1.A new method for flow cytometry-based detection of ABO antigen expression levels
Yuyu ZHANG ; Xi LIU ; Junhua XIE ; Bin CAO ; Jiewei ZHENG ; Xinyi ZHU ; Zhongying WANG ; Dong XIANG
Chinese Journal of Blood Transfusion 2025;38(5):665-672
Objective: To design and establish a new method for flow cytometry-based detection of commonly observed highly expressed antigens on red blood cells, and to further evaluate the differences and distribution characteristics of antigen expression levels between ABO blood type homozygotes and heterozygotes in healthy individuals. Methods: Residual blood samples after donor blood type identification by Shanghai Blood Center in April 2024 were collected. Among them, samples of 19 homozygous and 19 heterozygous individuals of type A and type B were selected. Then the expression level of ABO antigen on red blood cells were detected using the new method established in this study and the traditional aldehyde fixed red blood cell method. Both methods were tested independently three times and the results were compared. Results: The mean values of the three detection results of the new method was (×10
/RBC): AA homozygous 3.3±0.5, AO heterozygous 2.8±0.3, BB homozygous 3.6±0.3, BO heterozygous 3.1±2.8. The mean values of the three detection results of the aldehyde fixation method were AA homozygous 5.9±0.9, AO heterozygous 5.0±1.4, BB homozygous 3.8±0.6, and BO heterozygous 3.3±0.4. The average antigen distribution of each genotype followed a normal distribution. Comparing the average antigen expression levels of homozygotes and heterozygotes, both methods showed that A/B homozygotes had higher antigen levels than heterozygotes, with AA being 1.17 to 1.18 times that of AO and BB being 1.15 to 1.16 times that of BO. Comparing the inter batch differences in the three test results of two methods, the new method showed no significant difference in the three test results for four genotypes (P>0.05). The aldehyde fixation method showed significant differences in the test results for all three genotypes (P<0.01) except for BB homozygotes (P>0.05). The reliability and reproducibility of the new method were better than those of the traditional aldehyde fixation method. Conclusion: The antigen expression level of ABO homozygotes is higher than that of heterozygotes, and the difference in antigen level between type A homozygotes and heterozygotes is slightly higher than that of type B. The new method is superior to traditional aldolization fixation methods.
2.Astragalin Regulates Autophagy and Apoptosis of Astrocytes in L4-5 Spinal Dorsal Horn of Mouse Inflammatory Pain Model
Weishan ZHANG ; Jiahong LIN ; Can WANG ; Runheng ZHANG ; Junhua YANG ; Jing LIU ; Guoying LI ; Yuxin MA
Journal of Sun Yat-sen University(Medical Sciences) 2025;46(2):186-196
ObjectiveTo explore the effects of astragalin (AST) on autophagy and apoptosis of astrocytes in the L4-5 dorsal horn of the spinal cord in mice with inflammatory pain induced by complete Freund's adjuvant (CFA). MethodsTwenty-four male C57BL/6 mice, aged six months, were randomly assigned to four groups: control group, saline group, CFA model group, and CFA+AST group, six mice in each group. The inflammatory pain model was established by injection of 10 µL CFA into the right lateral malleolus fossa. The saline group were injected with an equal amount of normal saline at the same site. The inflammatory pain mice in CFA+AST group were further treated with AST (60 mg/kg) intraperitoneally once a day for 21 consecutive days. Multiplex immunofluorescence staining was used to detect the coexpression of autophagy-related factors including ATG 12 and Beclin-1, apoptosis-related factors including Cleaved-Caspase3 and Caspase9, and the astrocyte marker such as GFAP in the L4-5 spinal dorsal horn of the mice in each group. Western blot was used to examine the protein expression levels of autophagy-related proteins(ATG12, Beclin-1) and apoptosis-related proteins(Caspase 3, Caspase 9) in the L4-5 spinal dorsal horn of mice. ResultsImmunofluorescent staining showed that in the L4-5 dorsal horn of the spinal cord, the fluorescence intensity of ATG12 (P<0.000 1) and Beclin-1 (P<0.000 1) was significantly increased, while that of Cleaved-Caspase 3 (P<0.001) and Caspase 9 (P<0.000 1) was decreased in the CFA+AST group when compared to the CFA model group. Furthermore, AST could inhibit the activation of astrocytes. Western blot further confirmed that AST significantly upregulated the expression of ATG12 (P<0.000 1) and Beclin-1 (P<0.000 1) in the L4-5 spinal cord of CFA mice, and downregulated the expression of Caspase 3 (P<0.01) and Caspase 9 (P<0.001). ConclusionsAST promotes autophagy of astrocytes and inhibits their apoptosis in the L4-5 spinal dorsal horn of CFA mice.
3.Dynamic continuous emotion recognition method based on electroencephalography and eye movement signals.
Yangmeng ZOU ; Lilin JIE ; Mingxun WANG ; Yong LIU ; Junhua LI
Journal of Biomedical Engineering 2025;42(1):32-41
Existing emotion recognition research is typically limited to static laboratory settings and has not fully handle the changes in emotional states in dynamic scenarios. To address this problem, this paper proposes a method for dynamic continuous emotion recognition based on electroencephalography (EEG) and eye movement signals. Firstly, an experimental paradigm was designed to cover six dynamic emotion transition scenarios including happy to calm, calm to happy, sad to calm, calm to sad, nervous to calm, and calm to nervous. EEG and eye movement data were collected simultaneously from 20 subjects to fill the gap in current multimodal dynamic continuous emotion datasets. In the valence-arousal two-dimensional space, emotion ratings for stimulus videos were performed every five seconds on a scale of 1 to 9, and dynamic continuous emotion labels were normalized. Subsequently, frequency band features were extracted from the preprocessed EEG and eye movement data. A cascade feature fusion approach was used to effectively combine EEG and eye movement features, generating an information-rich multimodal feature vector. This feature vector was input into four regression models including support vector regression with radial basis function kernel, decision tree, random forest, and K-nearest neighbors, to develop the dynamic continuous emotion recognition model. The results showed that the proposed method achieved the lowest mean square error for valence and arousal across the six dynamic continuous emotions. This approach can accurately recognize various emotion transitions in dynamic situations, offering higher accuracy and robustness compared to using either EEG or eye movement signals alone, making it well-suited for practical applications.
Humans
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Electroencephalography/methods*
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Emotions/physiology*
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Eye Movements/physiology*
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Signal Processing, Computer-Assisted
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Support Vector Machine
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Algorithms
4.Analysis of national external quality assessment results for transfusion compatibility test, 2018 to 2023
Junhua HU ; Peng ZHANG ; Jiali LIU ; Zhiguo WANG ; Yanming LIU ; Shengchen TIAN ; Wanru MA ; Xiang LI ; Xuebin ZHAO ; Feng XUE ; Yuntian WANG ; Dong LIN ; Zheng SUN ; Jiwu GONG ; Lin ZHOU
Chinese Journal of Blood Transfusion 2025;38(12):1720-1727
Objective: To analyze the results of national external quality assessment (EQA) for transfusion compatibility test from 2018 to 2023, with the aim of providing references for improving laboratory testing quality and ensuring the safety of clinical blood transfusion. Methods: Three EQA programs were conducted annually, each distributing 22 quality assessment samples. Participating transfusion laboratories were required to complete testing within specified deadlines and to submit results along with documentation of testing methodologies, reagents, and equipment used. National Center for Clinical Laboratories (NCCL) conducted statistical analysis of laboratory results, evaluated testing outcomes and related circumstances, and provided feedback to participating laboratories. EQA data from transfusion laboratories across China from 2018 to 2023 were collected and systematically analyzed. Results: From 2018 to 2023, the qualification rates for all five items (ABO forward typing, ABO reverse typing, Rh blood group typing, antibody screening, and cross-matching) were 67.59%, 77.11%, 77.38%, 72.78%, 79.96%, and 85.16%, respectively. The mean qualification rates for ABO forward typing, ABO reverse typing, RhD blood group typing, antibody screening, and cross-matching over the past six years were 96.25%±0.59%, 90.45%±4.52%, 96.05%±0.71%, 90.88%±2.86%, and 88.34%±3.48%, respectively. The qualification rates in 2019, 2020, 2022, and 2023 all showed a stable trend of "blood stations>tertiary hospitals>secondary hospitals". The mean qualification rate of laboratories in secondary hospitals from 2018 to 2023 was significantly lower than those of laboratories in tertiary hospitals and blood stations (P<0.05), while no significant difference was observed between laboratories in tertiary hospitals and blood stations (P>0.05). The micro column agglutination method was the most widely used in all five tests. In the four test items, namely ABO forward typing, ABO reverse typing, antibody screening, and cross-matching, there was a statistically significant difference in the qualification rate of micro column agglutination method compared to other methods (P<0.05). There was a statistical difference in the qualification rate between manual and automated detection using micro column agglutination method in the cross-matching tests (P<0.05), whereas no significant difference was noted for the other test items (P>0.05). Conclusion: From 2018 to 2023, the number of laboratories participating in EQA activities has been increasing year by year, and the qualification rate has shown an overall upward trend. The type of laboratory is a key factor affecting the qualification rate, and the testing capabilities of some laboratories still need to be improved. The micro column agglutination method is widely used in transfusion compatibility tests. The established EQA program effectively monitors quality issues in laboratories, drives continuous improvement, and ensures sustained enhancement of testing standards to safeguard clinical blood safety.
5.CT features of abnormally whole-course wide eustachian tubes with microtia and atresia
Junhua LIU ; Fang ZHANG ; Yan SHA
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(2):152-157
Objective:To study the clinical and CT features of the abnormal whole-course wide of eustachian tube (AWWET) with microtia and atresia(MA).Methods:The clinical and CT data of 19 patients (20 ears) from January 2017 to December 2021 with AWWET with MA were retrospectively analyzed, including 15 males and 4 females. The age ranged from 5 to 16 years, with an average of 9.5 years. 50 patients with common MA without wide eustachian tube(ET) as a case control group, including 32 males and 18 females.The age ranged from 5 to 16 years, with an average of 9.2 years. 20 patients (40 ears) who had normal ear CT for tinnitus, otalgia as a normal control group, including 12 males and 8 females. The age ranged from 5 to 16 years, with an average of 12.5 years. We measured the dimension and length of the bony portion of the ET, the total length, the angle between the bony portion and the cartilage portion, and the horizontal angle of ET on CT imagings, and compared with 40 normal ears by SPSS 27.0 software.Results:According to the relationship between AWWET and tympanum, patients were divided into the communicated group and the blocked group. A male predominance, left ear predominance, with high incidence of hemifacial microsomia exhibited in both groups. AWWET was presented as a widened lumen on CT. In 11 ears (4 ears in the communicated group, 7 ears in the blocked group), ETs bifurcated, the upper bony tube extended to the sphenoid body, the lower part continued down to cartilaginous ET and opened onto the nasopharynx, with"mastoid-like"pneumatization of the sphenoid body in 6 ears. The middle ear deformity in case group was more serious than MA control group, especially the blocked group. The incidence of otitis media in the communicated group was lower than that in the MA control group, and 4 cases in the blocked group had effusion in the ET. Compaired with normal ear, the bony ET elongated significantly in the AWWET groups, and the whole course of ET was significantly shortened, specially in the blocked group. The angle between the bony ET and the cartilaginous ET was decreased and the horizontal angle of the ET increased in the AWWET groups, the difference was considered to indicate statistical significance( P<0.05). Conclusions:AWWET with MA is rare, a male predominance, left ear predominance, and with high incidence of hemifacial microsomia. The middle ear deformity is more serious than common MA, especially in the blocked group. The incidence of otitis media in the communicated group is significantly lower than that in the common MA, and the blocked group may be accompanied by ET inflammation.
6.Establishment and application of measurement range of main blood quality indicators in provincial blood stations
Zixuan ZHANG ; Ying CHANG ; Xiaotong ZHANG ; Qingming WANG ; Yuan ZHANG ; Yue LIU ; Qinghua TIAN ; Ka LI ; Guorong LI ; Lixia CHEN ; Junhua SUN ; Yu KANG ; Pingchen HAN ; Xinyu ZHAO ; Song LI
Chinese Journal of Blood Transfusion 2024;37(8):918-926
Objective To obtain the monitoring measurement range of quality indicators of red blood cells,plasma and derivatives and leukocyte-reduced apheresis platelets provided by blood stations in Hebei province,explore the distribution of monitoring values and the change of monitoring level,so as to further strengthen the homogenization construction of quality control laboratories in blood stations in Hebei.Methods In 2023,the sampling data of 12 blood stations in Hebei from 2015 to 2022 were collected,scatter plots were made and the range markers were set,and the"mean±SD"line was taken as the upper limit and lower limit of the measurement range.In 2024,the monitoring values in 2023 were added,and the changes of two measurement ranges were compared to analyze the stability and overall level.Results Comparison of the measurement range from 2015 to 2022 and the measurement range from 2015 to 2023 showed that the standard deviation of the content of deleukocyte suspension of red blood cells-hemoglobin,washed erythrocyte-hemoglobin,washed erythrocyte-su-pernatant protein,cryoprecipitate coagulation factor-FⅧ,fresh frozen plasma-FⅧ,leukocyte-reduced apheresis platelets-leukocyte residue and leukocyte-reduced apheresis platelet-red blood cell concentration decreased from 8.132 to 7.993,6.252 to 6.104,0.273 to 0.267,57.506 to 56.276,0.920 to 0.892,0.653 to 0.644 and 2.653 to 2.603,respectively.The narrowing of the standard deviation range of the above items led to more concentrated monitoring values and reduced disper-sion.Comparison of the measurement range from 2015 to 2022 and the measurement range from 2015 to 2023 showed that the mean value of leukocyte residue of the deleukocyte suspension of red blood cells,hemoglobin content of the wash eryth-rocyte,protein content of supernatant of the wash erythrocyte,hemolysis rate of the wash erythrocyte,FⅧ content of the cryoprecipitate coagulation factor,plasma protein content of the fresh frozen plasma,FⅧ content of the fresh frozen plasma,platelet content of the leukocyte-reduced apheresis platelets changed from 0.362 to 0.476,44.915 to 44.861,0.280 to 0.283,0.137 to 0.142,133.989 to 133.271,60.262 to 60.208,1.301 to 1.277 and 3.036 to 3.033,respectively,and was closer to the national standard line,which reflects an increase in the number of unqualified monitoring values or values close to the national standard line in 2023.The long-term qualified rate of coagulation items was low,and no improvement has been ob-served.The stability of biochemical items has been enhanced but overall deviation has occurred,with the average value close to the national standard line.The possibility of subsequent testing failure has increased.The counting items showed no obvi-ous common characteristics.Conclusion The use of"mean±SD"in the analysis can visually display the distribution of mo-nitoring values of different items in Hebei,forming an indicator measurement range covering the past nine years.It shows the characteristics of each item,and provides reference for subsequent quality control laboratory data analysis of each blood sta-tions to takes active measures to improve the monitoring level.
7.Establishment of the fingerprints of soaking Euodiae fructus with water decoction of Radix glycyrrhizae by Zhangbang method and content determination of alkaloid
Junhua JIANG ; Jianlan CHEN ; Yan XU ; Xuping LIU
China Pharmacist 2024;27(2):220-226
Objective To establish HPLC fingerprints of soaking Euodiae fructus with water decoction of Radix glycyrrhizae by Zhangbang method(hereinafter referred to as"soaking Euodiae fructus"),and to determine the content of evodiamine,rutaecarpine and evocarpine,so as to provide the basis for the quality control and standard improvement of soaking Euodiae fructus.Methods The fingerprint of soaking Euodiae fructus was established based on wavelength switching technology,and the similarity evaluation was conducted.Taking the rutaecarpine as the internal reference,the relative correction factors of evodiamine and evocarpine were calculated by slope correction method,and the differences in measurement results between quantitative analysis of multi-components by single-marker(QAMS)method and external standard method were compared.Results The established fingerprint of soaking Euodiae fructus had a total of 20 common peaks,the similarity between it and the control fingerprint spectrum was 0.970 to 0.998,and 7 common peaks of them were identified.The contents of evodiamine and evocarpine determined by QAMS method were 1.754-7.542 mg/g and 1.281-2.455 mg/g in 10 batches of samples,and the results obtained by QAMS method and external standard method were similar.Conclusion The established HPLC method is simple,reliable,with good separation and repeatability,and can be used for quality evaluation of processed Euodiae fructus.
8.Effect of montelukast combined with budesonide in the treatment of children with intermittent asthma
Gang CHEN ; Jing CAO ; Yurong GENG ; Qian LIU ; Xiaoli LIU ; Junhua SONG
Chinese Journal of Postgraduates of Medicine 2024;47(1):33-38
Objective:To analyze the effect of montelukast combined with budesonide in the treatment of children with intermittent asthma, and the impact on airway remodeling and T helper type 1 (Th1)/T helper type 2 (Th1/Th2) related cytokines.Methods:A prospective study was conducted among 120 children with intermittent asthma admitted to Huanghua Municipal People′s Hospital from December 2021 to February 2023. The children were randomly divided into the control group (60 children treated with budesonide atomizationinhalation) and the observation group (60 children treated with montelukast on the basis of the treatment of control group). Clinical efficacy, airway remodeling indicators [total area of airway (Ao), outer diameter of airway (D) and wall area to total airway cross-sectional area (WA%)], pulmonary function [peak expiratory flow (PEF), forced expiratory volume in 1 second (FEV 1)/forced vital capacity (FVC) and the maximum expiratory flow at 25% of vital capacity (MEF25%)], Th1/Th2 related cytokines, inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), interleukin-6 (IL-6) and interferon-gamma (IFN-γ)], recurrence, and the incidence of adverse reactions were compared between the two groups. Results:The total effective rate in the observation group was higher than that in the control group: 90.00% (54/60) vs. 75.00% (45/60) ( P<0.05). After treatment, Ao, D and WA% in the observation group were lower than those in the control group: (17.58 ± 1.89) mm 2 vs. (19.22 ± 1.94) mm 2, (4.25 ± 0.48) mm vs. (4.48 ± 0.49) mm, (63.75 ± 6.49)% vs. (69.22 ± 7.14)% ( P<0.05). PEF, FEV 1/FVC and MEF25% in the observation group were higher than those in the control group: (3.13 ± 0.34) L/s vs. (2.86 ± 0.35) L/s, (87.45 ± 8.86) % vs. (83.59 ± 8.42) %, (87.63 ± 8.86)% vs. (82.15 ± 8.43)% ( P<0.05). The levels of Th1 and Th1/Th2 in the observation group were higher than those in the control group: (14.13 ± 1.46) % vs. (10.27 ± 1.25) %, 3.46 ± 0.39 vs. 1.88 ± 0.25, and the level of Th2 was lower than that in the control group: (3.96 ± 0.45)% vs. (5.48 ± 0.56)% ( P<0.05). After treatment, the levels of TNF-α and IFN-γ in the observation group were higher than those in the control group: (76.15 ± 7.78) ng/L vs. (66.38 ± 6.47) ng/L, (7.15 ± 0.74) ng/L vs. (6.14 ± 0.66) ng/L. The levels of IL-4 and IL-6 were lower than those in the control group: (77.85 ± 7.96) ng/L vs. (86.42 ± 8.74) ng/L, (37.25 ± 3.89) mg/L vs. (44.23 ± 4.57) mg/L ( P<0.05). The recurrence rate in the observation group was lower than that in the control group: 3.33% (2/60) vs. 15.00% (9/60) ( P<0.05). The incidence rates of adverse reactions in the two groups were without statistically significant difference between the groups ( P>0.05). Conclusions:Montelukast combined with budesonide can reduce airway remodeling in children with intermittent asthma, improve their pulmonary function, Th1/Th2 related cytokines and inflammatory response indicators, and reduce recurrence rate, with good safety.
9.Licorice-saponin A3 is a broad-spectrum inhibitor for COVID-19 by targeting viral spike and anti-inflammation
Yang YI ; Wenzhe LI ; Kefang LIU ; Heng XUE ; Rong YU ; Meng ZHANG ; Yang-Oujie BAO ; Xinyuan LAI ; Jingjing FAN ; Yuxi HUANG ; Jing WANG ; Xiaomeng SHI ; Junhua LI ; Hongping WEI ; Kuanhui XIANG ; Linjie LI ; Rong ZHANG ; Xin ZHAO ; Xue QIAO ; Hang YANG ; Min YE
Journal of Pharmaceutical Analysis 2024;14(1):115-127
Currently,human health due to corona virus disease 2019(COVID-19)pandemic has been seriously threatened.The coronavirus severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike(S)protein plays a crucial role in virus transmission and several S-based therapeutic approaches have been approved for the treatment of COVID-19.However,the efficacy is compromised by the SARS-CoV-2 evolvement and mutation.Here we report the SARS-CoV-2 S protein receptor-binding domain(RBD)inhibitor licorice-saponin A3(A3)could widely inhibit RBD of SARS-CoV-2 variants,including Beta,Delta,and Omicron BA.1,XBB and BQ1.1.Furthermore,A3 could potently inhibit SARS-CoV-2 Omicron virus in Vero E6 cells,with EC50 of 1.016 pM.The mechanism was related to binding with Y453 of RBD deter-mined by hydrogen-deuterium exchange mass spectrometry(HDX-MS)analysis combined with quan-tum mechanics/molecular mechanics(QM/MM)simulations.Interestingly,phosphoproteomics analysis and multi fluorescent immunohistochemistry(mIHC)respectively indicated that A3 also inhibits host inflammation by directly modulating the JNK and p38 mitogen-activated protein kinase(MAPK)path-ways and rebalancing the corresponding immune dysregulation.This work supports A3 as a promising broad-spectrum small molecule drug candidate for COVID-19.
10.Efficacy and safety of simultaneous integrated boost intensity-modulated radiotherapy for the treatment of lung cancer brain metastases
Jing YANG ; Fang LIU ; Meili MA ; Dongshuai HUANG ; Junhua WANG
Journal of International Oncology 2024;51(10):609-613
Objective:To analyze the therapeutic effect and safety of simultaneous integrated boost intensity-modulated radiotherapy for lung cancer brain metastases.Methods:A total of 300 patients with lung cancer brain metastases admitted to the Department of Oncology, Qingdao Municipal Hospital from March 2021 to March 2023 were selected as the study objects. The patients were divided into control group ( n=150) and study group ( n=150) by random number table method. The control group received sequential three-dimensional conformal radiotherapy, while the study group received simultaneous integrated boost intensity-modulated radiotherapy. The short-term and medium-term efficacy, target dose, and adverse reactions were compared between the two groups. Results:The short-term and medium-term total effective rates of the study group were 73.33% (110/150) and 88.67% (133/150), respectively, which were higher than those of the control group [51.33% (77/150) and 71.33% (107/150) ] ( χ2=15.46, P<0.001; χ2=14.08, P<0.001). The D min in gross tumor planning target volume and whole brain clinical planning target volume of the study group were (23.78±1.11) and (58.46±0.55) Gy, respectively, which were higher than those in the control group [ (16.67±1.08) and (53.44±0.74) Gy], with statistically significant differences ( t=56.22, P<0.001; t=66.68, P<0.001). The D mean in gross tumor planning target volume and whole brain clinical planning target volume of the study group were (44.12±0.87) and (62.19±0.57) Gy, respectively, which were higher than those in the control group [ (37.55±0.89) and (57.78±0.82) Gy], with statistically significant differences ( t=64.65, P<0.001; t=54.08, P<0.001). The total incidence of adverse reactions was 30.67% (46/150) in the study group and 36.67% (55/150) in the control group, with no significantly significant difference ( χ2=1.20, P=0.271) . Conclusion:Compared with sequential three-dimensional conformal radiotherapy, simultaneous integrated boost intensity-modulated radiotherapy has better short-term and medium-term efficacy in patients with lung cancer brain metastases. Target dose can be increased without increasing adverse reactions.

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