CDX2 protein expression compared to alcian blue staining in the evaluation of esophageal intestinal metaplasia

AIM:To compare the sensitivity and specificity of CDX2 and alcian blue(AB) p H 2.5 staining in identifying esophageal intestinal metaplasia.METHODS:One hundred and ninty-nine biopsies from 186 patients were retrospectively reviewed and categorized as Barrett's esophagus(BE)(n =108); non-Barrett's esophagus(NBE)(n =48); columnar blue cells(CB) and esophageal glands(EG)(n =43).The biopsies were stained with AB and immunostained for CDX2 using a mouse monoclonal antibody from Biogenex(clone CDX2-88) and the Ventana Discovery X automated immunostainer.The positive and negative predictive value of each group was used to determine the predictive power of CDX2 and AB in diagnosing intestinal metaplasia.RESULTS:All of the 108 BE biopsies(100%) were positive for AB and 102 of them(94.4%) were positive for CDX2.The six BE patients(5.6%) who failed to stain with CDX2 were found to have lost the focus of intestinal metaplasia upon deeper sectioning for immunostaining.Both AB and CDX2 were negative in 43 out of 48(89.6%) NBE cases.Five NBE patients(10.4%) were falsely positive for AB due to the presence of EG and CB in these biopsies.These cases were all CDX2 negative.In addition,5 AB negative NBE were found to be CDX2 positive.Based on these results the CDX2 immunostain had similar sensitivity but higher specificity(100% vs about 91%) than AB in detecting intestinal type metaplasia in these samples.Our data shows that CDX2 has a better PPV in detecting intestinal metaplasia as compared to AB(95.6% vs 71.5%,respectively).CONCLUSION:CDX2 has a better positive predictive value than AB in detecting intestinal metaplasia.CDX2may be useful when challenged by gastro-esophageal biopsies containing mimikers of BE.

Tousled-like kinase 1 promotes gastric cancer progression by regulating the tumor growth factor-beta signaling pathway

BACKGROUND The role of Tousled-like kinase 1(TLK1)in in gastric cancer(GC)remains unclear.AIM To investigate the expression,biological function,and underlying mechanisms of TLK1 in GC.METHODS We measured TLK1 protein expression levels and localized TLK1 in GC cells and tissues by western blot and immunofluorescence,respectively.We transfected various GC cells with lentiviruses to create TLK1 overexpression and knockdown lines and established the functional roles of TLK1 through in vitro colony formation,5-ethynyl-2`-deoxyuridine,and Transwell assays as well as flow cytometry.We applied bioinformatics to elucidate the signaling pathways associated with TLK1.We performed in vivo validation of TLK1 functions by inducing subcutaneous xenograft tumors in nude mice.RESULTS TLK1 was significantly upregulated in GC cells and tissues compared to their normal counterparts and was localized mainly to the nucleus.TLK1 knockdown significantly decreased colony formation,proliferation,invasion,and migration but increased apoptosis in GC cells.TLK1 overexpression had the opposite effects.Bioinformatics revealed,and subsequent experiments verified,that the tumor growth factor-beta signaling pathway was implicated in TLK1-mediated GC progression.The in vivo assays confirmed that TLK1 promotes tumorigenesis in GC.CONCLUSION The findings of the present study indicated that TLK1 plays a crucial role in GC progression and is,therefore,promising as a therapeutic target against this disease.

LPA_(1) antagonist-derived LNPs deliver A20 mRNA and promote anti-fibrotic activities

Activated fibroblasts are major mediators of pulmonary fibrosis.Fibroblasts are generally found in the connective tissue but upon activation can generate excess extracellular matrix(ECM)in the lung interstitial section.Therefore,fibroblasts are one of the most targeted cells for treating idiopathic pulmonary fibrosis(IPF).Here,we develop an anti-fibrotic platform that can modulate both the lysophosphatidic acid receptor 1(LPA_(1))and the inflammatory pathway through tumor necrosis factorα-induced protein 3(TNFAIP3,also known as A20)in fibroblasts.First,we synthesized a series of LPA_(1) antagonists,AM095 and AM966,derived amino lipids(LA lipids)which were formulated into LA-lipid nanoparticles(LA-LNPs)encapsulating mRNA.Specifically,LA5-LNPs,with AM966 head group and biodegradable acetal lipid tails,showed efficient A20 mRNA delivery to lung fibroblasts in vitro(80.2%±1.5%)and ex vivo(17.2%±0.4%).When treated to primary mouse lung fibroblasts(MLF),this formulation inhibited fibroblast migration and collagen production,thereby slowing the progression of IPF.Overall,LA5-LNPs encapsulated with A20 mRNA is a novel platform offering a potential approach to regulate fibroblast activation for the treatment of IPF.

IFN-γ^(+) cytotoxic CD4^(+) T lymphocytes are involved in thepathogenesis of colitis induced by IL-23 and the foodcolorant Red 40

The food colorant Red 40 is an environmental risk factor for colitis development in mice with increased expression of interleukin(IL)-23.This immune response is mediated by CD4^(+)T cells,but mechanistic insights into how these CD4^(+)T cells trigger andperpetuate colitis have remained elusive.Here,using single-cell transcriptomic analysis,we found that several CD4^(+)T-cell subsetsare present in the intestines of colitic mice,including an interferon(IFN)-γ-producing subset.In vivo challenge of primed mice withRed 40 promoted rapid activation of CD4^(+)T cells and caused marked intestinal epithelial cell(IEC)apoptosis that was attenuated bydepletion of CD4^(+)cells and blockade of IFN-γ.Ex vivo experiments showed that intestinal CD4^(+)T cells from colitic mice directlypromoted apoptosis of IECs and intestinal enteroids.CD4^(+)T cell-mediated cytotoxicity was contact-dependent and required FasL,which promoted caspase-dependent cell death in target IECs.Genetic ablation of IFN-γconstrained IL-23-and Red 40-inducedcolitis development,and blockade of IFN-γinhibited epithelial cell death in vivo.These results advance the understanding of themechanisms regulating colitis development caused by IL-23 and food colorants and identify IFN-γ^(+)cytotoxic CD4^(+)T cells as a newpotential therapeutic target for colitis.

Gene expression profiles of the one-carbon metabolism pathway

One-carbon metabolism plays a critical role in both DNA methylation and DNA synthesis. Accumulating evidence has shown that interruptions of this pathway are associated with many disease outcomes including cardiovascular diseases and cancers. Mechanistic studies have been performed on genetic polymorphisms involved in one-carbon metabolism. However, expression profiles of these inter-related genes are not well-known. In this study, we examined the gene expression profiles of 11 one-carbon metabolizing genes by quantifying the mRNA level of the lymphocyte among 54 healthy individuals and explored the correlations of these genes. We found these genes were expressed in lymphocytes at moderate levels and showed significant inter-person variations, We also applied principle component analysis to explore potential patterns of expression. The components identified by the program agreed with existing knowledge about one-carbon metabolism. This study helps us better understand the biological functions of one-carbon metabolism.

Biomaterials for in situ cell therapy

Cell therapy has revolutionized the treatment of various diseases,such as cancers,genetic disorders,and autoimmune diseases.Currently,most cell therapy products rely on ex vivo cell engineering,which requires sophisticated manufacturing processes and poses safety concerns.The implementation of in situ cell therapy holds the potential to overcome the current limitations of cell therapy and provides a broad range of applications and clinical feasibility in the future.A variety of biomaterials have been developed to improve the function and target delivery to specific cell types due to their excellent biocompatibility,tunable properties,and other functionalities,which provide a reliable method to achieve in vivo modulation of cell reprogramming.In this article,we summarize recent advances in biomaterials for in situ cell therapy including T cells,macrophages,dendritic cells,and stem cells reprogramming leveraging lipid nanoparticles,polymers,inorganic materials,and other biomaterials.Finally,we discuss the current challenges and future perspectives of biomaterials for in situ cell therapy.

One-carbon metabolism and breast cancer: an epidemiological perspective

One-carbon metabolism is a network of biological reactions that plays critical role in DNA methylation and DNA synthesis, and in turn, facilitates the cross-talk between genetic and epigenetic processes. Genetic polymorphisms and supplies of cofactors （e.g. folate, vitamins B） involved in this pathway have been shown to influence cancer risk and even survival. In this review, we summarized the epidemiological evidence for one-carbon metabolism, from both genetics and lifestyle aspects, in relation to breast cancer risk. We also discussed this pathway in relation to breast cancer survival and the modulation of one-carbon polymorphism in chemotherapy. Emerging evidence on modulation of DNA methylation by one-carbon metabolism suggests that disruption of epigenome might have been the underlying mechanism. More results are expected and will be translated to guidance to the general population for disease prevention as well as to clinicians for treatment and management of the disease.

The innate immune response to allotransplants: mechanisms and therapeutic potentials

Surgical trauma and ischemia reperfusion injury(IRI)are unavoidable aspects of any solid organ transplant procedure.They trigger a multifactorial antigen-independent inflammatory process that profoundly affects both the early and long-term outcomes of the transplanted organ.The injury associated with donor organ procurement,storage,and engraftment triggers innate immune activation that inevitably results in cell death,which may occur in many different forms.Dying cells in donor grafts release damage-associated molecular patterns(DAMPs),which alert recipient innate cells,including macrophages and dendritic cells(DCs),through the activation of the complement cascade and toll-like receptors(TLRs).The long-term effect of inflammation on innate immune cells is associated with changes in cellular metabolism that skew the cells towards aerobic glycolysis,resulting in innate immune cell activation and inflammatory cytokine production.The different roles of proinflammatory cytokines in innate immune activation have been described,and these cytokines also stimulate optimal T-cell expansion during allograft rejection.Therefore,early innate immune events after organ transplantation determine the fate of the adaptive immune response.In this review,we summarize the contributions of innate immunity to allograft rejection and discuss recent studies and emerging concepts in the targeted delivery of therapeutics to modulate the innate immune system to enhance allograft survival.

Cell Rep：研究发现衰老和癌症相关的关键蛋白

伦敦大学玛丽皇后学院（UQML）的一项早期研究发现了衰老细胞中一个蛋白此前未知的一种功能。研究人员希望这项发现在未来可以帮助开发出治疗衰老和早期癌症的新疗法。我们身体的组织和器官由大量细胞组成，这些细胞一起协作帮助机体行使正常的功能。