Intelligent analysis of excitation-emission matrix fluorescence fingerprint to identify and quantify adulteration in camellia oil based on machine learning.

Camellia oil (CAO) is a premium edible vegetable oil with medical value and biological activity, but it is susceptible to adulteration. Therefore, the demand for intelligent analysis to decipher the category and proportion of adulterated oil in CAO was the main driver of this work. Excitation-emission matrix fluorescence (EEMF) spectra of 933 vegetable oil samples were characterized by a chemometric method to obtain chemically meaningful information. Authenticity identification models were constructed using four machine learning methods to realize the discrimination of oil species adulterated in CAO mixtures. Meanwhile, quantitative models were established aiming at the fraud of CAO proportion in blended oil. Results showed that the specially constructed CNN obtained the optimal performance when evaluating unseen real-world samples, with a classification accuracy of 95.8% and 92.2%, and mean-absolute quantitative errors between 2.6 and 6.7%. Therefore, EEMF fingerprints coupled with machine learning are expected to provide intelligent and accurate analysis for authenticity detection of CAO.

Chemometrics-enhanced high-performance liquid chromatography-diode array detection strategy to quantify protoberberine alkaloids in varying Coptidis Rhizoma-related medicines.

Quantitation of protoberberine alkaloids is an essential guarantee for efficacy control and medication safety of Coptidis Rhizoma (CR) related medicines. Traditional univariate chromatography faced challenges with co-elution, unknown interferences, and retention time shift when analyzing isomeric analytes in varying sample matrices. We presented a chemometrics-enhanced high-performance liquid chromatography-diode array detection (HPLC-DAD) strategy for simultaneous quantification of six protoberberine alkaloids and processed multi-channels chromatographic-spectral data with four second-order calibration algorithms. Chromatographic conditions were firstly optimized. Four groups of predicted samples were modeled individually with the designed calibration set. Mathematical resolutions were then obtained, and pseudo-univariate regression gave the quantitative concentration of each analyte. Four models were scored on fit, linearity, recovery, and robustness, where alternating trilinear decomposition assisted multivariate curve resolution (ATLD-MCR) exhibited an optimal and stable performance. Besides, the resolved spectra presented high consistency with the actual spectra (r≥0.9993). Limits of quantification (LOQ) fully met the pharmacopoeia stipulation and were 0.17, 0.60, 0.19, 0.74, 0.15, and 0.38 µg mL-1 for columbamine, epiberberine, jatrorrhizine, coptisine, palmatine, and berberine, respectively. The importance of this strategy is to exploit collinearity resolution and additional selectivity that permit accurate quantitation at poor chromatographic resolutions, avoiding individual pretreatment and HPLC optimizations for different samples. This study provides a universal alternative for routine quality assessment of protoberberine alkaloids in CR-related medicines.

[Swimming plus medication reduces the expressions of cytokines in rats with chronic abacterial prostatitis].

OBJECTIVE: To observe the effects of swimming plus medication on the expressions of cytokines in rats with chronic abacterial prostatitis (CAP). METHODS: Forty healthy adult male SD rats were randomly divided into five groups of equal number, normal control, CAP model control, medication, exercise therapy, and exercise + medication. The CAP model was made by Xiaozhiling injection, and at 7 days after modeling, the rats in the medication and exercise + medication groups were treated intragastrically with Qianlie Shutong Capsules (0.016 g/ml) at 20 ml per kg of the body weight qd, those in the exercise therapy and exercise + medication groups were made swim at a regular time once a day, 35 minutes on the first day and 5 minutes more on the second until 50 minutes once, for 4 successive weeks, and those in the normal control, model control and exercise therapy groups received normal saline only. After 14 and 28 days of treatment, all the rats were killed and their prostates harvested for observation of histopathological changes and determination of the expressions of TNF- α, IL-1ß and IL-6 in the prostatic tissue homogenate by ELISA. RESULTS: After 14 days of treatment, the expression levels of TNF-α, IL-1ß and IL-6 were significantly elevated in the groups of CAP model control (ï¼»183.08±8.07ï¼½ pg/ml, ï¼»57.55±3.53ï¼½ pg/ml and ï¼»256.15±13.95ï¼½ ng/L), medication (ï¼»118.49±8.06ï¼½ pg/ml, ï¼»42.64±4.64 ï¼½ pg/ml and ï¼»200.74±9.33ï¼½ ng/L), exercise therapy (ï¼»169.63±10.64ï¼½ pg/ml, ï¼»50.45±5.71ï¼½ pg/ml and ï¼»245.23±6.49ï¼½ ng/L), and exercise + medication (ï¼»107.82±7.81ï¼½ pg/ml, ï¼»40.35±6.93ï¼½ pg/ml and ï¼»187.04±10.85ï¼½ ng/L) as compared with those in the normal control (ï¼»20.36±1.82ï¼½ pg/ml, ï¼»14.64±1.91ï¼½ pg/ml and ï¼»70.58±2.09ï¼½ ng/L) (P<0.05). At 28 days, the levels of TNF- α, IL-1ß, IL-6 were remarkably lower in the exercise + medication group (ï¼»29.30±3.78ï¼½ pg/ml, ï¼»16.91±1.24ï¼½ pg/ml and ï¼» 88.65±6.74ï¼½ ng/L) than in the medication group (ï¼»39.67±3.19ï¼½ pg/ml, ï¼»26.27±3.49ï¼½ pg/ml and ï¼»110.26±6.33ï¼½ ng/L) (P<0.05) and close to those of the normal control group (ï¼»19.34±1.76ï¼½ pg/ml, ï¼»13.68±1.06ï¼½ pg/ml and ï¼»71.34±2.50ï¼½ ng/L). During the treatment, no obvious pathological changes were found in the prostate tissue of the normal control rats, while significant chronic prostatic inflammation was observed in the CAP models, and the inflammation was relieved in different degrees after intervention, most significantly in the exercise + medication group. CONCLUSIONS: Swimming can relieve prostatic inflammation and swimming plus medication can effectively reduce the expressions of cytokines and alleviate histological damage in the prostatic tissue of CAP rats.

Regulation of demethylation and re-expression of RASSF1A gene in hepatocellular carcinoma cell lines treated with NCTD in vitro.

BACKGROUND: Hepatocellular carcinoma, a lethal malignant neoplasm with poor prognosis, has dismal results of surgical resection and chemoradiotherapy. Norcantharidin (NCTD), the demethylated analog of cantharidin derived from a traditional Chinese medicine, Mylabris, has been used in the treatment of cancer. However, the detailed mechanisms underlying this process are generally unclear. PURPOSE: The aim of this study was to investigate the mechanism of NCTD-induced apoptosis in HepG2 cells. MATERIALS AND METHODS: Human HepG2 cell lines were treated with NCTD at different concentrations (2.50, 5.00, 10.00, 20.00, 40.00 µg/mL) for 24 hours. Cell proliferation was evaluated by measurement of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The methylation levels of RASSF1A (Ras-association domain family 1 A) in HepG2 cells were detected by methylation-specific PCR (MSP). The mRNA levels of RASSF1A in HepG2 cells were detected by real-time fluorescent quantitative PCR (RT-PCR). The levels of RASSF1A protein expression of HepG2 cells were detected by Western blotting assay. RESULTS: The inhibition of cell proliferation was observed when treated with NCTD at concentrations (2.5 µg/mL), and as concentration increased, the proliferation of HepG2 cells was markedly inhibited by NCTD in dose-dependent manners. The levels of methylation of RASSF1A decreased at the increasing concentration of 10, 20 and 40 µg/mL. The levels of RASSF1A mRNA and protein were decreased when treated with NCTD at the concentrations of 10, 20 and 40 µg/mL, which were also in a dose-dependent manner. CONCLUSION: NCTD can reverse the methylation state of RASSF1A gene and induce its re-expression, which will provide the theoretical basis for the clinical practice.