Oral variant of acantholytic squamous cell carcinoma—Histochemical and immunohistochemical features

Acantholytic squamous cell carcinoma (ASCC) is an uncommon variant of squamous cell carcinoma (SCC). It is characterized by a combination of typical SCC and pseudoglandular structures, dyskeratotic cells and prominent acantholysis.The purpose of this study was to analyze the histochemical and immunohistochemical characteristics of the intraoral variant of ASCC. Cases of intraoral ASCC were retrieved from the English language literature. Four new cases from our files were added.In total, 35 cases were included and analyzed in this study. The mean age of the patients was 61.5 + 13 years (age range 38–92 years), with a male-to-female ratio of 1.7:1. According to the available data, histochemical and immunohistochemical stains for mucins were found to be consistently negative. E- cadherin, a marker of adherens junctions, was usually reported to be expressed in areas of “typical” (non acantholytic) SCC, but reduced in the acantholytic areas. We examined for the first time the expression of claudin 1, a marker of tight junctions, and found it to be reduced in the acantholytic areas, similar to E-cadherin. Several cases of oral ASCC also expressed vimentin and cytokeratin (CK) 19, markers associated with epithelial-mesenchymal transition. A wide range of non-epithelial markers yielded negative immunoreactions.In conclusion, ASCC is an uncommon variant of squamous cell carcinoma. The acantholytic process appears to involve reduced expression of molecular components of both adherens junctions and tight junctions. These findings could suggest a relation to the epithelial mesenchymal transition process and therefore further studies are needed in order to establish such a link and the subsequent possible impact on the clinical outcome of the patients.     

Highly expressed Claudin18.2 as a potential therapeutic target in advanced gastric signet-ring cell carcinoma (SRCC)

BackgroundAdvanced gastric signet-ring cell carcinoma (SRCC) is a specific type of malignant gastric cancer (GC) with distinct poorer survival. Claudin18.2 (CLDN18.2) is a promising neo-biomarker for the treatment of GC. Clinical trials of CLDN18.2-targeted antibody and T cell-based immunotherapy providing promising prospects for the treatment of GC. The effect of antibody therapy depended on the expression rate of CLDN18.2 has been found in clinical trials. This study aimed to determine the prevalence and the therapeutic value of CLDN18.2 in advanced gastric SRCC.MethodsExpression of CLDN18.2 in 105 formalin-fixed, paraffin-embedded (FFPE) tumor tissues was detected by immunohistochemistry (IHC) and evaluated according to FAST criteria. Next-generation sequencing (NGS) using 416 pan-cancer genes panel was performed to characterize the genomic landscape in 61 advanced gastric SRCC patients. Fisher’s exact test was used to determine gene differences in different CLDN18.2 expression levels.ResultsA total number of 105 advanced gastric SRCC samples were analyzed, of which 95.2% (100/105) were positive stained. Moderate-to-strong CLDN18.2 expression was observed in 64.8% (68/105) of all samples. In particularly, 21.0% (22/105) samples had positive staining in more than 90% tumor cells. No significance was found between CLDN18.2 expression and overall survival (OS). NGS results showed that single nucleotide variations (SNVs) could be frequently found in TP53 (26.2%), CDH1 (19.7%), MED12 (18.0%), PKHD1 (18.0%) and ARID1A (11.5%), besides, copy number variations (CNVs) were rich in NOTCH1 (18.0%) and FLT4 (9.8%) in SRCC samples. Moreover, SNVs in GRIN2A was found in 20% of the patients who had CLDN18.2 staining in <40% of tumor cells (P=0.043), indicating CLDN18.2 expression might be related to the aberration of GRIN2A in advanced gastric SRCC.ConclusionsThe highly expressed CLDN18.2 among advanced gastric SRCC patients that we found certified the value of CLDN18.2-targeted therapy in this specific type of GC. In addition, Analyses between CLDN18.2 expression and genetic abnormalities provided novel therapeutic options for advanced gastric SRCC.     

除湿胃苓汤对小鼠特异性皮炎的治疗作用及组织ERK1/2、Claudin 1表达的影响

[目的] 探究除湿胃苓汤(Chushi Weiling Decoction,CSWLD)对特异性皮炎(atopic dermatitis,AD)小鼠的皮损改善效果,以及对组织细胞外信号调节激酶1/2(extracellular signal-regulated kinase1/2,ERK1/2)、紧密连接蛋白1(Claudin 1)表达的影响。[方法]采用二硝基氟苯(dinitro fluoro benzene,DNFB)诱导建立小鼠AD模型,随机分为模型组(0.9%氯化钠溶液)和低、中、高CSWLD组(5、10、20 mL·kg-1),另设置空白组(0.9%氯化钠溶液),各组均以相应药物灌胃1周。给药结束后,评价皮损改善情况,并行炎症评分。酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法检测血清白细胞介素-33(interleukin-33,IL-33)、IL-6、IL-4、CC类趋化因子配体-5(C-C motif chemokine ligand-5,CCL-5)和CCL-2含量。取皮损部位组织,以免疫组化染色测定IL-33、肿瘤抑制素2(suppression of tumorigenicity 2,ST2)、Claudin 1含量;以定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)和免疫印迹检测ERK1/2、Claudin 1的mRNA和蛋白表达。[结果] 与空白组比较,模型组小鼠皮肤红肿、干燥、结痂明显,炎症评分增加,血清IL-33、IL-6、IL-4、CCL-5、CCL-2含量升高,组织IL-33、ST2含量增加,Claudin 1 mRNA表达降低,ERK1/2 mRNA表达升高,Claudin 1蛋白表达降低,ERK1/2磷酸化产物(phosphorylated-ERK1/2,p-ERK1/2)与ERK1/2蛋白比值增加(均P<0.01)。在给予不同剂量CSWLD后,小鼠皮损部位红肿消退,结痂脱落,症状减轻。与模型组比较,中、高CSWLD组小鼠炎症评分降低,血清IL-33、IL-6、IL-4、CCL-5、CCL-2 含量下降,组织IL-33、ST2 含量降低,Claudin 1 mRNA 表达升高,ERK1/2 mRNA 表达降低,Claudin 1蛋白表达升高,p-ERK1/2与ERK1/2蛋白比值降低(P<0.05,P<0.01);而低CSWLD组小鼠的各项指标差异无统计学意义(P>0.05)。[结论] CSWLD可下调炎性介质,并能抑制组织ERK1/2表达、促进组织Claudin 1表达,缓解AD小鼠皮损症状。     

claudin蛋白及其在急性肺损伤中的作用

claudin蛋白是肺泡上皮细胞紧密连接结构的重要组成部分,它是一个多基因家族,到目前为止已有24个家族成员,有7种claudin在肺部表达.claudin在肺泡上皮细胞中表达不同与细胞旁路通透性有关,在肺泡上皮细胞屏障中起到重要作用.本文就claudin蛋白的生理学特性以及它在急性肺损伤中的作用进行综述.     

回药扎里奴思方联合BMSCs移植对脑缺血再灌注大鼠BBB紧密连接蛋白的影响

目的观察扎里奴思方联合BMSCs移植对MCAO模型大鼠BBB上Occludin和Claudin mRNA表达的影响。方法 250只SD大鼠随机分为假手术组、模型组、扎方组、移植组和联合组,除假手术组10只外,其余各组15只;线栓法制备MCAO模型,体外全骨髓贴壁法培养及扩增BMSCs;大鼠灌胃给药[14.6 g/(kg·d)],BMSCs悬浮液经颈内动脉移植入脑(2×106/200μl);移植后1 d、3 d、7 d、14 d取材,干湿重法检测脑含水量,Real timePCR技术检测Occludin和Claudin mRNA表达。结果模型大鼠脑含水量较假手术组增加(P0.01),Occludin和Claudin mRNA表达降低(P0.01);与模型组比较,扎方、移植及联合各3 d、7 d、14 d组脑含水量降低(P0.01),各组各时间点Occludin和Claudin mRNA表达增高(P0.01);与移植组比较,扎方1 d组脑含水量降低(P0.05),3 d组Occludin mRNA表达增高(P0.01),7 d组表达降低(P0.01),扎方1 d组Claudin mRNA表达增高(P0.05),7 d、14 d组表达降低(P0.01),联合各组脑含水量均降低,以1 d、14 d明显(P0.01),各组Occludin和Claudin mRNA表达均增高(P0.01);扎方与联合组比较,联合各组脑含水量降低,以7 d、14 d组明显(P0.01,P0.05),Occludin和Claudin mRNA表达增高(P0.01);同组间比较,均以7 d组变化显著,脑含水量呈先增后减趋势,Occludin和Claudin mRNA表达呈先减后增趋势,14 d有明显改善(P0.01)。结论脑缺血再灌注损伤后BBB受损,通透性改变,脑水肿形成,以7 d最为明显;扎里奴思方和BMSCs移植均可不同程度改善脑缺血后脑水肿程度,以二者联合应用作用显著,其机制可能与干预Occludin和Claudin mRNA动态表达有关。     

度他雄胺对大鼠附睾Claudin1和β-catenin表达的影响

目的:通过度他雄胺对大鼠附睾上皮连接关键蛋白Claudin1和β-catenin表达影响的研究,探讨度他雄胺抑制生育的可能分子机制,为男性不育治疗和男性避孕提供附睾靶点。方法:16只SD雄性大鼠(3月龄,250~300 g),设度他雄胺40 mg/kg实验组和阴性对照组,每组8只,灌胃给药,1次/d,连续2周。给药结束后计算机辅助精子分析雄鼠精子活力及形态,计算交配雌鼠受孕率,ELISA法测定大鼠血清睾酮(T)和双氢睾酮(DHT)浓度,透射电镜分析附睾细胞紧密连接变化,RT-PCR分析实验组和对照组附睾上皮连接蛋白Claudin1和β-catenin基因表达变化;免疫组化和免疫印迹分析其蛋白表达变化。结果:度他雄胺可以显著抑制大鼠血清DHT浓度、精子活力和生育力,超微结构显示实验组附睾上皮细胞紧密连接出现空隙,附睾腔液体积增多。RT-PCR、免疫组化和免疫印迹显示度他雄胺可以显著下调Claudin1和β-catenin的表达。结论:度他雄胺可以显著抑制雄性大鼠生育,这可能与其降低大鼠血清DHT水平,进而抑制附睾上皮连接关键基因Claudin1和β-catenin表达和破坏附睾上皮紧密连接有关。     

Progesterone exerts neuroprotective effects by inhibiting inflammatory response after stroke

Objective and design  We evaluated the inhibitory effects of progesterone (PROG) on inflammatory response and its influence on the structure of blood–brain barrier in a permanent model of stroke. Material  One hundred and twenty adult male Sprague-Dawley rats were used in this study. Treatments  PROG was dissolved in 22.5% 2-hydroxypropyl-bcyclodextrin and given in a dose of 15 mg/kg by intraperitoneal injection 1 h after permanent occlusion of middle cerebral artery (pMCAO). Additional injections of 15 mg/kg were administered subcutaneously 6, 24, and 48 h after pMCAO. Methods  The expression of tumor necrosis factor-alpha (TNF-α) and claudin5 was measured by immunohistochemistry and western blot technique. Brain water content was determined by the dry–wet weight method. Results  TNF-α were increased, but claudin5 were reduced in vehicle-treated rats after pMCAO. PROG-treated rats showed a substantial reduction in the expression of TNF-α compared to vehicle controls. In addition, there was significant increase in the expression of claudin5 in the pMCAO rats treated with PROG compared to vehicle. Examination of the water content of the brain also revealed that administration of PROG significantly attenuated the amount of water compared to vehicle in the ipsilateral hemispheres. Conclusions  These data indicate that PROG is beneficial in this animal model, and may warrant further test in future clinical trials for human stroke.