Transforming growth factor-beta 1 enhances discharge activity of cortical neurons

Transforming growth factor-beta 1(TGF-β1)has been extensively studied for its pleiotropic effects on central nervous system diseases.The neuroprotective or neurotoxic effects of TGF-β1 in specific brain areas may depend on the pathological process and cell types involved.Voltage-gated sodium channels(VGSCs)are essential ion channels for the generation of action potentials in neurons,and are involved in various neuroexcitation-related diseases.However,the effects of TGF-β1 on the functional properties of VGSCs and firing properties in cortical neurons remain unclear.In this study,we investigated the effects of TGF-β1 on VGSC function and firing properties in primary cortical neurons from mice.We found that TGF-β1 increased VGSC current density in a dose-and time-dependent manner,which was attributable to the upregulation of Nav1.3 expression.Increased VGSC current density and Nav1.3 expression were significantly abolished by preincubation with inhibitors of mitogen-activated protein kinase kinase(PD98059),p38 mitogen-activated protein kinase(SB203580),and Jun NH2-terminal kinase 1/2 inhibitor(SP600125).Interestingly,TGF-β1 significantly increased the firing threshold of action potentials but did not change their firing rate in cortical neurons.These findings suggest that TGF-β1 can increase Nav1.3 expression through activation of the ERK1/2-JNK-MAPK pathway,which leads to a decrease in the firing threshold of action potentials in cortical neurons under pathological conditions.Thus,this contributes to the occurrence and progression of neuroexcitatory-related diseases of the central nervous system.

Microbiological Transformation of Ginsenoside Rg_1

Forty-nine microbial strains were used to screen their ability for the microbiological transforma-tion of ginsenoside Rg1. Aspergillus niger (3.1858) and Absidia coerulea (3.3538) were found to convert ginsenoside Rg1 efficiently to less polar metabolites. Preparative scale transformation with both fungi Absidia coerulea (3.3538) and Aspergillus niger (3.1858) have resulted in the production of one same metabolite (MT1). Its structure was char-acterized as 6-O-b-D-glucopyranosyl-20(S)-protopanaxatriol (Ginsenoside Rh1) on the basis of its TOF-MS and 1H, 13C NMR spectral data. The biotransformation kinetic curves for Ginsenoside Rg1 and MT1 were reported for the first time, and the biotransformation pathway was proposed.

Studies on Genetic Transformation of NPR1 Gene into Maize by Microprojectile Bombardment

[Objective] This study aimed to explore the conditions of transformation of maize by microprojectile bombardment. [Method] Immature embryo-derived callus of maize inbred line 7239 was used as explants to study the effects of shoot distance, helium pressure, vacuum and bombardment frequency on the transformation efficien- cy in the particle bombardment system of maize. [Result] Considering the transfor- mation efficiency, particle bombardment with 100 μg/P of golden particles, at a shoot distance of 9 cm from the target cells, under helium pressure of 1 350 psi and vac- uum 25 inHg, and bombarding twice could achieve relatively ideal results. After se- lection on media supplemented with different concentration of hygromycin, some re- generated plants were obtained. The results of PCR and Southern blotting analysis demonstrated that the NPR1 gene had been integrated into the genome of trans- genic maize plants, with an average transformation efficiency of 1.76%. [Conclusion] The study laid the foundation for the cultivation and breeding of excellent resistant varieties of maize.

Cloning and Gentic Transformation of NbDAD1 Gene from Nicotiana benthamiana

[Objective] The aim was to clone NbDAD1 gene from Nicotiana benthami- ana and study its genetic transformation. [Method] NbDAD1 gene was isolated from N. benthamiana by using RT-PCR technology and over-expression vectors were con- structed to obtain NbDADl-overexpression resistant plants and NbDADl-overexpres- sion resistant plants carrying HA tag. [Result] The 351 bp long NbDAD1 gene was cloned from N. benthamiana; recombinant plasmids pCAMBIA1301-NbDAD1 and pCAMBIA1301-NbDAD1HAtag were constructed successfully; 50T0-generation N. ben- thamiana Hyg-resistant transgenic lines of three genotypes were obtained, including 23 positive transgenic plants. [Conclusion] This study laid the foundation for investi- gating the specific functions of NbDAD1 gene in N. benthamiana and exploring the possible functional mechanism of DAD1 protein in programmed cell death of plants.

Construction and Genetic Transformation of Over-Expression Vector of nad1 Gene in Rice

[Objective] The aim was to clone the mitochondrial-related gene nad1 and produce transgenic rice plants with nad1.[Method] The total RNA was extracted from rice seedlings and reverse transcripted into cDNA.Then the target gene nad1 was amplified by using the cDNA as template.The nad1 and Rf1b,a sequence of signal peptide of mitochondria,were linked to binary expression vector pCAMBIA1305.1.The recombinant plasmid was transformed into the callus by Agrobacterium-mediated approach.[Result] The target gene nad1 was 978 bp.The binary expression vector carrying nad1 and signal peptide of mitochondria was constructed successfully.In addition,a lot of transgenic plants were obtained.[Conclusion] The study will provide basis to investigate the effect of over-expression of nad1 on rice plant growth.

TGF-β1-promoted epithelial-to-mesenchymal transformation and cell adhesion contribute to TGF-β1-enhanced cell migration in SMMC-7721 cells

Transforming growth factor-b 1 (TGF-β1), a multi-function polypeptide, is a double-edged sword in cancer. For some tumor cells, TGF-β1 is a potent growth inhibitor and apoptosis inducer. More commonly, TGF-β1 loses its growth-inhibitory and apoptosis-inducing effects, but stimulates the metastatic capacity of tumor cells. It is currently little known about TGF-β1-promoted cell migration in hepatocellular carcinoma (HCC) cells, let alone its mechanism. In this study, we found that TGF-β1 lost its tumor-suppressive effects, but significantly stimulated cell migration in SMMC-7721 human HCC cells. By FACS and Western blot analysis, we observed that TGF-β1 enhanced the expression of α5β1 integrin obviously, and subsequently stimulated cell adhesion onto fibronectin (Fn). Furthermore, we observed that TGF-β1 could also promote SMMC-7721 cells adhesion onto laminin (Ln). Our data also provided evidences that TGF-β1 induced epithelial-to-mesenchymal transformation (EMT) in SMMC-7721 cells. First, SMMC-7721 cells clearly switched to the spindle shape morphology after TGF-β1 treatment. Furthermore, TGF-β1 induced the down-regulation of E-cadherin and the nuclear translocation of β-catenin. These results indicated that TGF-β1-promoted cell adhesion and TGF-β1-induced epithelial-to-mesenchymal transformation might be both responsible for TGF-β1-enhanced cell migration.

Expression transformation of claudin-1 in the process of gastric adenocarcinoma invasion

AIM: To investigate the relation of expression transfor-mation of claudin-1 with invasiveness and metastasis of gastric carcinoma. METHODS: By using immunohistochemistry, expres-sion of claudin-1 in mucosa and invasive front of 136 gastric adenocarcinoma cases and proliferative index (Ki-67) were detected and analyzed. RESULTS: In mucosa, the claudin-1 over-expression rate of mucinous adenocarcinomas (including signet-ring cell carcinomas) was the highest. It was nega-tively related with the differentiation but positively related with the invasiveness and metastasis of gastric cancer. In invasive front, the claudin-1 over-expression rate was positively related with the differentiation, in-vasiveness and metastasis of gastric carcinoma. The expression transformation of claudin-1 was found in gastric carcinoma. The expression of claudin-1 in inva-sive front was transformed in 28/136 gastric carcinoma cases. The transformation rate in highly differentiated tubular adenocarcinomas was the highest (51.5%, 17/33). The deeper was the invasiveness, the higher was the transformation rate. The claudin-1 expression transformation rate in serosa and omenta was signifi -cantly higher (92.9%) than in tunica muscularis of in-vasive gastric cancer cases, as well as in patients withlymph node metastasis than in those without lymph node metastasis. CONCLUSION: Up-regulation of claudin-1 expres-sion and its transformation in invasive and metastatic gastric carcinoma suggest that claudin-1 participates in the transformation of biological behaviors in neo-plasms. Further study is needed to elucidate the pre-cise mechanism and the relation of claudin-1 expres-sion with the neoplasm progress.

Darboux Transformations for Space-Like Isothermic Surfaces in R^m,1

In this paper,by using the G_(m,1)~(1,1)-system,we study Darboux transformations for space-like isothermic surfaces in Minkowski space R~(m,1),where G_(m,1)~(1,1)=O(m+1,2)/O(m,1)×O(1,1).

AKT1 and AKT2 Promote Malignant Transformation in Human Brain Glioma LN229 Cells

OBJECTIVE To confirm the role played by AKT1 and AKT2 in the β- catenin/Tcf-4 signaling pathway in promoting malignant transfor- mation of glioma cells. METHODS LN229 cells were divided into five groups： a control group, acetone （ACE）group, acetylsalicylic acid （ASA; aspirin） group, ASA＋AKT1 plasmid group and ASA＋AKT2 plasmid group. Western blot and PCR were used to detect the expression of AKT1 and AKT2 after dealing with ASA and transferring AKTI/2 genes into LN229 cells. Cell proliferation was determined by flow cytometry, cell invasion was evaluated by transwell assay and cell apoptosis was detected with annexin V staining. The molecules regulating proliferation and invasion were examined by western blot analysis. RESULTS Aspirin down-regulates AKT1 and AKT2 expression by modulating β-cateninfrcf-4 activity. AKT1 and AKT2 can enhance cell proliferation and invasion by up-regulating the expression of cyclin-D and matrix metalloprotein-9 （MMP-9） in LN229 glioma cells. CONCLUSION AKT1 and AKT2 play an important role in the β- catenin/Tcf-4 signaling pathway promoting malignant transformation; AKT1 is more effective than AKT2. AKT1 and AKT2 may be potential targets for brain glioma therapy and an effective way to prevent metastasis of gliomas.

Alterative Expression and Sequence of Human Elongation Factor-1δ during Malignant Transformation of Human Bronchial Epithelial Cells Induced by Cadmium Chloride

Objective To study the alternative expression and sequence of human elongation factor-1δ （human EF-1δ p31） during malignant transformation of human bronchial epithelial cells induced by cadmium chloride （CdCl2） and its possible mechanism. Methods Total RNA was isolated at different stages of transformed human bronchial epithelial cells （16HBE） induced by CdCl2 at a concentration of 5.0 μM. Special primers and probe for human EF-1δ p31 were designed and expression of human EF-18 mRNA from different cell lines was detected with fluorescent quantitative PCR technique. EF-18 cDNA from different cell lines was purified and cloned into pMD 18-T vector followed by confirming and sequencing analysis. Results The expressions of human EF-1δ p31 at different stages of 16HBE cells transformed by CdCl2 was elevated （P〈0.01 or P〈0.05）. Compared with their corresponding non-transformed ceils, the overexpression level of EF-15 p31 was averagely increased 2.9 folds in Cd-pretransformed cells, 4.3 folds in Cd-transformed ceils and 7.2 folds in Cd-tumorigenic cells. No change was found in the sequence of overexpressed EF-1δ p31 at different stages of 16HBE cells transformed by CdCl2. Conclusion Overexpression of human EF-1δ p31 is positively correlated with malignant transformation of 16HBE cells induced by CdCl2, but is not correlated with DNA mutations.

Darboux Transformation and New Soliton-like Solutions for （1＋1）-Dimensional Higher-Order Broer-Kaup System

In this letter, we construct a kind of new Darboux transformation for the （1＋1）-dimensional higher-order Broer-Kaup （HBK） system with the help of a gauge transformation of a spectral problem. By applying this new Darboux transformation, some new soliton-like solutions of the （1＋1）-dimensional HBK system are obtained.

Transforming growth factor-β1 and vascular endothelial growth factor levels in senile acute myeloid leukemia and correlation with prognosis

BACKGROUND Acute myeloid leukemia(AML)is a disease in which immature hematopoietic cells accumulate in the bone marrow and continuously expand,inhibiting hematopoiesis.The treatment and prognosis of this disease have always been unsatisfactory.AIM To investigate the correlation between vascular endothelial growth factor(VEGF)and transforming growth factor-β1(TGFβ1)expression and prognosis in older adults with AML.METHODS This study enrolled 80 patients with AML(AML group),including 36 with complete response(AML-CR),23 with partial response(AML-PR),and 21 with no response(AML-NR).The expression levels of VEGF and TGFβ1 were detected by reverse transcription polymerase chain reaction in bone marrow mononuclear cells isolated from 56 healthy controls.Kaplan-Meier analysis was performed to assess overall survival(OS)and progression-or disease-free survival(DFS).Prognostic risk factors were analyzed using a Cox proportional hazards model.RESULTS The AML group showed a VEGF level of 2.68±0.16.VEGF expression was lower in patients with AML-CR than those with AML-PR or AML-NR(P<0.05).TGFβ1 expression in the AML group was 0.33±0.05.Patients with AML-CR showed a higher TGFβ1 expression than those with AML-PR or AML-NR(P<0.05).VEGF and TGFβ1 expression in patients with AML was significantly correlated with the counts of leukocytes,platelets,hemoglobin,and peripheral blood immature cells(P<0.05);Kaplan-Meier survival analysis revealed that patients with high TGFβ1 expression had better OS and DFS than those with low TGFβ1 expression(P<0.05),whereas patients with low VEGF levels showed better OS and DFS than those with high VEGF levels(P<0.05).VEGF,TGFβ1,and platelet count were identified by the Cox proportional hazards model as independent risk factors for OS(P<0.05),while VEGF,TGFβ1,and white blood cell count were independent risk factors for DFS(P<0.05).CONCLUSION Decreased VEGF expression and increased TGFβ1 expression in patients with AML provide valuable references for determining and individualizing clinical treatment strategies.

A Discussion on the Characteristics of Tectonic Transformation and Its Mechanism of 2nd Episode of Zhu-Qiong Movement in Zhu 1 Depression, South China

Based on the detailed interpretation of high-accuracy 3D seismic data, the characteristics of regional unconformities, fracture systems, sedimentary filling and paleo-stress field in Zhu 1 depression were comprehensively analyzed. The results showed that 2nd episode of Zhu-Qiong movement was an important tectonic transformation that occurred during rifting, basin structure patterns above and below the regional angular unconformity caused by 2nd episode had distinct differences. The orientations of those basin-controlling faults changed from NE, NEE to EW, NWW, which indicated that the paleo-stress field veered from NNW extension clockwise to sub-NS extension and basin structures patterns transformed from NE, NEE strong rifting to sub-EW, NWW weak rifting. The depocenter showed seesaw-like migration from S to N. And the sedimentary systems transformed from semi-deep-deep lacustrine facies to braided river delta facies developed in shore-shallow lacustrine. Combined with its tectonic recombination of periphery plates that period, the mechanism of above tectonic transformation had direct correspondence to the readjustment of the Indian plate and the Pacific plate. The southward subduction of the pro-to-south China Sea is the direct cause of this tectonic transformation.

On an Auto-Baecklund Transformation for (2+1)-Dimensional VariableCoefficient Generalized KP Equations and Exact Solutions

By the application of the extended homogeneous balance method, we derive anauto-Backlund transformation (BT) for (2+1)-dimensional variable coefficient generalized KPequations. Based on the BT, in which there are two homogeneity equations to be solved, we obtainsome exact solutions containing single solitary waves.

Studies on Genetic Transformation of Fresh-cut Chrysanthemum Using DREB1A Promoted by Stress-induced Promoter rd29A

In this study, DERB1A transcription factor and stress-induced promoter rd29A were isolated respectively and amplified from Arabidopsis thaliana, se- quenced and analyzed by DNAsis. In addition, the stress-induced promoter rd29A was utilized to construct the plant expression vector of DERB1A, which was transformed into Agrobacterium tumefaciens. Furthermore, the transgenic regeneration system of fresh-cut chrysanthemum from callus to plantlets was established successfully. On this basis, chrysanthemum leaf-disc explants were genetically transformed with Agrobacterium-mediated method. Two positive transgenie plantlets were obtained in vitro. Based on PCR detection, DREB1A transcription factor was integrated into chrysanthemum genome, which laid the foundation for breeding new transgenie cultivars of fresh-cut chrysanthemum with high comprehensive stress resistance, good cmalitv and high field.

Studies on Cloning and Transformation of CBF1 Gene of Maize Grass

[Objective]It is revealed whether the similar maize transcriptional activator in CBF1 gene is regulatory cold resistance gene to lay the foundation for breeding new transgenic Forage Maize Varieties with high cold resistance ability.[Methods]In the present paper,the transcriptional factor gene CBF1 was Successfully cloned by PCR from the leaves of Arabidopsis.The sequence was preliminarily analyzed and plant expression vector was constructed.Then with agrobacterium-mediated transgene technique,CBF1 gene was introduced into maize SAUMZ1.[Results]PCR assay revealed that the CBF1 gene was integrated in the maize grass SAUMZ1 genome.Under different low temperature treatment,the relative electrolyte leakage percentage of transgenic plant was lower than Control.[Conclusion] The results showed that the cold-resistance of maize grass SAUMZ1 enhanced after transforming CBF1 gene.

Transient Expression of VHB and GLUT1 in the Green Alga Haematococcus pluvialis Using Agrobacteriummediated Transformation

The first successful Agrobacterium-mediated transformation of the green alga Haematococcus pluvialis using the vectors hosting the genes coding for VHB （ Vitreoscilla hemoglobin） and GLUT1 （glucose transporter 1 ） is reported here. The greenish yellow fluorescence of EGFP was observed when the zeocin-resistant cells were viewed with a fluorescent microscope. The functional expression of EGFP showed that the Agrobacterium-mediated transformation could efficiently transfer the exogenous gene into H. pluvialis. RT-PCR was used to successfully amplify the mRNA of VHB and GLUT1 genes from transformed cells, while Southern blots indicated the integration of VHB and GLUT1 genes into the genome of H. pluvialis. Transferring VHB and GLUT1 genes into this alga would pave the way for manip- ulation of many important pathways relevant to the food, pharmaceutical, and nutraceutical industries.

Intramolecular Oxidative Transformations of 1，4－Diary1－2， 3－Dimethy1－2， 3－Epoxybutane with DDQ

Treatment of 1.4-diary1 -2. 3 -dimethy1-2, 3-epoxide of butane(1) with 2. 3-dichloro-5.6 6dicyano-1.4-benzoquinone(DDQ) in trifluoroacetic acid (TFA)furnishes a new benzofuran typeIignan (2) The structure of (2) was esbolished by MS,IR,UV and NMR spectra.and X-ravcrystallographic analysis.

UV Light Induced Transformation of 1-Methylnaphthalene in the Presence of Air and Its Implications for Contaminants Research

Understanding chemical transformations of contaminants and the resulting products is extremely important in devising proper monitoring methods for such contaminants and in assessing potential human exposure to the transformation products in the environment. Ultraviolet (UV) light from the sun can induce various photochemical transformations of contaminants in the environment. Alkylnaphthalenes are light-molecular-weight polycyclic aromatic hydrocarbons (PAHs) which are one of the most widespread organic pollutants present in ambient air as a result of a variety of incomplete combustion sources. In this study, 1-methylnapthalene,a typical example of an alkylnaphthalene, was subjected to UV irradiation to investigate its transformation in the presence and absence of air. Twenty-one products were detected in the reaction mixtures. Some photo-oxidation products were identified, including both ring-opened and ring-retained oxygenated compounds, such as 1-naphthaldehyde, 1-naphthoic acid, 1-naphthalenemethanol and phthalic anhydride. Although dimeric products were observed in the presence of air, more were found in the presence of helium or argon gas, indicating a different photo-oxidation pathway from those commonly observed in other media, such as water. Under just 48 hours of exposure to the UV light in the presence of air, three major products were formed with a production yield of about 10% each. Compared to 1-methylnapthalene, the UV induced transformation products observed in this study are more volatile, acidic, water soluble or toxic. The formation of these products may significantly change our understanding of the risks assessed solely from the parent compound in contaminants research and supports the inclusion of airborne transformations of the parent compound in risk assessment.

缺氧微环境下HIF-1α通过PD-L1调控肝癌恶性生物学行为和上皮间质转化的机制研究

目的探讨缺氧微环境下,缺氧诱导因子-1α(HIF-1α)通过程序性死亡配体1(PD-L1)调控肝癌细胞增殖、迁移、侵袭和上皮间质转化的机制。方法选取2016年9月至2018年9月在徐州医科大学附属盐城临床学院确诊治疗的57例肝癌病人作为研究对象。免疫组织化学检测HIF-1α、PD-L1的表达水平,统计学分析HIF-1α、PD-L1与肝癌病人临床病理及总生存期之间的关系。通过氯化钴建立肝癌细胞缺氧模型,用细胞计数试剂盒(cell counting kit-8,CCK-8)法检测不同浓度氯化钴作用下细胞存活率;划痕实验、Transwell实验检测缺氧细胞及亲本细胞迁移、侵袭能力的变化;蛋白质印迹法检测缺氧细胞及亲本细胞HIF-1α、PD-L1蛋白表达水平。设计针对HIF-1α的siRNA(小干扰RNA,small interfering RNA)及PD-L1质粒,脂质体转染肝癌细胞氯化钴缺氧细胞,划痕实验、Transwell侵袭实验检测转染前后细胞迁移、侵袭能力的变化;蛋白质印迹法检测转染前后细胞HIF-1α、PD-L1、上皮间质转化(EMT)相关蛋白表达水平。结果与癌旁组织相比,HIF-1α、PD-L1在肝癌组织中高表达(P<0.05)。肝细胞癌中HIF-1α高表达组与低表达组在肿瘤长径、肿瘤分化程度和CNLC分期差异有统计学意义(P<0.05);PD-L1高表达组与低表达组在肿瘤数目、AFP表达水平和CNLC分期差异有统计学意义(P<0.05);且log-rank检验方法显示:HIF-1α、PD-L1高表达组病人的总生存期较低表达组明显降低(P<0.05)。与常氧细胞相比,缺氧细胞穿透小室的细胞数显著增加[(429.33±12.01)个比(244.3±8.14)个,t=−22.08,P<0.001],氯化钴缺氧肝癌细胞增殖、迁移及侵袭能力增强,HIF-1α、PD-L1表达水平明显增加(P<0.05)。与亲本细胞相比,HIF-1αsiRNA作用后肝癌缺氧细胞增殖、迁移及侵袭能力明显降低(P<0.05),且PD-L1、波形蛋白(Vimentin)及扭曲相关蛋白1(Twist1)蛋白表达明显降低,而上皮钙黏素(E-cadherin)蛋白表达明显增加(P<0.05);HIF-1αsiRNA和PD-L1质粒共转染后,可逆转HIF-1αsiRNA介导的侵袭、迁移能力的改变及EMT相关指标的改变。结论HIF-1α、PD-L1在肝癌中高表达,两者高表达预示肝癌病人预后较差。缺氧微环境下,HIF-1α可通过PD-L1调控肝癌恶性生物学行为及上皮间质转化进程。