Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential

学习的主要目标是多识别人的新奇来源有势力房间，克服涉及胚胎的干细胞研究和很成年的干细胞的有限可获得性的道德的问题。羊膜的液体房间(声频抗流圈) 习惯性地为出生前的诊断被获得并且能在 vitro 被扩展；他们的起源和性质的不过当前的知识是有限的。声频抗流圈的二十件样品在文化暴露于 adipogenic， osteogenic，神经原并且 myogenic 媒介。区别用免疫细胞化学， RT-PCR 并且西方的弄污被评估。在处理前，声频抗流圈显示出异构的形态学。他们为 MyoD， Myf-5， MRF4， Myogenin 和 Desmin 是否定的但是为由 RT-PCR 的 osteocalcin， PPARgamma2， GAP43， NSE， Nestin， MAP2， GFAP 和贝它导管素 III 积极。房间表示了 Oct-4， Rex-1 和 Runx-1，它描绘无差别的干细胞状态。由免疫细胞化学，他们表示了 neural-glial 蛋白质，间充质、上皮的标记。在文化以后，当占优势的细胞的部件是 fibroblastic 时，声频抗流圈区分了进 adipocytes 和造骨细胞。就算没有 neuronal 形态学是可检测的，早、迟了的 neuronal 抗原仍然是在在神经特定的媒介的 2 星期文化以后的现在。我们的结果提供人的羊膜的液体与为几个系显示出茎和织物特定的基因 / 蛋白质存在的多系潜力包含祖先房间的证据。

Study of fluid cell coarsening for CFD-DEM simulations of polydisperse gas–solid flows

Particle polydispersity is ubiquitous in industrial fluidized beds,which possesses a significant impact on hydrodynamics of gas-solid flow.Computational fluid dynamics-discrete element method(CFD-DEM)is promising to adequately simulate gas-solid flows with continuous particle size distribution(PSD)while it still suffers from high computational cost.Corresponding coarsening models are thereby desired.This work extends the coarse-grid model to polydisperse systems.Well-resolved simulations with different PSDs are processed through a filtering procedure to modify the gas-particle drag force in coarse-grid simulations.We reveal that the drag correction of individual particle exhibits a dependence on filtered solid volume fraction and filtered slip velocity for both monodisperse and polydisperse systems.Subsequently,the effect of particle size and surrounding PSD is quantified by the ratio of particle size to Sauter mean diameter.Drag correction models for systems with monodisperse and continuous PSD are developed.A priori analysis demonstrates that the developed models exhibit reliable prediction accuracy.

Heart Valves Grown from Womb Fluid Cells

科学家们首次从人体子宫的流体细胞中培育出了人类的心瓣膜,这一发现为有先天心脏缺陷的患者带来了福音。这种由自身组织培植的心瓣膜被移植入人体后,其性能将比人工心脏或者通过器官捐献而来的心脏更持久有效;并且,此项研究的前期动物试验已经成功。

Osteogenic differentiation of amniotic fluid mesenchymal stromal cells and their bone regeneration potential

In orthopedics, tissue engineering approach using stem cells is a valid line of treatment for patients with bone defects. In this context, mesenchymal stromal cells of various origins have been extensively studied and continue to be a matter of debate. Although mesenchymal stromal cells from bone marrow are already clinically applied, recent evidence suggests that one may use mesenchymal stromal cells from extra-embryonic tissues, such as amniotic fluid, as an innovative andadvantageous resource for bone regeneration. The use of cells from amniotic fluid does not raise ethical problems and provides a sufficient number of cells without invasive procedures. Furthermore, they do not develop into teratomas when transplanted, a consequence observed with pluripotent stem cells. In addition, their multipotent differentiation ability, low immunogenicity, and anti-inflammatory properties make them ideal candidates for bone regenerative medicine. We here present an overview of the features of amniotic fluid mesenchymal stromal cells and their potential in the osteogenic differentiation process. We have examined the papers actually available on this regard, with particular interest in the strategies applied to improve in vitro osteogenesis. Importantly, a detailed understanding of the behavior of amniotic fluid mesenchymal stromal cells and their osteogenic ability is desirable considering a feasible application in bone regenerative medicine.

Cerebrospinal fluid and neural stem cell niche control

Neurogenesis from inner brain neural stem cells （NSCs） is a process which takes place continuously in mammals through- out their life. However, the main ontogenic difference is the intensity of neurogenesis, which commences as a very intensive and global activity in the early embryonic brain （neural tube）, persists in fetal and newborn stages, and declines significantly in adulthood, becoming restricted to specific places with low neurogenic activity such as the subventricular zone （SVZ） and the subgranular zone （SGZ） in the dentate gyrus （DG） of the hippocampus.

Neural differentiation of choroid plexus epithelial cells:role of human traumatic cerebrospinal fluid

As the key producer of cerebrospinal fluid（CSF）,the choroid plexus（CP） provides a unique protective system in the central nervous system.CSF components are not invariable and they can change based on the pathological conditions of the central nervous system.The purpose of the present study was to assess the effects of non-traumatic and traumatic CSF on the differentiation of multipotent stem-like cells of CP into the neural and/or glial cells.CP epithelial cells were isolated from adult male rats and treated with human non-traumatic and traumatic CSF.Alterations in m RNA expression of Nestin and microtubule-associated protein（MAP2）,as the specific markers of neurogenesis,and astrocyte marker glial fibrillary acidic protein（GFAP） in cultured CP epithelial cells were evaluated using quantitative real-time PCR.The data revealed that treatment with CSF（non-traumatic and traumatic） led to increase in m RNA expression levels of MAP2 and GFAP.Moreover,the expression of Nestin decreased in CP epithelial cells treated with non-traumatic CSF,while treatment with traumatic CSF significantly increased its m RNA level compared to the cells cultured only in DMEM/F12 as control.It seems that CP epithelial cells contain multipotent stem-like cells which are inducible under pathological conditions including exposure to traumatic CSF because of its compositions.

Computational fluid dynamics simulation of gas-liquid two phases flow in 320 m^3 air-blowing mechanical flotation cell using different turbulence models

According to the recently developed single-trough floating machine with the world's largest volume(inflatable mechanical agitation flotation machine with volume of 320 m3) in China, the gas-fluid two-phase flow in flotation cell was simulated using computational fluid dynamics method. It is shown that hexahedral mesh scheme is more suitable for the complex structure of the flotation cell than tetrahedral mesh scheme, and a mesh quality ranging from 0.7 to 1.0 is obtained. Comparative studies of the standard k-ε, k-ω and realizable k-ε turbulence models were carried out. It is indicated that the standard k-ε turbulence model could give a result relatively close to the practice and the liquid phase flow field is well characterized. In addition, two obvious recirculation zones are formed in the mixing zones, and the pressure on the rotor and stator is well characterized. Furthermore, the simulation results using improved standard k-ε turbulence model show that surface tension coefficient of 0.072, drag model of Grace and coefficient of 4, and lift coefficient of 0.001 can be achieved. The research results suggest that gas-fluid two-phase flow in large flotation cell can be well simulated using computational fluid dynamics method.

Deleterious effects of postburn subeschar tissue fluid on cultured cells

Objective: To investigate the role of subeschar tissue fluid (STF) in postburn cell damage. Methods: STF was collected from patients with full-thickness burn injury of 10% - 69% total body surface area (TFBSA) during operation on postburn day 2 - 4, and that from guinea pigs were obtained in escharectomy 24 h after full-thickness scald injury of 30%’TBSA was induced. Respective incubation of human umbilical vein endothelial cells (HUVEC) and hepatocytes with STF were performed for the observation of the effects of STF on the activity, morphology, physiological function and monolayer cell permeability of HUVEC, and the percentages of apoptotic and necrotic hepatocytes were determined. Results: (l) STF had evident deleterious effect on endothelial cells. (2)are could induce apoptosis and necrosis of hepatocytes. Conclusion: STF plays an important role in causing early postburn damages.

Comparison of human amniotic fluid-derived and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells： Characterization and myocardial differentiation capacity

ObjectiveTo 比较描述和羊膜的导出液体的间充质的 stromal 房间(AF MSC ) 和脐的绳索沃顿 .MethodsThe 人 AF MSC 是的导出果冻的间充质的 stromal 房间(WJ MSC ) 的心肌的区别能力从羊水诊断获得的羊膜的液体样品有教养。脐的绳索 WJ MSC 从沃顿由正常劳动完整术语的交付的婴儿的脐的绳索的果冻被获得。由房间表面标记的流动 cytometry 的形态学，生长曲线，和分析在房间的二种类型之间被比较。心肌的基因(GATA-4， c-TnT， &#x003b1； -actin, 和 Cx43 ) 被即时 PCR 检测，相应蛋白质表情被西方的污点分析检测在以后心肌在 AF MSC 导致了， WJ MSCs.ResultsOur 调查结果表明 AF MSC 和 WJ MSC 分享了 fibroblastoid 形状的类似的词法特征。AF MSC 容易比 WJ MSC 被获得并且有一更短的时间到达 2.7 &#x000b1 的坚持；1.6 天到 6.5 &#x000b1 的 WJ MSC；1.8 天。由 MTT 细胞毒素的试金的生长曲线证明 AF MSC 在经过 5 和经过 10 点有一个类似的 proliferative 能力。然而， WJ MSC 的 proliferative 能力相对 10 经过在 5 经过被减少。AF 干细胞和 WJ 干细胞与胚胎的干细胞的一些特征有间充质的 stromal 房间的特征。他们表示 CD29 和 CD105，然而并非 CD34。他们为一级主要 histocompatibility 是积极的(MHC 我) 抗原(HLA-ABC ) ，并且否定，或略微积极，为 MHC 班 II (HLA 医生) 抗原。Oct-4 在所有二种房间类型是积极的。AF MSC 和 WJ MSC 能区分心肌层。区别能力被 GATA-4 的表达式检测， c-TnT， &#x003b1；在心肌的 induction.ConclusionsBoth AF MSC 以后的 -actin, Cx43 和 WJ MSC 在心脏的再生治疗为 myogenesis 有潜在的临床的应用程序。

Cerebrospinal fluid used as culture medium prior to autologous olfactory ensheathing cell transplantation

BACKGROUND： Cerebrospinal fluid can be an inducer for neural stem cells in vitro, but few studies employ cerebrospinal fluid to culture olfactory ensheathing cells. OBJECTIVE： To investigate the growth of nasal mucosa olfactory ensheathing cells in normal cerebrospinal fluid, and to analyze the feasibility of cerebrospinal fluid for culturing olfactory ensheathing cells used for transplantation. DESIGN, TIME AND SETTING： A completely randomized, block design study was performed at the Cell Laboratory, Wuxi Third People＇s Hospital, and Jiangsu Institute of Parasitic Diseases, China, in August 2008. MATERIALS： Dulbecco＇s modified Eagle＇s medium/F12 （DMEM/F12） and fetal bovine serum （Gibco BRL, USA）, mouse anti-rat P75 monoclonal antibody and rabbit anti-glial fibrillary acidic protein polyclonal anti body （＇Santa Cruz Biotechnology, USA）, mouse anti-rat myelin basic protein monoclonal antibody （Cymbus, UK）, mouse anti-rat microtubule-associated protein-2 monoclonal antibody （Transduction Laboratories, USA）, FITC conjugated rabbit anti-mouse monoclonal antibody （Boster, China）, TRITC conjugated goat anti-rabbit monoclonal antibody （Sigma, USA） were used. METHODS： Nasal mucosa olfactory ensheathing cells were separately incubated in DMEM/F12, cerebrospinal fluid, and changing DMEM/F12 into cerebrospinal fluid. Adult female Sprague Dawley rat models of spinal hemisection were established. Nerve injury was repaired by transplantation of nasal mucosa olfactory ensheathing cells cultured in cerebrospinal fluid or DMEM/F12. MAIN OUTCOME MEASURES： The proliferative ability of olfactory ensheathing cells cultured in cerebrospinal fluid was determined by a 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium bromide assay. The morphology and purity of olfactory ensheathing cells were detected using immunohistochemistry. Animal behavior was evaluated by the Basso, Beattie and Bresnahan locomotor rating scale. Morphological repair was assessed by a horseradish peroxidase-tetramethylbenzidine retrograde tracer technique and immunohistochemistry. RESULTS： Changing from DMEM/F12 to cerebrospinal fluid did not change overall culture morphology and purity on day 14. These cells also contributed to myelinization and the conduction velocity of regenerated axons, and improved motor abilities of denervated muscle fibers in rats with spinal cord injury. The recovery of behavioral function and neuronal regeneration was similar in the two groups. CONCLUSION： Cerebrospinal fluid culture prior to autologous olfactory ensheathing cell transplantation is feasible for clinical use.

Senescence and longevity in amniotic fluid derived cells

Amniotic fluid stem cells(AFSC) are noncontroversial and abundant potentially transplantable cells. They are derived from fetal and amniotic sources and show osteogenic, neuronal, and adipogenic differentiation in culture. Clinical material must be expanded in culture and sorted if clinical use is desired. Cellular senescence and replicative potential for AFSC cultures has had limited study, with scant data on gene expression over time. We report changes in samples from 17 patients over multiple passages form 10 to 81 population doublings. Longevity was unrelated to telomere length in these cells. It was related to upregulation of TWIST1, which is highly expressed in stem cells, and downregulation of genes associated with apoptosis.

Amniotic fluid stem cell-based models to study the effects of gene mutations and toxicants on male germ cell formation

Male infertility is a major public health issue predominantly caused by defects in germ cell development. In the past, studies on the genetic regulation of spermatogenesis as well as on negative environmental impacts have been hampered by the fact that human germ cell development is intractable to direct analysis in vivo. Compared with model organisms including mice, there are fundamental differences in the molecular processes of human germ cell development. Therefore, an in vitro model mimicking human sperm formation would be an extremely valuable research tool. In the recent past, both human embryonic stem （ES） cells and induced pluripotent stem （iPS） cells have been reported to harbour the potential to differentiate into primordial germ cells and gametes. We here discuss the possibility to use human amniotic fluid stem （AFS） ceils as a biological model. Since their discovery in 2003, AFS cells have been characterized to differentiate into cells of all three germ layers, to be genomically stable, to have a high proliferative potential and to be non-tumourigenic. In addition, AFS cells are not subject of ethical concerns. In contrast to iPS cells, AFSs cells do not need ectopic induction of pluripotency, which is often associated with only imperfectly cleared epigenetic memory of the source cells. Since AFS cells can be derived from amniocentesis with disease-causing mutations and can be transfected with high efficiency, they could be used in probing gene functions for spermatogenesis and in screening for male reproductive toxicity.

Identification of interleukin-6 (IL-6) and squamous cell carcinoma (SCC) as amniotic fluid-specific markers

Objective: To determine if an amniotic fluid (AF)-specific marker is present and if its concentration changes with the presence of labor. Study Design: Twenty-six healthy women who gave birth to healthy newborns at term during the period from July 2009 to January 2010 were included in the study. Six candidate markers were assessed by commercially available ELISA kits: interleukin (IL)-6, squamous cell carcinoma (SCC) antigen, insulin-like growth factor (IGFBP)-1, osteopontin (OPN), CA125, and sialyl Tn (STN). Results: The AF/maternal serum (MS) measurement based on IL-6 or SCC has proved to be superior to IGFBP-1, CA125, OPN and STN. Women with spontaneous labor at term had significantly higher IL-6 and IGFBP-1 concentrations in AF compared with those without labor. No significant differences were observed in the AF concentrations of SCC, OPN, CA125 and STN between women with labor and those not in labor. Conclusion: Our observation of IL-6 and SCC in AF may open a new area of research to assess their usefulness as biological markers of obstetrical disorders.

Optimization study of a PEM fuel cell performance using 3D multi-phase computational fluid dynamics model

An optimization study using a comprehensive 3D, multi-phase, non-isothermal model of a PEM (proton exchange membrane) fuel cell that incorporates significant physical processes and key parameters affecting fuel cell performance is pre-sented and discussed in detail. The model accounts for both gas and liquid phase in the same computational domain, and thus allows for the implementation of phase change inside the gas diffusion layers. The model includes the transport of gaseous species, liquid water, protons, energy, and water dissolved in the ion-conducting polymer. Water is assumed to be exchanged among three phases: liquid, vapour, and dissolved, with equilibrium among these phases being assumed. This model also takes into account convection and diffusion of different species in the channels as well as in the porous gas diffusion layer, heat transfer in the solids as well as in the gases, and electrochemical reactions. The results showed that the present multi-phase model is capable of iden-tifying important parameters for the wetting behaviour of the gas diffusion layers and can be used to identify conditions that might lead to the onset of pore plugging, which has a detrimental effect on the fuel cell performance. This model is used to study the effects of several operating, design, and material parameters on fuel cell performance. Detailed analyses of the fuel cell per-formance under various operating conditions have been conducted and examined.

Numerical simulation of a direct internal reforming solid oxide fuel cell using computational fluid dynamics method

A detailed mathematical model of a direct internal reforming solid oxide fuel cell(DIR-SOFC) incorporating with simulation of chemical and physical processes in the fuel cell is presented. The model is developed based on the reforming and electrochemical reaction mechanisms,mass and energy conservation,and heat transfer. A computational fluid dynamics(CFD) method is used for solving the complicated multiple partial differential equations(PDEs) to obtain the numerical approximations. The resulting distributions of chemical species concentrations,temperature and current density in a cross-flow DIR-SOFC are given and analyzed in detail. Further,the influence between distributions of chemical species concentrations,temperature and current density during the simulation is illustrated and discussed. The heat and mass transfer,and the kinetics of reforming and electrochemical reactions have significant effects on the parameter distributions within the cell. The results show the particular characteristics of the DIR-SOFC among fuel cells,and can aid in stack design and control.

肺癌病理应用支气管冲洗细胞块免疫组化链霉菌亲生物素蛋白-过氧化物酶连接法染色的诊断效果及检出率评价

目的探讨肺癌病理应用支气管冲洗细胞块免疫组化链霉菌亲生物素蛋白-过氧化物酶连接法(SP)染色的诊断效果及检出率。方法选取2017年2月至2021年12月广东省开平市中心医院收治的70例经支气管冲洗液检查阳性患者作为研究对象。采用液基薄层细胞学(TCT)技术制备细胞块,分别采用免疫组化SP法、苏木精伊红染色(HE)进行常规细胞学检查,比较两种检测方法诊断效能及不同肺癌类型免疫组化SP法检查后不同抗体阳性表达率比较。结果免疫组化SP法对恶性肿瘤诊断的灵敏度为96.15%、准确度为95.71%,均高于HE染色的69.23%、70.00%(P<0.05),特异度为94.44%,高于HE染色的72.22%,但差异无统计学意义。免疫组化SP法对肺癌病理类型总检出率高于HE染色,差异有统计学意义(P<0.05)。免疫组化SP法对肌上皮(p63)、细胞角蛋白5/6抗体(CK5/6)、细胞角蛋白7(CK7);甲状腺转录因子-1(TTF-1)、突触素(Syn)、人表皮生长因子受体-5(CD56),增殖细胞的核抗原(Ki67),天冬氨酸蛋白酶(Napsin-A)表达阳性率均高于HE染色,差异统计学意义(P<0.05)。不同类型肺癌患者p63、CK5/6、CK7、TTF-1、Syn、CD56、Ki67、Napsin-A阳性表达率比较差异有统计学意义(P<0.05)。结论支气管冲洗液细胞块免疫组化SP法染色对肺癌有较高的诊断价值,对不同类型肺癌有较高的检出率及鉴别率,具有重要的临床意义。

脑脊液免疫球蛋白、外周血炎症细胞指标及细胞免疫指标在儿童自身免疫性脑炎中的临床诊断价值

目的探讨脑脊液免疫球蛋白、外周血炎症细胞指标及细胞免疫指标在儿童自身免疫性脑炎(AE)中的临床诊断价值。方法选取2022年1月至2023年12月江西省儿童医院收治的36例AE患儿纳入AE组,另选取江西省儿童医院同期收治的36例病毒性脑炎(VE)儿童作为VE组。比较两组的脑脊液IgM、IgG、IgA水平和外周血中性粒细胞计数/淋巴细胞计数比值(NLR)、单核细胞计数/淋巴细胞计数比值(MLR)、血小板计数/淋巴细胞计数(PLR)、CD3^(+)、CD4^(+)/CD8^(+)、自然杀伤(NK)细胞比例;采用受试者工作特征(ROC)曲线分析脑脊液免疫球蛋白、外周血炎症细胞指标及细胞免疫指标对AE的诊断价值。结果AE组的脑脊液IgM、IgG和IgA水平高于VE组,差异有统计学意义(P<0.05);AE组的NLR、MLR和PLR高于VE组,差异有统计学意义(P<0.05);AE组的CD3^(+)、CD4^(+)/CD8^(+)和NK细胞比例水平低于VE组,差异有统计学意义(P<0.05);ROC曲线分析结果显示,IgM、IgG、IgA、NLR、MLR、PLR、CD3^(+)、CD4^(+)/CD8^(+)、NK细胞比例诊断AE的曲线下面积(AUC)分别为0.844、0.874、0.885、0.741、0.767、0.668、0.793、0.796和0.671(P<0.05)。结论脑脊液免疫球蛋白、外周血炎症细胞指标及细胞免疫指标对AE均具有一定的诊断价值,其中脑脊液免疫球蛋白水平诊断效能最高。

Coscinium fenestratum: Callus and Suspension Cell Culture of the Endangered Medicinal Plant Using Vermicompost Extract and Coelomic Fluid as Plant Tissue Culture Media

In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were induced to form callus when cultured on vermicompost extract media along with coelomic fluid. Suspension medium was developed using vermicompost extract and coelomic fluid in 3:1 ratio. Phytochemical analysis of the alkaloid berberine was confirmed from callus, suspension cell culture and suspension medium by Thin Layer Chromatography and High Performance Liquid Chromatography. Vermicompost and its extracts with coelomic fluid have shown maximum (100 per cent) response of callus induction. Callus mass enlarged with increasing concentration of coelomic fluid and callus growth was assessed from the biomass. Incubation of culture tubes in dark supported callus development significantly. The Rf value of 0.36 confirmed the presence of berberine by Thin Layer Chromatography. Qualitative analysis confirmed the presence of alkaloid berberine with the retention time of 2.8 minutes similar to that of standard reference sample from Sigma chemicals, USA. The suspension medium turned deep yellow because of the release of the alkaloid. Vermicompost and its extracts along with coelomic fluid have shown the economical approach for micropropagation of economically and medicinally important plants.

解毒抑瘤汤联合西医标准方案治疗基因突变阴性晚期非小细胞肺癌的疗效观察

目的:观察基于“癌毒-气血津液”病机理论组方的解毒抑瘤汤联合西医标准方案治疗基因突变阴性晚期非小细胞肺癌(Non-small cell lung cancer,NSCLC)的疗效及毒副反应。方法:收集基因突变阴性晚期NSCLC病例共48例,采用随机数字表法分为治疗组与对照组,每组各24例,对照组采用西医标准方案治疗,治疗组在对照组基础上给予中药汤剂解毒抑瘤汤加减,以21天为1个周期,连续治疗2个周期后评价两组患者的近期疗效、中医证候疗效、生活质量、中位无进展生存期及毒副反应。结果:治疗组vs对照组:治疗有效率:54.2%vs 45.8%;疾病控制率:83.3%vs 70.8%;中医证候疗效:87.5%vs 70.8%;KPS评分:78.49±12.81分vs 69.32±13.94分;中位无进展生存期:9.8月vs 6.2月。治疗组在治疗有效率、疾病控制率、中医证候疗效、KPS评分及中位无进展生存期方面均优于对照组(P<0.05)。治疗组的恶心呕吐、便秘、纳差乏力发生率低于对照组(P<0.05),治疗组的骨髓抑制和肝肾损害发生率与对照组相比无统计学差异(P>0.05)。结论:解毒抑瘤汤能提高西医标准方案治疗基因突变阴性晚期NSCLC的近期疗效,延长患者生存时间,改善患者中医证候和生活质量,同时能减轻化疗相关部分副反应,且安全性好。