1.Characteristics of asymptomatic HBV infection in HBsAg-positive blood donors from Dalian
Yingying WANG ; Xuelian DENG ; Xiaohan GUO ; Huihui GAO ; Peng SUN ; Dan LIU ; Daniel CANDOTTI ; Bing WANG
Chinese Journal of Blood Transfusion 2025;38(10):1361-1369
Objective: To analyze serological and molecular characteristics of asymptomatic HBV infection in HBV surface antigen positive (HBsAg+) blood donors from Dalian. Methods: The prevalence of HBsAg was analyzed among blood donors in Dalian between 2013 and 2022. Randomly selected HBsAg+ blood samples were subjected to HBV serological testing, HBV viral DNA quantification, and HBV genotyping. Results: Over this ten-year period, the prevalence of HBsAg decreased from 1.25% to 0.50% among blood donors in Dalian. Donors who tested positive for HBsAg prior to donation using a rapid test (RT) accounted for 92.5% of all HBsAg+ donors identified. A total of 240 confirmed HBsAg+ blood donors were randomly selected, including 125 donors with positive results and 115 with negative results in the pre-donation rapid test. HBsAg+ donors were mainly males (71.2%), with a median age of 42, and 97.5% of them being first-time donors. Based on HBV serological profiles, three stages of infection were identified: early infection (2.9%), suspected acute hepatitis (0.8%), and chronic infection (96.3%). The dominant HBV genotypes were C (68.9%) and B (28.4%). Among chronic HBV infection individuals, donors infected with HBV genotype B were older than those infected with genotype C (median age: 45y vs 38.5y, P<0.05). Additionally, they showed significantly lower HBsAg levels with a narrower distribution range than those infected with genotype C [median: 23.2 IU/mL (range: <0.05-7 910 IU/mL) vs 968 IU/mL (range: <0.05-3.4×10
), P<0.05]. However, no significant difference was observed in the HBV DNA loads between these two genotypes (P>0.05). Conclusion: Between 2013 and 2022, the prevalence of HBsAg among blood donors in Dalian showed a year-over-year decline. Chronic infection was predominant among HBsAg+ first-time blood donors. The characteristics of chronic infection in blood donors differed significantly depending on the viral genotype, manifesting as differences in age of infected individuals and HBsAg level distribution.
2.Dual effects of branched-chain amino acid on differentiation of 3T3-L1 preadipocytes through Stat3 pathway
Xinghua Cai ; Jie Gao ; Yuanying Xu ; Huihui Zhang ; Rouzi Maireyanmu ; Wenjun Sha ; Jun Lu ; Tao Lei
Acta Universitatis Medicinalis Anhui 2025;60(3):494-501
Objective :
To investigate the effects of branched-chain amino acid(BCAA) on the differentiation of 3T3-L1 preadipocytes and its potential mechanism.
Methods :
3T3-L1 preadipocytes were divided into the Control, differentiation medium(DM), low-concentration BCAA, and high-concentration BCAA groups. A CCK-8 assay was utilized to evaluate pre-adipocyte survival under various BCAA concentrations. Oil-red O staining was used to observe the formation of lipid droplets in adipocytes. Intracellular triglyceride(TG) and total cholesterol(TC) were detected by enzymatic method. RT-qPCR and Western blot were used to detect the mRNA and protein expression of Stat3 and adipocyte differentiation-related genes.
Results :
CCK-8 results showed that the viability of 3T3-L1 cells was not affected when the BCAA concentration was ≤ 10 mmol/L. Compared with the DM group, the low-concentration BCAA groups(0.5 and 1.0 mmol/L) had significantly larger intracellular lipid droplets, increased number of lipid droplets, and elevated levels of the intracellular TC(0.88vs0.68 mmol/g; 0.83vs0.68 mmol/g,P<0.01) and TG(0.77vs0.40 mmol/g; 0.62vs0.40 mmol/g,P<0.01). Nevertheless, the cell differentiation in the high-concentration group(5.0 and 10.0 mmol/L) significantly decreased compared with that in the DM group. Further, levels of PPARγ, C/EBPα, Adiponectin, and FABP4 mRNA and protein expression significantly increased in the low-concentration group, but significantly decreased in the high-concentration group than that in the DM group(P<0.01). In addition, low concentrations of BCAA promoted stat3 phosphorylation, while high concentrations inhibited its phosphorylation(P<0.01).
Conclusion
BCAA have a dual role in regulating the differentiation of preadipocytes through Stat3, i.e. low concentrations of BCAA induce cell differentiation by promoting Stat3 phosphorylation; whereas high concentrations of BCAA inhibit Stat3 phosphorylation and cell differentiation.
3.Paeoniflorin alleviates LPS-induced aerobic glycolysis in HK-2 cells by modulating the PI3K/AKT/HIF-1α pathway
Jing XIE ; Ruonan LI ; Huihui GAO ; Shunkai YANG ; Yuqing MA
Journal of Army Medical University 2025;47(20):2483-2494
Objective To investigate the effects of paeoniflorin(PF)on lipopolysaccharide(LPS)-induced aerobic glycolysis in renal tubular epithelial cell line HK-2 and its underlying mechanism of action.Methods This study consists of a preliminary experiment and a formal experiment.Preliminary experiment:CCK-8 assay and RT-qPCR were used respectively to measure cell viability and mRNA expression levels of inflammatory factors in HK-2 cells after LPS stimulation to determine the optimal LPS concentration for modeling as well as to evaluate the toxicity of PF and screen for its appropriate concentration.Formal experiment:HK-2 cells were divided into control group(CON group),LPS group,LPS+PF group and LPS+PF+740Y-P group.LPS was used to establish a cell model of sepsis associated-acute kidney injury(SA-AKI)in HK-2 cells,and then the cell model was treated with PF and PI3K activator 740Y-P,correspondingly for 24 h.CCK-8 assay was employed to detect cell viability,and Extracellular Acidification Rate(ECAR)Kit was utilized to measure the rate.The contents of IL-1β,IL-18,lactic acid(Lac)and lactate dehydrogenase A(LDHA)were determined with ELISA.Western blotting was applied to detect the expression of p-PI3K,p-AKT,HIF-1α,pyruvate kinase 2(PKM2,a key enzyme in aerobic glycolysis)and NOD-like receptor thermal protein domain associated protein 3(NLRP3),and immunofluorescence assay was performed to observe the expression and distribution of PKM2.Results ① Our preliminary experiment identified that the optimal concentration of LPS for modeling was 20.0 μg/mL,a safe dosage range of PF was 0~100.0 μmol/L,and its optimal therapeutic concentration was 25.0 μmol/L.② Compared with the CON group,LPS stimulation resulted in significantly decreased cell viability(P<0.05),increased ECAR(P<0.05),elevated contents of IL-1β,IL-18,Lac and LDHA(P<0.05),up-regulated protein levels of p-PI3K,p-AKT,HIF-1α,p-PKM2 and NLRP3(P<0.05),and enhanced fluorescence intensity of PKM2 in the nucleus of cells(P<0.05).Compared with the model group,PF treatment reversed all above effects induced by LPS stimulation(all P<0.05).Compared with the LPS+PF group,in the LPS+PF+740Y-P group,ECAR was elevated(P<0.05),the contents of IL-1β,IL-18,Lac and LDHA were increased(P<0.05),and the relative expression levels of p-PI3K,p-AKT,HIF-1α,p-PKM2 and NLRP3 were increased(P<0.05),and the fluorescence intensity of PKM2 was strengthened(P<0.05)and enhanced in the nucleus(P<0.05).Conclusion PF reduces aerobic glycolysis in HK-2 cells and alleviates the inflammatory response by inhibiting the PI3K/AKT/HIF-1α signaling pathway.
4.Construction and validation of prediction models for delayed encephalopathy after acute carbon monoxide poisoning based on machine learning
Yanwu YU ; Yan ZHANG ; Ding YUAN ; Huihui HAO ; Fang YANG ; Hongyi YAN ; Pin JIANG ; Mengnan GUO ; Zhigao XU ; Changhua SUN ; Gaiqin YAN ; Lu CHE ; Jianjun GUO ; Jihong CHEN ; Yan LI ; Yanxia GAO
Chinese Journal of Emergency Medicine 2025;34(10):1403-1409
Objective:s To investigate the risk factors for delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) in patients with acute carbon monoxide poisoning (ACOP) and to develop predictive models based on machine learning algorithms.Methods:Patients with ACOP hospitalized at the First Affiliated Hospital of Zhengzhou University from August 2019 to October 2024 were included, with the occurrence of DEACMP as the outcome measure. The dataset was randomly divided into training and validation sets at a ratio of 7:3. Lasso regression was used to select features influencing the outcome in training sets. Nine machine learning models—including Random Forest (RF), Extreme Gradient Boosting (XGBoost), and Support Vector Machine (SVM)—were constructed. Receiver operating characteristic (ROC) curves were plotted and the area under the curve (AUC) calculated for each model. Calibration curves were used to assess accuracy, and decision curve analysis (DCA) was applied to evaluate clinical utility. The SHapley Additive exPlanations (SHAP) method was employed to visualize and interpret the best-performing model.Results:A total of 264 ACOP patients were included, of whom 54 (20.5%) developed DEACMP. Lasso regression identified eight key feature variables. Based on these factors, predictive models were constructed, showing good AUC stability across the nine machine learning models in both training (0.92–0.99) and validation sets (0.85–0.91). The RF model performed best, with an AUC of 0.99 in the training set and 0.90 in the validation set; its calibration curve and DCA curve also demonstrated excellent performance. SHAP analysis of the RF model revealed the importance ranking of factors from highest to lowest as follows: Glasgow Coma Scale (GCS) score, duration of coma, age, history of coronary heart disease, CK-MB level, monocyte count, diastolic blood pressure (DBP), and drinking history.Conclusions:The RF model exhibited the highest predictive performance for DEACMP occurrence in ACOP patients. The influencing factors, ranked in order of importance from highest to lowest, are as follows: GCS score, duration of coma, age, history of coronary heart disease, CK-MB level, monocyte count, DBP, and drinking history.
5.Correlation between blood microRNA-133b and soluble FMS-like tyrosine kinase 1 levels and prognosis in patients with endometrial cancer
Huihui SUN ; Yanjuan GUO ; Nannan ZHAO ; Jianli ZHOU ; Jinling YUAN ; Jie GAO
Chongqing Medicine 2024;53(19):2943-2948
Objective To study the relationship between blood microRNA-133b(miR-133b)and solu-ble fms-like tyrosine kinase 1(sFLT1)levels with the prognosis in the patients with endometrial cancer.Methods A total of 122 patients with endometrial cancer visited in the gynecology department of this hospital from January 2015 to January 2016 were prospectively selected as the study subjects,and divided into the sur-vival group(n=58)and death group(n=64)according to the 5-year prognosis of the patients with endome-trial cancer.The miR-133b and sFLT1 levels were compared between the two groups.The COX regression was used to analyze the relationship between miR-133b and sFLT1 with the prognosis of the patients with en-dometrial cancer.Results The levels of miR-133b and sFLT1 in the survival group were higher than those in the death group,and the differences were statistically significant(P<0.05).The median survival time in the miR-133b low-level group was shorter than that in the miR-133b high level group,and the difference was sta-tistically significant(P<0.05).The median survival time of the sFLT1 low level group was shoeter than that in the sFLT1 high level group,and the difference was statistically significant(P<0.05).The FIGO stageⅢ-Ⅳ and lymph node metastasis were the independent risk factors for the prognosis of endometrial cancer(P<0.05),and the pathological G1-G2,high level of miR-133b and sFLT1 were the independent protective factors for the prognosis of endometrial cancer(P<0.05).Conclusion The miR-133b and sFLTl low levels in the patients with endometrial cancer are associated with the disease progression,and both are the independ-ent risk factors of prognosis.
6.Computer-aided diagnosis for prostate cancer detection in multiparametric MRI:a multireader multicase study
Ge GAO ; Kexin WANG ; Huihui WANG ; Rui WANG ; Yi LIU ; Xiaoying WANG
Journal of Practical Radiology 2024;40(5):825-829
Objective To investigate the effect of computer-aided diagnosis(CAD)as a"second reader"integrated in prostate cancer(PCa)multiparametric magnetic resonance imaging(mpMRI)on the diagnostic performance.Methods Sixty-four patients were recruited retrospectively.Six readers were divided into 3 groups according to the experience.Unknown the pathologic diagnosis,readers graded according to prostate imaging reporting and data system(PI-RADS)v2.1 and accepted or rejected the revision of PI-RADS v2.1 scores by CAD.Using pathology as the gold standard,the diagnostic efficiency,diagnostic confidence and reading time of PI-RADS v2.1 were compared before and after CAD assistant.Results On the level of lesions,without CAD,the area under the curve(AUC)of reader A1 was higher than B1,C1 and C2,and the AUC of reader A2 was higher than C1 and C2,the difference was statistically significant(P<0.05).While the difference of AUC among other readers were not significant(P>0.05).With CAD,the difference of AUC among all readers were not significant(P>0.05).On the level of patients,the mean AUC of all readers with CAD[0.903,95%confidence interval(CI)0.848-0.958]was higher than that without CAD(0.863,95%CI 0.800-0.929),the differ-ence was statistically significant(F=6.52,P=0.040).The difference of AUC between without and with CAD assistance was-0.040(95%CI-0.078--0.003),standard deviation was 0.016,the difference was statistically significant(t=-2.55,P=0.040).The diag-nostic confidence of readers was improved significantly with CAD(P<0.05).Besides,the average reading time of each case required an additional 0.8 min.Conclusion After integrating CAD into PCa mpMRI diagnosis as a"second reader",the performance of less experienced readers can be improved and show similar diagnostic efficiency with experienced readers.
7.Hypericin reduces myocardial ischemia-reperfusion injury in rats by modulating AMPK/Nrf2/HO-1 signaling pathway
Huihui LI ; ANWER GULINIGAER ; Xiaofeng GAO ; Gang WANG ; Yali BAO ; Tian ZHANG ; AINIWAR DINA ; Can LING ; Zhan SUN
Chinese Journal of Pathophysiology 2024;40(10):1882-1890
AIM:To investigate the effect of hypericin(Hyp)on the heart of rats with myocardial ischemia-re-perfusion injury(MIRI),and to explore its mechanism.METHODS:Thirty SPF male SD rats were divided into 5 groups:sham group,MIRI group,low-dose Hyp(L-Hyp)group(MIRI+L-Hyp group),high-dose Hyp(H-Hyp)group(MIRI+H-Hyp group),and positive control trimetazidine(TMZ)group(MIRI+TMZ group),with 6 rats in each group.Apart from the sham group,the MIRI model was established by ligating the anterior descending branch of the left coronary artery and then recanalizing it in the remaining four groups of rats.The success of the modeling was determined by monitor-ing the electrocardiogram.We assessed the cardiac function in rats using echocardiography.TTC staining was employed to measure the area of myocardial infarction in rats,and HE staining was utilized to observe the morphological traits of rat myocardium.We assayed the activities of lactate dehydrogenase(LDH)and superoxide dismutase(SOD)and the levels of malondialdehyde(MDA)in rat serum using biochemical kits.ELISA kits were applied to assess the contents of cardiac troponin I(cTnI)and reactive oxygen species(ROS)in rat serum.Western blot analysis was perfomed to measure the pro-tein expression levels of AMPK,p-AMPK,Nrf2,and HO-1 in rat myocardial tissues.RESULTS:The rats in MIRI group exhibited increased myocardial tissue injury,larger myocardial infarction areas,decreased left ventricular ejection fraction(LVEF),and reduced left ventricular fractional shortening(LVFS)compared with sham group,as shown by echo-cardiography.Additionally,there were increases in LDH activity,cTnI,MDA and ROS levels,along with significant de-creases in SOD activity,and p-AMPK,Nrf2 and HO-1 protein levels(P<0.05).Compared with MIRI group,the rats in MIRI+L-Hyp,MIRI+H-Hyp and MIRI+TMZ groups showed decreased myocardial histopathological damage and reduced myocardial tissue infarction area,increased LVEF and LVFS,and lowered serum levels of LDH activity,cTnI,MDA and ROS,while SOD activity,p-AMPK,Nrf2 and HO-1 protein levels were elevated(P<0.05).CONCLUSION:Hypericin attenuates myocardial ischemia-reperfusion injury in rats,possibly by modulating the AMPK/Nrf2/HO-1 signaling path-way.
8.Establishment of predictive model for post-induction hypotension in patients undergoing colorectal cancer resection based on muscle CT parameters: machine learning algorithms
Weixuan SHENG ; Danyang GAO ; Huihui MIAO ; Tianzuo LI
Chinese Journal of Anesthesiology 2024;44(11):1293-1299
Objective:To establish a predictive model for post-induction hypotension (PIH) in the patients undergoing colorectal cancer resection using machine learning algorithms based on muscle CT parameters.Methods:This was a single-center, retrospective study. Electronic medical records from 318 patients who underwent colorectal cancer resection from September 1, 2018 to September 30, 2021 at our hospital were collected. Predictive variables included age, gender, body mass index, hemoglobin, American Society of Anesthesiologists Physical Status classification, TNM staging, age-adjusted Charlson comorbidity index, prognostic nutritional index, L 3 level skeletal muscle index, and muscle quality assessed by Hounsfield unit average calculation. The outcome variable was PIH. The training and testing sets were divided based on the timeline (patients before September 1, 2020 were included in the training set, and those after that date were included in the testing set). The filtering method was used to screen the feature variables. Eight models, including logistic regression, Bayesian models, K-nearest neighbors, support vector machines, neural networks, decision trees, extreme gradient boosting trees, and random forests, were established in the training set using over-sampling technique, repeated cross-validation and hyperparameter optimization. After selecting the best model, a sorting chart of the feature variables, a univariate partial dependency profile, and a breakdown profile were drawn. In the testing set, the confusion matrix and parameters were calculated, and the receiver operating characteristic curve, precision recall curve, calibration curve, and decision curve were drawn to evaluate the performance of the predictive model. Results:The screened feature variables were Hounsfield unit average calculation value, age, L 3 level skeletal muscle index, prognostic nutritional index, hemoglobin and body mass index. The random forest was the optimal model, with an accuracy of 0.985 9, a MCC of 0.970 8, the area under the receiver operating characteristic curve was 1.0, and the area under the precision recall curve was 1.0. The Brier score of the calibration curve was 0.076 6; the decision curve showed the highest clinical net benefit of 0.6. Conclusions:In this study, machine learning algorithm is used to identify the important characteristic variables and establish a high-performance PIH prediction model based on muscle CT parameters.
9.Research Progress in the Pathogenesis of Polycystic Ovary Syndrome
Huihui GAO ; Beiran QIAN ; Yan NI ; Liying SUN ; Junfen FU
Journal of Sichuan University (Medical Sciences) 2024;55(4):1049-1054
Polycystic ovary syndrome(PCOS)is one of the most common gynecological endocrine disorders.Most pathophysiological changes of PCOS begin in the peripubertal phase,and these pathophysiological changes will continuously affect women's health in the later stages of their lives.The pathogenic mechanisms of PCOS remain unclear,involving key aspects such as the regulation of hypothalamic-pituitary function,ovarian cellular functions,androgen levels,and insulin resistance.Herein,we summarized the latest findings on the pathogenesis of PCOS from the perspectives of the genetic background,intrauterine development,neuroendocrine function,inflammatory factors,gut microbiome,and environmental factors.This review will help provide new ideas for a deeper understanding of the disease,as well as its clinical diagnosis and treatment.
10.Comparative analysis of differentially expressed genes for biosynthesis of active ingredients in fruits of different cultivars of Lycium barbarum L. based on transcriptome sequencing.
Xuexia LIU ; Wenqiang FAN ; Huihui JIAO ; Han GAO ; Jianning TANG ; Jinzhong ZHU ; Sijun YUE ; Rui ZHENG
Chinese Journal of Biotechnology 2023;39(7):3015-3036
To explore the differentially expressed genes (DEGs) related to biosynthesis of active ingredients in wolfberry fruits of different varieties of Lycium barbarum L. and reveal the molecular mechanism of the differences of active ingredients, we utilized Illumina NovaSeq 6000 high-throughput sequencing technology to conduct transcriptome sequencing on the fruits of 'Ningqi No.1' and 'Ningqi No.7' during the green fruit stage, color turning stage and maturity stage. Subsequently, we compared the profiles of related gene expression in the fruits of the two varieties at different development stages. The results showed that a total of 811 818 178 clean reads were obtained, resulting in 121.76 Gb of valid data. There were 2 827, 2 552 and 2 311 DEGs obtained during the green fruit stage, color turning stage and maturity stage of 'Ningqi No. 1' and 'Ningqi No. 7', respectively, among which 2 153, 2 050 and 1 825 genes were annotated in six databases, including gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) and clusters of orthologous groups of proteins (KOG). In GO database, 1 307, 865 and 624 DEGs of green fruit stage, color turning stage and maturity stage were found to be enriched in biological processes, cell components and molecular functions, respectively. In the KEGG database, the DEGs at three developmental stages were mainly concentrated in metabolic pathways, biosynthesis of secondary metabolites and plant-pathogen interaction. In KOG database, 1 775, 1 751 and 1 541 DEGs were annotated at three developmental stages, respectively. Searching the annotated genes against the PubMed database revealed 18, 26 and 24 DEGs related to the synthesis of active ingredients were mined at the green fruit stage, color turning stage and maturity stage, respectively. These genes are involved in carotenoid, flavonoid, terpenoid, alkaloid, vitamin metabolic pathways, etc. Seven DEGs were verified by RT-qPCR, which showed consistent results with transcriptome sequencing. This study provides preliminary evidences for the differences in the content of active ingredients in different Lycium barbarum L. varieties from the transcriptional level. These evidences may facilitate further exploring the key genes for active ingredients biosynthesis in Lycium barbarum L. and analyzing their expression regulation mechanism.
Flavonoids/metabolism*
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Fruit/genetics*
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Gene Expression Profiling/methods*
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Gene Expression Regulation, Plant
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Lycium/metabolism*
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Metabolic Networks and Pathways
;
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