Objective To investigate the impact of Toxoplasma gondii type I, II and III rhoptry protein 16 (ROP16) on programmed cell death ligand 1 (PD-L1) expression in lung adenocarcinoma cells, and to examine the effects of T. gondii type I ROP16 protein on the relative PD-L1 expression, the relative PD-L1 distribution on the cell membrane surface, and the binding of programmed cell death 1 (PD-1) to PD-L1 in lung adenocarcinoma cells. Methods Lentiviral vectors overexpressing T. gondii type I, II and III ROP16 proteins were generated, and transfected into the human lung adenocarcinoma A549 cell line. A549 cells were used as a blank control group, and A549 cells transfected with an empty lentiviral expression vector were used as a negative control group, while A549 cells transfected with lentiviral vectors overexpressing T. gondii type I, II and III ROP16 proteins served as experimental groups. Stably transfected cells were selected with puromycin and verified using Western blotting, quantitative real-time PCR (RT-qPCR), and immunofluorescence assays. The PD-L1 expression was quantified at translational and transcriptional levels using Western blotting and RT-qPCR assays in A549 cells in the five groups, and the relative PD-L1 distribution was detected on the A549 cell membrane surface using flow cytometry. In addition, the effect of T. gondii type I ROP16 protein on the PD-1/PD-L1 binding was measured in A549 cells using enzyme-linked immunosorbent assay (ELISA). Results The relative ROP16 protein expression was 0, 0, 1.546 ± 0.091, 1.822 ± 0.047 and 2.334 ± 0.089 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 1 339.00,P < 0.001), and the relative ROP16 mRNA expression was 2.153 ± 0.949, 2.436 ± 1.614, 14.343 ± 0.020, 12.577 ± 0.285 and 15.090 ± 0.420 in the blank control group, negative control group and the T. gondii type I, II and III ROP16 protein overexpression groups, respectively (F = 483.50,P < 0.001). The ROP16 expression was higher in the T. gondii type I, II and III ROP16 protein overexpression groups than in the blank control group at both translational and transcriptional levels (allP values < 0.001). Immunofluorescence assay revealed that T. gondii type I, II and III ROP16 proteins were predominantly localized in A549 cell nuclei. Western blotting showed that the relative PD-L1 protein expression was 0.685 ± 0.109, 0.589 ± 0.114, 1.007 ± 0.117, 0.572 ± 0.151, and 0.426 ± 0.116 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 9.46,P < 0.05), and RT-qPCR assay quantified that the relative PD-L1 mRNA expression was 1.012 ± 0.190, 1.281 ± 0.465, 1.950 ± 0.175, 0.889 ± 0.251, and 0.230 ± 0.192 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 14.18,P < 0.05). The PD-L1 expression was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group at both translational and transcriptional levels (both P values < 0.05). Flow cytometry detected that the relative distributions of PD-L1 protein were (10.83 ± 0.60)%, (11.23 ± 0.20)%, and (14.61 ± 0.50)% on the A549 cell membrane surface (F = 28.31, P < 0.05), and the relative distribution of PD-L1 protein was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group and negative control group (both P values < 0.001). ELISA measured significant differences in the absorbance (A) value among the T. gondii type IROP16 protein overexpression group, the blank control group and the negative control group if the concentrations of the recombinant PD-1 protein were 0.04 (F = 10.45, P < 0.05), 0.08 μg/mL (F = 11.68, P < 0.05) and 0.12 μg/mL (F = 52.68, P < 0.05), and the A value was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group and the negative control group (both P values < 0.05), indicating that T. gondii type IROP16 protein promoted the PD-L1/PD-1 binding in A549 cells in a concentration-dose manner. Conclusions T. gondii type IROP16 protein overexpression may up-regulate PD-L1 expression in A549 cells at both transcriptional and translational levels and the relative PD-L1 distribution on the A549 cell membrane surface, and affect the PD-1/PD-L1 binding in a concentration-dependent manner.

Objective:To analyze the endoscopic and clinicopathological characteristics of Helicobacter pylori ( HP)-negative early gastric cancer (EGC). Methods:A retrospective study was conducted on data of patients who were diagnosed as having EGC at the Affiliated Hospital of Qingdao University between June 2013 and March 2024. Cases meeting the diagnostic criteria for HP-negative status were included. Endoscopic findings and histopathological features of HP-negative EGC were systematically analyzed. Results:Among 698 EGC patients, 29 (4.2%) were identified as HP-negative. The age at diagnosis was 59.9±10.0 years, with marked female predominance (69.0%, 20/29 VS 31.0% males, 9/29). A body mass index ≥24 kg/m 2 was observed in 69.0% (20/29). Regarding therapeutic approaches, endoscopic submucosal dissection was performed in 89.7% (26/29). Among the 29 patients with a total of 30 lesions, the majority were localized to the middle third of the stomach (40.0%, 12/30), or the upper third (36.7%, 11/30). Differentiated-type accounted for 73.3% (22/30) among the histological types, including 13 oxyntic gland adenoma (OGA) / gastric adenocarcinoma of fundic-gland type (GA-FG) (upper third: 7; middle third: 6), and 2 gastric adenocarcinomas of fundic-gland mucosa type (GA-FGM) (both upper third). White-light endoscopy revealed polypoid or small submucosal tumor-like protrusions with whitish or erythematous discoloration and characteristic branching dilated vessels on the surface. Among the OGA/GA-FG lesions, 8 exhibited indistinct demarcation lines (DL). Additionally, both GA-FGM lesions demonstrated poorly defined DL. Three gastric adenocarcinomas of foveolar-type (GA-FV) were identified (upper/middle/lower third: 1 each), with 2 presenting as erythematous elevated masses. Five signet ring cell carcinomas (upper/middle/lower third: 1/3/1) exhibited flat or shallow depressed morphology with whitish or erythematous discoloration under white-light endoscopy; 3 exhibited well-demarcated borders. Four pyloric gland adenocarcinomas and three poorly differentiated tubular adenocarcinomas showed no significant differences in endoscopic features on white-light and narrow-band imaging compared to HP-positive EGC. No intestinal-type adenocarcinomas were identified. Conclusion:HP-negative EGC exhibit distinct endoscopic and clinicopathological characteristics, including female predominance and frequent differentiated histology, and upper/middle-third localization of elevated lesions, primarily OGA/GA-FG.

Objective:This study analyzes the clinicopathological features and prognostic factors of early-stage gastric mixed adenoneuroendocrine carcinoma (G-MANEC), which is an exceedingly rare malignancy, in an effort to provide evidence-based insights for clinical decision-making.Methods:A retrospective observational study was conducted using the clinical data of 36 patients with early-stage G-MANEC who underwent surgical resection at Zhongshan Hospital, Fudan University, from July 2014 to May 2022. The observed indicators included clinicopathological data and follow-up information on recurrence, metastasis, and overall survival (OS).Results:Among the 36 patients there were 21 males and 15 females, aged 32-84 (65±11) years. The most common initial symptoms were abdominal pain and distension (19/36, 52.8%), followed by incidental findings during physical examinations (7/36, 19.4%). Tumors were located in the proximal stomach in 13 cases (36.1%), the middle stomach in 4 cases (11.1%), and the distal stomach in 19 cases (52.8%). Average tumor diameter was (2.48±1.18) cm. Gross morphology included elevated type in 12 cases (33.3%), flat type in 20 cases (55.6%), and depressed type in 4 cases (11.1%). Ulceration was present in 12 cases (33.3%). There were 11 cases (30.6%) at T1a stage and 25 cases (69.4%) at T1b stage. Lymph node metastasis was positive in 10 cases (27.8%), and the differentiation grades of the adenocarcinoma component were Grade I, II, and III in 3 (8.3%), 10 (27.8%), and 23 (63.9%) cases, respectively. Furthermore, the proportion of neuroendocrine carcinoma component was ≥50% in 18 cases (50.0%) and <50% in 18 cases (50.0%). Lymphovascular or perineural invasion was present in 18 cases (50.0%). Lauren classification included mixed type in 10 cases (27.8%), intestinal type in 19 cases (52.8%), and diffuse type in 7 cases (19.4%), and chromogranin A (CgA) positivity was found in 20 cases (55.6%). Additionally, the Ki-67 index positivity was found in 26 cases (72.2%). Total gastrectomy was performed in 12 cases (33.3%) and partial gastrectomy in 24 cases (66.7%), with a median follow-up duration of 77.5 months. The 3-year and 5-year OS rates were 88.89% and 79.67%, respectively. Univariate analysis revealed that age, gross morphology, ulceration, proportion of neuroendocrine carcinoma component, lymphovascular or perineural invasion, and chromogranin A (CgA) positivity showed statistical significance in their association with OS ( P<0.10). Multivariate Cox regression analysis further identified ulceration (HR=7.74, 95%CI: 1.24-48.30, P=0.028) and CgA positivity (HR=21.76, 95%CI: 1.86-53.97, P=0.014) as independent risk factors of OS. Conclusions:Patients with early-stage G-MANEC are typically asymptomatic, and those with ulceration or positive CgA immunohistochemical staining tend to have a poor prognosis.

Objective:This study analyzes the clinicopathological features and prognostic factors of early-stage gastric mixed adenoneuroendocrine carcinoma (G-MANEC), which is an exceedingly rare malignancy, in an effort to provide evidence-based insights for clinical decision-making.Methods:A retrospective observational study was conducted using the clinical data of 36 patients with early-stage G-MANEC who underwent surgical resection at Zhongshan Hospital, Fudan University, from July 2014 to May 2022. The observed indicators included clinicopathological data and follow-up information on recurrence, metastasis, and overall survival (OS).Results:Among the 36 patients there were 21 males and 15 females, aged 32-84 (65±11) years. The most common initial symptoms were abdominal pain and distension (19/36, 52.8%), followed by incidental findings during physical examinations (7/36, 19.4%). Tumors were located in the proximal stomach in 13 cases (36.1%), the middle stomach in 4 cases (11.1%), and the distal stomach in 19 cases (52.8%). Average tumor diameter was (2.48±1.18) cm. Gross morphology included elevated type in 12 cases (33.3%), flat type in 20 cases (55.6%), and depressed type in 4 cases (11.1%). Ulceration was present in 12 cases (33.3%). There were 11 cases (30.6%) at T1a stage and 25 cases (69.4%) at T1b stage. Lymph node metastasis was positive in 10 cases (27.8%), and the differentiation grades of the adenocarcinoma component were Grade I, II, and III in 3 (8.3%), 10 (27.8%), and 23 (63.9%) cases, respectively. Furthermore, the proportion of neuroendocrine carcinoma component was ≥50% in 18 cases (50.0%) and <50% in 18 cases (50.0%). Lymphovascular or perineural invasion was present in 18 cases (50.0%). Lauren classification included mixed type in 10 cases (27.8%), intestinal type in 19 cases (52.8%), and diffuse type in 7 cases (19.4%), and chromogranin A (CgA) positivity was found in 20 cases (55.6%). Additionally, the Ki-67 index positivity was found in 26 cases (72.2%). Total gastrectomy was performed in 12 cases (33.3%) and partial gastrectomy in 24 cases (66.7%), with a median follow-up duration of 77.5 months. The 3-year and 5-year OS rates were 88.89% and 79.67%, respectively. Univariate analysis revealed that age, gross morphology, ulceration, proportion of neuroendocrine carcinoma component, lymphovascular or perineural invasion, and chromogranin A (CgA) positivity showed statistical significance in their association with OS ( P<0.10). Multivariate Cox regression analysis further identified ulceration (HR=7.74, 95%CI: 1.24-48.30, P=0.028) and CgA positivity (HR=21.76, 95%CI: 1.86-53.97, P=0.014) as independent risk factors of OS. Conclusions:Patients with early-stage G-MANEC are typically asymptomatic, and those with ulceration or positive CgA immunohistochemical staining tend to have a poor prognosis.

Objective:To analyze the endoscopic and clinicopathological characteristics of Helicobacter pylori ( HP)-negative early gastric cancer (EGC). Methods:A retrospective study was conducted on data of patients who were diagnosed as having EGC at the Affiliated Hospital of Qingdao University between June 2013 and March 2024. Cases meeting the diagnostic criteria for HP-negative status were included. Endoscopic findings and histopathological features of HP-negative EGC were systematically analyzed. Results:Among 698 EGC patients, 29 (4.2%) were identified as HP-negative. The age at diagnosis was 59.9±10.0 years, with marked female predominance (69.0%, 20/29 VS 31.0% males, 9/29). A body mass index ≥24 kg/m 2 was observed in 69.0% (20/29). Regarding therapeutic approaches, endoscopic submucosal dissection was performed in 89.7% (26/29). Among the 29 patients with a total of 30 lesions, the majority were localized to the middle third of the stomach (40.0%, 12/30), or the upper third (36.7%, 11/30). Differentiated-type accounted for 73.3% (22/30) among the histological types, including 13 oxyntic gland adenoma (OGA) / gastric adenocarcinoma of fundic-gland type (GA-FG) (upper third: 7; middle third: 6), and 2 gastric adenocarcinomas of fundic-gland mucosa type (GA-FGM) (both upper third). White-light endoscopy revealed polypoid or small submucosal tumor-like protrusions with whitish or erythematous discoloration and characteristic branching dilated vessels on the surface. Among the OGA/GA-FG lesions, 8 exhibited indistinct demarcation lines (DL). Additionally, both GA-FGM lesions demonstrated poorly defined DL. Three gastric adenocarcinomas of foveolar-type (GA-FV) were identified (upper/middle/lower third: 1 each), with 2 presenting as erythematous elevated masses. Five signet ring cell carcinomas (upper/middle/lower third: 1/3/1) exhibited flat or shallow depressed morphology with whitish or erythematous discoloration under white-light endoscopy; 3 exhibited well-demarcated borders. Four pyloric gland adenocarcinomas and three poorly differentiated tubular adenocarcinomas showed no significant differences in endoscopic features on white-light and narrow-band imaging compared to HP-positive EGC. No intestinal-type adenocarcinomas were identified. Conclusion:HP-negative EGC exhibit distinct endoscopic and clinicopathological characteristics, including female predominance and frequent differentiated histology, and upper/middle-third localization of elevated lesions, primarily OGA/GA-FG.

Objective To observe the value of ultrasonic parameters for predicting whether patients with cesarean scar pregnancy(CSP)would benefit from ultrasound-guided suction curettage alone.Methods Totally 140 CSP patients diagnosed by transvaginal ultrasound and initially treated with ultrasound-guided suction curettage alone were prospectively recruited and categorized into benefited group(n=103)and non-benefited group(n=37)according to bleeding during suction curettage and prognoses.The ultrasonic manifestations of CSP were observed,and the thickness of chorionic villi at the scar,as well as of residual myometrium of the anterior wall in the lower segment of the uterus,also the maximum diameter of the gestational sac were measured and compared between groups,and the parameters with quantitative data being statistically different between groups were converted into categorical predictor through analyzing of the receiver operating characteristic(ROC)curves and the optimal cut-off values.The independent predictors were selected among ultrasonic features and categorical predictor variables being statistically different between groups using multivariate logistic regression,and a combined predicting model was then constructed,and the predicting efficacy of the combined model and each categorical predictor alone was assessed according to the area under curve(AUC)and then were compared.Results Compared with non-benefited group,the gestational weeks in benefited group were smaller(P＜0.05),and the percentages of rich blood supply and the presence of embryos and fetal heartbeats were lower,with thinner chorionic villi at the scar,thicker residual myometrium and smaller maximal diameter of the gestational sac in benefited group(all P＜0.05).ROC curves analyses yielded the best cut-off value for dichotomy of chorionic villi thickness at the scar was 4.7 mm,of residual myometrium thickness was 1.8 mm and of the maximum diameter of the gestational sac was 29 mm,respectively,and then categorical predictor variable were obtained.Multivariate logistic regression showed that the transformed categorical predictors,i.e.the thickness degree of the chorionic villi at the scar,the thickness degree of the residual muscle layer and the size degree of the gestational sac,were all independent predictors of whether CSP patients would benefit from ultrasound-guided suction curettage alone(all P＜0.05).The AUC of the combined predicting model was 0.918,higher than that of each transformed categorical predictor alone(all P＜0.05).Conclusion The thickness of the chorionic villi at the scar ≤4.7 mm,the thickness of the residual muscle layer＞1.8 mm and the maximum diameter of the gestational sac≤29 mm were all independent predictors of CSP patients would benefit from ultrasound-guided suction curettage alone,and the predicting efficacy of the combined model was higher than that of each alone.

Although immunotherapy has made a breakthrough in the treatment of cancer,the emergence of drug resistance has limited be-nefits in most patients,and reversing immunotherapy resistance remains a hotly debated and unresolved issue.The tumor microenviron-ment(TME)comprises a large number of T lymphocytes.During the biological processes of proliferation,activation,and the effect of tumor-infiltrating lymphocytes(TILs),TILs face severe local metabolic stress,resulting in metabolic reprogramming to meet the markedly increased energy demand and biosynthesis requirements.This process of adaptive changes in the metabolic pattern is closely related to the pheno-type and function of TILs,affecting immunotherapy efficacy and mediating immunotherapy resistance.In recent years,with the study of TIL metabolism and the search for specific metabolic regulatory points,targeting the TIL metabolic pathway could restore the tumor-killing func-tion and reverse immunotherapy resistance.This review explores the relationship between the metabolic reprogramming of TILs and im-mune resistance,and provides a new strategy for reversing immune resistance.

Objective:To investigate the risk factors for lymph node metastasis in patients with early gastric cancer and establish a model for prediction of risk.Methods:The cohort of this retrospective observational study comprised 1096 patients who had undergone radical gastric cancer surgery combined with standard D1 lymphadenectomy and been diagnosed with early gastric cancer by postoperative pathology in Zhongshan Hospital affiliated with Fudan University from January 2016 to July 2022. The patients were allocated to groups with and without lymph node metastases. Clinicopathological characteristics were compared between the two groups and multi-factor logistic regression analysis used to identify independent risk factors for lymph node metastasis in patients with early gastric cancer. Indications for endoscopic resection in the Japanese Gastric Cancer Association (JGCA) guideline were also incorporated into construction of the model. The patient cohort was divided into training and validation sets in a 6:4 ratio. The identified independent risk factors were used to construct a predictive nomogram. Receiver operating characteristic curves were plotted separately and the difference between them in predictive efficacy was compared using the area under the curve (AUC).Results:A total of 1,096 patients with early gastric cancer were included, with 750 males and 346 females. Their average age was (61.4±10.9) years old, and the mean tumor diameter was (23.8±11.4) mm. Among them, 188 patients (17.2%) had positive lymph node metastasis, with 109 cases in N1 stage, 42 cases in N2 stage, and 37 cases in N3 stage. Additionally, 462 patients were in T1a stage, while 634 patients were in T1b stage. Univariate analysis showed that tumor diameter, location, Lauren classification, gross morphology, histological type, intravascular invasion, ulceration, differentiation type and tumor T stage were associated with lymph node metastasis after radical gastrectomy for early gastric cancer (all P<0.05). Multifactorial analysis showed that the presence of intravascular invasion (OR=14.822, 95%CI: 9.323–23.572, P<0.001), undifferentiated type (OR=3.095, 95%CI: 1.649–5.811, P<0.001), tumor T1b (OR=1.798, 95%CI: 1.053–3.079, P=0.032), and tumor diameter ≥2 cm (OR=1.229, 95%CI: 1.031–1.469, P=0.022) were independent risk factors for lymph node metastasis. The baseline data of the training set and validation set were consistent in terms of balance (all P>0.05). We used the above variables to establish a predictive nomogram for lymph node metastasis in patients with early gastric cancer. The AUC values obtained from the validation of the model in the training and validation sets were 0.880 (95%CI: 0.849–0.911) and 0.881 (95%CI: 0.841–0.921), respectively, and were significantly better than the predictive efficacy based on the JGCA guideline (AUC=0.777, 95%CI: 0.746–0.809, P<0.001). Conclusions:Patients with early gastric cancer and intravascular invasion, undifferentiated tumors, tumor T1b, and diameter ≥2 cm are at higher risk of postoperative lymph node metastasis than other patients. The predictive model developed in this study more accurately predicts lymph node metastasis in patients with early gastric cancer than previously proposed methods.

Objective:To investigate the risk factors for lymph node metastasis in patients with early gastric cancer and establish a model for prediction of risk.Methods:The cohort of this retrospective observational study comprised 1096 patients who had undergone radical gastric cancer surgery combined with standard D1 lymphadenectomy and been diagnosed with early gastric cancer by postoperative pathology in Zhongshan Hospital affiliated with Fudan University from January 2016 to July 2022. The patients were allocated to groups with and without lymph node metastases. Clinicopathological characteristics were compared between the two groups and multi-factor logistic regression analysis used to identify independent risk factors for lymph node metastasis in patients with early gastric cancer. Indications for endoscopic resection in the Japanese Gastric Cancer Association (JGCA) guideline were also incorporated into construction of the model. The patient cohort was divided into training and validation sets in a 6:4 ratio. The identified independent risk factors were used to construct a predictive nomogram. Receiver operating characteristic curves were plotted separately and the difference between them in predictive efficacy was compared using the area under the curve (AUC).Results:A total of 1,096 patients with early gastric cancer were included, with 750 males and 346 females. Their average age was (61.4±10.9) years old, and the mean tumor diameter was (23.8±11.4) mm. Among them, 188 patients (17.2%) had positive lymph node metastasis, with 109 cases in N1 stage, 42 cases in N2 stage, and 37 cases in N3 stage. Additionally, 462 patients were in T1a stage, while 634 patients were in T1b stage. Univariate analysis showed that tumor diameter, location, Lauren classification, gross morphology, histological type, intravascular invasion, ulceration, differentiation type and tumor T stage were associated with lymph node metastasis after radical gastrectomy for early gastric cancer (all P<0.05). Multifactorial analysis showed that the presence of intravascular invasion (OR=14.822, 95%CI: 9.323–23.572, P<0.001), undifferentiated type (OR=3.095, 95%CI: 1.649–5.811, P<0.001), tumor T1b (OR=1.798, 95%CI: 1.053–3.079, P=0.032), and tumor diameter ≥2 cm (OR=1.229, 95%CI: 1.031–1.469, P=0.022) were independent risk factors for lymph node metastasis. The baseline data of the training set and validation set were consistent in terms of balance (all P>0.05). We used the above variables to establish a predictive nomogram for lymph node metastasis in patients with early gastric cancer. The AUC values obtained from the validation of the model in the training and validation sets were 0.880 (95%CI: 0.849–0.911) and 0.881 (95%CI: 0.841–0.921), respectively, and were significantly better than the predictive efficacy based on the JGCA guideline (AUC=0.777, 95%CI: 0.746–0.809, P<0.001). Conclusions:Patients with early gastric cancer and intravascular invasion, undifferentiated tumors, tumor T1b, and diameter ≥2 cm are at higher risk of postoperative lymph node metastasis than other patients. The predictive model developed in this study more accurately predicts lymph node metastasis in patients with early gastric cancer than previously proposed methods.

Henoch-Schonlein purpura (HSP) is one of the most common clinical manifestations of systemic small vasculitis in children with non-thrombocytopenic, and mainly manifested as skin purpura, arthritis, gastrointestinal symptoms and Henoch-Schonlein purpura nephritis (HSPN). The severity of renal involvement is the main factor determining the long-term prognosis of children with HSP.Studies have revealed that the determination of pentraxin 3 (PTX3) in serum can be used for early diagnosis of HSPN and prediction of renal injury.In this paper, the origin, gene and protein structure, function, potential relationship and mechanism of action between PTX3 and HSP were discussed, so as to provide new ideas for the early diagnosis and treatment of HSPN.