1.Study on the stability of PBMCs recovered from leukocyte-depleted filter residues
Ju LIN ; Zhiqiang XIANG ; Dongfen DU ; Fang YUAN ; Miaoyu WANG ; Yue WU ; Kaiyu HUANG ; Lieyong SANG
Chinese Journal of Blood Transfusion 2025;38(5):723-733
Objective: To identify an optimal back-flush solution for leukocyte-depleted filters that maximizes peripheral blood mononuclear cell (PBMC) recovery with high viability, long-term storage stability, and sterility of the harvested residues, thereby providing a clinically translatable strategy. Methods: Three sterile bag-packaged solutions—Saline, Solvent, and Hanks' balanced salt solution (HBSS)—were used to back-flush randomly assigned leukocyte-depleted filters. Nucleated cell recovery rate and viability of the harvested residues were compared. The optimal solution identified was applied to an expanded sample set. PBMC viability and yield were evaluated after 1h vs 48h storage of the residues. PBMCs isolated from the residues were cryopreserved in liquid nitrogen for 1 month, followed by post-thaw comparisons of viability and T-cell expansion capacity. Results: The Solvent group achieved the highest and most consistent nucleated cell recovery rate. Post-flush recovery rate from filters after 400 mL whole blood processing was (21.3±1.6)% for the Solvent group, significantly higher than Saline group (19.2±6.3)% and HBSS group (11.2±5.0)%, with residues from all groups maintaining viability >90%. No biologically significant difference in residue viability was observed between 48h vs 1h storage groups (93.3±2.3)% vs (95.7±1.8)%). PBMC recovery rates from residues showed no statistical difference between 48h vs 1h storage groups [(48.2%±9.5%)vs (40.41%±8.35%), P>0.05], with (17.7±2.6)×10
cells. After 1-month cryopreservation and 10-day expansion, PBMCs isolated from 48-hour-stored residues retained (91.2±3.2)% viability and achieved a (61.9±15.9)-fold expansion. Conclusion: The bag-packaged Solvent, as a back-flush solution, enables sterile acquisition of leukocyte-depleted filter residues through closed-system tubing connections. These residues maintained PBMC viability and recovery rates after 48h storage at 2℃-8℃, with post-cryopreservation (1-month liquid nitrogen) viability and expansion capacity remaining stable. This protocol complies with blood bank regulatory criteria, addresses the concerns about the infectious window period in cell therapy raw materials, and provides a clinically translatable strategy for PBMC-based applications.
2.Analysis of epidemiological and clinical characteristics of 1247 cases of infectious diseases of the central nervous system
Jia-Hua ZHAO ; Yu-Ying CEN ; Xiao-Jiao XU ; Fei YANG ; Xing-Wen ZHANG ; Zhao DONG ; Ruo-Zhuo LIU ; De-Hui HUANG ; Rong-Tai CUI ; Xiang-Qing WANG ; Cheng-Lin TIAN ; Xu-Sheng HUANG ; Sheng-Yuan YU ; Jia-Tang ZHANG
Medical Journal of Chinese People's Liberation Army 2024;49(1):43-49
Objective To summarize the epidemiological and clinical features of infectious diseases of the central nervous system(CNS)by a single-center analysis.Methods A retrospective analysis was conducted on the data of 1247 cases of CNS infectious diseases diagnosed and treated in the First Medical Center of PLA General Hospital from 2001 to 2020.Results The data for this group of CNS infectious diseases by disease type in descending order of number of cases were viruses 743(59.6%),Mycobacterium tuberculosis 249(20.0%),other bacteria 150(12.0%),fungi 68(5.5%),parasites 18(1.4%),Treponema pallidum 18(1.4%)and rickettsia 1(0.1%).The number of cases increased by 177 cases(33.1%)in the latter 10 years compared to the previous 10 years(P<0.05).No significant difference in seasonal distribution pattern of data between disease types(P>0.05).Male to female ratio is 1.87︰1,mostly under 60 years of age.Viruses are more likely to infect students,most often at university/college level and above,farmers are overrepresented among bacteria and Mycobacterium tuberculosis,and more infections of Treponema pallidum in workers.CNS infectious diseases are characterized by fever,headache and signs of meningeal irritation,with the adductor nerve being the more commonly involved cranial nerve.Matagenomic next-generation sequencing improves clinical diagnostic capabilities.The median hospital days for CNS infectious diseases are 18.00(11.00,27.00)and median hospital costs are ¥29,500(¥16,000,¥59,200).The mortality rate from CNS infectious diseases is 1.6%.Conclusions The incidence of CNS infectious diseases is increasing last ten years,with complex clinical presentation,severe symptoms and poor prognosis.Early and accurate diagnosis and standardized clinical treatment can significantly reduce the morbidity and mortality rate and ease the burden of disease.
3.Effects of total flavonoids of Pterocarya hupehensis Skan on migration,invasion and ferroptosis of non-small-cell lung cancer A549 cells
Guoqing CHEN ; Qiannan DONG ; Rui YANG ; Ying GAO ; Renjia LIU ; Lin YUAN ; Yang XIANG ; Hao WU
Chinese Journal of Pathophysiology 2024;40(2):274-281
AIM:To investigate the impact of total flavonoids of Pterocarya hupehensis Skan(PHSTF)on the migration,invasion,and ferroptosis of non-small-cell lung cancer A549 cells.METHODS:The A549 cells were divided into control group,low-,medium-and high-dose(100,150 and 200 μg/mL)PHSTF groups,ferroptosis inhibitor liprox-statin-1(Lip-1)group,and high-dose PHSTF combined with Lip-1 group,each cultured in corresponding media.Cell via-bility was assessed using the CCK-8 assay,while cell migration and invasion abilities were determined through scratch and Transwell assays.Cell lipid peroxidation levels were measured using the glutathione(GSH)assay kit.RT-qPCR was em-ployed to assess the mRNA expression of solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4),while Western blot was utilized to examine the protein expression of SLC7A11,GPX4,Kelch-like epichlorohy-drin-associated protein-1(Keap-1),nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1).RE-SULTS:Compared with control group,PHSTF significantly diminished the viability of A549 cells in a time-and dose-de-pendent manner(P<0.01),and the cell migration and invasion were also reduced(P<0.01),along with a significant de-crease in GSH level(P<0.01).Treatment with PHSTF inhibited the mRNA and protein expression levels of ferroptosis-re-lated proteins,including SLC7A11 and GPX4(P<0.01),suppressed the protein expression of Nrf2 and HO-1(P<0.01),and enhanced the expression of Keap-1(P<0.01).The Lip-1 partially restored the decrease in cell viability in-duced by PHSTF(P<0.01),significantly up-regulated the protein expression levels of SLC7A11,GPX4,Nrf2 and HO-1,and suppressed the protein expression of Keap-1(P<0.01).CONCLUSION:Total flavonoids of Pterocarya hupehen-sis Skan can inhibit the migration and invasion of non-small-cell lung cancer A549 cells,and induce the cell ferroptosis by regulating the Keap-1/Nrf2/HO-1 pathway.
4.Impact of therapeutic plasma exchange intervention timing and liver injury periodization on the prognosis of pa-tients with exertional heat stroke
Zongzhong HE ; Min WANG ; Yuan ZHUANG ; Jie LIN ; Leiying ZHANG ; Liyang ZOU ; Lingling LI ; Chunya MA ; Xiaomin LIU ; Xiang QUAN ; Ying JIANG ; Mou ZHOU ; Hongjun KANG ; Yang YU
Chinese Journal of Blood Transfusion 2024;37(7):728-733
Objective To explore the prognostic impact and clinical application value of therapeutic plasma exchange(TPE)intervention timing and liver injury periodization in patients with exertional heat stroke(EHS).Methods Data of 127 EHS patients from the First Medical Center of the General Hospital of the People′s Liberation Army from January 2011 to December 2023 were collected,then divided into the death group and the survival group based on therapeutic outcomes and into 5 stages according to the dynamic changes of ALT,AST,TBIL and DBIL.According to propensity score matching analysis,11 patients in the survival group and 12 patients in the death group were included in the statistical analysis,and 20 of them were treated with TPE.The changes in indicators and clinical outcomes before and after TPE were observed,in order to evaluate the impact of intervention timing on prognosis.Results Among the 23 patients,14 had no liver injury or could progress to the repair phase,resulting in 3 deaths(with the mortality rate of 21.43%),while 9 patients failed to pro-gress to the repair phase,resulting in 9 deaths(with the mortality rate of 100%),with significant differences(P<0.05).The mortality rate of the first TPE intervention before the third stage of liver injury was 23.08%(3/13),while that of interven-tion after reaching or exceeding the third stage was 85.71%(6/7),and the difference was statistically significant(P<0.05).Conclusion TPE should be executed actively in EHS patients combined with liver injury before the third phase to lock its pathological and physiological processes,thereby improving prognosis and reducing mortality.
5.Risk factors of allergic reactions caused by therapeutic plasma exchange:a single-center analysis
Lingling LI ; Xiaojun ZHU ; Jie LIN ; Yuan ZHUANG ; Xuede QIU ; Xiang QUAN ; Zongzhong HE ; Ying JIANG ; Yang YU
Chinese Journal of Blood Transfusion 2024;37(7):748-753
Objective To review the occurrence of allergic reactions during therapeutic plasma exchange(TPE)and to explore the risk factors of TPE allergic reactions.Methods The clinical data of 929 patients treated with TPE using plasma components by the Department of Transfusion Medicine in our medical center from 2018 to 2023 were collected.The influen-cing factors of allergic reactions were analyzed by univariate analysis,and the independent risk factors of allergic reactions were analyzed by logistic multivariate regression analysis.Results A total of 4 071 TPEs were performed in 929 patients.A-mong them,198 patients(21.31%)experienced 349 times(8.57%)of allergic reactions,with the incidence of gradeⅠ,Ⅱ and Ⅲ allergic reactions of 16.33%,81.38%and 2.29%,respectively,and no deaths.The univariate analysis showed that the patient′s age,allergy history,diagnosis of immune-related diseases,ICU admission,plasma consumption,total blood volume,maximum blood flow rate and combined use of albumin were related to the occurrence of allergic reactions(P<0.05).Multivariate regression analysis showed that young patients,a history of allergy,immune-related diseases and non-ICU patients were prone to allergic reactions in TPE,but the treatment options of TPE such as substitute fluid category,plasma consumption and blood flow rate were not related to the occurrence of allergic reactions.Conclusion There are sig-nificant individual differences in the occurrence of allergic reactions for TPE,and young age,history of allergies,immune-related diseases and non-ICU patients are risk factors for allergic reactions in TPE.Identifying patients with risk factors be-fore TPE treatment and giving corresponding preventive measures can reduce the incidence of allergic reactions.
6.Comparison of Wild and Cultivated Codonopsis pilosula Based onTraditional Quality Evaluation
Xiaoyan LAN ; Chunfang TIAN ; Zhilai ZHAN ; Li ZHOU ; Xiang LI ; Zidong QIU ; Tiegui NAN ; Qili YUAN ; Xiaoliang LIN ; Congkui TIAN ; Meilan CHEN ; Liping KANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):156-164
ObjectiveTo conduct a systematic comparative study on wild and cultivated Codonopsis pilosula(CP) from three aspects, including characters, microscopy, and contents of primary and secondary metabolites. MethodWild and cultivated CP samples were collected, their characters were measured using vernier caliper, tape measure and balance, the paraffin sections were stained with safranin-fixed green dyeing, and their microstructure were observed under the optical microscope. The content of alcohol-soluble extracts in wild and cultivated CP was determined according to the method for determination of extract under CP in the 2020 edition of Chinese Pharmacopoeia, the starch content was determined by anthrone colorimetry, the content of total polysaccharides was determined by kit method, Fiber analyzer was used to determine the content of fiber components, and ultra performance liquid chromatography(UPLC) was used to determine the content of monosaccharides, disaccharides and some secondary metabolites. Multivariate statistical analysis methods such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were employed to screen key differential components between wild and cultivated CP on the basis of variable importance in the projection(VIP) value>1 and P<0.05. ResultIn terms of morphological characteristics, the "lion's head-like" shape, longitudinal wrinkles, and circumferential wrinkles below the root cap of wild CP were more pronounced in wild CP compared to the cultivated ones. Regarding transverse sectional features, wild CP had more fissures on the outer side of the cortex and a larger duramen. Under microscopic examination, wild CP had more stone cells, a larger proportion of xylem, and the presence of cork cells arranged in rings in the xylem, while cultivated CP has a larger proportion of phloem, smaller vessel diameters, and a more loosely arranged vascular system. In terms of primary metabolites, the contents of 45% ethanol-soluble extract and total polysaccharides in cultivated CP were significantly higher than those in the wild ones(P<0.05), the contents of lignin, hemicellulose, cellulose, fructose and glucose in wild CP were significantly higher than those in the cultivated ones(P<0.05), while sucrose content in the cultivated CP was significantly higher than that in the wild ones(P<0.05). Concerning secondary metabolites, the contents of tryptophan and tangshenoside Ⅰ in cultivated CP were significantly higher than those in the wild ones(P<0.05), whereas the contents of lobetyolinin, lobetyol and atractylenolide Ⅲ in wild CP were significantly higher than those in the cultivated ones(P<0.05). ConclusionThere are significant differences between wild and cultivated CP in terms of morphological characteristics, microscopic features and chemical composition. Glucose, fructose, sucrose, tangshenoside Ⅰ, tryptophan and cellulose components are the key differential components between wild and cultivated CP. Wild CP contains more polyacetylenes and fructose, whereas cultivated CP has higher levels of tangshenoside Ⅰ and sucrose, with noticeably lower cellulose content. These distinctions may be related to their growth conditions, growth years and cultivation techniques. Based on the results of this study, it is recommended to increase polyacetylenes and the content ratio of fructose to sucrose as an indicators to characterize different production methods of CP, in order to guide the high-quality production of CP.
7.Effect of Shexiang Baoxin pill on cardiac angiogenesis in spontaneously hypertensive rats
Rong HUA ; Qing-Hai ZHANG ; Yi TANG ; Qian LI ; Yao-Yuan XIAO ; Lin-Lin LIU ; Ming-Xiang TANG
The Chinese Journal of Clinical Pharmacology 2024;40(8):1180-1183
Objective To explore the effect of Shexiang Baoxin pill on cardiac tissue angiogenesis in spontaneously hypertensive rats(SHR).Method Twenty 12 week old male SHR were randomly divided into experimental group and model group,with 12 week old male SD rats as the normal control group.The experimental group rats were orally administered with Shexiang Baoxin pill(45 mg·kg-1)daily,and their blood pressure was monitored using a non-invasive tail artery blood pressure gauge every four weeks.Eight weeks later,cardiac tissue was taken for platelet endothelial cell adhesion molecule-1(PECAM-1/CD31)immunofluorescence staining to observe CD31 expression level.Use protein blotting to detect the expression levels of myocardial endothelial growth factor(VEGF),myocardial endothelial growth factor receptor 2(VEGF-R2),basic fibroblast growth factor(bFGF)and phosphorylated protein kinase B(p-Akt)/protein kinase B(Akt)proteins.Result There was significant increase in blood pressure between the experimental group,model group and normal group at the same time point(all P<0.01),but there was no statistically significant difference in blood pressure changes between the experimental group and model group at the same time point(all P>0.05).The CD31 expression rates of the normal group,model group and experimental group were(3.79±0.84)%,(2.54±0.42)%and(3.56±0.49)%;VEGF levels were 0.95±0.10,0.73±0.08 and 0.94±0.15;VEGF-R2 levels were 0.85±0.10,0.61±0.14 and 0.80±0.10;bFGF levels were 0.84±0.04,0.51±0.21 and 0.74±0.14;p-Akt/Akt levels were 0.85±0.15,0.57±0.13 and 0.80±0.20,respectively.The differences between the normal group and the model group,as well as the experimental group and the model group,were statistically significant(all P<0.05).Conclusion Shexiang Baoxin pill can promote the neovascularization of microvessels in the heart tissue of spontaneously hypertensive rats,and its mechanism may be related to the activation of PI3K/Akt phosphorylation,upregulation of bFGF,VEGF and their receptor VEGF-R2 in myocardial tissue.
8.Research status of mechanisms of pancreatic islet beta cell injury in type 2 diabetes
Yan-Hui ZHAI ; Yong-Lin LIANG ; Dong AN ; Yuan-Yuan ZHANG ; Fei SU ; Xiao-Wen GUAN ; Xiang-Dong ZHU
The Chinese Journal of Clinical Pharmacology 2024;40(16):2442-2446
The number of type 2 diabetes(T2DM)accounts for more than 90%of all Diabetes mellitus(DM).The decrease of islet β cell mass and dysfunction are the core links of T2DM.With the prolongation of the course of disease,the number and function of β cells are gradually attenuated,and the damage mechanism has not been elucidated.At present,it is believed that it is closely related to metabolic stress,abnormal regulation of insulin secretion,changes in islet microenvironment,mitochondrial damage,glycolipid toxicity and dedifferentiation of islet β cells.Therefore,the mechanism of islet β cell damage in T2DM is summarized and elaborated in order to provide some reference for the precise intervention of T2DM.
9.Biological principles of "food and medicine homologous"
Jin-wen DING ; Xiang-yin CHI ; Yu ZHANG ; Lu-lu WANG ; Jian-dong JIANG ; Yuan LIN
Acta Pharmaceutica Sinica 2024;59(6):1509-1518
With the rapid society development and broad recognition of "Healthy China", the demands for good life and health are increasing. Accordingly, the concept of "food and medicine homologous" have been attractive. The concept of "food and medicine homologous" has a long history in China, and is an essence of various ideas in traditional Chinese medicine, such as diet therapy, medicated diet, regimen and preventive treatment of disease, representing an important field in health science. Many studies have found that the active ingredients of "food and medicine homologous" substances are multiple types, multiple mechanisms and multiple targets, exerting their biological effects after oral administration and chemical or metabolic transformation. In this review, the chemical basis and biological principles of various "food and medicine homologous" substances were summarized as compounds, biological macromolecules and intestinal flora. By focusing on the intestinal flora, we discussed the detailed biological principles of several classic "food and medicine homologous" substances. The scientific significance of "food and medicine homologous" concept were also discussed. This review explores the concept of "food and medicine homologous" from the perspective of modern medicine, in order to provide insights for future drug development and human health.
10.Tujia medicine Toddalia asiatica improves synovial pannus in rats with collagen-induced arthritis through the PI3K/Akt signaling pathway
Shan XIANG ; Zongxing ZHANG ; Lu JIANG ; Daozhong LIU ; Weiyi LI ; Zhuoma BAO ; Rui TIAN ; Dan CHENG ; Lin YUAN
Journal of Southern Medical University 2024;44(8):1582-1588
Objective To investigate the therapeutic mechanism of Tujia medicine Toddalia asiatica alcohol extract(TAAE)for synovial pannus formation in rats with college-induced arthritis(CIA).Methods Sixty male SD rats were randomized into normal control group,CIA model group,TGT group,3 TAAE treatment groups at low,medium and high doses(n=10).Except for those in the normal control group,all the rats were subjected to CIA modeling using a secondary immunization method and treatment with saline,TGT or TAAE by gavage once daily for 35 days.The severity of arthritis was assessed using arthritis index(AI)score,and knee joint synovium pathologies were examined with HE staining.Serum levels of TNF-α,IL-6,and IL-1β were detected with ELISA;the protein expressions of PI3K,Akt,p-PI3K,p-Akt,VEGF,endostatin,HIF-1α,MMP1,MMP3,and MMP9 in knee joint synovial tissues were determined using Western blotting,and the mRNA expressions of TNF-α,IL-6,IL-1β,VEGF,HIF-1α,PI3K,and Akt were detected with RT-PCR.Results Treatment of CIA rat models with TAAE and TGT significantly alleviated paw swelling,lowered AI scores,and reduced knee joint pathology,neoangiogenesis,and serum levels of inflammatory factors.TAAE treatment obviously increased endostatin protein expression,downregulated p-PI3K,p-Akt,MMP1,MMP3,MMP9,VEGF,and HIF-1α proteins,and reduced TNF-α,IL-6,IL-1β,PI3K,Akt,VEGF,and HIF-1α mRNA levels in the synovial tissues,and these changes were comparable between high-dose TAAE group and TGT group.Conclusion TAAE can improve joint symptoms and inhibit synovial pannus formation in CIA rats by regulating the expressions of HIF-1α,VEGF,endostatin,MMP1,MMP3,and MMP9 via the PI3K/Akt signalling pathway.

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