1.Application of immunoprecipitation-mass spectrometry for screening of antibody-recognizing molecules against Echinococcus granulosus
Xiaoxia WU ; Jing DING ; Xuemin JIN ; Zhuangzhi ZHANG ; Lixiao ZHANG ; Mingyuan LIU
Chinese Journal of Veterinary Science 2025;45(3):519-526
We collected the positive serum of Echinococcus granulosus infection in sheep,an inter-mediate host with strong immune response,and used healthy serum as negative control,purified the serum and target protein to capture and enrich the corresponding antigen by immunoprecipita-tion,and obtained target protein-antibody-target protein complex.Mass spectrometry strategies were combined to screen and identify specific antigens associated with Echinococcus granulosus,and the proteins with the highest peptide coverage were analyzed bioinformatically using online prediction software.The results showed that 133 Echinococcus granulosus related proteins were i-dentified by IP-MS.Among them,one protein with peptide coverage≥70%was actin Ⅱ,and three proteins with peptide coverage between 30%to 40%were Ton B box domain containing protein,NADH dehydrogenase(ubiquinone)1 α-subcomplex 2(NADH dehydrogenase[ubiquinone])and lactic dehydrogenase.There were six proteins with 20%to 30%peptide coverage,namely,spli-cing factor 3B subunit 5,tumor protein D52,expressed conserved protein,NADH dehydrogenase(ubiquinone)1 alpha subcomplex 7,inosine-5'-monophosphate dehydrogenase,and aldo keto re-ductase family 1 member B4.Bioinformatics analysis revealed that actin protein has no signal pep-tide,it is probably a non-secretory protein and is subcellularly localized to the cytoskeleton,six op-timal potential antigenic epitopes are present,and the secondary and tertiary structures are consist-ently dominated by α-helices and irregular convolutions.The results indicate that immunoprecipita-tion-mass spectrometry is a high-throughput,simple,rapid and effective method for screening and identifying fine-grained Echinococcus granulosus antigens,which can provide a basis for screening specific molecules for serodiagnostic markers in intermediate host sheep and for the development of novel diagnostic techniques for hydatid diseases.
2.Selection of fecal antigen diagnostic markers for Echinococcus granulosus
Wanli BAN ; Shuai LIU ; Bingjie WANG ; Kamali WULIJIANG ; Xingyu PAN ; Yan WANG ; Talipuhan GU-LIZHATI ; Jing XU ; Teliewuhan MUNILA ; Zhuangzhi ZHANG ; Li ZHAO
Chinese Journal of Veterinary Science 2025;45(4):717-723
This study aims to screen the diagnostic biomarkers for fecal antigen of Echinococcus granulosus(E.granulosus)in dogs with high specificity and sensitivity.The sheep-derived EgPSC artificially infected dogs were collected,and the negative and positive fecal samples of dogs with E.granulosus were prepared by arecoline hydrobromide leakage method.Polyclonal antibody,negative fecal antigen-polyclonal antibody conjugates and positive fecal antigen-polyclonal antibody conju-gates were purified by ammonium sulfate precipitation and affinity chromatography,three groups of samples were detected by ELISA and Western blot,LC-MS/MS and bioinformatics analysis were performed on the three groups of samples.The positive fecal antigen-polyclonal antibody con-jugate was used as the treatment group,the polyclonal antibody and the negative fecal antigen-polyclonal antibody conjugates were used as the control groups to screen the unique peptides of the treatment group.ELISA and Western blot showed that only the positive fecal antigen-polyclonal antibody conjugates were positive.According to LC-MS/MS and bioinformatics analysis,11 unique peptides were screened out only in the treatment group.Among them,3 proteins were related to E.granulosus,namely dysferlin,integrator complex 9 and diagnostic antigen gp50,which were mem-brane-associated proteins,INT complex components and diagnostic antigens.This study has pre-liminarily screened out three candidate canine E.granulosus fecal antigen diagnostic markers,pro-viding a reference for further exploration of diagnostic standards for E.granulosus,screening of echinococcosis target genes,and vaccine development.
3.Application of immunoprecipitation-mass spectrometry for screening of antibody-recognizing molecules against Echinococcus granulosus
Xiaoxia WU ; Jing DING ; Xuemin JIN ; Zhuangzhi ZHANG ; Lixiao ZHANG ; Mingyuan LIU
Chinese Journal of Veterinary Science 2025;45(3):519-526
We collected the positive serum of Echinococcus granulosus infection in sheep,an inter-mediate host with strong immune response,and used healthy serum as negative control,purified the serum and target protein to capture and enrich the corresponding antigen by immunoprecipita-tion,and obtained target protein-antibody-target protein complex.Mass spectrometry strategies were combined to screen and identify specific antigens associated with Echinococcus granulosus,and the proteins with the highest peptide coverage were analyzed bioinformatically using online prediction software.The results showed that 133 Echinococcus granulosus related proteins were i-dentified by IP-MS.Among them,one protein with peptide coverage≥70%was actin Ⅱ,and three proteins with peptide coverage between 30%to 40%were Ton B box domain containing protein,NADH dehydrogenase(ubiquinone)1 α-subcomplex 2(NADH dehydrogenase[ubiquinone])and lactic dehydrogenase.There were six proteins with 20%to 30%peptide coverage,namely,spli-cing factor 3B subunit 5,tumor protein D52,expressed conserved protein,NADH dehydrogenase(ubiquinone)1 alpha subcomplex 7,inosine-5'-monophosphate dehydrogenase,and aldo keto re-ductase family 1 member B4.Bioinformatics analysis revealed that actin protein has no signal pep-tide,it is probably a non-secretory protein and is subcellularly localized to the cytoskeleton,six op-timal potential antigenic epitopes are present,and the secondary and tertiary structures are consist-ently dominated by α-helices and irregular convolutions.The results indicate that immunoprecipita-tion-mass spectrometry is a high-throughput,simple,rapid and effective method for screening and identifying fine-grained Echinococcus granulosus antigens,which can provide a basis for screening specific molecules for serodiagnostic markers in intermediate host sheep and for the development of novel diagnostic techniques for hydatid diseases.
4.Selection of fecal antigen diagnostic markers for Echinococcus granulosus
Wanli BAN ; Shuai LIU ; Bingjie WANG ; Kamali WULIJIANG ; Xingyu PAN ; Yan WANG ; Talipuhan GU-LIZHATI ; Jing XU ; Teliewuhan MUNILA ; Zhuangzhi ZHANG ; Li ZHAO
Chinese Journal of Veterinary Science 2025;45(4):717-723
This study aims to screen the diagnostic biomarkers for fecal antigen of Echinococcus granulosus(E.granulosus)in dogs with high specificity and sensitivity.The sheep-derived EgPSC artificially infected dogs were collected,and the negative and positive fecal samples of dogs with E.granulosus were prepared by arecoline hydrobromide leakage method.Polyclonal antibody,negative fecal antigen-polyclonal antibody conjugates and positive fecal antigen-polyclonal antibody conju-gates were purified by ammonium sulfate precipitation and affinity chromatography,three groups of samples were detected by ELISA and Western blot,LC-MS/MS and bioinformatics analysis were performed on the three groups of samples.The positive fecal antigen-polyclonal antibody con-jugate was used as the treatment group,the polyclonal antibody and the negative fecal antigen-polyclonal antibody conjugates were used as the control groups to screen the unique peptides of the treatment group.ELISA and Western blot showed that only the positive fecal antigen-polyclonal antibody conjugates were positive.According to LC-MS/MS and bioinformatics analysis,11 unique peptides were screened out only in the treatment group.Among them,3 proteins were related to E.granulosus,namely dysferlin,integrator complex 9 and diagnostic antigen gp50,which were mem-brane-associated proteins,INT complex components and diagnostic antigens.This study has pre-liminarily screened out three candidate canine E.granulosus fecal antigen diagnostic markers,pro-viding a reference for further exploration of diagnostic standards for E.granulosus,screening of echinococcosis target genes,and vaccine development.
5.Construction of artificial intelligence neuroimaging education platform for medical imaging students in clinical practice stage and pre-service resident trainees
Ruili LI ; Zhuangzhi SU ; Miao ZHANG ; Dongdong RONG ; Yi SHAN ; Jie LU
Chinese Journal of Medical Education Research 2025;24(2):150-154
Artificial intelligence technology brings new opportunities for the reform and innovation of medical imaging teaching and training models. We took the lead in building an artificial intelligence neuroimaging education platform. The platform included four modules: imaging case library, intelligent interactive learning, self-test and exercise, and online exam. The platform enables a more flexible and convenient education mode, a precise and individualized training method, which can motivate the enthusiasm and initiative of medical imaging students and resident trainees in learning, promote the rapid integration of theoretical knowledge and clinical practice, and improve the efficiency and quality of residency training.
6.Construction of artificial intelligence neuroimaging education platform for medical imaging students in clinical practice stage and pre-service resident trainees
Ruili LI ; Zhuangzhi SU ; Miao ZHANG ; Dongdong RONG ; Yi SHAN ; Jie LU
Chinese Journal of Medical Education Research 2025;24(2):150-154
Artificial intelligence technology brings new opportunities for the reform and innovation of medical imaging teaching and training models. We took the lead in building an artificial intelligence neuroimaging education platform. The platform included four modules: imaging case library, intelligent interactive learning, self-test and exercise, and online exam. The platform enables a more flexible and convenient education mode, a precise and individualized training method, which can motivate the enthusiasm and initiative of medical imaging students and resident trainees in learning, promote the rapid integration of theoretical knowledge and clinical practice, and improve the efficiency and quality of residency training.
7.Effects of canthaxanthin in increasing reproductive hormone secretion through re-ducing ovarian oxidative stress injury on egg production rate of Huaixiang chickens
Zhuo SUN ; Tingting XIE ; Zhuangzhi ZHAO ; Junwen ZHAO ; Anting ZHANG ; Jiang WU ; Mei XIAO ; Lilong AN
Chinese Journal of Veterinary Science 2024;44(10):2266-2276
A total of 360 healthy 26 week old Huaixiang hens were randomly divided into 10 groups,of which,five groups were reared at normal temperature(NC,NT1,NT2,NT3,NT4)and other five groups were reared at high temperature(HC,HT1,HT2,HT3,HT4),all chickens were feed with 0,4,6,8,10 mg/kg canthaxanthin(CX)respectively.The pre-test period was 2 weeks and the main test period was 9 weeks.The results showed that high temperature reduced the egg pro-duction rate(P<0.05),led to damage and hemorrhage in the ovaries,decreased the activity of su-peroxide dismutase(SOD),glutathione peroxidase(GSH-Px)in the serum and the ovaries(P<0.05),decreased the levels of FSH,LH,P4,and E2(P<0.05),decreased the expression of FSHR,LHR,ER,and PR(P<0.05),and increased the levels of malonaldehyde(MDA)(P<0.05)in the ovaries.Addition of CX increased the egg production rate(P<0.05),increased the activity of serum and ovarian SOD and GSH-Px enzymes(P<0.05),decreased the content of serum and ovarian MDA(P<0.05)),increased the ovarian levels of FSH,LH,P4,E2(P<0.05),and increased the o-varian expression of FSHR,LHR,ER,and PR(P<0.05)in Huaixiang chickens at normal and high temperature environmental conditions,respectively.Among them,the highest egg production rate,the strongest antioxidant capacity of serum and ovarian tissues,the best integrity of ovarian tis-sues,ovarian reproductive hormone secretion and expression of reproductive hormone receptors were observed in Huaixiang chickens when 6 mg/kg of CX was added to the diet under normal temperature and when 8 mg/kg CX was added to the diet under high temperature conditions.The results showed that under normal and high temperature conditions,adding CX to the diet signifi-cantly increased the antioxidant capacity of serum and ovaries,alleviated the damage of ovary,pro-moted the secretion of reproductive hormone and the expression of reproductive hormone recep-tors,and increased egg production rate.Dietary supplemented with CX could not restore the dam-age and repair of ovarian tissue,the secretion of reproductive hormone and receptor to normal tem-perature level.The effects of CX on egg production rate,ovarian tissue structure,reproductive hor-mone and receptor expression in chickens were affected by the environmental temperature.When the egg production rate,antioxidant capacity of ovarian tissues,reproductive hormone levels and receptor expression of Huaixiang chickens were optimal,the amount of CX in high temperature en-vironment was more than that of the dose in normal temperature environment.
8.BMP7 expression in mammalian cortical radial glial cells increases the length of the neurogenic period.
Zhenmeiyu LI ; Guoping LIU ; Lin YANG ; Mengge SUN ; Zhuangzhi ZHANG ; Zhejun XU ; Yanjing GAO ; Xin JIANG ; Zihao SU ; Xiaosu LI ; Zhengang YANG
Protein & Cell 2024;15(1):21-35
The seat of human intelligence is the human cerebral cortex, which is responsible for our exceptional cognitive abilities. Identifying principles that lead to the development of the large-sized human cerebral cortex will shed light on what makes the human brain and species so special. The remarkable increase in the number of human cortical pyramidal neurons and the size of the human cerebral cortex is mainly because human cortical radial glial cells, primary neural stem cells in the cortex, generate cortical pyramidal neurons for more than 130 days, whereas the same process takes only about 7 days in mice. The molecular mechanisms underlying this difference are largely unknown. Here, we found that bone morphogenic protein 7 (BMP7) is expressed by increasing the number of cortical radial glial cells during mammalian evolution (mouse, ferret, monkey, and human). BMP7 expression in cortical radial glial cells promotes neurogenesis, inhibits gliogenesis, and thereby increases the length of the neurogenic period, whereas Sonic Hedgehog (SHH) signaling promotes cortical gliogenesis. We demonstrate that BMP7 signaling and SHH signaling mutually inhibit each other through regulation of GLI3 repressor formation. We propose that BMP7 drives the evolutionary expansion of the mammalian cortex by increasing the length of the neurogenic period.
Animals
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Mice
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Humans
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Ependymoglial Cells/metabolism*
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Hedgehog Proteins/metabolism*
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Ferrets/metabolism*
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Cerebral Cortex
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Neurogenesis
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Mammals/metabolism*
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Neuroglia/metabolism*
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Bone Morphogenetic Protein 7/metabolism*
9.Genetically-engineered "all-in-one" vaccine platform for cancer immunotherapy.
Aihua WU ; Yingzhi CHEN ; Hairui WANG ; Ya CHANG ; Meng ZHANG ; Pengfei ZHAO ; Yisi TANG ; Qin XU ; Zhuangzhi ZHU ; Yang CAO ; Yongzhuo HUANG
Acta Pharmaceutica Sinica B 2021;11(11):3622-3635
An essential step for cancer vaccination is to break the immunosuppression and elicit a tumor-specific immunity. A major hurdle against cancer therapeutic vaccination is the insufficient immune stimulation of the cancer vaccines and lack of a safe and efficient adjuvant for human use. We discovered a novel cancer immunostimulant, trichosanthin (TCS), that is a clinically used protein drug in China, and developed a well-adaptable protein-engineering method for making recombinant protein vaccines by fusion of an antigenic peptide, TCS, and a cell-penetrating peptide (CPP), termed an "all-in-one" vaccine, for transcutaneous cancer immunization. The TCS adjuvant effect on antigen presentation was investigated and the antitumor immunity of the vaccines was investigated using the different tumor models. The vaccines were prepared
10. Homeobox Gene Six3 is Required for the Differentiation of D2-Type Medium Spiny Neurons
Xiaolei SONG ; Haotian CHEN ; Zicong SHANG ; Heng DU ; Zhenmeiyu LI ; Yan WEN ; Guoping LIU ; Dashi QI ; Yan YOU ; Zhengang YANG ; Zhuangzhi ZHANG ; Zhejun XU
Neuroscience Bulletin 2021;37(7):985-998
Medium spiny neurons (MSNs) in the striatum, which can be divided into D1 and D2 MSNs, originate from the lateral ganglionic eminence (LGE). Previously, we reported that Six3 is a downstream target of Sp8/Sp9 in the transcriptional regulatory cascade of D2 MSN development and that conditionally knocking out Six3 leads to a severe loss of D2 MSNs. Here, we showed that Six3 mainly functions in D2 MSN precursor cells and gradually loses its function as D2 MSNs mature. Conditional deletion of Six3 had little effect on cell proliferation but blocked the differentiation of D2 MSN precursor cells. In addition, conditional overexpression of Six3 promoted the differentiation of precursor cells in the LGE. We measured an increase of apoptosis in the postnatal striatum of conditional Six3-knockout mice. This suggests that, in the absence of Six3, abnormally differentiated D2 MSNs are eliminated by programmed cell death. These results further identify Six3 as an important regulatory element during D2 MSN differentiation.

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