Occlusive cerebrovascular disease leads to brain ischemia that causes neurological deficits. Here we introduce a new strategy combining mesenchymal stromal cells (MSCs) and ex vivo hepatocyte growth factor (HGF) gene transferring with a multimutated herpes simplex virus type-1 vector in a rat transient middle cerebral artery occlusion (MCAO) model. Gene-transferred MSCs were intracerebrally transplanted into the rats' ischemic brains at 2 h (superacute) or 24 h (acute) after MCAO. Behavioral tests showed significant improvement of neurological deficits in the HGF-transferred MSCs (MSC-HGF)-treated group compared with the phosphatebuffered saline (PBS)-treated and MSCs-only-treated group. The significant difference of infarction areas on day 3 was detected only between the MSC-HGF group and the PBS group with the superacute treatment, but was detected among each group on day 14 with both transplantations. After the superacute transplantation, we detected abundant expression of HGF protein in the ischemic brain of the MSC-HGF group compared with others on day 1 after treatment, and it was maintained for at least 2 weeks. Furthermore, we determined that the increased expression of HGF was derived from the transferred HGF gene in gene-modified MSCs. The percentage of apoptosis-positive cells in the ischemic boundary zone (IBZ) was significantly decreased, while that of remaining neurons in the cortex of the IBZ was significantly increased in the MSC-HGF group compared with others. The present study shows that combined therapy is more therapeutically efficient than MSC cell therapy alone, and it may extend the therapeutic time window from superacute to acute phase.

Heat shock transcription factor 1 (HSF1) monitors the structural integrity of intracellular proteins and its regulation is essential for the health and longevity of eukaryotic organisms. HSF1 also plays a role in the acute inflammatory response in the negative regulation of cytokine gene transcription. Here we show, for the first time, that HSF1 is regulated by the proinflammatory protein kinase MAPKAP kinase 2 (MK2). We have shown that MK2 directly phosphorylates HSF1 and inhibits activity by decreasing its ability to bind the heat shock elements (HSE) found in the promoters of target genes encoding the HSP molecular chaperones and cytokine genes. We show that activation of HSF1 to bind HSE in hsp promoters is inhibited through the phosphorylation of a specific residue, serine 121 by MK2. A potential mechanism for MK2-induced HSF1 inactivation is suggested by the findings that phosphorylation of serine 121 enhances HSF1 binding to HSP90, a major repressor of HSF1. Dephosphorylation of serine 121 in cells exposed to non-steroidal anti-inflammatory drugs leads to HSP90 dissociation from HSF1, which then forms active DNA binding trimers. These experiments indicate a novel mechanism for the regulation of HSF1 by proinflammatory signaling and may permit HSF1 to respond rapidly to extracellular events, permitting optimal physiological regulation.Heat shock factor 1 (HSF1) 3 is the transcriptional activator of HSP molecular chaperone genes during stress (1, 2). The hsf1 gene plays an essential role in protection of cells from heat shock by regulating the induction of cytoprotective HSP and in protection against the effects of endotoxins through its ability to repress the transcription of proinflammatory cytokines through inhibition of factors involved in cytokine transcription such as 4). Aging is associated with the degeneration of HSP expression with time and the loss of resistance to cellular oxidants; elevated HSF1 leads to significant increase in lifespan in Caenorhabditis elegans (5-7). In cancer, the converse situation applies, and malignant transformation is associated with aberrantly high levels of HSP (8, 9). These clinical phenomena reflect the role of HSP molecular chaperones in cellular regulation, as either up-or downregulation of HSP expression can profoundly modulate multiple key proteins within the cell (10). It is therefore clear that elucidating the molecular mechanisms that control HSP expression in mammalian cells is essential.Under normal conditions, most HSF1 is inactive and maintained in a compacted monomeric form (11-13). Inactive HSF1 lacks the ability to bind to the heat shock elements (HSE) in hsp promoters, is unable to trans-activate hsp genes and fails to repress the promoters of proinflammatory cytokines (13-16). Activation of HSF1 is a complex process involving monomer to trimer transition and DNA binding; hyperphosphorylation, and capacity to activate target promoters (12,(17)(18)(19). Trimerization of HSF1 is governed by leucine zipper domains in the amino terminus and is subject to ...

Tanshinone IIA is a fat-soluble pharmacologically active ingredient of Danshen, a well-known traditional Chinese medicine used for cardiovascular diseases such as coronary heart disease. Tanshinone IIA has been confirmed to suppress miR-1 and reduce the arrhythmogenesis after myocardial infarction (MI). However, the modulation mechanism is not clear. Tanshinone IIA was administrated daily for 7 days before ligation of the left anterior descending artery (LAD) and lasted for 3 months after LAD. Neonatal cardiomyocytes were exposed to 2% O₂+95% N₂ condition for 24 h to simulate ischemia in vivo. Protein expression was examined with Western blot and miR-1 level was quantified by Real-time PCR. Our results showed that tanshinone IIA relieved ischemia-induced injury by improving the cardiac function. This beneficial effect may due to the depression of the elevated miR-1 level in ischemic and hypoxic cardiomyocytes, which subsequently restored its target Cx43 protein. Furthermore, tanshinone IIA could inhibit activated p38 MAPK and heart special transcription factors SRF and MEF2, in ischemic and hypoxic cardiomyocytes. Pretreatment with p38 MAPK inhibitor, SB203580 (10 uM), significantly relieved hypoxia-induced miR-1 increment and restored its downstream target Cx43 protein expression. These data suggest that tanshinone IIA play a role in protection cardiomyocytes from ischemic and hypoxic injury. The effect is based on inhibiting miR-1 expression through p38 MAPK signal pathway. This might provide us a new target to explore the novel strategy for ischemic cardioprotection.

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Background and Purpose-Fibroblast growth factor-2 (FGF-2) administration and bone marrow stromal cell (MSC) transplantation could improve neurological deficits after occlusive cerebrovascular disease. In the present study, we examined the effects of neurological improvement after transient middle cerebral artery occlusion (MCAO) in rats by a novel therapeutic strategy with FGF-2 gene-transferred MSCs by the herpes simplex virus type 1 (HSV-1) vector. Methods-Adult Wistar rats were anesthetized. Nonmodified MSCs, FGF-2-modified MSCs with HSV-1 1764/-4/pR19/ ssIL2-FGF-2, or PBS was administered intracerebrally 24 hours after transient right MCAO. All animals underwent behavioral tests for 21 days, and the infarction volume with 2-3-5-triphenylterazolium was detected 3 days and 14 days after the MCAO. Three days and 7 days after the MCAO, the FGF-2 production in the ipsilateral hemisphere of the MCAO was measured with ELISA. Seven and 14 days after the MCAO, immunohistochemical staining for FGF-2 was applied. Results-The stroke animals receiving FGF-2-modified MSCs demonstrated significant functional recovery comparedwith the other groups. Fourteen days after the MCAO, there was a significant reduction in infarction volume only in FGF-2-modified MSC-treated group. FGF-2 production in the FGF-2-modified MSC-treated brain was significantly higher compared with the other groups at 3 and 7 days after MCAO. Administrated FGF-2-modified MSCs strongly expressed the FGF-2 protein, which was proven by ELISA. Conclusions-Our

Genetic Algorithm (GA) is globally oriented in searching and thus useful in optimizing multiobjective problems, especially where the objective functions are ill-defined. Conceptual rainfall-runoff models that aim at predicting streamflow from the knowledge of precipitation over a catchment have become a basic tool for flood forecasting. The parameter calibration of a conceptual model usually involves the multiple criteria for judging the performances of observed data. However, it is often difficult to derive all objective functions for the parameter calibration problem of a conceptual model. Thus, a new method to the multiple criteria parameter calibration problem, which combines GA with TOPSIS (technique for order performance by similarity to ideal solution) for Xinanjiang model, is presented. This study is an immediate further development of authors' previous research (Cheng, C.T., Ou, C.P., Chau, K.W., 2002. Combining a fuzzy optimal model with a genetic algorithm to solve multi-objective rainfall-runoff model calibration. Journal of Hydrology, 268, 72-86), whose obvious disadvantages are to split the whole procedure into two parts and to become difficult to integrally grasp the best behaviors of model during the calibration procedure. The current method integrates the two parts of Xinanjiang rainfall-runoff model calibration together, simplifying the procedures of model calibration and validation and easily demonstrated the intrinsic phenomenon of observed data in integrity. Comparison of results with two-step procedure shows that the current methodology gives similar results to the previous method, is also feasible and robust, but simpler and easier to apply in practice. q

Recent research found that Tiron was an effective antioxidant that could act as the intracellular reactive oxygen species (ROS) scavenger or alleviate the acute toxic metal overload in vivo. In this study, we investigated the inhibitory effect of Tiron on matrix metalloproteinase (MMP)-1 and MMP-3 expression in human dermal fibroblast cells. Western blot and ELISA analysis revealed that Tiron inhibited ultraviolet B (UVB)-induced protein expression of MMP-1 and MMP-3. Real-time quantitative PCR confirmed that Tiron could inhibit UVB-induced mRNA expression of MMP-1 and MMP-3. Furthermore, Tiron significantly blocked UVB-induced activation of the MAPK signaling pathway and activator protein (AP)-1 in the downstream of this transduction pathway in fibroblasts. Through the AP-1 binding site mutation, it was found that Tiron could inhibit AP-1-induced upregulation of MMP-1 and MMP-3 expression through blocking AP-1 binding to the AP-1 binding sites in the MMP-1 and MMP-3 promoter region. In conclusion, Tiron may be a novel antioxidant for preventing and treating skin photoaging UV-induced.

The importance of the environmental impact assessment (EIA) of large development projects is increasingly underlined. Usually, EIA involves a lot of qualitative and quantitative criteria. Data Envelopment Analysis (DEA), an effective method which is used to rank and select the best alternative from a set of alternatives, is not tailored to address qualitative criteria, thus rendering the application to multiple criteria problems not amenable. This paper presents a new methodology of Multiple Criteria Data Envelopment Analysis (MCDEA) which can address both qualitative and quantitative criteria. MCDEA is divided into two-stage for fully ranking units and each unit has multiple inputs and outputs. In the first stage, a qualitative method is applied to compare the qualitative performance of alternatives. Then MCDEA is used to rank the alternatives by considering the relative membership degree of qualitative factors as one of the quantitative data. A case study on the selection of dam location illustrates the effectiveness of the proposed methodology.