ObjectiveTo investigate the correlation between iron metabolism parameters and various syndrome types of unstable angina pectoris （UAP）. MethodsA cross-sectional study was conducted from October 2021 to October 2023， encompassing 213 patients diagnosed with UAP at Xiyuan Hospital of Chinese Academy of Chinese Medical Sciences. Additionally， 30 healthy individuals were selected as control cases. Single-factor analysis was used to investigate the differences in clinical data among different Traditional Chinese Medicine （TCM） syndrome types of UAP and their correlation with iron metabolism indices. The study conducted a comparative analysis of the aforementioned clinical data among patients with and without heat-toxic and blood-stasis syndrome. Logistic regression was used to analyze the correlation between TCM syndrome types and related factors. The receiver operator characteristic （ROC） curve was employed to assess the predictive value of iron metabolism indices， along with their sensitivity and specificity. ResultsCompared to those in the control group， serum iron （SI） and serum ferritin （SF） levels were significantly increased in the UAP group （P<0.01）， while transferrin （TRF） and total iron binding capacity （TIBC） levels were decreased （P<0.01）. However， there was no significant difference in unsaturated iron binding capacity （UIBC）. Multivariate binary Logistic regression analysis identified apolipoprotein A1 （ApoA1）， homocysteine （HCY）， high-sensitivity C-reactive protein （hs-CRP）， and SF as independent influencing factors for the UAP patients （P<0.05， P<0.01）. Additionally， statistically significant differences were observed in SI， SF， TRF， and TIBC among 213 patients with different TCM types （P<0.01）. Patients with heat-toxic and blood-stasis syndrome had higher SI and SF values than those without the syndrome （P<0.01）， while their TIBC and TRF values were lower （P<0.01）. Multivariate binary logistic regression analysis showed that SI and LDL-C levels were closely associated with the differentiation of heat-toxic and blood-stasis syndrome. ConclusionUAP patients often experience iron metabolism disorders， and the heat-toxic and blood-stasis syndrome are significantly correlated with iron metabolism parameters. The SI and LDL-C levels have high specificity and sensitivity in diagnosing heat-toxic and blood-stasis syndrome.

Probiotics are natural systems bridging synthetic biology, physical biotechnology, and immunology, initiating innate and adaptive anti-tumor immune activity. We previously constructed an all-in-one engineered food-grade probiotic Lactococcus lactis (FOLactis) which could boost the crosstalk among different immune cells such as dendritic cells (DCs), natural killer cells, and T cells. Herein, considering the limited clinical efficacy of naked personalized neoantigen peptide vaccines, we decorate FOLactis with tumor antigens by employing a Plug-and-Display system comprising membrane-inserted peptides. Intranodal injection of FOLactis coated with neoantigen peptides (Ag-FOLactis) induces robust DCs presentation and neoantigen-specific cellular immunity. Notably, Ag-FOLactis not only triggers a 45-fold rise in the quantity of locally reactive neoantigen-specific T cells but also induces epitope spreading in both subcutaneous and metastatic tumor-bearing models, leading to potent inhibition of tumor growth. These findings imply that Ag-FOLactis represents a powerful platform to rapidly and easily display antigens, facilitating the development of a bio-activated platform for personalized therapy.

[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×10⁸vs (6.49±2.16)×10⁸, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.

OBJECTIVE: To carry out pathological and genetic analyses on a fetus with intrauterine growth restriction and death during second trimester after induced abortion. METHODS: A fetus undergone induced abortion due to intrauterine growth restriction and death during second trimester at the the Seventh Affiliated Hospital of Sun Yat-Sen University in 2024 was selected as the study subject. Clinical data of the pregnancy were collected. DNA was extracted from tissues from the aborted fetus and peripheral blood samples from its parents. Chromosomal microarray analysis and whole exome sequencing were carried out. Candidate variants were verified by Sanger sequencing. Following abortion, routine autopsy and pathological analysis were conducted. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: KY-2025-334-01). RESULTS: The aborted fetus was a male and harbored compound heterozygous nonsense variants of the TH gene (c.457C>T/p.Arg153* and c.694C>T/p.Gln232*), for which both parents were heterozygous carriers. Autopsy and pathological analysis revealed that the fetus had pathological features including loose arrangement of myocardial fibers and congestion in the liver. CONCLUSION Biallelic null variants of the TH gene may cause heart failure by affecting the development of cardiovascular system, which in turn may lead to intrauterine death. This study has provided new clues for the molecular diagnosis of stillbirth and recurrent pregnancy loss.
Humans ; Female ; Pregnancy ; Male ; Heterozygote ; Codon, Nonsense/genetics* ; Fetal Growth Retardation/pathology* ; Adult ; Stillbirth/genetics*

Objective:To develop and assess a three-class machine learning model for predicting wild-type, 19 del, and 21 L858R mutations of the epidermal growth factor receptor (EGFR) in lung adenocarcinoma using 18F-FDG PET/CT radiomic features and clinical features. Methods:The retrospective data was collected from 703 patients (346 males, 357 females; age (64.3±9.0) years) with lung adenocarcinoma at the First People′s Hospital of Changzhou from January 2018 to June 2023. Patients were divided into the training set (563 cases) and test set (140 cases) at the ratio of 8∶2. Clinical features were selected using recursive feature elimination (RFE). Radiomic features were extracted from PET and CT images, and the optimal feature sets were selected using minimum redundancy maximum relevance (mRMR) and least absolute shrinkage and selection operator (LASSO) methods. Base models were constructed by using random forest (RF), logistic regression (LR), support vector machine (SVM), K-nearest neighbors (KNN), and multi-layer perceptron (MLP), and the stacking method was applied to establish the CT and PET ensemble models. Delong test was used to compare the AUC differences between the PET/CT combined model and the clinical + PET/CT integrated model.Results:Among 703 patients, 273 were with EGFR wild-type, 202 were with 19 del mutation, and 228 were with 21 L858R mutation. In the single-modal analysis, the AUCs of CT ensemble model in the training and test sets were 0.893 and 0.667, respectively, while the AUCs of PET ensemble model were 0.692 and 0.660. The AUC of PET/CT combined model were 0.897 in training set and 0.672 in test set. The AUC of clinical + PET/CT integrated model showed further improvement, with AUCs of 0.902 and 0.721 in training and test sets, respectively. Notably, the clinical + PET/CT integrated model outperformed PET/CT combined model in predicting wild-type EGFR (test set AUC: 0.784 vs 0.707; Z=3.28, P=0.001). Conclusion:The three-class model (clinical + PET/CT integrated model) based on 18F-FDG PET/CT radiomics and clinical features effectively predicts EGFR mutation subtypes in lung adenocarcinoma.

Objective To conduct a unique and pioneering molecular epidemiological investigation of a case of Eurasian avian-like H1N1 swine influenza identified in Yunnan Province in 2022(the fourth such case in the province)and to understand its genetic characteristics so as to reveal its potential impact on human health.Methods Real-time fluorescent quantitative PCR detection technology was used for the nucleic acid testing of the case's pharyngeal swab samples,close contacts,and environmental samples from the living area.Positive samples were subjected to virus isolation using MDCK cells.Cell cultures were authenticated using erythrocyte agglutination assay with guinea pig blood and real-time fluorescence quantitative RT-PCR.Whole genome sequencing was performed using the Illumina MiseqNext-generation sequencing platform,and a phylogenetic tree was constructed using MEGA 7.0 software to analyze the genetic molecular characteristics.Results The first G5 genotype Eurasian avian-like H1N1 swine influenza virus in Yunnan Province was successfully isolated,and the whole genome sequence of the virus was obtained.This virus possessed the molecular characteristics associated with increased adaptability,virulence,or transmissibility in mammals and had a nucleotide consistency of 99.2％～99.7％with a porcine strain isolated in Jiangsu province.These findings underscored the potential threat this virus poses to human health.Conclusion The study underscores the importance of further monitoring swine influenza in preventing new influenza virus subtypes that can infect humans.

The incidence of less common tumors is intermediate between rare tumors and high-prevalence tumors,while these less common tumors such as biliary tract cancers generate a significant regional health burden. The overall incidence of less common tumors is relatively low, and thus their clinical epidemiological studies face challenges such as recruitment difficulties,poor representation,and low standardization. Surgical center-based disease-specific cohorts have the advantages of case concentration,complete samples,and well-developed data,which are uniquely valuable in clinical epidemiological studies. Taking the disease-specific cohort of biliary tract cancers as an example,the authors combed through the relevant references and summarized the thinking in the practice of constructing disease-specific cohorts of less common tumors based on surgical centers. The architecture of the disease-specific cohort construction has been generalized as follows: the hardware includes a database, a biobank, and a platform of information synchronization, and the software follows the design principle of “high cohesion and low coupling”. The authors also recommend an orderly expansion of study size and implementation of quality control through all segments of cohort construction, and hope that these reflections could provide a reference for similar disease-specific cohorts.

Objective To examine the effect and mechanism of Duhuo Jisheng Decoction on lumbar intervertebral disc degeneration.Methods In total,105 Sprague-Dawley(SD)rats were randomly assigned to seven groups:sham operation group,model group,Duhuo Jisheng Decoction 1,rapamycin group,rapamycin+Duhuo Jisheng Decoction group,MHY1485 group,and MHY1485+Duhuo Jisheng Decoction group.Except for the sham operation group,the other groups underwent annulus fibrosus puncture to establish a lumbar intervertebral disc degeneration(IDD)animal model.After successful model establishment,a six-week drug intervention was performed,followed by MRI evaluation of the degree of disc degeneration.Samples of the L5-6 intervertebral disc were collected for HE and Alcian blue-nuclear fast red staining,and the degeneration of the nucleus pulposus and changes in the extracellular matrix were observed using light microscopy.Simultaneously,the expression levels of the downstream proteins of the mTOR pathway,p70S6K and 4E-BP1,along with the autophagy-related genes Beclin-1 and LC3,were assessed at both the protein and mRNA levels.Autolysosomes in nucleus pulposus cells were visualized using transmission electron microscopy(TEM).Results ①MRI showed disc Pfirrmann grade I in the sham group,and disc Pfirrmann grade Ⅲ-Ⅳ in the remaining 6 groups in L5-6 segments.②The degree of lumbar disc degeneration with histomorphological observation:MHY1485 group>model group>MHY1485 group+Duhuo Jisheng Decoction group>rapamycin group>Duhuo Jisheng Decoction group>rapamycin+Duhuo Jisheng Decoction group>sham group.Allan-nuclear red staining extracellular matrix proteoglycan content:sham group>rapamycin+Duhuo Jisheng Decoction group>rapamycin group>MHY1485 group+Duhuo Jisheng Decoction group>model group>MHY1485 group.③Relative expression of p-mTOR,p70S6K and 4E-BP1 downstream of mTOR signaling pathway:MHY1485 group>MHY1485 group+Duhuo Jisheng Decoction group>model group>rapamycin group>Duhuo Jisheng Decoction group>rapamycin+Duhuo Jisheng Decoction group>sham group,each experimental group varied significantly from the model group(P<0.01).④Autophagy-related protein Beclin-1 and LC3 expression levels:rapamycin+Duhuo Jisheng Decoction group>Duhuo Jisheng Decoction group>rapamycin>MHY1485 group+Duhuo Jisheng Decoction group>model group>MHY1485>sham group,and the content of each group was significantly(P<0.01).⑤TEM observation of autophagy levels in the cells of each group showed the formation of autolysosomes in Duhuo Jisheng Decoction group,rapamycin group andrapamycin+Duhuo Jisheng Decoction group.Conclusion Duhuo Jisheng Decoction can slow down the degenerative process of lumbar intervertebral discs in rats,and its effect may be linked to the suppression of the mTOR signaling pathway and the promotion of autophagy in nucleus pulposus cells.

BACKGROUND:Spinal osteosarcoma is a rare and highly aggressive malignant tumor.Most existing studies are based on small sample sizes and have inconsistent results,making it difficult to provide reliable clinical guidance.Especially in China,due to the low incidence of spinal osteosarcoma and limited related research,clinicians lack effective prognostic tools during treatment.OBJECTIVE:To construct and validate a nomogram model for predicting the survival of spinal osteosarcoma patients based on the Surveillance,Epidemiology,and End Results(SEER)database,providing scientific evidence for clinical decision-making,particularly for optimizing treatment plans for Chinese patients.METHODS:This study conducted a retrospective analysis of patient data diagnosed with spinal osteosarcoma from the SEER database between 2000 and 2021.First,independent prognostic factors associated with specific mortality from spinal osteosarcoma were identified through univariate and multivariate Cox proportional hazards models.Subsequently,these independent prognostic factors were used to construct a nomogram model for predicting survival rates of spinal osteosarcoma patients using the"rms"package in RStudio.The model's discrimination was assessed using the C-index.Predictive ability was validated through receiver operating characteristic curves and area under the curve values.Calibration was evaluated by calibration plots,and clinical value was measured using decision curve analysis.Additionally,Kaplan-Meier survival analysis was performed to assess the rationality of the nomogram groupings.RESULTS AND CONCLUSION:(1)The final model included six variables:chemotherapy,tumor size,histological type,grade,race,and surgical intervention.(2)The C-indices of the model in the training and validation sets were 0.685 and 0.673,respectively,indicating good discrimination.(3)Calibration curves showed high consistency between predicted survival probabilities and actual survival probabilities.(4)Decision curve analysis indicated that the model provided significant net benefits across a wide range of mortality risks.(5)Kaplan-Meier survival analysis revealed significant differences in prognosis between high-risk and low-risk groups.(6)The constructed nomogram model accurately predicts the 1-year,2-year,and 3-year survival rates of spinal osteosarcoma patients,demonstrating high clinical applicability.This model not only provides an effective survival prediction tool for American patients but also offers important insights for optimizing treatment plans for spinal osteosarcoma patients in China.Future research should further validate the model's applicability in different populations and explore the impact of novel treatment methods on the prognosis of spinal osteosarcoma,aiming to improve the survival rates and quality of life of patients in China.

Objective To construct a clinical-radiological nomo-gram model for severe mycoplasma pneumoniae pneumonia coinfec-tion with other pathogens(Co-SMPP)in children.Methods The clinical and radiological data of children with severe mycoplasma pneumoniae pneumonia(SMPP)who underwent nucleic acid testing or bronchoalveolar lavage(BAL)were analyzed retrospectively.The data analysis were performed by using SPSS 27.0 software.The group comparison between simple SMPP and Co-SMPP children was conducted by using t-tests,Mann-Whitney U tests,or chi-square tests.Nomogram analysis was performed by using R software and rms packages.The predictive performance of the model was evaluated by using the receiver operating characteristic(ROC)curve.Results A total of 194 SMPP children were included in the study,including 136 cases(70.1％)with simple SMPP,58 cases(29.9％)with Co-SMPP.The fibrinogen and albumin levels were lower in Co-SMPP children[(3.53±0.85)g/L,41.00(39.03,43.68)g/L]than in simple SMPP children[(3.79±0.80)g/L,42.80(41.00,44.40)g/L],with P values of 0.047 and 0.036,respec-tively.The probability of bronchial stenosis and grid shadow were higher in Co-SMPP children than in simple SMPP children,and there were significant differences between the two groups(P＜0.001,P=0.010).The odds ratio of bronchial stenosis in predicting Co-SMPP children was 14.085.The clinical-radiological nomogram model had an area under the curve(AUC)of 0.840,with sensi-tivity and specificity of 0.756 and 0.848,respectively.Conclusion The nomogram model based on clinical-radiological features can effectively predict Co-SMPP.