Lactobacilli inhibit interleukin-8 production induced by Helicobacter pylori lipopolysaccharide-activated Toll-like receptor 4

AIM： To investigate the effect of Lactobacillus bulgaricus （LBG） on the Toll-like receptor 4 （TLR4） pathway and interleukin-8 （IL-8） production in SGC-7901 cells treated with Helicobacter pyloriSydney strain 1 lipopolysaccharide （HpyloriSS1-LPS）. METHODS： SGC-7901 cells were treated with HpyloriSS1-LPS in the presence or absence of pretreatment for 1 h with viable LBG or supernatant recovered from LBG culture MRS broth （LBG-s）. Cellular lysates were prepared for Western blot with anti-TLR4, anti-transforming growth factor β-activated kinase 1 （TAK1）, anti-phospho-TAK1, anti-nuclear factor κB （NF-κB）, anti-p38 mitogen-activated protein kinase （p38MAPK）, and anti-phospho-p38MAPK antibodies. The amount of IL-8 in cell culture medium was measured by ELISA. RESULTS： H pyloriSS1-LPS up-regulated the expression of TLR4, stimulated the phosphorylation of TAKI, subsequently enhanced the activation of NF- κB and the phosphorylation of p38MAPK in a time- dependent manner, leading to augmentation of IL-8 production in SGC-7901 cells. Viable LBG or LBG-s pretreatment attenuated the expression of TLR4, inhibited the phosphorylation of TAK1 and p38MAPK, prevented the activation of NF-κB, and consequently blocked IL-8 production.CONCLUSION： H py/oriSS1-LPS induces IL-8 production through activating TLR4 signaling in SGC-7901 cells and viable LBG or LBG-s prevents H pyloriSS1-LPS-mediated IL-8 production via inhibition of the TLR4 pathway.

Amino acid deletions at positions 893 and 894 of cytotoxinassociated gene A protein affect Helicobacter pylori gastric epithelial cell interactions

BACKGROUND Helicobacter pylori(H.pylori)persistently colonizes the human gastric mucosa in more than 50%of the global population,leading to various gastroduodenal diseases ranging from chronic gastritis to gastric carcinoma.Cytotoxin-associated gene A(CagA)protein,an important oncoprotein,has highly polymorphic Glu-Pro-Ile-Tyr-Ala segments at the carboxyl terminus,which play crucial roles in pathogenesis.Our previous study revealed a significant association between amino acid deletions at positions 893 and 894 and gastric cancer.AIM To investigate the impact of amino acid deletions at positions 893 and 894 on CagA function.METHODS We selected a representative HZT strain from a gastric cancer patient with amino acid deletions at positions 893 and 894.The cagA gene was amplified and mutated into cagA-NT and cagA-NE(sequence characteristics of strains from nongastric cancer patients),cloned and inserted into pAdtrack-CMV,and then transfected into AGS cells.The expression of cagA and its mutants was examined using realtime polymerase chain reaction and Western blotting,cell elongation via cell counting,F-actin cytoskeleton visualization using fluorescence staining,and interleukin-8(IL-8)secretion via enzyme-linked immunosorbent assay.RESULTS The results revealed that pAdtrack/cagA induced a more pronounced hummingbird phenotype than pAdtrack/cagA-NT and pAdtrack/cagA-NE(40.88±3.10 vs 32.50±3.17,P<0.001 and 40.88±3.10 vs 32.17±3.00,P<0.001)at 12 hours after transfection.At 24 hours,pAdtrack/cagA-NE induced significantly fewer hummingbird phenotypes than pAdtrack/cagA and pAdtrack/cagA-NT(46.02±2.12 vs 53.90±2.10,P<0.001 and 46.02±2.12 vs 51.15±3.74,P<0.001).The total amount of F-actin caused by pAdtrack/cagA was significantly lower than that caused by pAdtrack/cagA-NT and pAdtrack/cagA-NE(27.54±17.37 vs 41.51±11.90,P<0.001 and 27.54±17.37 vs 41.39±14.22,P<0.001)at 12 hours after transfection.Additionally,pAdtrack/cagA induced higher IL-8 secretion than pAdtrack/cagA-NT and pAdtrack/cagA-NE at different times after transfection.CONCLUSION Amino acid deletions at positions 893 and 894 enhance CagA pathogenicity,which is crucial for revealing the pathogenic mechanism of CagA and identifying biomarkers of highly pathogenic H.pylori.

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.

Chemokine receptor 8 expression may be linked to disease severity and elevated interleukin 6 secretion in acute pancreatitis

BACKGROUND Acute pancreatitis(AP)is an inflammatory disease,which presents with epigastric pain and is clinically diagnosed by amylase and lipase three times the upper limit of normal.The 2012 Atlanta classification stratifies the severity of AP as one of three risk categories namely,mild AP(MAP),moderately severe AP(MSAP),and severe AP(SAP).Challenges in stratifying AP upon diagnosis suggest that a better understanding of the underlying complex pathophysiology may be beneficial.AIM To identify the role of the chemokine receptor 8(CCR8),expressed by T-helper type-2 Lymphocytes and peritoneal macrophages,and its possible association to Interleukin(IL)-6 and AP stratification.METHODS This study was a prospective case-control study.A total of 40 patients were recruited from the Chris Hani Baragwanath Academic Hospital and the Charlotte Maxeke Johannesburg Academic Hospital.Bioassays were performed on 29 patients(14 MAP,11 MSAP,and 4 SAP)and 6 healthy controls as part of a preliminary study.A total of 12 mL of blood samples were collected at Day(D)1,3,5,and 7 post epigastric pain.Using multiplex immunoassay panels,real-time polymerase chain reaction(qRT-PCR)arrays,and multicolour flow cytometry analysis,immune response-related proteins,genes,and cells were profiled respectively.GraphPad Prism^(TM) software and fold change(FC)analysis was used to determine differences between the groups.P<0.05 was considered significant.RESULTS The concentration of IL-6 was significantly different at D3 post epigastric pain in both the MAP group and MSAP group with P=0.001 and P=0.013 respectively,in a multiplex assay.When a FC of 2 was applied to identify differentially expressed genes using RT2 Profiler,CCR8 was shown to increase steadily with disease severity from MAP(1.33),MSAP(38.28)to SAP(1172.45)median FC.Further verification studies using RT-PCR showed fold change increases of CCR8 in MSAP and SAP ranging from 1000 to 1000000 times when represented as Log10,compared to healthy control respectively at D3.The findings also showed differing lymphocyte and monocyte cell frequency between the groups.With monocyte population frequency as high as 70%in MSAP at D3.CONCLUSION The higher levels of CCR8 and IL-6 in the severe patients and immune cell differences compared to MAP and controls provide an avenue for exploring AP stratification to improve management.

Inhibition of Osteoarthritis in Rats by Electroporation with Interleukin-1 Receptor Antagonist

Gene therapy constitutes a promising strategy for the treatment of osteoarthritis (OA). We assessed the use of electroporation (EP) of non-viral gene vectors, and compared its efficacy with that of adeno-associated virus (AAV) vectors. EP- and AAV-mediated delivery of human interleukin-1 receptor antagonist (hIL-1Ra) was localized performed in the joints of rats following induction of OA. mRNA levels for hIL-1Ra, IL-1β, TNF-α, MMP-13 and ADAMTS-4 in the cartilage and synovial tissues were analyzed. Structural analyses of the subchondral bone at the medial femoral condyle were performed by Micro-CT after treatment. Knee joint specimens were staining with hematoxylin and eosin and Saffron O. Induction of hIL-1Ra by both EP and AAV inhibited inflammatory-induced sub-chondral bone reconstruction, and effectively suppressed IL-1β activity, as evidenced by decreased expression of MMP-13 and ADAMTS-4. Histological analyses revealed significant protection of cartilage, proteoglycan by EP and AAV. hIL-1Ra expression was similar in both the EP and AAV groups. Notably, this gene is not easier degraded transduced by EP compared with AAV. Taken together, these results show that EP offers transfection efficiency comparable to that of AAV, with the potential for longer gene expression, making EP a promising candidate for efficient non-viral delivery of OA gene therapy.

Expression of toll-like receptor 4 gene polymorphism and inflammatory factors in peripheral blood on patients with sepsis

Objective:To observe the presence of toll-like receptor 4(TLR4)gene polymorphism in the venous blood in patients with sepsis,meanwhile,to observe the expression and the diagnostic value of TLR4 mRNA,interferon-(IFN-),interleukin-23(IL-23),procalcitonin(PCT)and C-reactive protein(CRP)in patients with different degrees of sepsis.Methods:The peripheral blood samples from the subjects were collected to extract genomic DNA,gene sequencing and enzyme digestion method were applied to the detection of TLR4 Asp299Gly loci polymorphism.The expression of TLR4 mRNA in the peripheral blood was detected by reverse transcriptase polymerase chain reaction(RT-PCR).The expression levels of IFN-,IL-23,PCT and CRP were measured by enzyme linked immunosorbent assay(ELISA).Results:(1)The gene polymorphism was not present in TLR4 Asp299Gly loci;(2)The expression of TLR4 mRNA in the peripheral blood in patients with sepsis:on d1,there were statistically significant differences between the normal group and the group(APACHE II≤20),between the normal group and the group(APACHE II>20),between the group(APACHE II≤20)and the group(AAPACHE II>20)(t=5.741,14.780 and 10.500,all p<.01).APACHE II stands for Acute Physiology and Chronic Health Evaluation II.On d7,there were statistically significant differences between the normal group and the group(APACHE II≤20),between the normal group and the group(APACHE II>20),between the group(APACHE II≤20)and the group(APACHE II>20)(t=4.186,13.830 and 9.645,all p<.01);(3)The expression levels of IFN-,IL-23,PCT and CRP were obviously upregulated(all p<.01),and TLR4 was positively correlated with IFN-and IL-23(all p<.01);(4)The best cutoff value of TLR4 mRNA at baseline was 891.6μg/L,with a sensitivity of 100%and a diagnostic specificity of 57%.The best cutoff value of IFN-at baseline was 84.5μg/L,with a sensitivity of 100%and a diagnostic specificity of 57%.The best cutoff value of IL-23 at baseline was 861μg/L,with a sensitivity of 100%and a diagnostic specificity of 97%.Conclusions:(1)The Asp299Gly polymorphism is not present in TLR4 gene;(2)The expression levels of TLR4 mRNA,IFN-and IL-23 are relatively high in patients with sepsis,and will be higher with the increased severity of sepsis;(3)TLR4 mRNA,IFN-and IL-23 can be used as molecular biomarkers for the early diagnosis of sepsis.

BRAF、TP53、Pax8-PPARγ在甲状腺癌中的表达及疗效预测价值

目的探讨肿瘤蛋白P53(TP53)、丝氨酸/苏氨酸蛋白激酶(BRAF)、配对盒基因8-过氧化物酶体增殖物激活受体γ(Pax8-PPARγ)在甲状腺癌中的表达及疗效预测价值。方法将2020年4月至2022年4月该院收治的150例甲状腺癌患者纳入研究。检测患者甲状腺癌组织及癌旁组织中TP53、BRAF、Pax8-PPARγmRNA的表达水平,分析其与临床病理因素的关系,基于受试者工作特征(ROC)曲线及决策曲线分析TP53、BRAF、Pax8-PPARγmRNA表达水平预测^(131)I治疗效果的价值。结果甲状腺癌组织中的TP53、BRAF、Pax8-PPARγmRNA表达水平高于癌旁组织(P<0.05)。甲状腺癌患者淋巴结转移、肿瘤最大径、包膜浸润与TP53、BRAF、Pax8-PPARγmRNA表达水平有关(P<0.05)。采用放射性^(131)I清除术后残留的甲状腺组织(简称清甲)失败患者组织TP53、BRAF、Pax8-PPARγmRNA表达水平均高于清甲成功患者(P<0.05)。ROC曲线分析显示,TP53、BRAF、Pax8-PPARγmRNA联合预测甲状腺癌患者清甲失败的曲线下面积优于三者单独预测(P<0.05)。决策曲线显示,三者联合预测甲状腺癌清甲失败发生的净收益率优于单一预测(P<0.05)。结论TP53、BRAF、Pax8-PPARγ在甲状腺癌组织中呈高表达,联合检测有助于预测^(131)I治疗效果,为临床确定合理治疗方案及时机提供参考。

Autocrine IL-8 Contributes to Propionibacterium Acnes-induced Proliferation and Differentiation of HaCaT Cells via AKT/FOXO1/Autophagy

Objective Proprionibacterium acnes(P.acnes)-induced inflammatory responses,proliferation and differentiation of keratinocytes contribute to the progression of acne vulgaris(AV).P.acnes was found to enhance the production of interleukin-8(IL-8)by keratinocytes.This study aimed to investigate the role of IL-8 in P.acnes-induced proliferation and differentiation of keratinocytes and the underlying mechanism.Methods The P.acnes-stimulated HaCaT cell(a human keratinocyte cell line)model was established.Western blotting and immunofluorescence were performed to detect the expression of the IL-8 receptors C-X-C motif chemokine receptor 1(CXCR1)and C-X-C motif chemokine receptor 2(CXCR2)on HaCaT cells.Cell counting kit-8(CCK-8)assay,5-ethynyl-20-deoxyuridine(EdU)assay and Western blotting were performed to examine the effects of IL-8/CXCR2 axis on the proliferation and differentiation of HaCaT cells treated with P.acnes,the IL-8 neutralizing antibody,the CXCR2 antagonist(SB225002),or the CXCR1/CXCR2 antagonist(G31P).Western blotting,nuclear and cytoplasmic separation,CCK-8 assay,and EdU assay were employed to determine the downstream pathway of CXCR2 after P.acnes-stimulated HaCaT cells were treated with the CXCR2 antagonist,the protein kinase B(AKT)antagonist(AZD5363),or the constitutively active forkhead box O1(FOXO1)mutant.Finally,autophagy markers were measured in HaCaT cells following the transfection of the FOXO1 mutant or treatment with the autophagy inhibitor 3-methyladenine(3-MA).Results The expression levels of CXCR1 and CXCR2 were significantly increased on the membrane of HaCaT cells following P.acnes stimulation.The IL-8/CXCR2 axis predominantly promoted the proliferation and differentiation of P.acnes-induced HaCaT cells by activating AKT/FOXO1/autophagy signaling.In brief,IL-8 bound to its receptor CXCR2 on the membrane of keratinocytes to activate the AKT/FOXO1 axis.Subsequently,phosphorylated FOXO1 facilitated autophagy to promote the proliferation and differentiation of P.acnes-induced keratinocytes.Conclusion This study demonstrated the novel autocrine effect of IL-8 on the proliferation and differentiation of P.acnes-induced keratinocytes,suggesting a potential therapeutic target for AV.

肺癌组织PTPN6、PAX8表达水平及其与患者预后的关系

目的:探讨蛋白质酪氨酸磷酸酶非受体型6(PTPN6)、配对盒基因8抗体(PAX8)在肺癌组织中表达水平及其与患者预后的关系。方法:选取本院2019年1月至2021年1月收治的86例肺癌患者,在术中收集癌组织和癌旁组织。采用实时荧光定量PCR(qRT-PCR)检测组织中PTPN6、PAX8的mRNA表达,采用免疫组化法检测组织中PTPN6、PAX8的蛋白表达。结果:PTPN6 mRNA在肺癌组织中的表达低于癌旁组织(P<0.05);PAX8 mRNA在肺癌组织中的表达高于癌旁组织(P<0.05);肺癌组织中PTPN6蛋白阳性表达的患者3年生存率高于阴性表达者(P<0.05);PAX8蛋白阳性表达的患者3年生存率低于阴性表达者(P<0.05);淋巴结转移、肿瘤直径>3.0 cm、低分化、TNM分期为Ⅲ~Ⅳ期、PTPN6阴性表达、PAX8阳性表达为影响肺癌患者预后的危险因素(P<0.05)。结论:肺癌组织PTPN6表达水平下降,PAX8表达水平上升,可作为评估患者预后的指标。

中国荷斯坦牛白介素8受体基因编码区多态性与乳腺炎的关联分析

【目的】通过研究中国荷斯坦牛白介素8受体的亚基A和B(IL8RA和IL8RB)基因编码区多态性与乳腺炎的关联性,找到与乳腺炎相关SNPs,为乳腺炎抗性分子育种提供可用的分子标记。【方法】采用巢氏PCR、DNA测序和CRS-PCR-RFLP方法,对中国荷斯坦牛的IL8RA和IL8RB编码区的遗传多态性进行了研究,分别利用SHEsis软件和PHASE软件进行配对连锁不平衡分析和单倍型分析。【结果】找到了8个SNPs,其中5个为新SNPs,分别为IL8RA的980(G/A)、995(G/A)和1008(C/T)和IL8RB的938(C/T)、959(C/T),这8个位点均不连锁。对IL8RA和IL8RB的单倍型分析发现,单倍型频率大于0.01的有15种。各SNPs与中国荷斯坦牛体细胞评分(SCS)和产奶量的关联分析表明,IL8RA735(C/G)、816(C/A)、819(A/G)位点与SCS、产奶量相关,IL8RA的735(C/G)、816(C/A)、819(A/G)位点的C、C、G等位基因和IL8RB的938(C/T)、959(C/T)位点的T等位基因均为低SCS、高产奶量的优秀等位基因。另外,IL8RA的CCGAAC单倍型纯合个体的SCS极显著低于CAGGGT和GCAGGC单倍型纯合个体(P<0.01),而其305d产奶量极显著高于另外两种单倍型纯合子;IL8RBCT单倍型纯合个体的SCS显著低于CC、TC和TT单倍型个体,其305d产奶量显著或极显著高于其余3种单倍型。【结论】IL8RA和IL8RB的单倍型CCGAAC和CT在SCS和产奶量方面是优良的单倍型,可作为选择抗乳腺炎的分子标记。

白细胞介素-8研究进展

白细胞介素 - 8( IL- 8)在趋化性细胞因子家族中属于 C- X- C亚家族。 IL- 8是一个多种细胞来源的趋化性细胞因子 ,而且不同类型的细胞对不同的诱导剂的反应也不相同。因为由不同的实验室从不同的来源得到该因子 ,所以对于 IL- 8有许多不同的描述术语。对于 IL- 8的生理生化特性 ,目前已基本清楚。对于 IL- 8基因表达的调节 ,现认为主要是在转录水平的调控 ,并且发现其基因 5′-侧翼区的 AP- 1、NF- κB及 NF- IL- 6结合位点是调节 IL- 8启动子功能的主要顺式作用元件。 IL - 8基因的表达需要这些元件之间高度的协同以达到有效的转录。 IL - 8受体则是一个由 5 9KDa和 67KDa亚单位组成的二聚体糖蛋白 ,存在于多种类型的细胞上 ,甚至包括一些不表达 IL- 8的细胞。 IL - 8的生物学活性并不表现出种族的特异性。 IL - 8可激活中性粒细胞 ,对 T-淋巴细胞有趋化性并能刺激嗜碱性粒细胞。内皮细胞 IL- 8基因的表达除了受化学因子的调节外还受力学因素的影响。流体切应力诱导内皮细胞表达 IL - 8,可能在急性炎症和动脉粥样硬化的发生。

促甲状腺激素受体与TTF-1、Pax-8在甲状腺乳头状癌中的表达及意义

目的探讨促甲状腺激素受体(TSHR)、甲状腺特异性转录因子-1(TTF-1)、成框基因8编码的转录因子(Pax-8)在甲状腺乳头状癌(PTC)、桥本氏病(HT)合并甲状腺乳头状癌、结节性甲状腺肿(NG)与正常甲状腺组织中的表达及意义。方法收集2018年9月至2019年6月在某院内分泌科确诊并在甲状腺外科行手术治疗的甲状腺组织,包括单纯PTC 16例、HT合并PTC 24例,NG 32例及癌旁正常甲状腺组织40例。将术中获得的甲状腺组织进行脱水、透明、包埋处理,分别行TSHR、TTF-1、Pax-8的免疫组织化学染色,显微镜下观察染色结果。采用χ^2检验进行多组间率的比较,差别有统计学意义时采用χ^2分割做两两比较。结果1)TSHR、TTF-1在甲状腺组织的强阳性表达在4组间的差别均有统计学意义(P<0.001,P=0.004),Pax-8在甲状腺组织的强阳性表达在4组间的差别无统计学意义(P=0.642)。2)调整检验水准α′=0.008,进一步两两比较发现:TSHR在PTC组的强阳性表达低于在NG组与癌旁正常组织(P<0.001,P=0.002),在HT合并PTC组的强阳性表达低于在NG组(P=0.001),TTF-1在HT合并PTC组的强阳性表达低于在NG组(P=0.002);其他组两两比较均无统计学意义(P>0.008)。3)在全部PTC组织中,TSHR、TTF-1的阳性表达与性别、年龄、肿瘤直径、淋巴结转移等均无关;Pax-8的阳性表达在有淋巴结转移者低于无淋巴结转移者(P=0.030),与性别、年龄、肿瘤直径无关。结论TSHR与TTF-1在PTC中的强阳性表达低于NG和癌旁正常甲状腺组织,Pax-8在不同类型甲状腺组织中表达无差别。Pax-8在PTC组织中的低表达易出现淋巴结转移。

TLR8基因多态性与中国东北地区汉族肺结核病的关联性分析

目的:探讨Toll样受体8(TLR8)基因的4个位点单核苷酸多态性(SNP)与我国东北地区汉族人群结核病的关联性,分析其基因型频数分布在肺结核病与其他肺病间的差异。方法:采用病例-对照的研究方法。选择中国东北地区368例汉族肺结核病患者(病例组)和355例汉族其他肺病患者(对照组)为研究对象。应用连接酶特异检测技术检测2组患者TLR8基因4个位点rs3764880、rs3761624、rs3788935和rs3764879的基因型,比较2组患者等位基因频数分布差异,计算其危险系数(OR),并比较不同性别患者组间基因型频数分布的差异。结果 :TLR8基因rs3764880、rs3761624、rs3788935和rs3764879位点等位基因型频数在2组间分布差异无统计学意义(P>0.05);不同性别的2组患者TLR8基因4个位点等位基因频数分布差异无统计学意义(P>0.05)。结论:TLR8基因位点rs3764880、rs3761624、rs3788935和rs3764879的SNP在我国东北地区汉族肺结核患者与其他肺病患者中的分布相同,提示上述TLR8基因4个位点的SNP与结核病的发生无关联性。

Carcinogenesis of nasopharyngeal carcinoma:an alternate hypothetical mechanism

Current proposed mechanisms implicate both early and latent Epstein-Barr virus(EBV)infection in the carcinogenic cascade,whereas epidemiological studies have always associated nasopharyngeal carcinoma(NPC)with early childhood EBV infection and with chronic ear,nose,and sinus conditions.Moreover,most patients with NPC present with IgA antibody titers to EBV capsid antigen(VCA-lgA),which can precede actual tumor presentation by several years.If early childhood EBV infection indeed constitutes a key event in NPC carcinogenesis,one would have to explain the inability to detect the virus in normal nasopharyngeal epithelium of patients at a high risk for EBV infection.It is perhaps possible that EBV resides within the salivary glands,instead of the epithelium,during latency.This claim is indirectly supported by observations that the East Asian phenotype shares the characteristics of an increased susceptibility to NPC and immature salivary gland morphogenesis,the latter of which is influenced by the association of salivary gland morphogenesis with an evolutionary variant of the human ectodysplasin receptor gene(EDAR),EDARV370A.Whether the immature salivary gland represents a more favorable nidus for EBV is uncertain,but in patients with infectious mononucleosis,EBV has been isolated in this anatomical organ.The presence of EBV-induced lymphoepitheliomas in the salivary glands and lungs further addresses the possibility of submucosal spread of the virus.Adding to the fact that the fossa of Rosen Muller contains a transformative zone active only in the first decade of life,one might be tempted to speculate the possibility of an alternative carcinogenic cascade for NPC that is perhaps not dissimilar to the model of human papillomavirus and cervical cancer.

Interleukin-1 and TNF-α polymorphisms and Helicobacter pylori in a Brazilian Amazon population

AIM： To study the association between Interleukin-1 （IL-1） and tumor necrosis factor （TNF）-α polymorphisms, infection by Helicobacter pylori （H pylori） and the development of gastrointestinal diseases.METHODS： Genomic DNA was extracted from the peripheral blood of 177 patients with various gastrointestinal diseases and from 100 healthy volunteers. The polymorphisms in IL-1β and TNF-α genes were analyzed using the polymerase chain reactionrestriction fragment length polymorphism method （PCRRFLP） and those from IL-1RN with PCR. The presence of infection due to H pylori and the presence of the CagA toxin were detected by serology. The histopathological parameters in the gastric biopsies of the patients were according to the Sydney classification.RESULTS： A comparison of the frequencies of the different polymorphisms studied among the patients and the control group demonstrated that the allele IL- 1RN＊2 was more frequent among patients with gastric ulcers and adenocarcinoma. Carriers of the allele IL- RN＊2 and those with reactive serology for anti-CagA IgG had a greater risk of developing peptic ulcer and gastric adenocarcinoma, as well as a higher degree of inflammation and neutrophilic activity in the gastricCONCLUSION： Our results indicate a positive association between IL-1RN gene polymorphism and infection by positive H pylori CagA strains and the development of gastric ulcers and adenocarcinoma.

Inflammatory cytokine gene polymorphisms increase the risk of atrophic gastritis and intestinal metaplasia

AIM: To investigate the effects of interleukin-8 (IL-8 ), macrophage migration inhibitory factor (MIF ) gene polymorphisms, Helicobacter pylori (H. pylori ) infection, on the risk of developing severe chronic atrophic gastritis (SCAG) and intestinal metaplasia (IM). METHODS: A total of 372 cases were selected from a cohort study in Linqu County, a high risk area for gastric cancer (GC) in northern China. To obtain a sufficient group size, patients with normal or superficial gastritis were included. Based on an average follow-up period of 56 mo, the 372 cases were divided into no progres-sion group (no histological progression from normal or superficial gastritis, n = 137), group Ⅰ (progressed from normal or superficial gastritis to SCAG, n = 134) and group Ⅱ (progressed from normal or superficial gastritis to IM, n = 101). IL-8 , MIF gene polymorphisms were detected by polymerase chain reaction-based denaturing high-performance liquid chromatography analysis and DNA sequencing. RESULTS: An increased risk of SCAG was found in subjects with IL-8-251 AA genotype [odds ratio (OR) = 2.62, 95% CI: 1.23-5.72] or IL-8-251 A allele carriers (AA + AT) (OR = 1.81, 95% CI: 1.06-3.09). An elevated risk of IM was found in subjects with IL-8-251 AT genotype (OR = 2.27, 95% CI: 1.25-4.14) or IL-8-251 A allele carriers (OR = 2.07, 95% CI: 1.16-3.69). An increased risk of SCAG was found in subjects with MIF-173 GC genotype (OR = 2.36, 95% CI: 1.38-4.02) or MIF-173 C allele carriers (GC + CC) (OR = 2.07, 95% CI: 1.21-3.55). An elevated risk of IM was found in subjects with MIF-173 CC genotype (OR = 2.27, 95% CI: 1.16-4.46) or MIF-173 C allele carriers (OR = 3.84, 95% CI: 1.58-9.34). The risk of SCAG and IM was more evident in subjects carrying IL-8-251 A allele (OR = 6.70, 95% CI: 1.29-9.78) or MIF-173 C allele (OR = 6.54, 95% CI: 2.97-14.20) and positive for H. pylori infection. CONCLUSION: IL-8-251 and MIF-173 gene polymorphisms are significantly associated with the risk of SCAG and IM in a population with a high risk of GC in Linqu County, Shandong Province, China.

Correlation of IL-8 with induction, progression and metastatic potential of colorectal cancer

AIM： To investigate the expression profile of IL-8 in inflammatory and malignant colorectal diseases to evaluate its potential role in the regulation of colorectal cancer （CRC） and the development of colorectal liver metastases （CRLM）.METHODS： IL-8 expression was assessed by quantitative real-time PCR （Q-RT-PCR） and the enzyme-linked immunosorbent assay （ELISA） in resected specimens from patients with ulcerative colitis （UC, n = 6） colorectal adenomas （CRA, n = 8）, different stages of colorectal cancer （n = 48） as well as synchronous and metachronous CRLM along with their corresponding primary colorectal tumors （n = 16）.RESULTS： IL-8 mRNA and protein expression was significantly up-regulated in all pathological colorectal entities investigated compared with the corresponding neighboring tissues. However, in the CRC specimens IL-8 revealed a significantly more pronounced overexpression in relation to the CRA and UC tissues with an average 30-fold IL-8 protein up-regulation in the CRC specimens in comparison to the CRA tissues. Moreover, [L-8 expression revealed a close correlation with tumor grading. Most interestingly, IL-8 up-regulation was most enhanced in synchronous and metachronous CRLM, if compared with the corresponding primary CRC tissues. Herein, an up to 80-fold IL-8 overexpression in individual metachronous metastases compared to normal tumor neighbor tissues was found.CONCLUSION： Our results strongly suggest an association between IL-8 expression, induction and progression of colorectal carcinoma and the development of colorectal liver metastases.

Argon preconditioning protects neuronal cells with a Toll-like receptor-mediated effect

The noble gas argon has the potential to protect neuronal cells from cell death.So far,this effect has been studied in treatment after acute damage.Preconditioning using argon has not yet been investigated.In this study,human neuroblastoma SH-SY5Y cells were treated with different concentrations of argon(25%,50%,and 74%;21%O_(2),5%CO_(2),balance nitrogen)at different time intervals before inflicting damage with rotenone(20μM,4 hours).Apoptosis was determined by flow cytometry after annexin V and propidium iodide staining.Surface expressions of Toll-like receptors 2 and 4 were also examined.Cells were also processed for analysis by western blot and qPCR to determine the expression of apoptotic and inflammatory proteins,such as extracellular-signal regulated kinase(ERK1/2),nuclear transcription factor-κB(NF-κB),protein kinase B(Akt),caspase-3,Bax,Bcl-2,interleukin-8,and heat shock proteins.Immunohistochemical staining was performed for TLR2 and 4 and interleukin-8.Cells were also pretreated with OxPAPC,an antagonist of TLR2 and 4 to elucidate the molecular mechanism.Results showed that argon preconditioning before rotenone application caused a dose-dependent but not a time-dependent reduction in the number of apoptotic cells.Preconditioning with 74%argon for 2 hours was used for further experiments showing the most promising results.Argon decreased the surface expression of TLR2 and 4,whereas OxPAPC treatment partially abolished the protective effect of argon.Argon increased phosphorylation of ERK1/2 but decreased NF-κB and Akt.Preconditioning inhibited mitochondrial apoptosis and the heat shock response.Argon also suppressed the expression of the pro-inflammatory cytokine interleukin-8.Immunohistochemistry confirmed the alteration of TLRs and interleukin-8.OxPAPC reversed the argon effect on ERK1/2,Bax,Bcl-2,caspase-3,and interleukin-8 expression,but not on NF-κB and the heat shock proteins.Taken together,argon preconditioning protects against apoptosis of neuronal cells and mediates its action via Toll-like receptors.Argon may represent a promising therapeutic alternative in various clinical settings,such as the treatment of stroke.

WGCNA Reveals Key Roles of IL8 and MMP-9 in Progression of Involvement Area in Colon of Patients with Ulcerative Colitis

Ulcerative colitis （UC） is a chronic inflammatory disease and its involvement area in colon is influenced by a complex network of gene interactions. We analyzed the weighted gene co-expression networks in mieroarray dataset from colonic mucosa of patients with UC and identified one gene co-expression module that was highly associated with the progression of involved area in UC colon （Pearson coefficient=0.81, P〈0.0001）. In total, 523 hub genes in this module were found to be involved in immune system process after enrichment analysis in Gene Ontology. By the STRING and Cytoscape analysis, we observed that interleukin-8 （IL- 8） and matrix metalloproteinase-9 （MMP-9） were centered in the network of hub genes. We then detected the expression of IL-8 and MMP-9 in mucosa from left-sided colon of patients using quantitative PCR and immunofluorescence assay respectively. Both quantitative PCR and immunofluorescence assay revealed the expression levels of IL-8 and MMP-9 were significantly different among the healthy controls, left-sided colitis group and pancolitis group （P〈0.05）. IL-8 and MMP-9 were detected with an enhanced expression in pancolitis as compared with leftsided colitis and healthy controls, respectively （P〈0.05）. This study demonstrates that immune system process is indispensable in the progression of disease in colon, and identifies that IL-8 and MMP-9 play potential critical roles for the progression.