1.Construction of a system for isolation and purification of NK cells from whole blood donations
Tengyu CAO ; Huayu LIN ; Xuanzhi ZHANG ; Cuimi DUAN ; Yi LIU ; Xiaonan XUE ; Liping SUN ; Yang YU
Chinese Journal of Blood Transfusion 2025;38(2):181-188
[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×108vs (6.49±2.16)×108, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.
2.Herbal Textual Research on Moschus in Famous Classical Formulas
Juanjuan LIU ; Sini LI ; Jie JI ; Liping YANG ; Houkang CAO ; Xiaohui MA ; Ling JIN ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):194-202
In this paper, by consulting the ancient and modern literature, the name, origin, quality evaluation, harvesting and processing, and others of the original animal and medicinal materials of Moschus were systematically sorted out and verified, in order to provide the basis for the development and utilization of the famous classical formulas containing Moschus. According to the textual research, musk deer was first recorded in Shanhaijing. Shennong Bencaojing was recorded as Moschus and all generations were used as the correct name, but there were also aliases such as Shefu, Xiangzhang and Xiangqizi. In ancient times, Moschus berezovskii, M. sifanicus and M. moschiferus were the main sources of Moschus, and the quality of Moschus produced in northwest China was better than that produced in the Yangtze River basin. In modern times, Moschus of M. moschiferus produced in northeast China, M. sifanicus produced in Gansu, Sichuan and other places, and M. berezovskii produced in Ningxia, Shaanxi and other places are regarded as genuine. In ancient times, gunshots, lassoes, arrow shots and other methods were generally used to hunt live musk deer, and the sachets were immediately cut off. Those with high quality were called Xiangshanhuo, and dried in the shade after harvesting, which was known as Maoke Shexiang. Cut open the sachet, remove the shell and dry preservation, commonly known as Moschus kernel. In modern times, the method of taking Moschus from the living body of cultured musk deer is adopted, that is, Moschus kernel is directly taken from its sachet, dried in the shade or dried in a closed dryer. This method realizes the sustainable utilization of Chinese herbal medicine resources, but attention should be paid to the frequency and quality of Moschus. The harvesting time is mostly after the autumnal equinox every year, and before the next summer, it is better to gather sachet in winter. In recent times, it is believed that the shell Moschus is dry, full, thin, elastic, loose inside, many particles, strong and persistent aroma for the best, while the Moschus kernel is particle purple-black, powder yellow-brown, soft and oily texture, strong and persistent aroma for the best. The ancient processing method of Moschus was extracting kernels from the shell. After removing impurities, it is ground and used as medicine. Because its composition is not suitable for heating, the processing method is most common in preparations such as grinding into powder and putting into pills or powders, which has the effect of opening up the orifices and refreshing the mind, and it has continued to this day. Based on the research conclusions, it is suggested that the development of famous classical formulas containing Moschus, M. sifanicus, M. moschiferus and M. berezovskii should be used as the origins. According to the processing requirements specified in the original formula, it should be processed and used as medicine, while those without processing requirements should be used as raw products.
4.Compound Centella asiatica formula alleviates Schistosoma japonicum-induced liver fibrosis in mice by inhibiting the inflammation-fibrosis cascade via regulating the TLR4/MyD88 pathway.
Liping GUAN ; Yan YAN ; Xinyi LU ; Zhifeng LI ; Hui GAO ; Dong CAO ; Chenxi HOU ; Jingyu ZENG ; Xinyi LI ; Yang ZHAO ; Junjie WANG ; Huilong FANG
Journal of Southern Medical University 2025;45(6):1307-1316
OBJECTIVES:
To explore the therapeutic mechanism of compound Centella asiatica formula (CCA) for alleviating Schistosoma japonicum (Sj)-induced liver fibrosis in mice.
METHODS:
The active components and targets of CCA were identified using the TCMSP database with cross-analysis of Sj-related liver fibrosis targets. A "drug-component-target-pathway-disease" network was constructed using Cytoscape 3.9.1. Functional enrichment analysis (GO/KEGG) was performed using DAVID. Molecular docking study was carried out to validate interactions between the core targets and the key compounds. For experimental validation of the results, 36 mice were divided into control group, Sj-infected model group, and CCA-treated groups. In the latter two groups, liver fibrosis was induced via abdominal infection with Sj cercariae for 8 weeks, followed by 8 weeks of daily treatment with CCA decoction or saline. Hepatic pathology of the mice was assessedwith HE and Masson staining, and hepatic expressions of collagen-I and collagen-III were detected using immunohistochemistry; serum IL-6 and TNF-α levels were determined with ELISA. Hepatic expressions of TLR4 and MyD88 proteins were analyzed with Western blotting.
RESULTS:
We identified a total of 107 bioactive CCA components and 791 targets, including 37 intersection targets linked to Sj-induced fibrosis. The core targets included TNF, TP53, JUN, MMP9, and CXCL8, involving the IL-17 signaling, lipid metabolism, TLR4/MyD88 axis, and cancer pathways. Molecular docking study confirmed strong binding affinity between quercetin (a primary CCA component) and TNF/TP53/JUN/MMP9. In Sj-infected mouse models, CCA treatment significantly attenuated hepatic inflammatory cell infiltration, reduced collagen-I and collagen-III deposition, improved tissue architecture, reduced serum IL-6 and TNF-α levels, and downregulated TLR4 and MyD88 expressions in the liver.
CONCLUSIONS
CCA mitigates Sj-induced liver fibrosis by targeting TNF, TP53, JUN, and MMP9 to modulate the TLR4/MyD88 pathway, thereby suppressing pro-inflammatory cytokine release, inhibiting hepatic stellate cell activation, reducing collagen deposition, and preventing granuloma formation in the liver.
Animals
;
Toll-Like Receptor 4/metabolism*
;
Mice
;
Myeloid Differentiation Factor 88/metabolism*
;
Schistosoma japonicum
;
Liver Cirrhosis/parasitology*
;
Schistosomiasis japonica
;
Signal Transduction
;
Molecular Docking Simulation
;
Inflammation
;
Centella/chemistry*
;
Drugs, Chinese Herbal/pharmacology*
;
Tumor Necrosis Factor-alpha/metabolism*
5.A Method for Detecting Depression in Adolescence Based on an Affective Brain-Computer Interface and Resting-State Electroencephalogram Signals.
Zijing GUAN ; Xiaofei ZHANG ; Weichen HUANG ; Kendi LI ; Di CHEN ; Weiming LI ; Jiaqi SUN ; Lei CHEN ; Yimiao MAO ; Huijun SUN ; Xiongzi TANG ; Liping CAO ; Yuanqing LI
Neuroscience Bulletin 2025;41(3):434-448
Depression is increasingly prevalent among adolescents and can profoundly impact their lives. However, the early detection of depression is often hindered by the time-consuming diagnostic process and the absence of objective biomarkers. In this study, we propose a novel approach for depression detection based on an affective brain-computer interface (aBCI) and the resting-state electroencephalogram (EEG). By fusing EEG features associated with both emotional and resting states, our method captures comprehensive depression-related information. The final depression detection model, derived through decision fusion with multiple independent models, further enhances detection efficacy. Our experiments involved 40 adolescents with depression and 40 matched controls. The proposed model achieved an accuracy of 86.54% on cross-validation and 88.20% on the independent test set, demonstrating the efficiency of multimodal fusion. In addition, further analysis revealed distinct brain activity patterns between the two groups across different modalities. These findings hold promise for new directions in depression detection and intervention.
Humans
;
Male
;
Female
;
Adolescent
;
Case-Control Studies
;
Depression/diagnosis*
;
Early Diagnosis
;
Rest
;
Electroencephalography/methods*
;
Brain-Computer Interfaces
;
Models, Psychological
;
Reproducibility of Results
;
Affect/physiology*
;
Photic Stimulation/methods*
;
Video Recording
;
Brain/physiopathology*
6.Retrospective analysis of unexpected antibodies in primary-screened D-negative blood donors
Hecai YANG ; Yin GUAN ; Xiaoli MA ; Yonglei LYU ; Minglu GENG ; Yi CAO ; Liping WANG ; Hongwei MA
Chinese Journal of Blood Transfusion 2025;38(11):1556-1561
Objective: To analyze the frequency and investigate the causes of unexpected antibodies in D-negative blood donors. Methods: From January 2022 to December 2024, 3 768 D-negative blood donors sent to our laboratory were selected as research subjects. D-negative confirmation test and RhCE phenotype detection were applied by saline tube method and microcolumn gel indirect antiglobulin test (IAT), respectively. Antibody screening and identification were performed using the polybrene method and IAT column agglutination methods. Anti-D, anti-C and anti-G specificities were identified by a two-step adsorption-elution method, and the genotypes of D-negative samples were determined by RHD gene amplification, Sanger sequencing, and PacBio Single Molecule Real-Time (SMRT) sequencing. Results: Among D-negative donors, ccee and Ccee phenotypes accounted for the highest proportion, 55.68% (2 098/3 768) and 29.56% (1 114/3 768), respectively, while CcEE and CCEe phenotypes were the least, with one case detected in each, accounting for 0.03% (1/3 768). A total of 165 cases with D variant phenotype were detected, and the proportion of D variant was 4.38% (165/3 768) in the donors detected by D-negative confirmation test. Antibody screening positive blood donors were identified in 93 cases with a proportion of 2.47% (93/3 768). Antibody specificity was determined in 84 blood donors, and 9 samples showed no clear specificity. Anti-D was detected most frequently (n=68), in which 6 of them were detected having multiple antibodies, anti-D + anti-C (n=2), anti-D + anti-G(n=1), and anti-D + anti-E(n=3). The other antibodies detected were anti-E (n=1), anti-M(n=9), anti-P1 (n=3), anti-Le
(n=1), and anti-HI(n=2). Fourteen cases were detected with anti-D in serological D-negative donors with C+ or E+ phenotype, in which three of them were DVI type 3 individuals and 11 cases were D negative individuals. Conclusion: The incidence of unexpected antibodies was higher in D-negative blood donors than in the total donors, with anti-D being the most common. The data provide insights for prevention and monitoring hemolytic disease of the fetus and newborn (HDFN) caused by anti-D. To ensure the safety of blood transfusion, routine unexpected antibody screening for RhD-negative blood donors is recommended to prevent the use of unexpected antibodies positive plasma in the clinic.
7.Analysis of life expectancy and cause-eliminated life expectancy of the registered residents in Qingpu District, Shanghai, 2002‒2021
Rongrong HAN ; Shan JIN ; Liping FANG ; Wu WANG ; Jieyu ZHANG ; Bo CAO ; Xingxing XU
Shanghai Journal of Preventive Medicine 2024;36(9):878-882
ObjectiveTo analyze the trends in life expectancy and cause-eliminated life expectancy of the registered residents in Qingpu District, Shanghai from 2002 to 2021, and to identify the major diseases causing life expectancy loss in the area, so as to provide a theoretical basis for strengthening disease prevention and interventions and optimizing health resources allocation. MethodsUsing the International Classification of Diseases (ICD)-10 classification of death causes, average life expectancy and cause-eliminated life expectancy were calculated with the abbreviated life table and the cause-eliminated life expectancy table. A trend forecast of life expectancy from 2022 to 2029 was made. ResultsFrom 2002 to 2021, the life expectancy of Qingpu District residents showed an increasing trend, with an average annual growth of 0.32 years (AAPC=0.39%, P<0.001). Among the seven major causes of death, the cause-eliminated expected life expectancy for circulatory system diseases showed the most significant upward trend (AAPC=0.53%, P<0.001), rising from the third leading cause of death in 2002 to the first in 2021. The life expectancy loss rate due to circulatory system diseases increased from 3.79% to 7.97%. Respiratory system diseases showed the largest decline, decreasing from the first leading cause of death in 2002 to the fourth in 2021, with the life expectancy loss rate decreasing from 6.83% to 0.99%. ConclusionLife expectancy of the registered residents in Qingpu District has reached a leading level in China. Future efforts should focus on effectively increasing the life expectancy of male residents, strengthening the comprehensive prevention and treatment of chronic diseases such as cerebrovascular diseases, diabetes, and malignant tumors, and improving health promotion policies for residents to achieve a steady long-term increase in regional life expectancy.
8.Molluscicidal effect of spraying different formulations of niclosamide ethanolamine salt with drones against Oncomelania hupensis in ditches
Zelin ZHU ; Xia ZHANG ; Junyi HE ; Ying CHEN ; Weisi WANG ; Hehua HU ; Chunli CAO ; Ziping BAO ; Suying GUO ; Liping DUAN ; Yi YUAN ; Jing XU ; Shizhu LI ; Xiaonong ZHOU
Chinese Journal of Schistosomiasis Control 2024;36(5):527-530
Objective To evaluate the molluscicidal effect of spraying different formulations of niclosamide ethanolamine salt with drones against Oncomelania hupensis in ditches. Methods A semi-dry and semi-wet ditch with O. hupensis snails was selected in the second branch field of Jiangbei Farm, Jiangling County, Hubei Province in May 2023, and divided into 4 experimental areas, named groups A1, A2, B1 and B2. Environmental cleaning was performed in groups A1 and B2, and was not conducted in groups A2 or B2. Then, 50% wettable powder of niclosamide ethanolamine salt was sprayed with drones at an effective dose of 2 g/m2 in groups A1 and A2, and 5% niclosamide ethanolamine salt granule was sprayed with drones at an effective dose of 2 g/m2 in groups B1 and B2. O. hupensis snails were surveyed using the systematic sampling method 1, 3, 5, 7, 14 days after spraying, and the natural mortality and corrected mortality of O. hupensis snails were calculated. Results The occurrence of frames with living snails, mean density of living snails and natural mortality of snails were 97.50% (117/120), 6.30 snails/0.1 m2 and 1.18% (9/765) in the test ditch before spraying, respectively. There were significant differences in the mortality of snails among four groups 1, 3, 5, 7 and 14 days after spraying niclosamide formulations with drones (χ2 = 17.230, 51.707, 65.184, 204.050 and 34.435, all P values < 0.01). The overall mortality rates of snails were 94.51% (1 051/1 112), 79.44% (908/1 143), 96.54% (977/1 012) and 88.55% (1 021/1 153) in groups A1, A2, B1 and B2 (χ2 = 207.773, P < 0.05), respectively. In addition, there was no significant difference in the overall snail mortality between groups A1 and B1 (P > 0.05), and the snail mortality in groups A1 and B1 were both statistically different from that in groups A2 and B2 (all P values < 0.05). Conclusion Both 50% wettlable powder of niclosamide ethanolamine salt and 5% niclosamide ethanolamine salt granule sprayed with drones are active against O. hupensis snails in ditches, and environmental cleaning may improve the molluscicidal effect.
9.Consistency between bioelectrical impedance analysis and dual-energy X-ray absorptiometry in body composition measurement in children aged 6-17 years in China
Ruihe LUO ; Liping SHEN ; Qian ZHANG ; Wei CAO ; Hongliang WANG ; Peipei XU ; Zhenyu YANG ; Qian GAN ; Xuehong PANG ; Tao XU ; Bowen CHEN ; Yuying WANG ; Wenhua ZHAO
Chinese Journal of Epidemiology 2024;45(11):1507-1512
Objective:To evaluate the consistency between bioelectrical impedance analysis (BIA) and dual-energy X-ray absorptiometry (DXA) in the measurement of body composition in children aged 6-17 years in China.Methods:Fat-free mass, fat mass and fat mass percentage were measured by both BIA and DXA in 1 161 children. t-test or Wilcoxon paired test was used to evaluate the different outcome of the two methods. The correlation and consistency between the methods were evaluated by Spearman correlation coefficients ( r) and Bland-Altman analysis. Results:Body compositions measured by BIA was positively correlated with those measured by DXA (fat mass r=0.95, fat-free mass r=0.98, fat mass percentage r=0.86, all P<0.05). Comparing with DXA, BIA underestimate children's fat mass [the mean difference is -3.15 kg, and the SD is 2.35 kg, 95% limits of agreement (LoA): -7.74-1.45 kg] and fat mass percentage (the mean difference is -8.45%, and the SD is 4.63%, 95% LoA: -17.53%-0.64%). Conclusions:Body compositions measured by BIA was highly positively correlated with those measured by DXA. BIA has certain application value in the measurement of body fat mass and fat-free mass of children aged 6-17 years.
10.Efficacy observation of clinical medication guided by genetic testing of antipsychotic drugs in patients with schizophrenia
Haiyan WANG ; Jianfeng CAO ; Liping FU ; Liangcheng XIANG ; Tao TIAN ; Jixiang WANG ; Ming SHI ; Xiaojun LI ; Chunguang TANG
Sichuan Mental Health 2024;37(5):420-426
Background Results of genetic testing for antipsychotic drugs can guide the rational use of drugs in clinical practice and help improve the clinical symptoms of patients with schizophrenia.However,there is currently limited evidence in China regarding the impact of genetic testing results on medication adherence,social function and drug side effects of antipsychotic drug treatment.Objective To explore the improvement of clinical symptoms,medication adherence and social function in patients with schizophrenia under the guidance of antipsychotic drug gene testing results and examine the safety of drug treatment,so as to provide references for ifor precise treatment of schizophrenia patients.Methods Patients with acute schizophrenia who received hospitalization at Dazhou Minkang Hospital from July 2019 to August 2021 as well as met the diagnostic criteria of the International Classification of Diseases,tenth edition(ICD-10)were selected as research subjects(n=144).Based on random number table,subjects were divided into study group and control group,with 72 cases in each group.Control group received drug treatment based on the doctor's clinical experience,while study group received drug treatment based on the results of gene testing for antipsychotic drug.Both treatments lasted for 12 weeks.At baseline as well as 2,4,8 and 12 weeks after treatment,Positive and Negative Syndrome Scale(PANSS),8-item Morisky Medication Adherence Scale(MMAS-8),Social Functional Rating Scale(SFRS)and Treatment Emergent Symptom Scale(TESS)were adopted for assessment.Result Time effect and group effect of the reducing rate of PANSS,MMAS-8 and SFRS scores in the groups were statistically significant(Ftime=95.251,6.650,14.101,Fgroup=38.055,58.175,128.221,P<0.01).The interaction effect of the reduction rates of MMAS-8 scores in two groups was statistically significant(Finteraction=5.837,P<0.01).The group effect and interaction effect of the severity scores of drug side effects and patient pain scores in two groups were statistically significant(Fgroup=7.553,81.533,Finteraction=8.693,9.322,P<0.01).Conclusion In terms of improving clinical symptom relief,medication adherence,social function and drug side effects,medication for patients with schizophrenia guided by genetic testing of antipsychotic drugs may be more effective than that relying on medication based on clinical experience.

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