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.

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 explore the effect and mechanism of annexin A1 mimic peptide Ac2-26 on ferroptosis in hu-man umbilical vein endothelial cells(HUVEC).Methods Induction of HUVEC ferroptosis was achieved by the clas-sical ferroptosis agonist RSL3,with subsequent intervention by the annexin A1 mimtic peptide Ac2-26.The cell number and viability were detected by CCK-8 kit,the levels of malondialdehyde(MDA)and glutathione(GSH)were detected by ELISA,the expression of ferroptosis-related molecules and adhesion molecules was detected by Western blot,the lipid re-active oxygen species(ROS)levels were detected by C11-BODIPY fluorescent probe,and the mitochondrial reactive oxy-gen species(mtROS)levels were detected by MitoSOX probe.FeRhoNOX-1 fluorescent probe was used to detect intra-cellular Fe2+content,perspective microscopy was used to observe mitochondrial morphology,JC-1 fluorescent probe was used to detect mitochondrial membrane potential,kit was used to detect ATP levels,the Scratch assay was used to detect cell migration ability,and nitrate reductase assay was used to detect nitric oxide(NO)level.Results Ac2-26 inhibi-ted RSL3-induced decrease in HUVEC viability,up-regulated the expression of suppressor of ferroptosis proteolytic carrier family 7 member 11(SLC7A11),GPX4,and ferritin heavy chain 1(FTH1),increased the GSH content,decreased the MDA content,reduced the generation of intracellular lipid ROS,and decreased the intracellular Fe2+aggregation(P＜0.05 or P＜0.01);Ac2-26 inhibited RSL3-induced damage to HUVEC mitochondrial morphology and function,up-regulated ATP content(P＜0.05)and mitochondrial membrane potential(P＜0.001);Ac2-26 inhibited RSL3-induced decrease in HUVEC migratory ability,up-regulated NO levels,inhibited intercellular adhesion molecule-1(ICAM-1)and interleukin-1β(IL-1β)protein expression(P＜0.05 or P＜0.01).Conclusion Ac2-26 inhibits RSL3-induced ferroptosis in HUVEC and maintains mitochondrial morphology and function,as well as HUVEC function.

Objective This study aims to explore the efficacy of tetrahydrocurcumin(THC),the major active metabolite of curcumin,on high glucose(HG)-induced human platelet aggregation and activation as well as to clarify the underlying mechanisms in vitro.Methods Purified platelets prepared from healthy subjects were pre-incubated with various concentrations of THC(0.5 μmol/L,1 μmol/L or 10 μmol/L)or vehicle control(0.05％DMSO)for 40 min at 37℃,followed by the stimulation of normal glucose(NG,5 mmol/L)or HG(25 mmol/L)for additional 90 min.The maximal aggregation rate was determined by an aggregometer.Flow cytometry was used to measure platelet surface expression of CD62P(a typical marker of platelet activation)and generation of total intraplatelet reactive oxygen species(ROS).Meanwhile,the phosphorylation level of platelet p53 was detected by Western blot assay.Results Compared with NG group,HG intervention significantly increased platelet aggrega-tion(P<0.05)and CD62P expression(P<0.001),which were greatly inhibited by different concentrations of THC(P<0.05).Mechanistically,when compared with solvent control,THC significantly decreased the level of total ROS production(P<0.001)and p53 phosphorylation(P<0.05).In addition,HG-induced total intraplatelet ROS generation(P<0.001)and p53 phosphorylation(P<0.05)were greatly attenuated by adding a ROS scavenger N-acetyl-L-cysteine(NAC).The combination of NAC with THC(10 μmol/L)showed no additive inhibitory effects(P>0.05).Moreover,platelet aggregation and activation induced by HG were greatly decreased by NAC and a p53 specific inhibitor PFT-μ(P<0.05).The combination of THC(10 μmol/L)and NAC resulted no additive inhibitory effects on HG-increased platelet aggregation and activation(P>0.05).THC(10 μmol/L)exhibited additive inhibitory effects on platelet aggregation(P<0.05)but no additive inhibitory effects on platelet activation when combined with PFT-μ(P>0.05).Conclusions THC exerts a protective effect on HG-induced platelet aggregation and activation possibly through down-regulating ROS/p53 signaling pathway in human platelets in vitro.The current study may provide potential value for THC to improve thrombosis in diabetes mellitus and the related chronic metabolic diseases.

ObjectiveTo develop a scale for evaluating the syndrome of internal retention of dampness and turbidity in nonalcoholic fatty liver disease （NAFLD） that combines disease and syndrome and has the characteristics of traditional Chinese medicine （TCM）. MethodsAn item pool was established for evaluating the syndrome of internal retention of dampness and turbidity in nonalcoholic fatty liver disease （NAFLD） with reference to the guideline for developing international scales. A clinical survey was conducted among the outpatients and inpatients who were diagnosed with NAFLD and had the syndrome of internal retention of dampness and turbidity in Department of Hepatology and Spleen-Stomach， The First Affiliated Hospital of Henan University of Chinese Medicine， from June to August， 2023， and the items were screened based on the classical test theory and the item response theory. An expert questionnaire was developed， and expert discussions were conducted using the Delphi method to identify the items for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD. Finally， the scale was given scientific scores. ResultsA preliminary item pool was established， with 16 primary items and 22 secondary items， and it was divided into the two dimensions of disease and syndrome type. Clinical pre-survey suggested to retain 9 primary items and 14 secondary items， while the Delphi expert questionnaire recommended to retain 11 primary items and 15 secondary items， and tongue manifestation and pulse manifestation were no longer used for assessing the severity of syndrome. After hierarchical analysis and scientific assignment of scores， the scale for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD had a total score of 123 points and 9 important items， i.e.， discomfort in the hypochondrium， abdominal fullness and distension， obesity， heaviness of the head and body， loose stool， anorexia， coughing up phlegm， nausea with a tendency to vomit， and lethargy. ConclusionA preliminary scale is established for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD， which fills the gap in this research field and provides a basis for further clinical application.

[Purpose]To analyze the disease burden of malignant tumors and its changing trends in Chinese and global non-smoking female population from 1990 to 2021.[Methods]Data of mortality and disability-adjusted life year(DALY)due to malignant tumors for Chinese and global non-smoking female malignant tumors from 1990 to 2021 were extracted from the Global Burden of Disease Study 2021(GBD 2021),and the average annual percentage change(AAPC)were calculated using Joinpoint regression model.[Results]From 1990 to 2021,the number of deaths for malignant tu-mors in Chinese non-smoking female population increased from 13.7 1×104 to 26.8 1×104,with a higher increased trend compared with the global(China:AAPC=2.19％,95％CI:2.06％～2.33％;Global:AAPC=1.92％,95％CI:1.80％～2.04％,P=0.003);the age-standardized mortality rate decreased from 32.42/105 to 24.58/105,with a higher decreased trend compared with the global(China:AAPC=-0.88％,95％CI:-1.00％～-0.76％;Global:AAPC=-0.59％,95％CI:-0.68％～-0.51％,P＜0.001).From 1990 to 2021,the DALY for malignant tumors in Chinese non-smoking female population increased from 412.96×104 to 691.20×104 person-years,with a similar changing trend compared with the global(China:AAPC=1.68％,95％CI:1.56％～1.81％,Global:AAPC=1.63％,95％CI:1.52％～1.75％,P=0.536);the age-standardized DALY rate in Chinese non-smoking female population decreased from 889.58/105 to 642.65/105,with a higher decreased trend compared with the global(China:AAPC=-1.04％,95％CI:-1.15％～-0.92％;Global:AAPC=-0.69％,95％CI:-0.78％～-0.61％,P＜0.001).The top five malignant tumors of high age-standardized mor-tality rate in Chinese non-smoking female population in 2021 were tracheal,bronchus and lung cancer,colon and rectum cancer,cervical cancer,breast cancer,and liver cancer.The top five malignant tumors of high age-standardized mortality rate globally in 2021 were cervical cancer,colon and rectum cancer,breast cancer,tracheal,bronchus and lung cancer,and pancreatic cancer.The age-standardized mortality rate and DALY rate of breast cancer,liver cancer,pan-creatic cancer and corpus cancer showed overall upward trends(all P＜0.05).[Conclusion]From 1990 to 2021,the number of deaths and DALY of malignant tumors in Chinese and global non-smoking female population showed overall increased trends,and age-standardized mortality rate and DALY rate showed overall decreased trends.In future,more targeted cancer prevention measures are needed to reduce the disease burden of malignant tumors in non-smoking female population.

Objective To investigate the effects of galangin on the migration and invasion abilities of cervical cancer Hela cells and its potential mechanisms.Methods Hela cells were treated with different concentrations of galangin(0,5,10,20,40,60,80,100 μmol/L)for 48 hours,and CCK-8 assay was used to assess the impact of galangin on cell viability and to determine the half-maximal lethal concentration(IC50)of galangin.Hela cells were divided into a control group(0 μmol/L)and a galangin group(40 μmol/L treatment).Scratch wound healing assays and Transwell chamber assays were conducted to evaluate the migration and invasion abilities of the cells in each group.Western Blot was used to detect the protein expression of E-cadherin and N-cadherin.DIA quantitative proteomics technology was used to detect and screen the differentially expressed proteins between the two groups.Biological function enrichment analysis of the differential genes was performed using the KEGG Pathway and Gene Set Enrichment Analysis(GSEA)methods.Western Blot was used to verify the expression levels of Hippo/YAP signaling pathway-related proteins YAP and p-YAP.Results Compared to the control group,galangin(40 μmol/L)significantly inhibited the viability of Hela cells in a concentration-dependent manner(P<0.001).Compared with the control group,the scratch healing ability and invasion ability of cervical cancer Hela cells treated with galangin(40 μmol/L)were significantly reduced(P<0.001).The expression of E-cadherin protein was increased(P<0.05)and the expression of N-cadherin protein was decreased(P<0.001)in the galangin group(40 μmol/L)compared to the control group.KEGG and GSEA enrichment results indicated that the inhibition of malignant progression in cervical cancer by galangin was significantly associated with the Hippo/YAP signaling pathway.Western Blot confirmed that the expression level of the hallmark protein p-YAP in the Hippo signaling pathway was increased(P<0.01),while the expression level of YAP protein was decreased(P<0.05).Conclusion Galangin inhibits the proliferation,migration and invasion abilities of Hela cells in a dose-dependent manner.The underlying mechanism might be associated with the activation of the Hippo/YAP signaling pathway.

Objective To investigate the differential expression genes(DEGs)related to angiogenesis in rosacea(RA)by utilizing bioinformatics analysis in order to screen the key genes and verify their mRNA expression levels.Methods The gene microarray dataset GSE65914 was retrieved from the Gene Expression Omnibus(GEO)repository.Analyzed by R programming,the dataset was refined to identify DEGs related to RA,and then cross-referenced with angiogenesis-related genes from the GeneCards database to get a subset specific to RA angiogenesis.The process of identifying key genes was augmented by employing protein-protein interaction(PPI)network analysis and Cytoscape-based computational algorithms.The mRNA expression levels of the aforementioned pivotal genes were detected by real-time fluorescent quantitative reverse transcription PCR(RT-qPCR).Results A total of 947 RA-associated DEGs were identified from GEO dataset,and then 202 genes related to RA angiogenesis were further delineated.PPI network analysis and Cytoscape algorithm finally identified 3 key genes,that is,CXCL8,IL-1B,and STAT1.The results of RT-qPCR showed that the mRNA expression levels of MIP-2,GCP-2,IL-1B and STAT1 in RA lesions were significantly higher than those in normal controls(P<0.05).Conclusion With aid of bioinformatics analysis,our study has screened and validated key genes associated with angiogenesis in RA,namely CXCL8,IL-1B,and STAT1,which providing a theoretical basis for elucidating the potential mechanisms underlying RA-induced angiogenesis and developing targeted therapeutic strategies.

Checkpoint blockade immunotherapy has emerged as a transformative approach in cancer treatment by activating tumor-infiltrating T cells. However, the efficacy of PD-L1 blockade is restricted in "cold" tumors, which are characterized by low immunogenicity, presenting a challenge to immunotherapy. This study introduces an innovative strategy, utilizing cathepsin-cleavable N-(2-hydroxypropyl) methacrylamide (HPMA) polymer-assisted combined photodynamic therapy (PDT) and PD-L1 degradation for the first time, effectively treating T cell-deficient tumors. The degradable main-chain polymer, conjugated with photosensitizer porphyrin, facilitates the accumulation of reactive oxygen species (ROS), triggering immunogenic cell death (ICD) and promoting cytotoxic T lymphocytes (CTLs) infiltration into tumors. Multivalent peptide antagonists of PD-L1 promote PD-L1 degradation in lysosomes through receptor crosslinking, overcoming the adaptive cycling of PD-L1 to the tumor cell surface. These findings demonstrate that polymer-assisted PDT and PD-L1 crosslinking degradation represent a potential novel strategy for anti-tumor immunotherapy, providing valuable tools for expanding immunotherapy applications in immunosuppressive cancers.

Objective To investigate correlations of dynamic changes in serum C-reactive pro-tein(CRP),procalcitonin(PCT),and neutrophil-to-lymphocyte ratio(NLR)before and after symptomatic treatment with prognosis of neonates with neonatal respiratory distress syndrome(NRDS)of different etiologies.Methods A total of 110 premature infants were selected as study subjects and divided into neonatal infection group(group A)and fetal intrauterine distress group(group B)based on different causes of NRDS.Additionally,30 neonates with NRDS caused solely by prematurity were selected as control group.Serum CRP and PCT levels and NLR were compared among the three groups before and after treatment.Results There were statistically significant differences in oxygen therapy duration,ventilation duration,the proportion of infants requiring re-intubation,hospital stay,and the number of apnea episodes among three groups(P＜0.006).Before treatment,there was no sta-tistically significant difference in serum CRP levels between group A and group B(P＞0.05).Ser-um CRP levels in group B were higher than those in the control group.Moreover,serum PCT levels was lower than the group A,and NLR in the group B were higher than those in the group A and the control group.CRP,PCT levels,and NLR in the group A were higher than those in the control group(P＜0.05 or P＜0.006).After treatment,serum CRP levels in all three groups decreased compared to before treatment.Serum PCT levels in the group A decreased compared to before treat-ment.NLR in both group A and group B decreased compared to before treatment,with statistically significant differences(P＜0.006).There were statistically significant differences in serum CRP and PCT levels between the group A and the control group after treatment(P＜0.006).After treat-ment,there was a statistically significant difference in serum CRP levels between the group B and the control group(P＜0.006),but no statistically significant differences in PCT levels and NLR(P＞0.05).Compared with group A,there was a statistically significant difference in PCT levels in the group B(P＜0.006).Multivariate Logistic regression results showed that CRP,PCT,and NLR were independent influencing factors for the prognosis of infants in the group A.The receiver operat-ing characteristic curve analysis results showed that the areas under the curve(AUC)after treatment for serum CRP,PCT,and NLR alone in predicting prognosis in the group A were 0.789,0.738,and 0.758,respectively,and the AUC for combined prediction was 0.934.In the group B,the AUC for serum CRP,PCT,and NLR alone in predicting prognosis after treatment were 0.719,0.772,and 0.768,respectively,and the AUC for combined prediction was 0.886.The sensitivity and specificity of combined prediction in both groups were higher than those of each indicator alone,and the predictive value of their combined detection for the prognosis of premature infants in group A was higher than that in the group B.Conclusion There are differences in PCT levels and NLR before treatment among neonates with NRDS of different etiologies.After symptomatic treatment,CRP,PCT levels and NLR decrease in all three groups,indicating a good prognosis.