Homology of lungs and gas bladders: Insights from arterial vasculature

Gas bladders of ray‐finned fishes serve a variety of vital functions and are thus an important novelty of most living vertebrates. The gas bladder has long been regarded as an evolutionary modification of lungs. Critical evidence for this hypothesized homology is whether pulmonary arteries supply the gas bladder as well as the lungs. Pulmonary arteries, paired branches of the fourth efferent branchial arteries, deliver blood to the lungs in osteichthyans with functional lungs (lungfishes, tetrapods, and the ray‐finned polypterid fishes). The fact that pulmonary arteries also supply the respiratory gas bladder of Amia calva (bowfin) has been used to support the homology of lungs and gas bladders, collectively termed air‐filled organs (AO). However, the homology of pulmonary arteries in bowfin and lunged osteichthyans has been uncertain, given the apparent lack of pulmonary arteries in critical taxa. To re‐evaluate the homology of pulmonary arteries in bowfin and lunged osteichthyans, we studied, using micro‐CT technology, the arterial vasculature of Protopterus, Polypterus, Acipenser, Polyodon, Amia, and Lepisosteus, and analyzed these data using a phylogenetic approach. Our data reveal that Acipenser and Polyodon have paired posterior branches of the fourth efferent branchial arteries, which are thus similar in origin to pulmonary arteries. We hypothesize that these arteries are vestigial pulmonary arteries that have been coopted for new functions due to the dorsal shift of the AO and/or the loss of respiration in these taxa. Ancestral state reconstructions support pulmonary arteries as a synapomorphy of the Osteichthyes, provide the first concrete evidence for the retention of pulmonary arteries in Amia, and support thehomology of lungs and gas bladders due to a shared vascular supply. Finally, we use ancestral state reconstructions to show that arterial AO supplies from the celiacomesenteric artery or dorsal aorta appear to be convergent between teleosts and nonteleost actinopterygians. J. Morphol., 2013. © 2013 Wiley Periodicals, Inc.     

TUR-BT术后膀胱灌注A群链球菌联合丝裂霉素治疗高危非肌层浸润性膀胱癌的疗效观察

目的：总结经尿道膀胱肿瘤电切（TUR-BT）术后辅以沙培林（注射用A群链球菌）联合丝裂霉素（MMC）膀胱内灌注治疗高危膀胱癌的疗效。方法：回顾性研究2009年1月~2012年8月我院收治的符合高危非肌层浸润性膀胱癌患64例,观察组：TUR-BT术后行沙培林联合MMC膀胱内灌注化疗32例,对照组;MMC单药灌注32例。结果：观察组患者,平均年龄63.7岁,治疗随访时间为6~54个月,中位时间27.3个月。治疗随访期间有3例患者出现膀胱内肿瘤复发（9.3%）,1例患者疾病进展,发展为肌层浸润性膀胱癌,于术后7个月死亡（3.1%）。与对照组患者相比,疾病复发率及进展率均明显改善。结论：高危非肌层浸润膀胱癌临床复发率、进展率高,TUR-BT术后沙培林联合MMC膀胱内灌注通过局部化疗及免疫治疗联合,可有效控制疾病的复发和进展,降低患者接受膀胱部分切除或膀胱全切手术的机率,值得推广。     

Growth and characterization of multicellular tumor spheroids of human bladder carcinoma origin

Summary We have examined the MGH-U1 human bladder carcinoma cell line and 12 primary bladder carcinoma biopsies for their ability to form spheroids in suspension culture and in multiwell dishes. MGH-U1 cells formed tightly packed spheroids with a necrotic center and viable rim whereas three sublines formed loose aggregates only. Spheroids formed from as few as 100 MGU-U1 cells placed into multiwells. MGH-U1 cells derived from spheroids formed new spheroids more rapidly and consistently than cells derived from monolayer culture. Spheroid diameter increased at a rapid rate of ∼100 μm/d in multiwell dishes, and necrosis occurred only in spheroids of diameter >1 mm. Spheroids placed in spinner culture at a higher concentration (∼1.5 spheroids/ml) grew more slowly and developed necrosis at smaller diameters. The width of the viable rim of spheroids grown in spinner culture was maintained at ∼190 μm over a wide range of spheroid diameters (400 to 1000 μm). Sequential trypsinization of spheroids, which stripped layers of cells from the spheroids, demonstrated no difference in the plating efficiency of cells derived from varying depths into the spheroid. Only one of the 12 primary bladder biopsy specimens demonstrated an ability to form spheroids. This biopsy, designated HB-10, formed spheroids that grew linearly over 40 d, formed colonies in methylcellulose culture and grew as xenografts in immune-deprived mice. These studies characterize the MGH-U1 spheroids that are useful in vitro models to study the effects of various treatments for solid tumors and demonstrate the limited capacity of cells from primary human bladder biopsies to form spheroids. Supported in part by a grant from the National Cancer Institute of Canada and by grant CA29526 NCI through the National Bladder Cancer Project, U.S.A.     

Chloride-bicarbonate exchange in the urinary bladder of the turtle. Independence from sodium ion

The rates of Cl^? absorption and HCO^?₃ secretion were not different in turtle urinary bladders bathed in Na⁺-containing and solutions.These results in turtle bladder are inconsistent with Na⁺-anion cotransport but can be accounted for by a Cl^?/HCO^?₃ exchange system.     

TURBT术后即刻膀胱灌注不同浓度吡柔比星对浅表性膀胱癌患者免疫功能及生活质量的影响

目的:研究经尿道膀胱肿瘤电切术(TURBT)术后即刻膀胱灌注不同浓度吡柔比星对浅表性膀胱癌(SBC)患者免疫功能及生活质量的影响。方法:选择从2015年6月到2018年6月在我院治疗的SBC患者126例作为此次研究对象。依据随机数字法将患者分成观察组以及对照组,每组各63例,两组患者均常规给予TURBT治疗,手术完成后即刻,观察组患者采用30 mg的吡柔比星~+30 m L浓度为5%的葡萄糖液行胱灌注化疗,对照组采用30 mg的吡柔比星~+50 m L浓度为5%的葡萄糖液行胱灌注化疗。治疗1年后对比两组复发情况、免疫功能指标、生活质量以及不良反应。结果:观察组的复发率明显低于对照组,且复发时间明显长于对照组,差异均有统计学意义(P0.05)。两组治疗后的CD3~+、CD4~+、CD8~+水平及生活质量各方面评分均明显高于治疗前,差异有统计学意义(P0.05);两组治疗前及治疗后的CD3~+、CD4~+、CD8~+水平及生活质量各方面评分相比差异无统计学意义(P0.05)。观察组各种不良反应及其总发生率与对照组相比差异无统计学意义(P0.05)。结论:SBC患者在TURBT术后即刻实施吡柔比星膀胱灌注能够提升其免疫功能及生活质量,但30 mg的吡柔比星~+30 m L浓度为5%的葡萄糖液的膀胱灌注用药方案能够获得更低的术后复发率,复发时间也随之延长。     

Characterization of a fibroblast cell from the urinary bladder wall

Summary For the first time we report on the growth, culture, and matrix production characteristics of a cell type isolated from the lamina propria of the urinary bladder wall. A fibroblastlike cell was identified as distinct from bladder detrusor smooth muscle cells and urothelium based on morphology, growth characteristics, and immunohistochemical staining. Characterization of extracellular matrix synthesis by this cell type using³⁵S-methionine metabolic labeling demonstrated that these cells are capable of secreting components of the surrounding connective tissue, including several fibrillar collagens, a basement membrane collagen, and fibronectin.     

膀胱子宫内膜异位症的超声诊断(附三例临床分析及文献复习)

本文通过3例经活检和手术证实的膀胱子宫内膜异位症的超声图像分析,讨论了膀恍子宫内膜异位症的超声诊断价值。 半数以上的膀胱子宫内膜异位症有盆腔手术史。临床表现为月经期伴尿频尿急和尿痛等尿道刺激的症状,抗炎治疗常难奏效,而月经净后症状自行缓解和消退。 超声图见子宫峡部的前壁外缘有不规则的隆起,呈低回声区并向膀胱后壁突起。在月经来潮前和来潮时此隆起包块稍见增大饱满。用丹那唑或三苯氧胺治疗病灶将逐渐缩小。本文比较了膀胱镜检查的优越性及其局限性,认为超声检查可作为首选的检查方法进行早期诊断,尤其对需采取保守治疗的患者,超声检查可作定期随访以观疗效,而不需反复进行膀胱镜检。超声检查此症应与膀胱血管瘤,膀胱癌,息肉和炎症溃疡等鉴别。     

膀胱功能代偿系统的研究

本文对截瘫病人膀胱功能障碍的重建机理和方法进行了综合分析,对临床上解决膀胱功能代偿的各种途径加以比较,提出膀胱功能代偿系统-膀胱控制器的新设计,它将会使我国广大的膀胱功能障碍患者受益。     

Urinary cytology and the Paris system for reporting urinary cytology: Implications for urological management

By reducing the rate of indeterminate (atypical) diagnoses and standardising reporting terminology, The Paris System for Reporting Urine Cytology helps focus the application of cytology towards the detection primarily of high‐grade urothelial carcinoma. We present a urology‐based perspective of how the new system has influenced clinical decision‐making.     

Bladder cancer diagnosis from bladder wash by Fourier transform infrared spectroscopy as a novel test for tumor recurrence

This study proposes Fourier Transform Infrared (FTIR) spectroscopy as a more sensitive, rapid, non‐destructive and operator‐independent analytical diagnostic method for bladder cancer recurrence from bladder wash than other routinely used urine cytology and cystoscopy methods. A total of 136 patients were recruited. FTIR spectroscopic experiments were carried out as a blind study, the classification results of which were then compared with those of cytology and cystoscopy. Firstly, 71 samples (n = 37; bladder cancer and n = 34; control) were studied with transmittance FTIR spectroscopy. After achieving successful differentiation of the groups, to develop a more rapid diagnostic tool and check the reproducibility of the results, the work was continued with different samples (n = 65 as n = 44; bladder cancer and n = 21; control), using the reflection mode (ATR) of FTIR spectroscopy by a different operator. The results revealed significant alterations in moleculer content in the cancer group. Based on the spectral differences, using transmittance FTIR spectroscopy coupled with chemometrics, the diseased group was successfully differentiated from the control. When only carcinoma group was taken into consideration a sensitivity value of 100% was achieved. Similar results were also obtained by ATR‐FTIR spectroscopy. This study shows the power of infrared spectroscopy in the diagnosis of bladder cancer.

     

Effect of DHLA on response of isolated rat urinary bladder to repetitive field stimulation

Lipoic acid is an essential coenzyme in the oxidation of pyruvate and -ketoglutarate. It is easily converted to its reduced form, dihydrolipoic acid (DHLA), in vivo thereby forming a redox pair. DHLA is important in the maintenance and integrity of specific neuronal and subcellular membranes. In the present study we investigated the effect of DHLA on the response of isolated rat bladder strips to repetitive field stimulation (FS), a method used to exhaust synaptic stores of acetylcholine resulting in nerve and synaptic damage.Isolated strips of rat urinary bladders were separated into 4 groups. Group 1 strips were incubated with choline + acetyl-CoA; Group 2 strips with choline, acetyl-CoA + DHLA; and Group 3 with DHLA. Group 4 strips were controls. All strips in Groups 1–3 were subjected to 2 h of repetitive FS followed by 2 h of recovery.DHLA had no effect on the progressive decrease in contractile response observed during repetitive stimulation. However, strips incubated in the presence of DHLA showed a significantly greater degree of recovery than strips incubated in the absence of DHLA. We believe that the protection of the contractile response is related to DHLA's ability to protect nerve and/or muscle membranes from oxidative damage.     

THE DISTINCT STEPS OF CELL DETACHMENT DURING DEVELOPMENT OF MOUSE UROEPITHELIAL CELLS IN THE BLADDER

In developing mouse urinary bladder the urothelial cells respond to urine accumulation by cell detachment, i.e. desquamation. To elucidate the steps of urothelial cell detachment during embryonic development and first urine accumulation in bladder lumen, superficial and intermediate cell layers were investigated. Different electron microscopic and cytochemical methods have been used. It was possible to distinguish between an early and late events on the basis of morphology. At the beginning cell detachment involves interruption of tight junctions between neighbouring superficial cells and the formation of numerous endocytotic vesicles. Endocytotic and cup-shaped vesicles then fuse and form large parallel rows of vacuoles and multivesicular bodies in desquamating superficial cells and in the neighbouring superficial and intermediate cells. These observations clearly demonstrate the existence of specific steps during the detachment of embryonic uroepithelial cells. Apoptosis accompanies cell desquamation. Only predetermined superficial bridge cells die in an apoptotic manner.     

Pro-apoptotic effect of a nitric oxide-donating NSAID,NCX 4040, on bladder carcinoma cells

Nitric oxide-releasing non steroidal anti-inflammatory drugs (NO-NSAIDs) are a promising class of compounds that cause cell cycle perturbations and induce apoptosis in cell lines from different tumors. We investigated the activity of a recently developed NO-NSAID (NCX 4040) in bladder cancer cell lines (HT1376 and MCR). Cells were treated with different drug concentrations for different exposure times. Cytostatic and cytocidal activity was tested by SRB assay and apoptosis was evaluated by TUNEL analysis, ANNEXIN V assay and fluorescence microscopy. To further investigate the cell death-inducing mechanisms of NCX 4040, we analyzed gp-170, caspase expression and mitochondrial membrane potential (Δ Ψ) depolarization. NCX 4040 showed a striking cytocidal activity in both cell lines, reaching LC₅₀ at a 10-μ M and 50-μ M concentrations in HT1376 and in MCR cells, respectively, after an exposure of only 6 h followed by an 18-h washout. Apoptosis was triggered in up to 90% of cells and was associated with active caspase-3 expression and Δ Ψ depolarization in both cell lines after a 6-h exposure. In conclusion, NCX 4040, which probably causes apoptosis via a mitochondrial-dependent mechanism, could prove to be a useful agent for improving bladder cancer treatment.This study was supported by Istituto Oncologico Romagnolo, Forlì, and by the Italian Ministry of Health, 2002.     

Discovery of urine biomarkers for bladder cancer via global metabolomics

Bladder cancer (BC) is latent in its early stage and lethal in its late stage. Therefore, early diagnosis and intervention are essential for successful BC treatment. Considering the limitations of current diagnostic tools, noninvasive biomarkers that are both highly sensitive and specific are needed to improve the overall survival and quality of life of patients. With the advent of systems biology, “-omics” technologies have been developed over the past few decades. As a promising member, global metabolomics has increasingly been found to have clear potential for biomarker discovery. However, urinary metabolomics studies related to BC have lagged behind those of other urinary cancers, and major findings have not been systematically reported. The objective of this review is to comprehensively list the currently identified potential urinary metabolite biomarkers for BC.     

Characterization of upper lamina propria interstitial cells in bladders from patients with neurogenic detrusor overactivity and bladder pain syndrome

The upper lamina propria (ULP) area of interstitial cells (IC) in bladder has been studied for more than a decade in several species including human beings. Nevertheless there is still lack of uniformity in terminology of this cell layer. The aim of the present study was to add new data to the morphological and immunohistochemical phenotype of these cells and to find out whether this phenotype is changed in bladders from patients with neurogenic detrusor overactivity (NDO) and bladder pain syndrome (BPS). Bladder tissue was obtained from a control group and from patients with NDO and BPS. Samples were processed for morphology, electron microscopy and immunohistochemistry. A morphological and immunohistochemical phenotype for the ULP IC was assessed and changes in this phenotype were looked for in samples from patients with NDO and BPS. The ULP IC were characterized ultrastructurally by the presence of actin filaments with densifications, many caveolae and abundant rough endoplasmic reticulum (RER); on immunohistochemistry ULP IC were immunoreactive for α-sma, vimentin, CD10 and podoplanin and categorized as interstitial Cajal-like cells (ICLC). In NDO and BPS bladders we found a phenotypical shift towards a fibroblastic phenotype which was even more pronounced in the NDO group. In both groups there was also an increased presence in ULP lymphocytes. The ULP area in the human bladder contains a population of ICLC with distinct ultrastructural morphology and immunohistochemical phenotype. Their unique α-sma(+) /desmin(-) /CD34(-) phenotype allows studying this population in various bladder disorders. In bladders form patients with BPS and NDO, we observed these ULP ICLC to shift towards a fibroblast phenotype.     

Detection of bladder cancer by multitarget multicolour FISH: comparative analysis on archival cytology and paraffin-embedded tissue

Detection of bladder cancer by multitarget multicolour FISH: comparative analysis on archival cytology and paraffin-embedded tissue We have evaluated the possibility of using the same specimen for both cytological diagnosis and multitarget multicolour FISH (MtMcFISH) analysis in order to determine whether the routinely processed specimens used for diagnosis were also suitable for this ancillary procedure. For this purpose 18 positive samples (11 voided urine and seven bladder washings) were selected, together with a representative section of the corresponding immediately previous or subsequent histological specimens. Two negative cytology slides were added as negative controls. FISH analysis revealed a normal pattern for each probe in the two negative controls and an abnormal pattern in the 18 positive cases. In the latter the same FISH alterations were found in the cytology samples and in the corresponding histological sections, and superimposable cytological/histological features were observed in two cases where two different histology samples were analyzed. The results clearly show that MtMcFISH may be successfully applied to destained routinely processed cytology slides.     

Distribution and Relative Size of the Swim Bladder in Percina, With Comparisons to Etheostoma, Crystallaria, and Ammocrypta (Teleostei: Percidae)

The distribution of the swim bladder was examined in 20 species representing all four genera (Percina, Etheostoma, Crystallaria, and Ammocrypta) of darters (Etheostomatini). Including data on species examined in other studies, swim bladders have been found in 15 of 17 species of Percina, but were absent in Crystallaria asprella, Ammocrypta pellucida, and the 18 species of Etheostoma examined. Relative swim bladder length in Percina was shown to be positively correlated with body size. Although not statistically significant, swim bladder size also correlates with habitat (riffle, run, or pool) and swimming behavior (midwater or benthic).