1.Mechanism of chlorogenic acid in mitophagy and inflammation of foam cells based on PINK 1/Parkin pathway
Hong-Hong YU ; Yun-Qi YANG ; Pei LUO ; Qi YU ; Yu-Ling MA ; Wei-Yi TIAN
Chinese Pharmacological Bulletin 2024;40(5):914-920
Aim To investigate the regulatory effects of chlorogenic acid(CGA)on mitochondrial autophagy and inflammation in ox-LDL induced foam cells through PINK1/Parkin signaling pathway.Methods RAW264.7 macrophages were divided into the control group,model group,CGA-L group,CGA-M group,CGA-H group and CGA-H+Mdi group.Oil red O method was used to identify cell foam.The intervention concentration of CGA was determined by CCK-8 meth-od.The mitochondrial structure was observed by trans-mission electron microscope.The expression of IL-1 β,IL-18 and IFN-γ was detected by ELISA.The expres-sion of genes and proteins associated with PINK1/Par-kin mitochondrial autophagy pathway was detected by qRT-PCR and immunofluorescence labeling.Results Oil red O staining showed that the foam cell model was successfully prepared.Compared with the model group,mitochondrial damage was significantly reduced after CGA intervention at different concentrations,and the expressions of PINK1,Parkin,p-Parkin,PHB2 and LC3 Ⅱ were induced,while the expression of TOMM20 was inhibited.The expressions of IL-1 β,IL-18 and IFN-γ decreased(P<0.05 or P<0.01).Compared with the CGA-H group,the CGA-H+Mdi group significantly increased mitochondrial damage,in-hibited the expression of PINK1,Parkin,p-Parkin,PHB2,LC3 Ⅱ,induced the expression of TOMM20,and enhanced the expression of IL-1 β,IL-18,IFN-γ(P<0.01).Conclusions CGA can induce mitochon-drial autophagy in foam cells by regulating PINK1/Par-kin pathway and inhibit the overexpression of pro-in-flammatory factors IL-1 β,IL-18 and IFN-y,which may be one of the anti-atherosclerosis mechanisms of CGA.
2.Detection of the Biological Activity of Interleukin-11 Based on the Reporter Gene Method
Yi-Ying WANG ; Xiao-Ling ZHOU ; Zi-Hong YE ; Ya-Fen ZHANG
Chinese Journal of Biochemistry and Molecular Biology 2024;40(10):1462-1470
Interleukin-11(IL-11)is a multifunctional cytokine that plays a crucial role in various bio-logical processes,including the promotion of hematopoiesis,regulation of the immune system,and facili-tation of tissue repair.These functions highlight its significant importance in both medical research and clinical therapy.Consequently,the precise detection and assessment of IL-11's biological activity are es-sential.The currently employed cell proliferation inhibition assay is cumbersome,time-consuming,and susceptible to interference from Interleukin-6.Here we study the JAK-STAT signaling pathway,a key mechanism of IL-11 actions.We initially establish a stable monoclonal cell line that expresses a luciferase reporter gene responsive to IL-11.We subsequently optimized critical parameters,including the pre-dilu-tion concentration and gradient of IL-11,cell inoculum size,and IL-11 incubation duration.Comprehen-sive validation was undertaken to evaluate accuracy,precision,specificity,stability,and concordance with pharmacopeial methods,ultimately leading to the establishment of a novel reporter gene-based ap-proach for detecting the biological activity of IL-11.This innovative method significantly enhances the de-tection accuracy and sensitivity while reducing the testing time,thereby offering promising prospects for broad applications.
3.Genetic Variation of SH2B3 in Patients with Myeloid Neoplasms
Qiang MA ; Rong-Hua HU ; Hong ZHAO ; Xiao-Xi LAN ; Yi-Xian GUO ; Xiao-Li CHANG ; Wan-Ling SUN ; Li SU ; Wu-Han HUI
Journal of Experimental Hematology 2024;32(4):1186-1190
Objective:To observe the genetic variation of SH2B3 in patients with myeloid neoplasms.Methods:The results of targeted DNA sequencing associated with myeloid neoplasms in the Department of Hematology,Xuanwu Hospital,Capital Medical University from November 2017 to November 2022 were retrospectively analyzed,and the patients with SH2B3 gene mutations were identified.The demographic and clinical data of these patients were collected,and characteristics of SH2B3 gene mutation,co-mutated genes and their correlations with diseases were analyzed.Results:The sequencing results were obtained from 1 005 patients,in which 19 patients were detected with SH2B3 gene mutation,including 18 missense mutations(94.74%),1 nonsense mutation(5.26%),and 10 patients with co-mutated genes(52.63%).Variant allele frequency(VAF)ranged from 0.03 to 0.66.The highest frequency mutation was p.Ile568Thr(5/19,26.32%),with an average VAF of 0.49,involving 1 case of MDS/MPN-RS(with SF3B1 mutation),1 case of MDS-U(with SF3B1 mutation),1 case of aplastic anemia with PNH clone(with PIGA and KMT2A mutations),2 cases of MDS-MLD(1 case with SETBP1 mutation).The other mutations included p.Ala567Thr in 2 cases(10.53%),p.Arg566Trp,p.Glu533Lys,p.Met437Arg,p.Arg425Cys,p.Glu314Lys,p.Arg308*,p.Gln294Glu,p.Arg282Gln,p.Arg175Gln,p.Gly86Cys,p.His55Asn and p.Gln54Pro in 1 case each.Conclusion:A wide distribution of genetic mutation sites and low recurrence of SH2B3 is observed in myeloid neoplasms,among of them,p.Ile568Thr mutation is detected with a higher incidence and often coexists with characteristic mutations of other diseases.
4.Clinical Significance of the Levels of Peripheral Blood Tregs and Cytokines IL-35,TGF-β and IL-10 in Hemophilia A Patients with FⅧ Inhibitor
Hong-Xia HE ; Yan-Yan XIE ; Qing-Yun SUN ; Lin-Hong WANG ; Yi-Wen ZHU ; JIE LI ; Xin WANG ; Zhao-Ling DENG ; Mei-Rong YANG ; Zhen-Yu YAN
Journal of Experimental Hematology 2024;32(4):1197-1200
Objective:To explore the levels of regulatory T cells(Tregs)and cytokines IL-35,TGF-β and IL-10 in peripheral blood of hemophilia A(HA)patients with F Ⅷ inhibitor and their clinical significance.Methods:43 HA patients admitted to the Hematology Department of the Affiliated Hospital of North China University of Science and Technology from October 2019 to December 2020 were selected,including 6 cases with F Ⅷ inhibitor and 37 cases without FⅧ inhibitor.In addition,20 healthy males who underwent physical examinations were selected as healthy controls.Flow cytometry was used to detect the levels of CD4+CD25+CD127-Tregs in peripheral blood of the HA patients and healthy controls,and ELISA assay was used to detect the expression levels of IL-35,TGF-β and IL-10 in serum,and their differences between different groups were compared.Results:Compared with the healthy control group,the level of Tregs in HA patients was decreased,and the level of Tregs in the FⅧ inhibitor positive group was the lowest,the difference was statistically significant(P<0.05).There was no significant difference in the expression level of Tregs in HA patients of different severity levels.The serum IL-35,TGF-β,and IL-10 levels in both FⅧ inhibitor negative and positive groups were significantly lower than those in healthy control group,and those in FⅧ inhibitor positive group were significantly lower than those in FⅧ inhibitor negative group(all P<0.05).Conclusion:The decrease of Tregs,IL-35,TGF-β,and IL-10 levels in HA patients may be related to the formation of FⅧ inhibitors.
5.Disease acceptance in HIV/AIDS patients and related factors
Zi-Qi QIN ; Gui-Ying CAO ; Jian-Ping XIE ; Xiao WANG ; Yi-Xuan LI ; Qiao-Yue LU ; Hong-Hong WANG ; Xue-Ling XIAO
Chinese Journal of Infection Control 2024;23(8):1016-1022
Objective To understand the disease acceptance status and related factors in human immunodeficiency virus(HIV)-infected/acquired immunodeficiency syndrom(AIDS)patients,so as to guide the clinical development of intervention measures,and to provide empirical evidence for improving clinical outcomes.Methods Convenience sampling method was used to select 555 HIV-infected/AIDS patients who received treatment in the designated AIDS treatment clinic of a hospital.General data,disease acceptance,disease self-management efficacy and clinical out-comes(such as quality of life,CD4+T lymphocyte count and HIV viral load)of the studied subjects were collected.Results The average disease acceptance of HIV-infected/AIDS patients was(26.08±5.34)points.Multiple linear regression analysis showed that religious belief and self-management efficacy were related factors affecting the di-sease acceptance of patients(both P<0.05),which could explain the 30.4%variation in disease acceptance of HIV-infected/AIDS patients,and the disease acceptance of patients was closely related to their quality of life(P<0.001).Conclusion HIV-infected/AIDS patients have a moderate level of disease acceptance.Medical staff should fully consider patients'religious beliefs and self-management efficacy,so as to formulate targeted intervention mea-sures to improve patients'acceptance of disease,and further promote patients'quality of life.
6.Recent advances in lamellar liquid crystal emulsification methods encapsulating natural active substances for functional cosmetics
Yi ZHANG ; Wei CHEN ; Yan-qi HAN ; Qian-wen SUN ; Yue GAO ; Jun YE ; Hong-liang WANG ; Li-li GAO ; Yu-ling LIU ; Yan-fang YANG
Acta Pharmaceutica Sinica 2024;59(2):350-358
Due to the high similarity with the lipid layer between human skin keratinocytes, functional cosmetics with layered liquid crystal structure prepared by liquid crystal emulsification technology encapsulating natural active substances have become a hot research topic in recent years. This type of functional cosmetic often has a fresh and natural skin feel, excellent skin barrier repair function and efficient moisturizing effect, etc., showing great potential in cosmetic application. However, the present research on the application of liquid crystal emulsification technology to functional cosmetics is still in the initial stage, and there are fewer relevant reports with reference values. Based on the mentioned above, this review provides a comprehensive summary of functional cosmetics with layered liquid crystal structures prepared by liquid crystal emulsification technology from the following aspects: the structure of human skin, the composition of lamellar liquid crystal, the advantages of liquid crystal emulsification technology containing natural active substances used in the field of functional cosmetics, the preparation process, main components, influencing factors during the preparation and the market functional cosmetics with lamellar liquid crystal structure. Finally, the prospect of the application of liquid crystal emulsification technology in functional cosmetics is presented, to provide useful references for those engaged in the research of liquid crystal emulsification technology-related functional cosmetics.
7.Latent tuberculosis infection among close contacts of positive etiology pul-monary tuberculosis in Chongqing
Rong-Rong LEI ; Hong-Xia LONG ; Cui-Hong LUO ; Ben-Ju YI ; Xiao-Ling ZHU ; Qing-Ya WANG ; Ting ZHANG ; Cheng-Guo WU ; Ji-Yuan ZHONG
Chinese Journal of Infection Control 2024;23(3):265-270
Objective To investigate the current situation and influencing factors of latent tuberculosis infection(LTBI)among close contacts of positive etiology pulmonary tuberculosis(PTB)patients,provide basis for formula-ting intervention measures for LTBI.Methods A multi-stage stratified cluster random sampling method was used to select close contacts of positive etiology PTB patients from 39 districts and counties in Chongqing City as the study objects.Demographic information was collected by questionnaire survey and the infection of Mycobacterium tuberculosis was detected by interferon gamma release assay(IGRA).The influencing factors of LTBI were analyzed by x2 test and binary logistic regression model.Results A total of 2 591 close contacts were included,the male to female ratio was 0.69∶1,with the mean age of(35.72±16.64)years.1 058 cases of LTBI were detected,Myco-bacterium tuberculosis latent infection rate was 40.83%.Univariate analysis showed that the infection rate was dif-ferent among peoples of different age,body mass index(BMI),occupation,education level,marital status,wheth-er they had chronic disease or major surgery history,whether they lived together with the indicator case,and whether the cumulative contact time with the indicator case ≥250 hours,difference were all statistically significant(all P<0.05);infection rate presented increased trend with the increase of age and BMI(both P<0.001),and decreased trend with the increase of education(P<0.05).Logistic regression analysis showed that age 45-54 years old(OR=1.951,95%CI:1.031-3.693),age 55-64 years old(OR=2.473,95%CI:1.279-4.781),other occupations(OR=0.530,95%CI:0.292-0.964),teachers(OR=0.439,95%CI:0.242-0.794),students(OR=0.445,95%CI:0.233-0.851),junior high school education or below(OR=1.412,95%CI:1.025-1.944),BMI<18.5 kg/m2(OR=0.762,95%CI:0.586-0.991),co-living with indicator cases(OR=1.621,95%CI1.316-1.997)and cumu-lative contact time with indicator cases ≥250 hours(OR=1.292,95%CI:1.083-1.540)were the influential fac-tors for LTBI(all P<0.05).Conclusion The close contacts with positive etiology PTB have a high latent infection rate of Mycobacterium tuberculosis,and it is necessary to pay attention to close contacts of high age,farmers,and frequent contact with patients,and take timely targeted interventions to reduce the risk of occurrence of disease.
8.Simultaneous content determination of seventeen constituents in Yangxue Ruanjian Capsules by UPLC-MS/MS
Yong-Ming LIU ; Shu-Sen LIU ; Yi-Zhe XIONG ; Xiang WANG ; Yu-Yun WU ; Jin LIU ; Ling-Yun PAN ; Guo-Qing DU ; Hong-Sheng ZHAN
Chinese Traditional Patent Medicine 2024;46(2):353-358
AIM To establish a UPLC-MS/MS method for the simultaneous content determination of liquiritin apioside,alibiflorin,swertiamarin,methyl gallate,benzoylpaeoniflorin,sweroside,6′-O-β-D-glucosylgentiopicroside,isoliquiritigenin,loganic acid,liquiritigenin,gallic acid,paeoniflorin,oxypaeoniflorin,gentiopicroside,glycyrrhizic acid,isoliquiritoside and liquiritin in Yangxue Ruanjian Capsules.METHODS The analysis was performed on a 40℃thermostatic Waters BEH C18column(2.1 mm×100 mm,1.7 μm),with the mobile phase comprising of 2 mmol/L ammonium acetate(containing 0.1%formic acid)-acetonitrile flowing at 0.3 mL/min in a gradient elution manner,and electron spray ionization source was adopted in negative ion scanning with multiple reaction monitoring mode.RESULTS Seventeen constituents showed good linear relationships within their own ranges(r>0.999 6),whose average recoveries were 91.33%-104.03%with the RSDs of 1.58%-3.50%.CONCLUSION This rapid,accurate and stable method can be used for the quality control of Yangxue Ruanjian Capsules.
9.Effects of rhynchophylline on methamphetamine-dependent SH-SY5Y cells model and expression of miR-375-3p/Elavl4
Han-Cheng LI ; Ming ZENG ; Yi-Ling CHEN ; Ling-Hong CHEN ; Zhi-Xian MO
The Chinese Journal of Clinical Pharmacology 2024;40(1):47-51
Objective To investigate the effects of rhynchophylline on methamphetamine-dependent SH-SY5Y cells model and microRNA-375-3p(miR-375-3p)/embryonic lethal abnormal vision drosophila-like 4(Elavl4)expression.Methods A methamphetamine-dependent cell model by maximum safe dose induction was established.The cells were divided into normal group(complete culture medium),control group(complete culture medium+400 μmol·L-1 rhynchophylline incubated for 48 h),model group(complete culture medium+100 μmnol·L-1 methamphetamine incubated for 48 h)and experimental group(complete culture medium+400 μmol·L-1 rhynchophylline incubated for 15 min,then 100 μmol·L-1 methamphetamine incubated for 48 h).The cyclic adenosine monophosphate(cAMP)and 5-hydroxytryptamine(5-HT)expression were detected by enzyme-linked immunosorbent assay(ELISA),miR-375-3p expression was detected by quantitative real time polymerase chain reaction(qRT-PCR),and Elavl4 expression was detected by Western blot.Results The cAMP expression levels of normal group,control group,model group and experimental group were(6.33±0.93),(6.57±1.12),(10.89±1.03)and(7.81±1.32)pmol·mg-1;5-HT were(682.46±17.32),(690.31±15.09),(510.11±27.67)and(649.99±21.42)pg·mL-1;miR-375-3p expression were 1.00±0.13,1.13±0.24,3.48±0.18 and 1.58±0.19;Elavl4 expression were 1.00±0.05,0.89±0.10,0.50±0.09 and 0.90±0.11,respectively.The differences between above indicators in model group and normal group were statistically significant(P<0.01,P<0.05);the differences between above indicators in experimental group and model group were statistically significant(P<0.01,P<0.05).Conclusion This study preliminarily established a methamphetamine-dependent cell model,and also found that rhynchophylline may regulate miR-375-3p/Elavl4 expression to antagonize methamphetamine addiction.
10.Study of Hedysarum polybotrys polysacchcaide regulating FXR-SHP pathway to improve glucose and lipid metabolism in liver tissue of diabetes rats
Lei ZHANG ; Sheng-Fang WAN ; Ya-Ling LI ; Qian-Kun LIANG ; Yi-Hong TIAN ; Xin-Xin MA ; Qian GUO
The Chinese Journal of Clinical Pharmacology 2024;40(17):2538-2542
Objective To investigate the effects of Astragalus polysaccharides(HPS)on farnitol X receptor(FXR)-small heterodimer chaperone receptor(SHP)signaling pathway and key proteins of glucose and lipid metabolism in diabetic rats.Methods Twelve male Wistar rats were randomly selected as blank group,and the remaining 60 rats were fed with a one-time intrabitoneal injection of streptozotocin(STZ,50 mg·kg-1)combined with a high-sugar and high-fat diet to replicate the diabetic rat model.The model rats were randomly divided into model group,positive control group(400 mg·kg-1·d-1 Bifidobacterium quadruple viable bacterial tablet suspension),experimental-H,-M,-L groups(200,100 and 50 mg·kg-1·d-1 HPS suspension),respectively.Blank group and model group were given equal volume of pure water once a day for 8 weeks.Blood glucose(Glu)was measured before and after gavage.Real-time fluorescent polymerase chain reaction(RT-PCR),Western blot were used to detect the mRNA and protein expression level of FXR,SHP,antiperoxisomal proliferator-activated receptor α(PPARα),antiphosphoenolpyruvate carboxylkinase(PEPCK),sterol regulatory receptor binding protein-1c(SREBP-1c),glucose 6 phosphatase(G6Pase).Results Glu levels in normal group,model group,positive control group and experimental-H group after treatment were(7.66±0.61),(29.25±1.64),(23.31±3.02),(19.31±5.13)mmol·L-1,respectively;the relative expression levels of FXR mRNA in liver tissues were 1.00±0.04,0.44±0.03,0.61±0.06,0.87±0.03,respectively;the relative expression levels of SHP mRNA were 1.00±0.04,0.40±0.01,0.67±0.01,0.67±0.02;the relative expression levels of G6Pase mRNA in liver tissues were 1.00±0.06,3.00±0.08,1.87±0.03,1.44±0.05,respectively;the relative expression levels of PEPCK mRNA in liver tissues were 1.00±0.04,1.88±0.03,1.31±0.02,1.23±0.04,respectively;the relative expression levels of SREBP-1c mRNA in liver tissues were 1.00±0.04,1.90±0.01,1.26±0.03,1.06±0.04;the relative expression levels of PPARα mRNA in liver tissues were 1.00±0.02,0.16±0.01,0.45±0.01,0.96±0.03,respectively.Compared with blank group,positive control group and experimental-H group,there were statistically significant differences in the above indexes between model group and blank group(all P<0.01).The protein expression trend of FXR,SHP,G6Pase,PEPCK,SREBP-1c,PPARα was consistent with mRNA expression.Conclusion HPS may regulate FXR-SHP signaling pathway in liver tissue,inhibit the expression of key proteins of glucose and lipid metabolism,promote lipid oxidation,improve Glu and protect liver tissue in diabetic rats.

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