Effect of Chestnut (Castanea Mollissima Blume) Bur Polyphenol Extract on Shigella dysenteriae: Antibacterial Activity and the Mechanism.

Shigella dysenteriae is a highly pathogenic microorganism that can cause human bacillary dysentery by contaminating food and drinking water. This study investigated the antibacterial activity of chestnut bur polyphenol extract (CBPE) on S. dysenteriae and the underlying mechanism. The results showed that the minimum inhibitory concentration (MIC) of CBPE for S. dysenteriae was 0.4 mg/mL, and the minimum bactericidal concentration (MBC) was 1.6 mg/mL. CBPE treatment irreversibly disrupted cell morphology, decreased cell activity, and increased cell membrane permeability, cell membrane depolarization, and cell content leakage of S. dysenteriae, indicating that CBPE has obvious destructive effects on the cell membrane and cell wall of S. dysenteriae. Combined transcriptomic and metabolomics analysis revealed that CBPE inhibits S. dysenteriae by interfering with ABC protein transport, sulfur metabolism, purine metabolism, amino acid metabolism, glycerophospholipid metabolism, and some other pathways. These findings provide a theoretical basis for the prevention and treatment of S. dysenteriae infection with extract from chestnut burs.

Medium-chain fatty acid production from thermal hydrolysed sludge without external electron donor supplementation.

In this study, medium-chain fatty acid (MCFA) generation from mixed sludge (including primary sludge and waste activated sludge) was investigated without additional electron donors (EDs). 0.5 g COD/L of MCFAs was produced and the in situ generated ethanol could serve as the EDs during the anaerobic fermentation of mixed sludge without thermal hydrolysis process (THP) pretreatment. THP increased the MCFA production by approximately 128% in the anaerobic fermentation. During 102 days of operation, the fermentation of THP pre-treated mixed sludge stably generated 2.9 g COD/L MCFAs. The self-generated EDs could not maximize MCFA production, and external addition of ethanol improved MCFA yield. Caproiciproducens was the dominant chain-elongating bacteria. PICRUST2 revealed that both fatty acid biosynthesis and reverse ß-oxidation pathways could participate in MCFA synthesis, and ethanol addition could enhance the contribution of the reverse ß-oxidation pathway. Future studies should focus on the improvement of MCFA production from THP-assisted sludge fermentation.

Bioactive constituents of animal-derived traditional Chinese medicinal materials for breast cancer: opportunities and challenges.

Breast cancer is globally the most common invasive cancer in women and remains one of the leading causes of cancer-related deaths. Surgery, radiotherapy, chemotherapy, immunotherapy, and endocrine therapy are currently the main treatments for this cancer type. However, some breast cancer patients are prone to drug resistance related to chemotherapy or immunotherapy, resulting in limited treatment efficacy. Consequently, traditional Chinese medicinal materials (TCMMs) as natural products have become an attractive source of novel drugs. In this review, we summarized the current knowledge on the active components of animal-derived TCMMs, including Ophiocordycepssinensis-derived cordycepin, the aqueous and ethanolic extracts of O.sinensis, norcantharidin (NCTD), Chansu, bee venom, deer antlers, Ostreagigas, and scorpion venom, with reference to marked anti-breast cancer effects due to regulating cell cycle arrest, proliferation, apoptosis, metastasis, and drug resistance. In future studies, the underlying mechanisms for the antitumor effects of these components need to be further investigated by utilizing multi-omics technologies. Furthermore, large-scale clinical trials are necessary to validate the efficacy of bioactive constituents alone or in combination with chemotherapeutic drugs for breast cancer treatment.

Preconcentration of liposoluble constituents in Salvia Miltiorrhiza using acid-assisted liquid phase microextraction based on a switchable deep eutectic solvent.

A switchable deep eutectic solvent-based liquid-phase microextraction was proposed and applied to the preconcentration and determination of liposoluble quality-markers of diterpenoid quinones (dihydrotanshinone I, cryptotanshinone, tanshinone I, and tanshinone IIA) in traditional Chinese medicine coupled with high performance liquid chromatography-ultraviolet detection. In the procedure, the hydrophilic deep eutectic solvent of diethanolamine-hexanoic acid (molar ratio 1:1) was prepared and added into the sample phase as an extractant, and a homogeneous solution was formed under slight vortex stirring. After the addition of HCl solution, the deep eutectic solvent miscible with the sample phase was converted to hydrophobic form, and a cloudy solution was generated. Then, the upper hydrophobic layer enriching the target analytes was collected through centrifugation for high performance liquid chromatography analysis. Several critical parameters affecting the extraction performance including the composition and consumption of switchable deep eutectic solvent, the type and amount of acid, salt amount and extraction time were investigated and optimized. Moreover, the structures of the deep eutectic solvent and the recovered hydrophobic layer were both characterized using Fourier transform infrared spectroscopy, further demonstrating the switching mechanism of the extractant during the extraction process. Under the optimal conditions, enrichment factors of diterpenoid quinones ranged from 59 to 274. Good linearities (r≥0.9963), low detection limits (0.5-0.7 ng/mL), satisfactory precisions (relative standard deviations 0.5%-8.6%) and accuracies (recoveries 94.6%-104.6%) were also obtained. Comparing the proposed switchable deep eutectic solvent-based liquid-phase microextraction with other published methods, the characteristics of the procedure were summarized. The developed method was successfully applied for the preconcentration of four liposoluble diterpenoid quinones from a traditional Chinese herbal medicine of Salvia Miltiorrhiza.

Effect of weimaining on apoptosis and Caspase-3 expression in a breast cancer mouse model.

ETHNOPHARMACOLOGICAL RELEVANCE: Weimaining (WMN) is a condensed Tannin compound extracted from Fagopyrum cymosum (Trevir.) Meisn., which comes from the roots of buckwheat, a type of Chinese herbal medicine, was first recorded in "Bencao Shiyi". WMN has inhibitory effects on multiple cancer types and is widely used in clinical practice; however, the mechanism underlying the anti-tumor effect of WMN is still unclear. AIM OF THE STUDY: To investigate the effect of WMN on the cellular activity and apoptosis of mouse breast cancer 4T1-luc2 cells, and caspase-3 and cleaved-caspase-3 expression. MATERIALS AND METHODS: Luciferase-labeled mouse breast cancer 4T1-luc2 cells were inoculated into the mouse breast pad to establish a luciferase-labeled mouse breast cancer cell model. BALB/C-nu mice were randomly divided into model, WMN, and low-molecular-weight heparin (LMWH) groups (n = 10). Another 10 mice served as the normal control group (no cancer cell injection). The WMN group was administered WMN 250 mg/kg per day for 14 days, the LMWH group was given LMWH (1500 U/kg) daily for 14 days by intraperitoneal injection, and the model and normal control groups were given an equal dose of 0.9% NaCl. The number and distribution of transplanted tumors in 4T1-luc2 breast cancer cells were observed in nude mice by an in vivo imaging system at the time of inoculation after successful modeling, and on days 7 and 14 after drug administration. Tumor cell apoptosis was detected by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method; caspase-3 mRNA expression was detected by RT-PCR and Western blotting was applied to detect the levels of caspase-3 and cleaved-caspase-3 protein expression. RESULTS: The apoptosis index (AI) of the WMN group was detected by the TUNEL method, and the AI increased with the increase of treatment time. Compared with the model group, the mRNA expression of caspase-3 and the protein levels of caspase-3 and cleaved-caspase-3 were notably elevated in the WMN group. After in vivo bioluminescent imaging, the total photon number of the WMN group was found to be lower than that of the LWMH group on day 14 after administration. Additionally, the AI and expression levels of caspase-3 mRNA, caspase-3, and cleaved-caspase-3 protein of the WMN group were higher than those of the LWMH group. CONCLUSION: WMN can effectively suppress the growth of 4T1-luc2 breast cancer xenografts in mice, and promote the apoptosis of breast cancer cells by upregulating the expression of caspase-3.

Compound Opening Arrow Mixture exerts anti-tumor effects in a mouse model of breast cancer.

Compound Opening Arrow Mixture (COAM) has demonstrated therapeutic effects in patients with breast cancer. We explored the underlying molecular mechanisms of COAM using a mouse model of breast cancer. Luciferase-labeled 4T1-Luc2 cells were inoculated into the breast pad of BALB/c-nu mice, which were divided into model group (saline), COAM (6 g/ml high-dose, 3 g/ml medium-dose, and 1.5 g/ml low-dose) groups, and low-molecular-weight heparin (LMWH, 1500 U/Kg) group. The number and distribution of 4T1-luc2 tumors were measured by an in vivo imaging system. Tumor cell apoptosis was measured through TUNEL and quantitating the expression of Caspase-3 mRNA and protein. Compared with the model group, in vivo tumor growth was lower in the LMWH- and COAM-treated groups. Tumor apoptosis was time-dependent and dose-dependent, as shown by a higher TUNEL apoptotic index and higher Caspase-3 mRNA and Caspase-3/cleaved-Caspase-3 proteins levels on the 14th day than the 7th day. The COAM high-dose group had the highest apoptotic index and the most activation of Caspase-3. Collectively, COAM significantly inhibits the growth of 4T1-luc2 breast cancer in mice and induces tumor apoptosis by activating Caspase-3, which provides a preliminary explanation of therapeutic effects of COAM.

Effects of Dangguibuxue decoction on rat glomerular mesangial cells cultured under high glucose conditions.

BACKGROUND: Dysfunction of glomerular mesangial cells (GMCs) plays an important role in pathogenesis of diabetic nephropathy. Here, we investigated the effects of Dangguibuxue decoction (DBD), an herbal traditional Chinese medicinal (TCM) formula composed of Astragali Radix and Angelicae Sinensis Radix, on GMC proliferation and fibrogenesis under high-glucose (HG) conditions. METHODS: Sixty male Sprague Dawley rats were divided into 5 groups and administered intragastric 0.9% saline, low concentration DBD (DBD-L, 1.75 g/kg/d), middle concentration DBD (DBD-M, 3.5 g/kg/d), high concentration DBD (DBD-H, 7.0 g/kg/d) and gliclazide (GL, 2 mg/kg/d), respectively, for 1 week, and then their sera were obtained. Rat mesangial cells (HBZY-1 cells) were treated with these sera under HG condition (30 mmol/L). RESULTS: The proliferation of GMCs under HG conditions was significantly greater than that under normal glucose condition. Low concentration DBD (DBD-L) inhibited proliferation of GMCs after 72-h incubation (P < 0.01), while high concentration DBD (DBD-H) inhibited GMCs proliferation at 24, 48 and 72 time points (P < 0.01). There was no significant difference between the inhibitory effect of DBD-H and GL sera on GMC proliferation (P > 0.05). Furthermore, all concentrations of DBD (DBD-L, DBD-M and DBD-H) significantly decreased the protein expression of α-SMA(α-smooth muscle actin) (P < 0.01), an indicator of interstitial fibrosis of GMCs. Finally, DBD-L, DBD-M, DBD-H sera obviously inhibited the increase of HYP (hydroxyproline)secretion under HG condition (P < 0.01). CONCLUSION: Our results demonstrate an inhibitory effect of DBD extract on proliferation and fibrogenesis of GMCs under HG conditions. The potential role of DBD in the treatment of diabetic neuropathy merits further investigation.

Simultaneous determination of phenolic compounds in sesame oil using LC-MS/MS combined with magnetic carboxylated multi-walled carbon nanotubes.

A novel magnetic carboxylated multi-walled carbon nanotubes (c-MWCNT-MNPs) was proposed for magnetic solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry to determine phenolic compounds in sesame oil. In this study, c-MWCNT-MNPs were acquired by simply dispersing Fe3O4 magnetic nanoparticles into carboxylated multi-walled carbon nanotubes. The major parameters affecting extraction efficiency were optimized, including the type and volume of desorption solvents, extraction and desorption time, washing solution, and sorbent amount. The limit of quantifications and limit of detections were from 0.03µg/kg to 43.00µg/kg and from 0.01µg/kg to 13.60µg/kg, respectively. The recoveries of phenolic compounds in vegetable oils were in the range of 83.8-125.9% with inter-day and intra-day precisions of less than 13.2%. It was confirmed that this method was simple, rapid and reliable with an excellent potential for routine analysis of phenolic compounds in oil samples.

Detection of virgin coconut oil adulteration with animal fats using quantitative cholesterol by GC × GC-TOF/MS analysis.

A new method based on the cholesterol level was developed to detect the presence of animal fats in virgin coconut oil (VCO). In this study, the sterols in VCO and animal fats was separated using conventional one-dimensional gas chromatography (1D GC) and comprehensive two-dimensional gas chromatography (GC×GC). Compared with 1D GC, the GC×GC system could obtain a complete baseline separation of the sterol trimethylsilyl ethers derived from cholesterol and cholestanol, so that the cholesterol content in pure VCO and false VCO adulterated with animal fats could be accurately determined. Cholesterol, a main sterol found in animal fats, represented less than 5mg/kg of VCO. The study demonstrated that the determination of the cholesterol level in VCO could be used for reliable detection of the presence of lard, chicken fat, mutton tallow, beef tallow, or their mixture in VCO at a level as little as 0.25%.

Simultaneous determination of seven ginsenosides in rat plasma by high-performance liquid chromatography coupled to time-of-flight mass spectrometry: application to pharmacokinetics of Shenfu injection.

A high-performance liquid chromatography coupled to time-of-flight mass spectrometry (HPLC-TOF MS) method was successfully developed and validated for the identification and determination of seven ginsenosides, Re , Rf , Rb1 , Rc , Rb2 , Ro and Rd , in a Chinese herbal preparation, Shenfu injection, and rat plasma. Based on the method, the pharmacokinetic profiles of the seven ginsenosides were investigated following intravenous administration of single dose of Shenfu injection to six rats. The established method had high linearity, selectivity, sensitivity, accuracy and precision. The pharmacokinetic results showed that Rb1 , Rc and Rb2 had similar pharmacokinetic profiles and relatively long half-life values (19.29 ± 6.36, 29.54 ± 22.91 and 35.60 ± 30.66 h). The half-lives of Rf and Rd were 4.21 ± 3.68 and 8.49 ± 5.20 h, respectively, indicating that they could be metabolized more rapidly than Rb1 , Rc and Rb2 .