Objective:To explore whether LBP3 exerts anti-tumor effects by promoting production of short-chain fatty acids(SCFAs)by intestinal microbiota and regulating function of polymorphonuclear myeloid-derived suppressor cells(PMN-MDSC).Methods:A subcutaneous H22 liver cancer model was employed to assess anti-tumor activity of LBP3 and its regulatory effects on PMN-MDSC.Pseudo-sterile tumor-bearing mouse model was used to investigate role of intestinal microbiota in tumor suppression of LBP3.Fecal microbiota transplantation(FMT)was conducted to explore immune regulatory role of LBP3-modulated flora.Serum SCFAs levels in tumor-bearing mice were quantified using liquid chromatography-mass spectrometry,and effect of SCFAs butyrate on arginase 1(Arg-1)expression was evaluated in vitro.Results:Both low-dose(125 mg/kg)and high-dose(250 mg/kg)LBP3 signifi-cantly inhibited tumor growth in H22 tumor-bearing mice,also led to a marked reduction in proportion of PMN-MDSC in both spleen and tumor,a reduced proportion of Treg in lymphoid tissues,a decrease in Arg-1 level within tumor,infiltration of CD8+T cells into tumor was significantly enhanced.However,these effects of LBP3 were did not observed in pseudo-sterile mice,while the above changes could be reproduced after fecal supernatant transplantation in high-dose LBP3 treatment group,suggesting a crucial role for gut microbiota.Furthermore,co-expression of Ly6G and SCFA receptor GPR43 in tumor was also observed.LBP3 treatment resulted in increased levels of SCFAs,particularly butyrate,in both blood and tumor tissues.In vitro,butyrate was shown to inhibit Arg-1 expression in MSC-2 cells,further supporting hypothesis that SCFAs mediate immune-modulatory effects of LBP3.Conclusion:LBP3 exerts its anti-tumor effects by promoting SCFA production,which subsequently inhibits function of PMN-MDSC.This highlights LBP3's potential as an immunomodulatory agent in cancer therapy.

Objective:To explore the molecular mechanisms by which miR-101-3p inhibits epithelial-mesenchymal transition(EMT)of gastric cancer cells and M2 polarization of macrophages.Methods:Bioinformatics software was used to analyze the expres-sion of miR-101-3p in gastric cancer and its correlation with survival.Transwell and Western blot were performed to evaluate the effect of miR-101-3p and Anxa2 on the migration,invasion and EMT of gastric cancer cells.Human monocytes(THP-1)were co-cultured with transfected gastric cancer cells.Immunofluorescence and Western blot assays were performed to assess the impact of miR-101-3p and Anxa2 on the expression of M2 macrophage markers CD206 and arginase-1(Arg-1).Immunohistochemistry and Western blot were used to analyze the expression of Anxa2 in gastric cancer tissues and cells.Bioinformatics software,dual-luciferase reporter and West-ern blot were utilized to validate the target relationship between miR-101-3p and Anxa2.Results:Compared with adjacent tissues,the level of miR-101-3p in gastric cancer tissues was significantly reduced and positively correlated with survival.Additionally,the levels of miR-101-3p in gastric cancer cells were significantly lower than those in normal gastric mucosal cells(P<0.01).Overexpression of miR-101-3p in gastric cancer cells significantly reduced their migration and invasion capabilities.This was accompanied by a marked decrease in the expression of N-cadherin and vimentin,and a significant increase in the expression of E-cadherin.In the co-cultured system,overexpression of miR-101-3p in gastric cancer cells significantly inhibited macrophage M2 polarization(P<0.01).Compared with adjacent tissues,the level of Anxa2 was significantly elevated in gastric cancer tissues.Compared with normal gastric mucosal cells,Anxa2 levels were significantly higher in gastric cancer cells(P<0.01).Anxa2 was target gene of miR-101-3p.Overexpression of Anxa2 in gastric cancer cells significantly enhanced their migration and invasion capabilities.In the co-culture system,overexpres-sion of Anxa2 in gastric cancer cells markedly promoted macrophage M2 polarization(P<0.01).Overexpression of Anxa2 could re-verse the inhibitory effects of miR-101-3p on EMT of gastric cancer cells and M2 polarization of macrophages(P<0.01).Conclusion:miR-101-3p regulates Anxa2 to inhibit EMT of gastric cancer cells and M2 polarization of macrophages,thereby suppressing the migra-tion and invasion of gastric cancer cells.

Objective:To investigate expression levels of various myeloid suppressor cells(MDSCs)in bone marrow of patients with chronic myeloid leukemia(CML),and the difference and correlation of expression levels of BCR-ABL fusion gene and WT1 in CML patients at different stages,and explore its clinical significance.To analyze and compare the distribution differences of various MDSCs in CML with different remission depths after treatment.Methods:Proportions of various MDSCs in 58 CML patients were de-tected by flow cytometry.Relative expressions of WT1 and BCR-ABL were detected by RQ-PCR.Iron deficiency anemia patients were served as control group,differences of the distribution of MDSCs in CML patients with different BCR-ABL expression,different WT1 expression,different CD34+cell numbers and different disease course,and expressions of various types of MDSC at 3,6,12 and 24 months after treatment in patients with chronic phase of CML were analyzed.At the same time,changes of cellular immune status in CML patients at different stages were detected,and correlations between the changes of lymphocyte subsets and MDSCs were compared.Results:Proportions of G-MDSC and e-MDSC in chronic phase of CML were significantly higher than that in normal control group(P<0.05).Proportions of G-MDSC and e-MDSC in CML patients in accelerated phase and blast crisis phase were significantly higher than that in CML patients in chronic phase(P<0.05).However,the difference in proportion of M-MDSC between accelerated phase of CML and chronic phase of CML had no statistical significance.Proportion of G-MDSC in CML patients was positively correlated with values of BCR-ABL,WT1 genes and proportion of CD34+cells(r=0.558 7,0.530 7,0.598 1),proportion of M-MDSC was positively correlated with values of BCR-ABL,WT1 genes and proportion of CD34+cells(r=0.132 1,0.144 6,0.157 8).Proportion of e-MDSC was positively correlated with values of BCR-ABL,WT1 genes and proportion of CD34+cells(r=0.604 3,0.620 7,0.625 9).G-MDSC was significantly lower in the best response group than that in warning/failure group at all stages of treatment.e-MDSC was differential in the best response and warning/failure groups at only 3 months of treatment.M-MDSC was not statistically significant in the best response and warning/failure groups at all stages of treatment.And only G-MDSC cell ratio was positively correlated with its BCR-ABL ratio(r=0.798 1).Per-centage of T lymphocyte in CML blast crisis phase was significantly lower than that in accelerated and chronic phases,while percentage of NK cells was higher.Only the proportion of G-MDSC was negatively correlated with the proportion of T lymphocyte(r=-0.815 2).Conclusion:Various MDSCs are positively correlated with BCR-ABL,WT1 gene and CD34+cells,and positively correlated with the tumor burden of CML patients,while the correlation of M-MDSC is weaker than that of G-MDSC and e-MDSC.With the remission of CML,G-MDSC decreases,while M-MDSC does not change.e-MDSC only shows differences in the early 3-month of treatment.Change of G-MDSC ratio may predict the effect of CML treatment.MDSCs can inhibit the proliferation of T lymphocyte,and inhibitory effect of G-MDSC is stronger than that of M-MDSC and e-MDSC.

Objective:To investigate the application value of combined detection of urine genes Twist1,Onecut2,VIM methyl-ation and folate metabolism related genes MTHFR(C677T/A1298C),MTRR(A66G)polymorphisms in the screening and diagnosis of bladder cancer.Methods:A total of 134 patients with primary bladder cancer admitted to the Department of Urology of Qingyang Peo-ple's Hospital and the Second Hospital of Lanzhou University from January 2023 to January 2024 were selected(bladder cancer group),and a total of 130 patients with common benign urinary system diseases and other malignant tumors of urinary system treated with cystoscopy were admitted during the same period(control group).Methylation-specific polymerase chain reaction was used to de-tect the methylation of Twist1,Onecut2 and VIM genes in urine shed cells.PCR fluorescence probe fusion was used to detect the poly-morphism of folate metabolism-related genes in peripheral blood of patients,and collected the clinical data and immunological indica-tors,and to all the data for statistical analysis.Results:The methylation rates of hematuria,bladder irritation,Twist1,Onecut2 and VIM genes were significantly different between two groups(P<0.05).The area under ROC curve(AUC)of Twist1,Onecut2 and VIM genes methylation and their combined detection were 0.721,0.675,0.674 and 0.772,respectively.Sensitivity and specificity were 73.20%and 71.00%,56.10%and 79.00%,48.80%and 86.00%,80.50%and 69.00%,respectively.The AUC of hematuria and blad-der irritation were 0.661 and 0.652.The sensitivity and specificity were 60.20%and 72.00%,41.50%and 89.00%,respectively.The combined AUC of all indicators were the largest(0.858),and the sensitivity and specificity were higher.The frequencies of CC,CT,TT,and T alleles of MTHFR C677T in bladder cancer group were 21.64%,41.79%,36.56%and 57.46%,respectively.Compared with the control group,the difference was statistically significant(P<0.05).The T allele frequency was significantly different between methylated and unmethylated Twist1 groups(P<0.05).Others differences were not statistically significant,and there was no signifi-cant association with gene methylation(P>0.05).Conclusion:The methylation of Twist1,Onecut2 and VIM genes are highly ex-pressed in the urine cells of patients with bladder cancer,and the combination of hematuria and bladder irritation has a high predictive value for the diagnosis of bladder cancer.The MTHFR(C677T)T allele is associated with the methylation of Twist1 gene and may be one of the risk factors for bladder cancer.

Objective:To investigate short-term efficacy of interventional treatment guided by abdominal superficial ultrasound and its impact on immune levels in patients with advanced pancreatic cancer.Methods:A total of 152 patients diagnosed with advanced pancreatic cancer from May 2018 to March 2020 in the People's Hospital of Yuechi were selected as study subjects,with 84 patients treated with combination chemotherapy and 68 patients treated with chemotherapy alone.After 1∶1 propensity score match-ing,a total of 60 patients were included in each group.Clinical data were compared between matched groups,and effect of different treatment methods on short-term efficacy and immune levels in advanced pancreatic cancer were evaluated by generalized estimating equations.Results:There were no statistically significant differences in age,BMI,tumor location,gender,tumor size and tumor stage between matched groups(P>0.05).When comparing efficacy of matched groups,local control rate(LCR)showed a declining trend after treatment(P<0.05),median durations of local progression-free survival(LPFS)and overall survival(OS)in chemotherapy group were higher than monotherapy group(P<0.05).There was no statistically significant difference in CA19-9 level before treatment between combination chemotherapy group and monotherapy chemotherapy group(P>0.05).In combiuation chemotherapy group,CA19-9 level showed a continuous downward trend after treatment,with a significant decrease at 6-month follow-up compared to be-fore treatment;in chemotherapy alone group,CA19-9 level showed an overall decrease followed by an increase,with the lowest point at 1 month,and a significant decrease compared to before treatment,while no significant differences in remaining follow-up periods(P>0.05).There was no significant difference of CA19-9 level at 7 days and 14 days after treatment between monotherapy chemotherapy group and combination chemotherapy group(P>0.05).However,at 1 month,3 months and 6 months after treatment,CA19-9 level in combination chemotherapy group was significantly lower than monotherapy chemotherapy group(P<0.05).Immune levels in combina-tion chemotherapy group showed an initial increase followed by a decrease,reaching the highest point at 1 month,and total T-cell count showed a significant decrease after 6 months of treatment compared to before treatment(P<0.05).CD3+T,NK,IL-2 and IFN-γ immune levels were still higher than before treatment(P<0.05),with no significant differences in remaining indicators compared to before treatment(P>0.05).CD4+T/CD8+T in chemotherapy alone group did not show a significant trend,and remaining indicators showed same trend as combined chemotherapy group(P<0.05);after 6 months of treatment,total T-cell count,CD3+T cell count,NK cell count and IL-2 level were lower than before treatment(P<0.05),with no significant differences in remaining indicators compared to before treatment(P>0.05).During follow-up period,combined chemotherapy group showed higher levels of total T cells,CD3+T cells,NK cells,IL-2,IFN-γ and IL-6 levels compared to single chemotherapy group(P>0.05).CD4+T/CD8+T at 14 days and 3 months of follow-up was significantly higher in combined chemotherapy group than single chemotherapy group(P<0.05),while differences were not significant at other follow-up times(P>0.05).Generalized estimating equation evaluation showed that all indica-tors were statistically significant.Conclusion:Interventional treatment guided by abdominal superficial ultrasound can effectively improve short-term efficacy and immune levels in patients with advanced pancreatic cancer.

Objective:To explore the potential value of dynamic changes of inflammatory factors in evaluating the efficacy of platinum regimen in the first-line treatment of advanced lung adenocarcinoma(LUAD).Methods:A total of 121 patients with advanced LUAD without common target mutations who were admitted to Dongtai People's Hospital to receive first-line treatment with platinum regimen from January 2021 to January 2023 were selected,and relevant clinical data such as general information,hematuria routine,angiogenic factors,serum tumor markers,inflammatory factors,immunoglobulin and T lymphocyte indexes were collected and statisti-cally analyzed.Patients in complete response(CR)group and partial response(PR)group were classified as chemotherapy sensitive group;stable(SD)group and progressive(PD)group were classified as chemotherapy insensitive group according to response evaluation criteria in solid tumors(RECIST).Through univariate and multi-factor Logistics regression analysis,the influencing factors of first-line treatment efficacy of platinum regimen in patients with advanced LUAD were analyzed,and the regression equation y=1-1/(1+e-z)prediction model was established and verified.Receiver operating characteristic(ROC)curve was used to analyze the potential value of dynamic changes of inflammatory factors in evaluating the efficacy of platinum-based first-line treatment for advanced LUAD.Results:①Univariate and multivariate Logistic regression analysis showed that CEA,TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were still the influencing factors for the efficacy of first-line chemotherapy of platinum regimen in patients with advanced LUAD(P<0.05).②Inflammatory factors TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were included in multivariate Logistic regression analysis.After correcting confounders(Model 5),high levels of TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were positively correlated with insensitivity after first-line chemotherapy of platinum regimen in advanced LUAD patients(P<0.05).Restrictive cubic spline model analysis showed that the dynamic changes of inflammatory factors and the insensitivity of patients with advanced LUAD after first-line chemotherapy all had a nonlinear dose-response relationship.With the increases of TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP levels,patients with advanced LUAD have a greatly increased risk of desensitization after first-line chemotherapy.③Stratified interaction test analysis showed that tumor stage,differentiation degree and lymph node metastasis before and after correction factors were significantly correlated with expressions of inflammatory factors(all P<0.05,all P interaction<0.05).④Prediction model based on the influence factors of inflammatory factors was well distinguished and accurate,the area under ROC curve(AUC)before and after internal validation for detecting chemotherapy sensitivity in patients with advanced LUAD treated with platinum regimen first-line therapy was 0.87(95%CI:0.81～0.93)and 0.88(95%CI:0.82～0.95),respectively,and the sensitivity were 89.69%and 89.75%,respectively,the specificity were 91.77%and 91.85%,respectively.Conclusion:Dynamic changes of inflammatory factors is an important factor affecting the chemotherapy efficacy of patients with advanced LUAD treated with platinum-containing regimen.With the significant decrease of inflammatory factors,the sensitivity of patients to chemotherapy is also significantly increased.

Objective:To investigate the effect of TCF1+CD8+T cells on the prognosis of nasopharyngeal carcinoma(NPC)pa-tients undergoing immunotherapy.Methods:A retrospective study was conducted on 108 NPC patients admitted to Anyang Cancer Hospital from January 2018 to April 2020.Pathological findings and clinical data were collected,and multiple immunofluorescence staining was utilized to detect TCF1+CD8+T cell levels in tumor tissue.Cell counts and the proportion of positive cells were measured,and the optimal cut-off value was determined using a time-dependent receiver operating characteristic(ROC)curve.Patients were clas-sified into high expression group and low expression group according to the cut-off value.The general information and survival status were compared between two groups,and univariate and multivariate Cox regression analysis were performed to screen the influencing factors of patient prognosis.Results:The optimal cut-off value was determined using ROC curve.A cut-off value of 0.34%for TCF1+CD8+T cell area under the curve(AUC)=0.653 resulted in a sensitivity of 73.5%and a specificity of 43.7%.Based on the opti-mal cut-off value,33 patients with TCF1+CD8+T cell beyond 0.34%were included in high expression group,and the remaining 75 pa-tients were included in low expression group.The proportion of TCF1+CD8+T cells was(0.43±0.09)%in high expression group and(0.21±0.08)%in low expression group.Comparison of general data yielded that the gender,age,body mass index(BMI),clinical stage,and pathological classification showed no statistical difference between two groups(P>0.05),while the percentage of patients with T stage of T3 and T4,N stage of N2 and N3,and M stage of M1 in high expression group were smaller than that in low expression group(P<0.05).The 24-month survival rate was 81.82%(27/33)in high expression group and 74.67%(56/75)in low expression group,with no statistical difference between two groups(χ2=0.658 8,P=0.417 0).Univariate and multivariate Cox regression analysis found that T stage,N stage and M stage were independent risk factors affecting the prognosis of patients,and TCF1+CD8+T expression was a protective factor.ROC curve indicated that the AUC,sensitivity and specificity of TCF1+CD8+T expression in predicting death of NPC patients undergoing immunotherapy was 0.674,69.7%and 65.3%,while the AUC,sensitivity and specificity of TCF1+CD8+T ex-pression combined with TNM stage in predicting death was 0.809,75.8%and 84.0%,respectively.Conclusion:NPC patients with higher proportion of TCF1+CD8+T cells and lower TNM stage have higher 24-month survival rate after immunotherapy,and the above parameters are of great predictive value for patient prognosis.

Vast majority of human RNA are non-coding RNA(ncRNA),which can regulate physiological activities and affects functions on cells.Although each part of them is obscure,they are involved in different levels of gene regulation,from epigene-tic gene silencing to post transcriptional regulation.Metabolic reprogramming of tumor-associated macrophages(TAMs)can promote tumor progression and is closely related to activation phenotypes of anti-tumor(M1)or pro-tumor(M2).A lot of ncRNAs are involved in metabolic reprogramming of TAMs,reshaping tumor microenvironment and playing a key role in tumor progression.Improving understanding of effect on ncRNA regulating metabolic reprogramming of TAMs may provide new methods for developing effective clinical treatments.

Tumor growth and metastasis is a multistep process dependent on the host microenvironment.While the immune system can suppress cancer development,it can also promote tumor growth and progression through a variety of mechanisms.In this complex network of immune regulatory systems,the establishment and maintenance of immune evasion mechanisms are critical for can-cer progression.Recent studies have found that erythroid precursor cells(EPCs)play an important role in regulating immune responses and tumor progression.EPCs are immature erythroid cells that differentiate into mature erythrocytes under physiological conditions,but tumor-induced EPCs are stagnant in differentiation,leading to expansion in extramedullary hematopoietic sites such as the liver and spleen,negatively regulating immune function,and promoting tumor growth and metastasis.This paper summarizes the mecha-nism of EPCs suppressing immunity,their role in promoting tumor cell immune escape,and the mechanism of promoting tumor pro-gression,which provides a new idea for the future study of the immune mechanism of liver metastases and is conducive to finding fu-ture effective therapeutic target for liver metastases.

Tumor-targeted therapy is a vital approach in precision medicine,selection of target molecule is a key element in tumor-targeted therapy.B7-H3(CD276),as an immune checkpoint molecule selectively highly expressed on tumor cells,is a crucial molecule involved in regulation of tumor immunity.Numerous recent studies have found that B7-H3,in addition to its immune check-point function,also participates in regulation of malignant biological behaviors of tumor cells and is considered to have great potential as a pan-cancer universal drug target.Current immunotherapeutic strategies targeting B7-H3 primarily include development of block-ing monoclonal antibodies(Blocking mAb),antibody-drug conjugate(ADC),chimeric antigen receptor(CAR)T/NK cell therapies,monoclonal antibody dependent cellular cytotoxicity(ADCC)drugs,radionuclide drug conjugates(RDC),bispecific/trispecific anti-body(BsAb/TriAb),and antibody cytokine drug combinations,among others.These strategies aim to enhance body's immune re-sponse to cancer through various mechanisms,thereby inhibiting tumor growth and metastasis.Multiple preclinical and clinical studies have shown that immunotherapy targeting B7-H3 has potential therapeutic value.Therefore,this article mainly reviews the latest status of development of tumor immunotherapy drugs targeting B7-H3 molecule.