In situ tumor vaccine has become an important strategy in cancer immunotherapy owing to its ability to induce immune responses locally and overcome tumor heterogeneity. However, the abnormal structure and mechanical properties of the tumor’s physical microenvironment significantly limit the efficiency of vaccine delivery and immune efficacy. In this review, the key factors in the tumor’s physical microenvironment, including solid pressure, interstitial fluid pressure, matrix stiffness, and tissue microstructure, are systematically discussed. Their obstructive roles in immune cell infiltration, antigen presentation, and immune activation are analyzed. The potential of approaches, such as radiotherapy, anti-angiogenic therapy, extracellular matrix degradation agents, nanomaterials, and hydrogel delivery platforms, in reshaping the tumor’s physical microenvironment is explored. This review aims to offer theoretical and practical guidance for optimizing in situ vaccine strategies through the regulation of the tumor’s physical microenvironment, ultimately advancing the precision and effectiveness of cancer immunotherapy.

Objective:s To investigate the risk factors for delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) in patients with acute carbon monoxide poisoning (ACOP) and to develop predictive models based on machine learning algorithms.Methods:Patients with ACOP hospitalized at the First Affiliated Hospital of Zhengzhou University from August 2019 to October 2024 were included, with the occurrence of DEACMP as the outcome measure. The dataset was randomly divided into training and validation sets at a ratio of 7:3. Lasso regression was used to select features influencing the outcome in training sets. Nine machine learning models—including Random Forest (RF), Extreme Gradient Boosting (XGBoost), and Support Vector Machine (SVM)—were constructed. Receiver operating characteristic (ROC) curves were plotted and the area under the curve (AUC) calculated for each model. Calibration curves were used to assess accuracy, and decision curve analysis (DCA) was applied to evaluate clinical utility. The SHapley Additive exPlanations (SHAP) method was employed to visualize and interpret the best-performing model.Results:A total of 264 ACOP patients were included, of whom 54 (20.5%) developed DEACMP. Lasso regression identified eight key feature variables. Based on these factors, predictive models were constructed, showing good AUC stability across the nine machine learning models in both training (0.92–0.99) and validation sets (0.85–0.91). The RF model performed best, with an AUC of 0.99 in the training set and 0.90 in the validation set; its calibration curve and DCA curve also demonstrated excellent performance. SHAP analysis of the RF model revealed the importance ranking of factors from highest to lowest as follows: Glasgow Coma Scale (GCS) score, duration of coma, age, history of coronary heart disease, CK-MB level, monocyte count, diastolic blood pressure (DBP), and drinking history.Conclusions:The RF model exhibited the highest predictive performance for DEACMP occurrence in ACOP patients. The influencing factors, ranked in order of importance from highest to lowest, are as follows: GCS score, duration of coma, age, history of coronary heart disease, CK-MB level, monocyte count, DBP, and drinking history.

Objective: To explore the factors affecting language development delay among children aged <3 years, so as to provide a basis for the prevention and early intervention of children's language development problems. Methods: Eighty-one children aged <3 years with language development delay who visited the children's language development clinic of Shaoxing Maternal and Child Health Hospital from January to December 2024 as the case group. Meanwhile, 118 children who underwent routine physical examinations at the children's health clinic during the same period, had normal language development were randomly selected as the control group. Data on children's basic information, parenting environment, and screen exposure were collected through questionnaire surveys. Language development delay was assessed using the Early Language Milestone Scale and the Gesell Developmental Diagnosis Scale. The factors for language development delay were analyzed using a multivariable logistic regression model. Results: The case group comprised 81 children, including 56 boys (69.14%) and 25 girls (30.86%), with a mean age of (23.14±4.84) months. The control group consisted of 118 children, including 81 boys (68.64%) and 37 girls (31.36%), with a mean age of (23.81±4.60) months. Multivariable logistic regression analysis showed that daily parental companionship time of ≥2 hours (OR=0.121, 95%CI: 0.040-0.367), attending childcare institutions (OR=0.103, 95%CI: 0.030-0.352), the average daily screen exposure time <1 hour (OR=0.614, 95%CI: 0.400-0.942), interactive parental accompaniment during screen exposure (OR=0.350, 95%CI: 0.157-0.779), and restricting screen exposure time (OR=0.162, 95%CI: 0.056-0.470) were associated with a lower risk of language development delay among children aged <3 years. Conclusion Daily paternal companionship of 2 hours or more, attending childcare institutions, daily screen exposure time of less than 1 hour, interactive parental companionship during screen time, and limiting screen exposure time can reduce the risk of language developmental delay among children aged under 3 years.

Objective:To compare the clinical efficacy and adverse events of chemoradiotherapy (CRT) combined with immunotherapy versus CRT alone in patients with locally recurrent esophageal squamous cell carcinoma (ESCC) after surgery.Methods:A total of 221 patients with postoperative locally recurrent ESCC who underwent CRT at the Affiliated Huai′an No. 1 People′s Hospital of Nanjing Medical University from January 2020 to December 2022 were retrospectively analyzed. Patients were divided into two groups according to treatment modality: the CRT group ( n=118) and the CRT plus immunotherapy group (combined group, n=103). Among the combined group, 39 patients received camrelizumab, 33 received sintilimab, and 31 received tislelizumab. Short-term efficacy, survival outcomes, and treatment-related adverse events were compared between the two groups. The Cox proportional hazards model was used to analyze prognostic factors for overall survival (OS). Results:No statistically significant differences were observed in baseline clinicopathological characteristics between the two groups. The objective response rate (ORR) in the combined group was 72.8%, significantly higher than 55.9% in the CRT group ( P=0.009). The 1- and 2-year OS rates in the CRT group were 68.6% and 41.5%, respectively, while the 1- and 2-year progression-free survival (PFS) rates were 56.8% and 30.5%, respectively. In the combined group, the 1- and 2-year OS rates were 84.5% and 55.3%, and the 1- and 2-year PFS rates were 67.0% and 42.7%, respectively. The differences in both OS and PFS between the two groups were statistically significant ( P=0.001 and 0.023, respectively). Multivariate analysis showed that Karnofsky performance status score of 70, TNM stage III, and CRT alone were independent risk factors for OS ( P=0.035, 0.031, and 0.002, respectively). The incidence of grade ≥3 adverse events did not differ significantly between the two groups ( P=0.550). A total of 85 grade 1-2 immune-related adverse events (irAEs) and 7 grade ≥3 irAEs occurred in 65 patients in the combined group. Subgroup analysis suggested that tislelizumab combined with CRT showed the most favorable efficacy and safety profile. Conclusion:For patients with locally recurrent ESCC after surgery, concurrent chemoradiotherapy combined with immunotherapy demonstrates promising efficacy with tolerable safety, and may offer a potential therapeutic advantage.

Objective:To explore the changes of aquaporin-4 (AQP4) expression in rat astrocytes (RA) and rat model of radiation-induced brain injury (RIBI) after X-ray radiation, as well as its impact on cell functions.Methods:Logarithmic-phase RA were divided into four groups: Sham group (no treatment), AQP4 siRNA group (transfected with AQP4 lentivirus), IR group (single 20 Gy X-ray irradiation), and IR+AQP4 siRNA group (transfected with AQP4 lentivirus followed by single 20 Gy X-ray irradiation). The cell proliferation viability after radiation was detected using the CCK-8 assay. The relative expression of AQP4 mRNA in each group was measured by quantitative reverse transcription polymerase chain reaction, and the optimal AQP4 siRNA lentiviral sequence was selected for further studies. Western blot was used to detect the relative expression levels of AQP4, phosphorylated histone H2A family member X (γH2AX), autophagy-related proteins, glial fibrillary acidic protein (GFAP), proteins in the phosphatidylinositol-3-kinase (PI3K) / protein kinase B (AKT) / mammalian target of rapamycin (mTOR) pathway, and apoptosis-related proteins in cells from each group. Immunofluorescence (co-expression) staining was used to detect the expression of AQP4, γH2AX, GFAP, LC3, phosphatase and tensin homolog-induced kinase 1 (PINK1) in cells from each group, as well as the expression of AQP4 and LC3 in brain tissues from the rat RIBI model. Flow cytometry was used to detect the apoptosis rate of cells in each group. The differences between two groups were compared using the t-test, while the differences among multiple groups were evaluated using analysis of variance (ANOVA) and Tukey's multiple comparison test. Results:The proliferation viability of RA decreased to its lowest level at 24 h after irradiation. At the 24 h post-irradiation time point, the expression levels of γH2AX and AQP4 were significantly increased in the IR group compared with the Sham group. The fluorescence intensity of AQP4 in the AQP4 siRNA group was lower than that in the Sham group. The cells in the IR group showed significant enlargement and irregular morphology, with a significant increase in AQP4 fluorescence intensity, while the IR+AQP4 siRNA group showed mild cell enlargement and lower AQP4 fluorescence intensity compared with the IR group. Compared with the Sham group, the expression of microtubule-associated protein1 light chain 3 (LC3) and PINK1 proteins were decreased in the IR group, while the IR+AQP4 siRNA group showed higher expression levels compared with the IR group. Compared with the Sham group, the IR group showed increased expression of AQP4, GFAP, and sequestosome 1 (P62) proteins, decreased expression of Beclin-1 (BECN1) and LC3 proteins, and a reduced LC3-II/LC3-I ratio, while the IR+AQP4 siRNA group exhibited a restored LC3-II/LC3-I ratio. The apoptosis rate in the AQP4 siRNA group was similar to that in the Sham group, while the IR group showed a significantly increased apoptosis rate, and the IR+AQP4 siRNA group had a lower apoptosis rate than the IR group. The expression of caspase-3 (Caspase3) decreased in the IR group, while the expression of PI3K, AKT, mTOR, extracellular signal-regulated kinase (ERK)1/2, and cleaved Caspase3 protein increased; these changes were partially reversed in the IR+AQP4 siRNA group. In the rat RIBI model, the expression area and intensity of AQP4 were higher in the IR group compared with the Sham group, while LC3 expression showed the opposite pattern.Conclusions:The possible molecular mechanism of RA apoptosis caused by X-ray irradiation is that irradiation promotes the expression of AQP4, causes cell swelling, inhibits autophagy, and prevents cells from clearing damaged organelles in a timely manner, thereby promoting cell apoptosis.

Objective:To compare the clinical efficacy and adverse events of chemoradiotherapy (CRT) combined with immunotherapy versus CRT alone in patients with locally recurrent esophageal squamous cell carcinoma (ESCC) after surgery.Methods:A total of 221 patients with postoperative locally recurrent ESCC who underwent CRT at the Affiliated Huai′an No. 1 People′s Hospital of Nanjing Medical University from January 2020 to December 2022 were retrospectively analyzed. Patients were divided into two groups according to treatment modality: the CRT group ( n=118) and the CRT plus immunotherapy group (combined group, n=103). Among the combined group, 39 patients received camrelizumab, 33 received sintilimab, and 31 received tislelizumab. Short-term efficacy, survival outcomes, and treatment-related adverse events were compared between the two groups. The Cox proportional hazards model was used to analyze prognostic factors for overall survival (OS). Results:No statistically significant differences were observed in baseline clinicopathological characteristics between the two groups. The objective response rate (ORR) in the combined group was 72.8%, significantly higher than 55.9% in the CRT group ( P=0.009). The 1- and 2-year OS rates in the CRT group were 68.6% and 41.5%, respectively, while the 1- and 2-year progression-free survival (PFS) rates were 56.8% and 30.5%, respectively. In the combined group, the 1- and 2-year OS rates were 84.5% and 55.3%, and the 1- and 2-year PFS rates were 67.0% and 42.7%, respectively. The differences in both OS and PFS between the two groups were statistically significant ( P=0.001 and 0.023, respectively). Multivariate analysis showed that Karnofsky performance status score of 70, TNM stage III, and CRT alone were independent risk factors for OS ( P=0.035, 0.031, and 0.002, respectively). The incidence of grade ≥3 adverse events did not differ significantly between the two groups ( P=0.550). A total of 85 grade 1-2 immune-related adverse events (irAEs) and 7 grade ≥3 irAEs occurred in 65 patients in the combined group. Subgroup analysis suggested that tislelizumab combined with CRT showed the most favorable efficacy and safety profile. Conclusion:For patients with locally recurrent ESCC after surgery, concurrent chemoradiotherapy combined with immunotherapy demonstrates promising efficacy with tolerable safety, and may offer a potential therapeutic advantage.

Objective:To explore the changes of aquaporin-4 (AQP4) expression in rat astrocytes (RA) and rat model of radiation-induced brain injury (RIBI) after X-ray radiation, as well as its impact on cell functions.Methods:Logarithmic-phase RA were divided into four groups: Sham group (no treatment), AQP4 siRNA group (transfected with AQP4 lentivirus), IR group (single 20 Gy X-ray irradiation), and IR+AQP4 siRNA group (transfected with AQP4 lentivirus followed by single 20 Gy X-ray irradiation). The cell proliferation viability after radiation was detected using the CCK-8 assay. The relative expression of AQP4 mRNA in each group was measured by quantitative reverse transcription polymerase chain reaction, and the optimal AQP4 siRNA lentiviral sequence was selected for further studies. Western blot was used to detect the relative expression levels of AQP4, phosphorylated histone H2A family member X (γH2AX), autophagy-related proteins, glial fibrillary acidic protein (GFAP), proteins in the phosphatidylinositol-3-kinase (PI3K) / protein kinase B (AKT) / mammalian target of rapamycin (mTOR) pathway, and apoptosis-related proteins in cells from each group. Immunofluorescence (co-expression) staining was used to detect the expression of AQP4, γH2AX, GFAP, LC3, phosphatase and tensin homolog-induced kinase 1 (PINK1) in cells from each group, as well as the expression of AQP4 and LC3 in brain tissues from the rat RIBI model. Flow cytometry was used to detect the apoptosis rate of cells in each group. The differences between two groups were compared using the t-test, while the differences among multiple groups were evaluated using analysis of variance (ANOVA) and Tukey's multiple comparison test. Results:The proliferation viability of RA decreased to its lowest level at 24 h after irradiation. At the 24 h post-irradiation time point, the expression levels of γH2AX and AQP4 were significantly increased in the IR group compared with the Sham group. The fluorescence intensity of AQP4 in the AQP4 siRNA group was lower than that in the Sham group. The cells in the IR group showed significant enlargement and irregular morphology, with a significant increase in AQP4 fluorescence intensity, while the IR+AQP4 siRNA group showed mild cell enlargement and lower AQP4 fluorescence intensity compared with the IR group. Compared with the Sham group, the expression of microtubule-associated protein1 light chain 3 (LC3) and PINK1 proteins were decreased in the IR group, while the IR+AQP4 siRNA group showed higher expression levels compared with the IR group. Compared with the Sham group, the IR group showed increased expression of AQP4, GFAP, and sequestosome 1 (P62) proteins, decreased expression of Beclin-1 (BECN1) and LC3 proteins, and a reduced LC3-II/LC3-I ratio, while the IR+AQP4 siRNA group exhibited a restored LC3-II/LC3-I ratio. The apoptosis rate in the AQP4 siRNA group was similar to that in the Sham group, while the IR group showed a significantly increased apoptosis rate, and the IR+AQP4 siRNA group had a lower apoptosis rate than the IR group. The expression of caspase-3 (Caspase3) decreased in the IR group, while the expression of PI3K, AKT, mTOR, extracellular signal-regulated kinase (ERK)1/2, and cleaved Caspase3 protein increased; these changes were partially reversed in the IR+AQP4 siRNA group. In the rat RIBI model, the expression area and intensity of AQP4 were higher in the IR group compared with the Sham group, while LC3 expression showed the opposite pattern.Conclusions:The possible molecular mechanism of RA apoptosis caused by X-ray irradiation is that irradiation promotes the expression of AQP4, causes cell swelling, inhibits autophagy, and prevents cells from clearing damaged organelles in a timely manner, thereby promoting cell apoptosis.