Based on whole-genome identification of the TPS gene family in Perilla frutescens and screening, cloning, bioinformatics, and expression analysis of the synthetic enzyme for the insect-resistant component germacrene D, this study lays the foundation for understanding the biological function of the TPS gene family and the insect resistance mechanism in P. frutescens. This study used bioinformatics tools to identify the TPS gene family of P. frutescens based on its whole genome and predicted the physicochemical properties, systematic classification, and promoter cis-elements of the proteins. The relative content of germacrene D was detected in both normal and insect-infested leaves of P. frutescens, and the germacrene D synthase was screened and isolated. Gene cloning, bioinformatics analysis, and expression profiling were then performed. The results showed that a total of 99 TPS genes were identified in the genome, which were classified into the TPS-a, TPS-b, TPS-c, TPS-e/f, and TPS-g subfamilies. Conserved motif analysis showed that the TPS in P. frutescens has conserved structural characteristics within the same subfamily. Promoter cis-element analysis predicted the presence of light-responsive elements, multiple hormone-responsive elements, and stress-responsive elements in the TPS family of P. frutescens. Transcriptome data revealed that most of the TPS genes in P. frutescens were highly expressed in the leaves. GC-MS analysis showed that the relative content of germacrene D significantly increased in insect-damaged leaves, suggesting that it may act as an insect-resistant component. The germacrene D synthase gene was screened through homologous protein binding gene expression and was found to belong to the TPS-a subfamily, encoding a 64.89 kDa protein. This protein was hydrophilic, lacked a transmembrane structure and signal peptide, and was predominantly expressed in leaves, with significantly higher expression in insect-damaged leaves compared to normal leaves. In vitro expression results showed that germacrene D synthase tended to form inclusion bodies. Molecular docking showed that farnesyl pyrophosphate(FPP) fell into the active pocket of the protein and interacted strongly with six active sites. This study provides a foundation for further research on the biological functions of the TPS gene family in P. frutescens and the molecular mechanisms underlying its insect resistance.
Perilla frutescens/chemistry* ; Plant Proteins/chemistry* ; Multigene Family ; Sesquiterpenes, Germacrane/metabolism* ; Alkyl and Aryl Transferases/chemistry* ; Phylogeny ; Gene Expression Regulation, Plant

OBJECTIVE: To explore, identify, and develop novel blood-based indicators using machine learning algorithms for accurate preoperative assessment and effective prediction of postoperative complication risks in patients with rheumatoid arthritis (RA) undergoing total knee arthroplasty (TKA). METHODS: A retrospective cohort study was conducted including RA patients who underwent unilateral TKA between January 2019 and December 2024. Inpatient and 30-day postoperative outpatient follow-up data were collected. Six machine learning algorithms, including decision tree, random forest, logistic regression, support vector machine, extreme gradient boosting, and light gradient boosting machine, were used to construct predictive models. Model performance was evaluated using the area under the receiver operating characteristic curve (AUC), F1-score, accuracy, precision, and recall. SHapley Additive exPlanations (SHAP) values were employed to interpret and rank the importance of individual variables. RESULTS: According to the inclusion criteria, a total of 1 548 patients were enrolled. Ultimately, 18 preoperative indicators were identified as effective predictive features, and 8 postoperative complications were defined as prediction labels for inclusion in the study. Within 30 days after surgery, 453 patients (29.2%) developed one or more complications. Considering overall accuracy, precision, recall, and F1-score, the random forest model [AUC=0.930, 95% CI (0.910, 0.950)] and the extreme gradient boosting model [AUC=0.909, 95% CI (0.880, 0.938)] demonstrated the best predictive performance. SHAP analysis revealed that anti-cyclic citrullinated peptide antibody, C-reactive protein, rheumatoid factor, interleukin-6, body mass index, age, and smoking status made significant contributions to the overall prediction of postoperative complications. CONCLUSION Machine learning-based models enable accurate prediction of postoperative complication risks among RA patients undergoing TKA. Inflammatory and immune-related blood biomarkers, such as anti-cyclic citrullinated peptide antibody, C-reactive protein, and rheumatoid factor, interleukin-6, play key predictive roles, highlighting their potential value in perioperative risk stratification and individualized management.
Humans ; Arthroplasty, Replacement, Knee/adverse effects* ; Arthritis, Rheumatoid/blood* ; Machine Learning ; Postoperative Complications/blood* ; Female ; Male ; Retrospective Studies ; Middle Aged ; Aged ; Risk Factors ; Preoperative Period ; C-Reactive Protein/analysis* ; Risk Assessment

Objective To investigate the diagnostic value of quantitative energy spectrum CT and multi-parameter magnetic resonance imaging(MRI)on epidermal growth factor receptor(EGFR)mutation in lung adenocarcinoma.Methods Clinical data of 93 patients diagnosed with lung adenocarcinoma and EGFR mutation status by pathology in Jingdezhen Hospital of Traditional Chinese Medicine from June 2020 to June 2022 were retrospectively analyzed.They were divided into mutation group(52 cases)and wild-type group(41 cases)according to EGFR mutation status,and all patients were given quantitative spectral CT and MRI examinations.Changes in parameters between two groups were compared.Results CT40kev,CT70kev,spectral curve slope(λHU),effective atomic number and iodine concentration in arterial and venous phases in mutation group were significantly higher than those in wild-type group(P<0.05).Apparent diffusion coefficient(ADC)and true diffusion coefficient(D)of mutant group were significantly higher than those of wild-type group,while volume transport constant(Ktrans)and reflux rate constant(Kep)of mutant group were significantly lower than those of wild-type group(P<0.05).Analysis of receiver operating characteristics curve showed that area under the curve(AUC)value for CT40kev,CT70kev,λHU,effective atomic number,iodine concentration,CT comprehensive parameters,ADC,D,Ktrans,Kep and MRI comprehensive parameters for diagnosing EGFR mutation status in lung adenocarcinoma were 0.615,0.622,0.680,0.715,0.655,0.855,0.899,0.795,0.740,0.751 and 0.968 respectively.There was statistically significant difference in AUC between CT comprehensive parameters and MRI comprehensive parameters(P<0.05).Conclusion Both quantitative energy spectrum CT and multi-parameter MRI have high diagnostic value in the diagnosis of EGFR mutation status of lung adenocarcinoma,but MRI has higher diagnostic value,which is helpful to provide objective basis for accurate assessment of EGFR mutation status.

AIM:To investigate the effect of Cara-gana sinica root(CSR)on ferroptosis in the Erastin-induced chondrocyte ferroptosis model and possi-ble mechanisms.METHODS:The C28/I2 chondro-cyte cell line was cultured to construct a cell model of Erastin-induced ferroptosis.Cell viability was de-tected by MTT assay;cell death was observed by Calcein/PI staining;cell lactate dehydrogenase(LDH)and total glutathione(GSH)levels were de-tected by the kit;reactive oxygen species(ROS)lev-els were detected by fluorescent probe BODIPY 581/591 C11 labeling;mitochondrial membrane po-tential(ΔΨm)changes were observed by Rh123 and JC-1 staining;Western blot was used to detect the expression of ferroptosis-related proteins(AC-SL4,GPX4)and TRPM7 proteins.RESULTS:Erastin treatment decreased chondrocyte viability,in-creased cytotoxicity,induced oxidative stress,dis-rupted ΔΨm,and up-regulated ACSL4 protein ex-pression,while down-regulating GPX4 protein ex-pression and inducing chondrocyte ferroptosis.In contrast,CSR restored cell viability and reduced oxi-dative stress,thereby inhibiting chondrocyte fer-roptosis.In addition,CSR reduced the Erastin-in-duced increase in TRPM7 protein expression level.CONCLUSION:Erastin induced lipid peroxidation in C28/I2 chondrocytes,causing mitochondrial dam-age and ferroptosis;CSR may inhibit chondrocyte ferroptosis by blocking TRPM7,thus exerting a pro-tective effect on chondrocytes.

Objective:To investigate the clinical characteristics and treatment of patients with CD4+CD8-T-cell large granular lymphocytic leukemia(T-LGLL).Methods:The clinical manifestations,diagnosis and treatment of 1 case of CD4+CD8-T-LGLL patient were reported,and relevant literatures were reviewed.Results:The patient was a 70-year-old woman with slow clinical progress,mainly manifested by thrombocytopenia and myelodysplasia.The blood smear was mainly composed of large granular lymphocytes.Immunotyping and T-cell receptor gene rearrangement analysis showed that it was in line with T-LGLL.Partial remission(PR)was achieved through the treatment of cyclophosphamide(50 mg/d)combined with prednisone(gradually reduced and stopped later).Conclusion:CD4+CD8-T-LGLL is very rare in clinical practice,and its clinical manifestations are different from those of CD4-CD8+T-LGLL.

Objective:To investigate the intention to stay on among primary health care workers (PHWs) during the COVID-19 epidemics and its influencing factors.Methods:An online questionnaire survey was conducted among PHWs from 62 primary health institutions in 31 provinces, autonomous regions and municipalities across China selected by multi-stage whole cluster random sampling method between May and October 2022. According to the job position, the PHWs were divided into 5 categories: general practitioners (GPs), nurses, public health doctors, managers and support staff. Intention to stay was measured using the Chinese version of the Intention to Stay Questionnaire. Multiple linear regression model was used to analyze the influencing factors of the intention to stay in PHWs of different occupational categories, including personal factors, work factors, factors related to the COVID-19 and psychological reactions.Results:A total of 3 769 PHWs from 44 community health service centers, 18 township hospitals of 27 provinces/autonomous regions and 4 municipalities participated in this survey. The mean age of participants was (37.4±9.2) years, including 2 971(78.8%) women. The mean score of intention to stay of participants was 21.7±4.1. Compared with GPs, managers had lower intention to stay ( P=0.004). Age, female, in marriage, monthly income, years of primary care service, self-evaluation of unit support function, MSQ-SF score, and PA score were positively correlated with the score of intention to stay (all P<0.05), while PHQ-9 score, EE score, and DP score were negatively correlated with the score of intention to stay (all P<0.05). The MSQ-SF scores of all occupational categories were positively correlated with the scores of intention to stay (all P<0.01). In addition, among GPs, monthly income, years of service in primary care, self-evaluation of unit support function, and PA scores were positively correlated with the score of intention to stay (all P<0.05), while PHQ-9 scores and EE scores were negatively correlated with the score of intention to stay (all P<0.05). Among nurses, age, female gender, monthly income, self-evaluation of unit support function, and PA score were positively correlated with the score of intention to stay (all P<0.05), while EE scores and retention will score were negatively associated with ( P<0.001). Among public health doctors, in marriage was positively correlated with the score of intention to stay ( P=0.018). Among managers, DP score was negatively correlated with the score of intention to stay ( P=0.001). Among support staff, female gender, monthly income, years of primary care service, self-evaluation of unit support function, and PA scores were positively correlated with the score of intention to stay (all P<0.05), while EE score and DP score were negatively correlated with intention to stay (all P<0.05). Conclusions:The intention to stay of PHWs in China during the COVID-19 was at an intermediate level, which was affected by many factors. Among them, job satisfaction was positively correlated with the intention to stay of all occupational categories, and the influencing factors of different occupational categories have some variations.

OBJECTIVE To explore whether inhaling Bingchangsan(BCS can modulate the Nrf2/HO-1 signaling pathway to improve ferroptosis and cognitive dysfunction in Alzheimer's Disease(AD mouse models.METHODS 30 APP/PS1 mice were ran-domly divided into three groups:a model group(APP/PS1 group,a low-dose Bingchangsan group(BCS-L group,and a high-dose Bingchangsan group(BCS-H group.10 age-matched wild-type(WT mice were used as a control group.The WT group and the APP/PS1 group were treated with nebulized pure water,while the BCS-L group received 0.5μL of Bingchangsan inhalation solution per day,and the BCS-H group received 1μL,both administered via nebulization for 10 min daily for 2 weeks.2 weeks post-treat-ment,spatial cognitive abilities of the mice were assessed using the Morris water maze test.Following the water maze experiment,the mice were euthanized,and their brain tissues were collected for Aβ1-42 immunofluorescence,P-tau immunohistochemistry,and Nissl staining.Hippocampal tissues were extracted for Western blot and qPCR analysis to measure the protein and mRNA expression levels of Keap1,Nrf2,HO-1,GPX4,SCL7A11,TFRC,DMT1,FTL,and FTH1.Additionally,brain tissues were used for glutathione(GSH content measurement using a GSH assay kit.RESULTS In the BCS groups,the latency period for escape shortened,and the time spent in the quadrant with the platform,as well as the number of times crossing the platform,increased.Compared to the APP/PS1 group,the expression levels of Aβ1-42 and P-tau in the hippocampal CA1 region and cortex of mice in the BCS groups were signifi-cantly reduced(P<0.05,P<0.01.Nissl staining revealed a denser arrangement of neurons with more Nissl bodies in the BCS groups.In the hippocampal tissue,compared to untreated APP/PS1 mice,significant increases in the protein and mRNA levels of Nrf2,HO-1,SLC7A11,GPX4,and FTL were observed in all BCS groups(P<0.05,P<0.01.While the BCS-L group showed in-creased levels of FTH1 protein and mRNA,these changes were not statistically significant.Furthermore,the expression levels of TFRC,DMT1,and Keap1 were significantly downregulated in the BCS groups(P<0.05,P<0.01.Additionally,treatment with BCS effectively restored the glutathione(GSH content in the brain(P<0.01.CONCLUSION Inhalation of BCS can improve cognitive dysfunction in APP/PS1 mice.BCS activates the Nrf2/HO-1 pathway,increases the expression of GPX4,and enhances Nrf2 tran-scriptional activity by inhibiting Keap1.This leads to an upregulation of SLC7A11 and restoration of GSH levels,effectively countering ferroptosis.Additionally,BCS effectively inhibits TFRC and DMT1,while upregulating FTL and FTH1 expression,thereby maintaining intracellular iron homeostasis.This contributes to mitigating the impact of ferroptosis on APP/PS1 mice,subsequently enhancing their cognitive functions.

OBJECTIVE To explore the effect of Xuanbai Chengqi Decoction on respiratory and oxygenation functions and the ex-pression levels of serum aquaporin(AQP)1 and AQP5 in patients with severe pneumonia complicated with gastrointestinal dysfunction of lung heat and fu-organ repletion type.METHODS 60 patients with severe pneumonia complicated with gastrointestinal dysfunc-tion of lung heat and fu-organ repletion type were randomly divided into control group and treatment group,with 30 cases each.The control group received standardized Western medicine treatment,and the treatment group was treated with Xuanbai Chengqi Decoction in addition to the control group.Both groups were treated for 7 d.The respiratory rate and oxygenation index,mechanical ventilation u-tilization rate,the clinical score including CURB-65 and CPIS scores,TCM syndrome score,gastrointestinal function indicators inclu-ding intra-abdominal pressure,serum gastrin(GAS)and vasoactive intestinal peptide(VIP),AQP1 and AQP5 levels were compared between the two groups before and after treatment.Ventilation utilization and ICU hospitalization days during treatment were compared between the two groups.RESULTS After treatment,compared with the control group,the respiratory rate,TCM syndrome score and intra-abdominal pressure in the treatment group were decreased significantly(P<0.05,P<0.01);meanwhile,the oxygenation index and the levels of serum GAS,AQP1 and AQP5 were increased significantly(P<0.05,P<0.01).CONCLUSION Xuanbai Chengqi Decoction can significantly improve clinical symptoms such as respiratory and oxygenation functions in patients,and its mechanism may be related to the regulation of AQP1 and AQP5.

convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.