1.Mechanism of Wendantang in Intervention of ApoE-/- Hyperlipidemic Mice Based on Liver Metabolomics
Yun ZHOU ; Songren YU ; Lu ZHANG ; Wenting LIN ; Keming YU ; Min XIA ; Zhijun ZENG ; Yanhua JI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):1-9
ObjectiveTo explore the mechanism of action of Wendantang on ApoE-/- hyperlipidemic mice using non-targeted metabolomics technology. MethodsMale C57BL/6J mice served as the normal control group (n=6), and they were fed with regular chow, while male ApoE-/- mice constituted the high-fat group (n=30), and they were fed with a 60% high-fat diet. After 11 weeks of model establishment, the mice in the high-fat group were randomly divided into the model group, simvastatin group (3.3 mg·kg-1), and high-dose, medium-dose, and low-dose groups of Wendantang (26, 13, 6.5 g·kg-1, respectively, in terms of crude drug amount), with six mice in each group. The normal control group and the model group were gavaged with an equivalent volume of normal saline, and all groups continued to be fed their respective diets, receiving daily medication for 10 weeks with weekly body weight measurements. Serum levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), free fatty acids (NEFA), blood glucose (GLU), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were detected in the mice. Pathological changes in liver tissue were observed using hematoxylin-eosin (HE) staining, and ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) was employed for metabolomic analysis of mouse liver tissue. ResultsCompared to the normal control group, the model group exhibited significantly increased body weight, blood lipid levels, and liver function (P<0.05, P<0.01), with disordered liver tissue structure, swollen hepatocytes, and accompanying vacuolar fatty degeneration and inflammatory cell infiltration. Compared to the model group, the simvastatin group and Wendantang groups showed significantly reduced body weight, TG, NEFA, GLU, ALT, and AST levels (P<0.05, P<0.01), with a significant increase in HDL-C levels (P<0.05, P<0.01), demonstrating a dose-dependent effect. The lesion of the liver tissue section was obviously improved after administration, tending towards a normal liver tissue morphology. Analysis of liver metabolites revealed 86 differential metabolites between the normal control group and the model group, with the high-dose group of Wendantang able to regulate 56 of these metabolites. Twenty-two differential metabolites associated with hyperlipidemia were identified, mainly including chenodeoxycholic acid, hyocholic acid, taurine, glycocholic acid, dihydroceramide, hydroxy sphingomyelin C14∶1, arachidonic acid, and linoleic acid, enriching 22 metabolic pathways, with 4 being the most significant (P<0.05), namely primary bile acid biosynthesis, sphingolipid metabolism, unsaturated fatty acid biosynthesis, and linoleic acid metabolism pathways. ConclusionWendantang can improve blood lipid levels and liver function in ApoE-/- hyperlipidemic mice, which may be related to the regulation of primary bile acid biosynthesis, sphingolipid metabolism, unsaturated fatty acid biosynthesis, and linoleic acid metabolism pathways.
2.Comparative study on the mechanism and efficacy of Zexie tang traditional decoction and formula granules in reducing lipid accumulation
Yuanyuan GUO ; Lina MA ; Huqin LIN ; Changhui ZHENG ; Jiayi LI ; Zhijun LI ; Junling CAO
China Pharmacy 2025;36(10):1202-1208
OBJECTIVE To explore the effect and mechanism of Zexie tang (ZXT) on reducing lipid accumulation through network pharmacology, and compare the difference of traditional decoction versus formula granules. METHODS The active components and targets of ZXT were identified using TCMSP and SwissTargetPrediction databases. GeneCards, OMIM, DisGeNET and TTD databases were used to analyze the related targets of non-alcoholic fatty liver disease (NAFLD); protein-protein interaction network model was constructed by String database;“ ZXT-NAFLD target-pathway” network diagram was constructed by using CytoScape software; target enrichment analysis was performed by using Metascape platform. Fat accumulation model of human hepatocellular carcinoma HepG2 cells was established to observe the effects of traditional decoction and formula granules of ZXT on lipid accumulation of cells. RESULTS Alisol B, alisol C, 1-monolinolein and alisol B monoacetate were the key active components of ZXT in the treatment of NAFLD. The core targets included MDM2, MAPK1, PIK3CB, PRKCQ and MAPK14, etc. The core signaling pathways included endocrine resistance, insulin resistance and Th17 cell differentiation. Compared with model group, except for the Zexie formula granules group and Baizhu formula granules group, the absorbance values in all other administration groups were significantly decreased (P<0.01); the absorbance value of Baizhu traditional decoction group was significantly higher than that of ZXT traditional decoction group (P<0.01); the absorbance values of Zexie formula granule group and Baizhu formula granule group were significantly higher than that of ZXT formula granule group (P<0.01); the absorbance value of Zexie formula granule group was significantly higher than that of Zexie traditional decoction group (P<0.01); the absorbance value of Baizhu formula granule group was significantly higher than that of Baizhu traditional decoction group (P<0.01). CONCLUSIONS ZXT reduces lipid accumulation of human hepatocellular carcinoma cells through multiple components, multiple target and multiple pathways, and its traditional decoction and formula granules exhibit slightly different lipid-lowering effects.
3.Identification of unknown pollutants in drinking water based on solid-phase extraction and supramolecular solvent extraction
Zixin QIAN ; Yuhang CHEN ; Chao FENG ; Yuanjie LIN ; Qian XU ; Ziwei LIANG ; Xinyu WANG ; Dasheng LU ; Ping XIAO ; Zhijun ZHOU
Journal of Environmental and Occupational Medicine 2025;42(7):854-861
Background With the progression of industrialization, an increasing number of emerging contaminants are entering aquatic environments, posing significant threats to the safety of drinking water. Therefore, establishing a system for identifying unknown hazardous factors and implementing safety warning mechanisms for drinking water is of paramount importance. Among these efforts, non-target screening plays a critical role, but its effectiveness is largely constrained by the scope of coverage of sample pre-treatment methods. Objective To integrate modern chromatography/mass spectrometry techniques with advanced data mining methods to develop a non-discriminatory sample pre-treatment method for comprehensive enrichment of unknown contaminants in drinking water, laying a technical foundation for the discovery and identification of unknown organic hazardous factors in drinking water. Methods A non-discriminatory pre-treatment method based on supramolecular and solid-phase extraction was developed. The final target compounds including 333 pesticides, 194 pharmaceuticals and personal care products (PPCPs), and 59 per- and polyfluoroalkyl substances (PFASs) were used for optimizing the pre-treatment method, confirming its coverage. The impacts of different eluents on the absolute recovery rates of target compounds were compared to select the conditions with the highest recovery for sample pre-treatment. The effects of different supramolecular solvents and salt concentrations on target compound recovery were also evaluated to determine the most suitable solvent and salt concentration. Results The solid-phase extraction elution solvents, supramolecular extraction solvents, and salt concentrations were optimized based on the target compound recovery rates. The optimal recovery conditions were achieved using 2 mL methanol, 2 mL methanol (containing 1% formic acid), 2 mL ethyl acetate, 2 mL dichloromethane, hexanediol supramolecular solvent, and 426 mg salt. The detection method developed based on these conditions showed a good linear relationship for all target compounds in the range of 0.1-100.0 ng·mL−1, with R² > 0.99. The method’s limit of detection ranged from 0.01 ng−1 to 0.95 ng−1, and 95% of target compounds were recovered in the range of 20%-120%, with relative standard deviation (RSD) less than 30%, indicating good precision. Conclusion The combined pre-treatment method of solid-phase extraction and supramolecular solvent extraction can effectively enrich contaminants in drinking water across low, medium, and high polarities, enabling broad-spectrum enrichment of diverse trace contaminants in drinking water. It provides technical support for broad-spectrum, high-throughput screening and identification of organic pollutants in drinking water, and also serves as a reference for establishing urban drinking water public safety warning systems.
4.Mutational Signatures Analysis of Micropapillary Components and Exploration of ZNF469 Gene in Early-stage Lung Adenocarcinoma with Ground-glass Opacities.
Youtao XU ; Qinhong SUN ; Siwei WANG ; Hongyu ZHU ; Guozhang DONG ; Fanchen MENG ; Zhijun XIA ; Jing YOU ; Xiangru KONG ; Jintao WU ; Peng CHEN ; Fangwei YUAN ; Xinyu YU ; Jinfu JI ; Zhitong LI ; Pengcheng ZHU ; Yuxiang SUN ; Tongyan LIU ; Rong YIN ; Lin XU
Chinese Journal of Lung Cancer 2024;26(12):889-900
BACKGROUND:
In China, lung cancer remains the cancer with the highest incidence and mortality rate. Among early-stage lung adenocarcinomas (LUAD), the micropapillary (MPP) component is prevalent and typically exhibits high aggressiveness, significantly correlating with early metastasis, lymphatic infiltration, and reduced five-year survival rates. Therefore, the study is to explore the similarities and differences between MPP and non-micropapillary (non-MPP) components in malignant pulmonary nodules characterized by GGOs in early-stage LUAD, identify unique mutational features of the MPP component and analyze the relationship between the ZNF469 gene, a member of the zinc-finger protein family, and the prognosis of early-stage LUAD, as well as its correlation with immune infiltration.
METHODS:
A total of 31 malignant pulmonary nodules of LUAD were collected and dissected into paired MPP and non-MPP components using microdissection. Whole-exome sequencing (WES) was performed on the components of early-stage malignant pulmonary nodules. Mutational signatures analysis was conducted using R packages such as maftools, Nonnegative Matrix Factorization (NMF), and Sigminer to unveil the genomic mutational characteristics unique to MPP components in invasive LUAD compared to other tumor tissues. Furthermore, we explored the expression of the ZNF469 gene in LUAD using The Cancer Genome Atlas (TCGA) database to investigate its potential association with the prognosis. We also investigated gene interaction networks and signaling pathways related to ZNF469 in LUAD using the GeneMANIA database and conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Lastly, we analyzed the correlation between ZNF469 gene expression and levels of immune cell infiltration in LUAD using the TIMER and TISIDB databases.
RESULTS:
MPP components exhibited a higher number of genomic variations, particularly the 13th COSMIC (Catalogue of Somatic Mutations in Cancer) mutational signature characterized by the activity of the cytidine deaminase APOBEC family, which was unique to MPP components compared to non-MPP components in tumor tissues. This suggests the potential involvement of APOBEC in the progression of MPP components in early-stage LUAD. Additionally, MPP samples with high similarity to APOBEC signature displayed a higher tumor mutational burden (TMB), indicating that these patients may be more likely to benefit from immunotherapy. The expression of ZNF469 was significantly upregulated in LUAD compared to normal tissue, and was associated with poor prognosis in LUAD patients (P<0.05). Gene interaction network analysis and GO/KEGG enrichment analysis revealed that COL6A1, COL1A1, COL1A2, TGFB2, MMP2, COL8A2 and C2CD4C interacted with ZNF469 and were mainly involved in encoding collagen proteins and participating in the constitution of extracellular matrix. ZNF469 expression was positively correlated with immune cell infiltration in LUAD (P<0.05).
CONCLUSIONS
The study has unveiled distinctive mutational signatures in the MPP components of early-stage invasive LUAD in the Asian population. Furthermore, we have identified that the elevated expression of mutated ZNF469 impacts the prognosis and immune infiltration in LUAD, suggesting its potential as a diagnostic and prognostic biomarker in LUAD.
Humans
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Lung Neoplasms/genetics*
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Adenocarcinoma of Lung/genetics*
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China
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Prognosis
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Transcription Factors
5.Establishment of high-throughput liquid chromatography tandem mass spectrometry method for determination of 53 per- and polyfluoroalkyl substances in serum
Zheng WANG ; Boya ZHANG ; Jiming ZHANG ; Chao FENG ; Yuanjie LIN ; Chunhua WU ; Dasheng LU ; Zhijun ZHOU
Journal of Environmental and Occupational Medicine 2024;41(4):375-383
Background Per- and polyfluoroalkyl substances (PFAS) are a class of persistent organic pollutants that possess potential toxicity to the human body. The production and utilization of diverse emerging PFAS have resulted in widespread human exposure. Therefore, it is imperative to establish a quantitative methodology encompassing a wide range of PFAS for a comprehensive assessment of human exposure to these compounds. Objective To establish a high-throughput quantitative method for the simultaneous determination of 53 PFAS in human serum based on ultra-high-performance liquid chromatography-Q Exactive high resolution mass spectrometry (UPLC-Q Exactive HRMS). Methods The extraction recoveries of hydrophilic-lipophilic balance (HLB) column, weak anionexchange (WAX) column, and 96-well WAX μElution plate were compared to select the SPE column with the highest recovery. The retention time and peak shape of the target compounds were compared between ACQUITY UPLC BEH C18 column and Accucore aQ column, and the more cost-effective column was chosen. The effects of adding different levels of ammonium formate (0, 2, 5 and 10 mmol·L−1) in mobile phase on peak shape and target response were compared to determine the optimal buffer salt concentration. The optimal spray voltage was obtained by comparing −2 kV and −4 kV. The proposed method was validated from the aspects of selectivity, standard curve, limits of detection, precision, accuracy, and matrix effect. The method was applied to 142 umbilical serum samples. Results The best recovery rate (64%-118%) was achieved by using 96-well WAX μElution plate. The optimal separation and peak shape were obtained by utilizing Accucore aQ column with H2O-methanol (containing 5 mmol·L−1 ammonium formate) as the mobile phase. Less in-source collision and better target response were observed when the spray voltage was set to −2 kV. All target analytes had a good linearity, with R2 > 0.99. The limits of detection ranged from 0.01 to 0.50 μg·L−1, and the recovery ranged from 69% to 127% with the precision less than 26%. A total of 31 PFAS were detected in the 142 actual samples, among which 14 PFAS had a detection frequency over 50%. Perfluorooctanoic acid showed the highest median concentration of 4.16 μg·L−1, followed by 6:2 chlorinated polyfluorinated ether sulfonate and perfluorooctane sulfonates (3.50 μg·L−1 and 1.59 μg·L−1, respectively). Conclusion In this study, we establish a UPLC-Q Excative HRMS method for simutanious determination of 53 PFAS concentrations in serum. This method has the advantages of wide coverage of PFAS, good selectivity, and easy operation, and is suitable for biological detection with a large sample size.
6.Finite element model establishment and stress analysis of lumbar-sacral intervertebral disc in ankylosing spondylitis
Zhijie KANG ; Zhenhua CAO ; Yangyang XU ; Yunfeng ZHANG ; Feng JIN ; Baoke SU ; Lidong WANG ; Ling TONG ; Qinghua LIU ; Yuan FANG ; Lirong SHA ; Liang LIANG ; Mengmeng LI ; Yifei DU ; Lin LIN ; Haiyan WANG ; Xiaohe LI ; Zhijun LI
Chinese Journal of Tissue Engineering Research 2024;28(6):840-846
BACKGROUND:Ankylosing spondylitis is a chronic inflammatory disease with chronic rheumatic immunity.Soft tissue ossification and fusion and spinal stiffness can cause biomechanical changes. OBJECTIVE:To reconstruct the lumbar-sacral intervertebral disc in ankylosing spondylitis patients with lumbar kyphosis by finite element analysis,and to study the range of motion of each segment of T11-S1 and the biomechanical characteristics of annulus fibrosus and nucleus pulposus. METHODS:The imaging data were obtained from an ankylosing spondylitis patient with lumbar kyphosis.The original CT image data of continuously scanned spine were imported into Mimics 21.0 in DICOM format,and T11-S1 was reconstructed respectively.The established model was imported into 3-Matic software in the format of"Stl"to reconstruct the intervertebral disc,and the fibrous intervertebral disc model was obtained.The improved model was further imported into Hypermesh software,and the vertebra,nucleus pulposus,annulus fibrosus and ligament were mesh-divided.After the material properties were given,the model was imported into ABAQUS software to observe the range of motion of each vertebral body in seven different working conditions of T11-S1,and analyze the biomechanical characteristics of each segment of annulus fibrosus and nucleus pulposus. RESULTS AND CONCLUSION:(1)The range of motion of L1 vertebrae was higher than that of other vertebrae under six different working conditions:extension,forward flexion,rotation(left and right),and lateral flexion(left and right).The maximum range of motion was 2.18° during L1 vertebral flexion,and the minimum range of motion was 0.12° during L5 vertebral extension.(2)The annular fiber flexion at L2-L3 segments was greater than the extension(P<0.05),and the annular fiber flexion at L3-L4 and L4-L5 segments was less than the extension(P<0.05).The left rotation of L1-L2 annular fibers was greater than the right rotation(P<0.05).The left flexion of the annulus was greater than the right flexion in L1-L2,L2-L3,L3-L4,L4-L5 and L5-S1 segments(P<0.05).(3)The nucleus pulposus stresses of T11-L12,L1-L2,L2-L3,L3-L4 and L4-L5 segments in forward flexion were greater than in extension(P<0.05).The left rotation of T12-L1 and L3-L4 segments was smaller than the right rotation(P<0.05),and that of T11-T12,L1-L2,and L2-L3 segments was larger than the right rotation(P<0.05).The left flexion was larger than the right flexion in the T11-S1 segment.(4)It is concluded that in ankylosing spondylitis patients with lumbar kyphosis,the minimum range of motion of the vertebral body is located at the L5 vertebral body in extension.To prevent fractures,it is recommended to avoid exercise in the extension position.During the onset of lumbar kyphosis in patients with ankylosing spondylitis,the maximum stress of the annulus fibrosus and nucleus pulposus is located in the L1-L2 segment,which is fixed and will not alter with the change of body position.The late surgical treatment and correction of deformity should focus on releasing the pressure of the annulus fibrosus and nucleus pulposus in this segment to avoid the rupture of the annulus fibrosus and the injury of the nucleus pulposus.
7.Pharmacological Effect of Nuanxinkang Powder on Ventricular Remodeling in Post-infarction Mice Through"Metabolic-Inflammatory"Network Regulating Macrophage Polarization
Zhijun LIN ; Zixin CHEN ; Jialin JIANG ; Xin DONG ; Zhuoji GUAN ; Lingjun WANG
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(2):159-167
Objective To explore the mechanism of Nuanxinkang Powder(aka.NXK,composed of Ginseng Radix et Rhizoma Rubra and Ilex Pubescens Radix)on improving ventricular remodeling in post-infarction mice based on the"metabolic-inflammatory"network regulating macrophage polarization.Methods ①Thirty C57BL/6J male mice were randomly divided into three groups:sham-operation group,model group,and NXK group(1.65 g·kg-1),with 10 mice in each group;the mouse model of myocardial infarction was replicated using left anterior descending coronary artery ligation;and the drug was administered by gavage once a day for 4 consecutive weeks.Masson staining was used to detect collagen deposition in myocardial tissue;ultrasound was used to detect cardiac function in mice:left ventricular ejection fraction(LVEF),left ventricular anterior wall thickness at end-systole(LVAWS)and left ventricular anterior wall thickness at end-diastole(LVAWD);flow cytometry was used to detect distribution of cardiac macrophages in mice;qPCR was used to detect mRNA expressions of lactate dehydrogenase A(LDHA),carnitine palmitoyltransferase 1(CPT-1),glucose transport protein 4(GLUT4),isocitrate dehydrogenase(IDH),and succinate dehydrogenase(SDHa)in heart tissue.②NXK was given 1.15 g·kg-1 NXK suspension to rats by gavage twice a day for 5 consecutive days to prepare NXK-containing serum.Lipopolysaccharide(LPS)-induced RAW 264.7 cells were used to construct a pro-inflammatory macrophage model.The cells were grouped into the following groups:blank serum control group(medium containing 5%blank serum+5%fetal bovine serum),NXK drug-containing serum group(medium containing 5%NXK drug-containing serum+5%fetal bovine serum),lipopolysaccharide group(medium containing 5%blank serum+5%fetal bovine serum+200 μg·mL-1 lipopolysaccharide),NXK drug-containing serum+ lipopolysaccharide group(medium containing 5%NXK drug-containing serum+5%fetal bovine serum+200 μ g·mL-1 lipopolysaccharide),all the groups were intervened for 16 hours.Glycolysis stress test was used to detect the level of glycolysis in RAW 264.7 cells;qPCR was used to detect the mRNA expression of mitochondrial pyruvate carrier(MPC1)in RAW 264.7 cells;and MitoSox Red fluorescent staining was used to detect the level of oxidative stress damage in mitochondria of RAW 264.7 cells.Results ①Compared with the sham-operation group,the blue-stained area of cardiac collagen fibres in mice of the model group was significantly increased,accompanied by thinning of the ventricular wall and enlargement of the left ventricular cavity;cardiac function indexes,such as LVEF,LVAWS,LVAWD,etc.,were all significantly reduced(P<0.01,P<0.001);the mRNA expressions of LDHA and CPT-1 were significantly up-regulated in the cardiac tissues of mice(P<0.05),and the mRNA expressions of GLUT4,IDH and SDHa were significantly down-regulated(P<0.05,P<0.01),and CD86 staining positive cell was significantly increased(P<0.001).Compared with the model group,mice in the NXK group showed a significant decrease in cardiac collagen fiber deposition and an increase in the thickness of the ventricular wall;cardiac function indexes such as LVEF,LVAWS and LVAWD were significantly increased(P<0.05,P<0.01,P<0.001);and the mRNA expressions of LDHA and CPT-1 in the cardiac tissues of the mice were significantly down-regulated(P<0.01,P<0.001),mRNA expressions of GLUT4,SDHa and IDH were significantly up-regulated(P<0.01),and the number of CD86 positive cells was significantly reduced(P<0.001).②Compared with the blank serum control group,the cytosolic glycolysis level and ROS level of macrophages in the NXK serum-containing group did not change significantly(P>0.05),whereas the glycolysis level and ROS level of macrophages in the lipopolysaccharide group were significantly increased(P<0.01),and the mRNA expression of MPC1 was significantly down-regulated(P<0.001).Compared with the lipopolysaccharide group,the macrophage glycolysis level and ROS level were significantly reduced in the NXK serum-containing + lipopolysaccharide group(P<0.05,P<0.01),and mRNA expression of MPC1 was significantly up-regulated(P<0.001).Conclusion NXK can reduce myocardial fibrosis and ventricular remodeling after myocardial infarction and improve cardiac function in mice,and its mechanism may be related to the down-regulation of mRNA expression of LDHA in cardiac tissues,the up-regulation of mRNA expression of GLUT4,the improvement of cardiac glucose uptake after myocardial infarction,the inhibition of pro-inflammatory macrophage glycolysis,the increase in the expressions of SDHa and IDH to alleviate the accumulation of succinate and citrate,and the reduction of reactive oxygen species(ROS)generation,thereby reducing pro-inflammatory macrophage hyperpolarisation.
8.Mechanism Evolution of Latent Toxin in Systemic Lupus Erythematosus and Syndrome Differentiation and Treatment
Yi ZHANG ; Zhijun XIE ; Lin HUANG ; Qiao WANG ; Haichang LI ; Chengping WEN
Journal of Traditional Chinese Medicine 2024;65(16):1721-1724
It is proposed that the disease mechanism evolution of systemic lupus erythematosus can be summarized into four stages: initial invasion and latency, the pathogenesis remains concealing; latent toxin accumulation, the disease gradually becomes apparent; active toxin begins damaging, the disease manifests aggressively; damage resulting to deficiency, the disease course prolonged. Based on the stages of latent toxin evolution, the syndrome differentiation and treatment of systemic lupus erythematosus can be summarized as follows: during the initial latent stage, characterized by latent dampness and heat stagnation, modified Sanren Decoction (三仁汤) should be used; in the toxin outbreak stage, marked by intense heat toxin, modified Xijiao Dihuang Decoction (犀角地黄汤) combined with modified Qingwen Baidu Decoction (清瘟败毒饮) should be used; during the toxin damage stage, which presents as latent toxin damaging zang-fu organs, modified Qinghao Biejia Decoction (青蒿鳖甲汤) should be used; in the healthy qi deficiency stage, characterized by deficiencies of qi, blood, yin, and yang, modified Xieli Shiquan Ointment (燮理十全膏) should be used.
9.Feasibility of preparing human plasminogen by full chromatography from precipitation of Fraction Ⅲ in low temperature ethanol method
Jin ZHANG ; Shenglan YUE ; Chen ZHU ; Yan PENG ; Yanxiang ZHOU ; Lianzhen LIN ; Kejin CHEN ; Lu FENG ; Yong HU ; Zhijun ZHOU
Chinese Journal of Blood Transfusion 2024;37(11):1293-1300
[Objective] To determine the feasibility of preparing plasminogen (Pg) with Fraction Ⅲ precipitation (hereinafter referred to as FⅢ-P) from low-temperature ethanol process by full chromatography (hereinafter referred to as FⅢ-P process). [Methods] The FⅢ-P was diluted with dissolution buffer at different dilution times and stirring time. The potency and antigen concentration of Pg in dissolution sample were detected and the dissolution and clarification conditions were determined. Pre-treatment of loading sample and pre-experiment of affinity chromatography were carried out on the FⅢ-P dissolution sample to judge whether the loading sample had an impact on the chromatography by observing the performance of the affinity chromatography column and to evaluate whether the affinity chromatography could achieve the purpose of purifying Pg by detecting the plasma protein antigen concentration and Pg potency of the samples in the process. Two batches of FⅢ-P process were studied step by step, and the specific activity, steps and total recovery, and the output of Pg per ton of plasma were calculated. The feasibility of preparing Pg by FⅢ-P process was evaluated by comparing with the data of full chromatography process using plasma as raw material (hereinafter referred to as plasma process). [Results] The FⅢ-P was dissolved with 10 times of dissolution buffer, stirred for 1 hour, centrifuged at room temperature of 10 000×g for 15 minutes. The supernatant was first filtered with a screen, then clarified with an 8/0.8 μm filter, and finally filtered with a 0.45/0.2 μm filter and loaded. Pre-test showed that from clarification and filtration to Pg affinity chromatography, the step recovery of activity and antigen was 39.51% and 108.64%, respectively, the antigen concentration of Pg increased by 31.16 times and the activity increased by 11.39 times after affinity chromatography, which reaching the effect of affinity chromatography purification of Pg. The results of 2 batches of step-by-step scale-up FⅢ-P process showed that the total recoveries of antigen and activity from plasma to SP chromatography of FⅢ-P process were (45.76±1.10)% and (24.15±0.59)%, respectively, which had a total loss of about 1/3 of antigen and about 2/3 of activity compared to the plasma process. The Pg specific activity of SP chromatography eluent was (4.68±0.25) U/mg, which was about half of that of plasma process, but meeted the internal standard of > 4 U/mg. The output of Pg antigen per ton of plasma in the FⅢ-P process was 68.73% of that in the plasma process, and the output of Pg activity per ton of plasma in the plasma process was 29.82% of that in the plasma process, which basically achieved the purpose of waste utilization of FⅢ-P. [Conclusion] The technical route of preparing Pg from FⅢ-P by full chromatography is feasible.
10.Real-time evaluation of pulmonary ventilation of HP movements based on electrical impedance tomography:an observational study
Zhijun GAO ; Hang WANG ; Chunchen WANG ; Yilong QIN ; Xinsheng CAO ; Lin YANG
Journal of Army Medical University 2024;46(16):1882-1889,1896
Objective To explore the feasibility of electrical impedance tomography(EIT)for real-time and accurate monitoring of respiration during HP anti-G movements and the key parameters of pulmonary ventilation.Methods Twelve healthy male students in our university were enrolled in September 2023 and subjected in this study.HP anti-G movements were performed 3 times each for 30 s in the anti-G physiological training apparatus,during which EIT and ophthalmic horizontal arterial pressure were measured to analyze the relationship of the global and local parameters of pulmonary ventilation,including inspiratory volume(IV),expiratory uniform(EU),expiratory speed(ES),center of ventilation(COV)and right-to-left lung ventilation ratio(RtoL)with anti-G ability of anti-G straining maneuver(AGSM).Results The average eye horizontal systolic blood pressure(SBP)at the eye level was 148.82±22.75 mmHg during HP anti-G movements,which was significantly higher than that during quiet breathing(PJ)(95.17±8.51 mmHg,P<0.001).From the global pulmonary ventilation,the participants had significantly increased IV during HP anti-G movements(P<0.001).According the d value(mean increase of eye horizontal SBP=SBPHP-SBPPJ),the subjects were divided into 3 groups,with the d value of>60,30~60 and<30 mmHg,respectively.The inspiratory volume ratio(IVHP/IVPJ)was the highest in the>60 mmHg group and the smallest in the<30 mmHg group(P<0.01).The subjects had significantly decreased EU and more evenly expiration(P<0.05),but no change was seen in the expiratory uniformity ratio(EUHP/EUPJ)among the 3 groups.ES was obviously faster during HP anti-G movements(P<0.001),and the expiratory speed ratio(ESHP/ESPJ)had no significant difference among the 3 groups.The inspiratory time and expiratory time were 0.77±0.32 and 1.59±0.21 s,respectively,and both of them were notably shorter during HP anti-G movements(P<0.001,P<0.01).From the local pulmonary ventilation,COV during HP anti-G movements was significantly smaller than that during PJ(P<0.001),and the ventilation center deviated to the ventral side,and RtoL was decreased and the ventilation distribution deviated to the left lung(P<0.05).Conclusion EIT can perform real-time imaging of pulmonary global and local ventilation during HP anti-G movements,and it has a great application prospect in AGSM training and monitoring.

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