低氧对培养的不同内径的肺动脉平滑肌细胞增殖的影响

目的和方法：分离培养三种不同内径的肺动脉平滑肌细胞（PASMCs）,用^3H－TdR掺入速率和细胞计数作为细胞增殖的指标,观察低氧对其增殖作用的影响。结果：低氧对三种不同内径的PASMCs（内径分别为＞1000μm、500－800μm、300-400μm）增殖促进作用显著不同,其^3H－TdR掺入速率和细胞计数分别增加23.5％和11.1％、60.0％和33.8％、141.4％和52.0％,选择对低氧最敏感的PASMCs（内径为300－400μm）,进一步探讨低氧促PASMCs增殖作用的细胞机制：钙拮抗剂verapail、蛋白激酶C抑制剂staurosporine(Stau)和细胞Na-H交换抑制剂amiloride可显著降低低氧情况下PASMCs^3H－TdR掺入速率和细胞计数。结论：低氧对三种不同内径的PASMCs增殖促进作用显著不同; Ca^2 、蛋白激酶C和Na^2 －H^ 交换的激活,可能是低氧促PASMCs增殖的重要胞内信息转导机制。     

Estrogen-alpha and progesterone receptor expression in cystic endometrial hyperplasia and pyometra in the bitch

Estrogen-alpha receptor (ER) and progesterone receptor (PR) were examined immunohistochemically in uteri of normal bitches, in uteri of bitches with cystic endometrial hyperplasia-mucometra (CEH-M) and in uteri of bitches with endometritis-pyometra (E-P), under exogenous progesterone treatment.In the CEH-M group, the ER- and PR-scores of all uterine cell types were higher than the ER- and PR-scores of normal uteri, although these differences were not always statistically significant. The ER-scores of E-P group were significantly lower than the ER-scores of the normal uteri and CEH-M group. The PR-scores of the E-P group tended to be higher than the PR-scores of the normal uteri, except for the surface epithelium, although these differences were not statistically significant. Exogenous progesterone treated bitches with CEH-M or E-P showed reduced ER- and PR-scores in the different uterine cell types, compared with the corresponding nontreated CEH-M or E-P group.The differences in ER and PR expression between CEH-M and E-P suggest different factors in the pathogenesis of both entities. Although, these changes in ER and PR expression do not seem to be directly involved in the pathogenesis of CEH-M and E-P. It is suggested that for CEH-M and progestin induced CEH-M a hormone dependent pathway is responsible. For P, the trigger may be bacterial infection.     

Role of stroma in carcinogenesis of the prostate

Prostatic development is induced by androgens acting via mesenchymal-epithelial interactions. Androgens elicit their morphogenetic effects by acting through androgen receptors (ARs) in urogenital sinus mesenchyme (UGM), which induces prostatic epithelial development. In adulthood reciprocal homeostatic stromal-epithelial interactions maintain functional differentiation and growth-quiescence. Testosterone plus estradiol (T+E2) have been shown to induce prostatic carcinogenesis in animal models. Thus, tissue recombinant studies were undertaken to explore the mechanisms of prostatic carcinogenesis in BPH-1 cells in which ARs and estrogen receptors (ERs) are undetectable. For this purpose, BPH-1 cells were combined with UGM, and the UGM+BPH-1 recombinants were grafted to adult male hosts. Solid branched epithelial cords and ductal structures formed in untreated UGM+BPH-1 recombinants. Growth was modest, and tumors did not develop. UGM+BPH-1 recombinants treated with T+E2 formed invasive carcinomas. BPH-1 cells lack ARs and ERs, whereas rat UGM expresses both of these receptors. These data show that immortalized nontumorigenic human prostatic epithelial cells can undergo hormonal carcinogenesis in response to T+E2 stimulation via paracrine mechanisms and demonstrate that the stromal environment plays an important role in mediating hormonal carcinogenesis. During prostatic carcinogenesis the stroma undergoes progressive loss of smooth muscle with the appearance of carcinoma-associated fibroblasts (CAF). This altered stroma was tested for its ability to promote carcinogenesis of nontumorigenic but immortalized human prostatic epithelial cells (BPH-1). CAF+BPH-1 tissue recombinants formed large carcinomas. In contrast, recombinants composed of normal prostatic stroma+BPH-1 cells exhibited minimal growth. This stroma-induced malignant transformation was associated with additional genetic alterations and changes in gene expression. Thus, alteration in the stromal microenvironment was sufficient to promote malignant transformation of human prostatic epithelial cells.     

Potassium ion channels and human disease: phenotypes to drug targets?

A significant difficulty faced by the pharmaceutical industry is the initial identification and selection of macromolecular targets upon which de novo drug discovery programs can be initiated. A drug target should have several characteristics: known biological function; robust assay systems for in vitro characterization and high-throughput screening; and be specifically modified by and accessible to small molecular weight compounds in vivo. Ion channels have many of these attributes and can be viewed as suitable targets for small molecule drugs. Potassium (K⁺) ion channels form a large and diverse gene family responsible for critical functions in numerous cell types, tissues and organs. Recent discoveries, facilitated by genomics technologies combined with advanced biophysical characterization methods, have identified novel K⁺ channels that are involved in important physiologic processes, or mutated in human inherited disease. These findings, coupled with a rapidly growing body of information regarding modulatory channel subunits and high resolution channel structures, are providing the critical information necessary for validation of K⁺ channels as drug targets.     

TURP联合经尿道膀胱颈切开术治疗小体积前列腺增生所致膀胱出口梗阻的疗效分析

目的:探讨经尿道前列腺电切术(TURP)联合经尿道膀胱颈切开术(TUIBN)治疗小体积前列腺增生(BPH)所致膀胱出口梗阻的疗效。方法:选择2009年1月~2013年12月我院收治的小体积BPH患者,其中单纯经尿道前列腺电切术(TURP组)48例,经尿道前列腺电切术联合经尿道膀胱颈切开术(TURP+TUIBN组)48例。比较两组的术前、术后国际前列腺症状评分(IPSS)、残余尿量(PVR)、最大尿流率(Qmax)等,以及术后并发症的发生情况。结果:TURP+TUIBN组术中出血量较TURP组明显增多(P0.05),两组手术时间、组织切除质量比较,差异均无统计学意义(P0.05);与TURP组比较,TURP+TUIBN组术后6个月IPSS评分、PVR明显下降,Qmax、膀胱压力明显上升(P0.05);TURP+TUIBN组并发症发生率为4.2%,显著低于TURP组16.7%(P0.05)。结论:TURP+TUIBN治疗小体积前BPH所致膀胱出口梗阻,可彻底切除增生腺体,消除小体积BPH的各种梗阻因素,减少术后膀胱颈挛缩的发生。     

Otoacoustic emissions,auditory evoked potentials and self-reported gender in people affected by disorders of sex development (DSD)

Both otoacoustic emissions (OAEs) and auditory evoked potentials (AEPs) are sexually dimorphic, and both are believed to be influenced by prenatal androgen exposure. OAEs and AEPs were collected from people affected by 1 of 3 categories of disorders of sex development (DSD) — (1) women with complete androgen insensitivity syndrome (CAIS); (2) women with congenital adrenal hyperplasia (CAH); and (3) individuals with 46,XY DSD including prenatal androgen exposure who developed a male gender despite initial rearing as females (men with DSD). Gender identity (GI) and role (GR) were measured both retrospectively and at the time of study participation, using standardized questionnaires. The main objective of this study was to determine if patterns of OAEs and AEPs correlate with gender in people affected by DSD and in controls. A second objective was to assess if OAE and AEP patterns differed according to degrees of prenatal androgen exposure across groups. Control males, men with DSD, and women with CAH produced fewer spontaneous OAEs (SOAEs) – the male-typical pattern – than control females and women with CAIS. Additionally, the number of SOAEs produced correlated with gender development across all groups tested. Although some sex differences in AEPs were observed between control males and females, AEP measures did not correlate with gender development, nor did they vary according to degrees of prenatal androgen exposure, among people with DSD. Thus, OAEs, but not AEPs, may prove useful as bioassays for assessing early brain exposure to androgens and predicting gender development in people with DSD.     

Validation of a total testosterone assay using high-turbulence liquid chromatography tandem mass spectrometry: Total and free testosterone reference ranges

Accurate measurement of testosterone concentration is of critical importance when diagnosing and treating male hypogonadism, congenital adrenal hyperplasia, premature or delayed puberty, and androgen excess in polycystic ovary syndrome or other virilizing conditions. However, some assays have inherent limitations and biases that affect measurement of low-testosterone values. Therefore, we developed a highly specific online mass spectrometry method. Sera were extracted online using high-turbulence flow liquid chromatography coupled to analytical HPLC and atmospheric pressure chemical ionization tandem mass spectrometry (HTLC-APCI-MS/MS). Analyte ions were monitored by multiple reaction monitoring (MRM). Total analysis time was 1.15 min per sample when using the multiplexing system. Testosterone concentrations were measured directly from 150 μL of serum or plasma without derivatization or liquid-liquid extraction. The lower limit of quantification was 0.3 ng/dL, and the assay was linear up to 2000 ng/dL. The method compared very well with an established RIA: y = 1.02x + 1.5, r² = 0.994. Comparison with a platform immunoassay confirmed the previously reported ICMA positive bias at low concentrations. Male and female adult and pediatric reference ranges were developed for this very sensitive and accurate high-throughput LC-MS/MS method. This method is suitable for measuring the expected low-testosterone concentrations seen in women, children, and hypogonadal males and for monitoring testosterone suppressive therapy in prostate cancer patients.     

Reaction-diffusion approach to modeling of the spread of early tumors along linear or tubular structures

We consider a system composed of a tubular sheet of early tumor cells, occupying the surface of a structure existing in the organism. We assume that the cells have a potential for proliferation in response to a growth factor. This model can be thought of as representing an early stage (pre-in situ) of tumor evolution. A biomedical example of such process might be the atypical adenomatous hyperplasia in the lung. Destabilization of the equilibrium in such system represents initial invasion of cancer. We are looking for a transition from a slightly perturbed equilibrium state to uncontrolled and irregular growth. We examine a mathematical model of a population of cells distributed over a linear or tubular structure. Growth of cells is regulated by a growth factor, which can diffuse over the structure. Aside from this, production of cells and of the growth factor is governed by a pair of ordinary differential equations. Equation for the cell number follows from an accepted model of cell cycle. Equation for the bounded receptor particle number follows from a time-continuous Markov process. We demonstrate existence of the solutions of the complete model, using the method of invariant rectangles. We find conditions under which diffusion causes destabilization of the spatially homogeneous steady state, leading to exponential growth and apparently chaotic spatial patterns, following a period of almost constancy. This phenomenon may serve as a mathematical explanation of "unexpected" rapid growth and invasion of temporarily stable structures composed of cancer cells.     

Copper deprivation in rats induces islet hyperplasia and hepatic metaplasia in the pancreas

BACKGROUND INFORMATION: Prolonged copper deprivation in rats followed by refeeding with a normal diet has previously been used to induce the appearance of hepatocyte-like cells in the pancreas, but the effects on islet size and morphology have not been determined. RESULTS: In the present study we investigated the distribution of pancreatic alpha- and beta-cells and of hepatocytes in adult rats fed a copper-deficient diet followed by refeeding with a normal diet. Immunohistochemical staining for insulin and glucagon showed that the islets of the copper-deficient group were up to 2.4 times larger in mass compared with controls. The islets were disorganized, with alpha-cells found in multiple layers at the periphery of the islet and sometimes deep in the core. Isolated alpha- and beta-cells were also found in increased numbers in the ductular system. Copper deprivation caused almost complete ablation of the acinar cells, and refeeding induced adipogenesis, acinar regeneration and hepatocyte-like cells. Ductular proliferation and nerve hyperplasia were also present. The hepatocytes tended to be associated with islets or with ducts, rather than with residual pancreatic exocrine tissue. CONCLUSIONS: These data show that copper deficiency in rats, as well as inducing the appearance of hepatocytes, is capable of causing islet hyperplasia.     

采用SM 22启动子调节荧光蛋白的策略分选和鉴定人前列腺平滑肌细胞与成纤维细胞

 平滑肌细胞的数量、表型以及在间质细胞中所占的比例在前列腺间质增生的发生和发展中占有重要的地位.获得纯的平滑肌细胞和成纤维细胞,研究它们基因表达的差异,有助于进一步揭示前列腺增生的分子病因学.构建不同长度的 SM 22启动子,测定荧光素酶活性.采用了基于启动子特异性激活红绿色荧光蛋白表达结合流式细胞分选的策略,体外分离纯的平滑肌细胞和成纤维细胞.启动子活性实验结果表明,1 396 bp的人SM22启动子具有平滑肌细胞特异性和较高的相对活性.构建了红绿荧光蛋白的表达载体pDual-color,在此载体中,RFP的表达受1 396bp的SM22启动子调控,GFP的组成型表达受CMV启动子控制.用流式细胞仪分选GFP+/RFP+和GFP+/RFP-细胞,提取总RNA,进行实时定量RT-PCR.结果显示,在分选获得的GFP+/RFP+细胞比GFP+/RFP-细胞的SM22和SMMHC表达水平高10倍以上.提示,基于启动子特异性可以在体外分离纯的平滑肌细胞和成纤维细胞.