Objective:To investigate the effect of PIK3CA mutations on the tumor immune microenvironment in Luminal breast cancer and evaluate the potential of Alpelisib-loaded pH-responsive polymer micelles in modulating the tumor immune microenvironment.Methods:PIK3CA mutations in breast cancer were analyzed using bioinformatics tools.A mouse xenograft model of Luminal breast cancer harboring a PIK3CA mutation was established,and alterations in the tumor immune microenvironment were examined using mass cytometry(CyTOF).During the period from August 2004 to December 2008 in Tianjin Medical University Cancer Hospital,tissue biopsies of 62 Luminal breast cancer patients in the BRCA cohort were collected Study the relationship between PIK3CA mutations and tumor immune microenvironment at the organizational level.Alpelisib-loaded polymer micelles(Alpelisib@MSPM)were synthesized,characterized,and evaluated for thera-peutic efficacy in Luminal breast cancer with PIK3CA mutations.Results:PIK3CA is one of the most frequently mutated genes in breast can-cer,with the highest prevalence in Luminal subtypes.CyTOF analysis demonstrated that PIK3CA mutations contribute to a tumor immun-osuppressive microenvironment in xenografts.Multiplex fluorescence immunohistochemistry revealed that PIK3CA-mutated tumors exhib-ited more infiltration of myeloid-derived suppressor cells(MDSCs)and less infiltration of CD8? T cells.The synthesized Alpelisib-loaded pH-re-sponsive polymer micelles had an average size of approximately 127 nm.Treatment with Alpelisib and Alpelisib@MSPM reduced tumor growth in mice with PIK3CA-mutated Luminal breast cancer.Notably,the proportion of MDSCs decreased,whereas CD8? T cell infiltration in-creased significantly,with the more pronounced effect observed in the Alpelisib@MSPM treatment group.Conclusions:PIK3CA mutations drive the formation of a tumor immunosuppressive microenvironment in Luminal breast cancer.Targeted Alpelisib delivery via pH-respons-ive polymer micelles significantly enhances therapeutic efficacy in PIK3CA-mutated breast cancer.

Objective:To develop and validate clinical predictive models for identifying poor short-term response to recombinant human growth hormone(rhGH) treatment in children with short stature.Methods:A retrospective analysis was conducted on 118 children diagnosed with growth hormone deficiency or idiopathic short stature who were treated at the First Affiliated Hospital of Zhengzhou University and two other hospitals between January 1, 2020, and January 1, 2024. A poor response to rhGH was defined as a height increase of less than 0.2 standard deviation score(SDS) after 6 months of rhGH treatment. LASSO regression was used to identify predictive variables from baseline and follow-up data. Two logistic regression models were conducted: Model A(incorporating baseline variables only) and model B(incorporating both baseline and follow-up variables), and nomograms were created for visualization. External data and internal resampling were used for dual validation of the models, and their performance was compared.Results:A total of 118 children with short stature were included. Six baseline predictive variables(diagnosis, initial height SDS, bone age, bone age-chronological age difference, rhGH dose, and gender) and one follow-up variable(height SDS after 3 months of rhGH treatment) were identified. Area under the curve values for Model A and Model B were 0.753(95% CI 0.696-0.811) and 0.930(95% CI 0.891-0.975), respectively. Calibration curves, decision curve analysis, and other evaluation metrics demonstrated good discrimination and clinical utility for both models. Model B, incorporating the 3-month follow-up variable, showed superior predictive performance compared to Model A. Conclusions:The clinical prediction models developed in this study(Model A and Model B) are practical and reliable tools for quantitatively, conveniently, and intuitively identifying children with short stature at risk of poor response to rhGH treatment.

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

Objective:To summarize the clinical and genetic characteristics of 23 children with pathogenic ACAN gene variants, enhance the understanding of this disorder, and explore possible regulatory mechanisms. Methods:A retrospective case series summary.The clinical characteristics and genetic analysis results of 23 children with ACAN gene variants treated in the Department of Endocrinology, Genetics and Metabolism, Children′s Hospital of Soochow University from January 2016 and September 2024 were analyzed retrospectively.Transcriptome sequencing was performed on peripheral blood samples from 3 of affected children and 3 age-matched healthy children as controls.Differentially expressed genes (DEGs) in the peripheral blood transcriptome profiles were identified.Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses were conducted to explore the potential signaling pathways involved. Results:Among the 23 cases, there were 13 males and 10 females, aged from 2 years and 8 months to 12 years old, with 11 cases presenting advanced bone age.Thirteen cases were treated with growth hormone (GH), including 6 cases who received concomitant gonadotropin-releasing hormone analogue therapy.The treatment duration ranged from 3 to 70 months, resulting in varying degrees of height improvement in all treated patients.Transcriptomic analysis identified 811 DEGs, with 516 up-regulated and 295 down-regulated.GO and KEGG enrichment analyses revealed that the heterozygous ACAN variants were significantly associated with FcγR-mediated phagocytosis, nuclear factor-κB signaling pathway, the intestinal immune network for IgA production, rheumatoid arthritis, and systemic lupus erythematosus signaling pathways. Conclusions:The predominant clinical manifestations of patients with ACAN gene variants are short stature and advanced bone age.Although GH provocation tests may indicate normal GH levels, GH therapy can be effective in improving height.Immune-related factors may play a role in the pathogenesis of this disorder.

Objective:To summarize the clinical and genetic characteristics of 23 children with pathogenic ACAN gene variants, enhance the understanding of this disorder, and explore possible regulatory mechanisms. Methods:A retrospective case series summary.The clinical characteristics and genetic analysis results of 23 children with ACAN gene variants treated in the Department of Endocrinology, Genetics and Metabolism, Children′s Hospital of Soochow University from January 2016 and September 2024 were analyzed retrospectively.Transcriptome sequencing was performed on peripheral blood samples from 3 of affected children and 3 age-matched healthy children as controls.Differentially expressed genes (DEGs) in the peripheral blood transcriptome profiles were identified.Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses were conducted to explore the potential signaling pathways involved. Results:Among the 23 cases, there were 13 males and 10 females, aged from 2 years and 8 months to 12 years old, with 11 cases presenting advanced bone age.Thirteen cases were treated with growth hormone (GH), including 6 cases who received concomitant gonadotropin-releasing hormone analogue therapy.The treatment duration ranged from 3 to 70 months, resulting in varying degrees of height improvement in all treated patients.Transcriptomic analysis identified 811 DEGs, with 516 up-regulated and 295 down-regulated.GO and KEGG enrichment analyses revealed that the heterozygous ACAN variants were significantly associated with FcγR-mediated phagocytosis, nuclear factor-κB signaling pathway, the intestinal immune network for IgA production, rheumatoid arthritis, and systemic lupus erythematosus signaling pathways. Conclusions:The predominant clinical manifestations of patients with ACAN gene variants are short stature and advanced bone age.Although GH provocation tests may indicate normal GH levels, GH therapy can be effective in improving height.Immune-related factors may play a role in the pathogenesis of this disorder.

Objective:To investigate the effect of PIK3CA mutations on the tumor immune microenvironment in Luminal breast cancer and evaluate the potential of Alpelisib-loaded pH-responsive polymer micelles in modulating the tumor immune microenvironment.Methods:PIK3CA mutations in breast cancer were analyzed using bioinformatics tools.A mouse xenograft model of Luminal breast cancer harboring a PIK3CA mutation was established,and alterations in the tumor immune microenvironment were examined using mass cytometry(CyTOF).During the period from August 2004 to December 2008 in Tianjin Medical University Cancer Hospital,tissue biopsies of 62 Luminal breast cancer patients in the BRCA cohort were collected Study the relationship between PIK3CA mutations and tumor immune microenvironment at the organizational level.Alpelisib-loaded polymer micelles(Alpelisib@MSPM)were synthesized,characterized,and evaluated for thera-peutic efficacy in Luminal breast cancer with PIK3CA mutations.Results:PIK3CA is one of the most frequently mutated genes in breast can-cer,with the highest prevalence in Luminal subtypes.CyTOF analysis demonstrated that PIK3CA mutations contribute to a tumor immun-osuppressive microenvironment in xenografts.Multiplex fluorescence immunohistochemistry revealed that PIK3CA-mutated tumors exhib-ited more infiltration of myeloid-derived suppressor cells(MDSCs)and less infiltration of CD8? T cells.The synthesized Alpelisib-loaded pH-re-sponsive polymer micelles had an average size of approximately 127 nm.Treatment with Alpelisib and Alpelisib@MSPM reduced tumor growth in mice with PIK3CA-mutated Luminal breast cancer.Notably,the proportion of MDSCs decreased,whereas CD8? T cell infiltration in-creased significantly,with the more pronounced effect observed in the Alpelisib@MSPM treatment group.Conclusions:PIK3CA mutations drive the formation of a tumor immunosuppressive microenvironment in Luminal breast cancer.Targeted Alpelisib delivery via pH-respons-ive polymer micelles significantly enhances therapeutic efficacy in PIK3CA-mutated breast cancer.

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

Objective:To develop and validate clinical predictive models for identifying poor short-term response to recombinant human growth hormone(rhGH) treatment in children with short stature.Methods:A retrospective analysis was conducted on 118 children diagnosed with growth hormone deficiency or idiopathic short stature who were treated at the First Affiliated Hospital of Zhengzhou University and two other hospitals between January 1, 2020, and January 1, 2024. A poor response to rhGH was defined as a height increase of less than 0.2 standard deviation score(SDS) after 6 months of rhGH treatment. LASSO regression was used to identify predictive variables from baseline and follow-up data. Two logistic regression models were conducted: Model A(incorporating baseline variables only) and model B(incorporating both baseline and follow-up variables), and nomograms were created for visualization. External data and internal resampling were used for dual validation of the models, and their performance was compared.Results:A total of 118 children with short stature were included. Six baseline predictive variables(diagnosis, initial height SDS, bone age, bone age-chronological age difference, rhGH dose, and gender) and one follow-up variable(height SDS after 3 months of rhGH treatment) were identified. Area under the curve values for Model A and Model B were 0.753(95% CI 0.696-0.811) and 0.930(95% CI 0.891-0.975), respectively. Calibration curves, decision curve analysis, and other evaluation metrics demonstrated good discrimination and clinical utility for both models. Model B, incorporating the 3-month follow-up variable, showed superior predictive performance compared to Model A. Conclusions:The clinical prediction models developed in this study(Model A and Model B) are practical and reliable tools for quantitatively, conveniently, and intuitively identifying children with short stature at risk of poor response to rhGH treatment.

Health insurance fund regulation (HIFR) is a vital issue in the modernization of healthcare security governance, with its importance as a primary task of the healthcare security department continually reinforced in policy practice. This study focused on the 22 specialized policies issued by the National Healthcare Security Administration from its establishment in 2018 to March 2024, deeply analyzed their issue characteristics, and summarized the evolutionary trends of policy changes, as well as the knowledge production patterns that existed in the series of policy formulation, implementation, and feedback processes. Our analysis revealed that the diverse issue characteristics had led to heterogeneous directions in HIFR policies. The policy development process presented distinct composite evolutionary trends, mainly manifested in four aspects: the integration of regulatory system and content, the convergence of professional and societal forces, the parallelism of special governance and regular supervision, and the complementarity of conventional and emerging methods. Additionally, the study demonstrated that the knowledge production embedded in policy evolution encompassed four different dimensions: problem rectification, norm setting, pilot experience, and technical absorption. Together with issue characteristics and policy evolution, they formed an integrated, dynamic, and open system of knowledge production, continuously promoting the renewal and iteration of regulation policies.

Objective:To explore the clinical characteristics and prognostic influencing factors of multiple primary lung cancer (MPLC), and to construct a prognostic prediction model.Methods:The clinical data and prognostic information of MPLC patients diagnosed by pathological examination included in the Surveillance, Epidemiology, and End Results (SEER) database from January 2010 to December 2020 were retrospectively analyzed. Patients were randomly divided into training and validation sets according to a 7:3 ratio using R software. Survival curves were plotted by using the Kaplan-Meier method and log-rank test was used for comparison between groups. The independent influencing factors of overall survival (OS) of MPLC patients in the training set were screened using univariate and multivariate Cox proportional hazards models, and accordingly, the nomogram predicting the survival rate of patients at 3, 5 and 8 years were plotted. In the training and validation sets, using the actual survival as the gold standard, the receiver operating characteristic (ROC) curves of the constructed models for predicting the patients' 3-, 5- and 8-year OS rates were plotted, the area under the curve (AUC) was obtained, and C-index of the model was analyzed by using R software. The calibration curves of 3-, 5- and 8-year OS rates predicted by the models and the actual OS rates were plotted.Results:A total of 5 495 MPLC patients were included, 3 846 in the training set and 1 649 in the validation set. The differences in the composition of patients of different ages and AJCC stages between the training and validation sets were statistically significant (both P < 0.05), and the differences in the comparison of other clinicopathological characteristics were not statistically significant (all P > 0.05). The results of multivariate Cox regression analysis showed that males (compared with females, HR = 1.256, 95% CI: 1.144-1.379, P < 0.001), age ≥ 70 years old (compared with 50-59 years old, HR = 1.201, 95% CI: 1.030-1.400, P = 0.019), FPLC with pathological types of squamous cell carcinoma or other types (compared with adenocarcinoma, HR = 1.275, 95% CI: 1.137-1.431, P < 0.001; HR = 1.208, 95% CI: 1.041-1.403, P = 0.013), and SPLC with pathological types of squamous cell carcinoma, small cell lung carcinoma, or other types (compared with adenocarcinoma, HR = 1.270, 95% CI: 1.121-1.440, P < 0.001; HR = 1.978, 95% CI: 1.642-2.384, P < 0.001; HR = 1.246, 95% CI: 1.090-1.424, P = 0.001), and AJCC stage Ⅲ and Ⅳ (compared with stage Ⅰ, HR = 1.645, 95% CI: 1.447-1.869, P < 0.001; HR = 2.078, 95% CI: 1.669-2.587, P < 0.001), FPLC without operation (compared with operation, HR = 1.263, 95% CI: 1.038-1.536, P = 0.020), SPLC without operation (operation vs. no operation, HR = 0.680, 95% CI: 0.579-0.799, P < 0.001), FPLC without lymph node dissection or with clearance of 1-3 regional lymph nodes (compared with clearance of ≥4, HR = 1.225, 95% CI: 1.016-1.477, P = 0.034; HR = 1.314, 95% CI: 1.103-1.566, P = 0.002), FPLC with maximum diameter 3-5 cm or >5 cm (compared with <3 cm, HR = 1.181, 95% CI: 1.053-1.324, P = 0.005; HR = 1.232, 95% CI: 1.069-1.420, P = 0.004), and SPLC with maximum diameter 3-5 cm or >5 cm (compared with <3 cm, HR = 1.560, 95% CI: 1.362-1.786, P < 0.001; HR = 1.727, 95% CI: 1.451-2.054, P < 0.001), and FPLC without chemotherapy (chemotherapy vs. no chemotherapy or unknown, HR = 0.744, 95% CI: 0.655-0.845, P < 0.001) were the independent risk factors of patients' poor OS (all P < 0.05). The results of Kaplan-Meier survival analysis showed that the OS of patients with different gender, race, age, two tumor locations, AJCC staging, pathological type of two lung tumors, maximum diameter of two tumors, and whether two tumors were treated surgically or not, and whether two tumors were treated with chemotherapy or not in the training set were compared, and the differences were all statistically significant (all P < 0.05). Based on the independent factors affecting the OS of MPLC patients screened by the results of multivariate Cox regression analysis, nomogram predicting the 3-, 5- and 8-year OS rates of MPLC were plotted. The results of ROC curve analysis showed that the C-index of the training set's nomogram was 0.679 (95% CI: 0.649-0.701), and the AUC values for predicting the 3-, 5- and 8-year OS rates were 0.601, 0.595 and 0.586, respectively; the C-index of the validation set was 0.678 (95% CI: 0.633-0.720), and the AUC values for predicting 3-, 5- and 8-year OS rates were 0.643, 0.631 and 0.626, respectively. The calibration curves showed that the 3-, 5- and 8-year OS rates of patients predicted by the nomogram models in both the training and validation sets were in good agreement with the actual results with a high goodness-of-fit. Conclusions:The established prognostic model has good predictive value and can effectively assess the prognosis of patients.