1.The mechanism of Prim-O-glucosylcimifugin in improving cholesterol metabolism in osteoarthritis chondrocytes via lncRNA NEAT1/miR-128-3p
Yanming LIN ; Haishui TU ; Shujie LAN ; Chao LI ; Shiyu LU ; Yue CHEN ; Changlong FU
Journal of Beijing University of Traditional Chinese Medicine 2025;48(1):55-67
Objective:
To investigate the mechanism of action of Prim-O-glucosylcimifugin (POG) to improve cholesterol metabolism in osteoarthritic (OA) chondrocytes based on the long noncoding RNA nuclear-enriched transcript 1 (lncRNA NEAT1)/microRNA-128-3p (miR-128-3p) pathway.
Methods:
For in vivo experiments, 60 mice were divided into the normal, sham operation, model, and POG groups using the random number table method, with 15 mice per group. The osteoarthritis mouse model was constructed using the modified Hulth method in the model and POG groups. Mice in the POG group were administered 30 mg/(kg·d)POG by gavage. The other groups were administered an equal amount of normal saline for 8 weeks. The cartilage tissue structure of mice in each group was observed using hematoxylin and eosin staining. Real-time PCR was used to detect changes in the lncRNA NEAT1 and miR-128-3p mRNA expression levels in the cartilage tissues of mice. Western blotting was used to detect the protein expressions of ATP-binding cassette transporter A1 (ABCA1), liver X receptor β (LXRβ), matrix metalloprotein-3 (MMP-3), and B-lymphoblastoma-2-associated X protein (Bax) in articular cartilage of mice. An enzyme-linked immunosorbent assay was used to measure the tumor necrosis factor-α (TNF-α) content in the synovial fluid of mice. A biochemical microplate assay was used to measure the total cholesterol level in the synovial fluid of mice. The in vitro experiments were divided into the negative control, interleukin-1β(IL-1β), IL-1β+ POG, IL-1β+ oe-lncRNA NEAT1, IL-1β+ oe-lncRNA NEAT1 + POG, IL-1β + miR-128-3p inhibition, and IL-1β+ miR-128-3p inhibition+ POG groups. An OA model was established by inducing chondrocytes with IL-1β for 24 h, and 90 mg/L of POG and miR-128-3p inhibitor(50 nmol/L) were administered for 48 h as an intervention. lncRNA NEAT1 expression in chondrocytes was detected using fluorescence in situ hybridization. A dual luciferase assay was used to detect the targeting relationship between lncRNA NEAT1 and miR-128-3p. Lentiviral plasmids overexpressing lncRNA NEAT1 were used to transfect mouse chondrocytes. Real-time PCR was used to detect the effect of lncRNA NEAT1 overexpression on the mRNA level of miR-128-3p in chondrocytes. Western blotting was used to detect ABCA1, LXRβ, MMP-3, and Bax protein expression in chondrocytes after lncRNA NEAT1 overexpression and miR-128-3p inhibition.
Results:
POG significantly reduced OA cartilage tissue damage. Compared with the model group, the lncRNA NEAT1 mRNA level decreased, whereas the miR-128-3p mRNA level increased in the cartilage tissue of the POG group (P<0.05). Compared with the model group, ABCA1 and LXRβ protein expression increased in the POG group, whereas MMP-3 and Bax protein expression decreased (P<0.05). The TNF-α levels decreased in the POG group compared to the model group (P<0.05). Compared with the model group, the total cholesterol level in the synovial fluid of the joint of mice in the POG group decreased (P<0.05). The mean fluorescence intensity of lncRNA NEAT1 in the IL-1β+ POG group decreased compared with the IL-1β group (P<0.05). The relative luciferase activity in the miR-128-3p mimics group bound to the lncRNA NEAT1-WT plasmid decreased compared with the miR-128-3p negative control group (P<0.05). The lncRNA NEAT1 mRNA levels decreased, whereas the miR-128-3p mRNA levels increased in the IL-1β+ oe-lncRNA NEAT1 + POG group compared with the IL-1β+ oe-lncRNA NEAT1 group (P<0.05). Compared with the IL-1β+ POG group, ABCA1 and LXRβ protein expression decreased, whereas MMP-3 and Bax protein expression increased (P<0.05).
Conclusion
POG mediates lncRNA NEAT1/miR-128-3p to improve cholesterol metabolism in OA chondrocytes.
2.Pharmacovigilance for Radiopharmaceuticals
Yue SUN ; Yuxuan ZHENG ; Zhenjiang ZHANG ; Yuxian ZHANG ; Ran ZHANG ; Chang LU ; Li ZHANG ; Ding LI ; Jiachen TU ; Jing XIE ; Huan ZHOU ; Jian GONG
Herald of Medicine 2024;43(10):1615-1619
Radiopharmaceuticals play an important role in the medical field,but they also carry certion risks and potential safety concerns.Medical institutions implement pharmacovigilance to ensure the safety of patients'drug use,including the safety of Radiopharmaceuticals.The operation and management of the pharmacovigilance system in the United States and the European Union are relatively mature.China can learn from their advanced concepts and establish our own radiopharmaciligence system.
3.Design and application of an extracorporeal membrane oxygenation dressing
Lu ZHOU ; Yan YUE ; Shuhan TU ; Min DENG ; Qiulian SONG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2024;31(3):361-363
Extracorporeal membrane oxygenation(ECMO),an effective extracorporeal circulation support system,holds significant importance in the treatment of severe cardiovascular and pulmonary diseases.Effective fixation of ECMO catheters can prevent adverse events such as displacement and accidental extubation,thereby ensuring the effectiveness and continuity of ECMO treatment.At present,various tools such as transparent dressings,surgical dressings,and bandages are commonly used in clinical practice.However,there are problems such as insufficient fixation,unattractive and tidy appearance,and inconvenient replacement.There is still a lack of specific and effective dressings to fix ECMO catheters.Therefore,the medical staff from department of critical care medical of Hospital of Chengdu University of Traditional Chinese Medicine designed ECMO dressing,and obtained the National Utility Model Patent of China(patent number:ZL 202120543021.3),which includes 3 parts:upper dressing,middle dressing,and lower dressing with weak connections in sequence.The top surface of the upper dressing is the upper dressing layer,which is marked with a first scale.The middle part of the bottom surface of the upper dressing layer is fixed with a gauze layer.The top surface of the middle dressing is the middle dressing layer,and the bottom surface of the middle dressing layer is fixedly equipped with a mesh layer along the edges of the 2 length directions.There is a cutting gauze between the 2 mesh layers,and the 2 length edges of the cutting gauze are fixedly connected to the mesh layer.There is a plastic film between the cutting gauze and the middle dressing layer,and cracks are evenly spaced along the length direction of the cutting gauze.The top surface of the middle dressing layer is marked with a second scale.There is an opening at the connection between the lower dressing and the middle dressing,and the bottom surface of the lower dressing is an adhesive layer.This utility model,simple design,good fixing effect,convenient replacement,can avoid catheter slippage and displacement,reduce the occurrence of pressure-related injuries to instruments,and improve the quality of clinical work.It is worth promoting and applying in clinical practice.
4.Identification of Phenolic Acid Derivatives in Danshen Using MS3 and MS2 Spectra Matching Strategy
Han LI ; Ke ZHANG ; Ting LI ; Wei CHEN ; Jun LI ; Peng-Fei TU ; Yun-Fang ZHAO ; Yue-Lin SONG
Chinese Journal of Analytical Chemistry 2024;52(2):267-276,中插19-中插27
"MS/MS spectrum to structure"plays a critical role in the confirmative identification of complicated matrices and is currently regarded as an extremely challenging endeavor.MS/MS information provides vital clues to structural identification.In this study,a strategy was proposed to facilitate unambiguous identification through matching MS3 with MS2 spectra.Initially,MS3 spectra of the featured ions(c-and y-type ions)generated by the decomposition of ester functional group in esters and the MS2 spectrum of the structural unit([M-H]-)were all captured on the Qtrap-MS platform equipped with two tandem-in-space collision cells,including the second quadrupole cell(q2)and linear ion trap(LIT)chambers(actually the third quadrupole unit).Subsequently,the MS/MS spectrum matching between MS3 spectra of the ester compound and MS2 spectra of the structural unit(s)were achieved.As a result,the findings corresponding to MS3 and MS2 spectra matching were summarized.Finally,based on HR-MS/MS information of total salvianolic acid derivatives(TSA),36 kinds of compounds were preliminarily identified through matching with literature information and database retrieval.The applicability of MS3 and MS2 spectra matching strategy was further justified by the confirmative identification of phenolic acid compounds(Rosmarinic acid and salvianolic acid B)in TSA.Above all,MS3 and MS2 spectra matching strategy was quite meaningful towards advancing"MS/MS spectrum to structure"analysis through recognizing and identifying featured fragment ions,and also provided inspiration and new insights for the structural characterization.
5.Analysis of component composition and content determination of six constituents for Xeriga-4 Powder
Jun LI ; Yue-Wu WANG ; Qian ZHANG ; Ping CUI ; Ri-Gui YE ; Ji-ri-mu-ba-tu
Chinese Traditional Patent Medicine 2024;46(1):14-22
AIM To analyze the component composition of Xeriga-4 Powder,and to determine the contents of phellodendrine,chlorogenic acid,gardenoside,berberine,rutin and curcumin.METHODS The high performance liquid chromatography-Q-exactive orbitrap mass spectrometry(HPLC-Q-Exactive-MS)qualitative analysis was performed on a 35℃thermostatic Agilent ZORBAX SB-Aq column(4.6 mm×150 mm,5 μm),with the mobile phase comprising of methanol-0.1%formic acid flowing at 0.35 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning.High performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)quantitative analysis was performed on a 35℃thermostatic Shim-pack GIST-HP C18 column(2.1 mm×100 mm,3 μm),with the mobile phase comprising of methanol-0.1%formic acid flowing at 0.25 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning with multiple reaction monitoring mode.RESULTS Total 65 constituents were identified,containing 19 alkaloids,13 organic acids,13 flavonoids,7 curcumins,6 iridoids,4 fatty acids,2 aldehydes,and 1 amino acid.Six constituents showed good linear relationships within their own ranges(r≥0.999 1),whose average recoveries were 96.44%-102.37%with the RSDs of 2.05%-3.74%.CONCLUSION This study can provide a reference for the quality control for Xieriga-4 Powder.
6.The roles of LMO4 in endothelial cells differentiation and angiogenesis from murine embryonic stem cells
Minghua Xiang ; Zhenzhen Tu ; Yue Wang ; Haisheng Zhou
Acta Universitatis Medicinalis Anhui 2024;59(1):1-7
Objective :
To examine the role of LMO4 in the regulation of endothelial cell differentiation and angio- genesis in murine embryonic stem cells (mESC) .
Methods :
Mouse Lmo4 cDNA was obtained from MEL cells by using the reverse transcription-polymerase chain reaction (RT-PCR) and subcloned into the expression vector pFG to generate the pFLG ,in which contained Flk-1 promoter to drive Lmo4 expresses in only FLK-1 + cells.The mESC were transfected with pFG or pFLG plasmids and subsequently screened with geneticin ( G418) to produce cell clones. These cell clones were named mESC /pFG and mESC /pFLG ,respectively. The mESC /pFG and mESC /pFLG were cultured in the differentiation medium for either 4 days or 10 days to generate embryoid bodies (EB) .The 10-day embryoid bodies ( 10 d-EBs) carrying the pFG and pFLG vectors were subsequently stimulated to generate the blast-colony forming cells (BL-CFC) ,which indicated the presence of hemangioblasts.The endo- thelial cell sprouting analysis was performed by using 10 d-EBs.The expression of the interest genes was detected by using qualitative RT-PCR or Western blot analysis.
Results :
The pFLG expression vector was successfully con- structed through PCR identification.The mESC /pFG and mESC /pFLG cells were obtained after transfected with the pFG or pFLG vectors and selected by G418.The cells spontaneously differentiate to generate EBs,in which some green fluoresce cells were present.Western blot analysis showed that a significant increase in LMO4 expression in both 4 d-EB and 10 d-EB when compared to mESC.BL-CFC analysis showed that the 4 d-EB/ pFLG had a higher cloning efficiency ( 7. 70% ± 1. 27% ) ,comparing with that of the 4 d-EB/ pFG ( 1. 15% ± 0. 48% ) ( P = 0. 021) .Quantitative RT-PCR results showed that the expression of Flk-1,C-kit,Tie-2 and Ve-cad genes in 10 d- EBs /pFLG increased more than 2-fold compared to 10 d-EBs /pFG.The endothelial cell sprouting analysis result showed a significant increase in the number and length of new blood vessels in 10 d-EB/ pFLG compared to 10 d- EB/ pFG (P<0. 05) .
Conclusion
Overexpression of LMO4 promotes hemangioblast differentiation from mESC, and benefits for endothelial cell differentiation and angiogenesis.
7.Risk factors of visceral leishmaniasis in the world: a review
Xi CHEN ; Yue SHI ; Sheng ZHOU ; Mengjie GENG ; Hong TU ; Jiandong SONG ; Canjun ZHENG ; Junling SUN
Chinese Journal of Schistosomiasis Control 2024;36(4):412-421
Visceral leishmaniasis is a zoonotic parasitic disease caused by viscerotropic Leishmania species and transmitted by bites of infected phlebotomine sandflies, which is predominantly prevalent in the Indian subcontinent, eastern Africa and South America. Currently, visceral leishmaniasis is the second most fatal parasitic disease in the world. Because of climate changes, urban development and individual conditions, there are changes in the density of visceral leishmaniasis vector sandflies and the likelihood of contact with humans, resulting in a visceral leishmaniasis transmission risk. The review summarizes natural, social and biological factors affecting the transmission of visceral leishmaniasis, so as to provide insights into formulation of targeted control measures for visceral leishmaniasis.
8.Chiral LC-MS-guided isolation of angular-type pyranocoumarins from Peucedani Radix
Yang YANG ; Xing-cheng GONG ; Peng-fei TU ; Wen-jing LIU ; Yue-lin SONG
Acta Pharmaceutica Sinica 2024;59(8):2343-2349
This study utilized a chiral liquid chromatography-mass spectrometry (LC
9.Effective substances and mechanism of Yishen Guluo Mixture in treatment of chronic glomerulonephritis based on metabolomics and serum pharmacochemistry.
Zhen-Hua BIAN ; Wen-Ming ZHANG ; Jing-Yue TANG ; Qian-Qian FEI ; Min-Min HU ; Xiao-Wei CHEN ; Xiao-Hang YUAN ; Tu-Lin LU
China Journal of Chinese Materia Medica 2023;48(2):492-506
This study aimed to investigate the effective substances and mechanism of Yishen Guluo Mixture in the treatment of chronic glomerulonephritis(CGN) based on metabolomics and serum pharmacochemistry. The rat model of CGN was induced by cationic bovine serum albumin(C-BSA). After intragastric administration of Yishen Guluo Mixture, the biochemical indexes related to renal function(24-hour urinary protein, serum urea nitrogen, and creatinine) were determined, and the efficacy evaluations such as histopathological observation were carried out. The serum biomarkers of Yishen Guluo Mixture in the treatment of CGN were screened out by ultra-performance liquid chromatography-quadrupole time-of-flight/mass spectrometry(UPLC-Q-TOF-MS) combined with multivariate statistical analysis, and the metabolic pathways were analyzed. According to the mass spectrum ion fragment information and metabolic pathway, the components absorbed into the blood(prototypes and metabolites) from Yishen Guluo Mixture were identified and analyzed by using PeakView 1.2 and MetabolitePilot 2.0.4. By integrating metabolomics and serum pharmacochemistry data, a mathematical model of correlation analysis between serum biomarkers and components absorbed into blood was constructed to screen out the potential effective substances of Yishen Guluo Mixture in the treatment of CGN. Yishen Guluo mixture significantly decreased the levels of 24-hour urinary protein, serum urea nitrogen, and creatinine in rats with CGN, and improved the pathological damage of the kidney tissue. Twenty serum biomarkers of Yishen Guluo Mixture in the treatment of CGN, such as arachidonic acid and lysophosphatidylcholine, were screened out, involving arachidonic acid metabolism, glycerol phosphatide metabolism, and other pathways. Based on the serum pharmacochemistry, 8 prototype components and 20 metabolites in the serum-containing Yishen Guluo Mixture were identified. According to the metabolomics and correlation analysis of serum pharmacochemistry, 12 compounds such as genistein absorbed into the blood from Yishen Guluo Mixture were selected as the potential effective substances for the treatment of CGN. Based on metabolomics and serum pharmacochemistry, the effective substances and mechanism of Yishen Guluo Mixture in the treatment of CGN are analyzed and explained in this study, which provides a new idea for the development of innovative traditional Chinese medicine for the treatment of CGN.
Animals
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Rats
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Arachidonic Acid
;
Biomarkers/blood*
;
Blood Proteins
;
Chromatography, High Pressure Liquid
;
Creatinine
;
Drugs, Chinese Herbal/therapeutic use*
;
Glomerulonephritis/metabolism*
;
Metabolomics
;
Urea
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Chronic Disease
;
Disease Models, Animal
;
Complex Mixtures/therapeutic use*
10.Visual analysis of studies on traditional Chinese medicine for prevention and treatment of high-altitude disease
Zhenhui WU ; Huanhua XU ; Yihao WANG ; Bodan TU ; Xianglin TANG ; Maoxing LI ; Yue GAO
Journal of Environmental and Occupational Medicine 2023;40(11):1297-1306
Background Plateau environment may pose a serious impact on the physiological and psychological stress of people stationed on a plateau, especially for those engaged in military training and occupational activities. There is an urgent need to find drugs to prevent and treat injuries caused by high-altitude environment. Objective To analyze the current status, hotspots, and future trends of domestic and international research in the field of prevention and treatment of high-altitude disease (HAD) by traditional Chinese medicine (TCM), and provide references for scientific research. Methods Web of Science Core Collection (WOSCC) and China National Knowledge Infrastructure (CNKI) were searched for literature on TCM and HAD published from inception to 2022. Excel, CiteSpace, VOSviewer, and RStudio softwares were used to conduct visual analysis on the number of publications, types of publications, journals, authors, research institutions, and keywords. Results A total of 501 publications were evaluated in the present study, including 443 Chinese publications and 58 English publications. The annual number of publications showed a rising trend. MA Huiping was the leading author in number of publications in Chinese (37 publications), and ZHANG Yi and MENG Xianli were the leading authors in the number of publications in English (both 8 publications), respectively. The institutions with the most publications in Chinese were The 940th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army and Lanzhou General Hospital of Lanzhou Military Region (both 32 publications), and the institution with the most publications in English was Chengdu University of Traditional Chinese Medicine (8 publications), respectively. The Chinese and English journals with the largest number of publications were the Journal of High Altitude Medicine (39 publications) and the Journal of Ethnopharmacology (10 publications), respectively. The most highly cited Chinese and English literature included Effects of rhodiola on the free radical metabolism and serum creatine kinase after exercise at plateau (61 citations) and Anti-hypoxic activity at simulated high altitude was isolated in petroleum ether extract of Saussurea involucrate (68 citations) , respectively. The most frequent keywords in the Chinese and English literature were high altitude polycythemia and oxidative stress, respectively. The keyword time zone and emergence maps showed that the research hotspots in this field shifted from prevention and treatment of HAD to animal experiments, and then to mechanisms of action, in which oxidative stress, hypoxic injury, inflammation, and apoptosis were the main focuses. Conclusion The research of TCM against HAD is identified from early clinical observation to associations between clinical outcome variation and pharmacological mechanisms, and further to applying multi-omics techniques to explore the physical basis of TCM efficacy and mechanisms of action with focuses like TCM formula and single herb active ingredients, so as to elaborate potential scientific connotation of TCM against HAD.


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