1.Deep learning model based on fundus images for detection of coronary artery disease with mild cognitive impairment
Yi YE ; Wei FENG ; Yao-dong DING ; Qing CHEN ; Yang ZHANG ; Li LIN ; Tong MA ; Bin WANG ; Xian-gang CHANG ; Zong-yuan GE ; Xiao-yi WANG ; Long-jun CAI ; Yong ZENG
Chinese Journal of Interventional Cardiology 2025;33(6):303-311
Objective To develop a deep learning model based on fundus retinal images to improve the detection rate of mild cognitive impairment(MCI)in patients with coronary heart disease,achieve early intervention and improve prognosis.Methods The study was a single-center cross-sectional study that retrospectively included patients diagnosed with coronary heart disease(CHD)by coronary angiography(≥50% stenosis of at least one coronary vessel)from Beijing Anzhen Hospital between November 2021 and December 2022.The whole data set was randomly divided into the training set and the testing set according to the ratio of 8∶2 for model development.After that,the patient data of the same center from January 2023 to April 2023 were included in the time verification method to verify the model.The diagnostic criteria for MCI were MMSE<27 or MoCA<26.Four kinds of convolutional neural network(CNN)architectures were used to train fundus images,and a comprehensive vision model of MCI detection was established through model integration.The area under the curve(AUC),sensitivity and specificity of the receiver operating curve(ROC)were used to evaluate the performance of the AI model.Results We collected 5 880 eligible fundus images from 3 368 CHD patients.Based on the results of the MMSE scale,the algorithm was labeled,including 2 898 males and 527 MCI patients.The AUC of the deep learning model in the test group is 0.733(95%CI 0.688-0.778),and the sensitivity of the algorithm in the test group is 0.577(95%CI 0.528-0.625)by using the operating point with the maximum sum of sensitivity and specificity.With a specificity of 0.758(95%CI 0.714-0.802),corresponding to a validated AUC of 0.710(95%CI 0.601-0.818).Based on the results of the MoCA scale,the algorithm labels 2 437 males and 1 626 MCI patients.The AUC of the deep learning model in the test group was 0.702(95%CI 0.671-0.733).The operating point with the maximum sum of sensitivity and specificity was selected,and the sensitivity of the algorithm was 0.749(95%CI 0.719-0.778)and the specificity was 0.561(95%CI 0.527-0.595),corresponding to the AUC value of the verification group was 0.674(95%CI 0.622-0.726).Conclusions The deep learning algorithm model based on fundus images has good diagnostic performance,and may be used as a new non-invasive,convenient and rapid screening method for MCI in CHD population.
2.Prokaryotic expression of eIF3i gene encoding protein from lamb testicular cells and the preparation and application of its polyclonal antibody
Qin WU ; Shuaibin WU ; Simei XIAN ; Youci LONG ; Weihao ZHENG ; Mengyi YU ; Jing LI
Chinese Journal of Veterinary Science 2025;45(7):1478-1484
The eIF3i gene was amplified from lamb testicular(LT)cells by PCR and cloned into pCold vector to construct the pCold-eIF3i plasmid.Plasmid PCR,double enzyme digestion and se-quencing were used to verify the results.The recombinant eIF3i protein was induced under the op-timized expression conditions.The expression and reactogenicity of the target protein were detected by SDS-PAGE and Western blot.New Zealand white rabbits were immunized with purified recom-binant eIF3i protein combined with Freund's complete and incomplete adjuvants for three times.Se-rum samples were collected after immunization.Indirect ELISA was used to detect antiserum titer,and Western blot was used to analyze antibody specificity.Indirect immunofluorescence assay(IFA)was used to detect the application effect of antibodies.The results showed that the size of LT eIF3i gene was 978 bp.The optimal expression conditions for the eIF3i recombinant protein were as follows:IPTG concentration of 0.2 mmol/L,temperature of 37 ℃,and induction time of 8 h.The recombinant eIF3i protein was expressed as an inclusion body with a size of about 36 kDa.The titer of polyclonal antibody against eIF3i protein was 1∶51 200.Western blot and IFA showed that the prepared polyclonal antibody against eIF3i protein had good reactivity and specificity.In conclusion,we successfully prepared rabbit anti-eif3i polyclonal antibody and confirmed that it could specifically recognize endogenous eIF3i protein,which laid a foundation for further study on the biological function of eIF3i protein.
3.Establishment and application of RPA-LFD detection method for Pasteurella multocida from duck
Youci LONG ; Qinglin GU ; Simei XIAN ; Weihao ZHENG ; Qin WU ; Mengyi YU ; Jing LI ; Shuaibin WU
Chinese Journal of Veterinary Science 2025;45(3):466-472
This study aims to establish a convenient,new and visual detection method for the field diagnosis of Pasteurella multocida(Pm).With reference to the Pm kmt1 gene conserved sequence published in GenBank,PCR amplification primers were designed,the amplified kmt1 gene was cloned into pMD19-T vector,and the recombinant plasmid standard pMD19-T-kmt 1 was estab-lished and identified by PCR and sequencing.Using pMD1 9-T-kmt 1 plasmid as template and kmt1 gene as target gene,basic primers were designed and synthesized.According to the requirements of LFD,a probe(Pm-P)was designed,and the RPA-LFD method for Pm detection was established by optimizing the reaction conditions.Specificity and sensitivity tests were carried out,and 64 clini-cal samples were tested by the method.The results showed that the established Pm RPA-LFD method could be amplified at 37 ℃ for 15 min.Escherichia coli(E.coli),Salmonella enteriditis(SE),Riemerella anatipestifer(RA),Staphylococcus,goose parvovirus(GPV),duck plague virus(DPV),Muscovy duck parvovirus(MDPV)DNA was extracted as the template,and plasmid standard pMD19-T-kmt 1 was used as the positive control.All the positive controls were negative,indicating that the method had good specificity.The plasmid standard pMD1 9-T-kmt 1 was diluted with a 10-fold ratio,and the plasmid standard with a concentration of 107-100 copies/μL was used as the template.The sensitivity was 1.50×101 copies/μ,,which was 100 times higher than that of PCR.A total of 64 clinical samples with suspected RA were subjected to testing using PCR,RPA and LAMP-LFD,with a 100%compliance rate for all three detection tests.The results show that the established RPA-LFD method has the characteristics of strong specificity,high sensitivity,fast speed and visualization,and can be applied to the field detection of Pm.
4.Deep learning model based on fundus images for detection of coronary artery disease with mild cognitive impairment
Yi YE ; Wei FENG ; Yao-dong DING ; Qing CHEN ; Yang ZHANG ; Li LIN ; Tong MA ; Bin WANG ; Xian-gang CHANG ; Zong-yuan GE ; Xiao-yi WANG ; Long-jun CAI ; Yong ZENG
Chinese Journal of Interventional Cardiology 2025;33(6):303-311
Objective To develop a deep learning model based on fundus retinal images to improve the detection rate of mild cognitive impairment(MCI)in patients with coronary heart disease,achieve early intervention and improve prognosis.Methods The study was a single-center cross-sectional study that retrospectively included patients diagnosed with coronary heart disease(CHD)by coronary angiography(≥50% stenosis of at least one coronary vessel)from Beijing Anzhen Hospital between November 2021 and December 2022.The whole data set was randomly divided into the training set and the testing set according to the ratio of 8∶2 for model development.After that,the patient data of the same center from January 2023 to April 2023 were included in the time verification method to verify the model.The diagnostic criteria for MCI were MMSE<27 or MoCA<26.Four kinds of convolutional neural network(CNN)architectures were used to train fundus images,and a comprehensive vision model of MCI detection was established through model integration.The area under the curve(AUC),sensitivity and specificity of the receiver operating curve(ROC)were used to evaluate the performance of the AI model.Results We collected 5 880 eligible fundus images from 3 368 CHD patients.Based on the results of the MMSE scale,the algorithm was labeled,including 2 898 males and 527 MCI patients.The AUC of the deep learning model in the test group is 0.733(95%CI 0.688-0.778),and the sensitivity of the algorithm in the test group is 0.577(95%CI 0.528-0.625)by using the operating point with the maximum sum of sensitivity and specificity.With a specificity of 0.758(95%CI 0.714-0.802),corresponding to a validated AUC of 0.710(95%CI 0.601-0.818).Based on the results of the MoCA scale,the algorithm labels 2 437 males and 1 626 MCI patients.The AUC of the deep learning model in the test group was 0.702(95%CI 0.671-0.733).The operating point with the maximum sum of sensitivity and specificity was selected,and the sensitivity of the algorithm was 0.749(95%CI 0.719-0.778)and the specificity was 0.561(95%CI 0.527-0.595),corresponding to the AUC value of the verification group was 0.674(95%CI 0.622-0.726).Conclusions The deep learning algorithm model based on fundus images has good diagnostic performance,and may be used as a new non-invasive,convenient and rapid screening method for MCI in CHD population.
5.Comparative study of single-criteria and multi-criteria optimization modes for cervical cancer VMAT radiotherapy plans by Raystation planning system
Gang LI ; Yu-song LONG ; Jun-wen TAN ; Xian-tao HE ; Yong-fu FENG ; Zhan-yu WANG
Chinese Medical Equipment Journal 2025;46(9):39-44
Objective To carry out cervical cancer VMAT radiotherapy planning respectively with the single-criteria optimization(SCO)and multi-criteria optimization(MCO)modes of Raystation 4.7 planning system,and to provide references for selecting optimization mode clinically by comparing and analyzing the dosimetric parameters such as target dose distribution,exposure dose to organ at risk(OAR),monitor unit and beam-on time.Methods Ten cervical cancer patients who attended some hospital from February to December 2022 were retrospectively selected,and some VMAT plans were designed for them with the SCO mode of Raystation 4.7 planning system and then enrolled into a SCO group;other VMAT plans were redesigned with the MCO mode under the premise the setup conditions such as machine model and shot field angle were unchanged,and divided into a MCO group.The two groups were compared in terms of maximum dose(D2%),minimum dose(D98%),homogeneity index(HI)and conformity index(CI)of the planning target volume(PTV),beam-on time,monitor unit and exposure doses to OARs including bladder V50 Gy,V40 Gy and V30 Gy,rectum V50 Gy,V40 Gy and V30 Gy,small intestine V50 Gy,V40 Gy and V30 Gy and left and right femur V50 Gy,V40 Gy and V30 Gy.SPSS 22.0 software was used for statistical analysis.Results Both the two groups met clinical requirements.There were no significant differences between the two groups in D98%,CI,small intestine V50 Gy and left and right femur V40 Gy(P>0.05).The MCO group had the values of HI,D2%,V50 Gy,V40 Gy and V30 Gy,rectum V50 Gy,V40 Gy and V30 Gy,small intestine V40 Gy and V30 Gy and left and right femur V30 Gy lower than those of the SCO group,with the differences being statistically significant(P<0.05).The SCO group had less monitor units and shorter beam-on time when compared with the MCO group,with the differences being statistically siginificant(P<0.05).Conclusion When compared with the SCO mode-based VMAT plans,the MCO mode-based VMAT plans significantly decrease the exposure doses and volume to rectum,bladder and small intestine and raise the PTV homogeneity,while lower the treatment efficiency to some extent by increased monitor units and prolonged beam-on time.[Chinese Medical Equipment Journal,2025,46(9):39-44]
6.Comparative study of single-criteria and multi-criteria optimization modes for cervical cancer VMAT radiotherapy plans by Raystation planning system
Gang LI ; Yu-song LONG ; Jun-wen TAN ; Xian-tao HE ; Yong-fu FENG ; Zhan-yu WANG
Chinese Medical Equipment Journal 2025;46(9):39-44
Objective To carry out cervical cancer VMAT radiotherapy planning respectively with the single-criteria optimization(SCO)and multi-criteria optimization(MCO)modes of Raystation 4.7 planning system,and to provide references for selecting optimization mode clinically by comparing and analyzing the dosimetric parameters such as target dose distribution,exposure dose to organ at risk(OAR),monitor unit and beam-on time.Methods Ten cervical cancer patients who attended some hospital from February to December 2022 were retrospectively selected,and some VMAT plans were designed for them with the SCO mode of Raystation 4.7 planning system and then enrolled into a SCO group;other VMAT plans were redesigned with the MCO mode under the premise the setup conditions such as machine model and shot field angle were unchanged,and divided into a MCO group.The two groups were compared in terms of maximum dose(D2%),minimum dose(D98%),homogeneity index(HI)and conformity index(CI)of the planning target volume(PTV),beam-on time,monitor unit and exposure doses to OARs including bladder V50 Gy,V40 Gy and V30 Gy,rectum V50 Gy,V40 Gy and V30 Gy,small intestine V50 Gy,V40 Gy and V30 Gy and left and right femur V50 Gy,V40 Gy and V30 Gy.SPSS 22.0 software was used for statistical analysis.Results Both the two groups met clinical requirements.There were no significant differences between the two groups in D98%,CI,small intestine V50 Gy and left and right femur V40 Gy(P>0.05).The MCO group had the values of HI,D2%,V50 Gy,V40 Gy and V30 Gy,rectum V50 Gy,V40 Gy and V30 Gy,small intestine V40 Gy and V30 Gy and left and right femur V30 Gy lower than those of the SCO group,with the differences being statistically significant(P<0.05).The SCO group had less monitor units and shorter beam-on time when compared with the MCO group,with the differences being statistically siginificant(P<0.05).Conclusion When compared with the SCO mode-based VMAT plans,the MCO mode-based VMAT plans significantly decrease the exposure doses and volume to rectum,bladder and small intestine and raise the PTV homogeneity,while lower the treatment efficiency to some extent by increased monitor units and prolonged beam-on time.[Chinese Medical Equipment Journal,2025,46(9):39-44]
7.Application of MRI diaphragmatic navigation technology combined with 3D LAVA-FLEX sequence in abdominal enhanced imaging of infants and young children
Di GUO ; Qian-cheng LI ; Cheng-long LI ; Shi-xian LI ; Li-ya LU ; Shu-juan WANG ; Chang-chang LIU ; Xiu-hong DAI
Journal of Regional Anatomy and Operative Surgery 2025;34(10):896-899
Objective To explore the application value of MRI diaphragmatic navigation technology combined with three dimensional liver acquisition with volume acceleration-flexible(3D LAVA-FLEX)sequence in abdominal enhanced imaging of infants and young children.Methods A retrospective analysis was conducted on imaging data of 84 infants and young children who underwent abdominal enhanced MRI examination in our hospital between January 2021 and December 2023.All 84 infants and young children initially underwent conventional dynamic contrast-enhanced 3D LAVA-FLEX sequence scanning;the delayed phase images obtained were included in the dynamic enhancement group.Subsequently,diaphragmatic navigation combined with 3D LAVA-FLEX sequence examination was implemented,and the obtained images were included in the diaphragm navigation group.Subjective scoring was performed for images in both groups,while the signal to noise ratio(SNR),contrast to noise ratio(CNR),and artifact quantification(AQ)were measured and compared between the two groups.Results The respiratory motion artifacts,the clarity of liver parenchyma enhancement,the clarity of liver vascular enhancement,the clarity of spleen parenchyma enhancement and the overall image quality score in the diaphragm navigation group were higher than those in the dynamic enhancement group,and the differences were statistically significant(P<0.05).There were statistically significant differences in SNR and AQ between the two groups of images(P<0.000 1),while there was no statistically significant difference in CNR between the two groups of images(P>0.05).Conclusion Diaphragmatic navigation technology combined with 3D LAVA-FLEX sequence imaging can improve the image quality of abdominal MRI enhanced imaging in infants and young children,and provide a reference for clinical diagnosis and treatment.
8.Prokaryotic expression of eIF3i gene encoding protein from lamb testicular cells and the preparation and application of its polyclonal antibody
Qin WU ; Shuaibin WU ; Simei XIAN ; Youci LONG ; Weihao ZHENG ; Mengyi YU ; Jing LI
Chinese Journal of Veterinary Science 2025;45(7):1478-1484
The eIF3i gene was amplified from lamb testicular(LT)cells by PCR and cloned into pCold vector to construct the pCold-eIF3i plasmid.Plasmid PCR,double enzyme digestion and se-quencing were used to verify the results.The recombinant eIF3i protein was induced under the op-timized expression conditions.The expression and reactogenicity of the target protein were detected by SDS-PAGE and Western blot.New Zealand white rabbits were immunized with purified recom-binant eIF3i protein combined with Freund's complete and incomplete adjuvants for three times.Se-rum samples were collected after immunization.Indirect ELISA was used to detect antiserum titer,and Western blot was used to analyze antibody specificity.Indirect immunofluorescence assay(IFA)was used to detect the application effect of antibodies.The results showed that the size of LT eIF3i gene was 978 bp.The optimal expression conditions for the eIF3i recombinant protein were as follows:IPTG concentration of 0.2 mmol/L,temperature of 37 ℃,and induction time of 8 h.The recombinant eIF3i protein was expressed as an inclusion body with a size of about 36 kDa.The titer of polyclonal antibody against eIF3i protein was 1∶51 200.Western blot and IFA showed that the prepared polyclonal antibody against eIF3i protein had good reactivity and specificity.In conclusion,we successfully prepared rabbit anti-eif3i polyclonal antibody and confirmed that it could specifically recognize endogenous eIF3i protein,which laid a foundation for further study on the biological function of eIF3i protein.
9.Engineering CHO Cell Lines to Stably Express B4GALT1,ST6GAL1,and GnTⅢ with Site-directed Integration
Xian-Hong LI ; Run-Qing JIA ; You-Liang WANG ; Wei-Ling MAN ; Tian-Hao ZHU ; Xin-Long YAN ; Yan-Li LIN
Chinese Journal of Biochemistry and Molecular Biology 2025;41(4):576-585
Glycoengineering was carried out in the mammalian cell line CHO for the production of pro-tein-based drugs.Firstly,the genome sequence of the Rosa26 locus of CHO cells was determined,the gRNA sequences were designed,and the landing pad was integrated into the Rosa26 locus of CHO cells by CRISPR/Cas9 technology.Three targeting vectors co-expressed by glycosyltransferases,which are β-1,4 galactosyltransferase(B4GALT1),α-2,6-sialyltransferase 1(ST6GAL1)and N-acetaminoglycosyl-transferase Ⅲ(GnT Ⅲ),were constructed by overlapping PCR and seamless ligation technology,and the three glycosyltransferase genes were integrated into the CHO Rosa26 locus by Cre enzyme-mediated cassette exchange technology.PCR confirmed that three glycosyltransferases had been successfully site-directed integrated into the Rosa26 site.The mRNA expression levels of the three glycosyltransferases were more than 50 000-fold by qRT-PCR,and the protein expression levels of the three glycosyltrans-ferases were more than 4-fold via western blotting(P<0.001).A CHO-engineered cell line with three glycosyltransferases integrated into Rosa26 site was successfully constructed.
10.Application of MRI diaphragmatic navigation technology combined with 3D LAVA-FLEX sequence in abdominal enhanced imaging of infants and young children
Di GUO ; Qian-cheng LI ; Cheng-long LI ; Shi-xian LI ; Li-ya LU ; Shu-juan WANG ; Chang-chang LIU ; Xiu-hong DAI
Journal of Regional Anatomy and Operative Surgery 2025;34(10):896-899
Objective To explore the application value of MRI diaphragmatic navigation technology combined with three dimensional liver acquisition with volume acceleration-flexible(3D LAVA-FLEX)sequence in abdominal enhanced imaging of infants and young children.Methods A retrospective analysis was conducted on imaging data of 84 infants and young children who underwent abdominal enhanced MRI examination in our hospital between January 2021 and December 2023.All 84 infants and young children initially underwent conventional dynamic contrast-enhanced 3D LAVA-FLEX sequence scanning;the delayed phase images obtained were included in the dynamic enhancement group.Subsequently,diaphragmatic navigation combined with 3D LAVA-FLEX sequence examination was implemented,and the obtained images were included in the diaphragm navigation group.Subjective scoring was performed for images in both groups,while the signal to noise ratio(SNR),contrast to noise ratio(CNR),and artifact quantification(AQ)were measured and compared between the two groups.Results The respiratory motion artifacts,the clarity of liver parenchyma enhancement,the clarity of liver vascular enhancement,the clarity of spleen parenchyma enhancement and the overall image quality score in the diaphragm navigation group were higher than those in the dynamic enhancement group,and the differences were statistically significant(P<0.05).There were statistically significant differences in SNR and AQ between the two groups of images(P<0.000 1),while there was no statistically significant difference in CNR between the two groups of images(P>0.05).Conclusion Diaphragmatic navigation technology combined with 3D LAVA-FLEX sequence imaging can improve the image quality of abdominal MRI enhanced imaging in infants and young children,and provide a reference for clinical diagnosis and treatment.

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