Background:Th9 cells are a newly identified subpopulation of CD4+helper T cells.Immune checkpoint inhibitors have emerged as a central focus in the realm of gastric cancer immunotherapy.However,the comprehension of the alterations within Th9 cells and the expression patterns of immune checkpoint molecules on their surface among gastric cancer patients remain limited.Aims:To investigate the expression and clinical significance of immune checkpoint molecules on Th9 cells in the peripheral blood of gastric cancer patients.Methods:A total of 74 patients diagnosed with gastric cancer and initially treated at the First Affiliated Hospital of Soochow University between June 2021 and September 2022 constituted the case group,while 34 healthy individuals who underwent physical examination during the same period served as the control group.The proportion of Th9 cells in the peripheral blood mononuclear cells(PBMCs)and the surface expression of PD-1,TIM-3,B7-H3 and B7-H4 were assessed using flow cytometry.Subsequently,correlations of their expression levels and the clinicopathological characteristics and prognosis of gastric cancer patients were preliminarily analyzed.Results:When compared to the control group,the proportion of Th9 cells in PBMCs of gastric cancer patients was significantly decreased,while the surface expression levels of PD-1,TIM-3,B7-H3,and B7-H4 were notably elevated(all P<0.05).Positive correlations were observed between TIM-3 and B7-H3,TIM-3 and B7-H4,as well as B7-H3 and B7-H4 on Th9 cells(all P<0.05).Gastric cancer patients aged 70 years or older demonstrated significantly higher levels of PD-1 expression on Th9 cells(P<0.05).Furthermore,low PD-1 expression was associated with a significantly prolonged progression-free survival compared to high expression(27.55 months vs.19.17 months,P<0.05).Univariate and multivariate Cox regression analysis confirmed that high PD-1 expression was an independent risk factor for postoperative disease progression in gastric cancer patients(HR=3.398,95%CI:1.278-9.033,P=0.014).Conclusions:Peripheral blood Th9 cells,along with their surface immune checkpoint molecules,are hypothesized to exert a significant influence on the initiation and progression of gastric cancer.Alterations in PD-1 expression on Th9 cells might be intricately linked with the prognosis of gastric cancer patients.

Objective To examine the expression pattern of apoptosis signal-regulating kinase 1(ASK1)in the intestinal tissues of patients with Crohn's disease(CD),and analyze its mechanistic impact on intestinal epithelial barrier function and inflammatory responses.Methods Ileal tissue samples from Crohn's disease patients and healthy controls were collected.ASK1 protein level was assessed by immunohistochemistry,and its relationship with the Crohn's disease activity index(CDAI)was analyzed.A mouse model of acute colitis was constructed using TNBS,and subjected to qRT-PCR,Western blotting and immunohistochemistry for ASK1 expression,and the association between ASK1 expression and the disease activity index was examined.Lentivirus transfection was employed to create stable Caco-2 cell lines with altered ASK1 expression,and the intestinal barrier integrity and inflammation were assessed by measuring transepithelial electrical resistance(TEER),FITC-dextran leakage,and IL-6,IL-1β levels.Furthermore,the effects of ASK1 expression on Krüppel-like factor 4(KLF4)levels was examined using qRT-PCR and Western blotting.Results ASK1 was highly expressed in the ileum of CD patients and positively correlated with CDAI.In a TNBS-induced mouse model of acute colitis,ASK1 expression was up-regulated and positively correlated with DAI.Inflammation-induced ASK1 overexpression weakened the Caco-2 cell intestinal barrier,whereas ASK1 knockdown strengthened it.Moreover,ASK1 had the capability to enhance the expression of inflammatory factors.Additionally,ASK1 knockdown increased KLF4 expression,while overexpression decreased it,indicating a negative correlation between ASK1 and KLF4.Conclusion ASK1 expression is notably higher in CD and positively correlates with disease activity.ASK1 can influence intestinal barrier integrity and inflammatory factor expression,possibly through its impact on KLF4 expression.

Objective To examine the expression pattern of apoptosis signal-regulating kinase 1(ASK1)in the intestinal tissues of patients with Crohn's disease(CD),and analyze its mechanistic impact on intestinal epithelial barrier function and inflammatory responses.Methods Ileal tissue samples from Crohn's disease patients and healthy controls were collected.ASK1 protein level was assessed by immunohistochemistry,and its relationship with the Crohn's disease activity index(CDAI)was analyzed.A mouse model of acute colitis was constructed using TNBS,and subjected to qRT-PCR,Western blotting and immunohistochemistry for ASK1 expression,and the association between ASK1 expression and the disease activity index was examined.Lentivirus transfection was employed to create stable Caco-2 cell lines with altered ASK1 expression,and the intestinal barrier integrity and inflammation were assessed by measuring transepithelial electrical resistance(TEER),FITC-dextran leakage,and IL-6,IL-1β levels.Furthermore,the effects of ASK1 expression on Krüppel-like factor 4(KLF4)levels was examined using qRT-PCR and Western blotting.Results ASK1 was highly expressed in the ileum of CD patients and positively correlated with CDAI.In a TNBS-induced mouse model of acute colitis,ASK1 expression was up-regulated and positively correlated with DAI.Inflammation-induced ASK1 overexpression weakened the Caco-2 cell intestinal barrier,whereas ASK1 knockdown strengthened it.Moreover,ASK1 had the capability to enhance the expression of inflammatory factors.Additionally,ASK1 knockdown increased KLF4 expression,while overexpression decreased it,indicating a negative correlation between ASK1 and KLF4.Conclusion ASK1 expression is notably higher in CD and positively correlates with disease activity.ASK1 can influence intestinal barrier integrity and inflammatory factor expression,possibly through its impact on KLF4 expression.

Background:Th9 cells are a newly identified subpopulation of CD4+helper T cells.Immune checkpoint inhibitors have emerged as a central focus in the realm of gastric cancer immunotherapy.However,the comprehension of the alterations within Th9 cells and the expression patterns of immune checkpoint molecules on their surface among gastric cancer patients remain limited.Aims:To investigate the expression and clinical significance of immune checkpoint molecules on Th9 cells in the peripheral blood of gastric cancer patients.Methods:A total of 74 patients diagnosed with gastric cancer and initially treated at the First Affiliated Hospital of Soochow University between June 2021 and September 2022 constituted the case group,while 34 healthy individuals who underwent physical examination during the same period served as the control group.The proportion of Th9 cells in the peripheral blood mononuclear cells(PBMCs)and the surface expression of PD-1,TIM-3,B7-H3 and B7-H4 were assessed using flow cytometry.Subsequently,correlations of their expression levels and the clinicopathological characteristics and prognosis of gastric cancer patients were preliminarily analyzed.Results:When compared to the control group,the proportion of Th9 cells in PBMCs of gastric cancer patients was significantly decreased,while the surface expression levels of PD-1,TIM-3,B7-H3,and B7-H4 were notably elevated(all P<0.05).Positive correlations were observed between TIM-3 and B7-H3,TIM-3 and B7-H4,as well as B7-H3 and B7-H4 on Th9 cells(all P<0.05).Gastric cancer patients aged 70 years or older demonstrated significantly higher levels of PD-1 expression on Th9 cells(P<0.05).Furthermore,low PD-1 expression was associated with a significantly prolonged progression-free survival compared to high expression(27.55 months vs.19.17 months,P<0.05).Univariate and multivariate Cox regression analysis confirmed that high PD-1 expression was an independent risk factor for postoperative disease progression in gastric cancer patients(HR=3.398,95%CI:1.278-9.033,P=0.014).Conclusions:Peripheral blood Th9 cells,along with their surface immune checkpoint molecules,are hypothesized to exert a significant influence on the initiation and progression of gastric cancer.Alterations in PD-1 expression on Th9 cells might be intricately linked with the prognosis of gastric cancer patients.

Inflammatory bowel disease(IBD)is a chronic inflammatory disease of the gastrointestinal tract that includes Crohn's disease and ulcerative colitis.IBD may be caused by complex interactions between genetic susceptibility,environmental factors,and alterations in the gut microbiota,resulting in dysregulated innate and adaptive immune responses.Recent studies have identified macrophages in the intestinal inflammatory response as having the plasticity to not only regulate inflammation,but also to promote tissue repair and healing.As aberrant macrophage polarization occurs during the development of IBD,the balance between the phenotype and function of pro-inflammatory M1 and anti-inflammatory M2 macrophages is regulated by extracellular and intracellular stimuli,and this process is therefore expected to be a potential target for new therapeutic approaches.This article reviewed the progress of research on macrophage polarization in IBD.

Background:The incidence of colorectal cancer(CRC)remains high in China.ITGBL1,an intergrin-like molecule,may play a significant role in the tumorigenesis and progression of CRC.Aims:To explore the expression of ITGBL1 in CRC tissues,and its clinical significance and effect on the biological function of CRC cells.Methods:Fifty-five paraffin embedding cancerous and paracancerous tissues from CRC patients undergoing surgical treatment were collected.Protein expression of ITGBL1 was detected by immunohistochemistry.The relationship between ITGBL1 expression and the clinicopathological characteristics of the CRC patients was analyzed.Furthermore,the sh-ITGBL1 lentiviral system was used to construct ITGBL1 knockdown stably-transformed HCT116 and RKO cells,then the ITGBL1 protein expression was detected by Western blotting.Effects of ITGBL1 knockdown on the proliferation of CRC cells were analyzed by CCK-8 assay and colony formation assay.Results:ITGBL1 was highly expressed in CRC tissues and cells.Expression of ITGBL1 was positively correlated with the infiltration depth,lymph node metastasis and the clinical stage of CRC(all P＜0.05),whereas no correlations were found between ITGBL expression and the gender,age,tumor size,tumor differentiation,and distant metastasis of CRC patients.Compared with the control group(sh-NC),ITGBL1 expression was significantly decreased in ITGBL1 knockdown(sh-ITGBL1)HCT116 and RKO cells,and the cell proliferation was significantly inhibited(all P＜0.05).Conclusions:Highly expressed ITGBL1 may be associated with the occurrence and development of CRC.ITGBL1 knockdown exerts a significant inhibitory effect on the proliferation of CRC cells.Therefore,ITGBL1 might be a potential biomarker,which is expected to be a new target for diagnosis and treatment of CRC.

Background:The incidence of colorectal cancer(CRC)remains high in China.ITGBL1,an intergrin-like molecule,may play a significant role in the tumorigenesis and progression of CRC.Aims:To explore the expression of ITGBL1 in CRC tissues,and its clinical significance and effect on the biological function of CRC cells.Methods:Fifty-five paraffin embedding cancerous and paracancerous tissues from CRC patients undergoing surgical treatment were collected.Protein expression of ITGBL1 was detected by immunohistochemistry.The relationship between ITGBL1 expression and the clinicopathological characteristics of the CRC patients was analyzed.Furthermore,the sh-ITGBL1 lentiviral system was used to construct ITGBL1 knockdown stably-transformed HCT116 and RKO cells,then the ITGBL1 protein expression was detected by Western blotting.Effects of ITGBL1 knockdown on the proliferation of CRC cells were analyzed by CCK-8 assay and colony formation assay.Results:ITGBL1 was highly expressed in CRC tissues and cells.Expression of ITGBL1 was positively correlated with the infiltration depth,lymph node metastasis and the clinical stage of CRC(all P＜0.05),whereas no correlations were found between ITGBL expression and the gender,age,tumor size,tumor differentiation,and distant metastasis of CRC patients.Compared with the control group(sh-NC),ITGBL1 expression was significantly decreased in ITGBL1 knockdown(sh-ITGBL1)HCT116 and RKO cells,and the cell proliferation was significantly inhibited(all P＜0.05).Conclusions:Highly expressed ITGBL1 may be associated with the occurrence and development of CRC.ITGBL1 knockdown exerts a significant inhibitory effect on the proliferation of CRC cells.Therefore,ITGBL1 might be a potential biomarker,which is expected to be a new target for diagnosis and treatment of CRC.

BACKGROUND: Cancer stem-like cells (CSCs) are a small subset of cells in tumors that exhibit self-renewal and differentiation properties. CSCs play a vital role in tumor formation, progression, relapse, and therapeutic resistance. B7-H3, an immunoregulatory protein, has many protumor functions. However, little is known about the mechanism underlying the role of B7-H3 in regulating gastric cancer (GC) stemness. Our study aimed to explore the impacts of B7-H3 on GC stemness and its underlying mechanism. METHODS: GC stemness influenced by B7-H3 was detected both in vitro and in vivo . The expression of stemness-related markers was examined by reverse transcription quantitative polymerase chain reaction, Western blotting, and flow cytometry. Sphere formation assay was used to detect the sphere-forming ability. The underlying regulatory mechanism of B7-H3 on the stemness of GC was investigated by mass spectrometry and subsequent validation experiments. The signaling pathway (Protein kinase B [Akt]/Nuclear factor erythroid 2-related factor 2 [Nrf2] pathway) of B7-H3 on the regulation of glutathione (GSH) metabolism was examined by Western blotting assay. Multi-color immunohistochemistry (mIHC) was used to detect the expression of B7-H3, cluster of differentiation 44 (CD44), and Nrf2 on human GC tissues. Student's t -test was used to compare the difference between two groups. Pearson correlation analysis was used to analyze the relationship between two molecules. The Kaplan-Meier method was used for survival analysis. RESULTS: B7-H3 knockdown suppressed the stemness of GC cells both in vitro and in vivo . Mass spectrometric analysis showed the downregulation of GSH metabolism in short hairpin B7-H3 GC cells, which was further confirmed by the experimental results. Meanwhile, stemness characteristics in B7-H3 overexpressing cells were suppressed after the inhibition of GSH metabolism. Furthermore, Western blotting suggested that B7-H3-induced activation of GSH metabolism occurred through the AKT/Nrf2 pathway, and inhibition of AKT signaling pathway could suppress not only GSH metabolism but also GC stemness. mIHC showed that B7-H3 was highly expressed in GC tissues and was positively correlated with the expression of CD44 and Nrf2. Importantly, GC patients with high expression of B7-H3, CD44, and Nrf2 had worse prognosis ( P = 0.02). CONCLUSIONS B7-H3 has a regulatory effect on GC stemness and the regulatory effect is achieved through the AKT/Nrf2/GSH pathway. Inhibiting B7-H3 expression may be a new therapeutic strategy against GC.
Humans ; Cell Line, Tumor ; Neoplasm Recurrence, Local ; NF-E2-Related Factor 2/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Signal Transduction ; Stomach Neoplasms

Background: The incidence of ulcerative colitis (UC) has gradually increased in China in recent years. The pathogenesis of UC is related to the dysfunction of immune system. B7-H5 is an important immune checkpoint molecule and is significant for the regulation of immune function. Ainis: To investigate the expression and clinical significance of B7-H5 in UC. Methods: A total of 65 UC tissue specimens were collected from Jan. 2010 to Dec. 2020 at the First Affiliated Hospital of Soochow University, and 5 healthy subjects were served as controls. Immunohistoehemistry and immunofluorescence were used to detect the expression of B7-H5, and its relationship with elinieopathologieal characteristics of UC patients was analyzed. Results: Expression of B7-H5 was significantly increased in UC patients than in controls (P < 0. 001). B7-H5 expression in UC patients was positively correlated with ESR and CRP (P < 0. 01), but not related to gender, age, extent of lesion, Mayo score and UCEIS score (P > 0. 05). Conclusions; The expression of B7-H5 in UC patients is significantly increased and is correlated with ESR and CRP, and can be used as a new marker for reflecting the severity of inflammation in UC patients.

Gastrointestinal cancers are the common malignant tumors of the digestive system, and their morbidity and mortality are in the forefront of malignant tumors. Currently, cancer immunotherapy is the hottest topic in cancer research field. Although cancer immunotherapy has achieved some results in the fundamental research and clinical application of gastrointestinal tumors, there are still a series of problems that need to be resolved. In this article, we review the fundamental and clinical research progress of several common methods of cancer immunotherapy in the field of gastrointestinal tumors.