The aim of this study is to investigate the immune effect of H9 subtype avian influenza virus(AIV)NP protein on mice and lay the foundation for the development of avian influenza vi-rus(AIV)vaccine.The H9N2 virus NP gene amplification product was cloned into the pET-32a expression vector,and the protein expression was verified by SDS-PAGE and Western blot,and the immune effect was evaluated by measuring the secretion of supernatant multicytokines in mouse splenocytes culture.The results showed that the total length of the coding region sequence of NP gene was 1 497 bp,NP recombinant proteins exist in both soluble and insoluble protein forms,and the specific bands were visible in Western blot.After immunizing mice,serum produces IgG-bind-ing antibodies with antibody titers of 1∶40 000.Compared with the control group,IL-2,IL-5 and IL-13 were significantly increased(P＜0.001),and the secretion of IL-6 was significantly increased compared with the control group.IL-4 and IL-12 p70 secretions were elevated compared with con-trols,but there was no significant difference.Compared with the control group,the secretions of IL-1β,IL-18,GM-CMF,TNF-α and IFN-γ were inhibited,but the difference was not significant(P＞0.05).The results showed that NP recombinant protein is a good immunogen,laying a foundation for in-depth research on influenza vaccine.

Objective:To explore changes of acid-base metabolism in the brain tissue of patients with chronic ischemic cerebrovascular disease (CICVD) using MRI amide proton transfer-weighted (APTw) imaging.Methods:This was a cross-sectional study. From January 2021 to July 2022, thirty-nine patients with CICVD at West China Hospital, Sichuan University were retrospectively included. All patients received CT perfusion (CTP) and APTw imaging. NeuBrainCARE brain perfusion software was used to analyze the impaired perfusion sites and measure the mean transit time (MTT) and time to peak (TTP). Standard spatial matching between CTP and APTw images was performed to measure the APTw values of the same sites. For comparison with normal tissue, APTw values were measured for normal-appearing white matter (NAWM) in the ipsilateral cerebral hemisphere, the contralateral cerebral hemisphere, and the ipsilateral cerebellar hemisphere in areas of impaired perfusion. ANOVA was used to compare the APTw values of impaired perfusion brain tissue, ipsilateral cerebral NAWM, contralateral cerebral NAWM, and ipsilateral cerebellar NAWM. The Bonferroni method was used to correct for multiple comparisons. Pearson correlation coefficient was used to analyze the correlation between APTw values and MTT and TTP in the cerebral tissue with impaired perfusion.Results:In 39 patients with CICVD, both the mean and minimum APTw values of cerebral tissue with impaired perfusion were significantly lower than those in the NAWM of the ipsilateral cerebral hemisphere, the contralateral cerebral hemisphere, and the ipsilateral cerebellar hemisphere ( P<0.001). In the NAWM of the cerebellar hemispheres with unimpaired perfusion, both the mean and minimum APTw values were significantly higher than those in the ipsilateral cerebral hemispheres and the contralateral cerebral hemisphere ( P<0.001). Correlation analysis showed that MTT was significantly negatively correlated with both the mean APTw and the minimum APTw ( r values were -0.90 and -0.82, P<0.001). TTP was significantly negatively correlated with both the mean APTw and the minimum APTw ( r values were -0.86 and -0.78, P<0.001). Conclusion:APTw value can reflect acidosis in cerebral tissue with impaired perfusion in patients with CICVD.

Objective:To explore the value of serum matrix metalloproteinase-1 (matrix metalloproteinase-1), and phosphatase and tensin hmmlogydeleted on ten (PTEN), in predicting osteoporosis (OP) in women with gestational diabetes mellitus (GDM) .Methods:The clinical data of.218 pregnant women with GDM treated in Linyi People’s Hospital from Sep. 2019 to Feb. 2023 were retrospectively collected and included in the GDM group. Another 98 healthy pregnant women who underwent physical examination during the same period were selected as the control group. Age, body mass index (BMI), blood glucose related indexes and serum MMP-1 and PTEN levels were compared between the two groups. According to the occurrence of osteoporosis (OP) in the second and third trimester of pregnancy, the GDM group was divided into OP group and non-OP group. Logistic regression analysis was used to explore the influencing factors of OP in pregnant women with GDM in the second and third trimesters. ROC curve was drawn to analyze the predictive value of serum MMP-1 and PTEN levels for osteoporosis in GDM pregnant women in the second and third trimesters.Results:Body mass index (BMI), fasting blood glucose (FBG), glycated hemoglobin A1c (HbA1c) and serum MMP-1 and PTEN levels in GDM group were higher than those in control group, but BUA, SOS and SI were lower than those in control group ( t=6.18, 12.68, 9.91, 11.42, 7.70, 5.63, 5.76, 9.84, all P<0.05). Among 218 GDM women, 29 cases had OP and 189 cases did not have OP. There were no significant differences in fasting blood glucose, glycosylated hemoglobin, blood calcium, blood magnesium, or blood phosphorus between the two groups (statistical value: 0.96, 0.86, 1.35, 1.06, 0.49, P>0.05). Logistic regression analysis showed that age, abnormal increase of MMP-1 and PTEN levels were independent risk factors for OP in GDM women in the second and third trimesters [ OR (95% CI) =1.260 (1.103-1.440) ,1.075 (1.031-1.120) ,1.055 (1.023-1.089), all P<0.05]. ROC curve showed that the AUC of MMP-1 and PTEN in predicting OP in GDM pregnant women in the second and third trimesters were 0.824 and 0.777, respectively. The AUC of the combination of the two in predicting OP in GDM pregnant women in the second and third trimesters was 0.933, and the specificity and sensitivity were 83.04% and 93.48%, respectively, which were higher than those of single detection. Conclusions:MMP-1 and PTEN in early pregnancy may play a potential role in the prediction of OP in pregnant women with GDM. Close monitoring of MMP-1 and PTEN levels may provide new ideas for the clinical diagnosis and treatment of OP high-risk patients.

Objective To investigate the mechanism of Yes-associated protein 1(YAP1)ameliorating aristolochic acid 1(AAI)-induced liver injury in mice based on untargeted metabolomics techniques.Methods There were 83-week-old male hepatocyte-specific Yap1 gene knockout mice(genotyped as Yap1Flox/Flox,Albumin-Cre,aka.Yap1LKO)were randomly selected as the Yap1LKO+AAI group,and 8 Yap1Flox control mice as the Yap1Flox+AAI group.Both groups were injected intraperitoneally with AAI at a dose of 2.5 mg·kg-1·d-1 for 14 consecutive days.Genotypes were identified by tail PCR;serum alanine transaminase(ALT)and aspartate transaminase(AST)activities were determined by microplate assay;histopathological changes of liver tissue were observed by HE staining;and the protein expression of YAP1 in liver tissue was determined by immunohistochemistry.The untargeted metabolomics approach was used to analyze the liver tissue differential metabolites,and the samples were analyzed by ultra performance liquid chromatography-quadrupole-electrostatic field orbit trap high-resolution mass spectrometry,and the differential metabolites were screened by principal component analysis(PCA),Partial least square-discriminant analysis(PLS-DA),and orthogonal partial least squares-discriminant analysis(OPLS-DA);using HMDB database and METLIN database to identify metabolites,and the pathway enrichment of differential metabolites was analyzed by KEGG database.Results(1)After 14 days of AAI induction,the increase of body mass in Yap1LKO mice was lower than that in Yap1Flox mice,but there was no statistical significance(P＞0.05).On day 14,compared with the Yap1Flox+AAI group,the serum ALT and AST enzyme activities in the Yap1LKO+AAI group of mice were significantly increased(P＜0.05),and the histopathological damage of the liver was significantly aggravated.The livers of the Yap1Flox mice had a positive protein expression of YAP1,whereas the Yap1LKO mice did not have a positive protein expression of YAP1.(2)A total of 139 differential metabolites with significant changes(VIP＞1 and P＜0.05)were screened by metabonomic analysis;compared with Yap1LKO+ AAI group,62 liver metabolites in Yap1Flox+AAI group were up-regulated,including choline,taurine,hypotaurine,α-linolenic acid,eleostearic acid,chenodeoxycholic acid and so on.Seventy-seven metabolites were down-regulated including glycerophosphocholine,L-phosphatidylcholine,L-glutamine,L-serine,L-glutathione,5-methionine,phenylalanine,glucose 6-phosphate,lactic acid,uric acid glycosides,etc..KEGG-enriched pathways were mainly choline metabolism,glycerophospholipid metabolism,insulin resistance,glutathione metabolism,etc..Conclusion Hepatocyte-specific Yap1 gene knockout exacerbated AAI-induced liver injury in mice,and YAP1 was involved in the regulation of choline metabolism and glycerophospholipid metabolism through the up-regulation of unsaturated fatty acids,such as choline and taurine,which ameliorated AAI-induced liver injury in mice.

ObjectiveTo investigate the material basis of homologous and heterogeneous effect of Aurantii Fructus Immaturus（AFI） and Aurantii Fructus（AF） based on the total statistical moment analysis and molecular connectivity index（MCI）. MethodRelevant literature at home and abroad and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） were consulted to establish the chemical composition database of AFI and AF， and set up their fingerprints by ultra-high performance liquid chromatography（UPLC）， and the total statistical moments and similarity parameters of the fingerprint were calculated. According to MCI， all components of AFI and AF were divided into different component groups， the average values of 0-8^th order（⁰χ-⁸χ） MCI of the common component groups of AFI and AF were calculated. ResultThe values of total zero-order moment（AUC_T） of AFI and AF were （10.57±2.45）×10⁶， （5.09±0.89）×10⁶ μV·s， the values of total first-order moment（MCRT_T） were （11.57±1.58）， （12.10±1.29） min， the values of total second-order moments（VCRT_T） were（24.49±2.30）， （26.49±2.54） min²， respectively. It showed that qualitative and quantitative parameters of AFI and AF were significantly different. The components with high similarity such as neohesperidin， hesperidin and narirutin were screened as the common potential pharmacodynamic components of AFI and AF. The non-common components of AFI， such as alysifolinone and imperatorin， and the non-common components of AF， such as neoeriocitrin and isosakuranin， with high similarity were screened out as potential heterogeneous components of AFI and AF. The composition groups of AFI and AF were classified into six categories， and the similarities between the composition groups of AFI and AF and the total constituents were 0.872-0.979 and 0.918-0.997， the average values of ⁰χ-⁸χ MCI of alkaloids in AFI and AF were 3.65 and 3.14， the average values of ⁰χ-⁸χ MCI of flavonoids were 8.47 and 8.47， the average values of ⁰χ-⁸χ MCI of volatile oils were 2.71 and 3.48， respectively. It showed that there were some differences in MCI of chemical constituents（groups） between AFI and AF. ConclusionThe chemical constituents（groups） of AFI and AF not only differ in content and species， but also in structural characteristics and structure-activity relationship， which can provide a basis for further explaining the scientific connotation of homologous and heterogeneous effect of AFI and AF.

Objective:Optimizing and upgrading the accounting methods and approaches for each dimension by comparing and analyzing the practical effects of the full-dimension costing path in public hospitals.Methods:List the mainstream accounting paths guided by the current cost accounting plan for public hospitals,to compare the advantages and disadvantages of the results of each type of accounting methods and their degree of difficulty,and to optimize and upgrade the details of accounting in all dimensions.Re-sults:Different accounting methods have their own advantages and disadvantages,and the path selection needs to rely on the scale of public hospitals,management needs,accounting level and information construction degree and other comprehensive factors,that is,with the selection of accounting paths in each dimension from simple to fine,the degree of data refinement and reference strength are correspondingly from weak to strong.Conclusion:Public hospitals should combine their own accounting basic conditions,choose the best accounting methods and paths,build a cost accounting control system,and continuously optimize and upgrade the ac-counting model,in order to provide strong decision-making support for hospitals to find out the asset base,strengthen resource allo-cation and improve data governance capabilities.

Objective:Optimizing and upgrading the accounting methods and approaches for each dimension by comparing and analyzing the practical effects of the full-dimension costing path in public hospitals.Methods:List the mainstream accounting paths guided by the current cost accounting plan for public hospitals,to compare the advantages and disadvantages of the results of each type of accounting methods and their degree of difficulty,and to optimize and upgrade the details of accounting in all dimensions.Re-sults:Different accounting methods have their own advantages and disadvantages,and the path selection needs to rely on the scale of public hospitals,management needs,accounting level and information construction degree and other comprehensive factors,that is,with the selection of accounting paths in each dimension from simple to fine,the degree of data refinement and reference strength are correspondingly from weak to strong.Conclusion:Public hospitals should combine their own accounting basic conditions,choose the best accounting methods and paths,build a cost accounting control system,and continuously optimize and upgrade the ac-counting model,in order to provide strong decision-making support for hospitals to find out the asset base,strengthen resource allo-cation and improve data governance capabilities.

Objective:Optimizing and upgrading the accounting methods and approaches for each dimension by comparing and analyzing the practical effects of the full-dimension costing path in public hospitals.Methods:List the mainstream accounting paths guided by the current cost accounting plan for public hospitals,to compare the advantages and disadvantages of the results of each type of accounting methods and their degree of difficulty,and to optimize and upgrade the details of accounting in all dimensions.Re-sults:Different accounting methods have their own advantages and disadvantages,and the path selection needs to rely on the scale of public hospitals,management needs,accounting level and information construction degree and other comprehensive factors,that is,with the selection of accounting paths in each dimension from simple to fine,the degree of data refinement and reference strength are correspondingly from weak to strong.Conclusion:Public hospitals should combine their own accounting basic conditions,choose the best accounting methods and paths,build a cost accounting control system,and continuously optimize and upgrade the ac-counting model,in order to provide strong decision-making support for hospitals to find out the asset base,strengthen resource allo-cation and improve data governance capabilities.

Objective:To understand the current status of gastroscopy in diagnosing gastric lesions in general population, and to recommend the optimal age for the first gastroscopy and intervals for repeated gastroscopy.Methods:The gastroscopy records of residents aged 18-80 years in Yinzhou District of Ningbo, Zhejiang Province, between April 2010 and December 2021 were analyzed retrospectively. The detections of gastric lesions across different years, age and genders were described. Goodness of fit tests were applied to compare the differences in detection rates of different lesions in first-time endoscopy in different age groups and different populations. Generalized additive models were used to fit the trend of age specific gastric lesion detection rate explore the optimal age for gastroscopy. The appropriate gastroscopy intervals were determined according to the progress of the gastric lesions detected in repeated gastroscopy.Results:A total of 237 751 participants with 344 398 gastroscopy records were included in analyses. A total of 5 597 cases of chronic atrophic gastritis (CAG), 9 796 cases of intestinal metaplasia (IM), 165 cases of low-grade intraepithelial neoplasia (LGIN), 52 cases of high-grade intraepithelial neoplasia (HGIN) and 435 cases of gastric cancer were detected by the first gastroscopy. The overall detection rate of gastric lesions increased significantly in age group 45-70 years, and remained stable after 70 years old, with LGIN and HGIN showing notable increases at 50 and 55 years old, respectively. Repeated gastroscopy detected CAG, IM, LGIN, and HGIN at a higher rate compared with the first gastroscopy. Normal/superficial gastritis progressed in 3-5 years, whereas CAG or more severe lesions progressed in 1-6 years.Conclusion:Gastroscopy is recommended for general population aged 45 years and above. Furthermore, gastroscopy can be performed every 3-5 years for individuals with normal endoscopy results and once a year for patients with CAG or more severe gastric lesions.

Objective:Optimizing and upgrading the accounting methods and approaches for each dimension by comparing and analyzing the practical effects of the full-dimension costing path in public hospitals.Methods:List the mainstream accounting paths guided by the current cost accounting plan for public hospitals,to compare the advantages and disadvantages of the results of each type of accounting methods and their degree of difficulty,and to optimize and upgrade the details of accounting in all dimensions.Re-sults:Different accounting methods have their own advantages and disadvantages,and the path selection needs to rely on the scale of public hospitals,management needs,accounting level and information construction degree and other comprehensive factors,that is,with the selection of accounting paths in each dimension from simple to fine,the degree of data refinement and reference strength are correspondingly from weak to strong.Conclusion:Public hospitals should combine their own accounting basic conditions,choose the best accounting methods and paths,build a cost accounting control system,and continuously optimize and upgrade the ac-counting model,in order to provide strong decision-making support for hospitals to find out the asset base,strengthen resource allo-cation and improve data governance capabilities.