In the current study, we evaluated the expression pattern of neuroglobin (Ngb) in the astrocytes after spinal cord injury (SCI) and explore the clinical significance. For this purpose, a total of 48 Sprague-Dawley rats were divided into the SCI group (n = 40) and Sham group (n = 8). Rats in the SCI group were used to prepare the SCI models by using the modified Allen's method, followed by the HE staining to observe the post-SCI pathological changes and immunofluorescent staining to observe the dynamic changes of Ngb in astrocytes after SCI. Then, oxidative stress injury models were constructed on the astrocytes in the spinal cord of rats by using peroxide in different concentrations (0, 50, 100, 150, 200 and 400μmol/L), and at 6 and 12 h after treatment, the vitality of astrocytes that were treated by peroxide in different concentrations was determined using the MTT method, while the ability of astrocytes to generate radical oxygen species (ROS) was determined by using the flow cytometry. The mRNA expression of Ngb after the oxidative stress injury in astrocytes was measured by using the real-time quantitative PCR. Results of HE staining demonstrated that rats with SCI presented with the gradual transition from acute injury into the glial scar, a natural repair, while the results of immunofluorescent staining indicated that after SCI, expression of Ngb in the astrocytes experienced an increase followed by a decrease, and the peak level was attained at 14 d after SCI. Following the treatment of H2O2 at different concentrations (50, 100, 150, 200 and 400μmol/L) for 6 and 12 h, the vitality of astrocytes in the model groups was significantly lower than that in the control groups (all P < 0.05). As the concentration of H2O2 increased (50, 100, 150, 200, 400μmol/L) and exposure to H2O2 prolonged (6, 12 h), mRNA expression was firstly increased but then decreased in astrocytes in a time-dose dependent pattern (all P < 0.05). After SCI, the expression of Ngb in the astrocytes of spine was upregulated, suggesting that Ngb may be involved in the anti-oxidative stress injury in astrocytes after SCI, thereby playing as an endogenous protector of cells.

AIMS:This study aimed to investigate the therapeutic effect and molecular mechanism of chlorogenic acid (CGA) on cyclophosphamide (CYP)-induced rat interstitial cystitis (IC). MATERIALS AND METHODS:An animal model of IC was established by intraperitoneal injection of CYP in female Sprague-Dawley rats. Eighty rats were randomly assigned to four groups: negative control (NC), NC treated with CGA (NC + CGA), IC, and IC treated with CGA (IC + CGA). Bladder urination function was assessed by analyzing urodynamic parameters. The expression of apoptosis-related proteins and inflammatory biomarkers in bladder specimens was detected using western blot and immunohistochemistry analysis. KEY FINDINGS:Compared with the IC group, bladder urinary function was significantly improved in the IC + CGA group. CGA treatment reduced inflammatory damage in the bladder tissue of IC rats. Caspase3 and Bax expression was higher while Bcl-2 expression was lower in the IC group compared to the IC + CGA group. In addition, there were significant differences between the groups in the expression levels of inflammatory biomarkers in the bladder tissue. Furthermore, CGA could inhibit CYP-induced MAPK/NF-κB phosphorylation in the rat bladder tissue. SIGNIFICANCE:In a CYP-induced rat model of IC, CGA could reduce inflammation and apoptosis, thus partially restoring bladder function, and the MAPK/NF-κB pathway was probably involved in it.

Four most frequently mutation genes can distinct ccRCC from other tumor. UG1 and has-miR590-5p in regulating PBRM1 RNA expression contribute to the development of ccRCC.

下尿路症状(lower urinary tract symptoms,L UTS)是中老年男性患者常见的临床症状,随着我国人口老龄化步伐的加快,将有越来越多的中老年人受到LUTS的困扰.LUTS可由多种原因导致,其中最为常见的是良性前列腺增生(benign prostatic hyperplasia,BPH).而针对前列腺与LUTS的相关性,近年来的研究使我们不断加深了对其认识,本文结合2016年EAU《非神经源性男性LUTS指南》和中国泌尿外科门诊患者LUTS现状调查(LUTS ChinaI-V期)结果,对前列腺相关和不相关的中老年男性LUTS诊治现状与进展做一述评.

Interstitial cystitis (IC) is a chronic inflammation disorder mainly within the submucosal and muscular layers of the bladder. As the cause of IC remains unknown, no effective treatments are currently available. Administration of stem cell provides a potential for treatment of IC.

Background Previous studies have confirmed a family risk of nephrolithiasis (NL), but only 15% of all cases are associated with an identified monogenic factor. In clinical practice, our group encountered a patient with NL combined with cystic kidney disease that had 3 affected family members. No known mutations association with NL was detected in this family, and thus further investigation of the molecular cause of NL was deemed to be necessary. Results Quality analysis from the sequencing stage showed a more than 80-fold average depth and 95% coverage for each sample, and six mutations within six genes were chosen as candidate variants for further validation. Genotyping of rs182089527in the phosphodiesterase 1A ( PDE1A ) gene in the validation cohort indicated that the alternative allele was present in 15 patients with heterozygosity and in 1 patient with homozygosity, and exhibited significant enrichment in NL patients (Fisher’s exact test, adjusted p = 0.0042) and kidney cystic patients (Fisher’s exact test, adjusted p = 0.067) compared to controls. In addition, function analysis displayed a significant decrease in the protein and mRNA expression levels resulting from the rs182089527 mutant sequence compared with the wild-type sequence. Moreover, patients with this mutation displayed a high level of creatinine and urea in urinalysis. Conclusions Our study provides genetic evidence that the rs182089527 mutation in PDE1A is involved in the development of NL and kidney cysts, which should help to improve personalized medicine for diagnosis and treatment.

inTrodUcTion Radical cystectomy for bladder and urinary diversion is considered the gold standard for treatment of muscular invasive bladder cancer or high‐risk nonmuscular invasive bladder cancer. Urinary diversion is usually classified into three types: orthotopic neobladder, ileal conduit or sigmoid conduit, and cutaneous ureterostomy.[1] Orthotopic neobladder is a better choice for urinary diversion than the other types because of its nonurinary diversion and better quality of life when this method is feasible for patients. One of the complications after creating an orthotopic neobladder is urinary incontinence. With an increasing amount of orthotopic neobladder procedures, more patients are suffering from urinary incontinence. Therefore, an article is required for diagnosis and treatment of urinary incontinence after the orthotopic ileal neobladder procedure.

目的 探讨内质网应激在大鼠间质性膀胱炎发病过程中的作用,以及4-PBA的治疗价值.方法 45只雌性SD大鼠按随机数字表法分为3组:对照(C)组、炎症(IC)组和炎症+4-PBA(IC+4-PBA)组,每组15只.C组用生理盐水处理;IC组用鱼精蛋白联合LPS诱导;IC+ 4-PBA组由炎症模型建立时予以内质网应激抑制剂4-PBA处理.5d后应用Western blot分别测定内质网应激标记物(GRP78)、自噬流标记物(P62)、自噬标记物(LC3A/B、Beclin1)的表达量;免疫荧光检测膀胱LC3A/B的表达;HE染色检测膀胱组织病理学变化;尿动力学检测膀胱功能改变.结果 IC组GRP78、P62、LC3A/B、Beclin1表达显著高于C组(P＜0.05);4-PBA+IC组GRP78、P62、LC3A/B、Beclin1表达显著低于IC组;免疫荧光结果显示IC+ 4-PBA组LC3A/B表达显著低于IC组且高于C组(P＜0.05);HE染色结果显示IC+ 4-PBA组膀胱上皮组织完整性,炎细胞浸润和组织水肿程度都较IC组明显改善;在膀胱功能学上,IC+ 4-PBA组大鼠较IC组排尿频率显著降低,排尿间隔显著延长.结论 在鱼精蛋白联合LPS诱导的大鼠间质性膀胱炎模型中,使用4-PBA抑制膀胱内质网应激恢复自噬流有可能降低膀胱兴奋性,改善膀胱功能.

Objective To investigate the inhibitory effect on human ACHN cell line and its mice xenograft by using interferon α-1b combined with cyclooxygenase-2 inhibitor and the relevant mechanism in vitro and vivo experiment .Methods ACHN cell and the xenograft mice were devided into 4 groups(IFN-α1b,NS398,IFNα-1b+NS398 and control group).The inhibitory effects were tested by CCK8(Cell Counting Kit 8)assay after AHCN were treated for 24 h and 48 h.The expression of bcl-xl and COX-2 were detected by Western blot .The vol-ume of the xenografts of ACHN cell line and testing the expression of VEGF in xenografts were measured by immunohistochemistry assay .Re-sults Both IFNα-1b and NS398 exerted inhibitory effects on ACHN and this effects showed a rising trend with a increasing concentration of drugs.The combined group was more significant than monotherapy group (P<0.05).Western blot assay showed that IFNα-1b and NS398 downregulated the expression of bcl-xl and COX-2 in ACHN.The combined group was more significant than monotherapy group (P<0.05). The combined group has the greatest inhibitory effects on the xenografts of ACHN cell line compared with monotherapy group and control group(P<0.05).The expression of VEGF in tumor was obiviously inhibited in combined group compared with monotherapy group and con -trol group (P<0.05).Conclusion IFNα-1b combined with NS398 can inhibit the proliferation of ACHN and suppress the tumor growth .

The pathogenesis of bladder pain syndrome/interstitial cystitis (BPS/IC) is currently unclear. However, inflammation has been suggested to play an important role in BPS/IC. JNK downstream signaling plays an important role in numerous chronic inflammatory diseases. However, studies of the JNK pathway in BPS/IC are limited. In this study, we investigated the role of the JNK pathway in human BPS/IC and rat protamine sulfate (PS)-induced cystitis and examined the effect of the selective JNK inhibitor SP600125 on rat bladder cystitis. In our study, we demonstrated that the JNK signaling pathway was activated (the expression of JNK, c-Jun, p-JNK, p-c-Jun, IL-6 and TNF-α were significantly increasing in BPS/IC compared to the non-BPS/IC patients) and resulted in inflammation in human BPS/IC. Further animal models showed that the JNK pathway played an important role in the pathogenesis of cystitis. JNK inhibitors, SP600125, effectively inhibited the expression of p-JNK, p-c-Jun, IL-6 and TNF-α. The inhibition of these pathways had a protective effect on PS-induced rat cystitis by significantly decreasing histological score and mast cell count and improving bladder micturition function (micturition frequency significantly decreasing and bladder capacity significantly increasing). Therefore, JNK inhibition could be used as a potential treatment for BPS/IC.

The aim of the present study was to investigate the association between connexin (Cx)43 levels and alterations in gap junctional mediation of intercellular communication in overactive bladder syndrome (OAB), and to examine the effects of connexin inhibitor on this condition. Adult female Wistar rats with OAB following partial bladder outlet obstruction (PBBO) (OAB group, n=37) and sham-operated rats (control group, n=17) were studied. The ultrastructure of the rat detrusor was observed by transmission electron microscopy and the protein expression levels of Cx43 were analyzed using western blot analysis. Furthermore, bladder detrusor cells in both groups were cultured and cells in the OAB group were randomly divided into ten groups. In nine of these groups, 18-β glycyrrhetinic acid (18β-GA) was administered at various doses and durations. All groups were compared using fluorescence redistribution after photobleaching and a laser scanning confocal microscope. Cystometry demonstrated that gap junctions were an abundant mechanism among adjacent cells, and Cx43 protein expression levels were increased in the OAB group following 6 weeks of obstruction, as compared with the control group. Mean fluorescence recovery rates in the OAB group were significantly increased, as compared with the control group (P<0.01). Mean fluorescence recovery rates were noted following 18β-GA administration. These results suggested that upregulation of Cx43 induces structural and functional alterations in gap junctional intercellular communication following PBOO, and connexin inhibitors may be a novel therapeutic strategy for the clinical treatment of OAB.

The etiology of interstitial cystitis/painful bladder syndrome (IC/PBS) is unknown. Some unknown infectious organism may play a role. We previously detected tiny nanoparticles (NPs) in the bladder of IC/PBS. This study was to examine whether NPs could invade the superficial facet cells of the urinary mucosal epithelium and create an IC/PBS phenotype. NPs were isolated and cultured from bladder biopsies from IC/PBS patients. Eight-week-old female mice were inoculated with 0.2 ml NPs suspension directly into the bladder by transurethral catheterization. Mice were euthanized 5 weeks after instillation. Pathology, mast cells, substance P and gene expression of inflammatory mediators were measured in the bladder. Bladder ultrastructures were observed. For functional analysis, the 24-h micturition frequency and total voided urine of mice were tested. NPs instillation resulted in cystitis with increase in the ratio of bladder to body weight, histological damage, mast cell proliferation, substance P, and bladder-specific elevated gene expression of inflammatory cytokines. NPs in the bladder of nanobacteria-instilled mice were observed by transmission electron microscopy and scanning electron microscopy. Evaluation of 24-h micturition habits of NPs mice showed significantly increased micturition frequency and significantly decreased urine output per micturition when compared with control mice. Therefore, the results in the present study indicate that a systemic inoculation of NPs may have a crucial impact on inducing cystitis mimicking many of the clinical and histopathological characteristics of human IC/PBS, and NPs may be one of important etiological factors for IC/PBS.

目的 观察大鼠神经源性膀胱模型功能和组织学变化,探索尿源干细胞对大鼠神经源性膀胱的修复治疗作用.方法 45只健康清洁的正常SD大鼠按照完全随机法分为正常组、损伤组、干细胞组,每组15只.治疗组建模7d后,尾静脉注射尿源性干细胞.28 d后,尿动力学检测大鼠尿动力各项指标,肌条实验观察膀胱收缩情况,Western blot观察P2Y4在膀胱中的表达,TUNEL染色观察细胞凋亡情况.结果 干细胞组大鼠膀胱湿质量(0.31±0.01)g和膀胱湿质量/体质量值(1.19±0.41)、尿动力指标[收缩时间(112.75±16.52)s、离体膀胱逼尿肌肌条指标[收缩时间(16.47 ±2.29)s]、膀胱肌细胞凋亡率(28.22±11.08)％均优于损伤组[膀胱湿质量(0.66 ±0.07)g,膀胱湿质量/体质量值(2.30±0.26),尿动力指标:收缩时间(59.00 ±5.68)s,离体膀胱逼尿肌肌条指标:收缩时间(19.80 ±3.77)s,膀胱肌细胞凋亡率(59.80 ±14.18)％].Western blot检测结果显示干细胞组的蛋白P2Y4的表达低于损伤组.结论 尿源干细胞可促进脊髓损伤后大鼠神经源性膀胱的功能恢复.

目的 评价CX43蛋白磷酸化在大鼠逼尿肌过度活动症(detrusor overactivity,DO)中的作用,并初步探讨PP1和PP2A对CX43蛋白磷酸化修饰状态的影响.方法 建立大鼠DO模型作为实验组,假手术组作为对照组,Western blot检测CX43蛋白磷酸化水平、PP1及PP2A的表达变化,免疫荧光双染对CX43与PP1、PP2A的定位及表达变化进行检测,荧光漂白恢复实验(FRAP)比较正常组、DO组、DO+碱性磷酸酶(AP)组的细胞间通讯功能(gap junction intercellular communication,GJIC)的差异.结果 与正常组相比,DO组膀胱中的CX43蛋白的不同磷酸化修饰状态(P0、P1、P2)的表达水平均明显升高(P＜0.05),PP2A的表达明显降低(P＜0.05),PP1的表达未发现明显改变(P＞0.05).DO组的GJIC较正常组明显增加(P＜0.05),DO+ AP组的GJIC较正常组和DO组明显降低(P＜0.05).结论 CX43蛋白的磷酸化水平上调可能在DO的发病过程中发挥着重要的作用,这种磷酸化水平的上调可能与PP2A的表达减少相关.

目的 评价一种新型输尿管支架管纳米铁牵引线的抗张强度及生物相容性.方法 分别对纳米铁牵引线的抗张强度及用膀胱镜拔除普通输尿管支架管的拔管阻力进行测量并分析比较.在SD大鼠背部脊柱左侧肌肉组织中植入普通手术缝线作为对照组,在SD大鼠脊柱右侧肌肉组织中埋入纳米铁牵引线作为实验组,分别于术后1、2、4、6、8、12周取标本,HE染色,光镜观察,根据软组织植入物生物学定级评分方法对两组植入物所引起的组织学反应进行反应灶厚度定量、反应灶定性及接触面定性评分,并对得分进行对比分析.结果 纳米铁牵引线的抗张强度显著大于输尿管支架管拔管阻力(P＜0.05).术后两组植入丝线周围组织都表现出典型的异物慢性炎症反应,组织学可见两组植入丝线周围都以单核巨噬细胞系统浸润为主,并可见吞噬丝线纤维或纳米铁颗粒的异物巨细胞.生物学定级评分比较结果显示,术后1周时两组间反应灶厚度定量评分及接触面定性评分差异有统计学意义(P＜0.05),其余各时期评分两组间无明显差异(P＞0.05).结论 纳米铁牵引线具有良好的抗张强度和组织相容性.

目的 探讨神经源性膀胱患者行自我清洁导尿术对储尿功能及并发症的影响.方法 对我院72例神经源性膀胱患者进行自我清洁术.对比研究患者自我清洁导尿术前后的并发症变化情况、尿动力学改变情况以及治疗的满意情况.结果 患者在术后60 d的并发症情况较术前有显著降低(P＜0.01),其中泌尿系感染发生率由58.33％降到26.39％ (P ＜0.01),肾功能检查中尿素、肌酐、K+和尿酸较术前有较大的改善,其中尿素、肌酐和尿酸有统计学意义.患者尿动力学检测数据中,平均余尿量(PVR)、膀胱压(Pves)、膀胱容量(VH2O)均有下降,逼尿肌压力(Pdet)明显提高,括约肌协同失调(DSD)和膀胱低顺应性(BLG)比例下降(P＜0.05).在自我治疗的满意度评分中最高的为症状有所缓解,平均分为10分(总分10分);最低的为健康教育形式,仅为6.72分.结论 自我清洁导尿术能够有效地降低神经源性膀胱患者泌尿系感染的发生率,改善患者的尿动力情况,提高了患者的生存质量,有利于神经源性膀胱患者的治疗康复.

Objective To explore the feasibility of making cystoscope training model with rapid prototyping technology and its training method in cystoscope operation. Methods Constructed a cystoscope training model, and designed a set of corresponding training methods ac-cording to the process and behavioral essentials of cystoscope operation. Selected 20 students who have never learned cystoscope operation and trained them with the new designed cystoscope training mode and training methods, and the training results were verified. Results Compared before and after the training, the operation time of all the trainees were significantly shortened, and their operation steps and skills were more proficient. Conclusion Cystoscope training model can help beginners to grasp the theoretical basis of cystoscopy, operation steps, and behavioral essentials, so as to shorten the learning curve of the cystoscope operation.

Objective To determine the effects of the knockout of hyper-polarization-activated,cyclic nucleotide-gated cation channels (HCN1 channels) on the expression and function of large-conductance calcium-activated potassium channels (BK channels) in interstitial cells of Cajal (ICCs) in the mouse bladder,and investigate the significance of these effects to the excitatory regulation of bladder.Methods Forty-eight healthy clean adult wild-type C57BL/6J mice (HCN1 WT) and 48 HCN1 channels knockout C57BL/6J mice (HCN1 KO) were employed in this study.The expression of BK channels and subunits in ICCs at mRNA and protein levels was detected by RT-PCR,Q-PCR,Westem blotting and immunofluorescence staining.The inhibitor of BK channel,IBTX,was added in the isolated detrusor muscle strips to observe the effect on muscle contraction.While,intracellular calcium concentration in ICCs were measured with laser confocal microscopy after the treatment of IBTX and the agonist of BK channels,NS1619,respectively.Results Q-PCR and Western blot results indicated that the expression of 5 BK channels' subunits,α,β1,β2,β3 and β4 were significantly lower at mRNA level (P ＜0.01) and at protein level (α,β3,β4:P ＜ 0.01;β1,β2:P ＜ 0.05) in the bladder of HCN1 KO mice than HCN1 WT mice.Doublelabeling immunofluorescence staining revealed that the expression of α subunit was also lower in ICCs of HCN1 KO mice than HCN1 WT mice (P ＜ 0.05).IBTX increased the amplitudes of detrusor contraction in the detrusor strip of HCN1 KO and WT mice (P ＜ 0.01,P ＜ 0.05),but the KO mice detrusor had a lower sensitivity to IBTX (P ＜ 0.05).NS1619 decreased while IBTX increased the intracellular calcium concentration of ICCs in both WT and KO mice (P ＜ 0.05,P ＜ 0.01),but the actions of NS1619 and IBTX on KO mice were lower than WT mice (P ＜ 0.01).Conclusion Knockout of HCN1 channels decreases the expression and function of BK channels in ICCs,and it might be a compensational mechanism of the decreased contractility induced by the knockout of HCN1 channels.

目的 对比生物胶与缝扎法在肾创伤模型上的治疗效果.方法 家兔32只,均分为两组,切除右肾后分别建立左肾切割伤和部分切除伤模型,各模型均分为生物胶组和缝扎法组(n=8),记录各组出血量、止血时间,术前及术后1、2、3、4周分别检测血肌酐水平,术后l周超声检查,术后1个月取创面行病理检查.结果 肾切割伤生物胶组和缝扎组出血量分别是(3.09 ±0.56) mL和(5.77±1.33)mL,止血时间分别是(10.02±1.48)s和(66.40 ±6.35)s;肾部分切除伤两组出血量分别是(4.67±0.73)mL和(7.42±1.65)mL,止血时间分别是(16.16 ±3.51)s和(179.38±18.72)s,生物胶组与缝扎组的出血量及止血时间差异有统计学意义(P＜0.05).肾切割伤和肾部分切除伤中术后各时间段肌酐值生物胶组均明显低于缝扎组(P＜0.05).术后1个月病理检查提示生物胶组肾创面愈合较好,肾单位损伤明显轻于缝扎组.结论 生物胶相对于缝扎法在肾创伤治疗中对肾组织的损伤和肾功能的影响更小,止血效果更好,是一种更安全有效的止血方法.

Autophagy, a highly conserved homeostatic cellular process that removes and recycles damaged proteins and organelles in response to cellular stress, is believed to play a crucial role in the immune response and inflammation. The role of autophagy in bladder cystitis, however, has not well been clarified. Here we investigate the role of detrusor myocytes autophagy (DMA) in cyclophosphamide-induced cystitis animal model. 164 female Sprague-Dawley rats were randomized into three experimental groups and compared to three control groups, respectively. The expressions of microtubule-associated protein 1 light chain 3 (LC3), p-p70s6k (the phosphorylated form of ribosomal protein S6), SOD2 (superoxide dismutase 2) in the bladder muscular layer were measured using western blot. The co-location of LC3, alpha-smooth muscle actin (α-SMA), and autophagic vacuoles were investigated with double-labeled immunofluorescence and transmission electron microscopy (TEM). The expression of lL-1β, IL-6, IL-8, malondialdehyde (MDA), and glutathione (GSH) in the detrusor layer were analyzed using ELISA. The bladder inflammation and the number of mast cells in the muscular layer were analyzed by histology. The bladder function was evaluated using cystometry. In cyclophosphamide-induced cystitis, autophagy was detected in detrusor myocytes by increased LC3, p-p70s6k expression, and autophagosomes. However, the presence of enhanced inflammation and oxidative stress in the cyclophosphamide-treated group suggest autophagy of detrusor myocytes may not be sufficiently activated. Inflammation and oxidative stress were significantly decreased and the bladder histology and micturition function were significantly improved with rapamycin (RAPA, autophagy agonist) pre-treatment. In contrast, inflammation and oxidative stress were dramatically increased and the bladder histology and function were negatively affected with chloroquine (CQ, autophagy blocker) pre-treated. These findings preferentially provide evidence of the association between DMA and cyclophosphamide-induced cystitis in rats. The autophagy agonist RAPA significantly decreased the inflammation and protected the bladder function, which might be considered as a potential treatment for interstitial cystitis.