2.Production and identification of PEDV RBD protein peptide antibody
Jiayu YU ; Yuhang JIANG ; Guoqing ZHANG ; Lichao YI ; Shuang ZHANG ; Letian LI ; Aijian QIN ; Chang LI
Chinese Journal of Veterinary Science 2025;45(7):1357-1365
This study aims to screen epitope antigens targeting the receptor binding domain(RBD)of porcine epidemic diarrhea virus(PEDV)based on its amino acid sequence(GenBank accession number:AKN45969.1),prepare PEDV RBD polyclonal antibody,and perform their identification.Bioinformatics analysis software was used to predict the potential antigenic epitopes of PEDV RBD and sequence comparison with porcine coronavirus strains was performed,the selected dominant antigen epitopes were then conjugated with keyhole limpet hemocyanin(KLH),to synthesize pep-tides directly and immunize mice to generate specific antibody,Western blot technique and indirect immunofluorescence assay were utilized to identify the specificity of the antibodies,and indirect ELISA method was further applied to determine the antibody potency.Results showed the selected PEDV RBD dominant epitope sequence shared 100%similarity with 18 other PEDV strains,while exhibiting low sequence similarity with 11 TGEV strains(27.8%—29.3%)and 16 PDCoV strains(10.5%—13.4%),indicating good epitope conservation.Western blot showed that the specificity of the prepared peptide antibody specifically recognized the PEDV RED protein overexpressed in Ex-pi293F cells and overexpressed in baculovirus system,and at the same time,the antibody was still able to detect the PEDV S protein expressed in PEDV-infected Vero cells at a 1∶2 000 dilution,while it did not react with TGEV-and PDCoV-infected ST cells,indicating that the good specificity of the peptide antibody.ELISA revealed that the potency of specific antibodies in mouse serum could reach up to 1∶25 600.The above results indicate that bioinformatics techniques were suc-cessfully utilized to predict antigenic epitopes of PEDV RBD protein,and specific PEDV RBD pep-tide antibodies were prepared.
3.Production and identification of PEDV RBD protein peptide antibody
Jiayu YU ; Yuhang JIANG ; Guoqing ZHANG ; Lichao YI ; Shuang ZHANG ; Letian LI ; Aijian QIN ; Chang LI
Chinese Journal of Veterinary Science 2025;45(7):1357-1365
This study aims to screen epitope antigens targeting the receptor binding domain(RBD)of porcine epidemic diarrhea virus(PEDV)based on its amino acid sequence(GenBank accession number:AKN45969.1),prepare PEDV RBD polyclonal antibody,and perform their identification.Bioinformatics analysis software was used to predict the potential antigenic epitopes of PEDV RBD and sequence comparison with porcine coronavirus strains was performed,the selected dominant antigen epitopes were then conjugated with keyhole limpet hemocyanin(KLH),to synthesize pep-tides directly and immunize mice to generate specific antibody,Western blot technique and indirect immunofluorescence assay were utilized to identify the specificity of the antibodies,and indirect ELISA method was further applied to determine the antibody potency.Results showed the selected PEDV RBD dominant epitope sequence shared 100%similarity with 18 other PEDV strains,while exhibiting low sequence similarity with 11 TGEV strains(27.8%—29.3%)and 16 PDCoV strains(10.5%—13.4%),indicating good epitope conservation.Western blot showed that the specificity of the prepared peptide antibody specifically recognized the PEDV RED protein overexpressed in Ex-pi293F cells and overexpressed in baculovirus system,and at the same time,the antibody was still able to detect the PEDV S protein expressed in PEDV-infected Vero cells at a 1∶2 000 dilution,while it did not react with TGEV-and PDCoV-infected ST cells,indicating that the good specificity of the peptide antibody.ELISA revealed that the potency of specific antibodies in mouse serum could reach up to 1∶25 600.The above results indicate that bioinformatics techniques were suc-cessfully utilized to predict antigenic epitopes of PEDV RBD protein,and specific PEDV RBD pep-tide antibodies were prepared.
4.Quantitative Analysis of Multi-components in Gnaphalium affine Based on UPLC-Q-Exactive Orbitrap MS
Chunyan CAO ; Jiaqi ZHANG ; Zhenduo ZHAO ; Qin SHEN ; Shenglan QI ; Wei LIU ; Lichao ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):148-155
ObjectiveTo establish a rapid and stable liquid chromatography-mass spectrometry(LC-MS) for simultaneous analysis of 17 chemical components in Gnaphalium affine aboveground parts with flowers, so as to provide experimental basis for improving the quality standard of this herb. MethodUltra performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used for the quantitative analysis of 17 constituents in 15 batches of G. affine from different origins, the separation was performed on an ACQUITY UPLC® BEH C18 column(2.1 mm×100 mm, 1.7 μm) with the mobile phase of methanol(A)-0.1% formic acid aqueous solution(B) for gradient elution(0-1.0 min, 8%A; 1.0-4.0 min, 8%-26%A; 4.0-9.0 min, 26%A; 9.0-14.0 min, 26%-34%A; 14.0-14.5 min, 34%-45%A; 14.5-15.0 min, 45%-60%A; 15.0-18.0 min, 60%-90%A; 18.0-19.0 min, 90%A; 19.0-19.01 min, 90%-8%A; 19.01-20.0 min, 8%A), the flow rate was 0.3 mL·min-1, the column temperature was 40 ℃ and the injection volume was 2 μL. And the electrospray ionization was used with full scanning in both positive and negative ion modes, and the scanning range was m/z 100-1 000. ResultThe established method has been verified by the methodology and could be used for the simultaneous quantification of 17 components in G. affine. The content ranges of the 17 components(quinic acid, gallic acid, protocatechuic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 1,3-O-dicaffeoylquinic acid, isochlorogenic acid A, isoquercitrin, 1,5-O-dicaffeoylquinic acid, apigenin-7-O-glucoside, astragalin, isochlorogenic acid C, luteolin, apigenin and hispidulin) in 15 batches of G. affine samples was 39.60-179.12, 0.17-0.84, 2.41-8.38, 4.33-31.50, 13.63-180.38, 2.43-14.75, 1.16-19.68, 0.49-5.63, 55.77-445.16, 0.23-10.26, 62.04-530.10, 1.11-18.01, 11.36-90.61, 12.22-65.98, 7.22-69.84, 3.37-45.65, 0.30-2.59 μg·g-1, respectively. The content of organic acids was higher than that of flavonoids in G. affine, and the contents of 1,5-O-dicaffeoylquinic acid, isochlorogenic acid A, quinic acid and chlorogenic acid were higher. Meanwhile, the content of flavonoids in the samples from Guizhou was higher than that from Jiangsu, while the content of organic acids in the samples from Jiangsu was higher than that from Guizhou. ConclusionThe established method can be used for the rapid and accurate determination of 17 components in G. affine, which clarifies the content range of the main components in this herb, and can provide a reference for the selection of quality control markers of G. affine.
5.Regulation of imagery training on stress response of flying cadets on their first solo flight
Qin ZHAI ; Lichao WANG ; Shuo FAN ; Taiyu LIU ; Wenwen ZHENG ; Yan LI
Journal of Navy Medicine 2024;45(11):1137-1142
Objective To explore the regulatory effect of imagery training on the stress response of flying cadets on their first solo flight.Methods A total of 84 fourth grade flying cadets from an aviation university were randomly assigned to three groups(n=28).The imagery training group used imagery training for stress intervention,the comfort training group received comfort training,and the blank control group did not do any additional training.The heart rate,salivary amylase concentration and stress score before and after the first solo flight and flight performance were recorded.Results During the first solo flight,the average heart rate(P<0.001),stress score(P<0.001)and flight performance(P<0.001)increased in all the cadets.Imagery training could significantly reduce the average heart rate(P<0.01),stress score(P<0.001)on the first solo flight and the salivary amylase concentration after the first solo flight(P<0.01).Conclusion Imagery training can effectively reduce the stress response of flying cadets on their first solo flight.
6.Interdisciplinary teaching-assisted education reform in "Principal Biology".
Lichao SUN ; Xiaoyan MA ; Zhenya CHEN ; Qin ZOU ; Yixin HUO
Chinese Journal of Biotechnology 2023;39(11):4718-4729
General education in biological courses such as "Principal Biology" is an essential avenue for gaining an understanding of life science and developing an interest in the field. The reform of biological education teaching mode based on interdisciplinary approaches aims to foster cross-disciplinary talents, which is crucial for the rapid development of China's bioeconomy. Teaching method that simply superimposes different subjects is difficult to discover the value of interdisciplinary education. To address this, a novel teaching system and an innovative teaching mode were proposed for "Principal Biology" course by integrating science and engineering subjects, based on the cross-disciplinary feature in Beijing Institute of Technology. The system involves the design of cross-disciplinary course content and the integration of multiple disciplines and knowledge points based on students' majors, taking into account the characteristics of students' physical and mental development. To improve students' scientific literacy and interdisciplinary thinking ability, differentiated and major-driven teaching modes were applied by incorporating the "1+N" mixed and immersive cross-thinking training. The effectiveness of tailored cross-disciplinary teaching was evaluated using "in-teaching" and "post-teaching" data feedback models, which promote the optimization of teaching process and enhance the quality of education in cross-disciplinary biological science.
Humans
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Students
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Curriculum
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Biological Science Disciplines/education*
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Universities
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Biology/education*
7.Clinical value of improved sequential organ failure assessment in predicting the prognosis of Klebsiella pneumoniae sepsis
Lei GAO ; Haiying CHEN ; Chong QI ; Lichao QIN ; Yuxiang LI
Chinese Journal of Infectious Diseases 2021;39(1):40-45
Objective:To explore the value of improved sequential organ failure assessment (ISOFA) in predicting the prognosis of Klebsiella pneumoniae sepsis. Methods:The clinical data of 379 patients with Klebsiella pneumoniae sepsis admitted to the First Hospital of Jilin University from January 1, 2018 to June 30, 2019 were retrospectively analyzed.They were divided into survival group and death group according to the 28-day prognosis, and the age, gender, C-reactive protein, procalcitonin, grade of bacterial resistance, and the occurrence of septic shock of the two groups were compared.Statistical analysis was conducted by using independent sample t test, Mann-Whitney U test and chi-square test. Risk factors for the prognosis of the disease was analyzed by logistic regression analysis.The correlation between ISOFA and other scoring system including Charlson′s weighted index of comorbidities (WIC), national early warning system (NEWS), acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), simplified acute physiology scoreⅡ (SAPSⅡ), mortality in emergency department sepsis score (MEDS), sequential organ failure assessment (SOFA), acute gastrointestinal injury (AGI) was determined by Spearman correlation coefficient.The area under the receiver operating characteristic curve (AUROC) was used to compare the evaluation value of each scoring system for the prognosis of Klebsiella pneumoniae sepsis. The clinical significance of ISOFA risk stratification was evaluated by Kaplan-Meier survival curve. Results:Among the 379 patients with Klebsiella pneumoniae sepsis, 278 were in the survival group and 101 were in the death group.The differences of age, gender, the occurrence of septic shock, grade of bacterial resistance, C-reactive protein, procalcitonin, WIC, NEWS, APACHEⅡ, SAPSⅡ, MEDS, SOFA, AGI and ISOFA score between the two groups were all statistically significant ( t=-3.218; χ2=6.781, 24.374 and 27.208, respectively; Z=-3.689, -5.022, -4.396, -4.697, -7.438, -6.348, -6.358, -8.676, -6.680 and -11.658, respectively; all P<0.01). The calculation method of ISOFA was obtained by single factor analysis: ISOFA=SOFA+ 1.5×AGI.Multivariate logistic regression suggested that ISOFA, gender, SOFA, procalcitonin, C-reactive protein, and bacterial resistance were independent risk factors for the prognosis of the disease. The Spearman correlation coefficients of ISOFA and WIC, NEWS, APACHEⅡ, SAPSⅡ, MEDS and SOFA were 0.327, 0.371, 0.614, 0.564, 0.578 and 0.847, respectively. The AUROC and its 95% confidence interval for WIC, NEWS, MEDS, SAPSⅡ, APACHEⅡ, SOFA alone, SOFA plus AGI, and ISOFA to predict the prognosis of Klebsiella pneumoniae sepsis were 0.646 (0.584-0.708), 0.657 (0.597-0.716), 0.712 (0.654-0.771), 0.713 (0.653-0.773), 0.749 (0.693-0.806), 0.788 (0.737-0.838), 0.872 (0.826-0.917) and 0.891 (0.845-0.937), respectively.The results showed that ISOFA had the best predictive effect.The Kaplan-Meier survival curve suggested that there were statistical differences in survival rate among ISOFA low-risk level, medium-risk level, and high-risk level (all P<0.01). Conclusion:ISOFA has important clinical significance in predicting the prognosis of Klebsiella pneumoniae sepsis.
8.Feasibility of nalbuphine for patient-controlled intravenous analgesia after cesarean section
Zairong TANG ; Ze QIN ; Guang LI ; Sirui WU ; Lichao DI
Chinese Journal of Anesthesiology 2021;41(8):975-977
Objective:To preliminarily evaluate the effectiveness and safety of nalbuphine for patient-controlled intravenous analgesia (PCIA) after cesarean section.Methods:This study was a single arm clinical trial.Sixty parturients, aged 20-44 yr, weighing 50-80 kg, of American Society of Anesthesiologists physical status ⅠorⅡ, scheduled for cesarean section with epidural anesthesia, were enrolled in this study.PCIA was performed when visual analogue scale (VAS) score for postoperative pain ≥3 points following cesarean section.A bolus of nalbuphine 10 mg was intravenously injected as a loading dose.PCIA pump solution contained 110 mg nalbuphine diluted to 200 ml with normal saline.The pump was set up with a background infusion at a rate of 4 ml/h, 2 ml bolus dose and 15 min lockout interval.The VAS scores for pain at rest and during activity and uterine contraction pain, Ramsay sedation scores and adverse reactions were observed within 48 h after surgery.Results:The VAS scores for pain at rest and during activity and uterine contraction pain were all ≤3 points, the Ramsay scores were maintained at 2-4 points, hemodynamic parameters were maintained in the normal range, and no adverse reactions such as nausea and vomiting, drowsiness, hyperhidrosis, dizziness, pruritus, and respiratory depression occurred.Conclusion:PCIA with nalbuphine given according to the method mentioned above has good feasibility when used for analgesia following cesarean section.
9.The characteristics of the results of coronary artery CT angiography and coronary artery angiography in 36 patients with probable positive results of treadmill exercise test
Yajun SHI ; Hao WANG ; Peng SHAO ; Hao QIN ; Xilie LU ; Jing JING ; Dongling WEN ; Lichao ZHAO
Chinese Journal of Postgraduates of Medicine 2009;32(z1):8-9
Objective To investigate the clinical characteristics of the results of coronary artery CT angiography and coronary artery angiography in 36 patients with probable positive results of treadmill exercise test(TET).Methods Thirty-six patients due to chest pain received 1ET and coronary artery CT angiography and coronary artery angiography in 2 weeks after admission.The coronary plaques were categorized as noncalcified and calcified plaque on CTA images.Results In 36 patients with probable positive results of TET.23 patients were confirmed as coronary artery disease by coronary artery angiography,and the other 13patients had normal coronary artery.Of 23 coronary artery disease patients,9 patients were presented as onevessel disease,5 patients were presented as two-vessel disease,and 9 patients were presented as three-vessel disease,29 non-calcified plaques and 23 calcified plaques were demeted On CTA.Conclusion TET was useful in confirming the diagnosis of coronary artery disease.The characteristics of the coronary artery plaque is correlated with probable positive results of TET.


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