Changes of plasma Rap1A levels in patients with in-stent restenosis after percutaneous coronary intervention and the underlying mechanisms / 中南大学学报(医学版)

Objective:Percutaneous coronary intervention(PCI)is one of the most important treatments for coronary artery disease(CAD).However,in-stent restenosis(ISR)after PCI is a serious complication without effective measures for prevention and treatment.This study aims to investigate the Ras-related protein 1A(Rap1A)level in ISR patients and in the tumor necrosis factor-α(TNF-α)-induced inflammatory injury model of human umbilical vein endothelial cells(HUVECs),to explore the role of Rap1A in regulating TNF-α-induced inflammation in HUVECs and to provide a new potential target for ISR prevention and treatment. Methods:A total of 60 CAD patients,who underwent PCI between December 2020 and July 2022 from the Department of Cardiovascular Medicine of Xiangya Hospital,Central South University,and re-examined coronary angiography(CAG)1 year after the operation,were included.After admission,27 patients were diagnosed with ISR and 33 patients were diagnosed with non-in-stent restenosis(non-ISR)according to the CAG.Clinical data were collected,and the plasma Rap1A level was determined by enzyme linked immunosorbent assay(ELISA).In cell experiments,an inflammatory injury model was established with TNF-α treatment(10 ng/mL,24 h)in HUVECs.The mRNA and protein expression levels of Rap1A,interlukin-6(IL-6),and vascular cell adhesion molecule-1(VCAM-1)were measured by real-time reverse transcription PCR and Western blotting.Small interfering RNA(siRNA)was used to explore the role of Rap1A in regulating TNF-α-induced inflammation in HUVECs. Results:Compared with the non-ISR patients,a higher proportion of ISR patients had a history of smoking(P=0.005)and diabetes(P=0.028),and higher levels of glycosylated hemoglobin(HbA1c)(P=0.012),low-density lipoprotein cholesterol(LDL-c)(P=0.014),and hypersensitive C-reactive protein(hs-CRP)(P=0.027).The remaining projects did not show significant differences(all P＞0.05).The plasma level of Rap1A in the ISR group was significantly higher than that in the non-ISR group[942.14(873.28 to 1 133.81)μg/mL vs 886.93(812.61 to 930.98)μg/mL;P=0.004].Diabetes,LDL-c,and Rap1A were risk factors for ISR by univariate logistic regression analysis(all P＜0.05).The mRNA and protein expression levels of inflammatory factors IL-6 and VCAM-1 were increased in HUVECs after 10 ng/mL TNF-α treatment for 24 h compared with the control group(all P＜0.05),while the mRNA and protein levels of Rap1A were increased(both P＜0.05).After inhibition of Rap1A in HUVECs,the mRNA and protein expression levels of IL-6 and VCAM-1 were significantly decreased(all P＜0.05). Conclusion:The plasma Rap1A level was significantly elevated in patients with ISR,suggesting that Rap1A may be a potential biomarker for predicting ISR.In the TNF-α-induced HUVECs inflammatory injury model,the expression level of Rap1A was increased.The level of TNF-α-induced endothelial cell inflammation was decreased after inhibition of Rap1A expression,suggesting that Rap1A may be a potential target for the treatment of endothelial cell inflammation in ISR.

Levels of plasma Quaking and cyclooxygenase-2 predict in-stent restenosis in patients with coronary artery disease after percutaneous coronary intervention / 中南大学学报(医学版)

OBJECTIVES@#Percutaneous coronary intervention (PCI) is one of the important methods for the treatment of coronary artery disease (CAD). In-sent restenosis (ISR) after PCI for patients suffered from CAD is considered to be an essential factor affecting long-term outcomes and prognosis of this disease. This study aims to investigate the correlation between plasma Quaking (QKI) and cyclooxygenase-2 (COX-2) levels and ISR in patients with CAD.@*METHODS@#A total of 218 consecutive CAD patients who underwent coronary angiography and coronary arterial stenting from September 2019 to September 2020 in the Department of Cardiology of Xiangya Hospital of Central South University were enrolled in this study, and 35 matched individuals from the physical examination center were served as a control group. After admission, clinical data of these 2 groups were collected. Plasma QKI and COX-2 levels were measured by enzyme linked immunosorbent assay (ELISA). Follow-up angiography was performed 12 months after PCI. CAD patients were divided into a NISR group (n=160) and an ISR group (n=58) according to the occurrence of ISR based on the coronary angiography. The clinical data, coronary angiography, and stent features between the NISR group and the ISR group were compared, and multivariate logistic regression was used to explore the factors influencing ISR. The occurrence of major adverse cardiovascular events (MACE) 1 year after operation was recorded. Fifty-eight patients with ISR were divided into an MACE group (n=24) and a non-MACE group (n=34), classified according to the occurrence of MACE, and the plasma levels of QKI and COX-2 were compared between the 2 groups. Receiver operating characteristic (ROC) curves were utilized to analyze the diagnostic value of plamsa levels of QKI and COX-2 for ISR and MACE occurrences in patients after PCI.@*RESULTS@#Compared with control group, plasma levels of QKI and COX-2 in the CAD group decreased significantly (all P<0.001). Compared with the NISR group, the plasma levels of QKI and COX-2 also decreased obviously in the ISR group (all P<0.001), while the levels of high sensitivity C-reactive protein (hs-CRP) and glycosylated hemoglobin (HbAlc) significantly increased (all P<0.001). The level of COX-2 was negatively correlated with hs-CRP (r=-0.385, P=0.003). Multivariate logistic regression analysis showed that high level of plasma QKI and COX-2 were protective factors for in-stent restenosis after PCI, while hs-CRP was a risk factor. ROC curve analysis showed that the sensitivity and specificity of plasma QKI for evaluating the predictive value of ISR were 77.5% and 66.5%, respectively, and the sensitivity and specificity of plasma COX-2 for evaluating the predictive value of ISR were 80.0% and 70.7%, respectively. The sensitivity and specificity of plasma QKI combined with COX-2 for evaluating the predictive value of ISR were 81.3% and 74.1%, respectively. The sensitivity and specificity of plasma QKI for evaluating the prognosis of ISR were 75.0% and 64.7%, respectively. The sensitivity and specificity of plasma COX-2 for evaluating the prognosis of ISR were 75.0% and 70.6%, respectively. The sensitivity and specificity of plasma QKI combined with COX-2 for prognostic evaluation of ISR were 81.7% and 79.4%, respectively. The sensitivity and specificity of plasma COX-2 combined with QKI for evaluating ISR and MACE occurrences in patients after PCI were better than those of COX-2 or QKI alone (P<0.001).@*CONCLUSIONS@#High level of plasma QKI and COX-2 might be a protective factor for ISR, which can also predict ISR patient's prognosis.

Extraction and Content Determination of Polysaccharide in Mongolian Medicine Vicia amoena / 中国药房

OBJECTIVE:To optimize the decolorization condition for polysaccharide extract of Mongolian medicine Vicia amoena,and to establish the method for its extraction and content determination. METHODS:The water extract-alcohol precipita-tion was used to extract polysaccharide from Mongolian medicine V. amoena. Using decolorization rate as index,orthogonal test was designed to investigate the effects of the dosage of activated carbon,decolorization temperature,decolorization time on the de-colorization of polysaccharide,so as to optimize the conditions for the decoloration of polysaccharide. Using glucose as control, phenol sulfuric acid method was adopted,and the content of polysaccharide in crude polysaccharide was determined by UV spectro-photometry at 490 nm. RESULTS:The optimal decoloration condition was as follows as actived carbon of 3%,decoloration time of 40 min,decoloration temperature of 60 ℃. On this basis,the average decolorization rate reached 19.77%(RSD=1.85%,n=3) by the verification test of the decoloration. The average extraction yield for the crude polysaccharide was 4.56%(RSD=2.38%,n=3),of which the polysaccharide content was 1.98%(RSD=2.18%,n=4). CONCLUSIONS:This experiment has relatively good reproducibility in polysaccharide yield;established method for content determination of polysaccharide is stable and feasible.

Expression of Monocyte Chemotactic Protein-1 and Its Receptor in Sudden Coro-nary Death / 法医学杂志

Objective To investigate the expression of m onocyte chem otactic protein-1 (MCP-1) and its receptor CCchem okine receptor-2 (CCR-2) in coronary atherosclerosis plaques betw een sudden coro-nary death (SCD ) and non-SCD. Methods The expression levels of MCP-1 and CCR-2 in SCD group, coronary atherosclerosis group (non-SCD), control group (norm al coronary artery) w ere detected by im-m unohistochem istry. Results Positive rates of MCP-1 am ong the three groups w ere 78%, 47%, and 0%, respectively, w ith significant expressing differences betw een each tw o groups (P0.05). Conclusion O verexpression of MCP-1 and CCR-2 in coronary atherosclerotic plaques is closely correlated w ith SCD .

Metabonomics Study on Luteolin Intervention of Breast Cancer MCF-7 Cells / 分析化学

The metabolic profiles of control and MCF-7 cells treated with luteolin were analyzed separately using gas chromatography/mass spectrometry ( GC/MS ) to study the mechanism of the luteolin treatment on MCF-7 cells. Cell viability assays showed that luteolin had inhibition effect on MCF-7 cells. Partial least square discriminant analysis ( OPLS-DA) was used to process the metabolic data. Since cells in phase of S were increased significantly, we speculated that luteolin had a blocking effect on pentose phosphate pathway of MCF-7 cells, which contributed to its inhibition effect on proliferation of MCF-7 cells.