肛垫与性器官勃起组织同源的探讨

目的 提出了一个肛垫组织胚胎发生与性器官海绵体勃起组织同源的假说.肛垫与性器官海绵体皆起源于泄殖腔膜周围的原始间充质.当泄殖腔膜被分割为尿生殖膜与肛膜后,前者周围的间充质形成性器官海绵体的胚胎原基,后者周围的间充质形成肛垫的胚胎原基.在成体,肛垫具有勃起组织特征和血管神经配布与会阴同源等事实支持这一学说.     

The efficacy of sildenafil in different etiologies of erectile dysfunction

PURPOSE: The aim of this study was to evaluate the efficacy of sildenafil and success of treatment in particular etiological causes in erectile dysfunction lasting more than 3 months. MATERIAL AND METHODS: A total of 141 patients between 27 and 78 years old without any cardiac compromise, despite controversial, which precludes sildenafil (Viagra) treatment, were included in this study. All patients had only International Index of Erectile Capacity Form (IIEF) for pre-treatment evaluation and 50 mg sildenafil was started. Patients were assessed monthly for 6 months thereafter. Erectile capacity changes were questioned by IIEF on each follow-up and 100 mg sildenafil was given in patients without a response and monthly follow-up was scheduled. All patients had SMA-12, hormonal analyses and penile colour Doppler ultrasonography during the treatment course. The difference between IIEF score of each patient was displayed by Paired-t test and p-values less than 0.05 was applied as significant. RESULTS: The average beginning IIEF score of 141 patients was 11.80 +/- 0.47 [6-22], and increased to 20.70 +/- 0.62 [6-30] after a month of 50 mg sildenafil treatment. The mean increase was 75.4% and found to be significant (p = 0.000, p < 0.05). The average IIEF scores were recorded as 22.57 +/- 0.69 after 3, and 22.12 +/- 0.24 after 6 months. There was no difference between these values and 2nd month controls (P3 month = 0.5675, P6 month = 0.6138, p > 0.05). A positive response was recorded in 102 patients (72.3%) and 39 (27.7%) patients were unresponsive. Doubled doses of sildenafil (100 mg) was effective in additional 17 patients. After overall treatment, 119 (84.4%) patients had benefit from sildenafil. Penile Doppler ultrasonography displayed arterial insufficiency in 79 (56.03%), veno-occlusive dysfunction in 14 (9.93%), mixt vascular pathology in 14 (9.93%) patients. Normal ultrasonographic findings in 32 patients (22.7%) were classified as psychogenic dysfunction. Among the organic causes, sildenafil was found to be most effective in arterial insufficiency group. CONCLUSION: Sildenafil is a successful management modality in erectile dysfunction with minimal pre-treatment evaluation. As far as etiological causes concerned, sildenafil was found to be most effective in arterial insufficiency group and psychogenic group. The efficacy of sildenafil treatment has not been changed with the treatment time, since IIEF scores were stable during follow-up controls.     

不同年龄大鼠终板软骨细胞抗氧化能力研究

目的 :通过研究不同年龄大鼠的终板软骨细胞氧化应激的差异,探讨衰老对终板软骨细胞抗氧化能力的影响。方法:原代培养获取SD大鼠2个月、18个月来源的终板软骨细胞鉴定后,使用第三代细胞进行实验。实验分为两组,A组为2个月大鼠来源的终板软骨细胞,B组为18个月大鼠来源的终板软骨细胞。待A、B两组细胞贴壁后,通过β-半乳糖脱氢酶染色和RT-PCR对端粒酶长度进行检测,比较两组细胞衰老的差异;通过总抗氧化能力试剂盒检测两组细胞总抗氧化能力的不同。以浓度为200umol/L过氧化氢对细胞进行2.5h的处理,RT-PCR法检测处理前后两组细胞Ⅱ型胶原、蛋白聚糖、caspase-3及B淋巴细胞瘤-2(B-cell lymphoma-2,bcl-2)的m RNA表达情况,使用流式细胞仪检测两组细胞的凋亡情况。对Ⅱ型胶原、蛋白聚糖、caspase-3及bcl-2的相对表达量进行组间t检验,P0.05为差异有统计学意义。结果 :未使用过氧化氢处理时,A、B两组终板软骨细胞的在β-半乳糖脱氢酶染色、Ⅱ型胶原、蛋白聚糖表达及凋亡率无明显差异(P0.05),而B组在端粒酶的相对长度(T/S=10±2)及总抗氧化能力(0.72±0.18)较A组(T/S=135±5,1)显著降低(P0.05)。在相同条件的过氧化氢刺激后,B组终板软骨细胞的Ⅱ型胶原(0.60±0.16)、蛋白聚糖(0.75±0.22)表达明显低于A组细胞(P0.05),而凋亡率显著高于A组细胞(P0.05)。结论 :不同年龄大鼠终板软骨细胞衰老及抗氧化能力存在差异,衰老越明显的终板软骨细胞,总抗氧化能力越低,抵抗过氧化损伤的能力越差。     

Nucleic acids in articular cartilage from rabbits of different ages.

The joint cartilage of immature, adult and old rabbits was examined. The joint cartilage in adult and old rabbits contained less nucleic acids than that in young rabbits. The synthesis of DNA decreases with advancing age, while no difference was found in the concentration of nucleic acids between adult and old rabbits.     

兔脂肪干细胞的分离培养鉴定及成骨诱导分化研究

[目的]探讨在成骨诱导条件下兔脂肪干细胞的体外诱导分化情况：[方法]取3个月龄日本大耳白兔颈背部皮下脂肪，用Ⅰ型胶原酶消化获得细胞。Stro-1免疫细胞化学染色鉴定细胞性质；加入成骨诱导液后依次进行形态学、Ⅰ型胶原、碱性磷酸酶及钙盐沉积相关检测。[结果]（1）原代所获细胞Stro—1表达阳性？（2）在诱导条件下，Ⅰ型胶原、碱性磷酸酶及钙盐沉积均呈阳性表达。[结论]脂肪干细胞来源广、易获取、对机体创伤小，与骨髓间充质干细胞类似，经体外诱导后可实现成骨分化，为骨组织工程提供了一种新的种子细胞：     

Cellular mechanisms of the epilepsies: In vitro studies on human tissue

BACKGROUND AND PURPOSE: Animal models have provided very valuable data to specify the physiopathological mechanisms of the various forms of epilepsy. However, the question arises of knowing which of these experimental results are relevant to the human epileptic brain. The development of epileptic surgery makes it possible to directly study the functional properties of human brain tissue in vitro and to analyze the mechanisms underlying seizures and epileptogenesis. We review some of the results obtained over the last few years in our laboratory based on electrophysiological, immunocytochemical and molecular experiments conducted on human brain tissue. RESULTS: This review covers a number of the mechanisms of neuronal synchronizations generating epileptiform discharges, including the role of electrical synapses connecting the inhibitory interneurons, particularly in Taylor-type focal cortical dysplasia and the functional lability of GABAergic inhibition in epileptogenic human cortical tissue, which may sustain triggering and propagation of seizures. Some of these mechanisms have not been described in animal models. CONCLUSIONS: Studies on human tissue, when carefully designed, are necessary to validate the data collected on animal models and will continue to provide us with new and important information on the cerebral changes related to epilepsy. Moreover, these studies allow development of a class of antiepileptic drugs that have a completely new mechanism of action, which could be effective in the treatment of drug-resistant epilepsies.     

Repair of mandible defect with tissue engineering bone in rabbits

BACKGROUND: The aim of the present study was to investigate the effect of tissue engineering bone composed of bone marrow-derived osteoblasts and demineralized bone in repairing mandible defect. METHODS: Bone marrow-derived osteoblasts of 20 rabbits were cultured and seeded into scaffold of allogeneic demineralized bone to construct tissue engineering bone graft in vitro, which was used to repair the 10 x 5-mm bone defect made in the same rabbit mandible edge. Implant of demineralized bone alone was as the control. Rabbits were killed according to the schedule: five after 2 weeks, five after 4 weeks, five after 8 weeks, five after 12 weeks, and the implants were harvested for gross, radiographic, and histological observation. RESULTS: New bone formation at the margin region of defect and osteogenesis at the centre were observed in the implant of tissue engineering bone, and the bone formation pattern included osteogenesis, osteoconduction, and osteoinduction. In the implant of demineralized bone alone, the major bone formation pattern was 'creeping substitute'. CONCLUSIONS: The tissue engineering bone graft constructed by autogenous bone marrow-derived osteoblasts and allogeneic demineralized bone was better than demineralized bone alone in bone formation capability, which might be an ideal graft for bone defect repair.     

不同年龄段人前列腺外周带基质细胞表型特征的研究

目的:探讨体外培养的不同年龄段人前列腺外周带基质细胞的表型特征和生长特性差异,分析前列腺基质细胞微环境对老年男性前列腺癌促发的可能原因。方法:原代培养不同年龄段前列腺外周带基质细胞,利用免疫细胞化学技术分析细胞表型特征、透射电镜观察细胞超微结构、琥珀酸脱氢酶法观察细胞体外增殖以及流式细胞术检测细胞凋亡情况。结果:不同年龄段前列腺基质细胞均表达成纤维细胞标记物prolyl-4-hydroxy-lase,而平滑肌肌动蛋白(α-SMA)、肌间线蛋白(desmin)的表达随年龄逐渐增加,即α-SMA的阳性表达:年轻组vs老年组=(2.56±1.81)%vs(38.89±11.22)%,(P<0.01);而desmin阳性表达:年轻组vs老年组=(0.89±0.93)%vs(14.89±5.97)%,(P<0.01)。并且α-SMA和/或desmin阳性细胞形态相对宽大、扁平、多形性。超微结构观察发现,前列腺基质细胞内蛋白合成有关的细胞器随着年龄的增加不断增多,主要为粗面内质网和高尔基复合体。老年组前列腺基质细胞的增殖率低于年轻组(P<0.01);而两组基质细胞凋亡率均在1%～3%之间,组间差异无统计学意义。结论:前列腺外周带基质细胞中具有活性的肌成纤维细胞随年龄不断增加可能是老年男性前列腺癌高发和恶性进展的原因之一。     

Evidence for the involvement of endothelial nitric oxide synthase from smooth muscle cells in the erectile function of the human corpus cavernosum

Nitric oxide (NO) is an important mediator in the relaxation of cavernosal smooth muscle. The present study examines the existence and location of the constitutive isoform eNOS (endothelial NO synthase) accompanying the already substantiated neurogenic NOS (nNOS) in the human corpus cavernosum of men with and without erectile dysfunction. Activities of NOS enzymes were examined in specimens of 11 potent and nine long-term impotent patients by means of light and electron microscopy using NADPH-diaphorase staining and immunohistochemical eNOS-specific, smooth muscle actin-specific and nNOS-specific markers. Cavernosal smooth muscle shows a distinct expression of eNOS. In contrast to the weaker expression of eNOS and nitrinergic innervation found in larger veins, the small intracavernosal helicine arteries express large quantities of eNOS and possess a dense nitrinergic innervation. Long-term impotent patients display a broad heterogeneity in eNOS expression and nitrinergic innervation while no overall correlation between NOS expression and erectile function was observed. The expression of eNOS indicates eNOS as a main source of NO alongside nNOS. The differentiated localization of eNOS supports at least a role of this isoform in vascular regulation. Received: 21 April 1997 / Accepted 11 November 1997     

不同年龄人真皮多能干细胞的分离及体外分化特点

目的研究不同年龄人真皮多能干细胞的分离、鉴定及体外分化特点。方法切取胎儿、患儿、中青年患者、老年患者正常皮肤标本（1cm×1cm），采用干细胞培养基筛选原代细胞克隆并记录其数量和直径；检测各种标本来源细胞克隆的体外分化情况；行单个细胞克隆分化分析，并检测其相关蛋白的表达。结果胎儿、患儿、中青年患者、老年患者正常皮肤原代培养获得的细胞克隆数分别为（20．1±2．5）×10^2、（15．8±5．7）×10^2、（10．8±1．3）×10^2、（6．2±1．4）×10^2个，两两比较，差异有统计学意义（P＜0．01）；克隆直径分别为（83±12）、（55±10）、（46±12）、（42±8）μm，胎儿＞患儿＞中青年及老年患者（P＜0．05）。各种细胞克隆均能向神经元细胞、神经胶质细胞、平滑肌细胞和脂肪细胞分化，其中胎儿来源的细胞克隆有向神经元细胞分化的优势。培养1个月，胎儿、患儿、中青年患者、老年患者单个细胞克隆形成率依次为41．1％、25．5％、17．7％、15．2％；单个细胞克隆亦能同上多向分化，同时表达巢蛋白、纤维连接蛋白、原癌基因c-mye、信号转导和转录激活因子3、端粒体逆转录酶。结论采用干细胞培养基能够有效分离不同年龄人真皮中具有克隆增殖和多向分化潜能的干细胞，年龄对其的数量、增殖和分化有明显影响。     

Phenotype and chondrogenic differentiation of mesenchymal cells from adipose tissue of different species

Mesenchymal stem cells (MSCs) are multipotent cells capable of differentiating into several mesoderm lineages. They have been isolated from different tissues, such as bone marrow, adult peripheral blood, umbilical cord blood, and adipose tissue. The aim of this study was to analyze the differences in proliferation and phenotype of adipose tissue‐derived MSCs from three different species, and to evaluate their capacity to differentiate into chondrocytes in vitro. A comparative study of cultured human, rabbit, and sheep mesenchymal cells from adipose tissue was carried out, and the main morphological parameters, proliferative activity, and expression of surface markers were characterized. Proliferation and flow cytometry data showed species‐related differences between animal and human MSCs. Histological staining suggested that rabbit and sheep mesenchymal cells were able to differentiate into chondrocytic lineages. Human mesenchymal cells, though they could also differentiate, accomplished it with more difficulty than animal MSCs. These results could help to explain the differences in the chondrogenic capacity of sheep and rabbit MSCs when they are used as animal models compared to human mesenchymal cells in a clinical assay. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1499–1507, 2009     

NIR laser tissue welding of in vitro porcine cornea and sclera tissue

BACKGROUND AND OBJECTIVE: The objective of this study was to test the hypothesis that an near infrared (NIR) laser system (1,455 nm) in combination with a motorized translational stage to control the position and speed of the laser beam and a shutter to control the laser exposure to the tissue being welded could be used to successfully weld ocular tissues. STUDY DESIGN/MATERIALS AND METHODS: Seventy-five porcine corneas and 23 porcine scleral tissues were welded in vitro in this study. The welded tissues were examined using histopathology and tensile strength analysis. Eight different welding conditions were analyzed for porcine cornea and one for sclera tissues. The tensile strength of the welded groups was compared to a sutured cornea control group. RESULTS: The NIR laser welding system provides strong, full thickness welds and does not require the use of extrinsic dyes, chromophores, or solders. Mean weld strengths of 0.15-0.45 kg/cm(2) were obtained for the cornea and 1.01 kg/cm(2) for sclera welds. The native H(2)O in the ocular tissue serves as an absorber of the 1,455 nm radiation and helps to induce the welds. CONCLUSIONS: We conclude that an NIR laser system using an optimal laser radiation wavelength of 1,455 nm can effectively weld cornea and sclera tissue and that this laser tissue welding (LTW) methodology typically causes minimal disruption of tissue, and thus, avoids opacities and irregularities in the tissue which may result in decreased visual acuity. The optimization of a laser welding system that leads to a strong full thickness tissue bond without tissue destruction, an instant seal that promotes wound healing, and the absence of a continued presence of a foreign substance like a suture, is of considerable importance to the ophthalmology medical community. This need is especially apparent with respect to corneal transplantation and fixing the position of corneal flaps in Laser-Assisted In Situ Keratomileusis (LASIK), a laser procedure used to permanently change the shape of the cornea.     

不同浓度丁酸钠体外诱导树突状细胞分化的细胞表型研究

目的 探讨不同浓度丁酸钠诱导对体外培养树突状细胞(DC)细胞表型的变化规律.方法 通过粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和白细胞介素-4(IL-4)体外诱导的人外周血单个核细胞来源的DC,按不同丁酸钠浓度和不同诱导时间分组培养,流式细胞仪检测各组DC的细胞表型.结果 在丁酸钠诱导下,各组DC表面的成熟标志均显著下调,丁酸钠浓度越高下调越明显,随着培养时间的延长,各组DC表面的成熟标志均显著下调;但是细胞的凋亡率也同时上升.结论 丁酸钠在体外可以显著抑制DC的成熟过程,以丁酸钠浓度为0.75mmol/L和培养48h为最佳的诱导浓度和时间.     

Endothelial rehabilitation: the impact of chronic PDE5 inhibitors on erectile function and protein alterations in cavernous tissue of diabetic rats

BACKGROUND: Diabetic men generally have reduced efficacy with PDE-5 inhibitors (PDE5i) for the treatment of erectile dysfunction (ED). OBJECTIVE: To determine whether chronic vardenafil exposure alters cavernous protein expression predicting improved erectile function in diabetes. DESIGN: Forty-two adult male Sprague Dawley rats with streptozotocin-induced (50mg/kg IP) diabetes for 4 wk, were exposed to either vehicle or vardenafil for 6 wk. Assessments compared the impact of vardenafil given at 1h and 20 h to erectile function and cellular alterations and downstream translation of cavernous protein profiles were aimed. INTERVENTION: Vehicle or vardenafil 0.5mg/kg/day by oral gavage for 6 wk. MEASUREMENTS: Erectile function, penile tissue morphology, protein expression and surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI) protein profiling were determined. RESULTS AND LIMITATIONS: A significant increase of intracavernous pressure was seen in both treatment arms compared to diabetic rats not receiving vardenafil. Immunohistochemical staining showed improved endothelial and smooth muscle cell staining with chronic vardenafil use. Western blot analysis demonstrated increased endothelial cell eNOS and smooth muscle alpha-actin protein content. SELDI protein profiling showed enhanced proteins expression at molecular weights of 14.7, 20, 41.9, 66.2, and 83.9 kDa in the chronically treated vardenafil group. CONCLUSIONS: Vardenafil was effective in treating diabetic-induced ED with the greatest effect achieved through chronic dosing, with no additive effect measured with the final acute dose. Changes noted in the histology and protein expression indicate that vardenafil may have a protective effect in this disease state. This finding may serve as a basis for further work evaluating the utility of chronic vardenafil dosing in diabetic men.     

Mechanical properties of brain tissue related to oedema development in rabbits

Summary We studied the mechanical properties of the brain in anaesthetized rabbits by application of a standard external load to the exposed cerebral surface. The experimental model used allows one to eliminate potential circulatory factors. Brain oedema was produced by repeated episodes of ischaemia secondary to a decrease of the arterial blood pressure to zero. The development of brain oedema was assessed by an increase of the cerebral water content. In the course brain oedema development brain fluidity was found to steadily rise, while the brain compliance and the index of hysteresis decreased from the control value found at the onset of the experiment. Most important both, brain compliance and the index of hysteresis were already markedly elevated prior to the manifestation of brain oedema.     

Direct measurement of acetylcholine release in detrusor smooth muscles isolated from rabbits

In the present study, we measured acetylcholine (ACh) released from rabbit detrusor smooth muscle strips induced by electrical field stimulation (EFS) using high-performance liquid chromatography coupled with microdialysis procedure. There were frequency- and duration-dependent increases in contractile response and ACh release. There was a significant, but not simple correlation between EFS-induced contraction and ACh release. Atropine caused a decrease and increase in the contractile response and ACh release, respectively. Pretreatment with propranolol increased ACh release, but pretreatment with phentolamine had no significant effect. These results demonstrate that this method is applicable to direct measurement of ACh release by EFS, and that neurotransmitters other than ACh may relate to EFS-induced contraction. In addition, it is suggested that there are prejunctional inhibitory muscarinic receptors and beta-adrenoceptors, which contribute to ACh release induced by EFS in the rabbit detrusor smooth muscles. Received: 20 January 1998 / Accepted: 17 April 1998     

Systemic and cavernous plasma levels of endothelin (1-21) during different penile conditions in healthy males and patients with erectile dysfunction

 The role of the sympathetic adrenergic nerves in mediating the constant tone of penile flaccidity and returning the erect penis to its flaccid state is fairly well established. However, it is not yet known whether additional nonadrenergic transmitters are involved in this process. The peptide endothelin-1 (ET-1) may be one of the factors contributing to such a control. Moreover, it has been speculated by various authors that ET-1 might be involved in the pathophysiology of erectile dysfunction. The present study was undertaken to determine whether or not there is a difference in the courses of ET-1/-2 plasma levels registered in systemic and cavernous blood cavities taken from healthy males and patients with ED during different penile conditions (flaccidity, tumescence/rigidity, detumescence). Thirty-two healthy adult males and 25 patients were exposed to visual and tactile erotic stimuli in order to elicit penile tumescence and, in the group of healthy volunteers, rigidity. Whole blood was aspirated from the corpus cavernosum and the cubital vein, and ET-1/-2 was determined in plasma aliquots by means of an enzyme-linked immunoassay (ELISA). Mean systemic and cavernous plasma level of ET-1/-2 in blood samples obtained from the volunteers was 0.2–0.7 fmol/ml. In the healthy males, no changes in ET-1/-2 levels were observed in the systemic and cavernous blood during penile tumescence, rigidity and detumescence. In the group of patients, mean plasma ET-1/-2 levels in the phase of penile flaccidity and detumescence were found to be higher in the systemic circulation than in the cavernous blood (flaccidity: 0.52 ± 0.38 fmol/ml vs. 0.48 ± 0.46 fmol/ml, respectively; detumescence: 0.53 ± 0.33 fmol/ml vs. 0.27 ± 0.11 fmol/ml, respectively). No differences in the plasma courses of ET-1/-2 were found between patients with an organogenic and psychogenic etiology of ED. In the phase of detumescence, the mean ET-1/-2 level was lower in the cavernous blood cavities taken from the patients than in the samples obtained from the healthy males. Our study revealed a difference in the profiles of ET-1/-2 registered in the cavernous blood of healthy subjects and patients with erectile dysfunction. Nevertheless, since this difference seems to be of no physiological significance, our data counteract the hypothesis of an ultimate importance of ET-1 in the control of penile flaccidity and detumescence and do not support speculations regarding an involvement of ET-1 in the pathophysiology of erectile dysfunction.     

不同年龄神经再生组织特异性差异的实验研究

目的　比较不同年龄大鼠坐骨神经切断后神经再生趋化性在组织特异性水平上的差异。方法　SD大鼠40只,按年龄分成幼年组和成年组,每组20只,建立组织特异性模型。右侧坐骨神经切断后,将神经近端接于Y形硅胶管入口端,神经远端和肌腱分别接于Y形管两出口端。术后3、6周采用神经电图和组织学方法检测神经再生情况。结果　术后3、6周,幼年组实验侧运动神经传导速度的恢复率分别为(27.21±6.06)%和(39.33±10.41)%,成年组分别为(38.23±11.46)%和(52.36±8.12)%;两组的差异有显著性和非常显著性意义（F=6.49、10.73,P＜0.05、P<0.01）。幼年组复合肌肉动作电位（CMAP）波幅的恢复率分别为(30.21±17.43)%和(30.42±18.43)%,成年组分别为(35.43±32.13)%和(37.18±16.12)%;两组的差异无显著性意义（F=0.23、0.92,P>0.05）。幼年组神经纤维再生准确率分别为(59.31±13.21)%和(59.39±24.16)%,成年组分别为(78.26±12.41)%和(84.28±9.01)%;两组的差异有非常显著性意义（F=10.75、8.36,P<0.01）。结论　周围神经损伤后,远端神经组织对再生轴索有诱导作用,但幼年大鼠神经再生趋化的组织特异性较成年大鼠差。