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.Progress in preclinical studies of xenogeneic lung transplantation and single-center technical experience
Xiaoting TAO ; Xinzhong NING ; Yong LIU ; Guimei ZHANG ; He XIAO ; Shiyu LIN ; Zizi ZHOU ; Taiyun WEI ; Chunxiao HU ; Hongjiang WEI ; Kun QIAO
Organ Transplantation 2025;16(6):874-880
Lung transplantation is the ultimate therapeutic option for end-stage pulmonary diseases such as interstitial pneumonia, chronic obstructive pulmonary disease and pneumoconiosis. Currently, the shortage of allogeneic lung donors significantly limits the opportunity for end-stage lung disease patients to receive lung transplantation. In recent years, with the rapid development of biomedical engineering technologies, especially the major breakthroughs in genetic modification and cloning, xenogeneic lung transplantation has shown important potential for clinical translation. Among them, genetically modified pigs have become the most promising xenogeneic lung source due to the close similarity of organ size and physiological characteristics to humans, and the ability to perform targeted gene knockouts (such as α-Gal antigen knockout) to reduce the occurrence of hyperacute rejection. This article focuses on the research progress of porcine xenogeneic lung transplantation, systematically reviews the latest achievements and challenges in animal experiments and human trials, and introduces the technical experience accumulated by Shenzhen Third People's Hospital in the porcine-to-monkey xenogeneic lung transplantation model, in the hope of providing practical references for future research in this field.
3.Application of PDCA cycle in DRG payment for oncological diseases
Ke CAO ; Bonian CHEN ; Shiyu LIN ; Yue YI
Modern Hospital 2024;24(2):263-266,270
Objective This study aims to investigate the application of the Plan-Do-Check-Act(PDCA)cycle in the Di-agnosis Related Group(DRG)payment system for oncological diseases.Methods We analyzed the factors influencing DRG pay-ment and incorporated the PDCA Cycle in the oncological diseases at a tertiary hospital in Tianjin.The baseline data of cases partic-ipating in DRG payment from April 2022 to September 2022 were compared with the settlement data of cases participating in DRG payment from October 2022 to March 2023.SPSS 20.0 was used to evaluate the impact of data quality in medical record documenta-tion on DRG admission rate,average hospitalization costs,average length of stay,Case Mix Index(CMI)value,and DRG settle-ment rate,so as to assess the effectiveness of the PDCA cycle in DRG payment within the oncological diseases at the hospital.Re-sults Following the PDCA cycle,the DRG admission rate increased from 84.03%to 89.98%,and the cases ineligible for inclu-sion decreased by 22.78%due to mismatched main diagnosis and procedures."Violations of the reporting and coding principles that do not require reporting"and"omission of primary surgical procedure codes"were no longer observed as reasons for failed DRG inclusion.Ambiguous cases with both average hospitalization costs and average length of stay higher than those of normal inclu-sion cases,leading to an increase in the average hospitalization cost from 22 496.56 yuan to 24714.92 yuan,and the average length of stay increased from 7.50 days to 8.13 days.The CMI value increased from 0.96 to 1.08,and the DRG settlement rate increased from 107.93%to 130.67%.Conclusion The PDCA cycle can effectively enhance the quality of medical record documentation,leading to improved quality in medical insurance settlement lists and DRG admission rates.It can help identify operational issues within the de-partment and promote the smooth implementation of DRG payment reform in the oncological department.
4.Risk factors of abnormal urinary albumin/creatinine ratio in people with obesity
Zhe CAO ; Tongyue YANG ; Shiyu LIU ; Mengxing PAN ; Xuyang GONG ; Qianshuai LI ; Jiao WANG ; Lin ZHAO ; Guijun QIN ; Yanyan ZHAO
Chinese Journal of Endocrinology and Metabolism 2024;40(3):186-191
Objective:To explore the clinical characteristics and risk factors of abnormal urinary albumin/creatinine ratio(UACR) in obese population.Methods:Baseline data from 2011 to 2012 in Henan Sub-center of"Risk Evaluation of cAncers in Chinese diabeTic Individuals: A lONgitudinal(REACTION) study"were utilized and those of body mass index≥28 kg/m 2 were screened. The patients were divided into UACR normal group and UACR abnormal group(101 pairs) upon being matched on a 1∶1 basis by age and gender. Multivariate logistic regression analysis, receiver operating characteristic(ROC) curve, and restricted cubic spline(RCS)analysis were performed to explore the risk factors for abnormal UACR. Results:Compared with the normal UACR group, the UACR abnormal group had a higher number of alcohol consumers, a higher prevalence of hypertension, elevated systolic blood pressure, and triglyceride(all P<0.05). Multivariate logistic regression analysis showed that alcohol consumption( P=0.008), systolic blood pressure( P<0.001), triglyceride( P=0.049), and homeostasis model assessment for insulin resistance(HOMA-IR, P=0.033) were independent risk factors for abnormal UACR in obese people. The ROC curve analysis indicated that systolic blood pressure had the strongest diagnostic performance as a single factor(ROC curve area=0.801), and there was no significant difference in diagnostic performance compared to multiple factors combination. RCS analysis results showed that the probability of abnormal UACR increased monotonically with the increase of systolic blood pressure when the systolic blood pressure was between 130 and 158 mmHg(1 mmHg=0.133 kPa). When systolic blood pressure was not in the interval, the probability of abnormal UACR did not change significantly. The results of regression analysis of triglyceride subgroup showed that when triglyceride level was greater than or equal to 5.6 mmol/L, the risk of abnormal UACR level was significantly increased( P=0.029). Conclusion:Systolic blood pressure, triglyceride, HOMA-IR, and alcohol drinking history are independent risk factors for abnormal UACR in obese people. When systolic blood pressure is≥130 mmHg or triglyceride is≥5.6 mmol/L, the risk of abnormal UACR is significantly increased.
5.Effects of Point Ligation Therapy Combined with Acupoint Catgut Embedment on Facial Nerve Electromyo-graphy and Serum Neurotrophic Factors for Patients with Refractory Facial Paralysis
Hongzhu LI ; Guoan LAI ; Shiyu LIN ; Xiuyi MA ; Guangxian CHEN ; Yuemei LI ; Shuxin WANG
Journal of Traditional Chinese Medicine 2024;65(15):1578-1585
ObjectiveTo observe the clinical effectiveness of point ligation therapy combined with catgut embedment in acupoint for patients with refractory facial paralysis, and to explore the possible mechanisms from the perspective of neurotrophic factors. MethodsTotally 168 patients with intractable facial paralysis were randomly divided into point ligation plus catgut embedment group and electroacupuncture group, with 84 cases in each group. Both groups took methylcobalamin tablets orally, on the basis of which the point ligation plus catgut embedment group gave point ligation therapy at Quanliao (SI 18), Dicang (ST 4), Jiache (ST 6), and catgut embedment in the local acupoints on the affected side combined with the distal acupoints, and the point ligation therapy combined with catgut embedment in acupoint was performed once every 30 days for three treatments; the electroacupuncture group took the same acupoints and gave electroacupuncture, with sparse and dense waves, once every other day. Both groups were treated for 90 days. Before treatment and after 30, 60, 90 and 120 days (follow-up), patients of both groups were observed for House-Brackmann Facial Nerve Grading (HB) and modified Portmann score; facial nerve electromyography was performed before treatment and after 90 days of treatment to record the wave amplitude and latency of facial nerve motor conduction, and serum brain-derived nerve growth factor (BDNF), nerve growth factor (NGF), fibroblast growth factor (FGF2) levels were determined. The clinical effectiveness of both groups after 90 days of treatment was compared, and the adverse events occurred in the course of treatment in all patients were recorded. ResultsAt 30 days, 60 days, 90 days and follow-up visits, the HB grading of patients in both groups improved compared with that before treatment (P<0.05); the distribution of the number of patients with HB grading in the point ligation plus catgut embedment group was superior to that in the electroacupuncture group after 60 days and 90 days of treatment and at follow-up visits (P<0.05). The maximal amplitude of the facial nerve electromyography in both groups after 90 days of treatment was large, and the latency was shorter when compared with those before treatment (P<0.05), and the maximum wave amplitude of the point ligation plus catgut embedment group was higher than that of the electro-acupuncture group, and the latency period was shorter than that of the electro-acupuncture group (P<0.05). The modified Portmann scores of the both groups of patients after 30, 60 and 90 days of treatment and follow-up were higher than those before treatment, and the scores of the point ligation plus catgut embedment group were higher than those of the electroacupuncture group (P<0.05). Serum BDNF, NGF and FGF2 levels in both groups after 90 days of treatment were higher than before, and the point ligation plus catgut embedment group was significantly higher than the electroacupuncture group (P<0.05). The total clinical effective rate of the point ligation plus catgut embedment group (92.68%) was higher than that of the electroacupuncture group (81.01%, P<0.05). Adverse reactions occurred in both groups, mainly including local haematoma or subcutaneous bleeding, allergic reaction, severe pain, and needle fainting, all of which disappeared after symptomatic treatment. ConclusionThe point ligation therapy combined with catgut embedment in acupoint for patients with refractory facial paralysis are effective. The therapy improve the facial nerve function, and its mechanism of action may be related to increasing the level of serum neurotrophic factor and thus promoting facial nerve repairment.
6.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.
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.Predicting Acute Mountain Sickness Using Regional Sea-Level Cerebral Blood Flow
Zhang HAO ; Feng JIE ; Zhang SHIYU ; Liu WENJIA ; Ma LIN
Biomedical and Environmental Sciences 2024;37(8):887-896
Objective To investigate the role of sea-level cerebral blood flow(CBF)in predicting acute mountain sickness(AMS)using three-dimensional pseudo-continuous arterial spin labeling(3D-pCASL). Methods Forty-eight healthy volunteers reached an altitude of 3,650 m by air after undergoing a head magnetic resonance imaging(MRI)including 3D-pCASL at sea level.The CBF values of the bilateral anterior cerebral artery(ACA),middle cerebral artery(MCA),posterior cerebral artery(PCA),and posterior inferior cerebellar artery(PICA)territories and the laterality index(LI)of CBF were compared between the AMS and non-AMS groups.Statistical analyses were performed to determine the relationship between CBF and AMS,and the predictive performance was assessed using receiver operating characteristic(ROC)curves. Results The mean cortical CBF in women(81.65±2.69 mL/100 g/min)was higher than that in men(74.35±2.12 mL/100 g/min)(P<0.05).In men,the cortical CBF values in the bilateral ACA,PCA,PICA,and right MCA were higher in patients with AMS than in those without.Cortical CBF in the right PCA best predicted AMS(AUC=0.818).In women,the LI of CBF in the ACA was different between the AMS and non-AMS groups and predicted AMS with an AUC of 0.753. Conclusion Although the mechanism and prediction of AMS are quite complicated,higher cortical CBF at sea level,especially the CBF of the posterior circulatory system,may be used for prediction in male volunteers using non-invasive 3D-pCASL.
9.Optimization of DNA purification nano beads specifically for FFPE samples
Lin XIAO ; Xiaofang LÜ ; Shiyu CAO ; Dan JIANG ; Linyong SUN ; Kun LI ; Feng YE
Chinese Journal of Clinical and Experimental Pathology 2024;40(7):725-730
Purpose Focusing on the characteristics of for-malin fixed paraffin embedded(FFPE)samples,explored nano-magnetic bead nucleic acid extraction solutions with higher qual-ity/yield and continued to improve molecular pathology technolo-gy.Methods Alternative magnetic beads were synthesised in four major categories and 15 sub-categories and we screened to obtain high-quality/yield magnetic beads centred on FFPE samples.Simulated conventional tissues,simulated coarse needle punctures(liver),and simulated fiberoptic bronchoscopy sam-ples(lungs)were sectioned with the same number of serial slices in tubes.The nucleic acids of slices were extracted using the best magnetic beads screened in this study and common com-mercially available kits,and then perform comparison and purifi-cation quality parameters such as total amount and fragment size.The downstream applications of nucleic acids were validated by PCR and Sanger sequencing.Results Screening all homemade nanomagnetic beads centered on the DNA of FFPE samples,the total recoveries of the best performance nanomagnetic beads were obtained to be 58.5%±1.58%,and the total recoveries of five commercially available commercial magnetic beads and three do-mestic kit magnetic beads ranged from 18.68%to 40.71%.The total amount of DNA(ng)extracted from the same amount of tis-sue(serial slices),the nucleic acid yield of this study in simu-lated conventional tissues,simulated coarse needle punctures,and simulated fiberoptic bronchoscopy samples were increased by 39.49%-181.72%compared with those of the commercially a-vailable kits(P<0.05).The total amount of extracted nucleic acid from simulated fiberoptic bronchoscopy tissue sections can be more than 100 ng for 1 slice(4 μm)and more than 400 ng for 5 slices.Conclusion The DNA purification nanomagnetic beads screened with DNA from FFPE samples have a significant enhancement comparing to the existing commercial bead proto-cols,and provide space for quality assurance,automated testing,and program expansion for clinical molecular pathology testing.
10.Application of PDSA cycle in addressing the problem of decomposed hospitalization for stroke disease under the DRG payment model
Ke CAO ; Bonian CHEN ; Shiyu LIN ; Yue YI
Modern Hospital 2024;24(7):1070-1074
Objective To explore the application value of the Plan-Do-Study-Act(PDSA)cycle in reducing the occur-rence of decomposed hospitalization for stroke disease under the Diagnosis Related Group(DRG)payment model and validate the effectiveness of strategies to reduce decomposed hospitalization.Methods Taking stroke cases participating in DRG payment from January 2022 to December 2022 in a tertiary hospital in Tianjin as the baseline,the criteria for decomposed hospitalization were determined based on the"Implementation Rules for DRG/DIP Network Audit(Trial)"issued by the Tianjin Medical Insur-ance Fund Settlement Center.A PDSA cycle plan was developed according to the document content,and the PDSA cycle was im-plemented from January to December 2023.The relationship between the number of decomposed hospitalizations before and after the PDSA cycle was analyzed using SPSS 20.0 to determine the actual application effect.Results After the PDSA cycle,the number of decomposed hospitalizations decreased from1268 cases to26 cases,a total decrease of 97.95%.The number of cases in each target subgroup and the frequency of factors leading to decomposed hospitalization all showed a significant decrease.Con-clusion The PDSA cycle plays a significant role in addressing the problem of decomposed hospitalization for stroke disease under the DRG payment model.It can effectively reduce the occurrence of decomposed hospitalization,improve the quality of medical insurance settlement lists and medical records,and standardize clinical diagnosis and treatment behavior.


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