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 clinical outcomes of sublingual gland flap in the repair of postoperative jaw defects because of chronic phosphorous osteomyelitis(PNJ)and medication-related osteonecrosis of the jaw(MRONT).Methods:3 patients with PNJ and 2 with MRONJ were treated by sublingual gland flap in the repair of the postoperative jaw defet.The treatment effects were evalua-ted by clinical obseration.Results:In the 5 patients the wound healed well within 2 weeks postoperatively,with no infection and with normal sublingual gland function after surgery.Conclusion:The use of sublingual gland flap is an effective and feasible method for re-pairing local defects in the mandibles after surgery for chronic PNJ and MRONJ.

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.

Objective To understand current epidemic trend of coronavirus disease-2019(COVID-19)in Anhui province in the optimization policy stage,and to analyze the pathogenic characteristics of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)in different population.Methods Using a cross-sectional survey design,from December 19 to 20,2022,an online questionnaire survey was conducted among residents of Anhui province through the official Wechat public accounts of provincial and municipal institutions with high traffic,to collect infor-mation on the incidence and clinic situation of COVID-19.The chi-square test was used to compare the proportion of COVID-19 suspected symptoms in different regions,ages and occupations.Results A total of 69 014 question-naires were distributed and 68 232 valid questionnaires were recovered with an effective rate of 98.97％.The pro-portion of the participants with COVID-19 suspected symptoms in the past 2 weeks was 51.37％,of which 77.88％self-medicated at home.The top three cities were Bozhou,Fuyang and Bengbu.The age group of 15-59 had the highest proportion of COVID-19 suspected symptoms(51.96％).Among various occupations,service providers had the highest proportion of COVID-19 suspected symptoms(61.07％).70.20％of the respondents felt anxious about the infection of SARS-CoV-2 and thought it was more serious than the flu.Conclusion The relatively high number of the infected cases and the anxiety of the people are all challenges faced by Anhui province in the stage of optimizing policies.Under the new situation of the epidemic,it is necessary to continuously monitor the local preva-lent strains and strengthen the monitoring of clinical symptoms of the infected cases,and effectively control the speed of the virus spread through public health policies and various economic and publicity measures,so as not to cause a run on medical resources and excessive excess deaths.

177Lu-prostate specific membrane antigen(PSMA)radio-ligand therapy has been approved abroad for advanced prostate cancer and has been in several clinical trials in China.Based on domestic clinical practice and experimental data and referred to international experience and viewpoints,the expert group forms a consensus on the clinical application of 177Lu-PSMA radio-ligand therapy in prostate cancer to guide clinical practice.

Objective To prepare a nano drug(PFOB@Lip-MMC)with liposome as the carrier,liquid perfluorooc-tyl bromide(PFOB)as core and mitomycin C(MMC)loading on the liposome shell and study its inhibitory effect on the proliferation of human pterygium fibroblasts(HPFs).Methods The thin film dispersion-hydration ultrasonic method was used to prepare PFOB@Lip-MMC and detect its physical and chemical properties.Cell Counting Kit-8,Cam-PI cell viability staining and flow cytometry were employed to detect the impact of different concentrations of PFOB@Lip-MMC on the via-bility of HPFs.DiI fluorescence labeled PFOB@Lip-MMC was used to observe the permeability of the nano drug to HPFs under a laser confocal microscope.After establishing HPF inflammatory cell models,they were divided into the control group(with sterile phosphate-buffered saline solution added),PFOB@Lip group(with PFOB@Lip added),MMC group(with MMC added),PFOB@Lip-MMC group(with PFOB@Lip-MMC added)and normal group(with fresh culture medi-um added)according to the experimental requirements.After co-incubation for 24 h,flow cytometer was used to detect the apoptosis rate of inflammatory cells,and the gene expression levels of interleukin(IL)-1β,prostaglandin E2(PGE2),tumor necrosis factor(TNF)-α and vascular endothelial growth factor(VEGF)in cells were analyzed by PCR.Results The average particle size and Zeta potential of PFOB@Lip-MMC were(103.45±2.17)nm and(27.34±1.03)mV,respec-tively,and its entrapped efficiency and drug loading rate were(72.85±3.28)％and(34.27±2.04)％,respectively.The sustained-release MMC of drug-loaded nanospheres reached(78.34±2.92)％in vitro in a 24-hour ocular surface environ-ment.The biological safety of PFOB@Lip-MMC significantly improved compared to MMC.In terms of the DiI fluorescence labeled PFOB@Lip-MMC,after co-incubation with inflammatory HPFs for 2 h,DiI fluorescence labeling was diffusely dis-tributed in the cytoplasm of inflammatory HPFs.The apoptosis rate of inflammatory HPFs in the PFOB@Lip-MMC group[(77.23±4.93)％]was significantly higher than that in the MMC group[(51.62±3.28)％].The PCR examination results showed that the gene transcription levels of IL-1 β,PGE2,TNF-α and VEGF in other groups were significantly reduced com-pared to the control group and PFOB@Lip group,with the most significant decrease in the PFOB@Lip-MMC group(all P＜0.05).Conclusion In this study,a novel nano drug(PFOB@LIP-MMC)that inhibited the proliferation of HPFs was successfully synthesized,and its cytotoxicity was significantly reduced compared to the original drugs.It has good bio-compatibility and anti-inflammatory effects,providing a new treatment approach for reducing the recurrence rate after pte-rygium surgery.

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.