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The uniformity of hot air flow inside the airflow dryer not only affects the moisture distribution at the outlet, but also affects the quality of the product. Based on the guide plate structure of the SH23A airflow tobacco dryer, a gradient curved guide plate dryer is designed, and the flow field distribution of the dryer is numerically investigated under different flow distribution conditions at the hot air inlet and flue gas inlet. The results show that the airflow uniformity is affected by the flow distribution at the inlet of the heated air and the inlet of the cigarette smoke, the structure of the guide plate, etc., the non-uniformity coefficient decreases with the increase of hot air inlet flow rate. The non-uniformity coefficient of tapered arc deflector decreases by 9–12 %.

Green manure application presents a valuable strategy for enhancing soil fertility and promoting ecological sustainability. By leveraging green manures for effective nitrogen management in agricultural fields can significantly reduce the dependency of primary crops on chemical nitrogen fertilizers, thereby fostering resource efficiency. This review examines the current advancements in the green manure industry, focusing on the modulation of nitrogen transformation in soil and how crops absorb and utilize nitrogen after green manure application. Initially, the influence of green manure on soil nitrogen transformation is delineated, covering processes such as soil nitrogen immobilization, and mineralization, and losses including NH3, N2O, and NO3−-N leaching. The review then delves into the effects of green manure on the composition and function of soil microbial communities, highlighting their role in nitrogen transformation. It emphasizes the available nitrogen content in the soil, this article discussing nitrogen uptake and utilization by plants, including aspects such as nitrogen translocation, distribution, the root system, and the rhizosphere environment of primary crops. This provides insights into the mechanisms that enhance nitrogen uptake and utilization when green manures are reintroduced into fields. Finally, the review anticipates future research directions in modulating soil nitrogen dynamics and crop nitrogen uptake through green manure application, aiming to advance research and the development of the green manure sector.

Earthquakes are one of the most harmful natural disasters to humans. Minimizing the losses caused by earthquake disasters to mankind is the goal that we have been striving to pursue. At present，earthquake prediction is still a worldwide problem.Vigorously carrying out popularization of science is one of the work measures that can effectively reduce the loss of earthquake disasters at this stage. With the developments of informatization and technology，the content and methods of popular science propaganda for earthquake prevention and disaster reduction also need to keep pace with the times in order to better adapt to the needs of economic and social development. In particular，the Yunnan earthquake system is in the period of institutional reform. How to do a good job in popularization of earthquake prevention and disaster reduction in the new era is both an opportunity and a challenge. This article takes the Kunming seismic station of Yunnan and some other stations as an example to compare with the popular science propaganda work of coastal provinces，and consider how to strengthen the popular science work of earthquake prevention and disaster reduction in Yunnan under the new situation.

Traumatic brain injury (TBI) is a serious disease that could increase the risk of epilepsy. The purpose of this article is to explore the common molecular mechanism in TBI and epilepsy with the aim of providing a theoretical basis for the prevention and treatment of post-traumatic epilepsy (PTE). Two datasets of TBI and epilepsy in the Gene Expression Omnibus (GEO) database were downloaded. Functional enrichment analysis, protein–protein interaction (PPI) network construction, and hub gene identification were performed based on the cross-talk genes of aforementioned two diseases. Another dataset was used to validate these hub genes. Moreover, the abundance of infiltrating immune cells was evaluated through Immune Cell Abundance Identifier (ImmuCellAI). The common microRNAs (miRNAs) between TBI and epilepsy were acquired via the Human microRNA Disease Database (HMDD). The overlapped genes in cross-talk genes and target genes predicted through the TargetScan were obtained to construct the common miRNAs–mRNAs network. A total of 106 cross-talk genes were screened out, including 37 upregulated and 69 downregulated genes. Through the enrichment analyses, we showed that the terms about cytokine and immunity were enriched many times, particularly interferon gamma signaling pathway. Four critical hub genes were screened out for co-expression analysis. The miRNA–mRNA network revealed that three miRNAs may affect the shared interferon-induced genes, which might have essential roles in PTE. Our study showed the potential role of interferon gamma signaling pathway in pathogenesis of PTE, which may provide a promising target for future therapeutic interventions.

Abstract Constructive remodeling of focal esophageal defects with biodegradable acellular grafts relies on the ability of host progenitor cell populations to repopulate implant regions and facilitate growth of de novo functional tissue. Intrinsic molecular mechanisms governing esophageal repair processes following biomaterial-based, surgical reconstruction is largely unknown. In the present study, we utilized mass spectrometry-based quantitative proteomics and in silico pathway evaluations to identify signaling cascades which were significantly activated during neoepithelial formation in a Sprague Dawley rat model of onlay esophagoplasty with acellular silk fibroin scaffolds. Pharmacologic inhibitor and rescue experiments revealed that epithelialization of neotissues is significantly dependent in part on pro-survival stimuli capable of suppressing caspase activity in epithelial progenitors via activation of hepatocyte growth factor receptor (c-MET), tropomyosin receptor kinase A (TrkA), phosphoinositide 3-kinase (PI3K), and protein kinase B (Akt) signaling mechanisms. These data highlight the molecular machinery involved in esophageal epithelial regeneration following surgical repair with acellular implants.

Abstract Interleukin-17 receptor D (IL17RD or IL-17RD) also known as Sef (similar expression to fibroblast growth factor), is a single pass transmembrane protein that is reported to regulate several signaling pathways . IL17RD was initially described as a feedback inhibitor of fibroblast growth factor (FGF) signaling during zebrafish and frog development. It was subsequently determined to regulate other receptor tyrosine kinase signaling cascades as well as several proinflammatory signaling pathways including Interleukin-17A (IL17A), Toll-like receptors (TLR) and Interleukin-1α (IL1α) in several vertebrate species including humans. This review will provide an overview of IL17RD regulation of signaling pathways and functions with emphasis on regulation of development and pathobiological conditions. We will also discuss gaps in our knowledge about IL17RD function to provide insight into opportunities for future investigation. Video Abstract

Expansion of visceral white adipose tissue (vWAT) occurs in response to nutrient excess, and is a risk factor for metabolic disease. SPRY1, a feedback inhibitor of receptor tyrosine kinase (RTK) signaling, is expressed in PDGFRa+ adipocyte progenitor cells (APC) in vivo. Global deficiency of Spry1 in mice results in disproportionate postnatal growth of gonadal WAT (gWAT), while iWAT and BAT were similar in size between Spry1KO and WT mice. Spry1 deficiency increased the number of PDGFRa+ stromal vascular fraction (SVF) cells in gWAT and showed increased proliferation and fibrosis. Spry1KO gWAT had increased collagen deposition and elevated expression of markers of inflammation. In vitro, SPRY1 was transiently down regulated during early adipocyte differentiation of SVF cells, with levels increasing at later stages of differentiation. SPRY1 deficiency enhances PDGF-AA and PDGF-BB induced proliferation of SVF cells. Increased proliferation of SVF from Spry1KO gWAT accompanies an increase in AKT activation. PDGF-AA stimulated a transient down regulation of SPRY1 in wild type SVF, whereas PDGF-BB stimulated a sustained down regulation of SPRY1 in wild type SVF. Collectively, our data suggest that SPRY1 is critical for regulating postnatal growth of gWAT by restraining APC proliferation and differentiation in part by regulation of PDGFRa/b-AKT signaling.

China is a country with many earthquakes，and Yunnan is a province with frequent earthquakes. The Tonghai earthquake with a magnitude of 7.8 in 1970 caused more than 10 000 casualties. In recent years，government departments have increased their efforts on propagating the science and popularization of earthquakes. The earthquake department has played an important role in earthquake prevention and mitigation. After a long period of exploration and development by earthquake people from generation to generation，the seismic station becomes the basic unit of the earthquake monitoring system. This article briefly discusses how Yunnan earthquake stations can be better expanded and used for earthquake disaster reduction and local economic development.

Pepper vein yellows virus (PeVYV) is a newly recognized Polerovirus extracted from Chinese pepper. The symptoms of PeVYV-infested pepper plants comprise intervein yellow staining, leaf curl formation and other malformations, and leaf internodal shrinkage, but the roles of the viral proteins remain undetermined. The P0 protein of the genus Polerovirus has established post-transcriptional gene silencing (PTGS) activity. This investigation focused on the PeVYV-encoded P0 protein and assessed its potential virulence capacity, PTGS activity, and tendencies to localize in the nucleus. This study revealed that P0 influenced the pathogenic properties of a specific heterologous potato virus X. In addition, P0 proteins impaired local gene silencing, although they did not regulate generalized gene silencing within Nicotiana benthamiana 16c plants. Furthermore, P0 proteins localized mainly in the nucleus, particularly in the nucleolus. P0 deletion mutagenesis demonstrated that the F-box motif (56–72 amino acids, AAs) of P0 was essential for symptom determination, inhibition of PTGS, and subcellular localization. Mutation analysis of the F-box motif of P0 protein indicated that AA 57 of the P0 protein was a pivotal site in symptom development and that AA 56 of the P0 protein was indispensable for inhibiting PTGS and subcellular localization. The outcomes obtained here suggest that further studies should be conducted on the molecular mechanisms of amino acids of the F-box domain of P0 protein in the interaction of PeVYV with plants.

Background/Aims: Circulating miRNAs could serve as biomarkers for diagnosis or prognosis of heart diseases and cerebrovascular diseases. Dexmedetomidine has protective effects in various organs. The effects of dexmedetomidine on circulating miRNAs remain unknown. Here, we investigated differentially expressed miRNA and to predict the target genes of the miRNA in patients receiving dexmedetomidine. Methods: The expression levels of circulating miRNAs of 3 patients were determined through high through-put miRNA sequencing technology. Target genes of the identified differentially expressed miRNAs were predicted using TargetScan 7.1 and miRDB v.5. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to conduct functional annotation and pathway enrichment analysis of target genes respectively. Results: Twelve differentially expressed miRNAs were identified. Five miRNAs were upregulated (hsa-miR-4508, hsa-miR-novel-chr8_87373, hsa-miR-30a-3p, hsa-miR-novel-chr16_26099, hsa-miR-4306) and seven miRNAs (hsa-miR-744-5p, hsa-miR-320a, hsa-miR-novel-chr9_90035, hsa-miR-101-3p, hsa-miR-150-5p, hsa-miR-342-3p, and hsa-miR-140-3p) were downregulated after administration of dexmedetomidine in the subjects. The target genes and pathways related to the differentially expressed miRNAs were predicted and analyzed. Conclusion: The differentially expressed miRNAs may be involved in the mechanisms of action of dexmedetomidine. Specific miRNAs, such as hsa-miR-101-3p, hsa-miR-150-5p and hsa-miR-140-3p, are new potential targets for further functional studies of dexmedetomidine.

Summary: The bladder urothelium functions as a urine-blood barrier and consists of basal, intermediate, and superficial cell populations. Reconstructive procedures such as augmentation cystoplasty and focal mucosal resection involve localized surgical damage to the bladder wall whereby focal segments of the urothelium and underlying submucosa are respectively removed or replaced and regeneration ensues. We demonstrate using lineage-tracing systems that urothelial regeneration following augmentation cystoplasty with acellular grafts exclusively depends on host keratin 5-expressing basal cells to repopulate all lineages of the de novo urothelium at implant sites. Conversely, repair of focal mucosal defects not only employs this mechanism, but in parallel host intermediate cell daughters expressing uroplakin 2 give rise to themselves and are also contributors to superficial cells in neotissues. These results highlight the diversity of urothelial regenerative responses to surgical injury and may lead to advancements in bladder tissue engineering approaches. : In this article, Mauney and colleagues show the source of urothelial progenitors utilized during bladder regeneration is dependent on the nature of injury. Fate-mapping experiments reveal that host basal and intermediate cell progeny differentially contribute to de novo urothelial formation in bladder augmentation and mucosal resection settings. These results highlight the diversity of urothelial regenerative responses to surgical injury. Keywords: bladder, tissue engineering, progenitor, urothelium

High-speed rail (HSR) is often claimed to bring different regions and cities closer together by shortening travel times, which can reduce the costs and increase enterprises productivity to promote a sustainable economy. However, another view argues that HSR transfers economic activities from peripheral cities to core cities, resulting in unbalanced regional economic development and damaging the sustainability of the economy. Based on microdata from China, this paper empirically investigates the impact of HSR on the enterprises productivity in both core cities and peripheral cities and explores the impact mechanism from the perspective of allocation effect and distribution effect caused by HSR. The results show that the connection of HSR positively affects the enterprises productivity in core cities, while it negatively affects the enterprises productivity in peripheral cities, with effect values of 1.38% and -8.45%, respectively. The conclusion still holds after endogenous treatment and multiple robustness tests are conducted. Additionally, the allocation effect analysis shows that the market access caused by HSR has an optimization effect on the resource allocation efficiency of both core cities and peripheral cities. The distribution effect analysis reveals that the distribution of enterprise productivity in peripheral cities has market heterogeneity, regional heterogeneity, and location heterogeneity. The important policy significance of this paper is that, in order to promote the sustainable development of enterprises and the economy, it should reduce policy restrictions and promote the effective flow of capital and talents, carry out the dislocation development of industry for peripheral cities, and “build a nest to attract the phoenix.”

Dielectric barrier discharge (DBD) plasma at atmospheric pressure in air was applied to a poplar veneer surface. Effects of plasma processing rate on surface morphology, chemical property, and surface wettability of the poplar veneer were investigated. The adhesion strength of urea formaldehyde (UF) glued plywood manufactured from the modified veneer was also studied. Atomic force microscopy (AFM), electron spin-resonance spectroscopy (ESR), X-ray photoelectron spectroscopy (XPS), contact angle tests, and shear strength tests were carried out. AFM indicated that the surface roughness increased after plasma treatment and was the maximum at a processing rate of 14 m/min. Both ESR and XPS tests showed more oxygen accumulation on the wood surface, forming various oxygen-containing chemical groups. Contact angle tests showed better wetting at a decreased plasma processing rate. Consequently, the adhesion strength of plywood increased after plasma treatment and showed higher strength at lower processing rates.

The effects of atmospheric pressure dielectric barrier discharge (DBD) plasma on the surface properties of wheat straw were investigated in this work. The surface wettability changes in the wheat straw were determined using contact angle measurements, the surface morphology was observed using scanning electron microscopy (SEM) and atomic force microscopy (AFM), and the chemical characteristics were scanned using X-ray photoelectron spectroscopy (XPS). Moreover, the shear strength was measured using a paper tension meter. The results indicated that after plasma treatment, the urea-formaldehyde (UF) resin had lower instantaneous and equilibrium contact angles on the wheat straw surfaces than the untreated specimens did, which decreased by 34% and 64%, respectively. Obvious etching was observed on the wheat straw surface after plasma treatment. There was an increase in the O/C ratio along with an increase in the C2, C3, and C4 proportions based on the XPS analysis after plasma treatment on wheat straw surface. Moreover, the shear strength between glued surfaces of the wheat straw was greatly improved after plasma treatment, indicating that atmospheric pressure DBD plasma treatment was an effective method for improving the surface properties of wheat straw, which were helpful for UF resin penetration and application of wheat straw.

As a transcription factor, p53 must accumulate in the nucleus to be effective. Signals related to nuclear localization are distributed mainly in the C-terminal of p53. So these nuclear location domains were reserved and the other part of the C-terminal was removed in this study. We investigated whether the truncated p53 (p53(DEL)) may affect proliferation and invasive potential of human lung cancer cells. H1299 and 801D cells expressing full-length p53 and the p53(DEL) were obtained by screening. Cell proliferation assay, cell apoptotic analysis, cell migration assay, and invasion assay were performed. Protein expression was examined by Western blotting. The data showed H1299-p53(DEL) and 801D-p53(DEL) cells grew more slowly than H1299-p53 and 801D-p53 cells, respectively. The colony formation of H1299-p53(DEL) and 801D-p53(DEL) cells reduced. The truncated p53 induced cell apoptosis. The expression levels of Bax and p53 upregulated modulator of apoptosis were increased in H1299-p53(DEL) and 801D-p53(DEL) cells. H1299-p53(DEL) and 801D-p53(DEL) cells were also characterized by decreased migration and invasion. The expression of the truncated p53 resulted in upregulation of E-cadherin and downregulation of Vimentin, Slug, Twist1, and zinc finger E-box-binding homeobox 1, which suggested the truncated p53 inhibited epithelial–mesenchymal transition occurrence. The above-mentioned characteristics were reverted by treatment of with pifithrin-a, a p53 inhibitor. These findings support the existence of a direct link between the p53(DEL), proliferation, epithelial–mesenchymal transition, and invasiveness in human lung cancer cells. So the p53(DEL) is a promising target for prevention and treatment of lung cancer.

The dynamic wettability of plasma-modified poplar veneer was investigated with sessile adhesive droplets using a wetting model. Dynamic contact angle, instantaneous and equilibrium contact angles, and their rates of change (K-value) were used to illustrate the dynamic wetting process. The experiment consisted of selecting treatment parameters (type of gas, power) that would lead to the increased wettability of wood. Three resin systems, urea-formaldehyde (UF), phenol-formaldehyde (PF), and diphenylmethylene diisocyanate (MDI), were evaluated. Based on the wetting model, the K-value was used to interpret the kinetics of wetting. The higher the K-value, the faster the contact angle reaches equilibrium, and the faster the liquid penetrates and spreads. Therefore, the model was helpful for characterizing the dynamic wettability of wood surfaces modified with different plasma treatments. The K-values of plasma-treated veneer surfaces at different plasma power levels and with different gases (such as O2, N2, Ar, air, and NH3) were 458% to 653% and 332% to 528% higher than those of untreated veneer surfaces, respectively. In addition, the K-values of the three resins on the oxygen plasma-treated veneer surfaces were 38% to 1204% higher than those on the untreated veneer surfaces. Therefore, this method was helpful for characterizing the dynamic wettability of veneer surfaces modified with plasma treatment.

The effects of cold oxygen plasma treatment on the exterior and interior surfaces and wettability of wheat straw were investigated. The wheat straw was treated with oxygen plasma for 150 s, and the radio-frequency power was set at 100 W. The surface wettability was evaluated by measuring the contact angles and the K values of urea-formaldehyde, phenol-formaldehyde, and methylene diphenyl diisocyanate resins. Specimens with different gluing surfaces were bonded together with urea-formaldehyde and phenol-formaldehyde and then hot-pressed to assess bonding strength. Results indicate that the dynamic wettability and the shear strength of wheat straw were remarkably improved after it was exposed to the cold oxygen plasma. Additionally, the adhesive type and the wheat straw surface characteristics had significant effects on the dynamic wettability and bonding strength of both untreated and plasma-treated wheat straw.

Effects of cold oxygen plasma treatment on activating the surface of poplar veneers and improving its wettability were investigated. The veneers were treated with cold oxygen plasma for 1, 3, 5, 7, and 9 min, and aged in air for 1, 3, 7, 14, 21, and 28 days. The dynamic adhesive wettability of veneers was assessed using the contact angle, K-value analysis, and surface free energy. The shear strength of three-layer panels produced from untreated and cold oxygen plasma treated veneers was examined. The results showed that the wettability of veneer was significantly improved after cold oxygen plasma treatment, leading to the enhancement of shear strength of panels. The optimized treatment time should be 7 min. Aging effect of treated veneers showed that the veneer surface wettability degraded within the first 7 days and thereafter changed slightly.

Background and objective Currently available studies on the polymorphisms of drug-metabolizing enzymes and their chemotherapeutic effects in non-small cell lung cancer are not consistent. In the present study, the relationship of the gene polymorphisms of cytochrome P450 1A1 (CYP1A1), cytochrome P450 2E1 (CYP2E1), cytochrome P450 2D6 (CYP2D6), and glutathione S-transferase M1 (GSTM1) enzymes with chemotherapeutic effects were investigated. The effects of these relationships on the survival of advanced non-small cell lung cancer patients were also examined. Methods Four drug metabolism enzymes were genotyped in lung cancer patients by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism. These patients were followed for five years. Results The chemotherapeutic effect on patients carrying B-type CYP1A1 and null-type GSTM1 was better than on those carrying other types (P0.05). Conclusion The chemotherapeutic effect on patients carrying A-type CYP1A1 was better than on those carrying the B and C types when non-platinum drugs were administered. The chemotherapeutic effect on patients carrying null-type GSTM1 was better than on those carrying the functional type when platinum drugs were administered. The four enzymes did not affect the OS of advanced non-small cell lung cancer patients.

Background and objective The p53 as a transcription factor in cell stress was activated to regulate cell cycle and programmed cell death to inhibit tumor growth. Usually, p53 is kept in non-activated state through various mechanisms, including the action of p53 C-terminal negative regulatory sequences. The purpose of the study is to prepare the two types p53 recombinant adenoviruses that carry full-length p53 as well as deletion of negative regulatory sequences at p53 C-terminus and to detect exogenous GFP expression in human lung cancer cell infected-virus by FCM scatter plot. Methods Using pAdEasy-Track vector system the p53 recombinant plasmids was constructed and the homologous recombinants in E.coli was produced. The three kinds of recombinant adenovirus in L293 cells was generated, sequencing proved. Exogenous GFP expression in human lung cancer 801D cells infectedvirus was detected by FCM scatter plot. Results p53 recombinant adenoviruses named Ad-p53(wtp), Ad-p53(del) and Ad-(empty carrier) were produced. Results of sequences indicate that the Ad-p53(del) was deletion of 111 bases before stop codon TGA and of 3 untranslated region at p53, the Ad-p53(wtp) no loss of any p53 base, the Ad-(empty carrier) no p53 sequence. FCM scatter plot indicate the percentage of 801D cells expressed GFP with three kinds of viral infection was almost same and was increased with the virus density. 801D contains ratio of cells with different fluorescence intensity. Conclusion The preparation of recombinant adenovirus, Ad-p53(del), pA-p53(wtp) and Ad-(empty carrier). The cells expressed-GFP can be quantitatively detected by FCM scatter plot. It was provide that the reliability of the virus system and accurate method for selecting viruses density to infecting cells.

Background and objective COX-2 was highly expressed in many tumor tissues and was involved in the initiation and development of tumors. The RNAi technique is a method to inhibit gene expression economically, quickly, efficiently and specifically. This study used RNAi technique to explore the interfering effect of COX-2 geneexpression and the influence on the malignant proliferation of A2 cells after quenching COX-2 in vitro . Methods Three COX-2 siRNA expression vectors with human U6 promoter were constructed. The COX-2 siRNA vectors and the vacant vector (pEGFP) were transfected into A2 cells with lipofectamine respectively. The cell strains transfected were selected. The change of COX-2 expression levels was examined by Western blot and RT-PCR. The effects on the proliferationof A2 cells after silencing COX-2 were detected by cell growth curve and clonogenic assay in vitro . Results The three siRNA and U6 promoter cloned into pEGFP plasmid were validated by PCR, restriction endonucleases identification, DNA sequencing and BLAST alignment. The cell strains transfected were coded as A2-3, A2-7, A2-10 and A2-p respectively. Green fluorescence was seen in A2-p cells and not in A2-3, A2-7 and A2-10 cells in 24 h, 48 h and 72 hafter transfected. The results of RT-PCR and Western blot showed the three siRNA expression vectors acted effectively and the expression of COX-2 was inhibited in different extent. In contrast to A2 cells, COX-2 mRNA levels of A2-3, A2-7 and A2-10 cells were reduced 15.6%, 20.4% and 64.2% respectively, and COX-2 protein expressions of them were reduced 23.7%, 36.7% and 60.2% respectively. The results of cell growth curve and clonogenic assay showed the growth of A2-10 cell slowed and the clonal formation rate was reduced. However the growth of A2-3 and A2-7 cells had no obvious changes vs controls (A2 and A2-p). Conclusion Silencing COX-2 gene in vitro by RNAi technique can significantly inhibit the malignant proliferation of A2 cells.

Background and Objective Drug resistance of lung cancer cells is one of main factors which affect the outcome of chemotherapy. It has been reported that abnormal p53 gene is well assosiated with chemotherapy resistance of tumor cells. The aim of this study is to evaluate the effects of p53 gene on drug resistance in human lung cancer celllines，so as to provide foundation of choosing individual chemotherapy drugs in clinical treatment. Methods The expression vectors which contain p53cDNA and p53 antisense cDNA respectively were constructed and were confirmed by sequencing. Transfected the 801D, a human lung cancer cell line with recombined plasmids by lipofectin mediating.Several kinds of monoclone cell lines,pEGFP-801D、pEGFP-sense p53-801D（including sense p53，pEGFP-p53(RS)-801D）、pEGFP-antisense p53-801D（including antisense p53，pEGFP-p53(AS)-801D）, which contained p53 odifferent status were obtained. Green fluorescence was observed through fluorescence microscopy. The extraneous gene was detected by PCR. MTT assay was taken to determine the drug resistance of each cell line to chemotherapy agents. Cell cycle and apoptosis induced by antitumor drugs were examined by flow cytometer. Results Extraneous sense p53 andantisense p53 were proved to be linked to plasmid respectively by sequencing.Green fluorescence was found in transfectedcell lines. The IC50 of pEGFP-p53(AS)-801D cell line（0.26±0.09 μg/mL） to Cisplatin（DDP） decreased markedly compared with 801D（0.55±0.19 μg/mL，P﹤0.05）and pEGFP-801D（0.77±0.13μg/mL，P﹤0.05）. The IC50 value of pEGFP-p53(RS)-801D to DDP is 0.43±0.25 μg/mL，which is significantly lower than that of pEGFP-801D（P ＝0.000）but higher than that of pEGFP-p53(AS)-801D(P

Changes in the vascular smooth muscle cell (VSMC) contractile phenotype occur in pathological states such as restenosis and atherosclerosis. Multiple cytokines, signaling through receptor tyrosine kinases (RTK) and PI3K/Akt and MAPK/ERK pathways, regulate these phenotypic transitions. The Spry proteins are feedback modulators of RTK signaling, but their specific roles in VSMC have not been established.Here, we report for the first time that Spry1, but not Spry4, is required for maintaining the differentiated state of human VSMC in vitro. While Spry1 is a known MAPK/ERK inhibitor in many cell types, we found that Spry1 has little effect on MAPK/ERK signaling but increases and maintains Akt activation in VSMC. Sustained Akt signaling is required for VSMC marker expression in vitro, while ERK signaling negatively modulates Akt activation and VSMC marker gene expression. Spry4, which antagonizes both MAPK/ERK and Akt signaling, suppresses VSMC differentiation marker gene expression. We show using siRNA knockdown and ChIP assays that FoxO3a, a downstream target of PI3K/Akt signaling, represses myocardin promoter activity, and that Spry1 increases, while Spry4 decreases myocardin mRNA levels.Together, these data indicate that Spry1 and Spry4 have opposing roles in VSMC phenotypic modulation, and Spry1 maintains the VSMC differentiation phenotype in vitro in part through an Akt/FoxO/myocardin pathway.

Development of the hematopoietic and endothelial lineages derives from a common mesodermal precursor, the Flk1(+) hemangioblast. However, the signaling pathways that regulate the development of hematopoietic and endothelial cells from this common progenitor cell remains incompletely understood. Using mouse models with a conditional Spry1 transgene, and a Spry1 knockout mouse, we investigated the role of Spry1 in the development of the endothelial and hematopoietic lineages during development.Quantitative RT-PCR analysis demonstrates that Spry1, Spry2, and Spry4 are expressed in Flk1(+) hemangioblasts in vivo, and decline significantly in c-Kit(+) and CD41(+) hematopoietic progenitors, while expression is maintained in developing endothelial cells. Tie2-Cre-mediated over-expression of Spry1 results in embryonic lethality. At E9.5 Spry1;Tie2-Cre embryos show near normal endothelial cell development and vessel patterning but have reduced hematopoiesis. FACS analysis shows a reduction of primitive hematopoietic progenitors and erythroblastic cells in Spry1;Tie2-Cre embryos compared to controls. Colony forming assays confirm the hematopoietic defects in Spry1;Tie2-Cre transgenic embryos. Immunostaining shows a significant reduction of CD41 or CD71 and dpERK co-stained cells in Spry1;Tie2-Cre embryos compared to controls, whereas the number of VEC(+) and dpERK co-stained cells is comparable. Compared to controls, Spry1;Tie2-Cre embryos also show a decrease in proliferation and an increase in apoptosis. Furthermore, loss of Spry1 results in an increase of CD41(+) and CD71(+) cells at E9.5 compared with controls.These data indicate that primitive hematopoietic cells derive from Tie2-expressing hemangioblasts and that Spry1 over expression inhibits primitive hematopoietic progenitor and erythroblastic cell development and expansion while having no obvious effect on endothelial cell development.