1.Spatiotemporal Electrical Impedance Tomography for Speech Respiratory Assessment in Cleft Palate: an Interpretable Machine Learning Study
Yang WU ; Xiao-Jing ZHANG ; Hao YU ; Cheng-Hui JIANG ; Bo SUN ; Jia-Feng YAO
Progress in Biochemistry and Biophysics 2026;53(2):485-500
ObjectiveCleft palate (CP) is a common congenital deformity often associated with velopharyngeal insufficiency (VPI), which disrupts the physiological coupling between respiration and speech. Conventional clinical assessments, such as nasometry and spirometry, provide limited static data and fail to visualize the dynamic spatiotemporal distribution of lung ventilation during phonation. This study introduces spatiotemporal electrical impedance tomography (ST-EIT) to evaluate speech-respiratory functional features in CP patients compared to normal controls (NC). The aim is to characterize multi-domain respiratory patterns and to validate an interpretable machine learning framework for providing objective, quantitative evidence for clinical assessment. MethodsSeventy-five participants were enrolled in this study, comprising 37 patients with surgically repaired CP and 38 healthy volunteers matched for age, gender, and body mass index (BMI). All subjects performed standardized sustained phonation tasks while undergoing synchronous monitoring with a 16-electrode EIT system and a pneumotachograph. A comprehensive feature engineering pipeline was developed to extract physiological parameters across 3 complementary domains. (1) Temporal domain: including inspiratory/expiratory phase duration (tPhase), time constants (Tau), and inspiratory-to-expiratory time ratios (TI/TE); (2) airflow domain: comprising mean flow, peak flow, and instantaneous flow at 25%, 50%, and 75% of tidal volume; and (3) spatial domain: quantifying global and regional tidal impedance variation (TIV), global inhomogeneity (GI), and center of ventilation (CoV). Extreme Gradient Boosting (XGBoost) classifiers were trained using 5 distinct data sources (Spirometry, Nasometry, Inspiratory-EIT, Expiratory-EIT, and fused ST-EIT). Model performance was rigorously evaluated via stratified 5-fold cross-validation, and Shapley additive explanations (SHAP) were employed to quantify global and local feature contributions. ResultsThe CP group exhibited a distinct respiratory phenotype compared to controls. In the temporal domain, CP patients showed significantly shorter inspiratory (1.60 s vs.1.85 s, P<0.001) and expiratory phase durations (2.45 s vs. 3.95 s, P<0.001), indicating a rapid, shallow breathing rhythm. In the airflow domain, while inspiratory flows were comparable, the CP group demonstrated significantly elevated mean and peak flows during the expiratory phase (P<0.001), reflecting compensatory respiratory effort. Spatially, CP patients presented significant ventilation redistribution, characterized by higher regional TIV in the right-anterior (ROI1) and left-posterior (ROI4) quadrants, but lower TIV in the left-anterior (ROI2) quadrant. In terms of diagnostic accuracy, the multi-modal ST-EIT model achieved the highest performance (AUC: 0.915±0.012, Accuracy: 0.843±0.019, F1-score: 0.872±0.017), substantially outperforming models based on spirometry (AUC: 0.721) or nasometry (AUC: 0.625) alone. Interpretability analysis revealed that spatial domain features were the most critical, contributing 53.4% to the model’s decision-making, followed by temporal (25.0%) and airflow (21.6%) features. ConclusionST-EIT successfully captures the temporal, airflow, and spatial deviations in CP speech respiration that are undetectable by conventional methods—specifically, rapid phase transitions, hyperdynamic expiratory airflow, and regional ventilation heterogeneity. This study validates ST-EIT as a robust, non-invasive, and radiation-free tool for characterizing speech-respiratory dysfunction, offering high clinical value for bedside screening, rehabilitation planning, and longitudinal monitoring of patients with cleft palate.
2.Spatiotemporal Electrical Impedance Tomography for Speech Respiratory Assessment in Cleft Palate: an Interpretable Machine Learning Study
Yang WU ; Xiao-Jing ZHANG ; Hao YU ; Cheng-Hui JIANG ; Bo SUN ; Jia-Feng YAO
Progress in Biochemistry and Biophysics 2026;53(2):485-500
ObjectiveCleft palate (CP) is a common congenital deformity often associated with velopharyngeal insufficiency (VPI), which disrupts the physiological coupling between respiration and speech. Conventional clinical assessments, such as nasometry and spirometry, provide limited static data and fail to visualize the dynamic spatiotemporal distribution of lung ventilation during phonation. This study introduces spatiotemporal electrical impedance tomography (ST-EIT) to evaluate speech-respiratory functional features in CP patients compared to normal controls (NC). The aim is to characterize multi-domain respiratory patterns and to validate an interpretable machine learning framework for providing objective, quantitative evidence for clinical assessment. MethodsSeventy-five participants were enrolled in this study, comprising 37 patients with surgically repaired CP and 38 healthy volunteers matched for age, gender, and body mass index (BMI). All subjects performed standardized sustained phonation tasks while undergoing synchronous monitoring with a 16-electrode EIT system and a pneumotachograph. A comprehensive feature engineering pipeline was developed to extract physiological parameters across 3 complementary domains. (1) Temporal domain: including inspiratory/expiratory phase duration (tPhase), time constants (Tau), and inspiratory-to-expiratory time ratios (TI/TE); (2) airflow domain: comprising mean flow, peak flow, and instantaneous flow at 25%, 50%, and 75% of tidal volume; and (3) spatial domain: quantifying global and regional tidal impedance variation (TIV), global inhomogeneity (GI), and center of ventilation (CoV). Extreme Gradient Boosting (XGBoost) classifiers were trained using 5 distinct data sources (Spirometry, Nasometry, Inspiratory-EIT, Expiratory-EIT, and fused ST-EIT). Model performance was rigorously evaluated via stratified 5-fold cross-validation, and Shapley additive explanations (SHAP) were employed to quantify global and local feature contributions. ResultsThe CP group exhibited a distinct respiratory phenotype compared to controls. In the temporal domain, CP patients showed significantly shorter inspiratory (1.60 s vs.1.85 s, P<0.001) and expiratory phase durations (2.45 s vs. 3.95 s, P<0.001), indicating a rapid, shallow breathing rhythm. In the airflow domain, while inspiratory flows were comparable, the CP group demonstrated significantly elevated mean and peak flows during the expiratory phase (P<0.001), reflecting compensatory respiratory effort. Spatially, CP patients presented significant ventilation redistribution, characterized by higher regional TIV in the right-anterior (ROI1) and left-posterior (ROI4) quadrants, but lower TIV in the left-anterior (ROI2) quadrant. In terms of diagnostic accuracy, the multi-modal ST-EIT model achieved the highest performance (AUC: 0.915±0.012, Accuracy: 0.843±0.019, F1-score: 0.872±0.017), substantially outperforming models based on spirometry (AUC: 0.721) or nasometry (AUC: 0.625) alone. Interpretability analysis revealed that spatial domain features were the most critical, contributing 53.4% to the model’s decision-making, followed by temporal (25.0%) and airflow (21.6%) features. ConclusionST-EIT successfully captures the temporal, airflow, and spatial deviations in CP speech respiration that are undetectable by conventional methods—specifically, rapid phase transitions, hyperdynamic expiratory airflow, and regional ventilation heterogeneity. This study validates ST-EIT as a robust, non-invasive, and radiation-free tool for characterizing speech-respiratory dysfunction, offering high clinical value for bedside screening, rehabilitation planning, and longitudinal monitoring of patients with cleft palate.
3.The Neural Circuit Characteristics of Repetitive Transcranial Magnetic Stimulation Over The Dorsolateral Prefrontal Cortex for The Treatment of Migraine
Chen-Xia JIN ; Bo-Lin TAN ; Yang YE ; Ji-Qing HE ; Ling-Yan WANG ; Zhong-Ming GAO ; Yu-Jun WANG ; Hui-Li LIU ; Yong-Xing YAN ; Xian-Wei CHE
Progress in Biochemistry and Biophysics 2026;53(7):1953-1968
ObjectiveMigraine is a leading neurological disorder and the fourth most common cause of years lived with disability worldwide, affecting nearly 116 million individuals. Although pharmacological treatments are available, their efficacy is often limited by side effects and variable response rates. Repetitive transcranial magnetic stimulation (rTMS) over the dorsolateral prefrontal cortex (DLPFC) offers a safe, non-invasive alternative for migraine management. However, the neurophysiological mechanisms, particularly how rTMS modulates local cortical excitability and distributed pain-related circuits, remain poorly understood. Elucidating these mechanisms is essential for optimizing treatment protocols and improving clinical outcomes. MethodsThis study employed concurrent transcranial magnetic stimulation and electroencephalography (TMS-EEG) to investigate neuroplastic and neurocircuitry mechanisms of DLPFC-rTMS in migraine. Study 1 compared 30 migraineurs and 28 healthy controls to identify abnormalities in TMS-evoked potentials (TEPs) and significant current density (SCD) within sensory-discriminative regions including the primary somatosensory cortex (S1) and posterior insula (pINS), cognitive-affective regions including the anterior insula (aINS) and midcingulate cortex (MCC), and a descending modulatory region, the periaqueductal gray (PAG). Study 2 used a single-blind, crossover, sham-controlled design in 34 healthy participants. Each participant received both active (10 Hz, 80% RMT, 1 500 pulses) and sham DLPFC-rTMS in counterbalanced order. TMS-EEG and cold pain tolerance were assessed before and after each session. ResultsIn Study 1, migraineurs showed a significantly less negative N120 amplitude compared to healthy controls (P=0.027, Cohen’s d=0.60), indicating local intracortical disinhibition. No group differences were observed for N40, P60, or P180 components. At the source level, migraineurs exhibited significantly higher SCD in the S1, pINS, aINS, and MCC (allQ<0.05), but not in the ventroposterior thalamus (vpTHAL), mediodorsal thalamus (mdTHAL), or PAG. In Study 2, active rTMS significantly reduced SCD from pre- to post-stimulation in the S1, aINS, and MCC (all Q<0.05). Sham stimulation also reduced SCD in the S1 (Q<0.05) but not in the aINS or MCC. Although no significant group-level analgesic effect was observed between active and sham conditions (P=0.107), correlation analyses revealed that greater SCD reductions in the S1 and MCC were significantly associated with higher post-rTMS pain tolerance (R=-0.487 and -0.495, both Q<0.01) and larger improvements in pain tolerance(R=-0.487 and -0.451, both Q<0.05). No such correlations were found following sham stimulation, suggesting that the behavioural relevance of neural changes is specific to active rTMS. ConclusionThis study provides novel evidence that migraineurs exhibit both local neuroplastic abnormalities (reduced N120 amplitude) and hyperactivity in key pain-processing regions (S1, pINS, aINS, MCC). A single session of DLPFC-rTMS reduced hyperactivity in the aINS, MCC, and S1. Notably, greater reductions in the S1 and MCC were associated with improved pain tolerance. These findings identify distinct cortical circuitries, particularly within the cognitive-affective pain network, that may serve as potential biomarkers for optimizing rTMS treatment in migraine and other chronic pain conditions. Future studies should validate these results in patient populations experiencing spontaneous migraine attacks and explore multi-session or accelerated rTMS protocols.
4.Construction of risk prediction model for phubbing behavior among college students based on machine learning methods
FU Zheying, LI Yaxin, JIANG Chongming, LI Bo, XU Hui, GE Yang, CHANG Hongjuan
Chinese Journal of School Health 2026;47(7):929-934
Objective:
To develop and compare multiple machine learning models for identifying high-risk college students exhibiting phubbing behavior and to determine key predictive factors, so as to provide evidence for precise screening and early intervention strategies.
Methods:
In December 2025, 1 828 undergraduate students, selected from three universities in Wuhan by using convenience sampling method, were surveyed via online qustionnaire for sociodemographic characteristics, phubbing behavior, family cohesion index, mature happiness, fear of negative evaluation, multidimensional state boredom, and psychological vulnerability. Four machine learning algorithms, including multilayer perceptron (MLP), extreme gradient booting (XGBoost), K-nearest neighbors (KNN), and gradient boosting decision tree (GBDT), were applied. The dataset was randomly split into a training set and a test set at a ratio of 7∶3. Model performance was evaluated using accuracy, recall, F1-score, and area under the receiver operating characteristic curve (AUC). The best performing model was further interpreted using shapley additive explanation (SHAP) analysis to assess feature importance.
Results:
In the training set,955 participants were classified as having low level phubbing behavior, and 325 participants were classified as having high level phubbing behavior. Significant differences were observed in psychological vulnerability, multidimensional state boredom, family cohesion index, and mature happiness between groups ( Z =-10.29, -11.72, -8.17, -7.83, all P <0.05). Significant differences were also found in the detection rate of high level phubbing behavior according to family residence, family status, left behind experience,interpersonal relationships,exercise frequency,physical flexibility,and sleep status ( χ 2=12.22, 38.93, 16.90, 44.64, 58.17, 82.14, 37.89, all P <0.05). Among the four machine learning models (MLP, KNN, GBDT, XGBoost),all showed good discrimination ability, with XGBoost performing best (accuracy 79%, recall 71%, F1-score=0.70, AUC=0.78), while GBDT achieved the highest precision (78%). Feature importance analysis showed that multidimensional state boredom was the most important predictor of phubbing behavior, followed by psychological vulnerability, family cohesion, mature happiness, and fear of negative evaluation. SHAP analysis indicated that multidimensional state boredom, psychological vulnerability, and fear of negative evaluation had positive risk effects, while family cohesion and hedonic well being had protective effects for high risk of phubbing behavior.
Conclusions
Multidimensional state boredom is the core predictor of phubbing behavior among college students. The XGBoost model demonstrates good predictive performance and can provide a reference for identifying high risk individuals and implementing targeted interventions.
5.A qualitative study of self-harm and suicidal behavior in male prisoners
Jiali YANG ; Lan WANG ; Weiyi LIU ; Chao WANG ; Bo YANG ; Hui ZHAO
Chinese Mental Health Journal 2025;39(6):534-540
Objective:To explore the occurrence,co-occurrence,and different subtypes of self-harm and sui-cidal behaviors among male prisoners to inform the development of targeted intervention strategies.Methods:A phenomenology in qualitative research was adopted,and 30 male prisoners with a history of self-harm behaviors,suicidal ideation and suicideattempts,were selected for semi-structured interviews.Data was analyzed by Colaizzi's seven-step method to distill themes.Results:Theme analysis revealed 4 themes and 9 sub-themes(subtypes)of the self-harm and suicidal behaviors of male prisoners,namely the initial occurrence of self-harm behaviors,non-suicid-al self-injury(NSSI)including 6 subtypes(emotional management disorder,self-punishing,antagonistic dissocia-tion,sensation seeking,self-harm without self-awareness,and instrumental self-harm),the co-occurrence of self-harm-suicide behaviors with a subtype of rational suicide,the acute suicide behavior including two subtypes(impul-sive suicide and psychiatric suicide).Conclusion:Negative emotions and emotional dysregulation are important re-lated factorsof self-harm among male prisoners.Suicidal behaviors commonly co-occurred with NSSI,and under-standing the NSSI of prisoners could provide reference for suicide prevention in prisons.There is variability in the motivation of different subtypes of NSSI in maleprisoners,and subtype-based personalized interventions could be a-dopted to reduce the occurrence of self-harm and suicidal behaviors.
6.Resected bone surface anatomy measurement in KBD population for total knee arthroplasty based on three-dimensional computed tomography
Bo YANG ; Ming CHEN ; Shizhang LIU ; Yanhai CHANG ; Hui QIANG ; Ming LING ; Jian TAO ; Pengzhen LEI
Journal of Xi'an Jiaotong University(Medical Sciences) 2025;46(4):650-655
Objective To measure and analyze anatomic parameters of resected distal femurs and proximal tibias in Kaschin-Beck disease(KBD)and osteoarthritic(OA)knees which fit for total knee arthroplasty(TKA),so as to provide basis for designing suitable knee prosthesis for KBD patients.Methods A total of 154 knees(75 KBD and 79 OA)fitting for TKA underwent three-dimensional reconstruction;resection of distal femurs and proximal tibias was simulated during TKA.We measured the distal femoral mediolateral(fML),lateral anteroposterior(fLAP),medial anteroposterior(fMAP),medial condylar width(fMCW),lateral condylar width(fLCW)and fML/fLAP aspect ratio,proximal tibial mediolateral(tML),middle anteroposterior(tAP),medial anteroposterior(tMAP),lateral anteroposterior(tLAP)dimension,and tML/tAP aspect ratio to compare the morphometric differences between KBD and OA knees.Results The measured anatomic parameters of resected distal femurs and proximal tibias in both male and female patients of KBD group were smaller than those of OA knees(P<0.01).We also found that KBD group had larger distal femur fML/fLAP and proximal tibia tML/tAP aspect ratios than those of OA knees(P<0.01).This indicated that under a given AP dimension,KBD knees need an oval-shaped prosthesis with a wider ML length and more spherical-shaped prosthesis with relatively narrow ML dimensions in OA knees.Conclusion The anatomy of distal femurs and proximal tibias shows significant differences in size and shape between KBD and OA knees,which provides theoretical basis for designing proper knee prosthesis for KBD patients.
7.Distalfemur rotational alignment in knee osteoarthritis patients by three-dimensional computed tomography
Bo YANG ; Xianghui HUANG ; Xianghui DONG ; Hui QIANG ; Shizhang LIU ; Pengzhen LEI ; Ming CHEN
Journal of Xi'an Jiaotong University(Medical Sciences) 2025;46(6):991-995
Objective To explore the gender differences and the influence of varus and valgus deformities on distal femur rotational alignment in patients with knee osteoarthritis so as to provide reference for the distal femur rotational osteotomy in total knee arthroplasty(TKA).Methods CT data of 125 knees with osteoarthritis suitable for TKA(including 56 males and 69 females;78 varus knees and 47 valgus knees)were collected and received three-dimensional imaging reconstruction with Mimics software.We measured the posterior condylar angle(PCA),the angle between the anteroposterior axis and the surgical epicondylar axis(ASA),and the angle between the anteroposterior axis and the posterior condylar axis(APA).We also compared gender differences and the influences of varus-valgus deformities on distal femur rotational alignments.Results The PCA and ASA values of distal femur rotational alignments were(3.5±1.1)° vs.(2.8±1.2)°(P=0.001)and(90.6±3.4)° vs.(91.9±3.5)°(P=0.028)for males and females,with statistically differences between the genders.There was no statistically difference in APA between males and females(P=0.482).The PCA values of distal femur rotational alignment in varus and valgus knees were(3.6±1.1)° and(2.9±1.2)°,with a statistically difference between the two groups(P=0.001).There were no statistically differences in ASA or APA between varus and valgus knees(P>0.05).Conclusion Gender and varus-valgus deformities of knee joint affect the distal femur rotational alignment.Distal femur rotational osteotomy angle should be determined according to the patient's CT imaging before TKA to reduce the errors caused by individual differences and improve the accuracy of the distal femur rotational alignment during TKA.
8.Symptoms and quality of life benefits of successful percutaneous coronary intervention in left main disease and/or 3-vessel disease patients with diabetes
Bo-da ZHU ; Tian-tong YU ; Peng HAN ; Bo-hui ZHANG ; Xi ZHANG ; Ping YUAN ; Gang WANG ; Yi YANG ; Hui-li ZHU ; Pan-pan SUN ; Tong-tong LI ; Shuai ZHAO ; Cheng-xiang LI ; Kun LIAN
Chinese Journal of Interventional Cardiology 2025;33(2):93-100
Objective To investigate whether successful percutaneous coronary intervention(PCI)could improve symptoms and quality of life(QOL)in left main disease and/or 3-vessel disease patients with diabetes.Methods Patients with left main disease and/or 3-vessel disease who underwent PCI in the First Affiliated Hospital of Air Force Medical University from April 2018 to May 2021 were consecutively enrolled and subdivided into 2 groups:diabetes and no diabetes.Detailed baseline characteristics,symptoms,including dyspnea and angina,assessed with the Rose dyspnea scale(RDS),Seattle angina questionnaire(SAQ),the European quality of life-5 dimensions(EQ-5D)and 12-item short-form health survey(SF-12)questionnaire respectively,procedural details,and 1 month and 1 year follow-up data were collected.Results Among 440 left main disease and/or 3-vessel disease patients,disease was present in 176(40.00%),who had more hypertension,peripheral artery disease,and LCX lesion(all P<0.05).The incidence of major adverse cardiovascular events(MACE)and all-cause mortality were similar between the two groups(both P>0.05)at 1 month follow-up,while all-cause mortality in diabetes patients was significantly higher than those without diabetes at 1 year follow-up(P=0.013).Low left ventricular ejection fraction was an independent risk factor for MACE and all-cause mortality at 1 month and 1 year follow-up after successful revascularization(all P<0.05).Most importantly,symptoms,including dyspnea and angina,and QOL were markedly improved regardless of diabetes both at 1 month and 1 year follow-up(all P<0.05).Diabetes patients showed improved dyspnea and QOL at similar degree to the non-diabetes patients(all P>0.05)and a more significantly relieved angina(P=0.013).Additionally,the number of chronic total occlusion(CTO)per patient was identified as an independent risk factor of dyspnea(OR 0.723,95%CI 0.525~0.997,P=0.048)and angina relief(OR 0.686,95%CI 0.473~0.995,P=0.047),and the contrast volume(OR 0.995,95%CI 0.992~0.999,P=0.008)as an independent risk factor of QOL improvement in diabetic patients.Conclusions Successful PCI is beneficial for relieving symptoms and improving quality of life in patients with diabetes who have left main disease and/or 3-vessel disease.
9.Association between insulin resistance and idiopathic central precocious puberty in girls and the diagnostic value of insulin resistance
Jin-Bo LI ; Ya XIAO ; Shu-Qin JIANG ; Xiang-Yang LUO ; Hong-Ru ZHANG ; Jun SUN ; Wen-Hui SHI ; Ying YANG ; Wei WANG
Chinese Journal of Contemporary Pediatrics 2025;27(12):1487-1492
Objective To explore the relationship between insulin resistance and idiopathic central precocious puberty(ICPP)in girls and the diagnostic value of insulin resistance.Methods Clinical data of 245 girls aged 4 to 7.5 years with low luteinizing hormone(LH)levels(0.2-0.83 IU/L),normal body weight(body mass index standard deviation score between-2 and+2),and early breast development who visited the Department of Pediatric Endocrinology,Henan Provincial Maternal and Child Health Hospital from January 2022 to March 2025 were retrospectively analyzed.According to the Expert Consensus on the Diagnosis and Treatment of Central Precocious Puberty(2022),patients were assigned to an ICPP group(n=123)or a control group(n=122).Correlations between the homeostasis model assessment of insulin resistance(HOMA-IR)and selected indices were assessed.Multivariable logistic regression was used to evaluate the association between HOMA-IR and ICPP,and the diagnostic performance of various indices for ICPP was evaluated.Results HOMA-IR was higher in the ICPP group than in the control group(P<0.001)and was positively correlated with LH peak(rs=0.467,P<0.05)and the LH peak/FSH peak ratio(rs=0.444,P<0.05).The multivariable logistic regression model including age,BMI,and basal LH showed that HOMA-IR was closely associated with ICPP(OR=2.756,95%CI:1.940-3.913).Receiver operating characteristic curve analysis showed that the areas under the curve for basal LH,HOMA-IR,and their combination in diagnosing ICPP were 0.735,0.735,and 0.805,respectively(P<0.05),and the combined model had a greater area under the curve than either basal LH or HOMA-IR alone(both P<0.05).Conclusions HOMA-IR is closely associated with ICPP in girls with low LH and normal body weight,and combining HOMA-IR with basal LH improves early identification and diagnostic efficiency in this population.
10.Screening and Identification of Nanobodies Against β-Conglycinin
Jia-Shu CHANG ; Hua-Bo SUN ; Yu-Ting WANG ; Xiao-Hui WANG ; Bo YANG ; Hong-Rui LIU ; Yue-Xin LI ; Yuan-Zhao SUN ; Shao-Peng GU ; Jin-Xin HE
Chinese Journal of Biochemistry and Molecular Biology 2025;41(5):764-770
Soy is a vital source of plant carbohydrates.However,it poses significant allergenic risks,particularly to young children and animals.Among the various proteins in soy,β-conglycinin,which con-stitutes approximately 30%of total soy carbohydrates,is a primary allergen.Undigested β-conglycinin can lead to intestinal damage by inhibiting cell growth,disrupting the cytoskeleton,and inducing apopto-sis.It can also enter the lymphatic and circulatory systems,triggering allergic reactions.Conventional ELISA methods for detecting β-conglycinin rely on polyclonal or monoclonal antibodies,which are limited by their large molecular weight,difficulty in accessing the protein core,and sensitivity to acidic and bas-ic conditions.To address these limitations,this study aimed to develop nanobodies(Nbs)against β-con-glycinin.Nbs,derived from the variable regions of heavy-chain antibodies found in camelids,have a mo-lecular weight approximately one-tenth that of conventional antibodies.They offer advantages such as small size,stable structure,high specificity,and strong affinity.A female alpacas was immunized five times using β-conglycinin,which showed a heavy chain antibody potency of 1∶16 000 by ELISA.Pe-ripheral blood lymphocytes were subsequently isolated and total RNA was extracted.The variable region of the heavy-chain antibody was amplified via PCR,and recombinant plasmids were constructed and transformed into the E.coli competency strain ER2738.The resulting library contained about 3.5×108 CFU/mL,which increased to 1.15×1012 PFU/mL after phage rescue,with a 100%Nbs gene insertion rate,indicating high diversity.Its Nbs phage output was significantly enriched by four rounds of solid-phase elution with an enrichment rate of 155.9.Four rounds of solid-phase panning yielded 35 positive clones,all of which shared the same amino acid sequence upon sequencing.The selected Nb was ex-pressed in a prokaryotic system,and its binding ability to β-conglycinin was confirmed using Western blotting and ELISA.The results demonstrated excellent specificity and affinity.This research lays the groundwork for developing a rapid and efficient detection method for β-conglycinin using Nbs,potentially enhancing food safety and allergen management.


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