The antidepressant activity and molecular mechanisms of Yueju Wan volatile oil were investigated. The Yueju Wan volatile oil was extracted by using supercritical CO_2. Gas chromatography-mass spectrometry(GC-MS) combined with network pharmacology identified 28 chemical constituents in Yueju Wan volatile oil, primarily terpenes and lactones. A total of 123 overlapping targets were associated with depression, including core targets of interleukin-1β(IL-1β), signal transducer and activator of transcription 3(STAT3), and caspase-3(CASP3). These targets were mainly involved in the prolactin, advanced glycation end products/receptor(AGE/RAGE), and phosphoinositide 3-kinase/protein kinase B(PI3K/Akt) signaling pathways. A reserpine-induced depression mouse model was established to evaluate the therapeutic effects and mechanisms of Yueju Wan volatile oil. The effects of Yueju Wan volatile oil on depression-like behavior in mice were evaluated by analyzing body mass, body temperature index, tail suspension immobility time, forced swimming immobility time, and sucrose preference. Hematoxylin-eosin(HE) staining revealed neuronal protection of Yueju Wan volatile oil in the brain of mice. Enzyme-linked immunosorbent assay(ELISA) and Western blot were employed to detect the protein expression of AGEs, IL-1β, phosphorylated PI3K(p-PI3K), Akt, phosphorylated Akt(p-Akt), nuclear factor κB(NF-κB), and brain-derived neurotrophic factor(BDNF). Behavioral evaluation showed that Yueju Wan volatile oil could effectively control the decline of body mass and body temperature of depressed mice, reduce tail suspension and swimming immobility time, and enhance their preference for sucrose. Histopathological examination showed that Yueju Wan volatile oil could alleviate the neuronal damage in CA1 and dentate gyrus(DG) of the hippocampus of mice. ELISA and Western blot results showed that Yueju Wan volatile oil could significantly increase the protein expression levels of PI3K, Akt, and BDNF and significantly decrease the protein expression levels of AGEs, IL-1β, p-PI3K, p-Akt, and NF-κB in the hippocampus of mice. Furthermore, the p-PI3K/PI3K and p-Akt/Akt ratios were significantly decreased at medium and high doses. These findings suggest that the aromatherapy of Yueju Wan volatile oil can significantly improve reserpine-induced depression-like behavior in mice, which may be related to reducing the expression of neuronal membrane protein AGEs, reducing the phosphorylation levels of PI3K and Akt, inhibiting NF-κB entry into the nucleus, and alleviating the release of pro-inflammatory factors and nerve injury.
Animals ; Antidepressive Agents/chemistry* ; Mice ; Proto-Oncogene Proteins c-akt/immunology* ; Phosphatidylinositol 3-Kinases/immunology* ; Oils, Volatile/chemistry* ; Male ; Drugs, Chinese Herbal/chemistry* ; Signal Transduction/drug effects* ; Depression/metabolism* ; Glycation End Products, Advanced/immunology* ; Humans

Myocardial infarction is one of the severe cardiovascular diseases.The patients with myocardial infarction die of heart failure or arrhythmia.In recent years,the studies in myocardial infarction therapies have advanced greatly,especially the preclinical experimental studies.The experimental studies of myocardial infarction often rely on animal models.Therefore,successful establishment of the myocardial infarction models has important application value in exploring the new techniques and measures for repairing the infarcted myocardium.In this paper,the techniques in establishment of the myocardial infarction models and strategies of their application are summarized.

Objective:To develop and validate a postoperative infection nomogram of hepatitis B-associated hepatocellular carcinoma (HCC) after hepatectomy.Methods:Clinical data of 229 patients with HCC undergoing hepatectomy at the Department of Hepatobiliary Surgery of Tianjin Third Central Hospital from January 2014 to December 2022 were retrospectively analyzed, including 174 males and 55 females, aged (58.2±11.4) years. LASSO regression analysis screened the factors associated with hepatitis B-associated HCC infection after hepatectomy, which were further incorporated into multivariate logistic regression analysis. A nomographic prediction model was established based on the results of multivariate logistic regression analysis. Concordance index (C-index), calibration curve and receiver operating characteristic (ROC) curve were used to evaluate the model, and decision curve analysis (DCA) was used to analyze the clinical applicability of the model. Internal validation of the model was performed using bootstrap method.Results:A total of nine variables were screened as factors associated with the postoperative infections using LASSO regression, including gender, smoking history, body mass index (BMI), serum level of alpha fetoprotein, resection fashion (anatomical or non-anatomical), intraoperative blood loss, surgical method (laparoscopy or open), serum level of creatinine, and postoperative biliary fistula. Multivariate logistic regression analysis showed that BMI, resection fashion, intraoperative blood loss >500 ml, and postoperative biliary fistula were risk factors for postoperative infection (all P<0.05). Based on the above risk factors, a postoperative infection nomogram of hepatitis B-associated HCC after hepatectomy was established. The C-index was 0.839 (95% CI: 0.768-0.910), and the area under ROC curve was 0.853 (95% CI: 0.795-0.912), indicating that the model had a good predictive ability. The calibration curve was basically consistent with the ideal curve. The DCA showed that the model had a good clinical applicability. Internal validation C-index was 0.829 (95% CI: 0.766-0.892). Conclusion:The nomogram based on BMI, surgical resection fashion, intraoperative blood loss >500 ml, and postoperative biliary fistula has a high predictive accuracy and can be used to predict postoperative infections after hepatectomy for HCC.

Objective:To explore the effect of TP53 mutation variant allele frequency(VAF)on the prognosis of diffuse large B-cell lymphoma(DLBCL)patients.Methods:This study included 155 patients with DLBCL who were first diagnosed in the People's Hospital of Xinjiang Uygur Autonomous Region from March 2009 to March 2022. Complete clinical data and paraffin-embedded tumor tissue samples were obtained,and DNA was extracted from tumor tissues.The gene mutation profile of DLBCL patients was detected and analyzed by second-generation sequencing technology.Kaplan-Meier method was used to analyze the mutation status of TP53 gene and the relationship between mutation VAF and OS.Cox regression univariate and multivariate analysis was use to analyze the independent factors affecting OS.A nornogram model for predicting 1,3,and 5 years OS in DLBCL patients were established to evaluated the performance of the model based on C-index and calibration curves.Results:The average value of TP53 mutation VAF in male DLBCL patients was significantly higher than that in female patients (P<0.05 ).Patients with TP53 mutantion had shorter OS than those with wild-type patients (P=0.030).The optimal VAF threshold for TP53 mutation based on OS stratification was 33.61％(P<0.001),and patients with TP53 mutation VAF≥34％had shorter OS than those with TP53 mutation VAF<34％and wild-type patients (P<0.001).Multivariate Cox analysis showed that TP53 mutation VAF≥34％ was an independent poor predictor of OS (HR=4.05,P<0.001),and IPI score ≥3 was an independent predictor of OS poor (HR=2.27,P=0.008).In combination with factors with independent prognostic significance obtained from multi-factor analysis,we constructed a nomogram model for predicting 1-year,3-year,5-year OS in DLBCL patients.The results showed that the C index of TP53-mutated VAF combined with IPI model was 0.743,which predicted the value of 1-year,3-year,and 5-year OS in DLBCL patients.Calibration curves show that the model has good agreement between predicted and actual survival of DLBCL patients at 1-year,3-year,and 5-year. Conclusion:TP53 mutant VAF has prognostic value in DLBCL patients,and TP53 mutant VAF≥34％ is an independent risk factor for OS in DLBCL patients.The prognosis model of TP53 mutation VAF combined with IPI nomogram constructed in this study has good predictive performance for DLBCL patients.

Efficient bacterial drug susceptibility test can help precise guidance on using antimicrobial and improve the efficiency of development of antibiotics.Traditional antimicrobial susceptibility testing(AST)methods are simple,reliable and specific,but suffer from drawbacks of time-consuming.The CCK8 reagent can be reduced by the dehydrogenase in the bacterial cells to form a water-soluble orange-yellow formazan,and the concentration of viable bacteria is proportional to the absorbance of formazan at 450 nm(OD450 nm).In this work,based on the principle of CCK8 colorimetry,the antibiotic loaded composite fiber membrane was used as the culture matrix for AST.Simple,rapid and high-throughput AST was realized by the difference in OD450 nm,which reflected the differences in viable bacteria quantity during bacterial growth.The total detection time was less than 8 h.The potential application of the method was evaluated using S.aureus,P.aeruginosa in wound simulation fluid and E.coli in artificial urine as simulated actual samples.The results of this method were in consistent with the disk diffusion test.This method provided a new idea for AST,which was helpful for the guidance of clinical medication and the development of antibiotics.

OBJECTIVES: To evaluate the performance of different multi-modality fusion models for predicting radiation-induced oral mucositis (RIOM) following radiotherapy in patients with nasopharyngeal carcinoma (NPC). METHODS: We retrospectively collected the data from 198 patients with locally advanced NPC who experienced RIOM following radiotherapy at the Affiliated Tumor Hospital of Guangzhou Medical University from September, 2022 to February, 2023. Based on oral radiation dose-volume parameters and clinical features of NPC, basic classification models were developed using different combinations of feature selection algorithms and classifiers and integrated using a multi-criterion decision-making (MCDM)-based classifier fusion (MCF) strategy and its variant, the H-MCF model. The basic classification models, MCF model, the H-MCF model with a single modality or multiple modalities and other ensemble classifiers were compared for performances for predicting RIOM by assessing the area under the ROC curve (AUC), accuracy, sensitivity, and specificity. RESULTS: The H-MCF model, which integrated multi-modality features, achieved the highest accuracy for predicting severe RIOM with an AUC of 0.883, accuracy of 0.850, sensitivity of 0.933, and specificity of 0.800. CONCLUSIONS Compared with each of the individual classifiers, the multimodal multi-classifier fusion algorithm combining clinical and dosimetric modalities demonstrates superior performance in predicting the incidence of severe RIOM in NPC patients following radiotherapy.
Humans ; Nasopharyngeal Carcinoma/radiotherapy* ; Nasopharyngeal Neoplasms/radiotherapy* ; Retrospective Studies ; Stomatitis/diagnosis* ; Algorithms ; Radiation Injuries/diagnosis* ; Female ; Male ; ROC Curve

Objective:To analyze the detection strategy and basic detection situation of markers of infectious diseases transmitted by transfusion in blood testing laboratories of blood stations in China.Methods:Based on the data of practice comparison working party of Blood Stations in Mainland of China from 2017 to 2021, the data on the testing strategies and the basic detection information of the markers for the transmission of infectious diseases through transfusion in the member laboratories of the practice comparison working party of Blood Stations in Mainland of China from 2017 to 2021 were collected, and the situation of the selection for testing markers, testing strategy and the testing method and other relevant aspects were sorted out and analyzed by charts.Results:The selection of the testing markers was consistent, but HTLV testing item was added in some member laboratories. The detection strategy of using two ELISA reagents and one nucleic acid testing (NAT) reagent simultaneously was adopted in 47 member blood stations; 3) NAT method was dominated by mini pool-NAT in member laboratories. The number of members adopting mini-pools of 8 (MP8)-NAT decreased from 17 in 2017 to 14 in 2021, while the number of members adopting mini-pools of 6 (MP6)-NAT increased from 13 in 2017 to 22 in 2021; Roche NAT system accounted for the largest proportion.Conclusions:In order to ensure blood safety and avoid missing detection, the blood stations still adopt the detection strategy of using two ELISA reagents and one nucleic acid testing (NAT) reagent simultaneously; Meanwhile, in order to increase the NAT positive rate, the proportion of mini pool-NAT mainly decreased year by year despite its dominating role, while the proportion of individual donation-NAT increased year by year; NAT method is transiting from mini-pools of 8 (MP8) to mini-pools of 6 (MP6); The proportion of imported NAT system used in NAT laboratory is relatively large.

Aim To observe the effect of Gupi Xiaoji Decoction (GPXJY) on the structure and function of mitochondria of human hepatoma cell HepG2 cells and explore its possible mechanism. Methods CCK8 was used to detect cell proliferation, Mito-Tracker Green fluorescence staining was used to observe the mitochondrial structure, flow cytometry was used to detect the membrane potential, Elisa was used to detect the ATP content, fluoroscopic electron microscopy was used to observe the microstructure changes, and high-content screening(HCS) was used to detect the related proteins. Results Fluorescence staining showed that GPXJY damaged the mitochondria of HepG2 cells and decreased the content of ATP. The results of flow cytometry showed that GPXJY could reduce the mitochondrial membrane potential of HepG2 cells. The results of electron microscope showed that GPXJY made the mitochondria of cancer cells swell and so on. HCS found that GPXJY significantly reduced the average fluorescence intensity of Bcl-2 in HepG2 cells, and significantly increased the average fluorescence intensity of apoptosis promoting proteins Bax, cytochrome-c, caspase-3 and cleaved-caspase-3, which was statistically significant. Conclusion GPXJY can regulate the structure and function of mitochondria in HepG2 cells.

Anthocyanidin reductase (ANR) is one of the key enzyme in the flavonoid biosynthetic pathway, and its catalytic activity is important for the synthesis of plant anthocyanin. In this study, specific primers were designed according to the transcriptome data of Lonicera japonica Thunb., and the CDS, gDNA and promoter sequences of ANR genes from Lonicera japonica Thunb. and Lonicera japonica Thunb. var. chinensis (Wats.) Bak. were cloned. The results showed that the CDS sequences of LjANR and rLjANR were 1 002 bp, the gDNA sequences were 2 017 and 2 026 bp respectively, and the promoter sequences were 1 170 and 1 164 bp respectively. LjANR and rLjANR both contain 6 exons and 5 introns, which have the same length of exons and large differences in introns. The promoter sequences both contain a large number of light response, hormone response and abiotic stress response elements. Bioinformatics analysis showed that both LjANR and rLjANR encoded 333 amino acids and were predicted to be stable hydrophobic proteins without transmembrane segments and signal peptides. The secondary structures of LjANR and rLjANR were predicted to be mainly consisted of α-helix and random coil. Sequence alignment and phylogenetic analysis showed that LjANR and rLjANR had high homology with Actinidia chinensis var. chinensis, Camellia sinensis and Camellia oleifera, and were closely related to them. The expression levels of LjANR and rLjANR were the highest in flower buds and the lowest in roots. The expression patterns at different flowering stages were similar, with higher expression levels in S1 and S2 stages and then gradually decreased until reaching the lowest level in S4 stage, after a slow increase in S5 stage, the expression levels decreased again. The expression levels of ANR genes in the two varieties showed significant differences in roots, S2 and S5 stages, while the differences in stems, flower buds, S1, S3 and S6 stages were extremely significant. The prokaryotic expression vector pET-32a-LjANR was constructed for protein expression. The target protein was successfully expressed of about 59 kD. This study lays a foundation for further study on the function of ANR gene and provides theoretical guidance for breeding new varieties of Lonicera japonica Thunb.

Functional reference limits describe key changes in the physiological relationship between a pair of physiologically related components. Statistically, this can be represented by a significant change in the curvature of a mathematical function or curve (e.g., an observed plateau). The point at which the statistical relationship changes significantly is the point of curvature inflection and can be mathematically modeled from the relationship between the interrelated biomarkers. Conceptually, they reside between reference intervals, which describe the statistical boundaries of a single biomarker within the reference population, and clinical decision limits that are often linked to the risk of morbidity or mortality and set as thresholds. Functional reference limits provide important physiological and pathophysiological insights that can aid laboratory result interpretation. Laboratory professionals are in a unique position to harness data from laboratory information systems to derive clinically relevant values. Increasing research on and reporting of functional reference limits in the literature will enhance their contribution to laboratory medicine and widen the evidence base used in clinical decision limits, which are currently almost exclusively contributed to by clinical trials. Their inclusion in laboratory reports will enhance the intellectual value of laboratory professionals in clinical care beyond the statistical boundaries of a healthy reference population and pave the way to them being considered in shaping clinical decision limits. This review provides an overview of the concepts related to functional reference limits, clinical examples of their use, and the impetus to include them in laboratory reports.