-renewel, and differentiation properties. In the previous study, we found that HSP40 family member DNAJB8 contributes to the CSC phenotype in renal cell carcinoma(RCC). HSF1 has been described to induce heat shock proteins by heat shock stress and be related to initiation and progression of cancer. In this study, we investigated the relations of HSF1 and CSC phenotype including DNAJB8. METHODS: RCC cell lines ACHN was given heat shock by incubating in 45 degrees for 60 minutes. HSF1 and DNAJB8 expression was evaluated by RT-PCR and Western blot analysis. CSCs were isolated by side population method by flowcytometry. SP(side population) cells mean CSCs, MP(main population) cells mean non-CSCs. As an inhibitor of HSF1, temsirolimus (mTOR inhibitor) was used. The cell viability was assessed by MTT assay. RESULTS: Heat shock stress enhanced the expression of DNAJB8 as well as SOX2. The expression of these molecules reached maximum 24 hours after heat shock, and the SP population increased at the same time. When MP cells sorted by flowcytometry were stimulated by heat shock, SP population could be newly detected. This fact implies that heat shock stress could induce the reversion from MP cells to SP cells. Concerning the mechanism of the reversion from MP cells to SP cells by heat shock, HSF1 seems to play an important role. We found that the expression of phosphorylated HSF1 increased by heat shock, although the expression level of HSF1 itself did not change. Interestingly, when HSF1 was knocked down by siRNA, DNAJB8 could not be induced even after heat shock stress. This suggests that the stress-inducible expression of DNAJB8 is regulated by HSF1, which bind to the heat shock element(HSE) in the promoter of DNAJB8 gene and stimulate its transcription. Treatment with temsirolimus to ACHN cells could inhibit phosphorylation of HSF1, followed by down regulation of DNAJB8, resulting in the decrease of SP population. SP cells were more susceptible by temsilolimus treatment than MP cells. CONCLUSIONS: Stress including heat shock such as hyperthermia might induce DNAJB8 expression, which is related to CSC phenotype. Phosphorylation of HSF1 is required in this process. mTOR inhibitor could inhibit CSC specifically by inhibiting phosphorylation of HSF1.