In vitro derivation of functional insulin-producing cells from human embryonic stem cells

The capacity for self-renewal and differentiation of human embryonic stem （ES） cells makes them a potential source for generation of pancreatic beta cells for treating type I diabetes mellitus. Here, we report a newly developed and effective method, carried out in a serum-free system, which induced human ES cells to differentiate into insulin-producing cells. Activin A was used in the initial stage to induce definitive endoderm differentiation from human ES cells, as detected by the expression of the definitive endoderm markers Sox17 and Brachyury. Further, all-trans retinoic acid （RA） was used to promote pancreatic differentiation, as indicated by the expression of the early pancreatic transcription factors pdxl and hlxb9. After maturation in DMEM/F12 serum-free medium with bFGF and nicotinamide, the differentiated cells expressed islet specific markers such as C-peptide, insulin, glucagon and glut2. The percentage of C-peptide-positive cells exceeded 15%. The secretion of insulin and C-peptide by these cells corresponded to the variations in glucose levels. When transplanted into renal capsules of Streptozotocin （STZ）-treated nude mice, these differentiated human ES cells survived and maintained the expression of beta cell marker genes, including C-peptide, pdxl, glucokinase, nkx6.1, lAPP, pax6 and Tcfl. Thirty percent of the transplanted nude mice exhibited apparent restoration of stable euglycemia; and the corrected phenotype was sustained for more than six weeks. Our new method provides a promising in vitro differentiation model for studying the mechanisms of human pancreas development and illustrates the potential of using human ES cells for the treatment of type I diabetes mellitus.

Highly efficient differentiation of human ES cells and iPS cells into mature pancreatic insulin-producing cells

Human pluripotent stem cells represent a potentially unlimited source of functional pancreatic endocrine lineage cells. Here we report a highly efficient approach to induce human embryonic stem （ES） cells and induced pluripo- tent stem （iPS） cells to differentiate into mature insulin-producing cells in a chemical-defined culture system. The differentiated human ES cells obtained by this approach comprised nearly 25% insulin-positive cells as assayed by flow cytometry analysis, which released insulin/C-peptide in response to glucose stimuli in a manner comparable to that of adult human islets. Most of these insulin-producing cells co-expressed mature β cell-specific markers such as NKX6-1 and PDX1, indicating a similar gene expression pattern to adult islet β cells in vivo. In this study, we also demonstrated that EGF facilitates the expansion of PDXl-positive pancreatic progenitors. Moreover, our protocol also succeeded in efficiently inducing human iPS cells to differentiate into insuIin-producing ceils. Therefore, this work not only provides a new model to study the mechanism of human pancreatic specialization and maturation in vitro, but also enhances the possibility of utilizing patient-specific iPS cells for the treatment of diabetes.

A secretory function of human insulin-producing cells in vivo

BACKGROUND: Mesenchymal stem cells derived from human umbilical cord blood (UCB-MSCs) have good research and application prospects in the treatment of diabetes. We once induced UCB-MSCs to differentiate into insulin-producing cells (IPCs) in vitro, but we did not know the functions of these cells in vivo. The aim of this study was to assess the functional effects of IPCs on insulin secretion and their role in the treatment of diabetes in vivo. METHODS: UCB-MSCs were induced to IPCs by an inducing protocol with extracellular matrix gel. BALB/C nude mice were made hyperglycemic by intraperitoneal injection of streptozotocin. The diabetic mice were transplanted with 1x10(7) IPCs under the renal capsule or with phosphate-buffered saline as a control. After transplantation, the grafts were analyzed by immunocytochemistry for the expression of human insulin; the serum human insulin levels were measured; and blood glucose and body weight status were monitored. RESULTS: Immunofluorescence showed that numerous IPCs under the kidney capsule were insulin-positive. On day 14 after transplantation, the serum human insulin level of the treatment group (n=9) averaged 0.44 +/- 0.12 mU/L, which was higher than that of the control group (n=9) that did not express insulin (t=10.842, P<0.05). The diabetic mice remained hyperglycemic and kept losing body weight after IPC transplantation, and there was no significant difference in the control group. CONCLUSION: IPCs differentiated from UCB-MSCs generate human insulin in diabetic mice, but more research is needed to make further use of them to regulate hyperglycemia and body weight in vivo. (Hepatobiliary Pancreat Dis Int 2009; 8: 255-260)

In vitro-derived insulin-producing cells modulate Th1 immune responses and induce IL-10 in streptozotocin-induced mouse model of pancreatic insulitis

Background: Insulitis is defined by the presence of immune cells infiltrating in the pancreatic islets that might progress into the complete β-cell loss. The immunomodulatory properties of bone marrow-derived mesenchymal stem cells(BM-MSCs) have attracted much attention. This study aimed to evaluate the possible immunomodulatory effects of rat BM-MSCs and MSCs-derived insulin-producing cells(IPCs) in a mouse model of pancreatic insulitis. Methods: Insulitis was induced in BALB/c mice using five consecuti ve doses of streptozotocin. MSCs or IPCs were directly injected into the pancreas of mice and their effects on the expression of Th subsetsrelated genes were evaluated. Results: Both BM-MSCs and IPCs significantly reduced the expression of pancreatic Th1-related IFN-γ( P < 0.001 and P < 0.05, respectively) and T-bet genes(both P < 0.001). Moreover, the expression of IL-10 gene was significantly increased in IPC-treated compared to BM-MSC-or PBS-treated mice( P < 0.001 both comparisons). Conclusions: BM-MSCs and IPCs could successfully suppress pathologic Th1 immune responses in the mouse model of insulitis. However, the marked increase in IL-10 gene expression by IPCs compared to BM-MSCs suggests that their simultaneous use at the initial phase of autoimmune diabetes might be a better option to reduce inflammation but these results need to be verified by further experiments.

Induction of pancreatic duct cells of neonatal rats into insulin-producing cells with fetal bovine serum: A natural protocol and its use for patch clamp experiments

AIM： To induce the pancreatic duct cells into endocrine cells with a new natural protocol for electrophysiological study. METHODS： The pancreatic duct cells of neonatal rats were isolated, cultured and induced into endocrine ceils with 15% fetal bovine serum for a period of 20 d. During this period, insulin secretion, MTT value, and morphological change of neonatal and adult pancreatic islet cells were comparatively investigated. Pancreatic β-cells were identified by morphological and electrophysiological characteristics, while ATP sensitive potassium channels （KATP）, voltage-dependent potassium channels （Kv）, and voltage-dependent calcium channels （KcA） in β-cells were identified by patch clamp technique. RESULTS： After incubation with fetal bovine serum, the neonatal duct cells budded out, changed from duct-like cells into islet clusters. In the first 4 d, MTT value and insulin secretion increased slowly （MTT value from 0.024 ±0.003 to 3.028±0.003, insulin secretion from 2.6±0.6 to 3.1±0.8 mIU/L）. Then MTT value and insulin secretion increased quickly from d 5 to d 10 （MTT value from 0.028 ±0.003 to 0.052±0.008, insulin secretion from 3.1±0.8 to 18.3±2.6 mIU/L）, then reached high plateau （MTT value 〉0.052±0.008, insulin secretion 〉18.3±2.6 mIU/L）. In contrast, for the isolated adult pancreatic islet cells, both insulin release and MTT value were stable in the first 4 d （MTT value from 0.029±0.01 to 0.031±0.011, insulin secretion from 13.9±3.1 to 14.3±3.3 mIU/L）, but afterwards they reduced gradually （MTT value 〈0.031 ±0.011, insulin secretion 〈8.2±1.5 mIU/L）, and the pancreatic islet cells became dispersed, broken or atrophied correspondingly. The differentiated neonatal cells were identified as pancreatic islet cells by dithizone staining method, and pancreatic β-cells were further identified by both morphological features and electrophysiological characteristics, i.e. the existence of recording currents from KATP, Kv, and KCA. CONCLUSION： Islet cells differentiated from neonatal pancreatic duct cells with the new natural protocol are more advantageous in performing patch clamp study over the isolated adult pancreatic islet cells.

未来5~10年中国医疗保障待遇设计的改革思路

医疗保障待遇包括起付线、报销比例、封顶线等内容。典型国家社会医疗保险报销起付线多设置在人均年收入的3%以内,对门诊、住院和药剂服务设立了不同的支付政策,医疗费用个人自付占可支配收入的比例在0.25%~2%。借鉴国际经验,展望中国社会医疗保险制度未来的发展,建议提高门诊保障待遇,以可支配收入为基数设置报销起付线,不同层级医疗机构实行相同的报销比例,设置个人自付封顶线,取消门诊报销封顶线,门诊和住院共用个人最高自付限额。

2023年积石山地震重要公共建筑吊顶震害调查分析

对2023年6.2级积石山地震中公共建筑内部吊顶开展了震害调查及分析,并以积石山县大河家镇大河家中学教学楼、中西医结合医院门诊楼以及中国邮政银行会议室3处典型震害为例,分析了在以往的公共建筑吊顶震害调查及研究中较少涉及的铝扣板吊顶、间接悬挂式吊顶的新震害现象及典型震害特征。研究表明:铝扣板边角锋利,卡入式连接对龙骨网格平整度要求较高,此类吊顶如若用于抗震要求较高或人员较为密集的建筑内,建议进行专门的抗震分析,以避免坠板伤人或妨碍人员逃生。此外,在吊顶地震损伤评估中,除坠板率外,还应考虑龙骨网格变形等因素。应加强吊顶中照明单元与主体结构连接的可靠性以避免其在地震中出现大量坠落。研究结果可为公共建筑吊顶抗震研究与工程应用提供参考。

风扇与辐射空调耦合系统的热舒适分析

针对顶板辐射供冷系统存在的噪声问题,提出了一种采用辐射供冷与吸音板耦合系统,同时,通过在辐射顶板和吸音板之间增加风扇(向下吹),来抵消由于吸音板导致的辐射热交换的减少。重点研究了顶板辐射供冷与风扇耦合系统的室内气流分布、热舒适以及传热系数变化,其影响因素包括:吸音板覆盖率、风扇转速、房间高度。结果表明:风扇的加入增强了顶板的对流换热和总换热;随着风扇转速的增加和吸音板覆盖率的降低,室内空气温度降低;与无风扇的顶板辐射系统相比,风扇在高转速工况下的PMV值降低了0.37~0.40,总传热系数增长了28.4%~45.4%,在吸音板覆盖率为16%时,总传热系数的增长率达到峰值;可以通过增加风扇从而强化室内空气流动的方式来补偿较高室温带来的不适感。研究结果为顶板辐射系统的实施提供了有前景和实用的设计方案。

工程车辆直线电机馈能悬架能量回收分析与参数优化

目的为了回收工程车辆在行驶过程中由于路面不平度引起的车身振动能量,同时解决馈能悬架系统的动力学性能和馈能性能不协调问题,方法首先,提出一种直线电机与减振器并联的馈能悬架系统结构,通过建立馈能悬架系统的二自由度动力学模型,利用天棚控制提高车辆的平顺性;其次,以悬架减振器阻尼作为优化设计变量,结合直线电机式馈能悬架的馈能性能、平顺性和安全性建立优化目标函数,采用遗传算法控制策略对馈能悬架的馈能性能进行优化,以获得全局最优解,解决馈能悬架的平顺性和馈能性能的协调问题;最后,通过MATLAB/Simulink对馈能悬架系统性能进行仿真。结果仿真结果表明:与被动悬架相比,天棚控制下的馈能悬架车身加速度、轮胎动载荷、悬架动行程均方根值分别减小25.40%,17.76%和32.58%;利用遗传算法对馈能悬架的动力性能和馈能协调性能优化,优化后馈能悬架的动力学性能相比优化前有一定恶化,但与被动悬架的动力学性能相比提高显著,而蓄电池平均回收能量提高了15.22%,馈能性能提高显著。结论本文提出的直线电机和减振器并联结构的馈能悬架系统装置能够回收工程车辆悬架系统的振动能量,采用遗传算法协调控制策略能够兼顾馈能悬架的动力学性能和馈能性能,并使馈能性能得到显著提高。

基于CFD的辐射冷顶板与风扇耦合系统性能优化

吸音板用来解决辐射式天花板供冷系统的噪声问题,但其阻碍了辐射换热。本文提出通过风扇的调风作用来优化辐射冷顶板与风扇耦合系统,基于CFD模拟讨论了耦合系统的室内气流分布、热舒适及换热系数变化,研究了吸音板安装方式、风扇转速和风扇转向对辐射天花板性能的影响。结果表明:风扇的加入增大了顶板的对流换热系数和总换热系数;随着风扇转速的增大,室内的空气温度和人体热舒适性评价指标(PMV)降低;风扇向上吹风的效果优于向下吹风,吸音板竖直安装优于水平安装。

自然通风条件下隧道火灾动态监测仿真分析

为了降低隧道火灾事故影响,如何动态监测隧道火灾的情况是需要解决的关键性问题。通过FDS场模拟软件建立隧道火灾模型,模拟小型货车车厢位于隧道中部着火的隧道火灾情况。分析火源近场的火源功率及隧道顶部温度变化,探测温度场分布动态变化相关性规律。结果表明:火源功率前200 s内处于稳定阶段,1000 s左右达到最大火源功率。隧道顶部温度整体趋势先升后降,热烟气呈现加速状态,温度变化曲线缓步上升。火源近场隧道顶部温度变化曲线与热释放速率曲线趋于一致,可根据隧道顶部温度变化曲线监测近场火源的动态变化。近场4处测点在12 min内处于临界高程和温度之内,此时火灾扑救人员相对安全,灭火可在此范围内采取内攻方式,为消防救援提供一定参考。

纵向通风作用下公路隧道火灾烟气输运规律研究

在我国交通发展过程中,公路隧道的地位越发显著,隧道火灾危害严重,特别是烟气运动情况对隧道安全至关重要.采用小尺寸模型试验和FDS数值模拟相结合的方法,模拟隧道内火灾发展和烟气输运情况,分析了火场中的火焰情况和烟气运动、温度等参数变化.研究结果表明:当隧道上游入口处的纵向风压由限制风速所需风压增大至临界风速所需风压后,逆流层逐渐消失,最终达到稳态流动模式时,烟气仅从火源的下游开口排出;火源上游的顶棚射流温度衰减要比下游快很多;为保证紧急情况下人员安全疏散,建议隧道火灾早期隧道内的纵向风速宜控制在1.4~1.9m/s.

女性中层管理者职业继续发展阻碍因素的探索研究

为定位“玻璃天花板”的最关键因素及特殊阶段呈现的重要因素,文章聚焦“分水岭”阶段的女性中层管理者群体,构建女性职业发展阻碍因素的理论模型。结果表明,影响女性中层管理者职业继续发展的因素及作用机制可以结合能力—动机—机会(Ability-Motivation-Opportunity,AMO)框架加以理解。组织结构同质化、结构性晋升阻碍和职场性别劣势限制了女性中层管理者职业继续发展机会;社会性别分工认同、工作-家庭平衡成功观,以及低职场性别效能削弱了女性中层管理者职业继续发展的动机;由于工作压力和社会性别评价压力,以及职场任务导向认知、工具性内隐领导原型和性别特质的影响,许多女性中层管理者呈现出侧重任务导向,但任务导向执行低效的领导行为特点,降低了女性中层管理者的领导力,进而制约了女性中层管理者的职业继续发展能力。

理想图景下的顶棚设备处理

在机器美学和诚实信条的影响下,平滑表面构成了现代建筑早期顶棚面的主流理想图景,其意义、实现的形式和方式都随着时代发展而不断变化。顶棚设备系统末端从最初位于顶棚面的下方,发展到早期吊顶时的平齐,再到当代的其后。吊顶面在当代变得独立的同时也获得了实用性之外的美学意义,吊顶空间所整合的环境调控技术提供了呼应外部环境的可能性。技术由外显至内隐的表现变化,使得环境因素对建筑的影响可以超越单纯的形式。

一个包含Smarandache Ceil函数的对偶函数及Euler函数的方程及其可解性

对于给定的自然数n,k,且k≥2,Smarandache Ceil函数的对偶函数珔Sk(n)定义为珔Sk(n)=max{x:x∈N:xk|n}。文中基于∑ d|n Sk(d)=φ(n)的可乘性并运用初等方法分类讨论研究了方程∑d|n Sk(d)=φ(n)的可解性,证明了该方程有仅有限个正整数解,并给出了该方程的所有正整数解。

关于Smarandache ceil函数及其对偶函数的均值

用解析的方法来研究k阶Smarandacheceil函数及其对偶函数与k次幂补数的均值分布性质,并得出几个较为精确的渐近公式.

建筑设计生成方法教学研究——“ceiling Margin”生成工具工程实践

计算机生成方法通过相关算法对建筑元素自组织优化组合,借助传统设计方法不易获得的艺术灵感,它将生成系统作为全新建筑设计方法并趋向艺术实践的程序创作系统。介绍了东南大学2007毕业班建筑生成设计研究组的教学探索及工程实践,通过"ceiling Margin"吊顶生成工具的程序开发、加工及安装,阐述全新建筑设计生成方法。

一个包含Euler函数及k阶Smarandache ceil函数的方程及其正整数解

设k≥2为给定的整数.对任意正整数n,k阶Smarandache ceil函数Sk(n)定义为Sk(n)=min{x:x∈N,n|xk}.本文的主要目的是利用初等方法研究函数方程Sk(n)=φ(n)的可解性,并给出该方程的所有正整数解,其中φ(n)为Euler函数.

关于Smarandache Ceil函数的一个混合均值

对任意正整数n,k≥2为给定整数,Smarandache Ceil函数sk(n)定义为最小的正整数x,使得n|xk,即Sk(n)=min{x∶x∈N,n|xk}.利用Smarandache Ceil函数的定义及解析的方法,研究Smarandache Cei函数sk(n)与欧拉函数的均值分布性质,并给出一个有趣的渐近公式.