OBJECTIVE: Coix lacryma-jobi, a highly regarded Asian herb widely used in traditional Chinese medicine, is recognized for its dual benefits in promoting overall health and treating various diseases. While it exhibits moderate anticancer efficacy when used alone, this study investigated the enhanced anticancer potential of raw and cooked Coix lacryma-jobi var. lacryma-jobi (CL) seed extracts in combination with sorafenib against HCT116 and HepG2 cancer cell lines. The combination of sorafenib with other anticancer agents, including natural extracts, has garnered significant attention as a promising strategy for developing more effective cancer therapies. METHODS: Dry powders of raw (R) and cooked (C) CL seeds, obtained from a local commercial source in Thailand, were extracted and fractionated using ethanol (E), dichloromethane (D), ethyl acetate (A), and water (W) to produce eight fractions: CLRE, CLCE, CLRD, CLCD, CLRA, CLCA, CLRW, and CLCW. The coixol content in raw and cooked seed extracts was quantified and expressed as μg of coixol per gram of extract. The cytotoxic effects of these fractions were evaluated against HCT116 and HepG2 cells using the MTT assay. Fractions demonstrating the most significant cytotoxic responses were combined with sorafenib to evaluate their synergistic effects. Apoptosis induction and mitochondrial membrane potential (MMP) were assessed, and the underlying mechanism of apoptosis was explored by analyzing reactive oxygen species (ROS) generation and antioxidant protein expression levels. Additionally, the combination treatment's effect on the phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT)/mechanistic target of rapamycin (mTOR) pathway was investigated. RESULTS: One gram of CLCE and CLCD extracts contained higher coixol levels (7.02 μg and 9.69 μg, respectively) compared to CLRE and CLRD (2.66 μg and 5.96 μg, respectively). Coixol content in CLRA, CLRW, and CLCW fractions was undetectable under the study conditions. All extract fractions exhibited IC₅₀ values exceeding 1 mg/mL after 24- and 48-hour incubations with HCT116 and HepG2 cells, indicating limited cytotoxicity when used independently. CLRD and CLCD fractions were selected for combination studies at a concentration of 1 mg/mL, combined with sub-IC₅₀ concentrations of sorafenib to minimize its side effects. This combination significantly increased cytotoxicity, inducing apoptosis in HCT116 and HepG2 cells by elevating ROS levels and reducing the expression of superoxide dismutase 2 and catalase. Furthermore, the combination treatment downregulated the PI3K/AKT/mTOR pathway, indicating a targeted anticancer mechanism. CONCLUSION The combination of CLCD with sorafenib demonstrates significant potential as a strategy for future anticancer therapies. This CL seed extract, cultivated and commercially available in Thailand, shows promise as a natural supplement to enhance the efficacy of chemotherapy in upcoming clinical anticancer applications.

Objective To evaluate the relationship between the expression of microRNA-146a (miR-146a) in liver tissue and the inflammatory hepatic injury induced by ischemia/reperfusion (I/R) in rats. Methods One hundred and forty-four Sprague-Dawley (SD) rats were randomly divided into three groups: control (group N), sham operation (group S) and group I/R. Each group was subdivided into four subgroups (n = 12), and different substances were respectively injected intravenously to rats in different subgroups at 1 hour before the experiment: 220 μL physiological saline (group A), 20 μL miR-146a mimic + 200 μL physiological saline (group B), 20 μL miR-146a mimic + 200 μL ultrasound microbubble contrast agent (group C) and 20 μL miR-146a inhibitor + 200 μL ultrasound microbubble contrast agent (group D). Before the experiment and after experiment for 24 hours, the plasma concentrations of alanine aminotransferase (ALT), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected, the reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of miR-146a in liver tissue, and Western Blot was applied to detect protein expressions of Toll-like receptor 4 (TLR4), IL-1 receptor associated kinase 1 (IRAK-1), IL-6 and TNF-α, and the pathological hepatic cell injury was observed. Results Before the experiment and 24 hours after experiment in various subgroups of N and S groups, there were no statistical significant differences in the plasma concentrations of ALT, IL-6 and TNF-α, and the expression of miR-146a level and the protein expressions of TLR4, IRAK-1, IL-6 and TNF-α in liver tissues; the pathological examination also did not show any obvious hepatic cell injury. After the experiment for 24 hours: compared to the group S, the liver tissue miR-146a expression was significantly decreased in the subgroups A and D of group I/R (miR-146a/U6nsRNA: 0.51±0.13, 0.22±0.09 vs. 1.01±0.02, both P < 0.01), and the plasma concentrations of ALT, IL-6 and TNF-α and the protein expressions of TLR4, IRAK-1, IL-6 and TNF-α in liver tissues were significantly increased [ALT (U/L): 103.23±26.64 vs. 44.16±18.55, 176.46±7.26 vs. 49.74±6.83, IL-6 (μg/L): 64.28±16.19 vs. 17.68±7.54, 88.49±3.23 vs. 15.58±2.38; TNF-α (μg/L): 31.28±2.57 vs. 5.58±3.35, 59.12±8.74 vs. 5.27±1.37; TLR4/GAPDH: 2.43±0.36, 3.23±0.71 vs. 0.96±0.24, IRAK-1/GAPDH: 2.34±0.52, 3.14±0.63 vs. 0.76±0.21, IL-6/GAPDH: 1.01±0.22, 1.11±0.16 vs. 0.98±0.37, TNF-α/GAPDH: 2.05±0.48, 2.86±0.27 vs. 0.59±0.16, all P < 0.01], moreover, the hepatic pathological lesions were obvious; the liver tissue expression of miR-146a was significantly increased after being transfected with miR-146a mimic in subgroups B and C of group I/R (miR-146a/U6nsRNA: 1.56±0.31, 2.40±0.53 vs. 1.01±0.02, both P < 0.01), especially in group C combined with ultrasound microbubble (P < 0.01). However, the protein expressions of TLR4, IRAK-1, IL-6 and TNF-α in liver tissues were significantly decreased (TLR4/GAPDH:0.77±0.18, 0.65±0.27 vs. 0.96±0.24, IRAK-1/GAPDH: 0.61±0.14, 0.47±0.20 vs. 0.76±0.21, IL-6/GAPDH:0.80±0.13, 0.54±0.22 vs. 0.98±0.37, TNF-α/GAPDH: 0.41±0.14, 0.16±0.03 vs. 0.59±0.16; all P < 0.01), and the expressions were more significant in the group C combined with ultrasound microbubbles (P < 0.01), and the hepatic pathological damage was mild, however, the plasma concentrations of ALT, IL-6 and TNF-α were of no statistical significant differences. Conclusion Ultrasound microbubble can efficiently transfect miR-146a mimic and inhibitor into the liver tissue, and miR-146a may negatively regulate the I/R inflammatory liver injury mediated by TLR signaling pathway.

OBJECTIVE:To establish a method for the determining contents of 2 lignan components[dehydrodiisoeugenol and 2,3-dihydro-7-methoxy-2-(3,4-methylened ioxyphenyl)-3-methyl-5-(E)-propenyl-benzofuran(referred to“lignanoid 2”)]. METH-ODS:HPLC method was adopted. The column was Elite C18 with the mobile phase of water-methanol(gradient elution)at the flow rate of 1.0 ml/min;the detection wavelength was 225 nm and the column temperature was 30 ℃. The sample size was 20 μl. RE-SULTS:There was a good linear relationship between sample quantity and the peak area in the range of 0.202-2.02 μg(r=0.999 9) and 0.204-2.04 μg(r=0.999 9)for 2 lignan components. The RSD of precision,stability and repeatability tests were less than 2%with the average recovery of 101.54%(RSD=0.60%,n=6)and 99.43%(RSD=1.09%,n=6). CONCLUSIONS:The method is simple,sensitive and accurate,and can be used for the quantization determination of dehydrodiisoeugenol and lignanoid 2 in nut-meg-5.

AIM: To establish a convenient liquid chromatography-internal standard method for determining ligustilide and 6-gingerol in Kuanxuean Soft Capsule (Radix Angelicae Sinensis, Rhizoma Zingiberis Preparatum, etc.) METHODS: Ligustilide, 6-gingerol and the internal standard naphthalene were separated on a Kromasil C_ 18 column. The mobile phase consisted of methanol-trifluroacetic acid (67 ∶ 33, V/V) and was determined at 218 nm. The content of ligustilide and 6-gingerol was calculated by the internal standard method. RESULTS: The recovery of ligustilide was 98.64% (RSD=1.58%); the recovery of 6-gingerol was 97.25% (RSD=1.40%). Result of the standard curve method accords with that of internal standard method. CONCLUSION: The method is sensitive, stable, practical, and suitable for the quality control of Kuanxuean Soft Capsule.

Objective: To study the application of macroporous resin combined with membrane in the refinement of traditional Chinese medicine. Methods: The extracts of single herbal drug and prescriptions were absorpted with macroporous resin, following by microfiltrated respectively, and then detected the extracts and analysized the quanity of effective components. Results: Through macroporous resin combined with membrane, the quantity of effective components could be improved significantly, and the method was very effective for refinement of traditional Chinese medicine. Conclusion: Application of macroporous resin combined with membrane has great prospect in the refinement of traditional Chinese medicine.

In order to discover some sensitive parameters of eight diastolic function indexes and establish the discriminant equation, twenty-three patients with coronary heart disease (CAD) and eighteen normal adults were studied by radionuclide, Doppler and apexcardiography. The results showed that eight indexes were signicantly different between CAD and normal groups. The discriminant equation was: Z=0.21X1+23.86X2-22.88X3-0.18X4+2.83X5+ 2.06X6+66.86X7+1.66X8. The discriminant score was 42.99. The discriminant function was 100%. The contributive rates of eight indexes were Ev/Av, 1/3FF, A/E-O, PFR, DATI, EDC, IRT ;and l/3FFd from large to small value, respectively. It is concluded that Doppler Ev/Av is the most sensitive diastolic function index of CAD.