Morphometric and stereological study of the glandular epithelium of the lateral prostate of the intact and castrated guinea pig

The glandular epithelium of the lateral prostate of the guinea pig was described within the framework of a morphometric model in terms of relative densities and absolute dimensions. A combination of direct measurement and point and intersection counting techniques was used. The quantitative data generated in the intact animals were compared with those of castrated controls. Castration was accompanied by a significant decrease in height of the glandular epithelium and in sizes of secretory and basal cells and their corresponding nuclei. On a per cell basis, significant decreases in total volume and surface area of granular endoplasmic reticulum were detected after castration. This was accompanied by a significant reduction in the total volume of Golgi cisternae. The total volume, surface area, and number of highly electron-dense and clear granules decreased significantly compared with the intact control animals. However, no significant changes in these parameters of low electron-dense granules were found. Significant reductions in the total volume and surface area of condensing granules, lysosomes, and mitochondria, but not their number, were detected. The average sizes of condensing granules, secretory granules, lysosomes, and mitochondria were decreased significantly after castration. The present study showed that the alterations in the secretory function of the secretory cells of the lateral prostate was reflected by the quantitative changes in granular endoplasmic reticulum, Golgi complexes, and secretory granules on a per cell basis. The data generated in the present study will serve as a baseline for further studies of the lateral prostate of the guinea pig.     

Stem cells: novel players in the treatment of erectile dysfunction

Stem cells are defined by their capacity for both self-renewal and directed differentiation; thus, they represent great promise for regenerative medicine. Historically, stem cells have been categorized as either embryonic stem cells (ESCs) or adult stem cells (ASCs). It was previously believed that only ESCs hold the ability to differentiate into any cell type, whereas ASCs have the capacity to give rise only to cells of a given germ layer. More recently, however, numerous studies demonstrated the ability of ASCs to differentiate into cell types beyond their tissue origin. The aim of this review was to summarize contemporary evidence regarding stem cell availability, differentiation, and more specifically, the potential of these cells in the diagnosis and treatment of erectile dysfunction (ED) in both animal models and human research. We performed a search on PubMed for articles related to definition, localisation and circulation of stem cells as well as the application of stem cells in both diagnosis and treatment of ED. Strong evidence supports the concept that stem cell therapy is potentially the next therapeutic approach for ED. To date, a large spectrum of stem cells, including bone marrow mesenchymal stem cells, adipose tissue-derived stem cells and muscle-derived stem cells, have been investigated for neural, vascular, endothelial or smooth muscle regeneration in animal models for ED. In addition, several subtypes of ASCs are localized in the penis, and circulating endogenous stem cells can be employed to predict the outcome of ED and ED-related cardiovascular diseases.     

Morphometric analysis of the components of the neonatal and the adult rat testis interstitium

Leydig cells in the foetal rat testis are still present at birth and it has been hypothesized that they commence to degenerate immediately after birth, based on the decrease in their volume density (v/v%) with age. In this study the interstitium of the rat testis was studied quantitatively at 1, 5, 10, 15, 20 and 90 days after birth: the latter are considered to be adults. The absolute volumes of connective tissue cells and blood vessels increased with age. The absolute volumes of macrophages and lymphatic spaces were greater at 90 days than at any other age. The absolute volume of foetal Leydig cells per testis was unchanged from 1 to 15 days, despite a decrease in the % volume occupied per testis. The number of foetal Leydig cells per testis did not decline from days 1-20 although on day 20 an average foetal Leydig cell was smaller in volume than at earlier ages (days 1-15). Adult Leydig cells were recognized at day 10 and their absolute volume and number per testis increased from 15 to 90 days. Adult Leydig cells were similar in morphology to foetal Leydig cells at 20 days except for a reduced volume of cytoplasmic lipid.     

Correlations between the Quantitative Morphology of the Human Testis and Sperm Production The Testis of Healthy Men with Normal Sperm Counts

Stereology was used as a morphometric method to study the testicular biopsies of 8 healthy volunteers with a proven fertility and normal sperm counts.
The geometrical mean of the sperm density and total sperm count of 2 semen samples were correlated with the measured and calculated stereological parameters. The volume density of the germinal cell nuclei ( r = 0.72) and the intra-tubular tissue volume density ( r = 0.81) were significantly correlated with the sperm density. A significant correlation was demonstrated between the total sperm count and the volume density of the germinal cell nuclei ( r = 0.71) and the volume density of the Leydig cells ( r =–0.70).
Using stepwise multiple regression analysis the prediction of the observed variance of the log sperm density was feasible ( r = 0.996; P = 0.0007) with 4 stereological parameters: the volume density of the germinal cell nuclei (positive correlation), the volume density of the tubular wall (negative correlation), the volume density of the Leydig cells (negative correlation) and the tubular surface density (positive correlation).
The observed variance of the log total sperm count was predicted ( r = 0.96; P = 0.003) with 3 stereological parameters: the volume density of the germinal cell nuclei (positive correlation), the volume density of the Leydig cells (negative correlation) and the volume density of the blood vessels (negative correlation).     

肿瘤干细胞对恶性肿瘤诊治的意义

恶性肿瘤严重地威胁着人类的健康.多药耐药现象增加了其治疗难度,局部复发和远处转移最终导致了治疗失败,然而遗憾的是机制尚不清楚.研究发现在肿瘤组织中有一个具有自我更新及分化潜能的的细胞亚群,是复发及转移的根源.这部分细胞被称为肿瘤干细胞.随后,人们相继从血液系统肿瘤及多种实体瘤中分离、鉴定了肿瘤干细胞.随着肿瘤干细胞学说的提出及确认,人们对肿瘤的复发、转移及多药耐药现象有了新的认识,从而为恶性肿瘤的治疗及复发转移的预防提供了一个全新的线索.本文就此进行综述.     

血管紧张素Ⅱ对骨髓间充质干细胞定向诱导为角质形成细胞调控作用的实验研究

目的：探讨骨髓间充质干细胞（MSCs）分化为角质形成细胞的可能性及在此过程中血管紧张素Ⅱ （AngⅡ）对其的调控作用.方法：抽取Wistar大鼠的骨髓,经全骨髓法分离、纯化MSCs,鉴定后建立MSCs细胞模型,用成胶质细胞诱导培养基诱导其为角质形成细胞,显微镜下观察其形态学变化.以添加了AngⅡ 的成角质形成细胞诱导培养基组与单纯成角质形成细胞诱导培养基组在诱导MSCs 7天、10天后行角蛋白10（CKP10）免疫组织化学染色及流式细胞仪检测,并进行对比观察分析.结果：培养的MSCs具有成脂、成骨分化的能力.MSCs可以成功诱导为角质形成细胞,添加Ang Ⅱ的诱导组与对照诱导组CKP10免疫组化染色均有阳性表达,但添加AngⅡ组阳性细胞数高于对照组.流式细胞仪检测显示CKP10阳性细胞百分率,AngⅡ组为80.62％,对照组（32.46％）,两者相差显著（P＜0.05）.结论：MSCs可以诱导分化为角质形成细胞,ANGⅡ对MSC的成角质形成细胞分化有显著的促进作用,这一作用可能是AngⅡ 促进创面愈合的机制之一.     

不同组织来源间充质干细胞体外成骨分化能力的比较研究

目的比较成人脂肪间充质干细胞(ASCs)、脐带间充质干细胞(UC-MSCs)和成人骨髓间充质干细胞(BMSCs)的成骨能力,选择优势干细胞种类作为应用于骨组织工程治疗骨缺损的种子细胞。方法采用含10%胎牛血清的DMEM/Ham’s F-12培养液培养3种MSCs。取3种MSCs的P3代,通过CCK8方法检测其增殖能力;通过流式细胞仪进行鉴定;通过碱性磷酸酶(ALP)和茜素红染色检测骨分化蛋白ALP的分泌和矿化钙结节的沉积,并对钙结节进行定量分析;通过实时荧光定量PCR(RT-q PCR)方法检测骨再生相关基因的表达。结果 3种P3代MSCs在3~5d之间增殖均处于对数生长期;流式鉴定3种细胞的表面标志物阳性率:CD44、CD90和CD105均高于97%,阴性率:CD14、CD34和CD45均低于1%;ALP染色结果显示3种MSCs成骨诱导9d时,细胞内均表达ALP,茜素红染色结果显示成骨诱导18d时,均呈现较好的矿化能力,BMSCs和UC-MSCs成骨诱导后形成的钙结节无显著性差异;RT-q PCR结果显示3种MSCs成骨诱导组相比较于对照组,成骨再生相关基因Osterix、ALP、I型胶原(COL1)和骨钙素(OCN)均显著性高表达;3种MSCs成骨诱导9d时,UC-MSCs实验组的COLI基因表达显著性高于BMSCs,成骨诱导18d时,ASCs实验组的Osterix基因表达显著性高于BMSCs。结论 ASCs和UC-MSCs具有一定的成骨矿化能力,有望成为骨组织工程治疗骨缺损的种子细胞。     

低氧对脂肪间充质干细胞向雪旺细胞分化的影响

目的探讨低氧对脂肪间充质干细胞（ADMSCs）向雪旺细胞（SCs）分化能力的影响。方法分离培养SD大鼠ADMSCs并用流式细胞仪、茜素红染色、油红O染色鉴定。将成功分离的ADMSCs随机分为3组：常氧诱导组,在常氧条件下（5％CO2,21％O2,37℃）诱导;低氧处理＋常氧诱导组,低氧处理（5％CO2,0．5％O2,37℃）后在常氧条件下诱导;低氧诱导组,低氧条件下（5％CO2,0．5％O2,37℃）诱导。观察各组细胞形态,MTT法检测细胞增殖情况,免疫荧光染色和Westernblot检测SCs标志物GFAP和S-100的表达。结果细胞分离后,经流式细胞仪分析可见细胞表面CD44阳性、CD45阳性、CD90阳性,茜素红及油红O染色均为阳性。MTT法检测结果：低氧处理＋常氧诱导组A值为0．861±0．039,高于常氧诱导组0．837±0．017,差异具有统计学意义（P〈0．05）;低氧诱导组A值为0．931±0．041,均高于常氧诱导组和低氧处理＋常氧诱导组（P均〈0．05）。免疫荧光染色发现常氧诱导组和低氧处理＋常氧诱导组大量细胞GFAP和S-100表达阳性,低氧诱导组仅少量细胞S-100和GFAP表达阳性。Westernblot检测发现常氧诱导组S-100蛋白表达最高,低氧处理＋常氧诱导组GFAP蛋白表达最高,低氧诱导组S-100蛋白、GFAP蛋白表达均最低。结论低氧抑制ADMSCs向SCs的分化,低氧处理后的ADMSCs在常氧条件下仍可向SCs分化。     

大鼠嗅鞘细胞无血清上清液诱导脊髓神经干细胞分化的研究

[目的]观察大鼠嗅神经鞘细胞上清液对脊髓神经干细胞共培养发生的诱导分化作用。[方法]采用差速贴壁法获得较高纯度的嗅鞘细胞，分时段Mr丌法检测细胞活性，选取最佳状态细胞无血清培养后取上清液，与3代纯化后的神经干细胞共培养，观察分化过程，免疫荧光法鉴定诱导结果。[结果]MTT法分6个时段检测纯化后的嗅鞘细胞，发现9d及12d细胞活性最高。使用无血清嗅鞘细胞上清液与脊髓神经干细胞共培养发现诱导作用明显，向神经元样细胞及胶质细胞分化的比例分别达到53％和42％。[结论]嗅鞘细胞在体外培养的不同时间段活性不同，无血清的嗅鞘细胞上清有明显诱导神经干细胞向成熟神经元分化的作用。     

Activation of T lymphocytes for the adoptive immunotherapy of cancer

Background: Adoptive immunotherapy of malignancy involves the passive transfer of antitumor-reactive cells into a host in order to mediate tumor regression. Based on animal models, the transfer of immune lymphoid cells can eradicate widely disseminated tumors and establish long-term systemic immunity. Critical for successful adoptive immunotherapy is the ability to isolate large numbers of immune cells. For clinical therapy, it will require the development of in vitro methods to promote the sensitization and propagation of tumor-reactive cells. However, this is formidable task since human cancers are postulated to be poorly immunogenic because of their spontaneous origins. Results: Human lymphoid cells for ex vivo activation and subsequent adoptive transfer have been derived from different sources, including peripheral blood, tumor, and lymph nodes. Peripheral blood lymphocytes can be incubated with interleukin 2 to generate lymphokine-activated killer (LAK) cells, which nonspecifically lyse autologous and allogeneic tumor cells in vitro. LAK cell therapy represented the earliest attempt to treat advanced human cancers, with encouraging results documented in patients with renal cell cancer and melanoma. From the experience, the use of more immunologically specific cellular agents with potentially greater therapeutic efficacy has been investigated. One approach uses tumor-infiltrating lymphocytes, which have been characterized experimentally to be more specific in tumor reactivity compared with LAK cells. Other techniques have involved the use of lymphoid cells derived from lymph nodes draining tumors or primed by tumor vaccines. In vitro activation of these cells with tumor antigen or anti-CD3 monoclonal antibody results in the generation of T cells that mediate the rejection of poorly immunogenic tumors in animal studies. These alternate methods are currently being evaluated in clinical studies. Conclusions: Experimentally, cellular therapy is a potent method to eradicate progressive tumors. Initial clinical studies have demonstrated that this form of therapy is technically feasible and can result in meaningful antitumor responses. Advances in this area will require improved methods to sensitize, isolate, and expand tumor-reactive T cells for adoptive transfer. Presented at the 46th Annual Cancer Symposium of The Society of Surgical Oncology, Los Angeles, California, March 18–21, 1993.     

肠上皮干细胞分离培养方法研究进展

肠上皮干细胞是位于肠黏膜陷窝内的具有自我更新和增殖分化为成熟肠上皮细胞功能的细胞。近年来,肠上皮干细胞的研究越来越受到重视,通过分离SP细胞、培养隐窝器官样组织和利用组织工程等方法研究肠上皮干细胞的分裂、分化过程,加深了对肠上皮干细胞的生物学特征的了解。但肠上皮干细胞的分离、培养和鉴别等研究方法尚未取得突破性进展。     

Evaluation of the effect of peritubular cell secretions and the testicular paracrine factor P-Mod-S on Leydig cell steroidogenesis and immunoactive inhibin production

Testicular peritubular cells have been shown to produce a paracrine factor, termed P-Mod-S, under androgen control that has dramatic effects on Sertoli cell function and may provide an important mode of androgen action in the testis. Therefore, the current study was designed to investigate the possibility that peritubular cell secretory products could feedback and regulate Leydig cell function. The Leydig cell functional parameters that were examined included testosterone production and inhibin secretion. Purified forms of P-Mod-S (P-Mod-S(A) and P-Mod-S(B) shown to be biologically active on Sertoli cells) had no effect on basal or gonadotrophin-stimulated production of testosterone or inhibin by Leydig cells. A preparation of peritubular cell-secreted proteins (PSP) with molecular weights greater than 3 kDa did not influence testosterone production by Leydig cells. PSP, however, did influence cultured Leydig cell morphology and improved cell viability. PSP also had no effect on the ability of LH to stimulate Leydig cell testosterone production. Whilst determining the effect of PSP on Leydig cell inhibin production, PSP was found to contain endogenous levels of inhibin apparently due to 2% contamination of the peritubular cell cultures with Sertoli cells. When this endogenous inhibin level was considered, PSP was found to have no influence on basal or hormone-stimulated production of inhibin by Leydig cells. Results of the current study indicate that peritubular cell secretory products, including the paracrine factor P-Mod-S, do not appear to play a major role in the regulation of Leydig cell function. Therefore, the regulation of Leydig cell function by the seminiferous tubule will primarily be due to Sertoli cell secretory products.     

Correlative Study of the Ultrastructure and the Physiology of the Seasonal Regression of the Epididymal Epithelium in the Hedgehog Paraechinus Micropus

Korrelierende Studie der Ultrastruktur und der Physiologie der jahreszeitlichen Rückbildung des Nebenhodenepithels beim Igel Paraechinus micropus Die jahreszeitlichen Schwankungen der Ultrastruktur und Physiologie des Nebenhodens beim Igel wurden in bezug auf die Reproduktionsfunktion untersucht. 5 ausgewachsene männliche Igel wurden während eines Jahres einen über den anderen Monat lang getötet. Der Nebenhoden wurde für die Lichtmikroskopie fixiert in Bouin's Zenker und Formalin-Calcium, in für die Elektronenmikroskopie kaltgepuffertem Glutaraldehyd, dann in Osmiumtetraoxid. Das Nebenhodenepithel setzt sich aus vier Zellarten zusammen: der Hauptzelle, der Apicalzelle, der Dunkel- und Basalzelle. Die Hauptzellen enthalten - wie andere Steroide synthetisierende Zellen - einen ausgedehnten Golgiapparat, das glatte und rauhe endoplasmatische Retikulum, die sekretorischen Bläschen und die Lipidgranula während der Brutzeit, aber praktisch fehlen alle diese Zellorganellen während der Regression, abgesehen von einem verzögerten Golgiapparat und einem schwach ausgeprägten rauhen endoplasmatischen Retikulum. Andererseits zeigen sich in den Basalzellen während der Regression Lipide und gut entwickelte Organellen, jedoch schlecht entwickelte Strukturen beim sexuell aktiven Igel. Möglicherweise üben sie eine Funktion aus, da die Zellen während der Regression Lipide speichern, die später bei der Reaktivierung nach dem Winter-schlaf benötigt werden. Das Nebenhodenepithel erneuert sich gemeinsam mit dem Epithel der Samenkanälchen bevor die Spermatozoen das Nebenhodenlumen erreichen, wohingegen die akzessorischen Geschlechtsdrüsen, die auch zu den androgenabhängigen Organen außerhalb der Hoden gehören, noch zurückgebildete Strukturen zeigen. Somit geht aus den ultrastrukrurellen und physiologischen Beobachtungen hervor, daß die Hauptzellen möglicherweise der Ort der Androgensynthese sind und sie vollständig entwickelt werden gemeinsam mit den Zellen des Hodens nach Stimulation aus der Hirnanhangdrüse zu Beginn des Wieder-erwachens.     

新鲜分离的脂肪SVF细胞促进脂肪移植存活的实验研究

目的 探讨应用自脂肪组织新鲜分离的血管基质层细胞辅助脂肪移植,提高移植物存活率的可行性.方法 将0.5 ml待移植的脂肪颗粒分别与下列细胞混合:①DiI标记的新鲜分离的自体血管基质层细胞(A组);②DiI标记的培养至第4代自体脂肪来源干细胞(B组);③DMEM完全培养基(C组),随机注射移植于14只新西兰兔背部皮下.术后观察:①湿重;②切片HE染色计数血管密度;③方网测试系统"点计数"法检测存活脂肪细胞计数以及纤维组织计数;④荧光显微镜检测DiI标记的细胞在体内的分化转归.结果 ①湿重:A组(291.0±72.1)mg,B组(269.3±67.3)mg,C组(177.8±60.0)mg,A、B两组脂肪存活率均高于C组(P<0.05),两组之间比较差异无统计学意义(P>0.05);②血管密度:A、B两组血管密度均高于C组(P<0.05),两组之间比较差异无统计学意义(P>0.05).③点计数:A、B两组存活脂肪细胞计数均高于C组(P<0.05),纤维组织计数均低于C组(P<0.05),两组之间比较差异均无统计学意义(P>0.05);④荧光显微镜下观察发现自体血管基质层细胞与自体脂肪来源干细胞在体内均可向血管内皮细胞分化.结论 自体血管基质层细胞与培养的自体脂肪来源于细胞均可提高脂肪移植物存活率,但前者操作更方便,安全性更高,具有广阔的临床应用前景.     

周细胞参与婴幼儿血管瘤消退机制的研究进展

婴幼儿血管瘤是儿童期最常见的肿瘤,具有特殊的自然周期。周细胞与内皮细胞共同参与其发生、发展与消退的演变过程。本文通过总结周细胞的来源、分化,及其与内皮细胞的关系,对周细胞在婴幼儿血管瘤消退过程中的作用进行综述。     

Cell proliferation studies in the rat prostate: II. The effects of castration and androgen-induced regeneration upon basal and secretory cell proliferation

The changes over short and prolonged periods (up to three months) after castration on the proliferative activity of basal and secretory epithelial cells in the rat prostate were studied. Although castration induced widespread apoptosis of the secretory cells, no compensatory hyperplasia of the basal cells in response to this was noted. Instead, observations of the cell kinetics and ultrastructure suggested that both the basal and secretory cells entered a quiescent state as a result of castration. The proliferative potential of secretory cells was not diminished up to three months after castration. During androgen-induced regeneration of the prostate the pattern of basal and secretory cell proliferation was found to be similar to that observed during normal growth, although it was more rapid and of shorter duration.     

分层共同培养诱导骨髓间充质干细胞向表皮细胞分化的研究

目的 探讨体外联合HaCaT细胞共同培养诱导骨髓间充质干细胞(MSCs)向表皮细胞分化的可行性.方法 用聚碳酸酯细胞插入板分层后联合共同培养HaCaT细胞与MSCs,观察培养3、6、9d后的细胞形态,进行角蛋白(CK-19、CK-10)、整合素(α6、β1)染色并用流式细胞仪统计细胞阳性率.结果 共同培养后细胞形态变化明显,共培养3、6d后表皮细胞标志物CK-19、α6整合素、β1整合素免疫荧光染色阳性,细胞阳性率分别为9.3％、8.2％、11.5％和21.7％、34.1％、39.6％,CK10表达呈阴性;共培养9d后CK-19、α6整合素、β1整合素、表达较前减少,阳性率为12.2％、18.6％、16.3％,CK1O出现阳性表达,细胞阳性率为10.7％.结论 分层联合HaCaT细胞共同培养可以诱导骨髓干细胞向表皮细胞进行分化.     

Multinucleate epithelial cells in the ductuli efferentes of human epididymis: Mehrkernige Epithelzellen in den Ductuli efferentes des menschlichen Nebenhodens

The histological and ultrastructural study of the ductuli efferentes in epididymides from 40 adult men revealed the occurrence of multinucleate epithelial cells in all specimens. These cells appeared in the luminal protrusions of epithelial folds and correspond to either principal or ciliated cells. The ultrastructure of their cytoplasm did not differ from that of their respective mononucleate cells. Multinucleate cells contained 3-20 closely juxtaposed nuclei, thus appearing much more irregularly outlined than those of the mononucleate epithelial cells. Multinucleation four times more frequent in the principal cells than in the ciliated cells. The number of multinucleate cells increased progressively from the age of 60 years onwards. The average number of nuclei per cell increased in the fourth decade of life, was maintained up to the eighth decade, and then increased again.