The molecular mechanism of action for the potent antitumor component extracted using supercritical fluid extraction from Croton crassifolius root.

ETHNOPHARMACOLOGICAL RELEVANCE: The root of Croton crassifolius has been used as a traditional Chinese medicine (TCM), called Radix Croton Crassifolius, and commonly known as "Ji Gu Xiang" in Chinese. Its medicinal value has been recorded in several medical books or handbooks, such as "Sheng Cao Yao Xing Bei Yao", "Ben Cao Qiu Yuan" and "Zhong Hua Ben Cao". It has been traditional employed for treating sore throat, stomach-ache, rheumatism and cancer. AIM OF THE STUDY: At present, there are limited studies on the evaluation of low-polarity extracts of roots in C. crassifolius. Consequently, the aim of this study was to evaluate the antitumor effect of the low-polarity extract of C. crassifolius root. MATERIALS AND METHODS: Extracts were obtained by supercritical fluid extraction. The extracts were tested for antitumor effects in vitro on several cancer cell lines. A CCK-8 kit was used for further analysis of cell viability. A flow cytometer and propidium iodide staining were used to evaluate the cell cycle and apoptosis. Hoechst staining, JC-1 staining and the fluorescence probe DCFH-DA were used to evaluate apoptotic cells. Molecular mechanisms of action were analyzed by quantitative RT‒PCR and Western blotting. Immunohistochemistry was used for the evaluation of xenograft tumors in male BALB/c mice. Finally, molecular docking was employed to predict the bond between the desired bioactive compound and molecular targets. RESULTS: Eleven diterpenoids were isolated from low-polarity C. crassifolius root extracts. Among the compounds, chettaphanin II showed the strongest activity (IC50 = 8.58 µM) against A549 cells. Evaluation of cell viability and the cell cycle showed that Chettaphanin II reduced A549 cell proliferation and induced G2/M-phase arrest. Chttaphanin II significantly induced apoptosis in A549 cells, which was related to the level of apoptosis-related proteins. The growth of tumor tissue was significantly inhibited by chettaphanin II in experiments performed on naked mice. The antitumor mechanism of chettaphanin II is that it can obstruct the mTOR/PI3K/Akt signaling pathway in A549 cells. Molecular docking established that chettaphanin II could bind to the active sites of Bcl-2 and Bax. CONCLUSIONS: Taken together, the natural diterpenoid chettaphanin II was identified as the major antitumor active component, and its potential for developing anticancer therapies was demonstrated for the first time by antiproliferation evaluation in vitro and in vivo.

Left ventricular entropy is a novel predictor of major adverse cardiac events (MACE) in patients with coronary atherosclerosis: a multi-center study.

OBJECTIVES: To investigate the incremental prognostic value of left ventricular (LV) entropy in a large multi-center population with coronary atherosclerotic heart disease (CAD). BACKGROUND: Current risk stratification of patients with CAD is imprecise and not accurate enough. METHODS: A total of 314 CAD patients who underwent cardiovascular magnetic resonance (CMR) late gadolinium enhancement (LGE) at two medical centers in China between October 2015 and July 2022 were included in this study. Additionally, the 193 patients under 3.0-T field also underwent CMR T1 mapping. LV entropy and extracellular volume (ECV) were calculated from the LGE image of LV myocardium, and major adverse cardiac events (MACEs) were analyzed. RESULTS: Among 314 patients, 110 experienced MACE during a median follow-up of 13 months. The risk of MACE was significantly increased in the high entropy group (log-rank p < 0.001). Entropy maintained an independent association with MACE in a multivariable model including left ventricular ejection fraction (LVEF) and LGE (HR = 1.78; p = 0.001). In addition, the primary endpoint events prognostic value was significantly improved by adding LV entropy to the baseline multivariable model (C-statistic improvement: 0.785-0.818, Delong test: p = 0.001). Similarly, among 193 3.0-T field patients, adding LV entropy to the multivariable baseline model significantly improved the prognostic value of the model for MACE (C-statistic improvement: 0.820-0.898, Delong test: p = 0.004). CONCLUSION: CMR-assessed LV entropy is a powerful independent predictor of MACE in patients with CAD, incremental to common clinical and CMR risk factors, including LVEF, LGE, Native T1, and ECV. CLINICAL RELEVANCE STATEMENT: Left ventricular entropy is a powerful independent predictor of major adverse cardiac events in patients with coronary atherosclerotic heart disease, incremental to common clinical and cardiac magnetic resonance risk factors. KEY POINTS: • Left ventricular entropy, a novel cardiac magnetic resonance parameter of myocardial heterogeneity, demonstrated a robust prognostic association with major adverse cardiac events beyond guideline-based, clinical risk markers. • Entropy can have an important role in the primary prevention of major adverse cardiac events in patients with coronary atherosclerotic heart disease. • Compared with late gadolinium enhancement, extracellular volume, and native T1, entropy could be used to more comprehensively characterize the heterogeneity of left ventricular myocardium.

Brain structural changes in preschool children with MRI-negative epilepsy.

PURPOSE: To investigate abnormalities in cortical and subcortical structures of the brain in preschool children with MRI-negative epilepsy. METHODS: Cortical thickness, cortical mean curvature, cortical surface area, cortical volume, and volumes of subcortical structures were measured using Freesurfer software in preschool children with epilepsy and age-matched controls. RESULTS: Findings showed cortical thickening in the left fusiform gyrus, left middle temporal gyrus, right suborbital sulcus, and right gyrus rectus, and cortical thinning mainly in the parietal lobe of preschool children with epilepsy compared to controls. The difference in cortical thickness in the left superior parietal lobule remained after correction for multiple comparisons and was negatively correlated with duration of epilepsy. Cortical mean curvature, surface area, and volume were mainly altered in the frontal and temporal lobes. Changes in mean curvature in the right pericallosal sulcus were positively correlated with age at seizure onset, and changes in mean curvature in the left intraparietal sulcus and transverse parietal sulcus were positively correlated with frequency of seizures. There were no significant differences in the volumes of the subcortical structures. CONCLUSION: Changes in preschool children with epilepsy occur in the cortical rather than subcortical structures of the brain. These findings further our understanding of the effects of epilepsy in preschool children and will inform management of epilepsy in this patient population.

Early detection of myocardial fibrosis in cardiomyopathy in the absence of late enhancement: role of T1 mapping and extracellular volume analysis.

OBJECTIVES: To analyze myocardial fibrosis in dilated cardiomyopathy (DCM) patients with no late gadolinium enhancement (LGE) on cardiac magnetic resonance (CMR) using T1 mapping and extracellular volume (ECV) and investigate the potential correlation with left ventricular (LV) dilation and dysfunction. METHODS: The study included 41 DCM patients without LGE and 79 healthy controls. T1 and ECV were compared between the two groups using multivariable logistic regression analysis. The correlations between histological and functional parameters were evaluated using Pearson's correlation. RESULTS: Mean native myocardial T1 and ECV were significantly higher in the DCM group compared to controls (p ≤ 0.001, respectively). Multivariable logistic regression revealed that ECV (mean, minimum), LV ejection fraction (LVEF), and LV end-diastolic diameter (LVEDD) were independent discriminators for LGE-negative DCM; the area under the curve (AUC) of LVEF, LVEDD, ECV mean, and ECV minimum were 0.97, 0.96, 0.88, and 0.68, respectively. In the DCM group, LVEDD and LVEF were positively and negatively correlated with ECV, respectively. LVEDV index and LV end-systolic volume (LVESV) index were positively correlated with native-T1 and ECV, and the absolute value of LV global strain had a negative correlation with ECV. CONCLUSIONS: Early myocardial fibrosis in DCM could be detected by prolonged native T1 and elevated ECV despite the absence of LGE on CMR. Moreover, the change of histological characteristics of myocardium in DCM was correlated with LV dilation and dysfunction. KEY POINTS: • At an early stage, patients with DCM may have myocardial fibrosis despite the absence of LGE. • T1 mapping and ECV are efficient methods for early detection of potential myocardial fibrosis. • Increased native T1 and ECV are correlated with left ventricular dilation and dysfunction in LGE-negative DCM patients.

Remodeling of occluded internal carotid artery in vessel wall magnetic resonance imaging.

OBJECTIVE: The purpose of the present study was to investigate the remodeling pattern of the extracranial occluded internal carotid artery (OICA) by vessel wall magnetic resonance imaging (VWI). METHODS: Thirty-nine atherosclerotic OICAs from 32 consecutive cases underwent 3-Tesla VWI to acquire pre- and post-contrast T1-weighted two-dimensional fluid-attenuated inversion recovery fast spin echo sequences. 25 symptomatic CAs exhibited ipsilateral downstream cerebral ischemia or ophthalmic artery embolism within last three months. The 14 remaining CAs were asymptomatic. Twenty-four CAs from 22 patients with atherosclerosis but no stenosis were recruited as control group. The outer wall area (OWA) was calculated based on the outer contour of the carotid artery drawn on the pre-contrast VWI. Negative remodeling was defined as a lower OWA compared to that of control group. RESULTS: Clinical characteristics including age, sex and vascular risk factors showed no significant difference between the occluded and control group. However, the OWA was lower in the occluded group than in the control group (0.63 versus 0.90 cm2, p = 0.004). For all OICAs, the OWA was larger in symptomatic cases than asymptomatic cases (0.71 versus 0.49cm2, p = 0.025). Using a cutoff value of 0.44, the sensitivity and specificity of OWA for detecting symptomatic OICA were 0.88 and 0.57, respectively. Heterogeneous signal intensity and enhancement were more often observed at the proximal than the distal segment of occlusion (p < 0.001). The inter-observer agreement regarding the evaluation of VWI characteristics was desirable (κ = 0.805 ∼ 0.847). CONCLUSIONS: Negative remodeling is prevalent in OICA, especially in asymptomatic cases.

Diffusion-weighted imaging for the left hepatic lobe has higher diagnostic accuracy for malignant focal liver lesions.

BACKGROUND: This study was conducted to investigate whether apparent diffusion coefficient (ADC) measurements by dividing the liver into left and right hepatic lobes may be utilized to improve the accuracy of differential diagnosis of benign and malignant focal liver lesions. MATERIALS AND METHODS: A total of 269 consecutive patients with 429 focal liver lesions were examined by 3-T magnetic resonance imaging that included diffusion-weighted imaging. For 58 patients with focal liver lesions of the same etiology in left and right hepatic lobes, ADCs of normal liver parenchyma and focal liver lesions were calculated and compared using the paired t-test. For all 269 patients, ADC cutoffs for focal liver lesions and diagnostic accuracy in the left hepatic lobe, right hepatic lobe and whole liver were evaluated by receiver operating characteristic curve analysis. RESULTS: For the group of 58 patients, mean ADCs of normal liver parenchyma and focal liver lesions in the left hepatic lobe were significantly higher than those in the right hepatic lobe. For differentiating malignant lesions from benign lesions in all patients, the sensitivity and specificity were 92.6% and 92.0% in the left hepatic lobe, 94.4% and 94.4% in the right hepatic lobe, and 90.4% and 94.7% in the whole liver, respectively. The area under the curve of the right hepatic lobe, but not the left hepatic lobe, was higher than that of the whole liver. CONCLUSIONS: ADCs of normal liver parenchyma and focal liver lesions in the left hepatic lobe were significantly higher than those in the right hepatic lobe. Optimal ADC cutoff for focal liver lesions in the right hepatic lobe, but not in the left hepatic lobe, had higher diagnostic accuracy compared with that in the whole liver.