Antitumor Effect of Poplar Propolis on Human Cutaneous Squamous Cell Carcinoma A431 Cells.

Propolis is a gelatinous substance processed by western worker bees from the resin of plant buds and mixed with the secretions of the maxillary glands and beeswax. Propolis has extensive biological activities and antitumor effects. There have been few reports about the antitumor effect of propolis against human cutaneous squamous cell carcinoma (CSCC) A431 cells and its potential mechanism. CCK-8 assays, label-free proteomics, RT-PCR, and a xenograft tumor model were employed to explore this possibility. The results showed that the inhibition rate of A431 cell proliferation by the ethanol extract of propolis (EEP) was dose-dependent, with an IC50 of 39.17 µg/mL. There were 193 differentially expressed proteins in the EEP group compared with the control group (p < 0.05), of which 103 proteins (53.37%) were upregulated, and 90 proteins (46.63%) were downregulated. The main three activated and suppressed Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were extracellular matrix (ECM)-receptor interaction, amoebiasis, cell adhesion molecules (CAMs), nonalcoholic fatty liver disease (NAFLD), retrograde endocannabinoid signaling, and Alzheimer's disease. The tumor volume of the 100 mg/kg EEP group was significantly different from that of the control group (p < 0.05). These results provide a theoretical basis for the potential treatment of human CSCC A431 cell tumors using propolis.

Pectic polysaccharides derived from Hainan Rauwolfia ameliorate NLR family pyrin domain-containing 3-mediated colonic epithelial cell pyroptosis in ulcerative colitis.

Pectic polysaccharides (PPs) could exert functions on ulcerative colitis (UC), which is classified as a nonspecific inflammatory disorder. This study investigated the molecular mechanism of PPs derived from Rauwolfia in UC. First, the dextran sodium sulfate (DSS)-induced mouse colitis models and lipopolysaccharide (LPS)-treated colonic epithelial cell (YAMC) models were established and treated with PP. Subsequently, the effects of PPs on mucosal damages in DSS mice were detected, and the levels of inflammatory cytokines, pyroptosis-related factors, oxidative stress-related markers, and the tight junction-related proteins in the tissues or cells were examined, and the results suggested that PPs ameliorated colonic mucosal damages and cell pyroptosis in DSS mice, and limited colonic epithelial cell pyroptosis in in vitro UC models. Subsequently, the binding relations of retinol-binding protein 4 (RBP4) to miR-124-3p and NLR pyrin domain-containing 3 (NLRP3) were analyzed. miR-124-3p targeted RBP4 and reduced the binding of RBP4 to NLRP3, thus inhibiting NLRP3-mediated pyroptosis. Finally, functional rescue experiments revealed that miR-124-3p suppression or RBP4 overexpression promoted colonic epithelial cell pyroptosis. Collectively, Rauwolfia-derived PPs limited miR-124-3p and targeted RBP4 and reduced the binding potency of RBP4 to NLRP3 to inhibit NLRP3-mediated pyroptosis, resulting in the alleviation of colonic epithelial cell pyroptosis and mucosal damages in UC.

Narciclasine ameliorated T cell mediated acute liver injury through activating AMPK pathway.

Hepatitis is closely related to cirrhosis and liver cancer, and it is vital that we develop new drugs and identify new drug targets. Traditional Chinese medicine has demonstrated excellent curative effects on liver diseases. The ingredients from Chinese herbals are important source for drug development in the treatment of hepatitis. Here, we found that narciclasine (NCS), a major component extracted from narcissus bulbs, showed hepatoprotective effect against concanavalin A (Con A) induced hepatitis. NCS treatment significantly reduced hepatocyte death, hepatic inflammatory cells infiltration, and serum cytokine levels in Con A challenged mice. We further observed that NCS directly inhibited Con A induced splenocytes proliferation and cytokine production in vitro. RNA-seq results showed that genes related to immune response were upregulated in Con A treated CD4+ T cells, which were down-regulated in the presence of NCS. Moreover, the AMPK pathway had been found activated in response to NCS treatment, suggesting a potential target for NCS targets. In conclusion, our results reveal that NCS is a powerful immunosuppressor against T cell activation, thus leading to protection against Con A induced liver injury in mice. These findings provide new insights into the use of natural products in the treatment of autoimmune hepatitis.

[Improving effect of Jiarong Tablets on the morphological structure and function of the testis in rats with late-onset hypogonadism].

OBJECTIVE: To investigate the effect of Jiarong Tablets (JRT) on the testicular morphology and function of rats with late-onset hypogonadism (LOH). METHODS: LOH models were established in 8 eighteen-month-old male SD rats, treated intragastrically with distilled water (the model control group, n = 4) or JRT at 0.375 g/kg/d, qd (the JRT group, n = 4), and another 5 two-month-old normal male SD rats were also given distilled water by gavage (normal control group), all for 28 days. Then all the rats were weighed and sacrificed for measurement of the serum T level and pathological and electron microscopic examination of the testis tissue. RESULTS: Compared with the normal controls, the LOH models showed significantly decreased testis coefficient (P < 0.05) and serum T level (ï¼»3.40 ± 0.06ï¼½ vs ï¼»5.88 ± 0.46ï¼½ ng /ml, P < 0.05). No statistically significant differences were observed in the model control and JRT groups in the body weight and testis coefficient (P > 0.05), but the serum T level (ï¼»4.50 ± 0.78ï¼½ ng/ml) was remarkably decreased in the latter (P < 0.05). In comparison with the model controls, the rats treated with JRT exhibited increases in the sperm count in the seminiferous tubules and the amount of testicular interstitial cells. Electron microscopy revealed a markedly increased number of mitochondria in the JRT-treated animals, with some mitochondrial sheaths and cristae but no obvious mitochondrial edema. CONCLUSIONS: Jiarong Tablets can elevate the serum T level and improve the testicular morphology and ultrastructure of LOH rats.

[Cloning and expression analysis of CtbHLH transcription factor gene from Carthamus tinctorius and construction of plant expression vector].

To clone bHLH( basic helix-loop-helix) gene from Carthamus tinctorius,analyze the expression level in different plant tissues and construct the plant expression vector. The bHLH1 gene was cloned by RT-PCR techniques,and the protein characteristics were analyzed by bioinformatics,and phylogenetic tree was constructed. The expression of bHLH1 gene in different tissues and the roots after inoculated by Fusarium oxysporum were analyzed using real time-PCR,and the plant expression vector p BASTA-bHLH1 was constructed. The obtained ORF sequence of bHLH1 gene was 897 bp,encoded a protein of 298 amino acids. Sequence alignment and phylogenetic tree analyses showed that C. tinctorius bHLH1 had a certain homology with other species of amino acids,and was the most similar to the amino acid sequence of tobacco. Real-time PCR results showed significant differences,CtbHLH1 gene in red flower petals in different tissues and different flowering period had remarkable difference in expression level,its high amount expressed in petals,flowers third day after blossom expressed the highest quantity,at the end of the flowering the expression quantity is low. In addition,it is expressed in the root,and the expression in the stem and leaves is extremely low. The bHLH1 gene of C. tinctorius is successfully cloned,and the expression is analyzed. The plant expression vector p BASTA-bHLH is constructed.

A UHPLC-MS/MS method for simultaneous determination of twelve constituents from Erigeron breviscapus extract in rat plasma: Application to a pharmacokinetic study.

A rapid, sensitive and specific ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated to simultaneously determine the twelve major bioactive ingredients (neochlorogenic acid, chlorogenic acid, caffeic acid, cynarin, scopoletin, scutellarin, isochlorogenic acid A, apigenin-7-o-glucuronide, isochlorogenic acid C, scutellarein, luteolin, and apigenin) in rat plasma. Gallic acid and wogonoside were used as internal standards (IS1 and IS2). The plasma samples were pretreated and extracted by liquid-liquid extraction and protein precipitation with ethyl acetate-acetonitrile (95:5, v/v). Chromatographic separation was accomplished on Agilent ZORBAX RRHD Eclipse Plus C18 column (2.1mm×50mm, 1.8µm) utilizing 0.1% formic acid aqueous solution and acetonitrile as mobile phase under gradient conditions at a flow rate of 0.3mL·min-1. Mass spectrometric detection was performed in multiple reaction monitoring (MRM) mode using electrospray ionization (ESI) in positive and negative mode. The whole intra- and inter-day precision (as relative standard deviation) of all analytes were less than 11.03%, and the accuracy (as relative error) were in the range from -10.43% to 9.76% and from -10.14% to 10.33%. The lower limits of quantification (LLOQ) were 20, 3.0, 100, 7.0, 0.30, 2.0, 70, 1.0, 20, 30, 10, and 2.0ngmL-1 for neochlorogenic acid, chlorogenic acid, caffeic acid, cynarin, scopoletin, scutellarin, isochlorogenic acid A, apigenin-7-o-glucuronide, isochlorogenic acid C, scutellarein, luteolin, and apigenin, respectively. Extraction recovery, matrix effect and stability were found to be the required limits. This method was selective and sensitive for the investigation of the pharmacokinetics of twelve constituents following oral administration to research study about in Erigeron breviscapus of clinical practices for separately analytes on rats.

Simultaneous determination and pharmacokinetic study of eight components in rat plasma by UHPLC-MS/MS after oral administration of Hypericum japonicum Thunb extract.

A rapid and sensitive assay based on ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was established and validated for the simultaneous determination of gallic acid, protocatechuic acid, vanillic acid, caffeic acid, epicatechin, isoquercitrin, vincetoxicoside B and quercetin in rat plasma using catechin and daidzein as the internal standards (IS). Plasma samples added internal standards were acidified with formic acid then pretreated by direct protein precipitation with acetonitrile. The separation of eight constituents was achieved on a C18 column with gradient elution using methanol and 0.2% acetic acid aqueous solution as the mobile phase and detected by multiple reaction monitoring using electrospray ionization source in the positive-negative ionization mode. The method was validated for sufficient specificity, precision, accuracy, and sensitivity over the concentration range of 10-6000 ng mL(-1) for gallic acid, 1.5-3000 ng mL(-1) for protocatechuic acid, 10-15000 ng mL(-1) for vanillic acid, 2-3600 ng mL(-1) for caffeic acid, 1.5-3600 ng mL(-1) for epicatechin, 4-6000 ng mL(-1) for isoquercitrin, 2-9000 ng mL(-1) for vincetoxicoside B, and 20-18000 ng mL(-1) for quercetin. The overall intra­run precision and the inter­run precision were showed in the range of 1.0-14.2% and 2.8-12.9%, respectively, and the accuracy was no more than 12.8%. This analytical method was successfully applied to investigate the pharmacokinetics of eight ingredients in rats after oral administration of Hypericum japonicum Thunb extract.

Comparative sucrose responsiveness in Apis mellifera and A. cerana foragers.

In the European honey bee, Apis mellifera, pollen foragers have a higher sucrose responsiveness than nectar foragers when tested using a proboscis extension response (PER) assay. In addition, Africanized honey bees have a higher sucrose responsiveness than European honey bees. Based on the biology of the Eastern honey bee, A. cerana, we hypothesized that A. cerana should also have a higher responsiveness to sucrose than A. mellifera. To test this hypothesis, we compared the sucrose thresholds of pollen foragers and nectar foragers in both A. cerana and A. mellifera in Fujian Province, China. Pollen foragers were more responsive to sucrose than nectar foragers in both species, consistent with previous studies. However, contrary to our hypothesis, A. mellifera was more responsive than A. cerana. We also demonstrated that this higher sucrose responsiveness in A. mellifera was not due to differences in the colony environment by co-fostering two species of bees in the same mixed-species colonies. Because A. mellifera foragers were more responsive to sucrose, we predicted that their nectar foragers should bring in less concentrated nectar compared to that of A. cerana. However, we found no differences between the two species. We conclude that A. cerana shows a different pattern in sucrose responsiveness from that of Africanized bees. There may be other mechanisms that enable A. cerana to perform well in areas with sparse nectar resources.