Abstract:Objective To explore the value of CD8⁺T cell infiltration in osteosarcoma biopsy tissues before treatment in predicting the response to neoadjuvant chemotherapy(NACT)and the prognosis of patients.Methods A total of 185 patients with limb osteosarcoma admitted to Beijing Jishuitan Hospital,Capital Medical University from January 1,2021 to April 23,2025 were selected and divided into a training cohort(n=129)and a validation cohort(n=56).Biopsy tumor tissue samples of the patients before treatment were collected.The infiltration density of CD8⁺ T cells in tumor tissues was detected by immunohistochemical staining,and the patients were divided into the high infiltration group and the low infiltration group based on the median value of the infiltration density.The relationship between CD8⁺ T cell infiltration and the therapeutic effect of NACT was analyzed by the Logistic regression model.The overall survival time(OS),metastasis-free survival time(DMFS),and tumor-free survival time(DFS)of the patients were followed up.The survival curve was plotted using the Kaplan-Meier method,and the survival difference was tested by Log-rank.Results The density of CD8⁺ T cells in osteosarcoma biopsy tissues of the training cohort ranged from 0.74 to 1 322.10 cells/mm²,with a median value of 25.28 cells/mm².The verification cohort ranged from 0.70 to 548.71 pieces/mm²,with a median of 18.79 pieces/mm².Both the training cohort and the validation cohort showed that high infiltration of CD8⁺ T cells was significantly correlated with NACT sensitivity(P<0.001).The results of multivariate Logistic regression analysis showed that CD8⁺ T cell infiltration was a significant factor in predicting the efficacy of NACT(OR=0.03,P<0.001).The DMFS and DFS of patients in the high infiltration group were significantly better than those in the low infiltration group.Conclusion The infiltration level of CD8⁺ T cells in osteosarcoma tissues before treatment is closely related to the therapeutic effect of NACT.Patients with high infiltration benefit more from NACT.The infiltration level of CD8⁺ T cells can serve as a potential biomarker for predicting the efficacy of NACT.

Abstract:Objective To investigate the predictive value of PET/CT metabolic heterogeneity parameters in primary tumors and metastatic lymph nodes for chemoradiation sensitivity and survival prognosis in stage Ⅲ non-small cell lung cancer(NSCLC).Methods This was a prospective cohort study enrolling 120 unresectable stage Ⅲ NSCLC patients from October 2020-December 2021.Patients were divided into a high metabolic heterogeneity group(SUVmax difference>25%,n=54)and a low metabolic heterogeneity group(SUVmax difference≤25%,n=66)based on the maximum standard uptake value(SUVmax)difference between primary tumors and metastatic lymph nodes from pretreatment PET/CT images.Metabolic heterogeneity parameters(including SUVmax,entropy,skewness,kurtosis,etc.),short-term efficacy[objective response rate(ORR),disease control rate(DCR)],toxicity reactions were analyzed,and Kaplan-Meier curves were plotted to compare progression-free survival(PFS),and the correlation between RECISTv1.1 and PET/CT metabolic assessment results was verified.Results Before treatment,the primary tumor SUVmax(8.78±2.32 vs.6.57±1.89),SUVmax difference(3.56±1.25 vs.0.82±0.51),SUVmax difference percentage[(38.53±12.37)%vs.(10.22±5.77)%],entropy(1.75±0.38 vs.1.12±0.25),skewness(0.82±0.45 vs.0.35±0.28),and kurtosis(3.15±1.20 vs.2.08±0.95)in the high metabolic heterogeneity group were significantly higher than those in the low metabolic heterogeneity group,while the lymph node SUVmax(5.23±1.56 vs.6.35±1.97)was significantly lower than that in the low metabolic heterogeneity group(all P<0.001).After treatment,the primary tumor SUVmax(4.21±1.53 vs.3.05±1.21),lymph node SUVmax(4.87±1.12 vs.3.26±1.03),SUVmax difference(0.45±0.31 vs.0.12±0.08),and SUVmax difference percentage[(9.82±5.63)%vs.(3.56±2.17)%]in the high metabolic heterogeneity group were still significantly higher than those in the low metabolic heterogeneity group(all P<0.001).The ORR(65.15%vs.40.74%,P=0.008)and DCR(89.39%vs.72.22%,P=0.016)in the low metabolic heterogeneity group were significantly higher than those in the high metabolic heterogeneity group.There was no significant difference in the incidence of acute and late toxic reactions between the two groups(P>0.05).The overall consistency rate between RECISTv1.1 and PET/CT metabolic assessment results was 87.50%,with a Kappa value of 0.78(P<0.001).Conclusion Metabolic heterogeneity between primary tumors and metastatic lymph nodes in stage Ⅲ NSCLC is closely associated with chemoradiation sensitivity and prognosis.SUVmax difference percentage may serve as a potential imaging indicator for predicting chemoradiation efficacy and progression-free survival,providing evidence for personalized treatment.

Abstract:Objective To explore the short-term efficacy and long-term prognosis of transcatheter arterial chemoembolization(TACE)after repeated hepatectomy/ablation in patients with recurrent early hepatocellular carcinoma(HCC).Methods A retrospective analysis was conducted on 178 HCC patients who underwent repeated radical hepatectomy or ablation due to early tumor recurrence(within 2 years after surgery)at Jilin Provincial Cancer Hospital from January 2020 to May 2023.According to whether they received adjuvant TACE treatment,patients with recurrent HCC were divided into the TACE group(adjuvant TACE after repeated liver resection or ablation,n=89)and the control group(close monitoring after repeated liver resection or ablation,n=89).Record the adverse reactions of the patients and follow up on recurrence-free survival(RFS).Results The median follow-up time for all patients in the group was 23.0(4.0-60.0)months,the cumulative recurrence rate was 71.91%(128/178),and 75 cases died.By the end of the follow-up,the recurrence-free survival in the TACE group and the control group was 29 cases and 21 cases,respectively.The median RFS time in the TACE group was longer than that in the control group(28 months vs.16 months),but the difference was not statistically significant(Log-rank χ 2=2.897,P=0.084).The most common adverse reaction related to TACE was fever(40 cases,44.94%).Other adverse events include elevated liver enzymes,abdominal pain,elevated bilirubin,nausea/vomiting,and leukopenia.All adverse reactions were grade 3 or below,and there were no deaths related to TACE.The results of multivariate Cox regression analysis showed that radiofrequency ablation(RFA),multicenter recurrent lesions in the liver,Child-Pugh classification,liver cirrhosis and microvascular invasion were significant predictive factors affecting RFS(P<0.05).The survival risk score was calculated based on the Cox regression coefficients corresponding to the four indicators of RFA,multicenter recurrent lesions in the liver,Child-Pugh classification and microvascular invasion.Stratification was based on the cut-off threshold of survival risk score(-0.29):89 cases in the high-risk group and 89 cases in the low-risk group.In the high-risk group,compared with the control group,the median RFS time of patients in the TACE group was longer(17.0 months vs.13.0 months,Log-rank χ 2=5.367,P=0.021).Conclusion Adjuvant TACE after repeated radical therapy in patients with early recurrent HCC has no significant association with RFS in the overall cohort.However,it may be beneficial for specific subgroups with high-risk factors for recurrence,including patients receiving second-line RFA,those with multicenter intrahepatic recurrence foci,Child-Pugh grade B,and those with microvascular invasion.

Abstract:Poly(ADP-ribose)polymerase inhibitors(PARPi)have inaugurated a new era of synthetic-lethal therapy for ovarian cancer;however,almost 50%of patients develop primary or acquired resistance during first-line or maintenance treatment,underscoring the need for additional synergistic targets.Ferroptosis—an iron-dependent,lipid-peroxidation-driven form of programmed cell death—has emerged as a promising complement.Recent studies show that PARPi weaken tumor antioxidant defenses and heighten oxidative stress,thereby lowering the ferroptotic threshold.In BRCA-wild-type or homologous-recombination-proficient ovarian-cancer models,combining PARPi with ferroptosis inducers markedly restores drug sensitivity and suppresses tumor growth.This review summarizes the molecular crosstalk between PARP inhibition and ferroptosis,highlights representative combination strategies,and discusses translational challenges,aiming to chart new avenues for overcoming PARPi resistance.

Abstract:Objective To explore the application effect of labial suture combined with C-type tunnel extraperitoneal fistula in laparoscopic Miles surgery for rectal cancer.Methods A total of 200 patients with low rectal cancer who underwent laparoscopic Miles surgery for rectal cancer from March 2023 to December 2024 were collected and divided into a control group(surgical method of tubular stapler+intraperitoneal fistula)and an observation group(surgical method of labial suture+C-type tunnel extraperitoneal fistula),with 100 cases in each group.The surgery-related indicators,occurrence of complications,recurrence and death of the two groups of patients were compared.Results The time of the first bowel sound in the observation group was(2.02±0.21)days,the time of the first flatus was(3.04±0.27)days,the time of the first defecation was(4.12±0.21)days,and the length of hospital stay was(11.25±1.14)days.The levels of patients in the control group were(2.87±0.43)d,(3.95±0.59)d,(4.98±0.57)d and(14.28±2.05)d respectively.There were statistically significant differences between the two groups(all P<0.05).The incidence of complications in the observation group at 3 months and 6 months after the operation was 5.00%(5/100)and 3.00%(3/100)respectively,which was lower than that in the control group at the same period[22.00%(22/100),29.00%(29/100)](all P<0.05).No recurrence or death occurred in either group of patients.Conclusion Labial suture combined with C-tunnel extraperitoneal fistula can effectively improve perioperative related indicators in patients with low rectal cancer,accelerate the recovery of intestinal function after surgery,and reduce the incidence of complications.

Abstract:Objective To study the expression of apolipoprotein D(APOD)in cervical squamous cell carcinoma and the regulatory mechanism of APOD expression level on the proliferation of cervical squamous cell carcinoma cells.Method The expression of APOD in normal cervical tissues and cervical squamous cell carcinoma tissues was analyzed using GSE39001 data.The relationship between the expression level of APOD and the tumor proliferation-related gene set and the prognosis of patients was analyzed using the TCGA database.The APOD overexpression plasmid was transfected into the SiHa cell line and SW756 cell line of cervical squamous cell carcinoma.The effects of APOD overexpression on cell proliferation ability and cell cycle were detected.Western blotting was used to detect the expression of related proteins in the PTEN/Akt signaling pathway.Results GSE39001 dataset showed that the expression of APOD in the tissues of patients with cervical squamous cell carcinoma was significantly decreased compared with normal cervical tissues(P<0.001).TCGA data analysis shows that the low expression of APOD in cervical squamous cell carcinoma tumor tissues is significantly negatively correlated with the tumor proliferation-related gene set.Kaplan-Meier survival curve analysis showed that the prognosis of patients with cervical squamous cell carcinoma in the APOD low expression group was relatively poor(HR=0.543,95%CI:0.328-0.9,P<0.05).Compared with the SiHa-Control group and the SW756-Control group,the absorbance values(A values)of cells in the SiHa-APOD-OE group and the SW756-APOD-OE group were significantly decreased(P<0.05).Compared with the SiHa-Control group,the proportion of cells in the G1 phase in the SiHa-APOD-OE group increased significantly[(15.45±2.78)%vs.(28.34±4.01)%].The proportion of cells in the S phase decreased significantly[(48.32±4.35)%vs.(33.05±4.28)%],and the differences were statistically significant(P<0.05).Compared with the SiHa-Control group and the SW756-Control group,the expression of PTEN protein in the SiHa-APOD-OE group and the SW756-APOD-OE group was significantly increased(P<0.05),and the expression of p-Akt/Akt was decreased(P<0.05).Conclusion APOD is significantly lowly expressed in cervical squamous cell carcinoma tissues and negatively regulates the PTEN/Akt signaling pathway,thereby affecting the cell cycle and inhibiting tumor cell proliferation.

Abstract:Objective To explore the clinical efficacy and safety of modified radical mastectomy and open surgery for breast cancer under laparoscopy.Methods A retrospective analysis was conducted on a total of 160 female patients who underwent modified radical mastectomy for breast cancer in the Affiliated Hospital of North Sichuan Medical College from January 2021 to January 2025.They were divided into the endoscopic group(n=60)and the open group(n=100)according to the surgical methods.Compare the differences between the two groups of patients in terms of perioperative indicators,postoperative complications,postoperative quality of life and clinical efficacy.Results There were no statistically significant differences in baseline characteristics such as age,gender,BMI,tumor size,number of sentinel lymph nodes,TNM stage and menstrual status between the two groups of patients(P>0.05).The operation time[(67.2±11.7)min vs.(92.1±10.6)min],intraoperative blood loss[(57.8±11.5)mL vs.(40.1±9.6)mL],and drainage volume[(1 605.7±275.6)mL vs.[(865.7±131.6)mL],drainage time[(8.7±2.9)d vs.(7.0±1.9)d],incision length[(21.4±3.9)cm vs.(8.6±1.5)cm]and postoperative hospital stay[(4.1±1.2)d vs.(3.5±1.1)d]were statistically significant(P<0.05).The total incidence of postoperative complications in the endoscopic group was significantly lower than that in the open group(6.7%vs.19.0%,P<0.05).The patient was followed up for 6 months after the operation In the endoscopic group,patients had pain scores on the affected upper limb[(6.5±1.5)points vs.(8.7±1.4)points],chest wall pain scores[(7.0±1.7)points vs.(8.9±0.9)points],and shoulder joint range of motion scores[(7.6±1.2)points vs.(8.7±1.3)points]and arm activity function score[(7.7±1.4)points vs.(8.9±0.9)points]were significantly better than those in the open group(P<0.05).At the end of the follow-up,there was no statistically significant difference between the two groups of patients in clinical efficacy indicators such as survival rate,local recurrence rate and distant metastasis rate.Conclusion Compared with open surgery,modified radical mastectomy for breast cancer by laparoscopy has significant advantages in perioperative indicators,postoperative complications and postoperative quality of life,and the clinical efficacy is comparable.

Abstract:Objective To explore the relationship between low expression of human epidermal growth factor receptor 2(HER-2)and the clinicopathological characteristics and prognosis of breast cancer.Methods A retrospective analysis was conducted on the clinical medical records of patients with HER-2 negative breast cancer who were treated at the Breast Disease Center of Shaoyang Central Hospital from 2014 to 2019.The expression of HER-2 was detected by immunohistochemistry(IHC)and fluorescence in situ hybridization(FISH),and the patients were divided into those with low HER-2 expression(IHC 1+and IHC 2+without amplification by FISH)and those with zero HER-2 expression(IHC 0).Positive hormone receptor(HR)is defined as estrogen receptor(ER)status and/or positive progesterone receptor(PR).Compare the differences in clinicopathological characteristics and prognosis[disease-free survival(DFS)and overall survival(OS)]between the HER-2 low expression group and the zero expression group.Results In the total population,there were significant differences between the HER-2 low expression group and the HER-2 zero expression group in terms of tumor T stage,N stage,histological grade,Ki-67 and HR status(P<0.05).In the total population and the HR-positive group,the HER-2 low expression group was associated with a higher T stage(P<0.05);However,in the HR-negative group,low expression of HER-2 was associated with a lower T stage(P<0.05).Whether in the total population or the population grouped by HR,HER-2 zero expression was associated with a higher N stage(P<0.05).In the total population and the HR-negative group,the proportion of patients with high histological grade and high Ki-67 was higher in the HER-2 zero expression group(P<0.05).In addition,the proportion of HR positive patients in the HER-2 low expression group was significantly higher(83.33%vs.69.26%,P<0.001).Survival curve analysis showed that low expression of HER-2 was associated with poorer prognosis of DFS and OS in HR-positive patients(P<0.05).Univariate and multivariate Cox regression analyses showed that low HER-2 expression was an independent influencing factor for the prognosis of DFS(HR:3.645,95%CI:1.406-9.446,P=0.019)and OS(HR:3.512,95%CI:1.288-9.575,P=0.033)in HR-positive patients.Conclusion Among HR-positive patients,low expression of HER-2 is associated with a higher tumor T stage and a poorer survival prognosis.

Abstract:Objective To explore the diagnostic value of ultrasound elastography(UE)parameters combined with serum matrix metalloproteinase-9(MMP-9),vascular endothelial growth factor(VEGF),and soluble interleukin-2 receptor(sIL-2R)for lymph node metastasis(LNM)in thyroid cancer.Methods A total of 208 patients with thyroid cancer admitted to the First Affiliated Hospital of Henan University of Science and Technology from January 2021 to October 2024 were selected as the study group.According to whether the selected patients had lymph node metastasis,they were divided into the metastasis group(83 cases)and the non-metastasis group(125 cases).In addition,208 patients with benign thyroid tumors admitted during the same period were selected as the control group in 1:1 ratio.The clinical data,UE parameters,and levels of serum MMP-9,VEGF,sIL-2R between the study group and the control group were compared,the clinical data,UE parameters,and levels of serum MMP-9,VEGF,sIL-2R between the metastatic and non-metastatic groups were compared.The risk factors for LNM in thyroid cancer were analyzed by multivariate Logistic regression analysis,and the diagnostic value of UE parameters and serum MMP-9,VEGF,and sIL-2R combined examination for LNM in thyroid cancer were analyzed by receiver operating characteristic(ROC)curves.Results The blue area ratio,elasticity ratio,and levels of serum MMP-9 and VEGF in the study group were higher than those in the control group,while the level of serum sIL-2R was lower than that in the control group(P<0.05).The proportion of patients with multifocal and bilateral tumors in the metastasis group were 40.96%and 54.22%,respectively,which was higher than the 13.60%and 24.80%in the non-metastasis group(P<0.05).The blue area ratio,elasticity ratio,and levels of serum MMP-9 and VEGF in the metastatic group were higher than those in the non-metastatic group,while the level of serum sIL-2R was lower than that in the non-metastatic group(P<0.05).The results of multiple Logistic regression analysis showed that multifocal tumors(OR=2.838),bilateral tumors(OR=2.406),high blue area ratio(OR=2.479),high elasticity ratio(OR=3.050),high levels of serum MMP-9(OR=1.779),high levels of serum VEGF(OR=2.447),and low levels of serum sIL-2R(OR=2.155)were significant risk factors for LNM in thyroid cancer(P<0.05).Classifying LNM as positive and non-metastatic as negative,the area under cure(AUC)of blue area ratio,elasticity ratio,MMP-9,VEGF,sIL-2R,and combined examination for diagnosing LNM in thyroid cancer were 0.735,0.685,0.703,0.715,0.680,and 0.918,respectively.The sensitivity were 68.67%,63.86%,63.86%,62.65%,67.47%,and 87.95%,and the specificity were 70.40%,71.20%,69.60%,72.00%,65.60%,and 86.40%,respectively.Among them,the AUC of combined examination was the highest(P<0.05).Conclusion The parameters of UE,along with serum levels of MMP-9,VEGF,and sIL-2R,are not only closely associated with the occurrence of thyroid cancer but also serve as risk factors for LNM.The combined detection of these indicators had high diagnostic value for LNM in thyroid cancer.

Abstract:Objective To investigate the effects of sodium aescinate(SA)on the proliferation and migration of nasopharyngeal carcinoma cells,as well as its regulatory mechanism on Kelch-like epichlorohydrin-related protein-1(Keap1)/nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods CNE1 cells were treated with low and high doses of SA,the functional salvage experiment was carried out with cobalt protoporphyrin(CoPP),a HO-1 agonist.Functional recovery test with ferroptosis inhibitor Ferrostatin-1(Fer-1).3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromid(MTT)was used to detect the growth activity.Cell proliferation and apoptosis were assessed using the plate clone formation and AO/EB double staining,respectively.Transwell chamber was used to detect the cell migration.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of epithelial mesenchymal transition(EMT)related genes.Western blotting was used to detect the expression levels of Keap1,Nrf2,HO-1,Gpx4,SLC7A11 and TFR1 in cells.Results SA significantly inhibited the growth,migration,and proliferation of nasopharyngeal carcinoma CNE1 cells(P<0.05).AO/EB double staining results showed that SA markedly promoted the apoptosis of CNE1 cells(P<0.05).RT-qPCR analysis revealed that SA significantly downregulated the mRNA expression of N-cadherin and Vimentin while upregulating E-cadherin mRNA expression(all P<0.05).Western blotting results demonstrated that SA notably decreased the protein expression of Nrf2,HO-1,Gpx4,and SLC7A11,and increased the expression of Keap1 and TFR1(all P<0.05).Functional rescue experiments indicated that CoPP could partially reverse the antitumor effects of SA(P<0.05).Functional recovery experiments showed that Fer-1 could partially restore the inhibitory effects of SA on cancer cell proliferation and migration(P<0.05).Conclusion SA effectively suppressed the EMT process,and reduced the proliferation and migration abilities of nasopharyngeal carcinoma CNE1 cells,which might be associated with its regulation of the Keap1/Nrf2/HO-1 signaling pathway.

Abstract:Objective To investigate the relationship between immune cell infiltration density in tumor parenchyma and stroma and postoperative prognosis of gastric cancer.Methods Forty-seven eligible samples were screened from the gastric cancer omics cohort of the Affiliated Hospital of Jiangsu University from January 1,2016 to December 31,2018.Multiple immunofluorescence staining was performed to detect immune cells,including CD8+T cells,M1 and M2 macrophages,natural killer(NK)cell CD56bright and CD56dim subgroups.The infiltrated cell density was calculated as the number of cells per square millimeter(cells/mm2)in tumor parenchyma and stroma,respectively.The relationship of cell density with overall survival(OS)of patients after surgery was analyzed.The infiltrated cell density along with clinical pathological features were incorporated into the least absolute shrinkage and selection operator(LASSO)Cox regression model.The optimal variables for prognostic model were selected to calculate the risk score.Results The median density(number of cells/mm2)of various immune infiltrating cells in tumor tissue was as follows:CD8+T cells 75,M1 macrophage 35,M2 macrophage 118,CD56bright NK cells 64;CD56dim NK cells 670.The median density(number of cells/mm2)of various immune infiltrating cells in tumor stroma was as follows:CD8+T cells 152,M1 macrophage 96,M2 macrophage 109,CD56bright NK cells 12,CD56dim NK cells 30.The density of 5 types of cells were divided into high(≥median)and low(<median)groups based on their median values.In the tumor parenchyma,the proportion of CD8+T cells with a high density in males was significantly lower than that in females(P=0.039),the proportion of M1 macrophages with a high density in poorly differentiated tumors was significantly lower than that in well/moderately differentiated tumors(P=0.039),and the proportion of M2 macrophages with a high density in stage Ⅲ tumors was significantly higher than that in stage Ⅰ/Ⅱ tumors(P=0.018).In the tumor stroma,the proportion of M1 macrophages with a high-density in poorly differentiated tumors was higher than that in moderately differentiated tumors(P=0.039).In the tumor parenchyma,high-density CD56dim NK cell infiltration significantly prolonged OS compared to low-density individuals(P=0.001),with a median OS of not yet reached(NR)compared to 22.0 months(95%CI:7.62-36.38).The OS of high-density macrophage M1 infiltration was significantly prolonged compared to low-density individuals(P=0.007),with a median OS of NR compared to 21.0 months(95%CI:8.48-33.52).In the tumor stroma,the OS of high-density macrophage M1 infiltration was significantly prolonged compared to low-density individuals(P=0.005),with a median OS of NR compared to 21.0 months(95%CI:14.74-27.26).The postoperative pathological staging,CD56dim NK cell infiltration degree in tumor parenchyma,M1 macrophage infiltration degree in tumor parenchyma,and M1 macrophage infiltration degree in tumor stroma were screened by the LASSO regression as variables for the prognostic model,and risk scores were calculated.According to the median risk score,patients were divided into high-risk(≥median)and low-risk(<median)groups.The OS of the high-risk group was significantly lower than that of the low-risk group(P<0.001),with a median OS of 21.0 months(95%CI:11.40-30.60)compared to NR.Conclusion The infiltration density of immune cells in tumor parenchyma and stroma is closely associated with postoperative OS of gastric cancer and can serve as prognostic biomarkers.

Abstract:Objective To investigate the effects of plumbagin(PLB)on the growth,invasion,and apoptosis of gastric cancer cells by regulating the expression of hypoxia inducible factor-1α(HIF-1α)and programmed death ligand-1(PD-L1).Methods Gastric cancer cells MGC-803 were assigned into control group(conventional culture),PLB group(8 μmol/L PLB),PLB+pcDNA-NC group and PLB+pcDNA-HIF-1α group,respectively.Cell proliferation,invasion and apoptosis were detected by MTT assay,plate clone formation experiments,Transwell experiment and Flow cytometry,respectively.The mRNA and protein expressions of HIF-1α and PD-L1 were detected by quantitative real time PCR(RT-qPCR)and Western blotting,respectively.Transplant tumor models were constructed based on the MGC-803 cell lines.The twenty-four mice were randomized into four groups(n=6):a control group given intraperitoneal saline,and PLB,PLB+pcDNA-NC,and PLB+pcDNA-HIF-1α groups all treated with daily intraperitoneal injections of 6 mg/kg PLB for 6 weeks.Tumor volume and weight were measured,and the mRNA and protein expression levels of HIF-1α and PD-L1 in tumor tissues were detected by qRT-PCR and Western blotting experiments.Results Compared with the control group,the cell survival rate,colony formation rate,number of invasive cells,HIF-1α and PD-L1 mRNA and protein expression levels of gastric cancer cells in the PLB group were lower,and the apoptosis rate was higher,the volume and weight of transplanted tumors and the expression of HIF-1α and PD-L1 mRNA and protein in tumor tissues were also decreased(P<0.05).Compared with the PLB+pcDNA-NC group,the cell survival rate,colony formation rate,number of invasive cells,HIF-1α and PD-L1 mRNA and protein expression levels of gastric cancer cells in the PLB+pcDNA-HIF-1α group were higher,while the cell apoptosis rate was lower,the volume and weight of transplanted tumors and the expression of HIF-1α and PD-L1 mRNA and protein in tumor tissues were also increased(P<0.05).Conclusion PLB may downregulate PD-L1 by inhibiting HIF-1α,thereby inhibiting gastric cancer cell growth and invasion,and promoting cell apoptosis.

Abstract:Objective To investigate the expression changes of F-box only protein 21(FBXO21)in kidney clear cell carcinoma(KIRC),evaluate its prognostic significance,and preliminarily explore its potential biological mechanisms.Methods Public datasets from The Cancer Genome Atlas(TCGA)and International Cancer Genome Consortium(ICGC)were used to analyze the expression of FBXO21 in KIRC and its correlation with clinicopathological features and patient prognosis.Real-time fluorescence quantitative PCR(RT-qPCR)and immunohistochemistry(IHC)were performed on KIRC clinical samples from Meizhou People's Hospital to validate the expression patterns.Kaplan-Meier survival analysis and Cox proportional hazards regression model were used to assess its independent prognostic value,and a nomogram model was constructed.Gene Set Enrichment Analysis(GSEA),Gene Ontology(GO),and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were used to explore related biological pathways.The ESTIMATE and TIMER algorithms were applied to evaluate the relationship between FBXO21 and the tumor immune microenvironment.Results FBXO21 was significantly downregulated in KIRC tissues and its low expression was closely associated with T stage,clinical stage and histological grade(P<0.05).The RT-qPCR and IHC results were consistent with findings from public datasets(P<0.001).Survival analysis indicated that patients with low FBXO21 expression had significantly shorter overall survival and disease-free survival(P<0.001).Multivariate Cox regression confirmed FBXO21 as an significant prognostic factor(P<0.05).The nomogram model demonstrated good predictive performance.GSEA showed that FBXO21-related genes were enriched in transforming growth factor-β(TGF-β)signaling,KRAS signaling,leukin-6/Janus kinase/signal transducer and activator of transcription 3(IL6-JAK-STAT3)signaling pathway,forkhead box O(FoxO)transcription factor signaling,and other key pathways.Immune analysis revealed that FBXO21 expression was closely associated with immune scores and the infiltration of multiple immune cells.Conclusion FBXO21 is lowly expressed in KIRC and demonstrates favorable prognostic value,suggesting its potential as a biomarker and therapeutic target.

Abstract:Objective To explore the correlation between the expression of transcription factor CP2-like 1(TFCP2L1),dual-specificity phosphatase 26(DUSP26),and mixed lineage kinase domain-like(MLKL)proteins in differentiated thyroid cancer and the clinical pathological characteristics as well as the therapeutic efficacy of iodine-131(131I)treatment.Methods A total of 131 patients with differentiated thyroid cancer who underwent radical surgery and received their first 131I treatment from March 2021 to March 2024 in Qinhuangdao Hospital of Dongfang Hospital were prospectively selected as the research subjects.The therapeutic efficacy of 131I treatment in differentiated thyroid cancer patients was evaluated.91 patients were included in the 131I effective treatment group,and 40 patients with differentiated thyroid cancer were included in the 131I ineffective treatment group.The expression of TFCP2L1,DUSP26,and MLKL proteins in differentiated thyroid cancer was detected by immunohistochemistry.Multivariate Logistic regression analysis was used to analyze the influencing factors of ineffective 131I treatment in differentiated thyroid cancer patients.The predictive value of TFCP2L1,DUSP26,and MLKL for ineffective 131I treatment in differentiated thyroid cancer patients was evaluated by receiver operating characteristic(ROC)curve.Results In patients with differentiated thyroid carcinoma,the mRNA expression levels of TFCP2L1 and MLKL in tumor tissues were higher than those in adjacent non-tumor tissues,whereas DUSP26 mRNA expression was lower in tumor tissues(all P<0.05).The positive immunohistochemical expression rates of TFCP2L1 and MLKL were significantly higher in tumor tissues than in non-tumor tissues,while the positive rate of DUSP26 was significantly lower(all P<0.05).Expression levels of TFCP2L1,DUSP26,and MLKL were associated with the presence of multifocal disease and lymph node metastasis in differentiated thyroid carcinoma patients(all P<0.05).Compared with the 131I effective-treatment group,patients in the 131I ineffective-treatment group had higher proportions of multifocal disease,lymph node metastasis,131I dose>100 mCi,and higher levels of TFCP2L1 and MLKL,which were risk factors for 131I treatment ineffectiveness.DUSP26 levels were lower than in the effective group and acted as a protective factor(all P<0.05).The combined model of TFCP2L1,DUSP26,and MLKL for predicting 131I treatment effects in differentiated thyroid carcinoma yielded an area under the curve(AUC)of 0.950(95%CI:0.915-0.985),which was significantly superior to the AUCs of any single marker-TFCP2L1(Z=2.447,P<0.05),DUSP26(Z=2.382,P<0.05),or MLKL(Z=3.101,P<0.05).Conclusion The positive expression rates of TFCP2L1,MLKL in cancer tissue of patients with differentiated thyroid cancer were high,and the positive expression rates of DUSP26 were low,which were related to the clinicopathological characteristics of multifocal lesions and lymph node metastasis.The combined expression of the three factors had a high value in predicting the treatment effects of 131I in patients with differentiated thyroid cancer.

Abstract:Objective To investigate the efficacy of TC regimen(paclitaxel+carboplatin)plus bevacizumab combined with olaparib in the treatment of advanced ovarian cancer and its effect on immune function.Methods A total of 97 patients with ovarian cancer admitted to Nantong Tumor Hospital from June 2020 to June 2023 were selected as the research objects and divided into control group(n=48)and observation group(n=49)by random number table method.The control group was treated with bevacizumab combined with TC regimen,and the observation group was treated with olaparib on the basis of the control group.The clinical efficacy,serum carbohydrate antigen 199(CA199),carbohydrate antigen 125(CA125)and human epididymis protein 4(HE4)levels before treatment and at the end of the 2 nd,4 th and 6 th cycles of treatment,immune function and apoptosis factor levels before and after treatment were compared between the two groups.The adverse reactions during the treatment were observed,and the survival of the two groups was recorded after 1 year of follow-up.Results After treatment,the objective remission rate in the observation group was 69.39%,significantly higher than that in the control group(43.75%),and the difference were statistically significance(P<0.05),whereas no statistically significant difference was found in the disease control rates between the groups(81.63%vs.70.83%,P<0.05).The levels of serum CA199,CA125 and HE4 in the observation group were significantly lower than those in the control group at the end of the 2 nd,4 th and 6 th cycles of treatment(P<0.05).After treatment,the decrease of CD3+,CD4+,CD8+,natural killer(NK)cells,the increase of myeloid-derived suppressor cells(MDSCs),the levels of anti-apoptotic protein Bcl-2 and anti-apoptotic auxiliary protein Bag-1 in the observation group were significantly lower than those in the control group(P<0.05),and the level of pro-apoptotic protein Bax was significantly higher than that in the control group(P<0.05).During the treatment,the incidence of gastrointestinal reaction,hypodynamia,leukopenia and neutropenia in the control group was significantly higher than that in the observation group(P<0.05).The results of Kaplan-Meier survival curve showed that progression-free survival and overall survival in the observation group were significantly higher than those in the control group,and the differences were statistically significant(P<0.05).Conclusion TC regimen plus bevacizumab combined with olaparib in the treatment of ovarian cancer can enhance patients'short-term efficacy,significantly reduce the level of tumor markers,and decrease treatment-related adverse reactions by improving immune status and regulating cytokines,and significantly prolong the patients'survival.

Abstract:Objective To investigate the expression of NKG2D ligands(NKG2DLs)on leukemia cells in patients with acute myeloid leukemia(AML),and to preliminarily explore the safety and feasibility of umbilical cord blood-derived NKG2DL-targeting chimeric antigen receptor natural killer cells(CAR-NK)in the treatment of relapsed/refractory AML(RR-AML)patients.Methods Flow cytometry was used to detect NKG2DLs expression on tumor cells in AML patients admitted to the Affiliated Hospital of Jiangsu University between January 2023 and March 2024.Three patients with RR-AML were enrolled and treated with umbilical cord blood-derived NKG2D ligand-targeting CAR-NK cells following lymphodepleting chemotherapy.Results A total of 28 AML patients were enrolled in this study.Flow cytometric analysis of leukemic cells from AML patients revealed the surface expression of NKG2D ligands on the blasts.The median expression rates were as follows:1.72%for MICA/MICB,1.24%for ULBP-1,1.26%for ULBP-3,9.68%for ULBP-4,and 0.60%for ULBP-2/5/6.No statistically significant differences were observed in the median expression rates of NKG2D ligands between RR-AML and newly diagnosed AML patients(P>0.05).All three enrolled R/R AML patients received allogeneic CAR-NK cells at the predetermined dose post-lymphodepletion.Following infusion,CAR-NK cell expansion was detectable in all three patients.The peak level was reached 1 to 5 days after the first infusion,and the cells persisted for a maximum of 26 days.No significant coagulopathy,renal impairment,or graft-versus-host disease was observed in any of the three patients following CAR-NK cell infusion.However,one patient developed cytokine release syndrome and immune effector cell-associated neurotoxicity syndrome.In the CAR-NK treated cohort,two patients failed to respond,whereas one achieved complete remission.The responder successfully bridged to related donor haploidentical hematopoietic stem cell transplantation(HSCT),with subsequent rapid hematopoietic reconstitution and establishment of full donor chimerism.Conclusion NKG2DLs are expressed on the surface of leukemic cells in both newly diagnosed and RR-AML patients.Allogeneic umbilical cord blood-derived NKG2DL-targeting CAR-NK cells demonstrated an acceptable safety profile without new safety concerns upon infusion in RR-AML patients,although their persistence in vivo was relatively short-lived.Notably,RR-AML patients who responded to CAR-NK therapy could be successfully bridged to allogeneic HSCT.

Abstract:Objective To explore the role of the phosphatidy linositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway in the regulation of human gastric cell proliferation,apoptosis and 5-FU sensitivity by plumbagin(Plu).Methods Human gastric cancer cell line AGS was used as the research object.The cells were treated with low-dose Plu(2.5 μmol/L),high-dose Plu(5 μmol/L),and high-dose Plu combined with PI3K/AKT/mTOR pathway activator 740Y-P in vitro,with DMSO-treated cells serving as the control group.MTT assay,EdU staining and cell clone formation assay were used to detect cell proliferation ability.Annexin V-PI staining method was used to detect apoptosis in AGS cells.Western blotting was used to detect the protein expression and phosphorylation levels of key components in the PI3K/AKT/mTOR pathway in AGS cells.The 5-FU-resistant cell line AGS-R was induced and screened by the concentration-increasing method.After intervention with Plu and 740Y-P respectively,the changes in the resistance of AGS-R to 5-FU were detected to reflect the drug resistance of the cells.Results Compared with the control group,low and high doses of Plu treatment significantly reduced the colony formation rate,EdU positivity rate,p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR levels of AGS cells,and significantly increased the apoptosis rate(P<0.05).High-dose Plu combined with 740Y-P treatment significantly increased the colony formation rate,EdU positivity rate,p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR levels of AGS cells,and significantly decreased the apoptosis rate(P<0.05).The resistance index of 5-FU resistant AGS cells was 4.95,and the reversal fold was 2.05(>1.5).Plu intervention significantly reduced the EdU positive rate of AGS-R cells and significantly increased the apoptosis rate(P<0.05).However,co-administration of a high-dose Plu with 740Y-P significantly reversed these changes(P<0.05).Conclusion Plu inhibits the proliferation of human gastric cancer cells by down-regulating the activity of the PI3K/AKT/mTOR pathway,promotes cell apoptosis,and enhances the sensitivity to 5-FU.

Abstract:Objective To investigate the expression,prognostic value,and relationship with immune infiltration of RCSD domain-containing protein 1(RCSD1)in non-small cell lung cancer(NSCLC).Methods The expression difference of RCSD1 between NSCLC and normal tissues was analyzed using the Tumor Immune Estimation Resource(TIMER)and the Gene Expression Profiling Interactive Analysis databases,followed by experimental validation with real-time fluorescence quantitative PCR on 93 paired tumor and adjacent normal samples.Furthermore,the correlation between RCSD1 expression and clinicopathological characteristics was assessed.The Kaplan-Meier method and Cox proportional hazards model were employed to evaluate its prognostic value.Finally,the relationship between RCSD1 expression,its copy number variation,and the level of tumor immune cell infiltration was investigated using the TIMER database.Results Database analysis revealed that RCSD1 expression was significantly lower in both lung adenocarcinoma and lung squamous cell carcinoma tissues compared to normal tissues(P<0.05),and its expression level decreased progressively with advancing TNM stage(P=0.041).Experimental results confirmed that RCSD1 expression was lower in tumor tissues than in adjacent normal tissues(0.413±0.238 vs.1.172±0.277,P<0.001).Low RCSD1 expression was significantly associated with advanced TNM stage(Stage III:31.2%vs.13.3%,P=0.039),higher T stage(T3+T4:29.2%vs.11.1%,P=0.031),lymph node metastasis(N1+N2:60.4%vs.26.7%,P=0.001),tumor size>3 cm(62.5%vs.28.9%,P=0.001),and vascular invasion(41.7%vs.17.8%,P=0.012).The median disease-free survival(DFS)in the high RCSD1 expression group was 38.2(95%CI:32.5-43.9)months,significantly longer than the 25.6(95%CI:20.3-30.9)months in the low expression group(P=0.001).Multivariate analysis confirmed that low RCSD1 expression was an independent risk factor for DFS(HR=2.349,95%CI:1.249-4.432,P=0.008).RCSD1 expression showed a significant negative correlation with tumor purity(adenocarcinoma:r=-0.514,P=1.17e-34;squamous cell carcinoma:r=-0.482,P=3.07e-29)but was positively correlated with the infiltration levels of six types of immune cells including B cells,CD8+T cells,CD4+T cells,macrophages,neutrophils,and dendritic cells(adenocarcinoma:r=0.575-0.673,P<0.001;squamous cell carcinoma:r=0.512-0.799,P<0.001).Furthermore,RCSD1 copy number variation was notably associated with immune cell infiltration patterns,with dendritic cell infiltration being most prominent in the genomic amplification state.Conclusion Low RCSD1 expression is a significant prognostic factor in NSCLC,which is closely associated with tumor progression and immune cell infiltration,suggesting its potential as a novel biomarker and therapeutic target.

Abstract:Objective To discuss the impacts of long non-coding RNA OIP5-AS1(LncRNA OIP5-AS1)on the proliferation,migration and invasion of papillary thyroid carcinoma(PTC)cells by modulating the microRNA-410-3p(miR-410-3p)/non-muscle myosin heavy chain 9(MYH9)axis.Methods Real-time fluorescence quantitative PCR reaction(RT-qPCR)and Western blotting were used to detect the expression of LncRNA OIP5-AS1,miR-410-3p,and MYH9 in PTC cell lines(IHH4,TPC-1,B-CPAP)and normal human thyroid follicular epithelial cells Nthy-ori3-1,in order to screen for the best intervention cells.TPC-1 cells were separated into Control group,sh-NC group,sh-OIP5-AS1 group,sh-OIP5-AS1+anti-miR-NC group,sh-OIP5-AS1+anti-miR-410-3p group,mimics NC group,miR-410-3p mimics group,miR-410-3p mimics+pcDNA group,and miR-410-3p mimics+MYH9 group.The method of RT-qPCR was used to detect the expression of LncRNA OIP5-AS1 and miR-410-3p of cells.Plate cloning experiments and flow cytometry were used to detect cell proliferation and apoptosis,respectively.Transwell chamber experiment was used to measure cell migration and invasion.Western blotting was used to measure the protein expression of MYH9,Ki-67,MMP-2 and MMP-9.The dual luciferase reporter genes were used to measure the targeting relationship between LncRNA OIP5-AS1,MYH9,and miR-410-3p.The nude mouse transplant tumor experiment was used to verify the effect of LncRNA OIP5-AS1 on the growth of PTC transplant tumors.Results Compared with the Nthy-ori3-1 cell,the LncRNA OIP5-AS1 and MYH9 protein expression increased in IHH4,TPC-1,and B-CPAP cells,while miR-410-3p expression decreased(P<0.05).TPC-1 cells were selected for the experiment.Silencing LncRNA OIP5-AS1 or overexpression of miR-410-3p was able to prominently upregulate miR-410-3p expression and downregulate MYH9 expression,reduce the number of clone formation,cell migration and invasion,and increase the expression of Ki-67,MMP-2,and MMP-9 proteins,and also increase the apoptosis rate of cells(P<0.05).Inhibition of miR-410-3p or overexpression of MYH9 was able to weaken the effect of silencing LncRNA OIP5-AS1 or overexpression of miR-410-3p on the malignant behaviors of TPC-1 cells(P<0.05).LncRNAs OIP5-AS1,MYH9 had a targeted regulatory relationship with miR-410-3p.Silencing LncRNA OIP5-AS1 could significantly reduce the volume,mass and expression levels of LncRNA OIP5-AS1 and MYH9 in the PTC transplanted tumors,and significantly increase the expression of miR-410-3p(P<0.05).Conclusion Silencing LncRNA OIP5-AS1 can inhibit the malignant progression of PTC by modulating miR-410-3p/MYH9 axis.

Abstract:In recent years,the application of patient self-reported outcomes(PROs)in clinical trials of cancer drugs has gradually become a research hotspot.PROs have become an important supplement to traditional medical indicators and play a key role in quality of life assessment,symptom monitoring,risk prediction,clinical decision support,and new drug approval.However,the current application of PROs still faces problems such as scale selection,data collection and management,data interpretation and standardization,which limits their comprehensive promotion in clinical practice.This paper aims to systematically sort out the application status,problems,and improvement direction of PROs in clinical trials of cancer drugs,and provide a theoretical basis and practical reference for researchers in the field of oncology drug clinical trials in China,so as to further improve the safe and quality of life of patients.