1.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.
2.Pathological diagnosis of solid pancreatic lesions with endoscopic ultrasound-guided fine needle aspiration: a series study of 311 cases
Xiaoxiao WEN ; Xiaoyi LIU ; Jinfeng CUI ; Lichao ZHANG ; Wenxuan LIU ; Haiyan YANG ; Yuan WANG ; Li YI ; Lei LOU ; Juan WANG ; Yuehong LI ; Wenxin WU ; Xianghong ZHANG
Chinese Journal of Pathology 2025;54(1):52-58
Objective:To investigate the combined application of cytology, cell block histology and immunohistochemistry to improve the diagnostic accuracy of solid pancreatic lesions in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) samples.Methods:The pathological data of EUS-FNA in 311 cases of solid pancreatic lesions submitted to the Second Hospital of Hebei Medical University, Shijiazhuang, China from May 2019 to September 2023 were retrospectively analyzed. The cases included pancreatic ductal adenocarcinoma (PDAC, 172 cases), solid pseudopapillary neoplasm (SPN, 12 cases), neuroendocrine tumors (PNET, 14 cases) and chronic pancreatitis (113 cases). The cytological features of smears, the histology of cell block sections and the diagnostic markers in PDAC, SPN and PNET were analyzed. The diagnostic accuracies of cytology, cell block histology/immunohistochemistry and combination of the two methods for classifying these pancreatic solid lesions were evaluated.Results:Irregular arrangement of atypical (cancer) cells, anisonucleosis and nuclear atypia were the typical cytological features of PDAC, while presence of pseudopapillae with a myxoid/hyalinized fibrovascular core and low adhesion/salt-and-pepper chromatin were diagnostic features of SPN and NET, respectively. Immunohistochemical results showed that CK7 and CK19 were the most sensitive markers of pancreatic ductal epithelia, and the diffuse strong expression of S-100P (102/111, 91.9%) and aberrant expression of p53 (80/111, 72.1%) were important immunophenotypic markers of PDAC. Various degrees of CDX2 expression could be found in 66.4% PDAC. The expression of CD10, PR, vimentin, CD99 and cyclinD1 and the aberrant expression of β-catenin were the immunophenotypic features of SPN, while the expression of CgA, Syn and CD56 were indispensable immunemarkers for the diagnosis of PNET. Overall, cytology had higher sensitivity than cell block histology (93.9% versus 82.8%) and lower specificity (92.9% versus 99.1%), while the combination of the two methods significantly improved the sensitivity to 96.9% in solid pancreatic lesions. The combination of cytology and cell block histology could significantly improve the diagnostic efficacy of EUS-FNA in PDAC.Conclusions:Integrated diagnosis based on cytology (including rapid on-site evaluation), cell block histology and immunohistochemical findings could significantly improve the diagnostic yield of EUS-FNA in classifying solid pancreatic lesions.
3.Pathological diagnosis of solid pancreatic lesions with endoscopic ultrasound-guided fine needle aspiration: a series study of 311 cases
Xiaoxiao WEN ; Xiaoyi LIU ; Jinfeng CUI ; Lichao ZHANG ; Wenxuan LIU ; Haiyan YANG ; Yuan WANG ; Li YI ; Lei LOU ; Juan WANG ; Yuehong LI ; Wenxin WU ; Xianghong ZHANG
Chinese Journal of Pathology 2025;54(1):52-58
Objective:To investigate the combined application of cytology, cell block histology and immunohistochemistry to improve the diagnostic accuracy of solid pancreatic lesions in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) samples.Methods:The pathological data of EUS-FNA in 311 cases of solid pancreatic lesions submitted to the Second Hospital of Hebei Medical University, Shijiazhuang, China from May 2019 to September 2023 were retrospectively analyzed. The cases included pancreatic ductal adenocarcinoma (PDAC, 172 cases), solid pseudopapillary neoplasm (SPN, 12 cases), neuroendocrine tumors (PNET, 14 cases) and chronic pancreatitis (113 cases). The cytological features of smears, the histology of cell block sections and the diagnostic markers in PDAC, SPN and PNET were analyzed. The diagnostic accuracies of cytology, cell block histology/immunohistochemistry and combination of the two methods for classifying these pancreatic solid lesions were evaluated.Results:Irregular arrangement of atypical (cancer) cells, anisonucleosis and nuclear atypia were the typical cytological features of PDAC, while presence of pseudopapillae with a myxoid/hyalinized fibrovascular core and low adhesion/salt-and-pepper chromatin were diagnostic features of SPN and NET, respectively. Immunohistochemical results showed that CK7 and CK19 were the most sensitive markers of pancreatic ductal epithelia, and the diffuse strong expression of S-100P (102/111, 91.9%) and aberrant expression of p53 (80/111, 72.1%) were important immunophenotypic markers of PDAC. Various degrees of CDX2 expression could be found in 66.4% PDAC. The expression of CD10, PR, vimentin, CD99 and cyclinD1 and the aberrant expression of β-catenin were the immunophenotypic features of SPN, while the expression of CgA, Syn and CD56 were indispensable immunemarkers for the diagnosis of PNET. Overall, cytology had higher sensitivity than cell block histology (93.9% versus 82.8%) and lower specificity (92.9% versus 99.1%), while the combination of the two methods significantly improved the sensitivity to 96.9% in solid pancreatic lesions. The combination of cytology and cell block histology could significantly improve the diagnostic efficacy of EUS-FNA in PDAC.Conclusions:Integrated diagnosis based on cytology (including rapid on-site evaluation), cell block histology and immunohistochemical findings could significantly improve the diagnostic yield of EUS-FNA in classifying solid pancreatic lesions.
4.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.
5.Research on phase modulation to enhance the feature of high-frequency steady-state asymmetric visual evoked potentials.
Wei ZHAO ; Lichao XU ; Xiaolin XIAO ; Weibo YI ; Yuanfang CHEN ; Kun WANG ; Minpeng XU ; Dong MING
Journal of Biomedical Engineering 2023;40(3):409-417
High-frequency steady-state asymmetric visual evoked potential (SSaVEP) provides a new paradigm for designing comfortable and practical brain-computer interface (BCI) systems. However, due to the weak amplitude and strong noise of high-frequency signals, it is of great significance to study how to enhance their signal features. In this study, a 30 Hz high-frequency visual stimulus was used, and the peripheral visual field was equally divided into eight annular sectors. Eight kinds of annular sector pairs were selected based on the mapping relationship of visual space onto the primary visual cortex (V1), and three phases (in-phase[0º, 0º], anti-phase [0º, 180º], and anti-phase [180º, 0º]) were designed for each annular sector pair to explore response intensity and signal-to-noise ratio under phase modulation. A total of 8 healthy subjects were recruited in the experiment. The results showed that three annular sector pairs exhibited significant differences in SSaVEP features under phase modulation at 30 Hz high-frequency stimulation. And the spatial feature analysis showed that the two types of features of the annular sector pair in the lower visual field were significantly higher than those in the upper visual field. This study further used the filter bank and ensemble task-related component analysis to calculate the classification accuracy of annular sector pairs under three-phase modulations, and the average accuracy was up to 91.5%, which proved that the phase-modulated SSaVEP features could be used to encode high- frequency SSaVEP. In summary, the results of this study provide new ideas for enhancing the features of high-frequency SSaVEP signals and expanding the instruction set of the traditional steady state visual evoked potential paradigm.
Humans
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Evoked Potentials, Visual
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Brain-Computer Interfaces
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Healthy Volunteers
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Signal-To-Noise Ratio
6.Progress of classification algorithms for motor imagery electroencephalogram signals.
Tuo LIU ; Yangyang YE ; Kun WANG ; Lichao XU ; Weibo YI ; Minpeng XU ; Dong MING
Journal of Biomedical Engineering 2021;38(5):995-1002
Motor imagery (MI), motion intention of the specific body without actual movements, has attracted wide attention in fields as neuroscience. Classification algorithms for motor imagery electroencephalogram (MI-EEG) signals are able to distinguish different MI tasks based on the physiological information contained by the EEG signals, especially the features extracted from them. In recent years, there have been some new advances in classification algorithms for MI-EEG signals in terms of classifiers versus machine learning strategies. In terms of classifiers, traditional machine learning classifiers have been improved by some researchers, deep learning and Riemannian geometry classifiers have been widely applied as well. In terms of machine learning strategies, ensemble learning, adaptive learning, and transfer learning strategies have been utilized to improve classification accuracies or reach other targets. This paper reviewed the progress of classification algorithms for MI-EEG signals, summarized and evaluated the existing classifiers and machine learning strategies, to provide new ideas for developing classification algorithms with higher performance.
Algorithms
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Brain-Computer Interfaces
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Electroencephalography
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Imagery, Psychotherapy
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Imagination
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Machine Learning
7.Effects of perindopril on bone metabolism in a rat model of retinoic acid-induced osteoporosis
Yi ZHONG ; Qing XUE ; Yi ZHOU ; Lianghua CHEN ; Lichao ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(18):2589-2595
BACKGROUND: Renin-angiotensin-aldosterone system existed in bone tissue. Recent studies on antihypertensive drugs found that angiotensin converting enzyme inhibitor type antihypertensive drug was possibly effective for osteoporosis. Perindopril is one of the commonly used antihypertensive drugs. Whether perindopril affected bone metabolism or could be used in anti-osteoporosis has not been reported.
OBJECTIVE: To observe effects of perindopril on bone metabolism in a rat model of osteoporosis induced by retinoic acid.
METHODS: Fifty Sprague-Dawley rats were randomly divided into five groups, with ten in each group. In the model group and each perindopril groups, rats were intragastricaly administered retinoic acid solution 80 mg/kg per day. After successful model establishment, rats in different perindopril groups were intragastrical y administered perindopril 2, 4 and 8 mg/kg per day, once a day, for 42 consecutive days. In the normal control and model groups, rats were given an equal volume of distil ed water. Serum calcium, phosphorus, alkaline phosphatase, bone mass and bone mineral density were detected in each group. Expression of bone specific alkaline phosphatase and tartrate-resistant acid phosphatase mRNA in bone tissue was determined.
RESULTS AND CONCLUSION: Compared with the model group, after treatment with perindopril, serum calcium and phosphorus levels were increased, alkaline phosphatase activities were significantly decreased, bone mass and bone mineral density were obviously increased in rats with retinoic acid-induced osteoporosis. Expression of bone specific alkaline phosphatase mRNA was higher in the perindopril 8 mg/kg group than in the perindopril 2 and 4 mg/kg groups and model group. Tartrate-resistant acid phosphatase mRNA expression was higher in the perindopril 8 mg/kg group than in the model group. These results indicated that perindopril could improve partial bone metabolic biochemical markers in osteoporosis rats, promoted bone formation by up-regulating bone specific alkaline phosphatase mRNA expression, and had a certain preventive effect on retinoic acid-induced osteoporosis.
8.Evaluation of efficacy and safety of a focal fractional laser for the treatment of atrophic acne scars
Hongjin WU ; Bingrong ZHOU ; Shufen XIE ; Jia′an ZHANG ; Jin LI ; Juan LIU ; Fei YI ; Shen WANG ; Lichao ZHANG ; Dan LUO
Chinese Journal of Dermatology 2015;(12):881-885
Objective To evaluate the efficacy and safety of focal fractional laser treatment(FFLT)for atrophic acne scars. Methods A randomized, self-controlled study was performed. A total of 20 patients with atrophic facial acne scars were enrolled into this study. Treatments were randomly administered in a split-face manner. Half of each subject′s face received FFLT(FFLT side), and the other half underwent full-face fractional CO2 laser resurfacing(control side), for one session. All the patients were followed up for 3 months after the treatment. Evaluation was based on the ECCA grading scale (échelle d′évaluation clinique des cicatrices d′acné)and patient satisfaction score. A VISIA skin detector was used to take photographs and evaluate skin texture. Moreover, physical parameters of the skin, including erythema index, melanin index and transepidermal water loss (TEWL), were measured. Adverse effects were recorded and evaluated. Statistical analysis was carried out by paired t test, Wilcoxon paired rank test, Fisher′s exact test and repeated-measure analysis of variance. Results The ECCA score decreased from 51.24 ± 17.61 at the baseline to 34.46 ± 14.99 at 3 months after the treatment at the FFLT side(t = 7.886, P < 0.05), and from 50.96 ± 18.96 to 38.29 ± 14.86 at the control side(t =6.123, P < 0.05), and was significantly lower in the FFLT side than in the control side (t = 4.462, P < 0.05)at 3 months after the treatment. The improvement rate was significantly higher in the FFLT side than in the control side (32.75% vs. 24.86%, P = 0.016 by Fisher′s exact test)at 3 months after the treatment. Decreased pain and edema scores were observed at the FFLT side compared with the control side at 1 hour after the treatment (both P < 0.05), but no significant difference was noted in the duration of erythema or crusting between the two sides (both P > 0.05). Compared with those before the treatment, skin texture scores decreased in both sides (both P < 0.05), and were significantly lower in the FFLT side than in the control side at 3 months after the treatment(P < 0.05). The erythema index was significantly lower in the FFLT side than in the control side in both scarred areas and non-scarred areas on day 1 after the treatment (both P < 0.05). Both melanin index and TEWL at the FFLT side were significantly increased in scarred areas, but decreased in non-scarred areas compared with those at the control side within 3 days after the treatment (all P < 0.05). Similarly, the water content of the stratum corneum at the FFLT side was significantly lower in scarred areas, but higher in non-scarred areas compared with that at the control side between day 1 and 7 after the treatment (both P < 0.05). No significant difference was observed in the erythema index, TEWL or water content of the stratum corneum between the FFLT side and control side at scarred areas or non-scarred areas(all P > 0.05)from 2 weeks to 3 months after the treatment(all P > 0.05). Conclusion FFLT can improve therapeutic outcomes in atrophic acne scars with reduced adverse reactions.
9.Combined effects of cytarabine and PDT treatment on leukemia cell HL-60
Yi LIU ; Huijuan YIN ; Lichao GU ; Yingxin LI ; Tianjun LIU
International Journal of Biomedical Engineering 2014;37(4):217-221
Objective To evaluate the combined effects of photodynamic therapy (PDT) with PSD-007 and cytarabine (Ara-c) on human acute promyelocyte leukemia cell HL-60.Methods The experiments were divided into four groups:control group,PDT-only groups (PDT 1-4 groups:the combination of PSD-007 concentrations (5 μg/ml and 7.5 μg/ml) and the energy density of laser (EDL) (1.2 J/cm2 and 2.4 J/cm2),Ara-c-only groups (Ara-c A group:0.3μg/ml,Ara-c B group:1.2μg/ml) and combination groups (the pair-wise combinations of the PDT doses and Ara-c doses above).All combination groups were treated with three treating methods,including P24A (the cells were treated with PDT for 24 h,and then cocultured with Ara-c for another 24 h),A24P (treated with Ara-c for 24 h before PDT,and then cocultured with PDT for 24 h),and PA24 (treated with the Ara-c and PDT for 24 h).CCK-8 method was used to test the cell viability,and the combined effect was analyzed using King formula.The changes of cell cycles were analyzed using flow cytometry.Results In the case of low-dose PDT,the combination groups showed coordinated effect with all three methods,whereas in the case of high-dose PDT,they showed additive or coordinated effect in P24A and A24P,but it showed antagonistic effect in the schedule of PA24.Cell cycles were inhibited to G0/G1 phase by both PDT and Ara-c.Conclusions Coordinated effects could be found when HL-60 cells were treated with the combination of Ara-c and PDT.The effects depended on the dose of Ara-c and PDT and the operation schedules.The effects at low dose were more obvious than that of high dose,and allowing a 24 h period in between the addition of PDT and Ara-c also promoted the effects.

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