MiR-183-5p promotes the progression of non-small cell lung cancer through targeted regulation of FOXO1

Objective To investigate miR-183-5p targeting to forkhead box protein O1(FOXO1)and its corresponding effect on the proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT)of non-small cell lung cancer(NSCLC)cells.Methods NSCLC tissues and adjacent normal tissues from 60 patients with NSCLC adenocarcinoma were obtained via pathological biopsy or intraoperative resection.Several cell lines were cultured in vitro,including the human normal lung epithelial cell line BEAS-2B and human NSCLC cell lines A549,SPCA-1,PC-9,and 95-D.miR-183-5p and FOXO1 mRNA expression in tissues and cells were detected by qRT-PCR;the corresponding correlations in NSCLC tissues were analyzed using the Pearson test,and the relationship between miR-183-5p expression and clinicopathological parameters was analyzed.The miR-183-5p-mediated regulation of FOXO1 was verified by bioinformatics prediction alongside double luciferase,RNA-binding protein immunoprecipitation(RIP)assay,and pull-down experiments.A549 cells were divided into control,anti-miR-NC,anti-miR-183-5p,miR-NC,miR-183-5p,miR-183-5p+pcDNA3.1,and miR-183-5p+pcDNA3.1-FOXO1 groups.Cell proliferation,invasion,migration,apoptosis,and cell cycle distribution were detected using an MTT assay,clone formation assay,Transwell assay,scratch test,and flow cytometry,respectively.The expression of EMT-related proteins in the cells was analyzed by western blotting.The effect of miR-185-3p silencing on the development of transplanted tumors was detected by analyzing tumor formation in nude mice.Results miR-183-5p expression was significantly higher in NSCLC tissues and cells than in adjacent normal tissues,whereas FOXO1 mRNA expression was significantly down-regulated.There was a significant negative correlation between miR-183-5p and FOXO1 mRNA in NSCLC tissues(P<0.05).Additionally,the expression of miR-183-5p was significantly correlated with tumor size,tumor differentiation,and tumor-node-metastasis stage in patients with NSCLC(P<0.05).miR-183-5p targeted and inhibited FOXO1 expression.Compared to the anti-miR-NC group,the cell proliferation,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells were significantly lower in the anti-miR-183-5p group,whereas the protein expression of E-cadherin andα-catenin and the proportion of G0/G1 phase cells were significantly higher;additionally,the frequency of colony formation and invasion were significantly lower in the anti-miR-183-5p group(P<0.05).Compared to the miR-NC group,the cell proliferation,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells in the miR-183-5p group were significantly higher,whereas the E-cadherin andα-catenin protein expression and the proportion of G0/G1 phase cells were significantly lower;furthermore,the frequency of colony formation and invasion were significantly higher in the miR-183-5p group(P<0.05).Compared with the miR-183-5p+pcDNA3.1 group,the OD value,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells were significantly lower in the miR-183-5p+pcDNA3.1-FOXO1 group,whereas E-cadherin andα-catenin protein expression and the proportion of G0/G1 phase cells were significantly higher;additionally,the frequency of colony formation and invasion was significantly lower in the miR-183-5p+pcDNA3.1-FOXO1 group(P<0.05).Overall,silencing miR-185-3p inhibited the growth of transplanted tumors and promoted FOXO1 expression.Conclusion Overexpression of miR-183-5p can inhibit apoptosis and promote the proliferation,migration,invasion,and EMT,of NSCLC cells by down-regulating FOXO1 expression.

Immunoregulatory Effects of Ethyl-acetate Fraction of Extracts from Tetrastigma Hemsleyanum Diels et. Gilg on Immune Functions of ICR Mice

Objective To evaluate the effects of ethyl-acetate fraction （EAF） of extracts from Tetrastigma hemsleyanum Diels et. Gilg （TDG） on immune functions of ICR mice. Methods ICR mice were exposed to different doses of EAF for 15 or 30 days and then their immune functions were analyzed, including ConA-induced splenic lymphocyte transformation, SRBC- induced delayed type hypersensitivity response, serum hemolysin analysis, antibody-producing cells, peritoneal macrophage phagocytized chicken red blood cells, natural killer cell activity, and serum level of cytoldnes. Results EAF of extracts from TDG at different doses had various effects on immune functions of ICR mice. As compared with the controls, it increased the mouse spleen lymphocyte transformation induced by ConA, the left-hind voix pedis thickness and the number of plague forming cells （PFCs） at the dose of 1.82 mg/mL, 5.48 mg/mL, and 9.12 mg/mL, respectively; increased the ink clearance ability at the dose of 0.91 mg/mL, 1.82 mg/mL, 5.48 mg/mL, and 9.12 mg/mL, respectively; increased the phagocytosis index of mononuclear-macrophages and production of serum interferon-gamma （IFN-？） at the dose of 5.48 mg/mL; and could promote the production of serum tumor necrosis factor-alpha （TNF-α） at the dose of 9.12 mg/mL. Conclusion EAF of extracts from TDG can regulate mouse immune functions in vivo.

Sheng Jiang San, traditional multi-herb formulation, exerts anti-influenza effects in vitro and in vivo via neuraminidaseinhibition and immune regulation

OBJECTIVE Sheng Jiang San(SJS),a multi-herb formulation,is used in treating high fever,thirsty and anxiety in ancient China and it is sometimes used to treat seasonal influenza in modern.However,there is no evidencebased investigation and mechanism research to support SJS′s anti-influenza efficacy.This study aims to investigate the anti-influenza effect of SJS and its possible mechanisms.METHODS In this study,we examined the inhibitory effect of SJS against different influenza viruses on Madin-Darby canine kidney cells.Influenza virus infected BALB/c mice were employed as in vivo model to evaluate the efficacy.Mice challenged with A/PR/8/34(H1N1)were orally administrated SJS 1 g·kg^-1 daily for seven days and monitored for 14 d.The survival rate,body mass changes,lung index,lung viral load,histopathologic changes and immune-regulation of the mice were measured.The underlying anti-influenza virus mechanisms were studied by a series of biological assays in vitro to determine if hemagglutinin,ribonucleoprotein complex or nerauminidase were targets of SJS.RESULTS SJS exerted a broad spectrum of inhibitory effects on multiple influenza strains in a dose-dependent manner.And IC50 of SJS against A/WSN/33(H1N1)was lower than 35 mg·L^-1.SJS also protected 50%of mice from influenza virus PR8 infection.The lung index and the lung viral load of SJS treated mice were signifi⁃cantly decrease compared with untreated mice.SJS 2 g·L^-1 inhibited 80%of neuraminidase enzymatic activity.SJS also up-regulated TNF-αand IFN-αand down-regulated IL-2 of influenza virus induced mice.CONCLUSION SJS is a useful formulation for treating influenza virus infection.

TLR4/9介导牙周炎宿主CD25+B细胞表达IL-10、IL-35及TGF-β的效应及机制

目的分析牙周炎宿主CD25^(+)B细胞中IL-10、IL-35及TGF-β的表达特点,评价TLR4/9的表达活化对上述过程的影响。方法SD大鼠随机分为健康组、早期牙周炎组、晚期牙周炎组,采用结扎法建立牙周炎模型。检测各组动物牙龈及外周血CD25^(+)B细胞内IL-10、IL-35和TGF-βmRNA的表达水平,牙龈CD25^(+)B细胞内TLR 2/4/7/9、MyD88、TRAF6的表达活化水平。建立细胞培养体系,分析TLRs/MyD88信号对CD25^(+)B细胞表达分泌IL-10、IL-35和TGF-β的影响。结果早期牙周炎组牙龈CD25^(+)B细胞内IL-10、TGF-βmRNA表达水平高于健康组(P<0.05)。晚期牙周炎组牙龈CD25^(+)B细胞内IL-10、IL-35及TGF-βmRNA表达水平高于健康组(P<0.05),外周血CD25^(+)B细胞内IL-10 mRNA高于健康组(P<0.05),余差异无统计学意义(P>0.05)。相比健康组,晚期牙周炎组牙龈CD25^(+)B细胞内TLR4/9及MyD88的表达磷酸化水平均升高(P<0.01)。细胞实验结果显示,TLR4激动剂上调IL-10、IL-35、TGF-βmRNA表达及IL-10的浓度(P<0.05),TLR9激动剂上调IL-10、TGF-βmRNA表达及IL-10的浓度(P<0.05),TLR4/TLR9激动剂联合应用上调所有检测指标的表达及浓度(P<0.05),MyD88拮抗抑制上述过程(P<0.05)。结论牙周炎晚期,牙龈CD25^(+)B细胞表达IL-10、IL-35和TGF-β水平升高,这一过程可能受TLR4/9-MyD88信号调控。

Physiological roles of mitogen-activated-protein-kinase-activated p38-regulated/activated protein kinase

Mitogen-activated protein kinases(MAPKs)are a family of proteins that constitute signaling pathways involved in processes that control gene expression,cell division, cell survival,apoptosis,metabolism,differentiation and motility.The MAPK pathways can be divided into conventional and atypical MAPK pathways.The first group converts a signal into a cellular response through a relay of three consecutive phosphorylation events exerted by MAPK kinase kinases,MAPK kinase,and MAPK.Atypical MAPK pathways are not organized into this three-tiered cascade.MAPK that belongs to both conventional and atypical MAPK pathways can phosphorylate both non-protein kinase substrates and other protein kinases.The latter are referred to as MAPK-activated protein kinases.This review focuses on one such MAPK-activated protein kinase,MAPK-activated protein kinase 5(MK5)or p38-regulated/activated protein kinase(PRAK).This protein is highly conserved throughout the animal kingdom and seems to be the target of both conventional and atypical MAPK pathways.Recent findings on the regulation of the activity and subcellular localization,bona fide interaction partners and physiological roles of MK5/PRAK are discussed.

Long noncoding RNA negative regulator of antiviral response contributes to pancreatic ductal adenocarcinoma progression via targeting miR-299-3p

BACKGROUND Pancreatic ductal cancer(PDAC)has high malignancy and poor prognosis.Long noncoding RNAs(lncRNAs)are associated with high levels of malignancy,including PDAC.However,the biological and clinical significance of negative regulator of antiviral response(NRAV)in PDAC is unclear.AIM To study the regulatory role of lncRNA NRAV in PDAC.METHODS GEPIA analyzed lncRNA NRAV and miRNA(miR-299-3p)expression levels in PDAC tissues and measured them in PDAC cells by quantitative measurements in real time.The specific role of NRAV and miR-299-3p in cell proliferation and transfer potential was evaluated by cell formation analysis,Cell Counting Kit-8 and Transwell analysis.The relationship between NRAV and miR-299-3p was studied by predictive bioinformatics,RNA immunoassay,and fluorescence enzyme analysis.In vivo experiments included transplantation of simulated tumor cells under naked mice.RESULTS The expression level of lncRNA NRAV was higher in both tumor tissues and cell lines of PDAC and was negatively associated with the clinical survival of PDAC patients.Functionally,overexpression of NRAV promoted cell proliferation and metastasis of PDAC cells,while knockdown of NRAV reversed these effects.Finally,NRAV was performed as a molecular sponge of miR-299-3p.Moreover,overexpression of miR-299-3p could reverse the promoting effects of NRAV on cell proliferation and metastasis of PDAC cells.CONCLUSION NRAV facilitates progression of PDAC as a molecular sponge of miR-299-3p and may be a potential molecular marker for diagnosis and treatment of PDAC.

Synthesis,Crystal Structure and Plant Growth Regulatory Activity of 1-(3-Amino-[1,2,4]triazol-1-yl)-3,3-dimethyl-butan-2-one

The title compound 1-（3-amino-[1,2,4]triazol-1-yl）-3,3-dimethyl-butan-2-one（3） was synthesized by Hofmann-alkylation reaction of 1-chloro-3,3-dimethyl-butan-2-one（1） and ~1H-[1,2,4]triazol-3-ylamine（2） with equal amount of K_2CO_3 as acid acceptor. The structure of compound 3 was characterized by ~1H NMR, 13 C NMR, HRMS and single-crystal X-ray diffraction. The compound crystallizes in the monoclinic system, space group P21/n with a = 5.7227（8）, b = 27.924（4）, c = 6.2282（7） ？, β = 101.892（11）°, V = 973.9（2） ？~3, Z = 4, T = 180.00（10） K, μ（MoKα） = 0.087 mm^（-1）, Dc = 1.243 g/cm^3, 3832 reflections measured（3.648≤θ≤26.022°）, 1916 unique reflections（Rint = 0.0359, Rsigma = 0.0572） used in all calculations. The final R = 0.0557（I 〉 2σ（I）） and w R = 0.1276（all data）. Bioassay showed that 3 displayed excellent activity as plant growth regulator with inducing lateral root formation and enhancing primary root elongation at 0.27 mmol/L（50 ppm） in soybeen（He Feng-50）. Good water solubility was found with 50 mg in 1 m L of water. Therefore, application of 3 in agriculture is more environmentally friendly due to cosolvent-free condition, and results in improved abiotic-stress tolerance by affecting the root growth. And furthermore, it can be used as a precursor to investigate the function of regulating plant root growth.

circ_0003204 regulates the osteogenic differentiation of human adipose-derived stem cells via miR-370-3p/HDAC4 axis

Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in mesenchymal stem cell osteogenesis.However,how circRNAs regulate hASCs in osteogenesis is still unclear.Herein,we found circ_0003204 was significantly downregulated during osteogenic differentiation of hASCs.Knockdown of circ_0003204 by si RNA or overexpression by lentivirus confirmed circ_0003204 could negatively regulate the osteogenic differentiation of hASCs.We performed dual-luciferase reporting assay and rescue experiments to verify circ_0003204 regulated osteogenic differentiation via sponging miR-370-3p.We predicted and confirmed that miR-370-3p had targets in the 3′-UTR of HDAC4 m RNA.The following rescue experiments indicated that circ_0003204 regulated the osteogenic differentiation of hASCs via miR-370-3p/HDAC4 axis.Subsequent in vivo experiments showed the silencing of circ_0003204 increased the bone formation and promoted the expression of osteogenic-related proteins in a mouse bone defect model,while overexpression of circ_0003204 inhibited bone defect repair.Our findings indicated that circ_0003204 might be a promising target to promote the efficacy of hASCs in repairing bone defects.

A Novel Cellular Autoaggregative Developmentally CRP Regulated Behaviour Generates Massively Chondrule-Like Formations over Surface of Old <i>Escherichia coli</i>K-12 Macrocolony Biofilms

How Escherichia coli bacteria develop a particular colonial, 3-D biofilm morphological pattern is still a poorly understood process. Recently, we reported a new E. coli K-12 morphotype exhibited by old macrocolonies described as volcano-like. The formative developmental process of this morphotype has been presented as a suitable experimental model for the study of 3D patterning in macrocolony biofilms. Here, we report the optical microscopy observations and genetic analysis that have unveiled the existence of a novel autoaggregative behaviour which generates massive lumpiness over the surface of the volcano-like macrocolonies. These lumpy formations are generated by the autoaggregation and strong interaction of tightly packed bacterial cells in structures with a chondrule-like appearance which give the colony’s surface its characteristic microscopic lumpy phenotype. Furthermore, they exhibit different levels of maturation from the edge to the center of the colony. Hence, its generation appears to follow a spatiotemporal program of development during the macrocolony’s morphogenesis. Interestingly, the agar’s hardness influences the morphology exhibited by these formations, with high agar concentration (1.5%, 15 g/L) suppressing its development. This new auto-aggregative E. coli’s behaviour does not require the activity of the biofilm master regulator CsgD, the adhesiveness of flagella, pili type 1, adhesin Ag43, β-1,6-N-acetyl-D-glucosamine polymer-PGA, cellulose or colanic acid, but it is under glucose repression and the control of cAMP receptor protein (CRP). The possible physiological role of these chondrule-like formations in the adaptability of the colony to different stressful environmental conditions is discussed.

骨髓间充质干细胞的体外扩增及其对T淋巴细胞产生IFN-γ和IL-10的影响

为了探讨骨髓间充质干细胞(mesenchymal stem cells,MSC)对T淋巴细胞分泌功能的免疫调节作用,从人骨髓分离培养MSC,通过其形态的均一性及流式细胞术检测表面标志以鉴定其纯度;从外周血分离获得T淋巴细胞,再将MSC分别以不同数量加入到植物血凝素(PHA)刺激的外周血T淋巴细胞和混合淋巴细胞反应(MLR)培养体系及不同浓度MSC培养上清加入混合淋巴细胞反应培养体系共培养后,分别收集上清,ELISA检测IFN-γ和IL-10水平,发现不同细胞数量的MSC对T细胞分泌细胞因子IFN-γ均有抑制,同时促进IL-10分泌,且其抑制和促进均呈剂量依赖性;MSC的不同上清浓度对IFN-γ抑制亦呈浓度依赖性,但未发现对IL-10分泌的影响。表明骨髓MSC在体外可抑制T细胞分泌IFN-γ、促进分泌IL-10,起到免疫调节作用。

Ferroptosis mechanism and Alzheimer's disease

Regulated cell death is a genetically determined form of programmed cell death that commonly occurs during the development of living organisms.This process plays a crucial role in modulating homeostasis and is evolutionarily conserved across a diverse range of living organisms.Ferroptosis is a classic regulatory mode of cell death.Extensive studies of regulatory cell death in Alzheimer’s disease have yielded increasing evidence that fe rroptosis is closely related to the occurrence,development,and prognosis of Alzheimer’s disease.This review summarizes the molecular mechanisms of ferroptosis and recent research advances in the role of ferro ptosis in Alzheimer’s disease.Our findings are expected to serve as a theoretical and experimental foundation for clinical research and targeted therapy for Alzheimer’s disease.

扶正祛毒方对慢性乙肝病毒携带小鼠肝组织γ-干扰素和白介素-10表达影响的研究

目的观察扶正祛毒方对慢性乙肝病毒携带小鼠γ-干扰素( Interferon-γ,IFN-γ)和白介素 10( Interleukin-10,IL-10)表达影响,探讨扶正祛毒方对免疫的调节作用。方法选择雄性 HBV转基因 C57BL/6J 小鼠30 只随机分为模型组、西药组、中药组,同系雄性正常小鼠10 只作为空白组,西药组、中药组分别予以阿德福韦酯药液和扶正祛毒方药液灌胃,模型组、空白组均予以等量生理盐水灌胃。24 周后处死取材,检测血清乙型肝炎核心相关抗原( hepatitis Beantigen,HBeAg)、乙型肝炎表面抗原( hepatitis B surface antigen,HBsAg)水平,丙氨酸氨基转移酶( alanine aminotransferase,ALT)、谷草转氨酶( aspartate transaminase,AST)含量,尾尖乙型肝炎病毒脱氧核糖核酸( hepatitis Bvirus deoxyribonucleic acid,HBV DNA)相对表达量,肝组织γ-干扰素、白介素-10 含量。结果( 1)ALT、AST:模型组较空白组显著升高( P < 0. 05),西药组和中药组较模型组显著降低( P < 0. 05)。( 2) HBeAg、HBsAg:模型组 HBeAg、HBsAg 阳性率均为 100%,空白组均为 0%,西药组、中药组HBeAg 阴转率分别为 28. 6%、42. 9%。( 3) HBV DNA:模型组较空白组显著升高( P <0. 05),中药组较模型组显著降低( P <0. 05)。( 4)γ-干扰素:模型组较空白组显著降低( P <0. 05),中药组显著高于模型组( P <0. 05)。( 5)白介素-10:模型组较空白组显著升高( P <0. 05),中药组显著低于模型组和西药组( P <0. 05)。结论( 1)用药 24 周后,扶正祛毒方对慢乙肝病毒携带小鼠肝脏炎症反应无加剧,并能降低血清转氨酶水平;( 2)用药 24 周后,扶正祛毒方对慢性乙肝病毒携带小鼠病毒复制有抑制作用;( 3)用药 24 周后,扶正祛毒方可以提高慢性乙肝病毒携带小鼠肝组织γ-干扰素表达,并抑制白介素-10 表达,这可能是其抑制病毒复制的重要机制之一。

郁证发微(四十八)--郁证斑秃论

论文探讨郁证性斑秃的病脉症治。郁证性斑秃与情志因素致病密切相关,其病机多现肝气郁结和(或)心脾两亏。中医肝脏象情绪量表分析、中医体质研究均发现斑秃患者具有七情不遂、肝气郁结的特点。临床治疗除了养血填精生发以外,毋忘疏肝解郁、养心安神等从郁论治或辅助从郁论治。现代医学"神经-内分泌-免疫调节"理论也证实精神因素与斑秃具有密切的关系。

Protein-spatiotemporal partition releasing gradient porous scaffolds and anti-inflammatory and antioxidant regulation remodel tissue engineered anisotropic meniscus

Meniscus is a wedge-shaped fibrocartilaginous tissue,playing important roles in maintaining joint stability and function.Meniscus injuries are difficult to heal and frequently progress into structural breakdown,which then leads to osteoarthritis.Regeneration of heterogeneous tissue engineering meniscus(TEM)continues to be a scientific and translational challenge.The morphology,tissue architecture,mechanical strength,and functional applications of the cultivated TEMs have not been able to meet clinical needs,which may due to the negligent attention on the importance of microenvironment in vitro and in vivo.Herein,we combined the 3D(three-dimensional)-printed gradient porous scaffolds,spatiotemporal partition release of growth factors,and anti-inflammatory and anti-oxidant microenvironment regulation of Ac2-26 peptide to prepare a versatile meniscus composite scaffold with heterogeneous bionic structures,excellent biomechanical properties and anti-inflammatory and anti-oxidant effects.By observing the results of cell activity and differentiation,and biomechanics under anti-inflammatory and anti-oxidant microenvironments in vitro,we explored the effects of anti-inflammatory and anti-oxidant microenvironments on construction of regional and functional heterogeneous TEM via the growth process regulation,with a view to cultivating a high-quality of TEM from bench to bedside.

lncRNACNN3-206 activates intestinal epithelial cell apoptosis and invasion by sponging miR-212, an implication for Crohn’s disease

BACKGROUND Statistics indicate that the incidence of Crohn’s disease(CD)is rising in many countries.The poor understanding on the pathological mechanism has limited the development of effective therapy against this disease.Previous studies showed that long noncoding RNAs(lncRNAs)could be involved in autoimmune diseases including CD,but the detailed molecular mechanisms remain unclear.AIM To identify the differentially expressed lncRNAs in the intestinal mucosa associated with CD,and to characterize their pathogenic role(s)and related mechanisms.METHODS The differential expression of lncRNAs was screened by high-throughput RNA sequencing,and the top candidate genes were validated in an expanded cohort by real-time PCR.The regulatory network was predicted by bioinformatic software and competitive endogenous RNA analysis,and was characterized in Caco-2 and HT-29 cell culture using methods of cell transfection,real-time PCR,Western blotting analysis,flow cytometry,and cell migration and invasion assays.Finally,these findings were confirmed in vivo using a CD animal model.RESULTS The 3'end of lncRNACNN3-206 and the 3’UTR of Caspase10 contain highaffinity miR212 binding sites.lncRNACNN3-206 expression was found to be significantly increased in intestinal lesions of CD patients.Activation of the lncRNACNN3-206-miR-212-Caspase10 regulatory network led to increased apoptosis,migration and invasion in intestinal epithelial cells.Knockdown of lncRNACNN3-206 expression alleviated intestinal mucosal inflammation and tissue damage in the CD mouse model.CONCLUSION lncRNACNN3-206 may play a key role in CD pathogenesis.lncRNACNN3-206 could be a therapeutic target for CD treatment.

异位高表达OCT4的人骨髓间充质干细胞对T淋巴细胞功能的影响

目的:探讨异位高表达OCT4的人骨髓间充质干细胞(MSC)在体外对T细胞增殖、活化及分泌功能的影响。方法:分离健康儿童外周血单个核细胞,使用Anti-CD3/CD28单克隆抗体体外激活T淋巴细胞,白介素(IL)2体外刺激培养的T细胞1周。建立异位高表达OCT4的MSC(MSC-OCT4)与活化T细胞的体外共培养体系,收集共培养1周后的上清液,流式细胞仪测定Th1/Th2细胞因子(IL-2、IL-4、IL-6、IL-10、肿瘤坏死因子-α、γ干扰素)水平。收集共培养1周后的淋巴细胞,通过Countstar软件进行计数,用流式细胞仪测定T细胞及活化T细胞亚群的比例后,计算出绝对值,以均数±标准差表示。结果:与对照单独T细胞培养组相比,MSC及MSC-OCT4均能显著抑制CD3^(+)T细胞、CD3^(+)CD4^(+)T细胞和CD3^(+)CD8^(+)T细胞增殖。与MSC相比,MSC-OCT4能更好地抑制CD3^(+)CD8^(+)T细胞增殖(P=0.049),且主要抑制早期T细胞活化。与对照单独T细胞培养组相比,MSC及MSC-OCT4均能显著下调细胞因子IL-2和γ干扰素水平。与T细胞共培养1周后,MSC与MSC-OCT4组细胞因子IL-6水平显著增加。与对照MSC组相比,MSC-OCT4组在共培养1周后活细胞数更高(P=0.019),且能耐受更高浓度丝裂霉素C的抑制增殖作用。结论:MSC及MSC-OCT4在体外均能抑制IL-2刺激的T细胞增殖及活化,过表达OCT4后MSC能更强地抑制CD3^(+)CD8^(+)T细胞增殖,且具有更好的体外增殖能力,可能具有更好更持久地调节Th1/Th2造血因子平衡的作用。

Up-regulation of LPS-induced iNOS activity in dibutyryl cyclic AMP-differentiated rat astrocytes

目的：研究ｄＢｃＡＭＰ诱导分化成熟后的星状胶质细胞内脂多糖诱导型一氧化氮合酶活力的诱导表达情况．方法：采用荧光分光光度法和胍胺酸生成实验考察星状胶质细胞内一氧化氮合酶的活力，并通过免疫细胞化学和光学显微镜考察细胞的分化情况．结果：经ｄＢｃＡＭＰ诱导后，星状胶质细胞分化成星型细胞，ＧＦＡＰ表达增强．ｄＢｃＡＭＰ显著增强ＬＰＳ诱导产生的亚硝酸盐水平的提高；ｄＢｃＡＭＰ／ＬＰＳ共同诱导２４ｈ后，使ＮＯＳ活力比ＬＰＳ单独诱导提高４倍．ＬＮＡＭＥ，环己亚胺或放线菌素Ｄ都可分别抑制ｄＢｃＡＭＰ／ＬＰＳ的共同诱导作用．结论：ｄＢｃＡＭＰ可以上调ＬＰＳ对星状胶质细胞内ｉＮＯＳ的诱导作用．

Decreased miR-325-5p Contributes to Visceral Hypersensitivity Through Post-transcriptional Upregulation of CCL2 in Rat Dorsal Root Ganglia

Chronic visceral hypersensitivity is an important type of chronic pain with unknown etiology and pathophysiology. Recent studies have shown that epigenetic regulation plays an important role in the development of chronic pain conditions. However, the role of mi RNA-325-5 p in chronic visceral pain remains unknown. The present study was designed to determine the roles and mechanism of mi RNA-325-5 p in a rat model of chronic visceral pain.This model was induced by neonatal colonic inflammation(NCI). In adulthood, NCI led to a significant reduction in the expression of mi RNA-325-5 p in colon-related dorsal root ganglia(DRGs), starting to decrease at the age of4 weeks and being maintained to 8 weeks. Intrathecal administration of mi RNA-325-5 p agomir significantly enhanced the colorectal distention(CRD) threshold in a time-dependent manner. NCI also markedly increased the expression of CCL2(C-C motif chemokine ligand 2) in colon-related DRGs at the m RNA and protein levels relative to age-matched control rats. The expression of CXCL12, IL33, SFRS7, and LGI1 was not significantlyaltered in NCI rats. CCL2 was co-expressed in Neu Npositive DRG neurons but not in glutamine synthetasepositive glial cells. Furthermore, CCL2 was mainly expressed in isolectin B4-binding-and calcitonin generelated peptide-positive DRG neurons but in few NF-200-positive cells. More importantly, CCL2 was expressed in mi R-325-5 p-positive DRG neurons. Intrathecal injection of mi RNA-325-5 p agomir remarkably reduced the upregulation of CCL2 in NCI rats. Administration of Bindarit, an inhibitor of CCL2, markedly raised the CRD threshold in NCI rats in a dose-and time-dependent manner. These data suggest that NCI suppresses mi RNA-325-5 p expression and enhances CCL2 expression, thus contributing to visceral hypersensitivity in adult rats.

Post-translational inhibitory regulation of acid invertase induced by fructose and glucose in developing apple fruit

Acid invertase (EC 3.2.1.26) is one of the key enzymes involved in the carbohydrate sink-organ development and the sink strength modulation in crops. The experiment conducted with 'Starkrimson' apple (Malus domestica Borkh) fruit showed that, during the fruit development, the activity of acid invertase gradually declined concomitantly with the progressive accumulation of fructose, glucose and sucrose, while Western blotting assay of acid invertase detected a 30 ku peptide of which the immuno-signal intensity increased during the fruit development. The immuno-localization via immunogold electron microscopy showed that, on the one hand, acid invertase was mainly located on the flesh cell wall with numbers of the immunosignals present in the vacuole at the late stage of fruit development; and on the other hand, the amount of acid invertase increased during fruit development, which was consistent with the results of Western blotting. The in vivo pre-incubation of fruit discs with soluble sugars showed that the activity of extractible acid invertase was inhibited by fructose or glucose, while Western blotting did not detect any changes in apparent quantity of the enzyme nor other peptides than 30 ku one. So it is considered that fructose and glucose induced the post-translational or translocational inhibitory regulation of acid invertase in developing apple fruit. The mechanism of the post-translational inhibition was shown different from both the two previously reported ones that proposed either the inhibition by hexose products in the in vitro chemical reaction equilibrium system or the inhibition by the proteinaceous inhibitors. It was hypothesized that fructose and glucose might induce acid invertase inhibition by modulating the expression of some inhibition-related genes or some structural modification of acid invertase.

Mast Cell-Derived Exosomes at the Stimulated Acupoints Activating the Neuro-immune Regulation

Exosomes are cell-derived vesicles that take part in intercellular signaling. Research has shown that acupuncture is closely related to affecting the functions of the mast cells in the local region of the acupoint, and stimulating the afferent nerve. Mast cells have a connection with the conduction within the meridians, and play an important role in immuno-regulation. The ＇synapse-like＇ connection between the mast cells and nerve endings is the basis for the exchange of information between these two tissues. Exosome mediates mast exchange of information between mast cells and the nerves, starting the process of neuro-immuno regulation. Therefore, we propose that mast cell-derived exosomes mediate the neuro-immuno regulation at the local site of acupuncture, and this is one of the key factors resulting in the effectiveness of acupuncture.