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.Health impact assessment of the development of employment and social security in Deqing County, Zhejiang Province
Shiyu HAN ; Bingbing ZHU ; Yiming ZHANG ; Yuting YANG ; Chaowei FU
Shanghai Journal of Preventive Medicine 2024;36(3):274-279
ObjectiveTo identify the possible health impact of the 14th five⁃year plan for the development of employment and social security in Deqing County and propose improvement measures through health impact assessment. MethodsBased on the data of Deqing County, stakeholder interviews and Delphi Consultation Method, this study described the current status of employment and social security and analyzed the potential health impacts of implementing the 14th five⁃year plan for the development of employment and social security in Deqing County. ResultsThrough a quick assessment process, the results showed that the implementation of the plan would bring mixed health impacts. Positive impacts included enhanced social security capacity, improved health levels of low-income populations and families, increased convenience of medical treatment, and improved efficiency of health services. Negative impacts included reduced accessibility of digital services for the elderly, increased gap in benefits for retirees, increased risk of discrimination against disabled individuals, increased risks of layoffs and unemployment for vulnerable groups, and increased employment instability for middle-aged and elderly populations. ConclusionThe 14th five⁃year plan for the development of employment and social security in Deqing County will bring a series of positive health impacts, but negative health impacts also warrant attention.
3.Motor cortex stimulation for the treatment of trigeminal neuralgia after brainstem infarction:A case report
Journal of Central South University(Medical Sciences) 2024;49(1):54-57
Secondary trigeminal neuralgia after brainstem infarction is rare and rarely reported.A patient with secondary trigeminal neuralgia after brainstem infarction was admitted to the Department of Neurosurgery,Xiangya Hospital,Central South University.The patient was a 44 years old male who underwent motor cortex stimulation treatment after admission.The effect was satisfactory in the first week after surgery,but the effect was not satisfactory after one week.This disease is relatively rare and the choice of clinical treatment still requires long-term observation.
4.Diagnostic Efficacy and Influencing Factors of Ultrasonography Combined with Contrast-Enhanced Ultrasound in Breast Lesions
Naiqin FU ; Junkang LI ; Ying JIANG ; Shiyu LI ; Ruilan NIU ; Zhili WANG
Chinese Journal of Medical Imaging 2024;32(1):67-72
Purpose To explore the diagnostic efficiency of ultrasound(US)combined with contrast-enhanced ultrasound(CEUS)in breast lesions and to analyze the related factors affecting the diagnostic accuracy.Materials and Methods From January 2022 to February 2023,the clinical data and ultrasound images of 784 patients who underwent breast US and CEUS examination with definite pathological results were retrospectively collected in the First Medical Center of the PLA General Hospital.The diagnostic efficacy of US combined with CEUS in benign and malignant breast lesions was analyzed,respectively.The independent risk factors for diagnostic errors were analyzed via Logistic regression.Results The sensitivity,specificity and accuracy of US combined with CEUS in the diagnosis of benign and malignant breast lesions were 89.2%,84.4%and 88.7%,respectively.The area under the receiver operating characteristic curve was 0.932.The results of multivariate Logistic regression analysis showed that the diagnosis error rate increased when the lesions were non-mass type(odds ratio,OR=1.927,P=0.047),complex cystic and solid(OR=3.729,P=0.000),and high-enhanced CEUS(OR=1.937,P=0.023),while the diagnosis error rate decreased when the lesions were large(OR=0.688,P=0.004)and with US-detect suspicious lymph node(OR=0.143,P=0.011).Conclusion When the breast lesions are non-mass type,complex cystic and solid lesions and hyper-enhancement,the diagnosis error rate of US combined with CEUS increased.It is necessary to further explore the enhancement patterns of different lesions.
5.Tougu Xiaotong Capsule alleviates cartilage degeneration in mice with knee osteoarthritis by modulating Nav1.7
Changlong FU ; Yanming LIN ; Shujie LAN ; Yue CHEN ; Chao LI ; Shiyu LU ; Qing LIN
Journal of Southern Medical University 2024;44(11):2074-2081
Objective To investigate the mechanism by which Tougu Xiaotong Capsule(TGXTC)alleviates chondrocyte degeneration in knee osteoarthritis(KOA).Methods Thirty 2-month-old C57BL/6 mouse models of KOA established using the Hulth method were randomized into model group,TGXTC group,and diclofenac sodium group and received treatment with saline,TGXTC(368 mg/kg),and diclofenac sodium(10 mg/kg)by gavage,respectively,with another 10 untreated mice as the blank control group.All interventions were administered 6 times a week for 4 weeks.After the treatments,structural changes in the cartilage tissue were observed with morphological staining,and Nav1.7 mRNA expression and the protein expression levels of Nav1.7,MMP-3,ADAMTS-5,and COX-2 were detected using RT-qPCR and Western blotting.Fluorescence in situ hybridization(FISH)was used to detect Nav1.7 expression in the chondrocytes.In cultured KOA chondrocytes,the effect of TGXTC and lentivirus-mediated Nav1.7 knockdown on MMP-3,MMP-13,ADAMTS-4,ADAMTS-5,and COX-2 protein expressions were assessed with Western blotting.Results In KOA mice treatments with TGXTC and diclofenac sodium both significantly alleviated structural damage of the cartilage layer,reduced Nav1.7 protein and mRNA expressions and lowered the expressions of MMP-3,ADAMTS-5,and COX-2 proteins in the cartilage tissues.FISH results indicated that TGXTC treatment significantly reduced IL-1β-induced Nav1.7 expression in the chondrocytes.In Nav1.7 knockdown experiment,Nav1.7 levels were significantly lower in IL-1β+sh-Nav1.7 group than in IL-1β group,and also lower in IL-1β+TGXTC group than in IL-1β+sh-Nav1.7+TGXTC group.TGXTC treatment significantly inhibited IL-1β-induced elevation of MMP-3,MMP-13,ADAMTS-4,ADAMTS-5 and COX-2 protein expressions in the chondrocytes,but its effects were strongly weakened by Nav1.7 knockdown.Conclusion TGXTC alleviates extracellular matrix metabolic disorder in KOA chondrocytes by regulating Nav1.7,thereby mitigating chondrocyte degeneration in KOA mice.
6.Research progress in programmed cell death protein ligand-1 targeting peptide-based radionuclide-labeled molecular probes
Shiyu ZHU ; Beibei LIANG ; Jiayu FU ; Jianguo LIN ; Ling QIU
Journal of Xinxiang Medical College 2024;41(5):491-496
Programmed cell death protein ligand-1(PD-L1)is an important immune checkpoint molecule that plays an important role in regulating the body's immune response.Several clinical studies have shown that the expression level of PD-L1 in tumors is closely related to the efficacy of immune checkpoint inhibitors.Due to the spatial and temporal heterogeneity of tumors,immunohistochemical methods commonly used in clinical practice cannot accurately and comprehensively reflect the ex-pression level of PD-L1 in patients.Given that nuclear-medicine molecular imaging technology can noninvasively,real-time,dy-namically and visually monitor the expression level of PD-L1 in vivo at the molecular level,this article mainly focuses on the re-search of peptide-based radiolabeled molecular probes targeting PD-L1,with the aim of providing guidance for the search of no-vel peptide molecular probes for immunoimaging as well as for the screening of immunotherapy-suitable patients and evaluation of therapeutic efficacy,and other clinical applications.
7.Tougu Xiaotong Capsule alleviates cartilage degeneration in mice with knee osteoarthritis by modulating Nav1.7
Changlong FU ; Yanming LIN ; Shujie LAN ; Yue CHEN ; Chao LI ; Shiyu LU ; Qing LIN
Journal of Southern Medical University 2024;44(11):2074-2081
Objective To investigate the mechanism by which Tougu Xiaotong Capsule(TGXTC)alleviates chondrocyte degeneration in knee osteoarthritis(KOA).Methods Thirty 2-month-old C57BL/6 mouse models of KOA established using the Hulth method were randomized into model group,TGXTC group,and diclofenac sodium group and received treatment with saline,TGXTC(368 mg/kg),and diclofenac sodium(10 mg/kg)by gavage,respectively,with another 10 untreated mice as the blank control group.All interventions were administered 6 times a week for 4 weeks.After the treatments,structural changes in the cartilage tissue were observed with morphological staining,and Nav1.7 mRNA expression and the protein expression levels of Nav1.7,MMP-3,ADAMTS-5,and COX-2 were detected using RT-qPCR and Western blotting.Fluorescence in situ hybridization(FISH)was used to detect Nav1.7 expression in the chondrocytes.In cultured KOA chondrocytes,the effect of TGXTC and lentivirus-mediated Nav1.7 knockdown on MMP-3,MMP-13,ADAMTS-4,ADAMTS-5,and COX-2 protein expressions were assessed with Western blotting.Results In KOA mice treatments with TGXTC and diclofenac sodium both significantly alleviated structural damage of the cartilage layer,reduced Nav1.7 protein and mRNA expressions and lowered the expressions of MMP-3,ADAMTS-5,and COX-2 proteins in the cartilage tissues.FISH results indicated that TGXTC treatment significantly reduced IL-1β-induced Nav1.7 expression in the chondrocytes.In Nav1.7 knockdown experiment,Nav1.7 levels were significantly lower in IL-1β+sh-Nav1.7 group than in IL-1β group,and also lower in IL-1β+TGXTC group than in IL-1β+sh-Nav1.7+TGXTC group.TGXTC treatment significantly inhibited IL-1β-induced elevation of MMP-3,MMP-13,ADAMTS-4,ADAMTS-5 and COX-2 protein expressions in the chondrocytes,but its effects were strongly weakened by Nav1.7 knockdown.Conclusion TGXTC alleviates extracellular matrix metabolic disorder in KOA chondrocytes by regulating Nav1.7,thereby mitigating chondrocyte degeneration in KOA mice.
8.Exploration of the relationship between the storage time of leukodepleted red blood cell and transfusion adverse reactions
Liu HE ; Jian LIU ; Gang WU ; En WANG ; Fayan YI ; Xingshun TAN ; Shiyu ZHU ; Rui YU ; Guanghui LU ; Yan LIU ; Mei ZHAI ; Qing XIANG ; Ping LIU ; Yanhua LIAO ; Zhizhen FU ; Maolin LI ; Rong HUANG
Chinese Journal of Blood Transfusion 2023;36(10):889-891
【Objective】 To explore the relationship between the storage time of leukodepleted red blood cells and transfusion adverse reactions by analyzing the occurrence of transfusion adverse reactions of patients after leukodepleted red blood cells transfusion from four hospitals. 【Methods】 By using the electronic medical record management system, the collection and transfusion dates of leukodepleted red blood cells from four hospitals in Enshi Prefecture from 2018 to 2022, as well as the information on transfusion adverse reactions, were retrieved. 【Results】 From 2018 to 2022, a total of 697 61 bags of leukodepleted red blood cells were transfused in four hospitals, resulting in 166 cases of transfusion adverse reactions, among which 93 were allergic reactions, 63 were non hemolytic febrile reactions, and 10 were others, with a total incidence rate of transfusion adverse reactions at 0.24%. The average storage time of leukodepleted red blood cells with and without transfusion adverse reactions was (20.25±6.31) and (19.88±5.50) days, respectively. With a storage time of 7 days as the threshold, the incidence of transfusion adverse reactions was the lowest for a storage time of 15~21 days. The incidence of transfusion adverse reactions of leukodepleted red blood cells in two groups (with storage days ≤21 days and >21 days) was not statistically significant(P>0.05). 【Conclusion】 Allergic reactions were the main type of transfusion adverse reaction caused by leukodepleted red blood cells, and the incidence of transfusion adverse reactions decreased and then increased with the prolongation of the storage time of leukodepleted red blood cells. There was no significant difference in the incidence of transfusion adverse reactions with leukodepleted red blood cells stored for ≤ 21 days and >21 days.
9.Development of a radiolabeled site-specific single-domain antibody positron emission tomography probe for monitoring PD-L1 expression in cancer
Yinfei CHEN ; Shiyu ZHU ; Jiayu FU ; Jianguo LIN ; Yan SUN ; Gaochao LV ; Minhao XIE ; Tao XU ; Ling QIU
Journal of Pharmaceutical Analysis 2022;12(6):869-878
Despite advances in immunotherapy for the treatment of cancers,not all patients can benefit from programmed cell death ligand 1(PD-L1)immune checkpoint blockade therapy.Anti-PD-L1 therapeutic effects reportedly correlate with the PD-L1 expression level;hence,accurate detection of PD-L1 expression can guide immunotherapy to achieve better therapeutic effects.Therefore,based on the high affinity antibody Nb109,a new site-specifically radiolabeled tracer,68Ga-NODA-cysteine,aspartic acid,and valine(CDV)-Nb109,was designed and synthesized to accurately monitor PD-L1 expression.The tracer 68Ga-NODA-CDV-Nb109 was obtained using a site-specific conjugation strategy with a radiochemical yield of about 95%and radiochemical purity of 97%.It showed high affinity for PD-L1 with a dissociation constant of 12.34±1.65 nM.Both the cell uptake assay and positron emission tomography(PET)imaging revealed higher tracer uptake in PD-L1-positive A375-hPD-L1 and U87 tumor cells than in PD-L1-negative A375 tumor cells.Meanwhile,dynamic PET imaging of a NC1-H1299 xenograft indicated that doxorubicin could upregulate PD-L1 expression,allowing timely interventional immunotherapy.In conclusion,this tracer could sensitively and dynamically monitor changes in PD-L1 expression levels in different cancers and help screen patients who can benefit from anti-PD-L1 immunotherapy.
10.Comparison of the Contents of 14 Inorganic Elements in Cyperus rotundus from Different Origins Based on Stoichiometry
Na XU ; Shiyu WANG ; Wenbing LI ; Chaomei FU ; Hulan CHEN ; Junrong LU
China Pharmacy 2021;32(3):289-294
OBJECTIVE:To establish the method for the content determination of inorganic elements in Cyperus rotundus ,and to compare the contents of 14 kinds of inorganic elements in C. rotundus from different producing areas ,and to provide theoretical basis for its quality control and high quality resources development . METHODS :The samples were processed by microwave digestion,and ICP-MS method was used to determine the contents of Na ,Mg,K,Ca,Mn,Fe,Ni,Cu,Zn,As,Se,Sr,Cd and Pb. SPSS 23.0 software were used for principal component analysis (PCA)and cluster analysis. RESULTS :The average contents of above 14 kinds of inorganic elements in C. rotundus from different producing areas were 168.62,753.71, 6 938.33,24.31,14.69,197.77,0.60,2.43,26.89,0.21,0.06,5.64,0.05,0.32 mg/kg,respectively. The results of PCA showed that the cumulative variance contribution rate of the first four principal components was 86.203%,which could reflect most of the information of the original data. C. rotundus from Shandong ,Jiangxi,Shanxi,Hubei and Yunnan ranked the top five places in terms of comprehensive score of inorganic element contents. The results of cluster analysis showed that the samples from 9 producing areas were clustered into 5 categories,showing the characteristics of clustering by producing area. From the perspective of inorganic elements ,the quality of C. rotundus from East China ,Central China ,North China and Southwest China was better than that from South China. CONCLUSIONS :Essential trace elements like Na ,Mg,K,Ca,Mn,Fe,Cu,Zn,Sr are rich in C. rotundus,and there are small amounts of Ni ,As,Se,Cd,Pb elements in it. The contents difference of inorganic elements in C. rotundus from different origins may related to the geographical area it belongs to.


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