hTERT-Driven Immortalization of RDEB Fibroblast and Keratinocyte Cell Lines Followed by Cre-Mediated Transgene Elimination

The recessive form of dystrophic epidermolysis bullosa (RDEB) is a crippling disease caused by impairments in the junctions of the dermis and the basement membrane of the epidermis. Using ectopic expression of hTERT/hTERT + BMI-1 in primary cells, we developed expansible cultures of RDEB fibroblasts and keratinocytes. We showed that they display the properties of their founders, including morphology, contraction ability and expression of the respective specific markers including reduced secretion of type VII collagen (C7). The immortalized keratinocytes retained normal stratification in 3D skin equivalents. The comparison of secreted protein patterns from immortalized RDEB and healthy keratinocytes revealed the differences in the contents of the extracellular matrix that were earlier observed specifically for RDEB. We demonstrated the possibility to reverse the genotype of immortalized cells to the state closer to the progenitors by the Cre-dependent hTERT switch off. Increased β-galactosidase activity and reduced proliferation of fibroblasts were shown after splitting out of transgenes. We anticipate our cell lines to be tractable models for studying RDEB from the level of single-cell changes to the evaluation of 3D skin equivalents. Our approach permits the creation of standardized and expandable models of RDEB that can be compared with the models based on primary cell cultures.     

A Modified Nonparametric Prewhitened Covariance Estimator

Abstract. This paper proposes a fully modified version of the spectral matrix estimator (and the long‐run variance estimator as a special case) proposed originally by Xiao and Linton [Journal of Time Series Analysis (2002) Vol. 23, pp. 215–250], and derives its asymptotic results. A striking feature of the modified spectral matrix estimator is to achieve the convergence rate of O(T ^?8/9) in the mean squared error (MSE), which is usually achieved under the fourth‐order spectral window. However, this estimator does not sacrifice the positive definiteness of the resulting estimate for the rate improvement; it is Hermitian and positive definite in finite samples by construction. The faster convergence rate is established by a multiplicative bias correction of the crude spectral estimator under the second‐order spectral window. The approximations to some sensible definitions of the MSE of the estimator and the bandwidths that minimize the asymptotic MSEs are also derived. Monte Carlo results indicate that for a wide variety of processes the modified spectral matrix estimator reduces the bias without inflating the variance and thus improves the MSE, compared with the crude, bias‐uncorrected estimator.     

电致孔经皮给药：表面活性剂对孔道存在时间和药物传输的影响

Electroporation creates aqueous pathways by short high-voltage pulses resulting in a transient perme- abilization of stratum corneum and an increase in the transdermal delivery rate.However the aqueous pathways will reseal after pulsing,which leads to the rapid drop of transdermal flux.In the present study,the surfactants were added to the donor solution to hinder the shrinkage and resealing of the electropore,and to prolong the lifetime of the aqueous pathways with the consideration that the surfactants could reduce the surface energy of the electropore. These effects of surfactants were demonstrated by the dynamic electrical resistance of the skin and the fluorescent imaging of the local transport regions.Piroxicam（PIX）was transported percutaneously in the presence of surfactants in vitro.Owing to the longer lifetime of aqueous pathways,together with the promotion of PIX availability at the barrier exterior and the improvement in the partition of PIX into the aqueous pathways,the presence of surfactants led to a remarkable increase in the transdermal delivery rate during electroporation and a significant growth of the accumulative transdermal amount of PIX.     

基于TW-SVM预测模型的某堆石坝变形预测分析

为提高监测资料有缺失的大坝变形预测模型精度,采用支持向量机方法建立一种具有小样本、高维、非线性的预测模型,并结合对其重要组成部分核函数的分析应用,提出一种根据结构风险最小化的TW-SVM预测模型。以某堆石坝为例进行研究,利用坝坡垂直位移和水平位移的监测数据,分别采用TW-SVM方法和BP神经网络(NET)方法建立相应预测模型进行比较分析。结果表明:采用TW-SVM方法和NET方法预测的垂直位移最大绝对误差分别为0.58 mm和6.18 mm,最大相对误差分别为270.00%和1 286.22%;采用TW-SVM方法和NET方法预测的水平位移最大绝对误差分别为0.25 mm和14.91 mm,最大相对误差分别为31.25%和1 189.85%;TW-SVM预测模型比NET预测模型更适合于影响因素为时间、水位的小样本预测分析。研究结果为堆石坝变形预测与分析提供参考。     

旁氏,夏天里的女人故事

美白产品始终是夏季护肤品市场竞争的焦点，旁氏推出“日夜美白不停顿”的新概念以应对传统美白产品的挑战，并利用策略发展和广告创意的源动力——“消费者洞察力”概念推出了一套精准的传播策略和创意出色的广告，对消费者产生了很强的吸引力。     

天然中草药美白祛斑化妆品

根据色斑的形成机理，采用相应的天然草本植物提取物淡化色斑。根据细胞内黑色素形成机理，选用适当抑制剂。减少黑色素的过量生成，防止皮肤表皮的继续黑化，提出美白与祛斑化妆品的合理配制方法。     

Nonparametric kernel density estimation near the boundary

Standard fixed symmetric kernel-type density estimators are known to encounter problems for positive random variables with a large probability mass close to zero. It is shown that, in such settings, alternatives of asymmetric gamma kernel estimators are superior, but also differ in asymptotic and finite sample performance conditionally on the shape of the density near zero and the exact form of the chosen kernel. Therefore, a refined version of the gamma kernel with an additional tuning parameter adjusted according to the shape of the density close to the boundary is suggested. A data-driven method for the appropriate choice of the modified gamma kernel estimator is also provided. An extensive simulation study compares the performance of this refined estimator to those of standard gamma kernel estimates and standard boundary corrected and adjusted fixed kernels. It is found that the finite sample performance of the proposed new estimator is superior in all settings. Two empirical applications based on high-frequency stock trading volumes and realized volatility forecasts demonstrate the usefulness of the proposed methodology in practice.     

个人洗护用品配方原理及应用技术(Ⅱ)--护肤品配方原理及应用

从护肤品配方原理、各类配方成分(如乳化剂)介绍和其应用等方面讨论护肤品开发应用的相关注意要点，并给出了参考配方。     

Hydrolysis of human horny cells by alkaline protease: Morphological observation of the process

The interaction of highly purified alkaline protease fromBacillus sp. KSM-K16 with the horny cells of human skin contained in skin grime was directly visualized by electron microscopy. It became clear that the protease first penetrates the horny cells and then adsorbs, mainly onto the internal structure of the cells at the initial stage of hydrolysis, and directly hydrolyzes the keratin filaments, though the marginal band surrounding them retains its original shape. Then, hydrolysate produced from the keratin filaments flows out of the cell, and early in the hydrolysis process keratin filaments decrease and then disappear, leaving a marginal band, i.e., the cell turns to a hollow state. As a result, the remaining marginal band loses support from inside the cell, thus promoting cleavage and dispersion. Until this stage in the protease reaction, the remarkable liberation of hydrolysis products as water-soluble protein does not occur.