水素水はラット下部尿路閉塞モデルにおける膀胱機能改善効果を示す

　下部尿路症状（low urinary tract symptoms: LUTS）は，動脈硬化や前立腺肥大症に伴う下部尿路閉塞による膀胱血流障害が原因の一つとして考えられており，症状として排尿症状，畜尿症状，排尿後症状がある．ラットの下部尿路閉塞（bladder outlet obstruction: BOO）モデルは閉塞に伴い膀胱虚血を生じさせ，酸化ストレス状態を惹起し，LUTS を引き起こす病態モデルとして確立されている．近年，水素がもつ抗酸化，抗アポトーシス作用が注目されており，様々な臓器において組織の保護作用を示すことが解明され，様々な疾患の予防と治療に応用できることが多施設，多領域から報告されている．今回，ラットBOO モデルに対して，水素水（Hydrogen Water: H2）投与を行い，抗酸化作用の検証に加え，水素水が影響を及ぼす代表的メディエーターの探索を行った．　BOO モデルの作成には，９週齢の雌性ラットを用いた．開腹し，尿道に19G 針を沿わせた状態で尿道を結紮し，尿道の部分閉塞を作成した．作成直後からsham群，水素水非投与（BOO H（2 －））群と投与（BOO H（2 ＋））群に分け，４週間後，膀胱機能検査と組織学的，生化学的および免疫組織学的検討を行った．なお，水素水は経口投与した．　膀胱重量および一回排尿量は，sham 群に対して BOO H（2 ＋）群で有意差はなかったが，BOO H（2 －）群で有意に増加していた（重量 p0.05）．膀胱内圧測定では，BOO 群で排尿筋過活動が確認され，BOO H（2 －）群では，BOO H（2 ＋）群と比較して，排尿筋過活動回数は有意に増加していた（p<0.01）．膀胱筋層部における膠原繊維の比率については，BOO H2 （−）群でsham 群および BOO H（2 ＋）群に比して，有意に上昇していた（vs sham p<0.01, vs BOO H（2 ＋） p<0.01）．膀胱組織内8-OHdG の定量では，BOO H（2 －）群は BOO H2（＋）群に比して高値を示しており（p<0.05），8-OHdG 染色ではBOO H2 （－）群のみ筋層で著明な発現が認められた．網羅的ケモカイン/サイトカイン解析ではTNF-αが，BOO H（2 －）群においてBOO H（2 ＋）群と比較してもっとも発現が亢進しており，免疫染色では，BOO H2 （－）群で膀胱上皮粘膜下の間質および筋層においての発現の増加が認められた．一方，BOO H（2 ＋）群では，筋層部においての発現が抑制されていた．　水素水は，慢性虚血に起因する下部尿路症状の治療の一つになり得る可能性がある．本研究において，膀胱筋層の酸化ストレスを抑制することにより，膀胱筋層のTNF-α を中心とした炎症性サイトカインの抑制が，その作用機序の一つとして考えられた．　Lower urinary tract symptoms (LUTS) are considered to be caused by vascular insufficiency to bladder, and include voiding, urine collection, and post-voiding symptoms. The bladder outlet obstruction (BOO) model of rat is used as a pathological model, in which ischemia of the bowel is induced by obstruction, followed by chronic ischemia of the bowel and oxidant stress, resulting in LUTS. Since the antioxidant and anti-apoptotic actions of hydrogen have been attracting attention in recent years, it has been clarified that hydrogen could protect various organs, and multiple facilities and fields have reported that hydrogen could be used in the prevention and treatment of various diseases. In this study, we administered hydrogen water (H2) to the BOO model rats to investigate its effects and mechanism of action.　We used 9-week old female rats to prepare the BOO model. After laparotomy, a 19G needle was placed along the urethra and sutured to the urethra to induce partial obstruction of the urethra. Immediately after the preparation, the rats were divided into the sham, hydrogen water no-administration (BOO H2(－)), and administration (BOO H2(＋)) groups to compare the effects of intravesical pressure, and the histological, biochemical and immunohistological changes in 4 weeks.　The weight of the urinary bladder increased significantly in the BOO groups, 3.0- and 2.3- fold in the BOO H2(－) and BOO H2(＋) groups, respectively, compared with the sham group. The urination volume in one voiding increased significantly in the BOO groups, compared with the sham group. In the measurement of intravesical pressure, hyperreflexia of the urinary bladder was confirmed in the BOO groups, and the degree of detrusor overactivity was increase significantly in the BOO H2(－) group, compared with the BOO H2(＋) group. The proportion of collagen fiber in the bladder musculature increased significantly in the BOO H2(－) group, compared with the sham and BOO H2(＋) groups. In the quantification of 8-OHdG in the bladder tissues, the levels in the BOO H2(－) group were higher than the BOO H2(＋) group. In the 8-OHdG staining, significant expression was observed in the muscle layers only in the BOO H2(－) group. In the exhaustive chemokine/cytokine analysis, TNF-α was obviously inhibited in the comparison between the BOO H2(－) and BOO H2(＋) groups. In fact, TNF-α immune tissue imaging, expression was increased in the interstitium beneath the bladder cell mucosa and the muscle layers of the BOO H2(－) group. In the BOO H2(＋) group, TNF-α expression was reduced in the muscle layers.　Hydrogen water would be useful treatment as new alternative therapy for LUTS caused by the ischemic change of bladder. In this study, one of the mechanism of action of hydrogen water should be inhibition of various cytokines with a focus on TNF-α via inhibition of the oxidant stress

Pathological analysis of spermatic dysfunction following testicular ischemia-reperfusion injury\n

　Introduction & Objectives: Torsion, which may result in testicular ischemia, requires emergency surgery to restore testicular blood flow. However, the risk of spermatic dysfunction remains even if surgery is performed. The pathology of spermatic dysfunction in testicular ischemia-reperfusion injury (TIRI) remains unclear. A previous study showed the relevance of inflammation and oxidative stress in the other organs of ischemia-reperfusion injury. We hypothesized that inflammation and oxidative stress play key roles in causing spermatic dysfunction following TIRI. We investigated the pathophysiology of spermatic dysfunction in TIRI focusing on inflammatory changes using TIRI model mice.　Materials and Methods: The study used C57BL/6J male mice aged 10 to 15 weeks. To create TIRI model mice, the unilateral (left side) testicular vessels were clamped using Dieffenbach clamps (Bulldog clamps) for 1 hour and de-clamped. The bilateral testes were removed at 0 (ischemic state), 1, 3, and 5 weeks after creating the TIRI model mice. Spermatic changes following TIRI were investigated by analyzing the histology of the testes and semen and assessing levels of inflammation and oxidative stress. Semen was collected from the bilateral cauda epididymites and investigated using the sperm motility analysis system (SMAS).　Results: Histological analysis after hematoxylin-eosin staining showed tissue thickening in interstitial tissues at week 1 and 3 on the left (affected) testis, and week 1, 3 and 5 on the right (unaffected) testis. The infiltration of lymphocytes-predominant inflammatory cells were observed at week 1 and week 3 on the left (affected) testis. The destruction of ductal structures and giant cells were observed at weeks 3 and 5 on the left (affected) testis and week 5 on the right (unaffected) testis. SMAS showed significantly decreased spermatic concentration and motility in both testes of TIRI model mice compared with those of sham-operated mice at weeks 1, 3 and 5. Inflammation analysis using an inflammation-related proteome assay showed significantly increased levels of cytokines (IL-2, IL-3, IL-17A, and IL-23) and chemokines (CCL2, CCL5, CXCL1, and CX3CL1) at weeks 1, 3, and 5 in both testes of TIRI model mice. For the assessment of oxidative stress, enzyme-linked immuno-sorbent assay (ELISA) for 8-hydroxy-2’-deoxyguanosine (8-OHdG) was performed, which showed that levels of 8-OHdG were significantly increased in the left (affected) testis of TIRI model mice compared with that of sham-operated mice at all observation periods. Meanwhile, ELISA showed that levels of 8-OHdG in the right (unaffected) testis were significantly increased in TIRI model mice at weeks 3 and 5 compared with that of sham-operated mice.　Conclusions: Spermatic dysfunction following TIRI is induced by inflammation and oxidative stress. Inflammation and oxidative stress may be novel regulatory factors to prevent spermatic dysfunction following TIRI

Pathophysiological analysis of detrusor overactivity following partial bladder outlet obstruction

　Introduction: Detrusor overactivity (DO) following partial bladder outlet obstruction (PBOO) is a common urological condition in humans, with 50-70% patients with PBOO complicated with DO. The pathological mechanisms of DO following PBOO are largely unknown, but inflammatory changes may play a key role. We hypothesized that inflammation is important in the earlier pathophysiological phase before overproduction of oxidative stress in DO following PBOO. Therefore, we investigated the relationships among bladder function, ischemia, oxidative stress and inflammation in DO following PBOO in PBOO model mice.　Materials and Methods: C57BL/6J male mice aged 10 to 15 weeks were used in the study. PBOO model mice were created surgically by ligation of the proximal urethra with 5-0 nylon suture under inhalation anesthesia. Sham-operated mice were used as controls. Pathophysiological changes in the bladder at 1, 3 and 5 weeks after creation of the PBOO model mice were compared with those in sham-operated mice using functional, histological, biochemical and immunohistochemical analyses.　Results: Functional analysis using a pressure flow study showed increased maximum detrusor pressure at 1 week and DO from 3 to 5 weeks after creation of the PBOO model. Histological analysis using hematoxylin-eosin and Masson-Trichrome staining showed greater invasion of inflammatory cells and fibrosis in PBOO model mice compared with sham-operated mice at 3 and 5 weeks. Inflammatory cells were mainly present in interstitial tissue, and fibrosis gradually infiltrated from interstitial tissue to the muscular layer. Ischemia analysis showed significantlyincreased HIF-1α in PBOO model mice at all time points. Oxidative stress analysis indicated significantly increased levels of ROS from 1 week and 8-OHdG from 3weeks in PBOO model mice. An inflammation-related proteome assay showed high levels of colony stimulating factor (CSF) family proteins at 1 week and IL-2, IL-3, IL-17A, IL-23, MMP-3, MMP-9 and periostin from 3 to 5 weeks in PBOO model mice.　Conclusions: Oxidative stress and inflammatory changes showed contemporaneous increase in pathophysiology of detrusor overactivity following partial bladder outlet obstruction. Especially, CSF family and ROS changes are showed in the early stage, and might be a predict marker in the pathophysiology of DO following PBOO at the early stage

Preventive effect of indoleamine 2,3-dioxygenase 1 inhibition on lipopolysaccharide-induced prostatitis

　Introduction and Objectives: Bacterial infections are the main cause of acute prostatitis and are treated with appropriate antimicrobial therapy. However, approximately 5% of patients continue to have inflammatory symptoms even after receiving antibacterial therapy, leading to refractory conditions. Bacterial prostatitis requires additional therapy, focusing on inflammatory changes. Indoleamine 2,3-dioxygenase 1 (IDO1) catalysis is the first rate-limiting step of tryptophan metabolism. IDO1 is expressed in the prostate and plays a key role in the immune response. As the first step in investigating the relationship between acute prostatitis and IDO1, we investigated the preventive effect of IDO1 inhibition on lipopolysaccharide (LPS)- induced prostatitis using IDO knockout (Ido1 −/−) mice in this study.　Materials and Methods: The study used Ido1 −/− and wild-type (Ido1 +/+) C57BL/6J malemice aged 10–15 weeks. LPS Escherichia coli O26 (100μg/PBS, 100μL) was administered transurethrally into the lower urinary tract to create a mouse model of LPS-induced prostatitis. The prostates were removed 1, 3, 5, and 7 days after creating the model mice. Histological, immunohistochemical, and biochemical analyses were used to compare the preventive effect in Ido1 −/− mice compared with that in Ido1+/+ mice.　Results: HE staining showed suppression of ductal destruction following infiltration of inflammatory cells in Ido1 −/− mice compared with Ido1 +/+ mice. The enzyme-linked immunosorbent assay (ELISA) method was used for kynurenine pathway analysis, which showed significantly maintained tryptophan levels and decreased L-kynurenine levels in Ido1 −/− mice compared to Ido1 +/+ mice. The IDO1 assay in Ido1 +/+ mice showed significantly increased levels during all observation periods after creating the model compared with that under normal conditions. Immunofluorescent staining using five types of cytokines and chemokines (IL-2, IL-4, IL-17, CCL2, and CCL3) related to the pathophysiology of acute prostatitis showed decreased expression of these cytokines and chemokines in Ido1 -/- mice compared with Ido1 +/+ mice. Inflammation-related proteome assays showed decreased levels of IL-1β, IL-4, IL-5, IL-6, IL-17, CCL2, CCL3, CXCL1, CXCL11, and tissue inhibitor of matrix metalloproteinases (TIMP)-1 in Ido1 −/− mice compared with Ido1 −/− mice during all observation periods after model creation.　Conclusions: IDO1 is involved in LPS-induced prostatitis through cytokines and chemokines. IDO1 inhibition contributes to the prevention of LPS-induced prostatitis. IDO1 inhibition has the potential to serve as an additional therapy for acute prostatitis

Treatment outcomes of laparoscopic radical prostatectomy at Kawasaki Medical School Hospital

　Laparoscopic radical prostatectomy (LRP) was carried out in 196 patients with prostate cancer between December 2009 and November 2017 at Kawasaki Medical School Hospital, and the therapeutic outcomes were assessed. An extraperitoneal approach was used in all cases except 1 and the median follow-up period was 55 months (range, 10-117 months). The median patient age was 69 years (range, 56-79 years), median body mass index was 23.3 kg/m2 (range, 15.2-33.2 kg/m2 ), and median prostate-specific antigen (PSA) level at diagnosis was 7.4 ng/mL (range, 2.2-42.0 ng/mL). Clinical stages of T1c, T2a, T2b, T2c, T3a, and T3b accounted for 63, 43, 31, 57, 1, and 1 case, respectively, while Gleason scores at biopsy of ≥ 6, 7, and ≥ 8 accounted for 26, 138, and 32 cases, respectively. The median prostate volume was 22.0 mL (range, 7.3-65.6 mL), median operating time was 266 minutes (range, 142-540 minutes), and median blood loss (including in urine) was 650 mL (range, 10-5,800 mL). During the initial induction period, 94 patients received autologous blood transfusion and 7 received allogeneic blood transfusion. Nerve-sparing prostatectomy was performed in 17 cases (bilateral in 3, unilateral in 14). Capsular invasion was observed in 57 cases (29.1%) and positive resection margins were observed in 51 cases (26.4%). The median indwelling catheter duration was 6 days (range, 4-26 days) and the median hospital stay after surgery was 11 days (range, 8-34 days). The main complications were intraoperative rectal injury in 7 cases (3.6%), postoperative inguinal hernia in 28 (14.3%), and urethral stenosis in 8 (4.1%). The rate of urinary incontinence at ≥ 1 year after surgery was 32.7% and the rate of PSA recurrence was 15.8%. The overall survival rate was 95.6% at 5 years and 94.7% at 10 years. In conclusion, the oncological outcomes were similar to that reported by previous reports, but postoperative stress urinary incontinence and complications were slightly worse. In the future, further improvement of the surgical technique was desired

Outcomes of robot-assisted partial nephrectomy in the treatment of renal cell carcinoma at Kawasaki Medical School Hospital

　Robot-assisted partial nephrectomy (RAPN) was introduced in our hospital for treating small renal cell carcinoma in May 2018; we examined treatment outcomes in 24 patients (25 kidneys) who had undergone this procedure till 2019. The median observation period was 11 months (range, 1-17 months). The patients’ age range was 43-77 years (median, 68 years). Fourteen men and 10 women underwent the procedure. Their BMI was 17.9-39.7 (median, 24.1) kg/m2 . In one patient, RAPN was performed twice at different times for treating bilateral renal cancer. The right kidney was affected in 12 cases and the left kidney in 13 cases. The clinical cancer stage was T1a in 20 cases and T1b in 5 cases. Tumor sizes were 0.9-6.2 cm (median, 2.5 cm), and RENAL nephrometry scores were 4-10 (median, 7). The transperitoneal approach was used in 22 cases, and the retroperitoneal approach in 3. The operating durations were 147-358 min (median, 225 min), console durations were 59-394 min (median, 152 min), and renal ischemia durations were 8-54 min (median, 21 min). Blood loss was 10-700 ml (median 10 ml), and none of the patients underwent blood transfusion. The histopathological analysis of the resected tumors revealed clear cell renal cell carcinoma in 20 cases, chromophobe renal cell carcinoma in 2 cases, and papillary renal cell carcinoma, angiomyolipoma, and leiomyoma in 1 case each. All margins were negative. The postoperative hospital stay lengths were 5-14 days (median, 9 days). The postoperative deterioration in renal function was mild, and there were no severe complications. In the early stages after its introduction, RAPN was safely performed and allowed for the preservation of renal function. We plan to continue studying more cases going forward

当院の女性過活動膀胱患者における抗コリン薬とβ3アドレナリン受容体作動薬の内服継続率に関する検討

　過活動膀胱（overactive bladder：以下OAB）は，尿意切迫感や頻尿などの下部尿路症状を有し，加齢とともに増加する傾向がある．OAB 患者の治療薬として，本邦では抗コリン薬とβ3アドレナリン受容体作動薬が推奨，使用されている．今回，女性OAB 患者に対する，抗コリン薬とβ3アドレナリン受容体作動薬の内服継続率，副作用，内服中止理由に関してretrospective に検討した．対象は，2013年１月から12月の１年間に当院泌尿器科を受診した初診の女性OAB 患者，87名とした．β3アドレナリン受容体作動薬投与群と抗コリン薬投与群の内服継続率は，β3アドレナリン受容体作動薬投与群は，12か月で38.8％，60か月で18.1％，抗コリン薬投与群は，12か月18.4%，60か月で7.9％であり，β3アドレナリン受容体作動薬投与群の方が若干継続率は良いものの，両群間に差は認めなかった．副作用に関しては，抗コリン薬投与群の方が多く，口喝が17例（44.7%），便秘が15例（39.5%）であった．薬剤中止の理由は両群とも自然寛解によるものが多かった．　Overactive bladder (OAB) involves lower urinary tract symptoms such as urinary urgency and polyuria, and tends to increase with age. In Japan, the drugs recommended and used for treatment of OAB patients are anticholinergic agents and β3 adrenalin receptor agonists. The present study was a retrospective investigation of the rates of long-term administration of anticholinergic agents and β3 adrenalin receptor agonists, adverse effects with these drugs, and reasons for discontinuation of administration, with female OAB patients. The subjects were 87 female patients who were examined at this hospital\u27s Urology Dept. over 1 year between January and December 2013, and diagnosed as having OAB for the first time. With respect to the rates of long-term administration, in the β3 adrenalin receptor agonist group the rates of administration for 12 and 60 months, respectively, were 38.8% and 18.1%, and these rates in the anticholinergic agent group were 18.4% and 7.9%, so the long-term administration rates were somewhat higher in the β3 adrenalin receptor agonist group, but no difference between the groups was found. Adverse effects were more frequent in the anticholinergic agent group, with 17 subjects in that group (44.7%) developing buccal dryness, and 15 (39.5%) developing constipation. In both groups, the most frequent reason for discontinuation of administration was spontaneous remission