Regression Area Research Articles

An integrated strategy for deciphering quality markers of Terminaliae Belliricae Fructus based on a three-dimensional characteristic model

Terminalia bellirica (Gaertn.) Roxb. is an ethnomedicinal plant that has been utilized in Tibetan and traditional Chinese medicine (TCM). Nevertheless, its quality standard officially listed in the Chinese Pharmacopoeia does not include any content determination of the indicator components of Terminaliae Belliricae Fructus, which constrains the effective quality evaluation of medicinal material and related products. In this paper, a three-dimensional “content-pharmacokinetics-pharmacology” network strategy was developed to identify the quality markers (Q-markers) of Terminaliae Belliricae Fructus. in terms of “measurability”, “traceability” and “effectiveness”. Chromatographic fingerprint analysis was performed to outline its chemical contour, and identify the differential components of 17 batches of Terminaliae Belliricae Fructus. combined with multivariate statistics analysis and UPLC-QTOF-MS analysis. Serum pharmacochemistry analysis was implemented on rats, and 25 prototype components absorbed into the blood were identified. By network pharmacology analysis, a component-disease-target-pathway network was constructed, thus validating the effectiveness of the chemical components of Terminaliae Belliricae Fructus. Afterwards, the above screened candidate components were put into construction of three-dimensional "radar chart". According to the calculated regression area (RA) and coefficient of variation (CV) values, the potential Q-markers was determined, followed by “specificity” evaluation. Ultimately, ellagic acid (EA), chebulagic acid (CHA), gallic acid (GA), chebulinic acid (CA), corilagin (CO) and chebulanin (CH) were specified as the Q-markers of Terminaliae Belliricae Fructus. Owing to high content, good pharmacokinetic property, high pharmacological activities and specificity. The screened Q-markers could offer a scientific foundation for the quality control of Terminaliae Belliricae Fructus, and the proposed strategy is demonstrated to be reliable and feasible for deciphering Q-markers of TCM.

A mathematical multiple-dimensional strategy for Q-markers identification based on “five principles”: Tianshu Capsule for migraine treatment as an example

Ethnopharmacological relevanceQuality control is a critical element for Traditional Chinese medicine (TCM). Due to the varied chemical components, mechanisms of action, and pharmacological functions in TCM, ensuring quality is more challenging compared to chemical drugs. Then, the concept of quality markers (Q-markers) was proposed and ideal Q-markers for TCM prescriptions need to compliant with “five principles”, including pharmacological effectiveness, specificity, transfer and traceability, measurability, and prescription compatibility. Aim of the studyTo establish a mathematical multiple-dimensional “spider-web” strategy and identify the Q-markers of Tianshu capsule (TSC), a Chinese patent medicine for the treatment of migraine, following the “five principles” rules. Materials and methodsQ-marker candidates of TSC were firstly screened according to the HPLC fingerprints. Their contents in 10 batches of TSC and stabilities under high temperature, high humidity and in work solutions were determined quantitatively by HPLC-UV (measurability). Their existences in Gastrodiae Rhizoma, Chuanxiong Rhizoma, TSC, rat plasma and brain samples were investigated using HPLC-Q-TOF/MS (transfer and traceability). Their anti-migraine efficacies were evaluated by network pharmacology and mice hot-plate analgesia test; and their relationships with the property (flavor) of Gastrodiae Rhizoma or Chuanxiong Rhizoma were studied by molecular docking (effectiveness). Their contributions were defined based on their herb source according to the compatibility theories of Da Chuan Xiong Fang (compatibility). Their biosynthetic pathways were studied, and their frequencies detected in different plant families were calculated (specificity). Finally, an eight dimensional “spider-web” mode was developed for 10 components, and the regression area (RA) and the coefficient variation (CV) of each candidate were calculated after data normalization. ResultsTen components including gastrodin, parishin E, chlorogenic acid, ferulic acid, isochlorogenic acid A, senkyunolide I, H, A, Z-ligustilide and levistilide A were selected and evaluated as the Q-marker candidates. The results showed that gastrodin, senkyunolide I, and senkyunolide A had the higher RA and lower CV than other compounds with the established “spider-web” mode, indicating that they could be used as the Q-markers of TSC. ConclusionThe multi-dimensional “spider-web” mode based on “five principles” was firstly applied to identify the Q-markers of TSC, and it can be used as a practical strategy to discover Q-markers of other compounded prescriptions.

The discovery of Q-markers of Qiliqiangxin Capsule, a traditional Chinese medicine prescription in the treatment of chronic heart failure, based on a novel strategy of multi-dimensional “radar chart” mode evaluation

BackgroundQiliqiangxin Capsule (QLQX), a traditional Chinese medicine (TCM) prescription, is especially used for clinical treatment of chronic heart failure (CHF) in China. However, the holistic quality control of QLQX has not been well established due to lack of system research on the quality marker (Q-marker). PurposeIn this study, a new strategy of multi-dimensional “radar chart” mode was proposed to overcome the problem that traditional methods cannot evaluate the multiple properties of Q-markers comprehensively and visually, and the strategy was successfully applied to discover the Q-markers of QLQX. MethodsFirst, nineteen prototypes that entered the in vivo systemic circulation were selected out as the candidate Q-markers based on our previous studies of chemical and in vivo metabolic profiles. Then, their contents in QLQX were quantitatively analyzed by UHPLC-MS/MS, and the bioactivities on the H9c2 cardiomyocytes cell model was evaluated. The network of in vivo component-target closely related to CHF was further constructed. Finally, a multi-dimensional “radar chart” mode was developed and corresponding Regression Area (RA) and Coefficient Variation (CV) were calculated after data standardization and integration visually based on the Q-marker related multiple characteristics (including the compatibility contribution of herbal medicines, the content, the bioactivity, the in vivo predicted bioavailability and the degree of network pharmacology of candidate components in the TCM prescription). ResultsBy comparison of RA and CV of the chemicals in the “radar chart”, seven compounds mainly from King and Minister herbs (songorin, calycosin-7-O-β-D-glucopyranoside, astragaloside, tanshinone IIA, ginsenoside Re, hesperidin and alisol A) were screened out as the Q-markers of QLQX, showing the reasonable compatibility contribution and high content in QLQX, preferable pharmacological effect on CHF, as well as good bioavailable characteristics and high target hits in system pharmacology. ConclusionThe Q-marker discovery of QLQX in this study laid an important foundation for its quality control improvement, and the mode standardized the abstract definitions of Q-marker and realized the comprehensive assessment of multiple properties of Q-marker in TCM prescriptions, which has a reference value for revealing the Q-marker in the quality control researches of TCM prescriptions.

The biology and prognostic value of lymphatic vessel density (LD) and lymphatic invasion (LI) in regression in melanoma

9017 Background: Regression in melanoma is characterized by increased vascularity, lymphocytic infiltrate and fibroplasia in the papillary dermis, accompanied by the absence (complete regression, CoR) or presence (partial regression, PaR) of melanoma cells in the epidermis. The prognostic value of regression is controversial. We noticed that LD and LI were increased in the areas of regression (AR) or areas with brisk lymphocytic infiltration (AB). Our goal was to clarify the prognostic value of regression in melanoma. Methods: Dual immunohistochemical staining was done using antibodies to podoplanin (lymphatic vessels) and S100 (melanoma cells) on paraffin tissues from 321 patients with vertical growth phase (VGP) primary melanomas who had 10 years or more of follow-up. LD in AR (both CoR and PaR) was compared with that of normal dermis adjacent and distant, as well as LD in the AB. LI in these areas was also scored. Unadjusted and adjusted hazard rates were obtained from univariate and multivariate Cox models for time to melanoma-specific death using established melanoma prognostic factors. Results: 116 patients (36%) had regression: 75 CoR (23%) and 41 PaR (13%). LD significantly decreased stepwise from CoR (mean ± se, 23.7 ± 2.7) to PaR (15.5 ± 1.1), adjacent normal dermis (7.3 ± 0.28) and distant normal dermis (5.4±0.31) and it was significantly elevated in the AB (18.5±0.78). Melanomas with CoR had the highest percentage of LI in both AR and AB. In addition, the percentage of LI in AB was highest for men and for those with VGP tumor infiltrating lymphocytes (TILs). Both high LD in AR and more LI in AB were associated with poor prognosis (p=0.004 and p=0.002, respectively). Six factors were significant in the final multivariate model: LI in AB (HR=2.3), LD in AR (HR=1.04), thickness (HR=1.44), axial (HR=7.7), ulceration (HR=2.5) and no VGP TILs (HR=2.8). Conclusions: AR and AB were associated with increased LD and higher incidence of LI in primary melanomas. LD and LI in AR or AB are independent prognostic factors. Our data suggest that the effects of regression on prognosis are mediated at least in part through lymphangiogenesis and LI. No significant financial relationships to disclose.