ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

Objective To establish a sandwich enzyme-linked immunosorbent assay (ELISA) method for the quantitative detection of Chromogranin A (CHGA) in saliva, and to explore its preliminary application in the testing of saliva samples. Methods Recombinant human CHGA protein was used to immunize BALB/c mice, and monoclonal antibodies (mAbs) were prepared and screened using conventional hybridoma technology. A double-antibody sandwich ELISA detection method was constructed, and the matrix effect of saliva samples was optimized. This method was then applied to detect the concentration of CHGA in the saliva of stressed individuals. Results Twenty-one stable hybridoma cell lines secreting high affinity anti-human CHGA antibodies were obtained. A pair of detection antibodies with the best effect was selected, and the optimal coating concentration was determined to be 10 μg/mL, with the optimal dilution of detection antibodies being 1:32 000. The accuracy and reproducibility of this method were verified, with both intra-batch and inter-batch variation coefficients less than 15×, and the recovery rate between 80× and 120×. The matrix effect was further optimized to make it suitable for saliva sample detection. Saliva samples from individuals in different stress states were collected, and the CHGA levels were detected using the method established in this study, indicating its potential to reflect the intensity of stress. Conclusion A reliable saliva CHGA ELISA detection method has been successfully established, and its potential as a biomarker in stress-related research has been preliminarily explored.
Saliva/metabolism* ; Enzyme-Linked Immunosorbent Assay/methods* ; Humans ; Animals ; Mice, Inbred BALB C ; Mice ; Chromogranin A/immunology* ; Antibodies, Monoclonal/immunology* ; Female ; Male ; Reproducibility of Results ; Adult

Advances in targeted therapy and immunotherapy have significantly improved clinical outcomes for patients with advanced non-small cell lung cancer (NSCLC), reshaping treatment paradigms. However, most patients ultimately face drug resistance, with limited options for subsequent therapies and suboptimal treatment efficacy, presenting a prominent challenge in current clinical practice. Antibody-drug conjugates (ADCs), characterized by high efficacy and favorable safety profiles, have emerged as a promising therapeutic frontier in recent years. This systematic review provides a comprehensive overview of the latest advancements in ADCs-based therapies for lung cancer, alongside discussions of the prevailing challenges in this rapidly evolving domain.
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Humans ; Carcinoma, Non-Small-Cell Lung/immunology* ; Immunoconjugates/therapeutic use* ; Lung Neoplasms/immunology* ; Antineoplastic Agents/therapeutic use* ; Immunotherapy

Mutations in ion channel genes have long been implicated in a spectrum of epilepsy syndromes. However, therapeutic decision-making is relatively complex for epilepsies associated with channelopathy. Therefore, in the present study, we used a patient-derived organoid model with a novel SCN2A mutation (p.E512K) to investigate the potential of utilizing such a model as a platform for preclinical testing of anti-seizure compounds. The electrophysiological properties of the variant Nav1.2 exhibited gain-of-function effects with increased current amplitude and premature activation. Immunofluorescence staining of patient-derived cortical organoids (COs) displayed normal neurodevelopment. Multielectrode array (MEA) recordings of patient-derived COs showed hyperexcitability with increased spiking and remarkable network bursts. Moreover, the application of patient-derived COs for preclinical drug testing using the MEA showed that they exhibit differential responses to various anti-seizure drugs and respond well to carbamazepine. Our results demonstrate that the individualized organoids have the potential to serve as a platform for preclinical pharmacological assessment.
Organoids/physiology* ; NAV1.2 Voltage-Gated Sodium Channel/genetics* ; Humans ; Anticonvulsants/pharmacology* ; Epilepsy/drug therapy* ; Mutation ; Cerebral Cortex/drug effects* ; Action Potentials/drug effects* ; Carbamazepine/pharmacology*

Objective:This study utilizes data from the 2021 Chinese Longitudinal Healthy Longevity and Happy Family Survey(CLHLS-HF)to examine the current status of Long-Term Care Insurance(LTCI)implementation and to identify the factors influencing whether disabled elderly individuals receive formal care services.The study aims to provide policy recommendations to enhance the effectiveness and equity of the system.Methods:In this cross-sectional study, a sample of 1 447 older participants with dependency, residing in LTCI pilot areas and meeting the inclusion criteria from the 2021 CLHLS-HF, was selected.Chi-square tests and binary logistic regression analyses were employed to explore the factors influencing the receipt of formal care by dependent older individuals.Results:Among the 1 447 participants, there were 496 males with an average age of 92 years(SD 9)and 951 females with an average age of 95 years(SD 9). Of these, 701 received formal care.The logistic regression analysis revealed that factors influencing the receipt of formal care included urban residence( OR=2.237, 95% CI: 1.675-2.987, P<0.001), residing in the eastern region( OR=2.907, 95% CI: 1.747-4.837, P<0.001), living in the western region( OR=3.132, 95% CI: 1.816-5.501, P<0.001), having no children( OR=2.478, 95% CI: 1.108-5.540, P=0.027), and the degree of disability, with severe disability being more likely to receive care compared to mild( OR=0.497, 95% CI: 0.388-0.637, P<0.001)and moderate disabilities( OR=0.589, 95% CI: 0.433-0.801, P=0.001). Conclusions:Dependent older individuals in the eastern and western regions, particularly those without children or with severe disabilities, are more likely to receive formal care through the LTCI system.However, there are substantial inequities in LTCI coverage among individuals with varying degrees of disability.To enhance the effectiveness of the LTCI system, greater efforts should be directed towards economically disadvantaged regions and older individuals with mild to moderate disabilities, thereby ensuring better protection for the disabled population.

Objective:To analyze the trend of the health literacy level of Chinese residents from 2012 to 2023 and predict the health literacy level from 2024 to 2027.Methods:The study collected data on the health literacy surveillance of Chinese residents from 2012 to 2023. The Joinpoint regression model was used to calculate the average annual percent change (AAPC) and analyze the trend. The interrupted time series analysis with Prais-Winsten transformed generalized least squares estimation was employed to investigate the impact of the"Healthy China 2030" policy on residents′ health literacy levels. Joinpoint regression, autoregressive integrated moving average model and grey forecasting models were established to select the optimal model for forecasting health literacy levels from 2024 to 2027.Results:The results showed that the health literacy level of Chinese residents increased from 8.80% in 2012 to 29.70% in 2023 (AAPC=11.65%, P<0.05). The health literacy level of urban and rural residents increased from 11.79% and 7.13% in 2012 to 33.25% and 26.23% in 2023, respectively (AAPC=9.57% and 12.60%, both P<0.05). Rural (1.59% per year) saw a lower average annual increase than urban (1.79% per year), widening the urban-rural health literacy gap. All aspects of health literacy, including basic knowledge and concepts, healthy lifestyles and behaviors, and health skills, showed an upward trend. The literacy level of six health issues—safety and first aid, scientific health views, health information, infectious disease prevention, chronic disease prevention, and basic medical care—also exhibited rising trends. Interrupted time series analysis indicated a significant further increase in the health literacy level of Chinese residents after the implementation of the "Healthy China 2030" policy, with the growth rate increasing from 0.615% per year before implementation to 2.655% per year afterwards. The Joinpoint regression model showed superior predictive performance compared to autoregressive integrated moving average model and grey forecasting models. The prediction results suggested a continued upward trend in the health literacy level from 2024 to 2027, reaching 32.68%, 35.62%, 38.84%, and 42.34%, respectively. Conclusion:From 2012 to 2023, the overall and various aspects of health literacy among Chinese residents show a continuous upward trend. This study predicts that the level of residents′ health literacy will continue to rise by 2027.

Objective To compare the diagnostic performance of three breast MRI measurement methods—RECIST 1.1,the optimal method,and three-dimensional(3D)volumetric assessment—in assessing the efficacy of neoadjuvant chemotherapy(NAC)in breast cancer patients,with the objective of identifying the most clinically practical approach.Methods A total of 110 breast cancer patients who underwent NAC followed by surgical treatment between 2019 and 2023 were included in the study.Breast magnetic resonance imaging(MRI)was conducted within one week before and after the completion of NAC.Tumor response was evaluated using RECIST 1.1 criteria,widely recognized as the optimal method,as well as 3D volume measurement.Pathological response was determined according to the Miller-Payne grading system.Sensitivity,specificity,accuracy,and the area under the receiver operating characteristic curve(AUC)were computed and compared using the DeLong test.Results The AUC values for RECIST 1.1,the optimal method,and 3D volumetric assessment were 0.768,0.795,and 0.883,respectively.The 3D volumetric assessment exhibited significantly better discriminative performance(P＜0.05),with the highest sensitivity(98.9％),specificity(77.8％),and accuracy(95.5％).Additionally,the optimal method demonstrated superior performance over RECIST 1.1 across multiple parameters.Conclusions 3D volumetric mea-surement demonstrates superior performance compared to RECIST 1.1 and the optimal method in evaluating the response to NAC,offering a more accurate and comprehensive assessment tool.Additionally,the optimal method shows advantages over RECIST 1.1 and may serve as a practical alternative in settings where 3D software is not available.

Objective To analyze the characteristics and drug resistance of pathogenic bacteria isolated from patients undergoing solid organ transplantation(SOT)at West China Hospital,Sichuan University in re-cent years,in order to provide a basis for empirical anti infective treatment after SOT surgery.Methods A retrospective analysis was conducted on the isolation of major pathogens and their resistance to common anti-biotics from various specimens collected from patients undergoing kidney transplantation(KT),liver trans-plantation(LTx),and lung transplantation(LT)at West China Hospital,Sichuan University from 2017 to 2022.Results A total of 1 077 non-duplicate strains of pathogens were isolated from the samples of patients with infections after SOT surgery during the 6-year period,of which approximately 74.8％(806/1 077)were Gram negative bacteria and 25.2％(271/1 077)were Gram positive bacteria.There were differences in the distribution of pathogenic bacteria among different types of SOT groups and different specimens.Compared to E.coli isolated from urine specimens,the strains of E.coli isolated from non-urinary specimens exhibited a higher resistance rate to common antimicrobial drugs(P＜0.05).The resistance rate of E.coli to β-lactam/β-lactamase inhibitor combinations(cefoperazone/sulbactam and piperacillin/tazobactam)in the LTx group was significantly higher than that in the KT group(P＜0.05).The overall proportion of multidrug-resistant bac-teria after SOT surgery was 11.3％(122/1 077).The proportion of carbapenem resistant Acinetobacter bau-mannii and carbapenem resistant Pseudomonas aeruginosa among the same group and type of pathogens in the LTx group(93.8％,37.5％)was significantly higher than that in the KT group(55.8％,9.2％).Conclusion The specimen types and strain distribution of pathogenic bacteria after different types of SOT surgery are different.The same pathogenic bacteria have different antibiotic resistance among different types of SOT groups and specimens.Therefore,it is necessary to strengthen the pathogen examination after different types of SOT and optimize the anti infection treatment plan related to transplantation based on drug sensitivity re-sults.

Objective: To establish a mouse model of lung injury induced by self-antigens and explore its pathological mechanisms to provide a reliable animal model for studying the pathogenesis of rheumatoid arthritis-associated interstitial lung disease(RA-ILD). Methods: The mice were injected intradermal with antigenemulsion made of complete freund′s adjuvant(CFA) and lung tissue protein, and the emulsion prepared with incomplete Freund′s adjuvant(IFA) was used to enhance immunity. HE staining was used to observe the histopathological changes in the mice. Masson staining was used to detect pulmonary fibrosis. The mRNA levels of rheumatoid factor(RF), krebs von den lungen-6(KL-6) and surfactant protein D(SP-D) were detected by qPCR. The levels of ACPA, IgG1, IgG2a and IgG3 antibodies were detected by ELISA. The changes of inflammatory cells and α-smooth muscle actin(α-SMA)expression in lung tissue were detected by immunofluorescence staining. The protein expression levels of Collagen I and α-SMA were detected by Western blot. The changes of immune cells were detected by flow cytometry. Results: HE staining showed that inflammatory cell infiltration increased and tissue structure changed significantly in lung tissue after the model was established by self-lung tissue antigen. Masson staining showed increased collagen deposition in lung tissue of model mice. qPCR tests revealed elevated mRNA levels of RF, KL-6 and SP-D. ELISA tests revealed elevated levels of ACPA, IgG1 and IgG3 antibodies. Immunofluorescence results showed that monocytes and T cells increased, and α-SMA expression increased in the model group. Western blot results showed increased protein expressions of Collagen Ⅰ and α-SMA. The flow cytometry results showed an increase in T cells and monocytes in the lung tissue. Conclusion The mouse model of lung injury induced by self-antigens is successfully established, and T cells and monocytes may be involved in the occurrence and progression of the disease.

Objective: To explore the role of tumour necrosis factor-α(TNF-α) in splenic infection of mice by Leishmania major. Methods: To establish an infection model, promastigotes of Leishmania were injected intradermally into the right hind foot of mice. The thickness of the footpad and body weight were measured to monitor the infection. Histological changes in the spleen after infection were observed by HE staining. Changes in lymphocytes and monocytes in the spleen were detected by flow cytometry. The expression level of arginase-1(Arg-1) and inducible nitric oxide synthase(iNOS) in the spleen was determined by indirect immunofluorescence. The effect of TNF-α on macrophage infection with Leishmania was evaluated in vitro. Results: Compared to B6.WT mice, the spleens of B6.TNF-/- mice showed significant enlargement 42 days post-infection, with structural disruption. Various cells infiltrated and were dispersed throughout the entire spleen. Flow cytometry results indicated that after infection with Leishmania, there was no significant change in the proportions of T cells and B cells in the spleens of the mice, while CD11b monocytic cells significantly increased. Immunofluorescence results revealed that the M2 macrophage/monocyte marker Arg-1 was highly expressed in the spleens of B6.TNF-/- mice(P < 0. 05) . The expression of iNOS in the spleens of B6. WT mice was relatively strong (P < 0. 05) . In vitro studies found that the absence or inhibition of TNF⁃α significantly increased the infection of Leishmania by peritoneal macrophages ( P <0. 01), while the addition of TNF-a markedly inhibited this infection (P <0. 01). Conclusion The splenic infec-tion in B6. TNF mice following subcutaneous inoculation of Leishmania in the hind footpad may be associatedwith the absence of TNF-a, which leads to M2-type differentiation of macrophages and reduced nitric oxide producton.