ObjectiveTo explore the changes in color， odor and chemical components during wine-processing of Chuanxiong Rhizoma（CR）， identify differential markers， and provide a basis for standardizing the process and establishing quality standards. MethodsFifteen batches of CR samples from 4 producing areas were collected. Colorimeter and electronic nose were used to detect the color changes and odor components of CR before and after wine-processing. Multivariate statistical methods including partial least squares-discriminant analysis（PLS-DA）， principal component analysis（PCA）， discriminant factor analysis（DFA） and Fisher discriminant analysis were applied to identify wine-processed CR at different processing stages and establish discriminant models， and differential components were screened out based on variable importance in the projection（VIP） value1. Then， high performance liquid chromatography（HPLC） was employed to detect the content changes of four components（ferulic acid， senkyunolide I， senkyunolide A and ligustilide） during the processing stages. ResultsThe differences of wine-processed CR at various stages were primarily reflected in color parameters L^*（brightness value）， a^*（red-green value） and b^*（yellow-blue value）. Based on chromaticity differences， the color reference ranges were established for moderately processed CR， including L^* of 46.75-48.24， a^* of 5.37-6.07 and b^* of 20.32-21.70. In odor analysis， DFA revealed significant differences among processing stages， and 11 odor markers were identified， with four differential markers（4-hydroxy-3-butylphthalide， isopropyl butyrate， L-limonene and 1-methoxyhexane） based on VIP values. HPLC results showed that there was no significant difference of the four components except for ligustilide in wine-processed CR at different stages. ConclusionThis study achieved rapid identification of wine-processed CR with different processing degrees by electronic sensory technology and differential component content detection， with discrimination accuracy rates of 92.4% and 93.272% for color and odor， respectively. This paper also established the reference ranges of main colorimetric parameters for wine-processed CR at different stages， and four differential components were screened out， providing a basis for standardizing the processing of wine-processed CR and establishing quality standards for this decoction pieces.

ObjectiveTo explore the changes in color， odor and chemical components during wine-processing of Chuanxiong Rhizoma（CR）， identify differential markers， and provide a basis for standardizing the process and establishing quality standards. MethodsFifteen batches of CR samples from 4 producing areas were collected. Colorimeter and electronic nose were used to detect the color changes and odor components of CR before and after wine-processing. Multivariate statistical methods including partial least squares-discriminant analysis（PLS-DA）， principal component analysis（PCA）， discriminant factor analysis（DFA） and Fisher discriminant analysis were applied to identify wine-processed CR at different processing stages and establish discriminant models， and differential components were screened out based on variable importance in the projection（VIP） value1. Then， high performance liquid chromatography（HPLC） was employed to detect the content changes of four components（ferulic acid， senkyunolide I， senkyunolide A and ligustilide） during the processing stages. ResultsThe differences of wine-processed CR at various stages were primarily reflected in color parameters L^*（brightness value）， a^*（red-green value） and b^*（yellow-blue value）. Based on chromaticity differences， the color reference ranges were established for moderately processed CR， including L^* of 46.75-48.24， a^* of 5.37-6.07 and b^* of 20.32-21.70. In odor analysis， DFA revealed significant differences among processing stages， and 11 odor markers were identified， with four differential markers（4-hydroxy-3-butylphthalide， isopropyl butyrate， L-limonene and 1-methoxyhexane） based on VIP values. HPLC results showed that there was no significant difference of the four components except for ligustilide in wine-processed CR at different stages. ConclusionThis study achieved rapid identification of wine-processed CR with different processing degrees by electronic sensory technology and differential component content detection， with discrimination accuracy rates of 92.4% and 93.272% for color and odor， respectively. This paper also established the reference ranges of main colorimetric parameters for wine-processed CR at different stages， and four differential components were screened out， providing a basis for standardizing the processing of wine-processed CR and establishing quality standards for this decoction pieces.

[Objective] To compare next generation sequencing (NGS) library construction technology between probe hybridization capture and amplicon methods, and analyze the influencing factors of HLA genotyping resolution level and its prospects in clinical applications. [Methods] A total of 207 clinical samples with known typing results and samples from the proficiency testing plan were selected. The conformity rate of HLA genotyping results, allele coverage and typing data analysis indicators were confirmed, and the effects of two library construction methods on the level of HLA genotyping discrimination were compared. [Results] The concordance rate of 207 samples with the feedback results of PT or prior well-characterized HLA genotypes was 100%. Among them, 91 samples were captured using hybridization probe capture method. Compared with the original amplicon method, the hybridization probe capture method can distinguish the alleles of DRB1 and DPB1 that cannot be determined in 13 samples. The allelic imbalance of DRB1, DPA1, and DQB1 loci in 6 samples was resolved. Three samples were found to have missed detection of alleles at the DQA1 and DQB1 loci. [Conclusion] The performance indicators of hybridization probe capture and amplicon performance confirmation meet the requirements of clinical detection of HLA genotyping, which provides an experimental method and basis for clinical application.

Background and aims:Metabolic dysfunction-associated fatty liver disease(MAFLD)is a common chronic condition that can lead to cancer due to its complex pathogenesis.Therapeutic agents targeting AMP-activated protein kinase(AMPK)activation have been suggested as potential treatments for metabolic disorders such as metabolic dysfunction-associated steatohepatitis(MASH).Rhizoma Atractylodis Mac-rocephalae(RAM)has been clinically used to treat obesity-related health problems,but its therapeutic effects on MAFLD and the underlying mechanism remain unclear.Therefore,this study was conducted to evaluate the function and underlying mechanism of RAM in the treatment of MAFLD.Methods:The effect of RAM decoction on MAFLD was evaluated using a high-fat diet(HFD)-induced MAFLD mouse model.In vitro studies were conducted using a palmitic acid/oleic acid-induced lipid accumulation model in the alpha mouse liver 12 cells and RAM-containing serum.The underlying mechanisms were elucidated through a combination of network pharmacology analysis,immunohis-tochemistry,western blotting,and polymerase chain reaction analysis.Results:Administration of RAM decoction significantly reduced body weight gain in MAFLD mice without changing food intake.The weights of the liver and inguinal adipose tissues were also reduced after RAM treatment.Additionally,RAM administration decreased serum levels of alanine aminotrans-ferase,aspartate transaminase,total cholesterol,triglyceride,low-density lipoprotein cholesterol,and glucose,while reducing lipid droplet accumulation in the liver tissues of MAFLD mice.The underlying mechanisms included the activation of the phosphorylation of AMPK and acetyl-CoA carboxylase(ACC),and inhibition of the expression of sterol regulatory element binding protein 1(SREBP1).However,RAM did not alter the protein expression levels of peroxisome proliferator-activated receptor α and carnitine palmitoyltransferase-1α.Furthermore,the RAM-induced upregulation of phosphorylated AMPK,phos-phorylated ACC,and SREBP1 expression,as well as the downregulation of fatty acid synthase expression,were reversed by using an AMPK inhibitor.Conclusions:Through a combination of network pharmacology and experimental validation,we demonstrated that RAM may exert therapeutic effects on MAFLD by inhibiting lipid synthesis and activating phosphorylated AMPK pathways.

Objective To screen antidepressant-active compounds from Polygonati Rhizoma and explore their effects and possible mechanisms against depression induced by simulated weightlessness.Methods A systems pharmacology approach was used to screen potential antidepressant-active compounds and their targets from Polygonati Rhizoma.The hindlimb unloading(HLU)rat model was employed for the study.Twenty-four healthy male Sprague-Dawley rats were randomly divided into three groups:control group(administered 0.5%carboxymethylcellulose by gavage),HLU group(hindlimb unloading),and HLU+treatment group(hindlimb unloading+active compound gavage),with 8 rats in each group.After 28 days of hindlimb unloading,depressive-like behaviors in rats were evaluated using the forced swimming test and tail suspension test.Hippocampal morphology was examined with H&E staining,and GO and KEGG enrichment analyses were conducted on the targets of active compounds.Results A total of 38 active compounds were screened from Polygonati Rhizoma,among which apigenin had an oral bioavailability of 23.06%and a drug-likeness score of 0.21.Compound-target network analysis indicated that apigenin had the highest degree and betweenness centrality values,suggesting it might be the key active component with antidepressant potential in Polygonati Rhizoma.In the forced swimming and tail suspension tests,rats in the HLU group showed a significant increase in immobility time compared to the control group,indicating successful establishment of the depression model.However,compared to the HLU group,rats in the HLU plus apigenin group exhibited significantly reduced immobility time.The H&E staining results of hippocampal tissue showed a significant reduction in the number of hippocampal neurons,along with numerous shrunken neurons and small vacuoles in nerve fibers in the HLU group.In contrast,the treatment group exhibited an increased number of hippocampal neurons,with improved cellular morphology.Target enrichment analysis indicated that apigenin targets were mainly involved in the regulation of apoptosis and cancer-related signaling pathways.Conclusion Apigenin significantly improved depressive-like behaviors in rats subjected to hindlimb unloading,and it has a protective effect on hippocampal tissue.It may provide a new natural active compound for the treatment of depression caused by spaceflight-induced weightlessness.

Objective:To explore the characteristics of depression,anxiety,and stress symptoms among Chinese men who have sex with men(MSM),to determine the links of psychological resilience with these symptoms at the symptom level,and to provide insights for tailoring mental health intervention measures for MSM.Methods:A cross-sectional survey was conducted in southwestern China(Chongqing and Sichuan)from May to August 2022.The Depression,Anxiety,and Stress Scale-21 was used to assess mental health-related symptoms in MSM.Psychological resilience was evaluated using the brief version of the Connor-Davidson Resilience Scale.A regularized partial correlation network was constructed,and then a Bayesian network was established to identify potential causal rela-tionships in symptoms.A flow network was used to explore the link between psychological resilience and symptoms of anxiety,depres-sion,and stress.Results:A total of 938 MSM were included in the analysis.The proportion of MSM with depression,anxiety,and stress was 29.74%."Panic","scared","no relax",and"down-hearted"showed high expected influence.Bridging symptoms were"panic","down-hearted",and"agitated".Central and bridging symptoms also appeared at the top of the Bayesian network.Psychological resil-ience was negatively correlated with"no initiative","down-hearted","meaningless","panic",and"no relax".Conclusion:Central symptoms"panic","scared","no relax",and"down-hearted",as well as bridging symptoms"panic","down-hearted",and"agi-tated"are identified through network analysis.The potential causal priority of these symptoms is prominent.Interventions tailored to central and bridging symptoms may be effective,and interventions for enhancing psychological resilience may alleviate negative emotion-related symptoms,especially depressive symptoms in the MSM population.

Objective To explore the utility of different endoscopic retrograde cholangiopancreatography(ERCP)complexity scales in the training of endoscopists.Methods Clinical data of 237 patients treated by ERCP completed by trainee physicians at two hospitals in Eastern China from February 2022 to February 2023 were prospectively collected.All ERCP cases were classified using previously proposed complexity grading scales,including the American Society of Gastrointestinal Endoscopy(ASGE),Morriston and HOUSE grading scales,compared with the hepatopancreatic ampulla features classification.Successful intubation,successful surgical treatment and complication rates within 48 h were recorded.Results Within each grading,subgroups were divided according to severity,and the comparison of intubation success rates between subgroups in the 3 different grades showed statistically significant differences(P = 0.000).The treatment success rate and intubation success rate had similar findings(P = 0.000).There was no correlation between the complication rate and ASGE grading(P = 0.361),Morriston grading(P = 0.332),and HOUSE grading(P = 0.586).When only cases with primitive papillae were considered,the intubation success rate depended on the complexity grading after the newly added classification of hepatopancreatic ampulla features.In the ASGE group,the success rate of intubation was compared between different subgroups,and the difference was statistically significant(P = 0.004);in the Morriston group,the success rate of intubation was compared between different subgroups,and the success rate of intubation was statistically significant(P = 0.002);in the HOUSE group,the success rate of intubation was compared between different subgroups,and the difference was statistically significant(P = 0.000);In the hepatopancreatic ampulla characteristics group,the difference in intubation success rate was statistically significant when compared between different subgroups(P = 0.000).Similar findings were also found for treatment success rate and intubation success rate(P = 0.000).There was no correlation between the complication rate and ASGE group(P = 0.586),Morriston group(P = 0.443),HOUSE group(P = 0.306),and hepatopancreatic ampulla characteristics group(P = 0.350).Conclusion The use of hepatopancreatic ampulla features as a classification of biliary cannulation complexity is feasible and could be an alternative or additional means of predicting successful biliary cannulation and surgical treatment success and could be used in the future to assess endoscopist training and learning progress and technical standards.

AIM:To investigate the effects of ginkgetin on neurological deficit and angiogenesis induced by cerebral ischemia/reperfusion(I/R)and its underlying mechanism.METHODS:Sixty SD rats were randomly allocated into three groups:sham group,I/R model group,and ginkgetin(40 mg/kg)treatment(I/R+ginkgetin)group,with twenty rats in each group.The middle cerebral artery occlusion/reperfusion(MCAO/R)rat model was employed to simulate cere-bral I/R,and ginkgetin was administered continuously for 7 days following reperfusion.The cerebral infarction volume was quantified using TTC staining.Neuronal density in the ischemic penumbra was assessed through Nissl staining and immu-nohistochemistry for neuron-specific nuclear protein(NeuN).Microvessel density and angiogenesis in the ischemic pen-umbra of each group were analyzed using CD31 labeling and BrdU/von willebrand factor(vWF)double labeling immuno-fluorescence staining.Western blot analysis was performed to determine the levels of heat shock protein 70(HSP70),vas-cular endothelial growth factor(VEGF),and hypoxia inducible factor-1α(HIF-1α)in the ischemic penumbra.RE-SULTS:Compared with the I/R model group,the cerebral infarction volume was significantly reduced in ginkgetin treat-ment group(P<0.01),the number of neurons,the microvessel density,angiogenesis and the expression levels of HIF-1α,VEGF,and HSP70 in the ischemic penumbra were significantly increased(P<0.01).CONCLUSION:Ginkgetin exhibits the potential to augment angiogenesis and facilitate neurological function recovery in MCAO rats,while concur-rently upregulating the expression of HSP70,VEGF,and HIF-1α within the ischemic penumbra.

Objective To study the predictive value of features of the hepatopancreatic ampulla on the success of endoscopic retrograde cholangiopancreatography(ERCP)cannulation and complication rates.Methods Clinical data of 400 patients who underwent ERCP from June 2023 to October 2023 at two hospitals in Eastern China were retrospectively analyzed,patients were divided into non-protruding group(n=184),protruding group(n=76),diverticulum,mucosal folds group(n=101)and twisted,tumour involved group(n=39)according to hepatopancreatic ampulla features.The frequency of nipple contact during intubation,time to successful intubation,and procedure-related complications were recorded separately.Results The highest mean number of contacts in the nipple contact frequency was found in the twisted,tumour-involved group(8.95±6.30)times,and the lowest number of contacts was found in the non-protruding group(4.01±2.42)times,the difference was statistically significant in four groups(F=31.06,P=0.000).Hepatopancreatic ampulla features were a significant factor influencing cannulation time,with prolonged cannulation time observed in the twisted,tumour-involved group(353.96±263.42)s and fastest cannulation in the non-protruding group(161.03±118.06)s,the difference was statistically significant in four groups(F=17.73,P=0.000).A total of 31 patients(7.75%)experienced complications,with the highest number of post-ERCP pancreatitis and no cases of perforation or death.Conclusion Hepatopancreatic ampulla features are a simple and feasible way to predict the success of ERCP cannulation and the incidence of complications.In the future,it could be used as a biological predictor of the difficulty of bile duct cannulation.

Objective:To establish a linkage prediction model for human leukocyte antigen (HLA) DPA1-DPB1 and validate it by using clinical data and follow-up data from unrelated allogeneic hematopoietic stem cell transplantation donors and recipients, and to explore the clinical application value of the prediction model in transplantation prognosis.Methods:This is a retrospective study. Leveraging the artificial neural network algorithm of NetMHCⅡpan and the DPA1-DPB1 haplotype linkage database of the Chinese population established in our previous research, and incorporating the amino acid FASTA data of DPA1-DPB1 of all known sequences newly published by the Latest International Immunogenetics/Human Leukocyte Antigens, 47 DPA1-DPB1 linkage models were established. Employing next-generation sequencing technology based on the hybridization capture library construction method, HLA genotyping tests for HLA-A, -B, -C, DRB1, DQB1, DQA1, DRB3/4/5, DPB1, and DPA1 (9 loci) were performed on 250 donor-recipients pairs who underwent unrelated-donor hematopoietic stem cell transplantation in the Department of Hematology of the First Affiliated Hospital of Soochow University between January 2016 and September 2021. HLA typing data and clinical information of transplant donors and recipients were retrospectively analyzed to assess and predict the impact of permissive and non-permissive linkage mismatches of DPA1-DPB1 on transplantation prognosis. The Kaplan-Meier method with the log-rank test was applied to compare the survival curves of overall survival (OS) rates between different groups. Additionally, a competing risks model was utilized to compare the cumulative incidence of grade Ⅱ-Ⅳ acute graft-versus-host disease and non-relapse mortality (NRM) across groups. The area under the receiver operating characteristic curve was employed to compare the predictive performance of the established prediction model with that of the T-cell epitope (TCE) model.Results:According to the different hydrophilic and hydrophobic properties of amino acids, the DPA1-DPB1 linkage model is categorized into types Ⅰ-Ⅳ: type I consists of 6 hydrophobic types at P1-P8 plus hydrophilic type at P9; type Ⅱ includes 17 hydrophobic types; type Ⅲ comprises 9 amphiphilic types; and type Ⅳ consists of 15 hydrophilic types. According to the prediction model, DPA1-matched and DPB1-mismatched donor-recipient cases were classed into P1-matched or P1-mismatched groups. Compared with fully matched DPA1 and DPB1 cases, P1-mismatched patients had a 2-year OS rate of 75% (12/16) versus 96.2%(25/26) (χ2=4.13, P=0.04), and a NRM rate of 4/16 versus 0 (χ2=7.05, P<0.01). However, there was no statistically significant difference in the 2-year OS and NRM rates compared to DPA1 and DPB1 cases ( P>0.05). The prediction model established in this study demonstrated a larger area under the receiver operating characteristic curve for predicting the 2-year OS rate compared with the DPB1 TCE model ( Z=0.71, P=0.48). In donor-recipient cases where both DPA1 and DPB1 were mismatched, the 2-year OS rates decreased and the NRM increased in both P1-matched and P1-mismatched cases compared with fully matched DPA1 and DPB1. Moreover, P1-mismatched patients had a worse prognosis compared to P1-matched patients. Conclusion:The DPA1-DPB1 linkage prediction model established based on high-throughput next-generation sequencing technology can be used to predict the impact of HLA-DP mismatches on OS and NRM in transplantation, and the prediction performance is superior to the TCE model.