1.Analysis on Hemostatic Active Components in Moutan Cortex Carbonisata Based on Spectrum-effect Relationship
Qingguang LIANG ; Xiguang LIN ; Jiang MENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):183-190
ObjectiveTo identify the primary hemostatic active components in Moutan Cortex Carbonisata(MCC) based on the spectrum-effect relationship between the fingerprint and hemostatic efficacy, thereby providing a basis for characterizing its active constituents. MethodsUltra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Orbitrap MS/MS) was employed to establish the fingerprint profiles of 16 batches of MCC aqueous extracts and identify the common peaks. Activated partial thromboplastin time(APTT), an in vitro coagulation activity indicator, was measured for the 16 batches of samples using a semi-automated coagulometer. Grey relational analysis(GRA), Pearson correlation analysis, and partial least squares regression(PLSR) were comprehensively applied to screen potential hemostatic active components. For the identified active components, multi-dimensional pharmacological validation was conducted through in vitro coagulation assays measuring APTT, prothrombin time(PT), and thrombin time(TT), evaluation of hemostasis rate using a zebrafish cerebral hemorrhage model, and real-time quantitative polymerase chain reaction(Real-time PCR) detection of coagulation factor X(FⅩ) mRNA expression level. ResultsThe UPLC fingerprint of the aqueous extract of MCC was successfully established, identifying 12 common peaks. Among these, 9 chemical components were subsequently characterized using UPLC-Q-Orbitrap MS. Comprehensive application of GRA, Pearson correlation analysis, and PLSR analysis identified 5-hydroxymethylfurfural(5-HMF), gallic acid, 1-O-galloylglucose, and p-hydroxybenzoic acid as key hemostatic active constituents in MCC. In vitro coagulation assays confirmed that all four active components significantly shortened APTT and PT(P<0.05, P<0.01). The zebrafish cerebral hemorrhage model further validated their in vivo hemostatic efficacy, with each component significantly reducing hemorrhage area(P<0.05, P<0.01), yielding hemostasis rates of 31.20% for 5-HMF, 68.85% for gallic acid, 45.45% for 1-O-galloylglucose, and 45.60% for p-hydroxybenzoic acid, and demonstrating overall concentration-dependent effects. Real-time PCR analysis demonstrated that all active components significantly upregulated FⅩ mRNA expression(P<0.05, P<0.01), synergistically enhancing hemostasis. ConclusionBy integrating spectrum-effect relationship analysis and multi-dimensional efficacy validation, this study identified four hemostatic constituents from MCC, providing a scientific basis for elucidating its hemostatic material basis.
2.The change and correlation of glycogen phosphorylase isoenzyme BB in neonates with asphyxia complicated with myocardial injury
Qinghua MAO ; Lixing LIN ; Zhiling ZHANG ; Caixia AN ; Xiguang KANG
Journal of Clinical Pediatrics 2010;(3):226-230
Objective To study the change of glycogen phosphorylase isoenzyme BB(GPBB)in neonates with asphyxia complicated with myocardial injury and its correlation with various perinatal factors. Methods Sixty-four neonates with asphyxia(including 39 mild asphyxia and 25 severe asphyxiated neonates,30 neonates with and 34 without myocardial injures)were enrolled and 25 healthy neonates were studied as control. The plasma levels of GPBB were measured by enzyme-linked immunosorbent assay(ELISA). Myocardial enzymes,cardiac troponin I,electrocardiogram,chest X-ray were performed simultaneously. Results The plasma GPBB levels were significantly higher in neonates with myocardial injury(13.84,7.57 ng/ml)than those without myocardial injury(4.97,3.24 ng/ml)and control group(4.95,1.99 ng/ml)( P < 0.01). The sensitivities of GPBB,cTnI and CK-MB in diagnosing myocardial injury were 90%,66.7% and 83.3%,respectively. The sensitivity of GPBB was significantly higher than that of cTnI(χ~2 = 4.812,P < 0.05),with no statistical difference between GPBB and CK-MB(χ~2 = 0.577,P > 0.05). The specificities of GPBB,cTnI and CK-MB in diagnosing myocardial injury were 88.2%,91.2% and 67.6%,respectively. The specificity of GPBB was significantly higher than that of CK-MB(χ~2 = 4.191,P < 0.05),with no statistical difference between GPBB and cTnI(χ~2 = 0.159,P > 0.05). Plasma GPBB levels were significantly higher in neonates with severe asphyxia(14.67,6.09 ng/ml)than those with mild asphyxia(5.61,3.56 ng/ml)and control group(P < 0.01). GPBB levels were higher in mildly asphyxiated neonates than those in control neonates,but with no statistical significance(P > 0.05). Spearman rank correlation analysis showed that plasma GPBB levels positively correlated with the cloudiness of the amniotic fluid(r = 0.500,P < 0.001)and negatively correlated with Apgar score(r = -0.520,P < 0.001). Conclusions This study indicated that GPBB can be used as an ideal biomarker of myocardial ischemia injury in neonates with asphyxia. The determination of GPBB in early neonatal period is useful in detecting and assessing the severity of myocardial injury.

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