Hypercalin B alleviates nonalcoholic steatohepatitis progression via suppressing mTORC1 signaling pathway / 药学学报

The global incidence rate of nonalcoholic steatohepatitis (NASH) continues to rise. The pathogenesis of NASH is complex, and there is no effective clinical treatment. Previous study has shown that DEAD box protein 5 (DDX5) can significantly alleviate the NASH process in mice. This study screened the natural product library of the research group and found that the active compound hypercalin B (HB) in Hypericum beanii N. Robson, a traditional Chinese medicine, can upregulate the expression of DDX5 protein in a dose-dependent manner. In this study, an in vitro model of NASH stimulated by palmitic acid (PA) and an animal model of NASH induced by the methionine- and choline-deficient diet (MCD) were constructed. Different concentrations of HB were used to investigate the effect and mechanism of HB in alleviating NASH progression. All animal experiments in this paper were approved by the Ethics Committee of China Pharmaceutical University (NO: 2021-02-003). In vitro model results showed that HB significantly reduced the intracellular lipid deposition induced by free fatty acid (FFA). Animal experiments showed that HB improved liver injury by significantly reducing lipid accumulation in the liver of NASH mice, and reducing serum aspartate transaminase (AST) and alanine transaminase (ALT) levels. Moreover, HB could inhibit liver inflammation by reducing the mRNA levels of liver pro-inflammatory cytokines including interleukin 6 (IL-6), interleukin 1β (IL-1β), and tumor necrosis factor α (TNFα). Further research showed that HB could reduce the phosphorylation level of the mechanical target of rapamycin (mTOR) and reduce the expression of sterol regulatory element binding protein 1 (SREBP1) and fatty acid synthase (FASN), thereby improving lipid metabolism and alleviating NASH progression, and the effects of HB against NASH were dependent on DDX5. In conclusion, HB can improve lipid metabolism and inhibit inflammatory activation by suppressing mTORC1 pathway via upregulating DDX5 protein, and showed promising anti-NASH activity in vitro and in vivo.

Root Rot Disease of Torreya grandis Caused by Fusarium fujikuroi in China.

Torreya grandis is an evergreen plant endemic of China and widely grown in Southern China. Its fruit is a precious nut in China, rich in vitamins and minerals, can be directly eaten, can also be used as medicinal plants with functions of lowering blood lipids and softening blood vessels (Wang 2022). From 2018 to 2020, typical root rot symptoms of Torreya grandis was found in plantations in Huangshan and surrounding areas of Huangshan, Anhui province, China. About 15 to 32% of root rot disease incidence was recorded at the plantation. Diseased plants were observed with symptoms such as yellow to brownish leaves without lesions and later drying, and rotten roots looked dark brown while the roots of heathy plants showed white, and eventually leading to the death of the diseased plant. The root rot symptomatic plants were collected in June of 2020. Tissues were cut to the length of 0.3 to 0.5 cm, then surface sterilized by 2% sodium hypochlorite for 2 min and 75% alcohol for 1 min, rinsed three times in sterile distilled water, and placed on potato dextrose agar (PDA) and incubated at 25℃ for 5 to 9 days. Eight isolates with similar morphology were isolated from single spores. On PDA, the isolates produced abundant aerial white mycelia with septation and turned violet to dark pink on the reverse side of the culture. Morphological characteristic was determined using a pure culture grown on synthetic low nutrient agar (SNA). Two types of conidia, microconidia and macroconidia, were observed on SNA. Macroconidia were long and slender, usually 3 to 5 septate, measuring 2.7 to 4.3 × 22.3 to 49.6 µm (n=30), and narrowed at the both ends. Microconidia were abundant, oval, clavate or ovate, zero to one septate and measured 1.6 to 3.9 × 4.4 to 13.0 µm (n=50). According to the culture and conidial characteristics, the isolates were tentatively identified as Fusarium species (Leslie and Summerell 2006). Four isolates were random selected for molecular identification. The general primers ITS1/ITS4 for internal transcribed spacer (ITS) (White et al. 1990), EF1/EF2 for translation elongation factor (TEF1) (O'Donnell et al. 1998), 5F2/7cR for the second largest subunit of RNA polymerase Ⅱ(RPB2) (O'Donnell et al., 2007), H3-1a/H3-1b for Histone H3 (Jacobs et al., 2010), F5/R8 for subunits 1 of DNA-directed RNA polymerase Ⅱ (RPB1) (O'Donnell et al. 2010) and MS3F/MS3R for mitochondrial small subunit (mtSSU) (Stenglein et al. 2010) were amplified, respectively. The products were sequenced and deposited in GenBank with accession numbers of MW350689, MW029444, ON077156, ON077158, ON077157, ON054432, respectively. Blast analysis showed 99.40 to 100% sequence homology with known F. fujikuroi isolates. A phylogenetic analysis based on the concatenated sequences clustered from the combined datasets (TEF1, RPB2, Histone H3, RPB1 and mtSSU) revealed the isolate most closely related to the F. fujikuroi (100% bootstrap). Fifteen 2-year-old healthy plants of Torreya grandis were selected for the pathogenicity test. A conidial suspension (1×106 conidia/ml) was prepared by collecting spores from 10-day-old cultures on PDA. The root of each plants inoculated with 200 ml of a 106 conidia/ml suspension, and the five control plants inoculated with sterilized water. The plants were incubated in green house with 25℃ (14 h light)/22℃ (10 h dark) at 85% humidity. Two weeks later, 100% of artificially inoculated plants showed the same symptoms similar to those observed in the plantation, like yellow leaves, dark brown and rotten roots, meanwhile, the roots of control plants displayed healthy. From symptomatic roots, the pathogen was reisolated which satisfying Koch's postulates. F. fujikuroi causes root rot of soybean and Reineckia carnea (Detranaltes et al. 2021, Sun et al. 2018).To the best of our knowledge, this is the first report of F. fujikuroi causing root rot of Torreya grandis in China.

Robust W/O/W Emulsion Stabilized by Genipin-Cross-Linked Sugar Beet Pectin-Bovine Serum Albumin Nanoparticles: Co-encapsulation of Betanin and Curcumin.

Betanin and curcumin hold promise as natural colorants and antioxidants for food purposes due to their anti-hypertensive, anti-inflammation, and anti-tumor effects. However, the thermal stability and bioavailability of betanin and curcumin still need improvement. Here, we fabricated sugar beet pectin-bovine serum albumin nanoparticles (SBNPs) with a mean particle size of 180 ± 5.2 nm through a genipin cross-linking strategy to stabilize a type of Pickering water-in-oil-in-water (W/O/W) emulsion and co-encapsulated betanin and curcumin. First, the W1/O emulsion was homogenized with gelatin (the gelling agent) in the water phase and polyglycerol polyricinoleate (a lipophilic surfactant) in the oil phase. Later, W1/O was homogenized with another water phase containing SBNPs. The microstructure of the emulsion was regulated by the particle concentration (c) and W1/O volume fraction (Φ), especially the gel-like high internal phase emulsions were formed at the Φ up to 70%. In this case, betanin was encapsulated in the internal water phase (encapsulation efficiency = 65.3%), whereas curcumin was in the medium-chain triglyceride (encapsulation efficiency = 84.1%). Meanwhile, the shelf stability of betanin and curcumin was improved. Furthermore, the stability of bioactive compounds was potentiated by an emulsion gel in simulated gastrointestinal digestion, resulting in higher bioaccessibility. The aforementioned results suggest that SBNP-stabilized Pickering W/O/W emulsions could be a potential alternative to co-encapsulate betanin and curcumin with enhancement of shelf stability and bioaccessibility.

New insights into the effects and mechanism of a classic traditional Chinese medicinal formula on influenza prevention.

BACKGROUND: KangBingDu (KBD) is a classic traditional Chinese medicinal formula widely used to treat influenza. However, little information is available from controlled studies regarding the anti-influenza pharmacological activities of KBD and its underlying mechanisms, at least partly due to the lack of appropriate study models. PURPOSE: We hypothesized that KBD might provide a protection against influenza infection by reducing the host's susceptibility to viruses. To prove it, mouse restraint stress model was employed. METHODS: Mice were restricted and infected with influenza virus. KBD (13 and 26mg/kg/d) was orally administrated to mice from the first day of restraint stress and lasted for 7 days (twice a day). Mice were monitored daily for morbidity, symptom severity, and mortality for 21 days. The histopathologic changes were examined. For the study of mechanisms of action, we investigated whether KBD could promote mitochondria antiviral signaling protein (MAVS)-mediated antiviral signal and inhibit nuclear factor-kappa B (NF-κB)-mediated inflammation response. RESULTS: KBD significantly decreased the susceptibility of restraint mice to influenza virus, as evidenced by lowered mortality, attenuated inflammation and reduced viral replications in lungs. Further results revealed that KBD elevated the protein expression of MAVS, which subsequently increased the IFN-ß and IFITM3 protein levels, thereby helping to fight viral infections. Finally, we identified that (R,S)-goitrin, mangiferin, forsythin and forsythoside A were effective components in KBD against influenza viral infections. CONCLUSION: KBD can reduce the susceptibility to influenza virus via mitochondrial antiviral signaling.

[Study on antimicrobial effect of the mixture of Chinese medicine-chitosan to Candida albicans and the possibility as denture adhesive].

PURPOSE: To observe the antimicrobial effect and the tensile bond strength of water-soluble chitosan after adding different Chinese medicines to Candida albicans. METHODS: The extract of 6 kinds of Chinese medicine by decoction in different concentrations were mixed with chitosan, and the most effective mixture inhibiting Candida albicans and the minimal inhibitory concentration (MIC) were explored; Then the tensile bond strength of the mixture was tested and compared with Protefix denture adhesive. The data was analyzed with SPSS17.0 software package. RESULTS: The antibacterial effect of polyphylla-chitosan mixture was the best among the 6 kinds of Chinese medicine- chitosan mixture, and its MIC was 1.563 mg/mL. The tensile bond strength of polyphylla-chitosan mixture at 0 h and 12 h when immersed in artificial saliva were significantly larger than Protefix denture adhesive. The average value and the maximum value was significantly greater than the Protefix denture adhesive (P<0.05). CONCLUSIONS: Polyphylla-chitosan mixture has good antibacterial effect on Candida albicans and large tensile bond strength.

Sesquiterpene coumarin and sesquiterpene chromone derivatives from Ferula ferulaeoides (Steud.) Korov.

Five novel compounds - three sesquiterpene coumarin derivatives, ferulin A (1), B (2), and C (3), and two sesquiterpene chromone derivatives, ferulin D (4) and E (5), together with eleven known compounds (6-16) have been isolated from the roots of Ferula ferulaeoides (Steud.) Korov. Their structures were established by comprehensive spectroscopic analysis. The biosynthetic pathways leading to these compounds were proposed. The cytotoxicity of all these isolates against HepG2, MCF-7, and C6 cancer cell lines was evaluated and compound 7 displayed the highest potency against HepG2, MCF-7, and C6 with IC50 values 39.9 µM, 37.7 µM, and 16.0 µM, respectively.

Sesquiterpenoid derivatives from Ferula ferulaeoides (Steud.) Korov.

Eight sesquiterpenoids, named Ferulaeone A-H (1-8), and seven known sesquiterpenoid derivatives were isolated from the roots of Ferula ferulaeoides (Steud.) Korov. Their structures were established by comprehensive spectroscopic analysis, and biosynthetic pathways leading to these compounds were proposed. The cytotoxicity of all these isolates against HepG2, MCF-7, and C6 cancer cell lines was evaluated and compounds 6-11, 13 exihibited various degrees of cytotoxic effect. Among them, compounds 9-11 displayed the highest potency against C6 with IC(50) values 34, 36, and 31 µM, respectively.

[Determination of vaccarin in Vaccariae Semen by HPLC].

To establish a HPLC method for the determination of vaccarin in Vaccariae Semen. Analysis was carried out on an Alltima-C18 column (4.6 mm x 250 mm, 5 microm) eluted with methanol -0.3% phosphoric acid as mobile phase in gradient elution. The flow rate was 1.0 mL x min(-1) and detected wavelength was set at 280 nm. The peak areas and injection ammounts of vaccarin showed a good linear relationship in the range of 0.102-1.539 microg, R2 = 0.9997. The average recovery was 100.4%, RSD was 0.81%. The results of the assay of 10 samples showed that the contents of vaccarin varied in the range of 0.46%-0.57%. The method is simple, accurate, reproducible and specific. It can be used for the quality control of Vaccariae Semen.

Y chromosomes of prehistoric people along the Yangtze River.

The ability to extract mitochondrial and nuclear DNA from ancient remains has enabled the study of ancient DNA, a legitimate field for over 20 years now. Recently, Y chromosome genotyping has begun to be applied to ancient DNA. The Y chromosome haplogroup in East Asia has since caught the attention of molecular anthropologists, as it is one of the most ethnic-related genetic markers of the region. In this paper, the Y chromosome haplogroup of DNA from ancient East Asians was examined, in order to genetically link them to modern populations. Fifty-six human remains were sampled from five archaeological sites, primarily along the Yangtze River. Strict criteria were followed to eliminate potential contamination. Five SNPs from the Y chromosome were successfully amplified from most of the samples, with at least 62.5% of the samples belonging to the O haplogroup, similar to the frequency for modern East Asian populations. A high frequency of O1 was found in Liangzhu Culture sites around the mouth of the Yangtze River, linking this culture to modern Austronesian and Daic populations. A rare haplogroup, O3d, was found at the Daxi site in the middle reaches of the Yangtze River, indicating that the Daxi people might be the ancestors of modern Hmong-Mien populations, which show only small traces of O3d today. Noticeable genetic segregation was observed among the prehistoric cultures, demonstrating the genetic foundation of the multiple origins of the Chinese Civilization.