Tumor neoantigens are antigens encoded by tumor-specific mutated genes, characterized by high specificity, significant exogenous origin, mutation randomness, clonal distribution and correlation with gene mutation. Because these antigens are not negatively screened by the thymus and recognized by T cells as "heterogeneous". They are less easily affected by the immune tolerance mechanism and exhibit strong immunogenicity, making them excellent targets for immunotherapy. Tumor neoantigens can be used to develop therapeutic vaccines, induce and cultivate T cells with stronger targeting capabilities, and are promising for predicting tumor survival prognosis and responses to immune checkpoint blockade therapies. This review summarizes the recent advances in clinical application of tumor neoantigen-based immunotherapy, and prospects for future research directions.

Objective To investigate the prognosis of patients with hepatitis B virus (HBV)‑related intrahepatic cholangiocarcinoma (ICC) whose HBV DNA was negative before surgical. Methods A retrospective analysis was conducted on the clinical data of 97 ICC patients who underwent surgery resection at the Fifth Medical Center of Chinese PLA General Hospital between October 2010 and January 2017. All patients were divided into HBV-related ICC (HBV-ICC) group (n=62) and non-HBV-related ICC (Con-ICC) group (n=35). HBV-ICC group included 34 patients with HBV core antigen positive (HBcAb⁺) and HBV surface antigen positive (HBsAg⁺), and 28 patients with HBcAb positive and HBsAg negative. Kaplan-Meier analysis was used to plot survival curves and compare the overall survival (OS) and postoperative recurrence-free survival (RFS) among patients in Con-ICC, ICC patients with HBsAg⁺/HBcAb⁺, and ICC patients with HBsAg^-/HBcAb⁺. Univariate and multivariate Cox proportional hazard models were used to analyze independent influencing factor for OS, RFS and early postoperative recurrence among gender, age, pathogenic factor, liver cirrhosis, Child-Pugh grade, carbohydrate antigen 19-9 (CA199), alpha-fetoprotein (AFP), glutamine transferase (GGT), alkaline phosphatase (ALP), total bilirubin (TBil), direct bilirubin (DBil), American Joint Committee on Cancer (AJCC) stage, tumor size, tumor number, tumor differentiation, microvascular invasion, lymph node metastasis, hepatectomy procedure, cholecystectomy, and follow-up treatment. Results Of the 97 patients, the median age was 56 years, and 79 (81.4%) of them were male. The median follow-up time was 92.2 months. Eighty-eight (90.7%) patients presented with tumor recurrence and 73 (75.3%) died. In multivariate analyses, HBV-ICC and CA199>37 kU/L were independent predictors of OS (HR=0.45, 95%CI 0.26-0.77, P=0.003; HR=2.10, 95%CI 1.24-3.57, P=0.006), RFS (HR=0.43, 95%CI 0.27-0.68, P<0.001; HR=1.78, 95%CI 1.12-2.81, P=0.014), and postoperative early recurrence (HR=0.42, 95%CI 0.26-0.70, P=0.001; HR=2.02, 95%CI 1.20-3.39, P=0.008). AJCC stage Ⅲ was an independent risk factor for postoperative RFS (HR=1.81, 95%CI 1.04-3.14, P=0.037). Multiple tumor lesions was an independent risk factor for postoperative RFS and early recurrence (HR=1.73, 95%CI 1.07-2.77, P=0.024; HR=1.90, 95%CI 1.12-3.24, P=0.017). There was no statistically significant difference in OS, RFS, and early recurrence between HBV-ICC patients with HBsAg^-/HBcAb⁺ and Con-ICC patients (P<0.05), whereas HBsAg⁺/HBcAb⁺ was a significant factor affecting postoperative OS (HR=0.32, 95%CI 0.16-0.62, P=0.001), RFS (HR=0.32, 95%CI 0.18-0.55, P<0.001), and early recurrence (HR=0.29, 95%CI 0.15-0.54, P<0.001) in ICC patients. Conclusions The prognosis of HBV-ICC patients with preoperative HBV-DNA^- is better than that of Con-ICC patients. The prognosis of HBV-ICC patients with HBcAb⁺/HBsAg^- is worse than that of HBV-ICC patients with HBcAb⁺/HBsAg⁺, but similar to Con-ICC patients. Therefore, the postoperative stratified management of HBV-ICC patients should be emphasized.

Objective To investigate the relationship between uric acid metabolism and brain injury following cardiopulmonary bypass (CPB) in rats. Methods Healthy male SD rats were randomly assigned to either a Sham group or a CPB group, each comprising 12 rats. The Sham group only underwent vascular puncture and did not perform CPB conversion, while the CPB group was subjected to a CPB procedure with a perfusion duration of 110 min, and the brain tissue was collected post-procedure. Microdialysate was collected 1 h before and after CPB initiation. Apoptosis in the paraventricular nucleus (PVN) was assessed using TUNEL staining, and the expression of Bax mRNA in cerebral cortex and hypothalamus was determined via real-time quantitative PCR. Apoptosis-related protein expression was analyzed by Western blotting. Differentially expressed genes (DEGs) were identified through RNA-sequencing between brain tissues of two groups, and Gene Ontology (GO) analysis was performed to identify enriched pathways among the DEGs. Protein-protein interaction (PPI) networks were constructed using String and Cytoscape softwares to identify key genes. Liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed to analyze differential metabolites in the PVN before and after CPB, with Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis constructed subsequently. Uric acid levels in the hypothalamus was measured using a uric acid assay kit, and the expression of key enzymes of uric acid metabolism [xanthine reductase (XDH), adenosine deaminase (ADA)] and uric acid transporter [organic anion transporter family protein 1 (OAT1), organic anion transporter family protein 3 (OAT3), ATP-binding cassette transporter subfamily G member 2 (ABCG2), glucose transporter 9 (GLUT9)] genes in the hypothalamus was evaluated by real-time quantitative PCR. Results Real-time quantitative PCR revealed a significant upregulation of Bax mRNA in the cerebral cortex and hypothalamus of CPB group compared to Sham group (P<0.05). TUNEL staining indicated a significantly higher apoptosis rate of cells in PVN region in CPB group than that in Sham group (19.0%±5.0% vs. 7.6%±0.8%, P=0.01). Western blotting showed a significantly increased Bcl-2/Bax ratio in the hypothalamus of CPB group compared to Sham group (P<0.05). A total of 2829 DEGs were identified between Sham group and CPB group, with 1374 upregulated genes and 1455 downregulated genes. Uric acid metabolism-related pathways were predominantly enriched in purine nucleoside metabolism and biosynthesis, purine nucleoside monophosphate metabolism, purine nucleoside triphosphate metabolism, purine ribonucleotide metabolism and biosynthesis, purine ribonucleoside monophosphate metabolism and biosynthesis, purine ribonucleoside triphosphate metabolism and biosynthesis, and reaction to purine compounds. Eighteen differential metabolites were identified in the microdialysate, with 13 upregulated and 5 downregulated metabolites. KEGG enrichment analysis identified 7 significantly enriched metabolic pathways, among which the nicotinate and nicotinamide metabolism pathways were closely related to uric acid metabolism. Both RNA-sequencing and LC-MS/MS analysis suggested alterations in uric acid metabolism in CPB groups. Post-CPB, uric acid concentration in the hypothalamic tissue significantly increased (P<0.01), and the expression of XDH and ADA mRNA in the hypothalamus were significantly increased (P<0.05), while the expression of ABCG2, OAT1, OAT3 and GLUT9 mRNA significantly decreased (P<0.001). Conclusion Uric acid metabolism in brain is altered during CPB, which may be an important mechanism for brain injury following CPB.

Objective To investigate the molecular mechanism of verbascoside against acute lung injury (ALI) by network pharmacology and molecular docking methods, and to validate the findings experimentally. Methods The 2D structure of verbascoside was obtained from the Pubchem database. Active ingredient targets of verbascoside were acquired from Pharmmapper database and Swiss Target Prediction database. Active component targets of ALI were acquired from datebase such as Gene Cards, OMIM, and DisGeNET. Common targets between verbascoside and ALI were determined by overlapping these sets. PPI network for potential targets was constructed using String database and Cytoscape software. The intersection targets were imported into the DAVID database for enrichment analysis of GO biological processes, KEGG signaling pathway and the pathway target genes. Molecular docking between verbascoside and core targets was performed using Autodock vina software. The mRNA expression level of core genes was validated using real-time quantitative PCR (RT-qPCR), and the expression of related proteins was detected using Western blotting. Results A total of 150 target genes of verbascoside against ALI were screened, and the key targets of verbascoside against ALI mainly involve pathways such as Rap1 signaling pathway, PI3K-Akt signaling pathway and MAPK signaling pathway. Verbascoside docked well with the core target molecules. RT-qPCR results showed that, compared with the control group, the mRNA expression levels of HSP90AA1, ALB, TP53, TNF, INS, and HRAS were significantly decreased in cells after the effect of verbascoside (P<0.05); Western blotting indicated that, compared with the model group, verbascoside treatment significantly reduced the expression of p-Akt, p-p38, and p-ERK proteins (P<0.05). Conclusion Verbascoside could inhibit MAPK, Rap1 and PI3K/Akt signaling pathways to exert its anti-ALI effects.

Objective To compare the fidelity of chronic obstructive pulmonary disease (COPD) models established using two methods: exposure to cigarette smoke (CS) and exposure to motor vehicle exhaust (MVE) in rats. Methods Twenty-four male SD rats were randomly divided into control, CS-exposed (CS), and MVE-exposed (MVE) groups, with 8 rats per group. Rats in CS and MVE groups were exposed to CS or MVE, respectively, to induce COPD models. After COPD model established, lung function of each group was assessed. Bronchoalveolar lavage fluid (BALF) was collected to measure inflammatory cell counts, levels of inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor (TNF)‑α, and expression levels of mucin 5AC (MUC5AC). Lung tissue sections were stained with hematoxylin and eosin (HE) to observe pulmonary tissue and airway pathological changes. Periodic acid-Schiff (PAS) staining was used to detect goblet cell hyperplasia in airways. Results Compared with control group, rats in CS and MVE groups showed significantly increased inspiratory resistance (RI), total lung capacity (TLC), and lung static compliance (Cchord) (P<0.05), while expiratory flow parameters FEV₅₀/FVC were significantly decreased (P<0.05). Compared with MVE group, rats in CS group had significantly higher R_I, TLC, and Cchord (P<0.05), and lower FEV₅₀/FVC (P<0.05). HE staining of lung tissues showed that mean linear intercept (MLI) was significantly higher in both CS and MVE groups compared with control group (P<0.05), with CS group having higher MLI than MVE group (P<0.05). BALF analysis revealed that white blood cells, neutrophils, macrophages, lymphocytes, IL-6, and TNF-α levels were significantly higher in both CS and MVE groups compared with control group (P<0.05), and inflammatory cell counts, IL-6, and TNF-α levels were higher in CS group compared with MVE group(P<0.05). PAS staining of lung tissues indicated that goblet cells in large airways were significantly increased in both CS and MVE groups compared with control group (P<0.05), with CS group showing higher goblet cell counts than MVE group (P<0.05). Expression levels of MUC5AC in BALF were significantly higher in both CS and MVE groups compared with control group (P<0.05), with CS group having significantly higher MUC5AC levels than MVE group (P<0.05). Conclusions Exposure to CS or MVE can establish a rat model of COPD, with CS exposure better mimicking characteristics of acute exacerbation of COPD compared to MVE exposure.

Objective To analyze the effects and mechanisms of a disintegrin and metalloproteinase with thrombospondin motifs 14 (ADAMTS14) gene in gastric cancer associated with Helicobacter pylori (Hp) infection and explore the potential of ADAMTS14 as a novel target for biological therapy of gastric cancer. Methods The Caner Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were utilized to analyze differentially expressed genes in gastric cancer tissues and Hp-positive gastric mucosa, and to screen potential targets. For the selected target ADAMTS14, its expression pattern in gastric cancer and Hp-positive gastric mucosa was analyzed. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was performed on the genes co-expressed with ADAMTS14 in gastric cancer. Kaplan-Meier Plotter database was used to analyze the correlation between high- and low-ADAMTS14 expression and prognosis of gastric cancer patients. The expression of ADAMTS14 and its relationship with clinicopathological features in 30 patients with gastric cancer were analyzed using real-time fluorescent quantitative PCR (RT-qPCR) and immunohistochemistry. The relationship between Hp infection and ADAMTS14 expression was analyzed using GEO database, and confirmed by cell line infection experiment. ADAMTS14 expression in gastric cancer cell line NCI-N87 was silenced by small interfering RNA to analyze its effect on the function of gastric cancer cells; The effect of ADAMTS14 knockdown on downstream target genes of Hippo signaling pathway was analyzed by RT-qPCR and Western blotting. Results A total of 16 genes closely related to Hp infection and gastric cancer were identified by analyzing the TCGA and GEO databases. According to TCGA database, the ADAMTS14 expression in gastric cancer tissues was higher than that in healthy gastric mucosa (P<0.0001). RT-qPCR and immunohistochemistry results also showed higher ADAMTS14 expression in gastric cancer tissues compared to adjacent tissues (P<0.01). Survival analysis demonstrated poor prognosis in patients with high ADAMTS14 expression (P<0.001). Data from GSE60427 showed that ADAMTS14 expression was upregulated in Hp-infected gastric mucosa and cell lines (P<0.001). After Hp P12 infection, the ADAMTS14 expression in NCI-N87 cells was higher than that in uninfected group. ADAMTS14 knockdown significantly inhibited the proliferation and migration of NCI-N87 cells (P<0.05), and it also significantly inhibited the expression of Hippo signaling pathway target genes, including BMP7, BMPR2, FZD4, PARD3 and SAV1 (P<0.05). Conclusion ADAMTS14 is highly expressed in Hp-positive gastric mucosa and gastric cancer tissues, which may accelerate gastric cancer progression by regulating Hippo signaling pathway, and holds potential as a new marker and therapeutic target for Hp infective gastric cancer.

Objective To evaluate the methodological and reporting quality of published guidelines/consensus for childhood autism (CA), providing a basis for formulating domestic CA guidelines. Methods We searched databases including CNKI, Wanfang Data, SinoMed, Medlive, PubMed, national institute of health and clinical excellence (NICE), national guideline clearinghouse (NGC), and Scottish intercollegiate guidelines network (SIGN) for Chinese and foreign guideline/consensus on childhood autism published before February 1, 2024. Two researchers independently evaluated the methodology and reporting quality of the guideline/consensus using the Appraisal of Guidelines Research and Evaluation Ⅱ (AGREE Ⅱ) and Reporting Items for Practice Guidelines in Healthcare (RIGHT) tools. Results After literature screening, 19 CA guidelines/consensus were included, comprising 11 guidelines, 7 consensus, and 1 expert recommendation, with 9 domestic and 10 foreign articles. The AGREE Ⅱ evaluation scores for the six domains were as follows: scope and purpose (91.1%±4.5%), stakeholder involvement (86.8%±6.7%), rigour of development (83.0%±10.2%), clarity of presentation (84.3%±6.2%), applicability (82.7%±13.3%), and editorial independence (65.4%±21.8%). The RIGHT checklist reported rates for the seven domains were: basic information (87.6%±11.0%), background (87.6%±13.8%), evidence (81.1%±22.6%), recommendation (71.1%±38.4%), review and quality assurance (83.5%±16.7%), funding and declaration and management of interests (48.7%±29.4%), and other information (64.4%±11.8%). The domain with the lowest score for methodological quality was "editorial independence" and for reporting quality, it was "funding and declaration and management of interests". The reporting rate of domestic articles (26.2%±1.5%) was significantly lower than that of foreign articles (52.6%±2.2%), with a statistically significant difference (P<0.05). Conclusion The overall quality of current childhood autism guidelines/consensus requires improvement. During the formulation and reporting of guidelines/consensus, strictly adhering to AGREE Ⅱ and RIGHT is imperative, and it is essential to clearly report funding sources and conflicts of interest.

Objective To analyze the clinical efficacy of total laparoscopic proximal gastrectomy and esophagogastric anastomosis to provide a new surgical method for the treatment of early proximal gastric cancer. Methods The clinical data of 80 patients with early gastric cancer who underwent proximal gastrectomy in the Department of General Surgery, the First Medical Center of Chinese PLA General Hospital from January 2019 to June 2021 were retrospectively analyzed. According to different surgical methods, the patients were divided into two groups: a total laparoscopic proximal gastrectomy group (n=36) and a laparoscopic-assisted proximal gastrectomy group (n=44). The perioperative conditions, long-term complications, and survival status were compared between the two groups. Results The length of surgical incision [(59.9±4.7) mm vs. (119.7±8.3) mm, P<0.001], first exhaust time [(58.2±15.3) h vs. (66.8±16.4) h, P=0.019] and postoperative hospital stay [(7.6±1.1) d vs. (9.2±1.3) d, P<0.001] of total laparoscopic proximal gastrectomy group were significantly shorter than those of laparoscope-assisted proximal gastrectomy group. The duration of operation [(186.9±16.4) min vs. (154.0±17.2) min, P<0.001] of total laparoscopic proximal gastrectomy group was significantly longer than that of laparoscopic-assisted proximal gastrectomy group. There were no significant differences in intra-operative hemorrhage, number of lymph nodedissection and first liquid diet feeding time between the two groups (P>0.05), and no early complications requiring surgical intervention occurred. The incidence of reflux esophagitis in the total laparoscopic proximal gastrectomy group was less than that in laparoscope-assisted proximal gastrectomy group [16.7%(6/36) vs. 38.6%(17/44), P=0.031]. There was no significant difference in gastric motility, residual gastritis, anastomosis stenosis and anastomotic ulcer between the two groups (P>0.05). Conclusion Total laparoscopic proximal gastrectomy and esophagogastric anastomosis have the characteristics of good safety, rapid postoperative recovery and anti-reflux, being worthy of clinical popularization.

Lipids, including fats (triglycerides) and lipoids (phospholipids and sterols), not only serve as an energy source for the body but also play a pivotal role throughout the reproductive process, particularly in the establishment and maintenance of early pregnancy. This encompasses the regulate of early embryonic development and uterine tolerance, and the facilitation of embryo implantation. Given the diversity of lipids, this review focuses on extensively studied lipid mediators such as polyunsaturated fatty acids, endocannabinoids, prostaglandins, lysophosphatidic acid, sphingolipids and steroid hormones. It systematically elaborates on the regulatory effects of fatty acid, phospholipid, and cholesterol metabolism on the formation of endometrial receptivity and embryo implantation, as well as the potential underlying mechanisms. The review aims to provide new insights and feasible intervention approaches for predicting and improving the outcomes of natural pregnancy and/or assisted reproductive technology.

Objective To investigate the effect of Mex3c gene knockout on embryonic neural tube development and its possible mechanisms. Methods The NCBI database was used to analyze the expression of Mex3c gene in various tissues of mice. Fluorescence in situ hybridization (FISH) was employed to detect the expression of Mex3c in neural tubes of Mex3c^+/+ mice at different developmental stages (E12.5 d, E14.5 d). Sexual mature mice were mated at a ratio of Mex3c^+/- male to female (1:1) in the same cage. Embryos were collected and genotyped using PCR. They were divided into 3 groups based on their genotype: wild-type group (Mex3c^+/+, WT group), homozygous knockout group (Mex3c^-/-, KO group), and heterozygous knockout group (Mex3c^+/-). HE staining was employed to observe the development of neural tubes in the 3 groups of embryos. Immunofluorescence staining and Western blotting were performed to detect the proliferation and apoptosis of embryonic neural stem cells in the WT and KO groups. Transmission electron microscopy was used to observe the ultrastructure of the neural tubes and mitochondria in the WT and KO groups. RNA was extracted from the neural tubes of WT and KO groups for RNA-seq sequencing. The R.3.6.3 software was used to perform KEGG signaling pathway enrichment analysis on differentially expressed genes. RT-qPCR was used to validate the sequencing results. Results The NCBI database analysis and FISH detection results showed that the Mex3c gene was mainly expressed in the central nervous system of embryos. HE staining results showed that there was no significant difference in the development of embryonic neural tubes between KO group, WT group, and heterozygous knockout group at E12.5 d and E13.5 d. However, at E14.5 d, the embryonic neural tube development in KO group was delayed and the phenotype was significantly abnormal compared with those in WT group. Therefore, the embryonic neural tube tissues of KO group and WT group at E14.5 d were selected for subsequent experiments. The immunofluorescence staining results showed that the PCNA positive cell rate in KO group was significantly lower than that in WT group (P<0.001). The Western blotting results showed that the Bax/Bcl-2 ratio in KO group was higher than that in WT group (P<0.01). Transmission electron microscopy observation showed that compared with WT group, the synaptic gap in KO group disappeared, the mitochondrial of the embryonic neural tube in KO group were swollen, the mitochondrial cristae were disrupted, and the structure was significantly abnormal. The results of RNA-seq analysis showed that a total of 377 differentially expressed genes were obtained, including 101 up-regulated genes and 276 down-regulated genes. KEGG signaling pathway enrichment analysis revealed that the main signaling pathways of differentially expressed genes were enriched in the neuroactive ligand receptor interaction signaling pathways. The RT-qPCR validation results showed that the mRNA expression levels of Avpr1a, Drd1, Htr7, Sstr1, Oxtr and Gabra5 in this signaling pathway were down-regulated (P<0.05 or P<0.01), which was consistent with the RNA-seq results. Conclusion Mex3c plays an important role in the development of neural tubes in mouse embryos, which may participate in regulating the proliferation and apoptosis of neural stem cells through neural active ligand receptor interaction signaling pathways, thereby affecting the development of neural tubes.