Multi-genome evolutionary study of the ABC1 gene family and identification of the pleiotropic effects of OsABC1-13 in rice development

In four rice genomes,85 ABC1-family genes were identified by comparative genomics,evolution,genetics,and physiology.One,OsABC1-13,was shown by knockdown and knockout experiments to affect plant height,grain size,and photosynthetic capability.

乳腺癌患者病理特征与Bcl-2、CXCL13、PAX8表达情况的关系分析

目的分析乳腺癌患者病理特征与B细胞淋巴瘤/白血病-2基因(Bcl-2)、趋化因子配体13(CXCL13)、配对盒基因8抗体(PAX8)表达情况的关系。方法收集2021年1月至2023年1月该院收治的160例乳腺癌患者临床资料。采用免疫组化法对其癌组织与癌旁组织Bcl-2、CXCL13、PAX8表达情况进行检测,并分析3项指标与患者病理特征的关系。结果与癌旁组织比较,癌组织Bcl-2、CXCL13、PAX8阳性率更高,差异有统计学意义(P<0.05)。与雌激素受体(ER)阴性、肿瘤最大径≥3 cm、孕激素受体(PR)阴性患者比较,ER阳性、肿瘤最大径<3 cm、PR阳性患者中Bcl-2高表达占比更高,差异有统计学意义(P<0.05);与无淋巴结转移、Ⅰ~Ⅱ期患者比较,淋巴结转移、Ⅲ~Ⅳ期患者中CXCL13高表达占比更高,差异有统计学意义(P<0.05);与Ⅰ~Ⅱ期、高/中分化、无淋巴结转移患者比较,Ⅲ~Ⅳ期、低分化、有淋巴结转移患者中PAX8高表达占比更高,差异有统计学意义(P<0.05)。ER、PR表达情况与Bcl-2表达情况呈正相关(P<0.05),肿瘤最大径与Bcl-2表达情况呈负相关(P<0.05);临床分期、淋巴结转移情况与CXCL13、PAX8表达情况呈正相关(P<0.05);分化程度与PAX8表达情况呈负相关(P<0.05)。结论乳腺癌患者Bcl-2、CXCL13、PAX8表达情况对疾病的发生和发展具有明显影响,有望成为评估乳腺癌患者病情严重程度的标志物。

Association between the interleukin-13 gene and development of chronic obstructive pulmonary disease in southern Chinese Han population： a case-control study

Background Interleukin-13 （IL-13） has been implicated to be responsible for recruitment of inflammatory cells from the blood to the lung,regulation of matrix metalloproteinase and induction of mucin production and secretion in chronic obstructive pulmonary disease （COPD）.We determined plasma IL-13 levels in patients with COPD and investigated its association with common polymorphisms of IL-13 gene in a case-control study.Methods We genotyped 160 cases and 175 control subjects in a local hospital using Mass-ArrayTM Technology Platform then tested the association of four SNPs in IL-13 （rs1295685,rs1800925,rs1881457,rs20541） with COPD,and then determined plasma IL-13 levels in patients with COPD and controls.Results Association was found between IL-13 gene SNPs （rs20541 and rs1800925） and an increased risk of COPD.By linkage disequilibrium （LD） analysis,two blocks （rs1881457 and rs1800925; rs20541 and rs1295685） were found.The risk of COPD was found associated with the IL-13 gene polymorphism among southern Chinese Han population.Plasma IL-13 level was increased in COPD patients compared with controls.Conclusions The polymorphism of the IL-13 gene is associated with an increased risk of COPD in southern Chinese Han population.Plasma IL-13 levels were found elevated in patients with COPD.

Inhibition of interleukin-13 gene expression in T cells through GATA-3 pathway by arsenic trioxide

Aarsenic trioxide （AT） has a long history of use in both traditional Chinese medicine and in modem medicine in asthma therapy. Recently, Yin et al＇ found that AT even at small doses reduced the airway inflammation of sensitized guinea pigs. However the mechanism underlying this is still largely unknown. Interleukin 13 （IL-13）, as one of the important TH2 cytokines, plays an important role in asthma pathogenesis through promoting eosinophilic inflammation,

6-Gingerol, asarinin, and deoxyschizandrin improve bronchial epithelium functions in an interleukin-13einduced BEAS-2B cell model

Objective:To explore the effects of 6-gingerol,asarinin,and deoxyschizandrindthe main components of Zingiber officinale(Willd.)Rosc.(Gan Jiang),Asarum heterotropoides f.var.mandshuricum(Maxim.)(Xi Xin),and Schisandra chinensis(Turcz.)Baill.(Wu Wei Zi),respectivelydon an interleukin(IL)-13einduced BEAS-2B cell model in vitro.Methods:The BEAS-2B cell model was established using 25 ng/mL IL-13 combined with 1%fetal bovine serum(FBS)in vitro.Mitoquinone mesylate(Mito-Q)treatment was used as a positive control group,and different concentrations of 6-gingerol,asarinin,and deoxyschizandrin were used to treat the models.The level of reactive oxygen species(ROS)production was detected by flow cytometry.The expression levels of LC3B,Beclin1,adenosine 50-monophosphate(AMP)eactivated protein kinase(AMPK),phosphory-lated-AMPeactivated protein kinase(P-AMPK),dynamin-related protein 1(DRP1),and mitochondrial fusion protein 2(MFN2)were detected by Western blot.Mitochondrial membrane potential(MMP)assay kit with JC-1 was utilized to detect the level of MMP.Results:The BEAS-2B cells exposed to 25 ng/mL IL-13 with 1%FBS showed an increased ROS level and a decreased MMP.6-Gingerol,asarinin,and deoxyschizandrin were able to downregulate ROS level and upregulate the MMP in the BEAS-2B model.Asarinin and deoxyschizandrin reduced the expression of autophagy protein LC3B,while deoxyschizandrin significantly increased the expression of DRP1 in the BEAS-2B model.Conclusion:6-Gingerol,asarinin,and deoxyschizandrin can reduce ROS generation and increase MMP,but have different regulatory effects on the expression of autophagy protein and mitochondrial mitotic protein.The three components have both synergistic and complementary effects in classic medicine compatibility.This study may provide an innovative strategy to reduce the lung inflammation related to IL-13.

自噬蛋白(ATG13)激活IFNα表达的机制研究

自噬相关蛋白基因13(Autophagy-related gene 13,atg13)在细胞自噬的发生过程中发挥着关键作用。然而,其在激活干扰素α(Interferon-α,IFN-α)的过程中所发挥的作用尚未阐明。基于对atg13基因的物种特异性的遗传特征,本研究选取人源atg13基因及人源胚胎肾细胞系(human embryonic kidney cell line,HEK 293T),作为研究atg13基因过表达与目标细胞中激活IFN-α信号通路的试验平台。结果表明atg13基因在刺激IFN-α产生的能力方面明显强于其激活IFN-β的活性。虽然atg13基因与IFN信号通路中的不同信号分子(MyD88,IKKα,TRAF6和IRF7)在转录及翻译水平上均有不同程度的相关性,但atg13基因过表达从而激活IRF7的磷酸化是IFN-α产生的关键因素。此外,ATG13蛋白与干扰素刺激应答原件(Interferon-stimulated response element,ISRE)的强烈互作也是atg13基因过表达激活IFN-α产生的重要环节。

基质金属蛋白酶13在靶向治疗骨关节炎的研究进展

骨关节炎(osteoarthritis,OA)是一类常见的慢性退行性疾病,可导致关节软骨、滑膜、软骨下骨等多组织损伤。目前,该疾病的发病机制尚未完全阐明。研究发现基质金属蛋白酶(matrix metalloproteinase,MMPs)与该疾病的发病过程相关,特别是MMP13,其在机体内的表达水平与OA关系密切。近年来,针对MMP13在该疾病发生发展中的作用,靶向MMP13治疗OA具有诸多进展,选择性MMP13抑制剂较广谱MMPs抑制剂具有选择性强、毒副作用低、效能高等优点,但需要频繁给药以维持药物在体内的有效浓度;将与MMP13 mRNA具有选择性互补的小干扰RNA注射入关节腔可有效降低体内MMP13蛋白产生并抑制多种炎症因子表达,且单次注射可较长时间维持药物效果;利用CRISPR⁃Cas9基因编辑技术靶向消融MMP13基因以减弱软骨细胞外基质蛋白的降解从而治疗OA的方式在动物体内外模型中均具有较好效果,但该技术在人体内长期应用的安全性也引发人们的思考。靶向MMP13治疗OA是医学界及科研领域的重要研究方向,未来结合更前沿的生物医学技术将为探索治疗OA提供新的策略。

Interleukin-35: A key player managing pre-diabetes and chronic inflammatory type 1 autoimmune diabetes

Interleukin-35(IL-35)is a novel protein comprising IL-12αand IL-27βchains.The IL12A and EBI3 genes are responsible for its production.The study of IL-35 has experienced a substantial increase in interest in recent years,as demonstrated by many research papers.Recent clinical studies have shown that individuals who do not have a C-peptide have notably reduced amounts of IL-35 in their blood serum.This is accompanied by a drop in the percentage of IL-35+Treg cells,regulatory B cells,and CD8+FOXP3+cells that produce IL-35.This article em-phasizes the potential significance of IL-35 expression in governing the immune response and its involvement in chronic inflammatory autoimmune diabetes in pancreatic inflammation.It demonstrates IL-35's ability to regulate cytokine proportions,modulate B cells,and protect against autoimmune diabetes.However,further investigation is necessary to ascertain the precise mechanism of IL-35,and meticulous planning is essential for clinical studies.

Interleukin-13 and age-related macular degeneration

AIM：To identify the effects of interleukin（IL）-13 on retinal pigment epithelial（RPE） cells and the IL-13 level in aqueous humor of age-related macular degeneration（AMD） patients.METHODS：IL-13 levels in aqueous humor specimens from AMD patients were detected with enzyme-linked immunosorbent assay（ELISA）.ARPE-19 cells were treated with 10 ng/m L IL-13 for 12,24,and 48 h.The cell proliferaton was evaluated by the MTS method.The m RNA and protein levels of α-SMA and ZO-1 were evaluated with quantitative real-time polymerase chain reaction（q RT-PCR） and Western blot respectively.The expression of tumor necrosis factor-α（TNF-α）,transforming growth factor-β（TGF-β） and vascular endothelial growth factor（VEGF） were assessed by ELISA.RESU LTS：IL-13 levels in the aqueous humor of patients with AMD were significantly higher than those in the control（167.33±17.64 vs 27.12±5.65 pg/m L;P〈0.01）.In vit ro,IL-13 of high concentrations（10,15,and 20 ng/m L） inhibited ARPE-19 cell proliferation.α-SMA m RNA in ARPE-19 cell were increased（1.017±0.112 vs 1.476±0.168;P〈0.001） and ZO-1 decreased（1.051±0.136 vs 0.702±0.069;P〈0.001） after treated with 10 ng/m L IL-13 for 48 h.The protein expression of α-SMA and ZO-1 also showed the same tendency（α-SMA：P=0.038;ZO-1：P=0.008）.IL-13 significantly reduced the level of TNF-α（44.70±1.67 vs 31.79±3.53 pg/m L;P=0.005） at 48 h,but the le vel of TGF-β2 was significantly increased from 34.44±2.92 to 57.61±6.31 pg/m L at 24h（P=0.004） and from 61.26±1.11 to 86.91±3.59 pg/m L at 48h（P〈0.001）.While expressions of VEGF didn＇t change after IL-13 treatment.CONCLUSION：IL-13 in vitr o inhibit ARPE-19 cell proliferation and expression in the aqueous may be associated with AMD.

沉默PRR13基因对结直肠癌细胞增殖和凋亡的影响及其可能机制

目的探讨沉默富脯氨酸多肽13(PRR13)基因对结直肠癌细胞增殖、凋亡的影响,并基于自噬相关蛋白、免疫细胞相关因子和Wnt/β⁃连环蛋白信号通路探讨其可能的作用机制。方法(1)检测正常人结直肠上皮细胞(FHC细胞)及人结直肠癌细胞(HCT116细胞、HT⁃29细胞、LS174T细胞、SW480细胞、SW620细胞、LoVo细胞和HR⁃8348细胞)的PRR13 mRNA表达水平,选择表达水平最高的人结直肠癌细胞进行后续实验。(2)将HR⁃8348细胞分为对照组、空载体组、PRR13⁃小干扰RNA(siRNA)组、IWR⁃1组、PRR13⁃siRNA+IWR⁃1组。不给予对照组任何干预,给予空载体组转染阴性对照siRNA,给予PRR13⁃siRNA组转染PRR13⁃siRNA,给予IWR⁃1组Wnt/β⁃连环蛋白信号通路抑制剂IWR⁃1干预,给予PRR13⁃siRNA+IWR⁃1组转染PRR13⁃siRNA并给予IWR⁃1干预。检测各组的PRR13 mRNA表达水平、细胞存活率、细胞凋亡率、γ⁃干扰素水平、白细胞介素(IL)⁃4水平,以及微管相关蛋白1轻链3Ⅱ(MAP1LC3Ⅱ)、Beclin 1蛋白、β⁃连环蛋白、糖原合成酶激酶3β(GSK⁃3β)蛋白、Wnt11蛋白的表达水平。结果(1)除SW620细胞外,其他6种人结直肠癌细胞的PRR13 mRNA表达水平均高于人结直肠上皮FHC细胞,且HR⁃8348细胞中的PRR13 mRNA表达水平高于其他人结直肠癌细胞(P<0.05)。(2)与对照组及空载体组相比,其他3组的PRR13 mRNA表达水平、细胞存活率、IL⁃4水平降低,MAP1LC3Ⅱ蛋白、Beclin 1蛋白、β⁃连环蛋白、GSK⁃3β蛋白、Wnt11蛋白表达水平下调,细胞凋亡率及γ⁃干扰素表达水平升高(P<0.05)。与PRR13⁃siRNA组及IWR⁃1组相比,PRR13 siRNA+IWR⁃1组上述指标的改变更为明显(P<0.05),但PRR13⁃siRNA组及IWR⁃1组的上述指标差异并无统计学意义(P>0.05)。结论沉默PRR13基因可抑制结直肠癌细胞活性,促进其凋亡,其机制可能与调控Th分化、抑制自噬相关蛋白活性及Wnt/β⁃连环蛋白信号通路激活有关。联合沉默PRR13基因与抑制Wnt/β⁃连环蛋白信号通路,或许可更好地发挥抗结直肠癌的作用。

miRNAs-MUC 13调控体系中与仔猪腹泻有关的基因与通路

为研究microRNAs-Mucin 13(miRNAs-MUC 13)调控体系中与仔猪腹泻相关的分子调控机制,对miRNAs-MUC 13调控体系中与仔猪腹泻有关的基因与通路进行了发掘与验证。用NCBI的blastn工具比对了小鼠MUC 13基因和猪基因库中的核酸序列,发现两者的相似性在74%~87%,同源性较高。以小鼠为模型,选择miRDB、mirTarBase、TargetScan数据库预测调控MUC 13基因的miRNA序列,通过StarBase数据库预测这些miRNA序列的靶基因,得到36个靶基因交集。用Cytoscape软件围绕MUC 13和这36个靶基因构建仔猪腹泻基因互作网络,该网络包含19个筛选到的靶基因,基于MCOMD算法分析获得4个分子复合物集团。通过DAVID平台对这4个分子复合物进行基因和通路富集分析,结果发现,靶基因SRF和STAG 2及其参与的有关通路与仔猪腹泻相关。以大肠埃希菌200和仔猪肠上皮细胞系(IPEC-1)作为试验材料,通过体外细胞试验设计仔猪腹泻模型进行验证,实时荧光定量PCR(qRT-PCR)检测结果显示,试验组MUC 13、SRF、IL-6、MAPK、STAG 2、NF-κB、TLR 7等基因的表达量均显著(P<0.05)高于对照组,由此推测,SRF与STAG 2基因在miRNAs-MUC 13调控体系中分别通过MAPK、IL6、Toll-like受体信号通路参与仔猪腹泻调控。围绕miRNAs-MUC 13调控体系发掘了4个分子复合物集团和2个核心基因(SRF和STAG 2)并进行了试验验证,这为靶向治疗仔猪腹泻提供了理论参考和科学依据。

Association of polymorphisms of interleukin-18 gene promoter region with chronic hepatitis B in Chinese Han population

AIM: To investigate the polymorphisms of interleukin-18 (IL-18) gene promoters, and to disclose whether such polymorphisms are associated with susceptibility to chronic hepatitis B in Chinese Han population. METHODS: Using polymerase chain reaction with sequence specific primers (PCR-SSP) method, the single nucleotide polymorphisms (SNPs) of the promoter region of IL-18 gene at position -607 and -137 were detected in 231 patients with chronic hepatitis B and 300 normal controls. RESULTS: Allele C at position -607 in the promoter of IL-18 gene was detected in 48.7% of normal controls and 51.9% of patients, while allele A at position -607 was detected in 51.3% of normal controls and 48.1% of patients. The frequencies of -607CC, -607 CA and -607AA genotypes in normal controls were 22.0%, 53.3% and 24.7% respectively and in chronic hepatitis B patients were 26.8%, 50.2% and 23.0% respectively. Allele G at position -137 in the promoter of IL-18 gene was detected in 82.3% of normal controls and 88.5% of chronic hepatitis B patients, while allele C at position -137 was detected in 17.7% of normal controls and 11.5% of patients. The frequencies of -137GG, GC and CC genotype were 67.3%, 30.0% and 2.7% in normal controls respectively, while in chronic hepatitis B patients were 78.8%, 19.5% and 1.7% respectively. The frequency of-137GG genotype in chronic hepatitis B groups was significantly higher than that in normal controls (x2=8.55, P=0.003 <0.05), whereas the frequencies of -607C/-137C and -607A/-137C haplotypes in chronic hepatitis B groups were significantly lower than that in normal controls. The association between genotypes of IL-18 promoter region polymorphisms and HBV copies showed that the frequency of -607AA genotype in high HBV-DNA copies groups was lower than that in low HBV-DNA copies groups (x2=6.03, P=0.014 <0.05). CONCLUSION: The polymorphisms of the promoter region of IL-18 gene at position -607 and -137 are closely associated with susceptibility to chronic hepatitis B. The people with allele C at position -137 in the promoter of IL-18 gene may be protected against HBV infection; moreover AA genotype at position -607 may be closely linked to inhibit HBV-DNA replication. These findings give some new clues to the study of pathogenesis of chronic hepatitis B.

Interleukin-1β gene polymorphism associated with hepatocellular carcinoma in hepatitis B virus infection

AIM： To examine the effect of interleukin-l-beta （IL-113） promoter region C-511T and IL-1 receptor antagonist （IL-1RN） polymorphism among the patients with chronic hepatitis B virus （HBV） infection （HCC and non-HCC）. METHODS： Genomic DNA from 136 Thai patients with chronic HBV infection （HCC =46 and non-HCC= 90） and 152 healthy individuals was genotyped for IL-113 gene polymorphism （-511） using polymerase chain reaction with sequence specific primers （PCR-SSP）. The variable number of tandem repeats （VNTR） of IL-1RN gene was assessed by a PCR-based assay. The association between these genes and status of the disease was evaluated by X^2 test. RESULTS： IL-1B-511 genotype c/c was found to be significantly different in patients with HCC when compared with healthy individuals （P = 0.036, OR = 2.29, 95%CI = 1.05-4.97） and patients without HCC （P=0.036, OR= 2.52, 95%CI=1.05-6.04）. Analysis of allele frequencies of IL-1B-511 showed that IL-1B-511 C allele was also significantly increased in patients with HCC, compared to that in healthy control （P=0.033, OR= 1.72, 95%CI=1.04-2.84）. However, no significant association in IL-1RN gene was found between the two groups. CONCLUSION： IL-1B-511C allele, which may be associated with high IL-1B production in the liver, is a genetic marker for the development of HCC in chronic hepatitis B patients in Thai population.

Downregulation of SL-ZH13 transcription factor gene expression decreases drought tolerance of tomato

Zinc finger-homeodomain proteins(ZF-HDs) are transcription factors that regulate plant growth,development,and abiotic stress tolerance.The SL-ZH13 gene was found to be significantly upregulated under drought stress treatment in tomato(Solanum lycopersicum) leaves in our previous study.In this study,to further understand the role that the SL-ZH13 gene plays in the response of tomato plants to drought stress,the virus-induced gene silencing(VIGS) method was applied to downregulate SL-ZH13 expression in tomato plants,and these plants were treated with drought stress to analyze the changes in drought tolerance.The SL-ZH13 silencing efficiency was confirmed by quantitative real-time PCR(qRT-PCR) analysis.In SL-ZH13-silenced plants,the stems wilted faster,leaf shrinkage was more severe than in control plants under the same drought stress treatment conditions,and the mean stem bending angle of SL-ZH13-silenced plants was smaller than that of control plants.Physiological analyses showed that the activity of superoxide dismutase(SOD) and peroxidase(POD) and the content of proline(Pro) in SL-ZH13-silenced plants were lower than those in control plants after 1.5 and 3 h of drought stress treatment.The malondialdehyde(MDA) content in SL-ZH13-silenced plants was higher than that in control plants after 1.5 and 3 h of drought stress treatment,and H2O2 and O2^-· accumulated much more in the leaves of SL-ZH13-silenced plants than in the leaves of control plants.These results suggested that silencing the SL-ZH13 gene affected the response of tomato plants to drought stress and decreased the drought tolerance of tomato plants.

Association Between IL-13 Gene Polymorphism and Asthma in Han Nationality in Hubei Chinese Population

Summary: In order to investigate whether Arg110Gln polymorphism in the coding region of the IL-13 gene is associated with asthma and total plasma IgE level in Han nationality in Hubei Chinese population, the allele frequency of 4257(g/a) site and Arg110Gln genotype of IL-13 was detected by using restriction fragment length polymorphism in Han nationality in Hubei Chinese population including 43 asthmatic children, 45 asthmatic adults, 31 control children and 46 control adults. Total plasma IgE was measured by Chemiluminescence assay. The results showed that the frequency of allele A at 4257 bp of IL-13 in children and adults was 0.39 and 0.32, respectively. The GlnGln form of Arg110Gln polymorphism of IL-13 gene was associated with susceptibility of asthma and elevated total plasma IgE in children (P =0.030 and 0.0009, respectively), but not with them in adults (P=0.219 and 0.174, respectively). Our results suggest that the Arg110Gln polymorphism of IL-13 gene is associated with susceptibility of asthma and elevated total plasma IgE in Chinese children of Han nationality in Hubei, but not with them in adults.

Construction of eukaryotic expression vector of human S100A13 gene and its effect on proliferation of human thyroid cancer cell line TT

Objective To investigate the effect of exogenous S100A13 gene overexpression on the proliferation of human thyroid cancer cell line TT.Methods The recombinant ORF of S100A13 tagged with six histidines at the 5' end was subcloned into the pcDNA3.2/V5/GW/D-TOPO vector and sequenced.The eukaryotic expression plasmid pcDNA3.2/V5 /GW/D-S100A13 and empty vector pcDNA3.2/V5/GW/D were transfected into TT cells.The positive clones were selected by G418.The expressions of S100A13 mRNA and protein were detected by real time reverse transcription-polymerase chain reaction(RT-PCR) and Western blot.The effect of S100A13 on cell proliferation and cell cycle was evaluated by cell growth curve,MTT colorimetric assay and flow cytometry.Results S100A13 gene tagged with six histidines at the 5 ' end was confirmed to be inserted into the pcDNA3.2/V5/GW/D vector correctly.TT-S100A13-V5 cells,which over-expressed S100A13,were constructed successfully.TT-S100A13-V5 cells grew much faster than TT-V5 and TT cells(P <0.001).The proportions of both S and G2/M phase cells were significantly higher in TT-S100A13-V5 cells than those in TT-V5 and TT cells(P <0.001).Conclusion The eukaryotic expression vector containing human S100A13 gene has been successfully constructed,which highly expresses S100A13 in TT cells.Exogenous S100A13 gene overexpression accelerates TT cell proliferation and drives the cell cycle progression of TT cells from G0/G1 phase to S and G2/M phases.

Interleukin-13 inhibits cytokines synthesis by blocking nuclear factor-κB and c-Jun N-terminal kinase in human mesangial cells

Objective： Monocytes/macrophages, proinflammatory cytokines and chemokines are important in the pathogenesis of glomerulonephritis. Interleukin （IL） -13 has been shown to exert potent anti-inflammatory properties. This study was designed to investigate the effect of IL-13 on the expression of proinflammatory cytokines, chemokines and profibrogenic cytokines and the involved molecular mechanism in cultured human mesangial cells （HMCs）. Methods： The expressions of proinflammatory cytokines, chemokines and profibrogenic cytokines were determined by ribonuclease protection assay （RPA）. Activity of nuclear factor-kappa B （NF-κB） and activa- tor protein-1 （AP-1） was examined by electrophoretic mobility shift assay （EMSA）. NF-κB subunit p65 nuclear transportation and c-Jun N-terminal kinase （JNK） activity were assayed by immunoblot. Results： Recombinant IL-13 inhibited tumor necrosis factor-α （TNF-u）, IL-1α, IL-1β, monocyte chemoattractant protein-1 （MCP-1）, IL-8, and transforming growth factor-β1 （TGF-β1） mRNA expressions in a dose-dependent manner. Lipopoly- sacchorides （LPS） dramatically increased NF-κB DNA binding activity of HMCs, which was inhibited by IL-13 in a dose-dependent manner. LPS-activated NF-κB contained p50 and p65 dimers, but not c-Rel subunit. IL-13 blocked LPS-induced NF-κB subunit p65. LPS stimulated JNK/AP-1 activation, which was inhibited by IL-13 in a dose-dependent manner. Conclusion： IL-13 inhibits proinflammatory cytokines, chemokines, and profibrogenic cytokines synthesis by blocking NF-κB and JNK/AP-1 activation. These observations point to the importance of IL-13 in the modulation of inflammatory processes in the renal glomerulus.

Interleukin-8 gene polymorphism is associated with acute coronary syndrome in a Han Chinese population

Background Acute coronary syndrome(ACS) is one of the most common forms of heart diseases.Recent studies have revealed that interleukin(IL)-8 plays a kev role in the development of atherosclerosis plaque and its complications, but the relationship of its common variants with ACS has not been extensively studied.Methods We tested the hypothesis that variants in IL-8-251 A/T was associated with susceptibility to ACS and its recurrence in a Chinese case-control study comprising 675 patients with ACS and 636 control subjects and replicated the investigation in an independent study comprising 360 cases and 360 control subjects. The plasma concentration of IL-8 was measured by enzyme-linked immunosorbent assay.Results IL-8 -251A】T poly-morphism was associated with increased susceptibility to ACS (P=0.004;OR=1.30 CI:1.12-1.53).Replication in the second study yielded similar results.IL-8 -251 A/T may affect the expression of IL-8 by the evidence that augmented IL-8 production revealed in serum of the AMI patients by ELISA. Conclusions IL-8 -251 A/T polymorphism is associated with ACS risk in Chinese Han population and An allele of IL-8- 251A/T may be an independent predictive factor.

Interleukin-17A gene variants and risk of coronary artery disease:a large angiography-based study

Background Recent studies have also revealed that interleukin(IL)-17A plays a key role in atherosclerosis and its complication,but the relationship of its common variants with coronary artery disease(CAD) has not been extensively studied.Methods We systematically screened sequence variations in the IL17A gene and designed an angiog-raphy -based case-controlled study consisting of 1031 CAD patients and 935 control subjects to investigate the association between the selected polymorphisms of IL-17A gene and CAD risk in Chinese Han population.Results Frequencies of IL17A rs8193037 GG homozygote and G allele were significantly higher in the patient group than those in the control group(P【0.001;OR=0.68;95%CI=0.54-0.85).Stratification analysis showed that the IL17A rs8193037 G allele significantly increased the risk of CAD only among male subjects (P=0.001;OR=0.63;95%CI=0.47-0.83).After adjustment for conventional risk factors,binary logistic regression analysis showed that the G allele carriers(GG +AG) had significantly increased CAD risk compared with the AA homozygotes (adjusted P【0.001;OR 0.43;95%CI,0.33- 0.58).ELISA showed augmented IL17A production in plasma of the AMI patients.Conclusions Based on our data,we speculated that the SNP rs8193037 of IL17A gene is significantly associated with CAD risk in Chinese Han population and the rs8193037 G allele which is associated with increased expression of IL17A in AMI patients may be an independent predictive factor for CAD.

MEK AND p38 MAPK-DEPENDANT PATHWAYS ARE INVOLOVED IN THE POSITIVE EFFECT OF INTERLEUKIN-6 ON HUMAN GROWTH HORMONE GENE EXPRESSION IN RAT MtT/S SOMATOTROPH CELLS

Objective To investigate the effect of interleukin-6(IL-6)on the human growth hormone(hGH)gene expression in a rat somatotropic pituitary cell line MtT/S.Methods The plasmids containing various lengths of hGH gene 5'-promoter fragments were constructed.Stably transfected MtT/S cells were created by cotransfecting the above plasmids and pcDNA3.1(+)with DMRIE-C transfection reagent.After the administration of these cells with IL-6 and/or various inhibitors of signaling transduction pathways,the luciferase activities in MtT/S cells lysis were assayed to demonstrate the effects of IL-6 on hGH gene promoter activity and possibly involved mechanism.Results The 103 U/mL IL-6 stimulated GH secretion and synthesis,and promoted the 5'-promoter activity of GH gene in stably transfected MtT/SGL cells with the action of 1.69 times above the control.Among inhibitors of signaling transduction pathways,mitogen-activated protein kinase kinase(MAPKK/MEK)inhibitor PD98059(40 μmol/L)and p38 mitogen-activated protein kinase(MAPK)inhibitor SB203580(5 μmol/L)completely blocked the stimulatory effect of IL-6.Western blot analysis further confirmed the activation of phosphorylated MEK and p38 MAPK in MtT/SGL cells.Neither over-expression of Pit-1 nor inhibition of Pit-1 expression affected IL-6 induction of hGH promoter activity.A series of deletion constructs of hGH promoter were created to identify the DNA sequence that mediated the effect of IL-6.The results showed that the stimulatory effect of IL-6 was abolished following deletion of the-196 to-132 bp fragment.Conclusions IL-6 promotes GH secretion and synthesis by rat MtT/S somatotroph cells.The stimulatory effect of IL-6 on hGH gene promoter appears to require the activation of MEK and p38 MAPK,and a fragment of promoter sequence that spans the-196 to-132 bp of the gene,but may be unlinked with Pit-1 protein.