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To investigate the nutrient limiting factors for a practical fertilization proposal on direct - sowing oilseed rape in Guizhou Province, fertilization field experiments were conducted using cultivar Youyan 599 (Brassica napus L.) in main rapeseed planting area of Guizhou during 2011 - 2012. Results showed that compared to no fertilization treatment (CK), seed yield of direct - sowing oilseed rape significantly increased when fertilizers were applied. Among them, NPKB treatment had the best performance with the average yield of 2189kg/hm². While compared to NPKB treatment, yield outputs decreased to 961, 342, 295 and 184kg/hm² in PKB (without N), NKB (without P), NPB (without K), and NPK (without B) treatments, respectively. It indicated that the nutrient limiting factors were ranked as N > P > K > B. Compared to farmers' fertilization practices (FFP ) treatment, NPKB treatment increased the application rate, but the production was improved significantly by 430kg/hm². Nutrient accumulation and fertilizer use efficiency were also significantly improved. Nitrogen, phosphorus and potassium nutrient accumulation (corresponding fertilizer use efficiency) increased 28.6 kg/hm² (13.5%), 9.6 kg/hm² (3.3%), 71.4 kg/hm² (23.9%) compared to FFP . As for economic profits, the revenue of NPKB treatment was increased by 1 109 yuan/hm2 compared to FFP. It suggested that the yield, nutrients accumulation and profits could be significantly improved by a combination of balanced fertilization. [ABSTRACT FROM AUTHOR]

Lipid nanoparticles with solid matrix have been given increasing attention due to their biodegradable status and ability to entrap a variety of biologically active compounds. In this study, new phospholipid-based gelatin nanoparticles encapsulating basic fibroblast growth factor (bFGF) were developed to target the brain via nasal administration. Treatment effects were assessed by quantifying rotational behavior, monoamine neurotransmitter levels and tyrosine hydroxylase expression in 6-hydroxydopamine induced hemiparkinsonian rats. The gelatin nanostructured lipid carriers (GNLs) were prepared by a water-in-water emulsion method and then freeze-dried. The GNLs possessed better profile than gelatin nanoparticles (GNs), with particle size 143±1.14nm and Zeta potential −38.2±1.2mV. The intranasal GNLs efficiently enriched exogenous bFGF in olfactory bulb and striatum without adverse impact on the integrity of nasal mucosa and showed obvious therapeutic effects on hemiparkinsonian rats. Thus, GNLs are attractive carriers for nose-to-brain drug delivery, especially for unstable macromolecular drugs such as bFGF. From the Clinical Editor: This team of authors reports the development of phospholipid-based gelatin nanoparticles encapsulating basic fibroblast growth factor to target the brain via intranasal administration. A rat model of hemiparkinsonism was applied demonstrating a good safety profile and an obvious therapeutic effect. [ABSTRACT FROM AUTHOR]

A field trial was conducted in Dangyang City to compare the yield and nitrogen (N) use efficiency of 2 japonica rice varieties ( Zhongdao 1 and Huaidao 5) in the hilly land of central Hubei Province ( single-cropping indica rice area) under different N levels. The results showed that the grain yield of the treatments with N were increased averagely by 2 179 kg/hm² and 3 209 kg/hm², and the increase rates were 45.6% and 53.5%, respectively. Linear and platform correlations had been shown between yield and N fertilizer amount. The platform yields of "Zhongdao 1" and "Huaidao 5" were 7 517 kg/hm² and 10 234 kg/hm², respectively. The most suitable N application amount were 221.0 kg/hm² and 235.5 kg/hm², respectively. Compared with N0 treatment, the N uptakes by these 2 japonica rice varieties were averagely increased 58.7 kg/hm² and 71.8 kg/hm², and the increase rates were 80.9% and 85.2%. When the N application amount was 248 kg/hm², the N recovery efficiency (NRE), N agronomic efficiency (NAE), N contribution rate (NCR) and partial factor productivity of applied N (PFPN) of the 2 varieties were 29.4% and 37.6%, 10.6 kg/kg and 18.0 kg/kg, 35.4% and 42.6%, 29.8 kg/kg and 42.3 kg/kg. The result of comprehensive analysis believed that " Huaidao 5" could get a higher yield and high N use efficiency when planted in the hilly land of central Hubei Province. [ABSTRACT FROM AUTHOR]

Using curcumol that was extracted from the volatile oil of Rhizoma Curcumae as the raw material, its derivatives were synthesized and purified. The structures of these compounds were confirmed by1H,13C NMR, and mass spectral data. The test compounds were evaluated for theirin vitroanti-tumor activity against gastric cancer cell lines SGC-7901 and lung carcinoma cell line H460 by methyl thiazolyl tetrazolium chromatometry. Distinct structure–activity relationships of these curcumol derivatives were also revealed for inhibiting cell proliferation. Presence of electron-withdrawing groups or amino could increase the activity significantly, whereas esterification of 8-hydroxy diminished the anti-tumor activity. Many of the tested candidates exhibited higher inhibition efficiency than curcumol, suggesting that structural modifications could enhance its activity effectively. [ABSTRACT FROM AUTHOR]

The role of autophagy in the recovery of spinal cord injury remains controversial; in particular, the mechanism of autophagy regulated degradation of ubiquitinated proteins has not been discussed to date. In this study, we investigated the protective role of basic fibroblast growth factor (bFGF) both in vivo and in vitro and demonstrated that excessive autophagy and ubiquitinated protein accumulation is involved in the rat model of trauma. bFGF administration improved recovery and increased the survival of neurons in spinal cord lesions in the rat model. The protective effect of bFGF is related to the inhibition of autophagic protein LC3II levels; bFGF treatment also enhances clearance of ubiquitinated proteins by p62, which also increases the survival of neuronal PC-12 cells. The activation of the downstream signals of the PI3K/Akt/mTOR pathway by bFGF treatment was detected both in vivo and in vitro. Combination therapy including the autophagy activator rapamycin partially abolished the protective effect of bFGF. The present study illustrates that the role of bFGF in SCI recovery is related to the inhibition of excessive autophagy and enhancement of ubiquitinated protein clearance via the activation of PI3K/Akt/mTOR signaling. Overall, our study suggests a new trend for bFGF drug development for central nervous system injuries and sheds light on protein signaling involved in bFGF action. [ABSTRACT FROM AUTHOR]

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Hypertrophic scars (HTS) and keloids are challenging problems. Their pathogenesis results from an overproduction of fibroblasts and excessive deposition of collagen. Studies suggest a possible anti-scarring effect of basic fibroblast growth factor (bFGF) during wound healing, but the precise mechanisms of bFGF are still unclear. In view of this, we investigated the therapeutic effects of bFGF on HTS animal model as well as human scar fibroblasts (HSF) model. We show that bFGF promoted wound healing and reduced the area of flattened non-pathological scars in rat skin wounds and HTS in the rabbit ear. We provide evidence of a new therapeutic strategy: bFGF administration for the treatment of HTS. The scar elevation index (SEI) and epidermal thickness index (ETI) was also significantly reduced. Histological reveal that bFGF exhibited significant amelioration of the collagen tissue. bFGF regulated extracellular matrix (ECM) synthesis and degradation via interference in the collagen distribution, the α-smooth muscle actin (α-SMA) and transforming growth factor-1 (TGF-β1) expression. In addition, bFGF reduced scarring and promoted wound healing by inhibiting TGFβ1/SMAD-dependent pathway. The levels of fibronectin (FN), tissue inhibitor of metalloproteinase-1 (TIMP-1) collagen I, and collagen III were evidently decreased, and matrix metalloproteinase-1 (MMP-1) and apoptosis cells were markedly increased. These results suggest that bFGF possesses favorable therapeutic effects on hypertrophic scars in vitro and in vivo, which may be an effective cure for human hypertrophic scars. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the progressive loss of the dopaminergic neurons; however, its crucial mechanism of the metabolic changes of neurotransmitters remains ambiguous. The pathological mechanism of PD might involve cerebral metabolism perturbations. In this study, ex vivo proton nuclear magnetic resonance (H NMR) was used to determine the level changes of 13 metabolites in the bilateral striatum of 6-hydroxydopamine (6-OHDA)-induced PD rats. The results showed that, in the right striatum of 6-OHDA-induced PD rats, increased levels of glutamate (Glu) and γ-aminobutyric acid (GABA) concomitantly with decreased level of glutamine (Gln) were observed compared to the control. Whereas, in the left striatum of 6-OHDA-induced PD rats, increased level of Glu with decreased level of GABA and unchanged Gln were observed. Other cerebral metabolites including lactate, alanine, creatine, succinate, taurine, and glycine were also found to have some perturbations. The observed metabolic changes for Glu, Gln, and GABA are mostly likely the result of a shift in the steady-state equilibrium of the Gln-Glu-GABA metabolic cycle between astrocytes and neurons. The altered Gln and GABA levels are most likely as a strategy to protect neurons from Glu excitotoxic injury after striatal dopamine depletion. Changes in energy metabolism and tricarboxylic acid cycle might be involved in the pathogenesis of PD. [ABSTRACT FROM AUTHOR]

Aim To investigate the mechanism of endoplasmic reticulum (ER) stress-induced apoptosis as well as the protective action of basic fibroblast growth factor ( bFGF) both in vivo and in vitro. Methods and Results ER stress-induced apoptosis was involved in the injuries of spinal cord injury (SCI) model rat. bFGF administration improved the recovery and increased the survival of neurons in spinal cord lesions in model rat. The protective effect of bFGF is related to the inhibition of CHOP, GRP78 and caspase-12, which are ER stress-induced apoptosis response proteins. bFGF administration also increased the survival of neurons and the expression of growth-associated protein 43 (GAP43), which is related to neural regeneration. The protective effect of bFGF is related to the activation of downstream signals, PI3K/Akt/GSK-3β and ERK1/2, especially in the ER stress cell model. Conclusions This is the first study to illustrate that the role of bFGF in SCI recovery is related to the inhibition of ER stress-induced cell death via the activation of downstream signals. Our work also suggested a new trend for bFGF drug development in central neural system injuries, which are involved in chronic ER stress-induced apoptosis. [ABSTRACT FROM AUTHOR]

Experiments in vitro and in vivo were designed to investigate tumor growth inhibition of chemotherapeutics-loaded liposomes enhanced by acoustic cavitation. Doxorubicin-loaded liposomes (DOX liposomes) were used in experiments to investigate acoustic cavitation mediated effects on cell viability and chemotherapeutic function. The influence of lingering sensitive period after acoustic cavitation on tumor inhibition was also investigated. Animal experiment was carried out to verify the practicability of this technique in vivo. From experiment results, blank phospholipid-based microbubbles (PBM) combined with ultrasound (US) at intensity below 0.3 W/cm2 could produce acoustic cavitation which maintained cell viability at high level. Compared with DOX solution, DOX liposomes combined with acoustic cavitation exerted effective tumor inhibition in vitro and in vivo. The lingering sensitive period after acoustic cavitation could also enhance the susceptibility of tumor to chemotherapeutic drugs. DOX liposomes could also exert certain tumor inhibition under preliminary acoustic cavitation. Acoustic cavitation could enhance the absorption efficiency of DOX liposomes, which could be used to reduce DOX adverse effect on normal organs in clinical chemotherapy. [ABSTRACT FROM AUTHOR]

Polymeric nanoparticles are widely used as targeted carriers for biomacromolecules. In this paper, modified gelatin nanoparticles were prepared and their feasibility as insulin pulmonary administration system was investigated. d, l-glyceraldehyde and poloxamer 188 were used for gelatin nanoparticle preparation. Novel water-in-water emulsion technique was used to prepare insulin-loaded nanoparticles. Morphological examination of insulin-loaded nanoparticles was carried out using scanning electron microscopy (SEM). Intratracheal instillation of insulin-loaded nanoparticles was performed to evaluate animal hypoglycemic effect. With fluorescence labeling of insulin, alveolar deposition and absorption of insulin-loaded nanoparticles were investigated. Histological changes in the lung were also observed to evaluate the safety. From the micromorphology observation, insulin-loaded nanoparticles under gelatin-poloxamer 188 ratio at 1:1 showed smooth and uniform surface, with average particle size 250 nm and Zeta potential −21.1 mV. From animal experiment, insulin-loaded nanoparticles under gelatin-poloxamer 188 ratio at 1:1 promoted insulin pulmonary absorption effectively and showed good relative pharmacological bioavailability. Proved by alveolar deposition result, FITC-insulin-loaded nanoparticle group was characterized by an acute and rapid hypoglycemic effect. In addition, nanoparticles could guarantee the safety of lung by reducing insulin deposition in lung. A transient weak inflammatory response was observed at 1 day after administration. With good physical characterization, high bioavailability, fast and stable hypoglycemic effect, insulin-loaded nanoparticles might be developed as a novel insulin pulmonary system for diabetes therapy. [ABSTRACT FROM AUTHOR]

Abstract: After artificial hatching Locusta migratoria manilensis Meyen, the male adults were cultured in different temperature. By flow cytometry, DNA contents of apoptotic cells of germ cells were tested and analysed. The results showed that the apoptotic cells under different temperature always appeared in spermatogenesis of Locusta migratoria manilensis Meyen but arrived the peak in 45°C. The conclusion of the study showed that the 25°C was the best temperature of development and the 45°C was the worst temperature of development in spermatogenesis of Locusta migratoria manilensis Meyen. [Copyright &y& Elsevier]

In order to facilitate the intracellular delivery of therapeutic agents, a new type of liposomes-propylene glycol liposomes (PGL) were prepared, and their cell translocation capability in vitro was examined. PGL was composed of hydrogenated egg yolk lecithin, cholesterol, Tween 80 and propylene glycol. With curcumin as a model drug, characterization of loaded PGL were measured including surface morphology, particle size, elasticity, encapsulation efficiency of curcumin and physical stability. Using curcumin-loaded conventional liposomes as the control, the cell uptake capacity of loaded PGL was evaluated by detection the concentration of curcumin in cytoplasm. Compared with conventional liposomes, PGL exhibited such advantages as high encapsulation efficiency (92.74% ± 3.44%), small particle size (182.4 ± 89.2 nm), high deformability (Elasticity index == 48.6) and high stability both at normal temperature (about 25°C) and low temperature at 4°C. From cell experiment in vitro, PGL exhibited the highest uptake of curcumin compared with that of conventional liposomes and free curcumin solution. Little toxic effect on cellular viability was observed by methyl tetrazolium assay. In conclusion, PGL might be developed as a promising intracellular delivery carrier for therapeutic agents. [ABSTRACT FROM AUTHOR]

The article presents a study which examines a method of high-density fermentation of rhCu, Zn-SOD Pichia pastoris.

Field trials in two sites were conducted to study the effect of special fertilizer for rapeseed on the growth, yield, nutrient uptake and nutrient recovery efficiency. Results showed that application of multi - nutrient and long - effect special fertilizer for rapeseed promoted growth significantly. Plant height, stem diameter, SPAD value, branch number and dry weight of individual plant increased by 30. 1%, 72.7%, 162.2%, 31.0% and 130. 1% respectively, compared to that of no fertilizer control. Significant linear and platform correlations had been shown between yield and fertilizer quantity. Proper fertilizer increased yield significantly, for it promoted branch number and - number. Results also showed that dry matter and N, P2O5 and K2O uptake by rapeseed were all improved significantly after using the special fertilizer, although the nutrient recovery efficiency decreased gradually when fertilizer quantity increased. The average N, P2O5 and K2O recovery efficiency on two experimental sites were 37.0%, 33.5% and 75.6% respectively when fertilizer quantity was 1 125kg/hm2. It could be calculated that the appropriate quantity of multi - nutrient and long - effect special fertilizer for rapeseed was 1 200 to 1 275 kg/hm2 to target yield of 2 700 to 3 000kg/hm2. [ABSTRACT FROM AUTHOR]

Field experiments were carried out to study the effects of nitrogen ( N ) application rate on rapeseed yield and N fertilizer use efficiency in Hubei Province during 2007 to 2008. Compared to no N application, the yield of N treatments ( as 75, 150 and 225kg/hm²) increased 41.9%, 70.3% and 66.2% respectively. N content increased 9.1%, 14.2% and 13.1%, and total N absorbed by above ground parts increased 59.6%, 111.6% and 108.0% respectively. The results demonstrated that N application significant promoted growth, yield and N accumulation. With increased N application rate, N requirement for 100kg -seeds was increased, while N agronomic use efficiency (AUE), partial factor productivity (PFP) and apparent recovery efficiency (ARE) of rapeseed decreased significantly. Based on rapeseed yield and N use efficiency, the optimum N fertilizer application rate was 150kg/hm² in Hubei Province. [ABSTRACT FROM AUTHOR]

In order to evaluate the expression of oleosin - bFGF fusion protein in plant oil body, polymerase chain reaction amplification was used to clone a soybean oleosin promoter Ddprm, a soybean oleosin gene (Ddoil) and a bFGF gene which was changed with plant codon usage. All these were used to construct a plant binary expression vector p1390 Ddprm - Ddoil - bFGF, and Agrobacterium - mediated transformation of Arabidopsis thaliana was conducted by floral dip. The results showed that bFGF gene integrated into Arabidopsis genome. The total protein of the T2 seeds were extracted for SDS - PAGE, Western blot, ELISA and cell proliferation activity to further assess the oleosin -bFGF fusion protein. [ABSTRACT FROM AUTHOR]

In order to recommend fertilizer application for different rapeseed, the most popular 34 cultivars from Yangtze River area were planted in field at Shayang, Hubei province, with the same fertilization level (N210kg/hm2P2O5 45kg/hm2K2O 67. 5kg/hm2). The materials were divided into 4 types according to their nitrogen uptake and efficiency. Type A cultivars had both high uptake and utilization efficiency, type B had low uptake but high utilization efficiency, type C had high uptake but low utilization efficiency, and type D had both low uptake and utilization efficiency. Their agronomic traits, dry matter accumulation and N accumulation were investigated. Results showed that type A had strong advantage on inflorescence length, pods on main inflorescence, first branch number and pods of first branches. The higher the nitrogen uptake efficiency, the more dry matter and nitrogen accumulated. The accumulated quantity of dry mass and nitrogen was shown as C (and A) > D (and B). More dry matter and nitrogen were distributed into seeds and less into seed coats in high nitrogen utilization efficiency type A and B. The seed yield and seed nitrogen accumulation were shown as A > B (and C) > D. [ABSTRACT FROM AUTHOR]

Objective: An in vivo study on enhancing antitumor effect of intravenous epirubicin hydrochloride (EPI) by acoustic cavitation in situ combined with phospholipid-based microbubbles (PMB) was reported. Methods: Five-week-old male nude mice were used, and HCT-116 cells were s.c. (subcutaneous injection) inoculated in axilla of these mice. Five groups were designed and five consecutive treatments were applied to investigate the tumor growth, body weight growth, and normal organ growth. Results: Inhibition effects on tumor growth were observed in all groups treated by EPI. However, the potency of tumor growth retardation caused by EPI depended on the application of PMB and US in situ. An effective retardation on tumor growth and improved mice survival status were observed when intravenous EPI combined with sonicated PMB in situ. Short-term lingering sensitive period for chemotherapeutic drugs after acoustic cavitation was also observed in experiment under asynchronous administration of EPI and sonicated PMB. Conclusion: Intravenous EPI combined with PMB-mediated cavitation in situ might provide a potential application for US-mediated chemotherapy. [ABSTRACT FROM AUTHOR]

Field experiments were carried out to study the effects of phosphorus (P) application rate on rapeseed yield and P fertilizer use efficiency in Hubei Province during 2007 -2008. Compared to the control with zero P application, the yield of P2O5 treatments (as 37. 5, 75. 0 and 112. 5kg/hm2, respectively) increased 24. 2% 42.9% and 43. 2% ,as well as the P content increased 10. 5%, 16. 7% and 27.1%, and total P absorbed by aboveground parts increased 36. 1% , 59.1% and 78.7% , respectively. The results indicated significant promotion of P application to growth, yield and P accumulation. With the increase of P application rate, the P required for 100kg seeds was increased, while the P agronomic use efficiency (AUE) , partial factor productivity (PFP) , apparent recovery efficiency (ARE) and physiological use efficiency (PUE) of rapeseed were decreased significantly. 75.0kg/hm2 of P fertilizer was the optimum to both yield and P use efficiency. [ABSTRACT FROM AUTHOR]

For the sake of understanding the deterioration behavior of concrete in actual railway tunnel structures subjected to aggressive sulfate medium in practice, detailed field investigations and tested analysis on sprayed concrete linings of approximately 40-year-old railway tunnels in environments containing sulfate ion were carried out, respectively. The results show that the deterioration of concretes in the investigated area is serious, which involves complicated physicochemical process between the sulfate salt and concrete. Among them, the secondary sulfate minerals such as gypsum formation under very high concentration sulfate ion condition by accumulating and evaporation process dominate, followed by the crystallization of sulfate salt and formation of thaumasite. [ABSTRACT FROM AUTHOR]

An in vivo study on enhancing epirubicin hydrochloride (EPI) inhibition on tumor growth by ultrasound (US) was reported. Five-week-old male nude mice were used and HL-60 cells were s.c. (subcutaneous injection) inoculated in axilla of these mice. Six groups were designed and five consecutive treatments were applied to investigate the inhibition on tumor growth and body weight growth. US applied locally to the tumor resulted in a substantially increased drug uptake in tumor cells. The inhibition on tumor growth depended on the position of drug injection and phospholipid-based microbubble (PMB) application. Tumor growth rate under group 1 (PMB+US) was similar to that of blank control. The order of the inhibition on tumor volume growth was: group 4 (s.c. EPI+PMB+US) > group 5 intraperitoneal (i.p. EPI+PMB+US) > group 2 (i.p. EPI) > group 3 (s.c. EPI+US) > group 1 (PMB+US). Similar conclusion was obtained from experimental measurements of tumor weight change. The order of animal survival status for EPI administration groups was: group 4 > group 5 > group 2 > group 3. Chemotherapeutic drug inhibition on tumor growth could be enhanced by local US combined with PMB, which might provide a potential application for US-mediated chemotherapy. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the factors for enhancing the susceptibility of cancer cells to chemotherapeutic drug by ultrasound microbubbles. Ultrasound (US) combined with phospholipid-based microbubbles (MB) was used to enhance the susceptibility of colon cancer cell line SWD-620 to anticancer drugs Topotecan hydrochloride (TOP). Experiments were designed to investigate the influence of main factors on cell viability and cell inhibition, such as US intensity, MB concentration, drug combination with MB, asynchronous action between US triggered cavitation and drug entering cell, MB particle size. US exposure for 10 sec with US probe power at 0.6 W/cm2 had satisfied cell viability. Treated with US combined with 15% MB, cell viability maintained more than 85% and cell inhibition 86.16%. Under optimal US combined with MB, TOP showed much higher cell inhibition than that of only TOP group. Cell inhibition under short delayed time (<2 h) for TOP addition did not show obvious difference. In terms of MB particle size, the order of cell inhibition was: Mixture > Micron bubble part > Nanometer bubble part. US combined with MB can enhance the susceptibility of cancer cells to chemotherapeutic drug, which may provide a potential method for US-mediated tumor chemotherapy. [ABSTRACT FROM AUTHOR]

Both fibroblast growth factor 2 (FGF2) and luteinizing hormone (LH) have been reported to regulate androgen production in Leydig cells in progenitor Leydig cells. The objective of the present study is to examine the regulation of androgen production in rat immature Leydig cells (ILCs). ILCs were isolated from 35-day-old rat testes and cultured in DMEM/F12 medium with LH (1 ng/ml) or FGF2 (10 ng/ml). 5α-Androstane-3α, 17β-diol (3α-DIOL), the primary androgen in ILCs, and testosterone (T) were measured by Radioimmuno assay. The results showed the LH stimulated androgen production in ILCs, and FGF2 did not. However, FGF2 decreased the LH-stimulated androgen production. Real-time PCR and enzyme assay showed that FGF2 decreased levels of several steroidogenic enzymes, inhibited the expressions of steroidogenic acute regulatory (StAR) protein and steroidogenic factor 1 ( Nr5a1) in LH-stimulated ILCs. FGF2-mediated inhibition of Nr5a1gene expression may be the mechanism through which FGF2 inhibits LH-stimulated androgen production. [ABSTRACT FROM AUTHOR]

Abstract: Recently our laboratory reported evidence showing that hNUDC acts as an additional cytokine for thrombopoietin receptor (Mpl). Previously known as the human homolog of a fungal nuclear migration protein, hNUDC plays a critical role in megakaryocyte differentiation and maturation. Here we sought to further clarify the hNUDC-Mpl ligand–receptor relationship by utilizing interference RNA (RNAi) to knockdown Mpl expression in a megakaryocyte cell line. We created U6 promoter driven constructs to express short hairpin RNAs (shRNA) with affinity for different sites on Mpl mRNA. By including Mpl-EGFP fusion protein in these constructs, we were able to effectively screen the shRNA that was most efficient in inhibiting Mpl mRNA expression. This shRNA was subsequently transferred into a lentivirus vector and transduced into Dami cells, a cell line which constitutively expresses endogenous Mpl. This lentiviral vector was also designed to simultaneously express EGFP to monitor transfection efficiency. Our results show that lentivirus can be used to effectively deliver shRNAs into Dami cells and cause specific inhibition of Mpl protein expression after transduction. Furthermore, we show the functional effects of shRNA-mediated Mpl silencing by demonstrating reduced hNUDC stimulated megakaryocyte proliferation and differentiation. Thus, the use of a RNAi knockdown strategy has allowed us to pinpoint the connection of hNUDC with Mpl in the regulation of megakaryocyte maturation. [Copyright &y& Elsevier]

Abstract: Objective: To investigate the protective effect of a mutant of acidic fibroblast growth factor (MaFGF) against cerebral ischaemia-reperfusion injury in rats. Methods: Sixty male Sprague–Dawley rats were randomly divided into six groups as follows: sham-operated group, untreated group, 20μg/kg, 40μg/kg and 80μg/kg MaFGF-treated groups and also the positive control group. Cerebral ischaemia-reperfusion injury was induced by middle cerebral artery occlusion (MCAO) for 2h followed by reperfusion for 24h. Different dose of MaFGF were infused intravenously at 1h after middle cerebral artery (MCA) occlusion. Nimodipine was used as positive control. The behaviour deficit score, brain-infarcted area, brain oedema degree, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were detected at 24h after reperfusion. Results: The results showed that MaFGF at the dose of 20μg/kg, 40μg/kg and 80μg/kg significantly alleviated brain injury. Compared to untreated group, the behaviour deficits were much less severe, the brain oedema alleviated obviously, the MDA contents decreased and SOD activity increased dramatically in MaFGF-treated groups respectively. The efficacy of MaFGF was similar to that of nimodipine. Conclusion: The results demonstrate that MaFGF has neuroprotective effect against brain injury resulting from focal ischaemia-reperfusion in Sprague–Dawley rats. [Copyright &y& Elsevier]

Objective: Glycogen synthase kinase (GSK)-3beta plays an important role in cardiomyopathies. Cardiac-specific metallothionein-overexpressing transgenic (MT-TG) mice were highly resistant to diabetes-induced cardiomyopathy. Therefore, we investigated whether metallothionein cardiac protection against diabetes is mediated by inactivation of GSK-3beta.Research Design and Methods: Diabetes was induced with streptozotocin in both MT-TG and wild-type mice. Changes of energy metabolism-related molecules, lipid accumulation, inflammation, nitrosative damage, and fibrotic remodeling were examined in the hearts of diabetic mice 2 weeks, 2 months, and 5 months after the onset of diabetes with Western blotting, RT-PCR, and immunohistochemical assays.Results: Activation (dephosphorylation) of GSK-3beta was evidenced in the hearts of wild-type diabetic mice but not MT-TG diabetic mice. Correspondingly, cardiac glycogen synthase phosphorylation, hexokinase II, PPARalpha, and PGC-1alpha expression, which mediate glucose and lipid metabolisms, were significantly changed along with cardiac lipid accumulation, inflammation (TNF-alpha, plasminogen activator inhibitor 1 [PAI-1], and intracellular adhesion molecule 1 [ICAM-1]), nitrosative damage (3-nitrotyrosin accumulation), and fibrosis in the wild-type diabetic mice. The above pathological changes were completely prevented either by cardiac metallothionein in the MT-TG diabetic mice or by inhibition of GSK-3beta activity in the wild-type diabetic mice with a GSK-3beta-specific inhibitor.Conclusions: These results suggest that activation of GSK-3beta plays a critical role in diabetes-related changes in cardiac energy metabolism, inflammation, nitrosative damage, and remodeling. Metallothionein inactivation of GSK-3beta plays a critical role in preventing diabetic cardiomyopathy. [ABSTRACT FROM AUTHOR]

Abstract: A convenient silver staining method for protein in sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) gels is described. The method is previsible, sensitive, and mass spectrometry (MS) compatible. Two visible counter ion dyes, ethyl violet (EV) and zincon (ZC), were used in the first staining solution with a detection limit of 2 to 8ng/band in approximately 1h. The dye-stained gel can be further stained by silver staining, which is based on acidic silver staining employing ZC with sodium thiosulfate as silver ion sensitizers. Especially, ZC has silver ion reducing power by cleavage of the diazo bond of the dye during silver reduction. The second silver staining can be completed in approximately 1h with a detection limit of 0.2ng/band. [Copyright &y& Elsevier]