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.

Clinically， the occurrence of the breaking bad news is inevitable. For patients， they not only need to accept and cope with bad news， but also need to inform the bad news to their families with clear thinking and appropriate language， seeking their support and cooperation. This paper analysed the connotation and informing dilemma of bad news， investigated patients’ informing tendencies， as well as evaluated the advantages and disadvantages of three disclosure methods， including concealment， immediate informing， and staged informing. On these bases， a detailed response strategy for patients to inform their families was proposed in three parts， including pre-preparation， mid-articulation， and post-summary. In the initial phase， thorough preparation is essential. During the middle stage， when delivering bad news， use plain language and help family members adjust their emotions. In the final phase， ensure that all information and viewpoints have been fully communicated. In addition， the roles and analysis steps that doctors should play were analysed from their perspective and combined with the degree of doctor-patient trust. Effective informing of bad news is not only about communication skills， but also involves a deep understanding and respect for the psychological needs of patients and their families. Through meticulous preparation， appropriate expression， emotional support， and clear confirmation， communication and trust are promoted to face and overcome difficulties together.

To provide in-service medical technicians and nurses with convenient access to continuing education resources in transfusion medicine, reduce transfusion-related adverse events, and ensure the safety, rationalization, and effectiveness of clinical transfusion, we designed and developed an online transfusion continuing education platform. The platform was based on the new managed code programming model.NET Core and the powerful functions of hypertext preprocessor PHP 7.4, addressing current issues in transfusion online continuing education. Through in-depth analysis of student attributes, learning behaviors, and teaching behaviors, a comprehensive online continuous teaching quality evaluation index system was established. This system not only facilitates the quantitative assessment of teaching quality but also successfully integrates the two core functions of teaching and management, thereby achieving unified online teaching.

Objective:To analyze risk factors for early trauma-induced coagulopathy (TIC) in pediatric patients with moderate-to-severe traumatic brain injury (msTBI), develop a predictive model and evaluate its predictive performance.Methods:A retrospective cohort study was conducted to analyze the clinical data of 290 pediatric patients with msTBI who were admitted to Children′s Hospital of Soochow University between January 2016 and December 2024, including 188 boys and 102 girls, aged 0.2-15.7 years [5.2(2.8, 9.3)years]. Based on the coagulation test results at admission, the patients were divided into TIC group ( n=162) and non-TIC group ( n=128). The patients were randomly allocated into training set ( n=203) and validation set ( n=87) at a ratio of 7∶3. Demographic characteristics, clinical data, vital signs, imaging findings, arterial blood gas analysis results, and coagulation profiles of the patients were collected. Univariate analysis and Lasso regression analysis were used to identify risk factors associated with early TIC in children with msTBI and multivariate Logistic regression analysis was performed to determine independent risk factors and construct a predictive model. The model′s discrimination and calibration were evaluated using the area under the receiver operating characteristic (ROC) curve (AUC), Hosmer-Lemeshow (H-L) test, and calibration curve. Its clinical utility was assessed through decision curve analysis (DCA). Results:Significant differences were observed between the TIC group and non-TIC group in terms of age, weight, time from injury to admission, child′s Glasgow coma scale (CGCS) score, pediatric trauma score (PTS), shock index, heart rate, respiratory rate, systolic blood pressure, Rotterdam CT score, intraventricular hemorrhage, cerebral contusion, brain herniation, long bone fracture, pelvic fracture, hemopneumothorax, pulmonary contusion, intra-abdominal organ injury, actual bicarbonate, base excess, base excess in the extracellular fluid, blood glucose, hemoglobin (Hb), osmolarity, blood calcium, anion gap, blood lactate, prothrombin time, activated partial thromboplastin time, international normalized ratio, and platelet count ( P<0.05). With coagulation-related variables excluded, the following features were identified with Lasso regression including CGCS score, PTS, heart rate, systolic blood pressure, long bone fracture, blood glucose, and Hb. Multivariate Logistic regression analysis revealed that CGCS score≤8 points ( OR=3.05, 95% CI 1.65, 5.63), PTS>5 points ( OR=0.45, 95% CI 0.23, 0.89), systolic blood pressure ( OR=0.98, 95% CI 0.97, 0.99), blood glucose ( OR=1.09, 95% CI 1.01, 1.17), and long bone fracture ( OR=2.47, 95% CI 1.13, 5.42) were influencing factors for early TIC in children with msTBI ( P<0.05). The regression equation of the predictive model was established as follows: Logit[ P/(1- P)]=1.01×"CGCS score≤8 points"-0.69×"PTS>5 points"- 0.02×"systolic blood pressure"+0.89×"long bone fracture"+0.08×"blood glucose"+1.32. The ROC curve analysis showed that the training set had an AUC of 0.86 (95% CI 0.78, 0.94), with sensitivity and specificity of 76.6% and 92.5%, while the AUC was 0.80 (95% CI 0.74, 0.86), with sensitivity and specificity of 75.7% and 79.6% in the validation set. H-L test results showed a χ2 value of 8.18 ( P=0.416) in the training set and 5.30 ( P=0.216) in the validation set. The calibration curves for both sets demonstrated good agreement with the actual curves, indicating that the predicted probabilities closely matched the observed probabilities. The DCA results indicated that both the training set and validation set demonstrated positive net benefits within threshold probabilities ranges of 10%-100% and 15%-96%. Conclusions:Independent risk factors for early TIC in pediatric msTBI patients include CGCS score≤ 8 points, PTS≤5 points, low systolic blood pressure, long bone fracture, and high blood glucose. The predictive model constructed based on these factors demonstrates favorable predictive performance and clinical application value.

Aim To explore the mechanism of the syn-ergistic effect of the ferroptosis inducer erastin com-bined with shikonin in promoting ferroptosis in human hypertrophic scar fibroblasts(HSFBs).Methods Hypertrophic scar tissues provided by the General Hos-pital of Ningxia Medical University were collected,and HSFBs were extracted.HSFBs were identified by HE staining and immunofluorescence.The inhibitory rates of Era and SHK on HSFBs at different concentrations were detected by CCK-8 assay,and the IC50 value was calculated.CompuSyn software was used to calculate the co-use index(CI).Control group,Erastin(Era)group,shikonin(SHK)group and Era+SHK group were set up,and the number and morphological chan-ges of cells were observed after 24 hours of interven-tion.The ability of cell migration and invasion was de-tected by scratch test and Transwell test.The changes of malondialdehyde(MDA),total iron ion and reactive oxygen species(ROS)were detected by corresponding biochemical kits.The expressions of collagen I,α-SMA and GOT1,SLC7A11,GPX4 and FTH1 were detected by Western blot.Results The IC50 value of Era and SHK of primary HSFBs was 2.22 μmol·L-1 and 3.94μmol·L-1 respectively,which was used as the single drug concentration for subsequent experiments.The CompuSyn software was employed to calculate the CI value when the two drugs were used in combination,and the concentrations corresponding to CI=0.39597(Era:1.2 μmol·L-1+SHK:1.5 μmol·L-1)were selected as subsequent combination concentrations(Because when CI was equal to 0.395 97,the concen-tration of each drug was lower than the concentration of single drug,and the inhibition rate of combined drug was greater than 50％).Compared with the monother-apy group,the number of HSFBs in the SHK+Era group was significantly reduced,cell membrane showed breakage and vesiculation,cell wrinkling became smal-ler,and cytoplasm was concentrated.The migration and invasion ability of HSFBs in the SHK+Era group were obviously weakened(P＜0.05),and the expres-sion of fibrosis-related proteins collagen Ⅰ and α-SMA was reduced(P＜0.05);the contents of MDA,total i-ron ions,and ROS in HSFBs of the SHK+Era group increased(P＜0.05),and the protein expression lev-els of SLC7A11,GOT1,GPX4,and FTH1 further de-creased(P＜0.05).Conclusions Erastin in combi-nation with shikonin can synergistically inhibit the pro-liferation,migration and fibrosis levels of HSFBs.The mechanism may be that erastin enhances the inhibition of shikotin on GOT1,increases the levels of cellular i-ron ions,ROS,and lipid peroxides,thereby promoting ferroptosis in HSFBs.

Objective:To analyze risk factors for early trauma-induced coagulopathy (TIC) in pediatric patients with moderate-to-severe traumatic brain injury (msTBI), develop a predictive model and evaluate its predictive performance.Methods:A retrospective cohort study was conducted to analyze the clinical data of 290 pediatric patients with msTBI who were admitted to Children′s Hospital of Soochow University between January 2016 and December 2024, including 188 boys and 102 girls, aged 0.2-15.7 years [5.2(2.8, 9.3)years]. Based on the coagulation test results at admission, the patients were divided into TIC group ( n=162) and non-TIC group ( n=128). The patients were randomly allocated into training set ( n=203) and validation set ( n=87) at a ratio of 7∶3. Demographic characteristics, clinical data, vital signs, imaging findings, arterial blood gas analysis results, and coagulation profiles of the patients were collected. Univariate analysis and Lasso regression analysis were used to identify risk factors associated with early TIC in children with msTBI and multivariate Logistic regression analysis was performed to determine independent risk factors and construct a predictive model. The model′s discrimination and calibration were evaluated using the area under the receiver operating characteristic (ROC) curve (AUC), Hosmer-Lemeshow (H-L) test, and calibration curve. Its clinical utility was assessed through decision curve analysis (DCA). Results:Significant differences were observed between the TIC group and non-TIC group in terms of age, weight, time from injury to admission, child′s Glasgow coma scale (CGCS) score, pediatric trauma score (PTS), shock index, heart rate, respiratory rate, systolic blood pressure, Rotterdam CT score, intraventricular hemorrhage, cerebral contusion, brain herniation, long bone fracture, pelvic fracture, hemopneumothorax, pulmonary contusion, intra-abdominal organ injury, actual bicarbonate, base excess, base excess in the extracellular fluid, blood glucose, hemoglobin (Hb), osmolarity, blood calcium, anion gap, blood lactate, prothrombin time, activated partial thromboplastin time, international normalized ratio, and platelet count ( P<0.05). With coagulation-related variables excluded, the following features were identified with Lasso regression including CGCS score, PTS, heart rate, systolic blood pressure, long bone fracture, blood glucose, and Hb. Multivariate Logistic regression analysis revealed that CGCS score≤8 points ( OR=3.05, 95% CI 1.65, 5.63), PTS>5 points ( OR=0.45, 95% CI 0.23, 0.89), systolic blood pressure ( OR=0.98, 95% CI 0.97, 0.99), blood glucose ( OR=1.09, 95% CI 1.01, 1.17), and long bone fracture ( OR=2.47, 95% CI 1.13, 5.42) were influencing factors for early TIC in children with msTBI ( P<0.05). The regression equation of the predictive model was established as follows: Logit[ P/(1- P)]=1.01×"CGCS score≤8 points"-0.69×"PTS>5 points"- 0.02×"systolic blood pressure"+0.89×"long bone fracture"+0.08×"blood glucose"+1.32. The ROC curve analysis showed that the training set had an AUC of 0.86 (95% CI 0.78, 0.94), with sensitivity and specificity of 76.6% and 92.5%, while the AUC was 0.80 (95% CI 0.74, 0.86), with sensitivity and specificity of 75.7% and 79.6% in the validation set. H-L test results showed a χ2 value of 8.18 ( P=0.416) in the training set and 5.30 ( P=0.216) in the validation set. The calibration curves for both sets demonstrated good agreement with the actual curves, indicating that the predicted probabilities closely matched the observed probabilities. The DCA results indicated that both the training set and validation set demonstrated positive net benefits within threshold probabilities ranges of 10%-100% and 15%-96%. Conclusions:Independent risk factors for early TIC in pediatric msTBI patients include CGCS score≤ 8 points, PTS≤5 points, low systolic blood pressure, long bone fracture, and high blood glucose. The predictive model constructed based on these factors demonstrates favorable predictive performance and clinical application value.

Objective To investigate the nebulization characteristics of human interferon α1b for injection.To characterize and compare the delivery rate,total drug substance and the aerodynamic characteristics between the two types of nebulizer.Methods Connect two types of nebulizers with a breathing simulator,respectively,and simulate the breathing patterns of an infant and child.Measure the delivery rate,total delivered dose,and delivery uniformity.The aerodynamic properties of human interferon α1b for injection were evaluated by the next generation impactor(NGI).The content of human interferon α1b was quantified by double antibody sandwich ELISA.Results In the three batches of samples in infant mode,the delivery rate and total delivered dose determind by A nebulizer were 0.45,0.49,0.44 μg·min-1,3.06,3.21,3.81 μg,respectcively;and 0.12,0.14,0.16 μg·min-1,0.73,0.73,0.75 μg,respectcively by B nebulizer.In child mode,the delivery rate and total delivered doses determined by A nebulizer were 1.36,1.49,1.20 μg·min-1,7.44,7.17,and 6.54 μg,respectcively;and 0.37,0.36,0.43 μg·min-1,1.66,1.59,and 1.41 μg,respectcively by B nebulizer.In child and infant mode,the ten results of the total drug substance delivered determined by nebulizer A were both between 65%to 135%of the average.The FPD,FPF,MMAD,and GSD determined by A neublizer of three batch samples were 2.48,2.92,2.35 μg,59.0%,57.4%,59.1%,4.18,4.34,4.15 μm,1.94,1.98,2.01,respectively.The FPD,FPF,MMAD and GSD determined by B neublizer of three batches samples were 2.70,3.38,3.06 μg,67.6%,66.4%,66.3%,3.55,3.65,3.68 μm,2.03,2.04,2.06,respectively.Conclusions The data obtained in this research characterized the in vitro nebulization characteristics of human interferon α1b for injection and provided a theoretical basis and reference for in vitro study and clinical practice.The influence of different types of nebulizers on nebulization characteristics was evaluated as well.It is suggested that the quality standard of nebulizers be strictly formulated and the use of nebulizers be standardized.

Aim To investigate the effect of p-AMPK activity on autophagy in different subtypes of MDA-MB-231(triple-negative breast cancer cells)and MCF-7(estrogen receptor-positive cells)and its regulatory mechanism.Methods MDA-MB-231 cells were trea-ted with EBSS,Baf-A1,and EBSS+Baf-A1 for four hours,and MCF-7 cells for eight hours.The effects of autophagy on cell proliferation and apoptosis were ob-served,mitochondrial morphology was examined,and the expression of autophagy markers LC3B,P62,LAMP1,TOM20,AMPK,p-AMPK,ULK1,and Bec-lin1/VPS34 proteins was detected.The autophagy pathway was validated by inhibiting AMPK activity.Results Breast cancer cells underwent autophagy af-ter starvation induction(EBSS),with inconsistent au-tophagy processes observed in different subtypes of breast cancer cells.Autophagy inhibited cell prolifera-tion.In MDA-MB-231 cells,autophagy led to an in-crease in p-AMPK levels and a decrease in ULK1 lev-els,initiating autophagy through p-AMPK activation of ULK1.In MCF-7 cells,both p-AMPK and ULK1 levels decreased after autophagy,suggesting that autophagy might not be mediated by p-AMPK activation.Conclu-sions MDA-MB-231 cells primarily initiate autophagy by directly activating ULK1 by p-AMPK,independent of the MTOR pathway.In MCF-7 cells autophagy might be triggered by inhibiting MTOR through AMPK activity or directly activating MTOR through other up-stream factors.Regulating p-AMPK activity based on the autophagy pathways in different cell subtypes could enable more precise targeting and treatment of different types of breast cancer.

Objective To investigate the nebulization characteristics of human interferon α1b for injection.To characterize and compare the delivery rate,total drug substance and the aerodynamic characteristics between the two types of nebulizer.Methods Connect two types of nebulizers with a breathing simulator,respectively,and simulate the breathing patterns of an infant and child.Measure the delivery rate,total delivered dose,and delivery uniformity.The aerodynamic properties of human interferon α1b for injection were evaluated by the next generation impactor(NGI).The content of human interferon α1b was quantified by double antibody sandwich ELISA.Results In the three batches of samples in infant mode,the delivery rate and total delivered dose determind by A nebulizer were 0.45,0.49,0.44 μg·min-1,3.06,3.21,3.81 μg,respectcively;and 0.12,0.14,0.16 μg·min-1,0.73,0.73,0.75 μg,respectcively by B nebulizer.In child mode,the delivery rate and total delivered doses determined by A nebulizer were 1.36,1.49,1.20 μg·min-1,7.44,7.17,and 6.54 μg,respectcively;and 0.37,0.36,0.43 μg·min-1,1.66,1.59,and 1.41 μg,respectcively by B nebulizer.In child and infant mode,the ten results of the total drug substance delivered determined by nebulizer A were both between 65%to 135%of the average.The FPD,FPF,MMAD,and GSD determined by A neublizer of three batch samples were 2.48,2.92,2.35 μg,59.0%,57.4%,59.1%,4.18,4.34,4.15 μm,1.94,1.98,2.01,respectively.The FPD,FPF,MMAD and GSD determined by B neublizer of three batches samples were 2.70,3.38,3.06 μg,67.6%,66.4%,66.3%,3.55,3.65,3.68 μm,2.03,2.04,2.06,respectively.Conclusions The data obtained in this research characterized the in vitro nebulization characteristics of human interferon α1b for injection and provided a theoretical basis and reference for in vitro study and clinical practice.The influence of different types of nebulizers on nebulization characteristics was evaluated as well.It is suggested that the quality standard of nebulizers be strictly formulated and the use of nebulizers be standardized.

Aim To investigate the effect of p-AMPK activity on autophagy in different subtypes of MDA-MB-231(triple-negative breast cancer cells)and MCF-7(estrogen receptor-positive cells)and its regulatory mechanism.Methods MDA-MB-231 cells were trea-ted with EBSS,Baf-A1,and EBSS+Baf-A1 for four hours,and MCF-7 cells for eight hours.The effects of autophagy on cell proliferation and apoptosis were ob-served,mitochondrial morphology was examined,and the expression of autophagy markers LC3B,P62,LAMP1,TOM20,AMPK,p-AMPK,ULK1,and Bec-lin1/VPS34 proteins was detected.The autophagy pathway was validated by inhibiting AMPK activity.Results Breast cancer cells underwent autophagy af-ter starvation induction(EBSS),with inconsistent au-tophagy processes observed in different subtypes of breast cancer cells.Autophagy inhibited cell prolifera-tion.In MDA-MB-231 cells,autophagy led to an in-crease in p-AMPK levels and a decrease in ULK1 lev-els,initiating autophagy through p-AMPK activation of ULK1.In MCF-7 cells,both p-AMPK and ULK1 levels decreased after autophagy,suggesting that autophagy might not be mediated by p-AMPK activation.Conclu-sions MDA-MB-231 cells primarily initiate autophagy by directly activating ULK1 by p-AMPK,independent of the MTOR pathway.In MCF-7 cells autophagy might be triggered by inhibiting MTOR through AMPK activity or directly activating MTOR through other up-stream factors.Regulating p-AMPK activity based on the autophagy pathways in different cell subtypes could enable more precise targeting and treatment of different types of breast cancer.