1.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.
2.Regulation of white adipose tissue in mice by immunization with recombinant Bacillus Calmette-Gue?rin with c-di-AMP adjuvant
Meng-juan DONG ; Yu-xiao CHANG ; Huan-huan NING ; Yan-zhi LU ; Jian KANG ; Ming-ze XU ; Ting DAI ; Jia-ling LI ; Le-ran HAO ; Lin-na ZHANG ; Yin-lan BAI
Chinese Journal of Zoonoses 2025;41(4):370-375
This study assessed the role and mechanism of the recombinant Bacillus Calmette-Gue?rin vaccine(rBCG)with c-di-AMP adjuvant in regulating metabolism and immunity in epididymal white adipose(eWAT)in mice.Male C57BL/6 mice were intravenously immunized with BCG and rBCG,and their body weights were monitored.eWAT was isolated from the mice,and the stromal vascular fractions(SVFs)cell number was counted with a hemocytometer.Sections of mouse adipose tissue were prepared,and the size,number,and morphology of eWAT adipocytes and crown-like structure(CLS)formation were compared under a microscope after HE staining.The transcription levels of lipid metabolism-associated factors,cytokines and aging-associated genes in each group were determined with qRT-PCR.The body weights of mice gradually increased after immunization with BCG and rBCG.The proportions of eWAT increased,and the SVFs cell number decreased,in rBCG immunized mice.HE staining indicated that BCG immunization promoted hyperplasia,whereas rBCG immunization promoted hypertrophy of eWAT adipocytes;moreover,both BCG and rBCG immunization induced CLS formation in eWAT.The qRT-PCR results indicated that rBCG immunization inhibited the expression of genes associated with lipolysis and energy expenditure in eWAT.BCG immunization had little effect on cytokine transcription,whereas rBCG significantly induced the transcription of IFN-γ and IL-1Ra,and inhibited that of IL-15 and IL-2,but did not induce the expression of aging-associated genes.Thus,rBCG immunization induced eWAT adipocyte hypertrophy,which was associated with the inhibition of eWAT lipolysis and the regulation of cytokine expression.
3.Astragaloside Ⅳ inhibits LPS-induced RAW 264.7 macrophage polarization and regulates their migration via cGAS/STING/NF-κB pathway
Chang-chao YANG ; Guo-ting LI ; Lin LIU ; Zi-xian ZHAO ; Wei-kang LI ; Qing-xin SUN ; Yu-ying ZHAO ; Jing-shan ZHAO
Chinese Pharmacological Bulletin 2025;41(7):1290-1297
Aim To explore the effect of astragalosideⅣ(AS-Ⅳ)on lipopolysaccharide(LPS)-induced po-larization and migration of RAW 264.7 macrophages and the underlying mechanism.Methods 1 mg·L-1 LPS was used to construct cell migration model.Scratch assay was utilized to determine cell migration rate.Immunofluorescence staining was utilized to de-tect the expression and location of F4/80,iNOS and Arg-1.CCK-8 assay was used to determine the viabili-ty of RAW 264.7 cells.Griess assay was used to measure NO content.Molecular docking was used to analyze the interaction between AS-Ⅳ and the core tar-gets such as cGAS and STING protein.Western blot was employed to detect the expression of iNOS,Arg-1,cGAS,STING,NF-κB p65 and p-NF-κB p65 protein.Results AS-Ⅳ significantly inhibited the migration and M1 polarization of RAW 264.7 cells induced by LPS.Moreover,AS-Ⅳ could interact with cGAS and STING protein,especially cGAS.Further Western blot assay showed that AS-Ⅳ significantly downregulated the expression of iNOS,cGAS,STING and p-NF-κB p65 protein.Conclusions AS-Ⅳ could promote mac-rophage M1 to M2 polarization,thereby inhibited mac-rophage migration through restraining the cGAS/STING/NF-κB signaling pathway,which provides a new therapeutic target for AS-Ⅳ to improve the early inflammatory response of AS.
4.Ultrasonic Extraction-Single-Particle Inductively Coupled Plasma Mass Spectrometry for Analysis of Metal Nanoparticles in Seawater Sediments
Jie CHUN ; Yu-Mei SONG ; Chang WANG ; Rui-Ling ZHANG ; Peng-Ran GUO ; Wei-Xin LIANG ; Ting ZHANG
Chinese Journal of Analytical Chemistry 2025;53(6):987-997
Accurate analysis of metal nanoparticles(MNPs)in sediments is a prerequisite for assessing the ecological risks of MNPs in aquatic environmental sediments.In this study,an analytical method for quantitative detection of concentration and particle size distribution of silver-containing nanoparticles(Ag-NPs),zinc-containing nanoparticles(Zn-NPs),cerium-containing nanoparticles(Ce-NPs),and titanium-containing nanoparticles(Ti-NPs)in sediments was established based on ultrasonic extraction-single particle inductively coupled plasma mass spectrometry(SP-ICP-MS).The effects of sample preparation conditions such as extraction solvent type,solid-liquid ratio,ultrasonic time,and settling time on the recovery of MNPs were investigated.The results showed that the extraction of MNPs from sediment by distilled water could effectively eliminate the high background signal interference introduced by the extractant under the conditions of solid-liquid ratio of 1∶400(g∶mL),ultrasonic extraction time of 1 h and settling time of 3 h.The detection limits for particle size of Ag-NPs,Zn-NPs,Ce-NPs and Ti-NPs in sediments were 31,35,26 and 85 nm,respectively,while the detection limits of particle concentrations were 1.21×104,1.90×104,5.26×107 and 1.48×107 particles/g,respectively.The spiking recoveries of Ag-NPs,Zn-NPs,Ce-NPs and Ti-NPs in sediments were 62.1%-108.7%,with relative standard deviations below 10%.This method could rapidly,accurately and simultaneously determine the concentration and particle size distribution of various MNPs in sediments,and was successfully applied to analysis of Ag-NPs,Zn-NPs,Ce-NPs,and Ti-NPs in authentic marine sediments.
5.Frequent association of malignant effusions in plasmablastic lymphoma:a single‑institutional experience of nine cases in Taiwan
Bo‑Jung CHEN ; Yu‑Ting KUO ; Sheng‑Tsung CHANG ; Khin‑Than WIN ; Shang‑Wen CHEN ; Sheng‑Yen HSIAO ; Yin‑Hsun FENG ; Yen‑Chuan HSIEH ; Shih‑Sung CHUANG
Blood Research 2025;60():22-
Purpose:
Plasmablastic lymphoma (PBL) is a rare, aggressive lymphoma that is characterized by terminal B-cell differ‑ entiation. In the West, PBL usually occurs in patients with immunodeficiencies, particularly those induced by human immunodeficiency virus (HIV) infection. We investigated the clinicopathological features of PBL at a single institute in Taiwan, where HIV infection is rare.
Methods:
This retrospective chart review identified PBL cases that were treated at a single institute in southern Tai‑ wan between 2008 and 2024.
Results:
We identified nine patients (four males and five females; median age 71 years). Of the eight patients tested for HIV, only one tested positive. Pathologically, the tumors showed plasmablastic morphology and immunopheno‑ type, and three (33%) cases tested positive for Epstein–Barr virus. Six (67%) patients presented with Stage IV disease, including five (56%) with malignant effusion. Six patients were treated with chemotherapy and the remaining three received only supportive care. During a median follow-up of 10 months, five patients died of progressive disease, two died of unrelated diseases, and two were alive with PBL relapse.
Conclusion
In Taiwan, PBL constitutes a rare and aggressive clinical condition and is frequently associated with malignant effusion. In contrast to Western patients, the PBL in most patients from Taiwan was unrelated to HIV infection.
6.In-room cytologic evaluation by trained endosonographer for determination of procedure end in endoscopic ultrasound-guided fine needle biopsy of solid pancreatic lesions: a prospective study in Taiwan
Weng-Fai WONG ; Yu-Ting KUO ; Wern-Cherng CHENG ; Chia-Tung SHUN ; Ming-Lun HAN ; Chieh-Chang CHEN ; Hsiu-Po WANG
Clinical Endoscopy 2025;58(3):465-473
Background/Aims:
Endoscopic ultrasound-guided fine needle biopsy (EUS-FNB) is an essential tool for tissue acquisition in solid pancreatic tumors. Rapid on-site evaluation (ROSE) by cytologists ensures diagnostic accuracy. However, the universal application of the ROSE is limited by its availability. Therefore, we aimed to investigate the feasibility of determining the end of the procedure based on the results of in-room cytological evaluation by trained endosonographers (IRCETE).
Methods:
A training course focusing on the cytological interpretation of common pancreatic tumors was provided to the three endosonographers. After training, the decision to terminate EUS-FNB was made based on IRCETE results. The diagnostic accuracy, concordance rate of diagnostic categories, and sample adequacy were compared with those determined by board-certified cytologists and macroscopic on-site evaluation (MOSE).
Results:
We enrolled 65 patients with solid pancreatic tumors, most of whom were malignant (86.2%). The diagnostic accuracy was 90.8% when the end of the procedure was determined based on IRCETE, compared to 87.7% and 98.5% when determined by MOSE and cytologists, respectively (p=0.060). Based on the cytologists’ results, the accuracy of IRCETE in diagnostic category interpretation was 97.3%.
Conclusions
In the absence of ROSE, IRCETE can serve as a supplementary alternative to MOSE in determining the end of tissue sampling with a high accuracy rate.
7.The relationship between family function and anxiety/depression among medical college students:the mediating role of loneliness
Ying FENG ; Yujing TAO ; Haoqi LI ; Ting YU ; Weiwei CHANG ; Liying WEN
Journal of Shenyang Medical College 2025;27(4):359-363
Objective:To explore the relationship between medical students'family function and anxiety/depression,and the mediating role of loneliness in this relationship.Methods:A total of 577 medical students were surveyed using stratified cluster sampling.Questionnaires assessed family function,loneliness,anxiety,and depression.Spearman correlation analysis was used to examine the relationships among family function,loneliness,anxiety,and depression.The mediating effects were tested using SPSS 26.0 PROCESS program model 4.Results:Medical students'family function,loneliness,anxiety,and depression scores were 8.00(5.00,10.00),16.00(12.00,19.00),3.00(2.00,4.00),and 3.00(2.00,4.00),respectively.Spearman correlation analysis showed that family function was negatively correlated with loneliness(r=-0.337),anxiety(r=-0.237),and depression(r=-0.257)(all P<0.01).Loneliness was positively correlated with anxiety(r=0.394)and depression(r=0.392)(both P<0.01).Anxiety was positively correlated with depression(r=0.746,P<0.01).Family function negatively predicted anxiety(β=-0.118,P<0.01),depression(β=-0.105,P<0.01),and loneliness(β=-0.322,P<0.01).Loneliness positively predicted anxiety(β=0.348,P<0.01)and depression(β=0.346,P<0.01).Loneliness played a partial mediating role in the relationship of family function with anxiety and depression among medical students,with mediating effects of 49.0%and 43.2%,respectively.Conclusions:Family function not only directly affects medical students'anxiety and depression but also indirectly affects them by influencing loneliness.Therefore,improving medical students'family function can effectively reduce their loneliness,thereby helping to alleviate anxiety and depression.
8.Frequent association of malignant effusions in plasmablastic lymphoma:a single‑institutional experience of nine cases in Taiwan
Bo‑Jung CHEN ; Yu‑Ting KUO ; Sheng‑Tsung CHANG ; Khin‑Than WIN ; Shang‑Wen CHEN ; Sheng‑Yen HSIAO ; Yin‑Hsun FENG ; Yen‑Chuan HSIEH ; Shih‑Sung CHUANG
Blood Research 2025;60():22-
Purpose:
Plasmablastic lymphoma (PBL) is a rare, aggressive lymphoma that is characterized by terminal B-cell differ‑ entiation. In the West, PBL usually occurs in patients with immunodeficiencies, particularly those induced by human immunodeficiency virus (HIV) infection. We investigated the clinicopathological features of PBL at a single institute in Taiwan, where HIV infection is rare.
Methods:
This retrospective chart review identified PBL cases that were treated at a single institute in southern Tai‑ wan between 2008 and 2024.
Results:
We identified nine patients (four males and five females; median age 71 years). Of the eight patients tested for HIV, only one tested positive. Pathologically, the tumors showed plasmablastic morphology and immunopheno‑ type, and three (33%) cases tested positive for Epstein–Barr virus. Six (67%) patients presented with Stage IV disease, including five (56%) with malignant effusion. Six patients were treated with chemotherapy and the remaining three received only supportive care. During a median follow-up of 10 months, five patients died of progressive disease, two died of unrelated diseases, and two were alive with PBL relapse.
Conclusion
In Taiwan, PBL constitutes a rare and aggressive clinical condition and is frequently associated with malignant effusion. In contrast to Western patients, the PBL in most patients from Taiwan was unrelated to HIV infection.
9.In-room cytologic evaluation by trained endosonographer for determination of procedure end in endoscopic ultrasound-guided fine needle biopsy of solid pancreatic lesions: a prospective study in Taiwan
Weng-Fai WONG ; Yu-Ting KUO ; Wern-Cherng CHENG ; Chia-Tung SHUN ; Ming-Lun HAN ; Chieh-Chang CHEN ; Hsiu-Po WANG
Clinical Endoscopy 2025;58(3):465-473
Background/Aims:
Endoscopic ultrasound-guided fine needle biopsy (EUS-FNB) is an essential tool for tissue acquisition in solid pancreatic tumors. Rapid on-site evaluation (ROSE) by cytologists ensures diagnostic accuracy. However, the universal application of the ROSE is limited by its availability. Therefore, we aimed to investigate the feasibility of determining the end of the procedure based on the results of in-room cytological evaluation by trained endosonographers (IRCETE).
Methods:
A training course focusing on the cytological interpretation of common pancreatic tumors was provided to the three endosonographers. After training, the decision to terminate EUS-FNB was made based on IRCETE results. The diagnostic accuracy, concordance rate of diagnostic categories, and sample adequacy were compared with those determined by board-certified cytologists and macroscopic on-site evaluation (MOSE).
Results:
We enrolled 65 patients with solid pancreatic tumors, most of whom were malignant (86.2%). The diagnostic accuracy was 90.8% when the end of the procedure was determined based on IRCETE, compared to 87.7% and 98.5% when determined by MOSE and cytologists, respectively (p=0.060). Based on the cytologists’ results, the accuracy of IRCETE in diagnostic category interpretation was 97.3%.
Conclusions
In the absence of ROSE, IRCETE can serve as a supplementary alternative to MOSE in determining the end of tissue sampling with a high accuracy rate.
10.Cognitive function disparities among atrial fibrillation patients with varying comorbidities.
Mei-Qi ZHAO ; Ting SHEN ; Man-Lin ZHAO ; Jia-Xin LIU ; Mei-Lin XU ; Xin LI ; Liu HE ; Yu KONG ; Chang-Sheng MA
Journal of Geriatric Cardiology 2025;22(10):859-870
BACKGROUND:
Mild cognitive impairment (MCI) is common in atrial fibrillation (AF) patients and may develop earlier in those with multiple cardiovascular comorbidities, potentially impairing self-management and treatment adherence. This study aimed to characterize the prevalence and profile of MCI in AF patients, examine its associations with cardiovascular comorbidities, and assess how these comorbidities influence specific cognitive domains.
METHODS:
This cross-sectional study analyzed data from AF patients who underwent cognitive assessment between 2017 and 2021. Cognitive status was categorized as MCI or non-MCI based on the Montreal Cognitive Assessment. Associations between comorbidities and MCI were assessed by logistic regression, and cognitive domains were compared using the Mann-Whitney U test.
RESULTS:
Of 4136 AF patients (mean age: 64.7 ± 9.4 years, 64.7% male), 33.5% of patients had MCI. Among the AF patients, 31.2% of patients had coronary artery disease, 20.1% of patients had heart failure, and 18.1% of patients had hypertension. 88.7% of patients had left atrial enlargement, and 11.0% of patients had reduced left ventricular ejection fraction. Independent factors associated with higher MCI prevalence included older age (OR = 1.04, 95% CI: 1.03-1.05, P < 0.001), lower education level (OR = 1.51, 95% CI: 1.31-1.73, P < 0.001), hypertension (OR = 1.28, 95% CI: 1.07-1.52, P = 0.001), heart failure (OR = 1.24, 95% CI: 1.04-1.48, P = 0.020), and lower left ventricular ejection fraction (OR = 1.43, 95% CI: 1.04-1.98, P = 0.028). A higher CHA2DS2-VASc score (OR = 1.27, 95% CI: 1.22-1.33, P < 0.001; ≥ 2 points vs. < 2 points), and greater atherosclerotic cardiovascular disease burden (OR = 1.45, 95% CI: 1.02-2.08, P = 0.040; 2 types vs. 0 type) were linked to increased MCI risk. These above factors influenced various cognitive domains.
CONCLUSIONS
MCI is common in AF and closely associated with cardiovascular multimorbidity. Patients with multiple comorbidities are at higher risk, highlighting the importance of routine cognitive assessment to support self-management and integrated care.

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