Proteomics analysis of enzyme systems and pathway changes during the moromi fermentation of soy sauce mash.

BACKGROUND: During the brewing of soy sauce, the conversion of multiple substances is driven by various microorganisms and their secreted enzyme systems. Soy sauce mash is an important source of enzyme systems during moromi fermentation, but the changes of enzyme systems in soy sauce mash during moromi fermentation are poorly understood. In order to explore the predominant enzyme systems existing during moromi fermentation and to explain the characteristics of the enzyme system changes, an enzymatic activities assay and 4D-label-free proteomics analysis were conducted on soy sauce mash at different stages of fermentation. RESULTS: The activities of hydrolytic enzymes in soy sauce mash decreased continuously throughout the fermentation process, while most of the characteristic physicochemical substances in soy sauce mash supernatant had already accumulated at the early stage of fermentation. Four hydrolytic enzymes were found to be positively correlated with important physicochemical indexes by principal component analysis and Pearson correlation analysis. The proteomics analysis revealed three highly upregulated enzymes and two enzymes that were present in important metabolic pathways throughout the fermentation process. Furthermore, it was found that Aspergillus oryzae was able to accumulate various nutrients in the soy sauce mash by downregulating most of its metabolic pathways. CONCLUSION: Enzymes present with excellent properties during the moromi fermentation period could be obtained from these results. Meanwhile, the characterization of the metabolic pathways of microorganisms during the moromi fermentation period was revealed. The results provide a basis for more scientific and purposeful improvement of moromi fermentation in the future. © 2024 Society of Chemical Industry.

Interconvertible Pyridone Alkaloids from the Marine-Derived Fungus Penicillium oxalicum QDU1.

Eleven new pyridone alkaloids, penicipyridones A-K (1-11), and three new tetramic acids, tolypocladenols D-F (12-14), were isolated from rice media cultures of the marine-derived fungus Penicillium oxalicum QDU1. Their structures, including absolute configurations, were determined by comprehensive analyses of spectroscopic data, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction data. Interestingly, several of the penicipyridones undergo interconversions between hydroxy and methoxy groups at C-4 in acidic MeOH solution. Furthermore, in an acidic aqueous solution, OH-4 could be replaced by diverse substituent groups. Compounds 1, 4, 5, 8, 10, 11, and 14 exhibited moderate inhibitory effects on NO production in the LPS-induced RAW264.7 macrophages, with IC50 values ranging from 9.2 to 19 µM.

Naphtho-Gamma-Pyrones (NγPs) with Obvious Cholesterol Absorption Inhibitory Activity from the Marine-Derived Fungus Aspergillus niger S-48.

Eight naphtho-gamma-pyrones (NγPs) (1-8), together with four known biosynthetically related coumarin derivatives (9-12), were isolated from the potato dextrose agar media of a marine-derived fungus Aspergillus niger S-48. Among them, natural compounds 1 and 2 were tentatively subjected to benzohydrazide reaction to evaluate the importance of pyran rings in NγPs. Their structures were elucidated by extensive 1D and 2D NMR spectroscopic data and MS spectra. Compounds 1-4 showed obvious activity for reducing cholesterol absorption verging on ezetimibe. This work highlighted the potential of natural NγPs as NPC1L1 inhibitors.

[Efficacy of assisted treatment of thumb-tack acupuncture with surrounding needling method for herpes zoster of stagnated heat in liver meridian].

OBJECTIVE: To compare the efficacy difference between thumb-tack acupuncture with surrounding needling method plus medication and medication alone for herpes zoster (HZ) of stagnated heat in liver meridian type. METHODS: According to random number table method, 60 patients with HZ of stagnated heat in liver meridian type were randomly divided into an observation group and a control group, 30 cases in each one. All the patients were treated with oral valaciclovir hydrochloride dispersible tablets (0.3 g per time, twice per day), mecobalamin tablets (0.5 mg per time, three times a day) and vitamin B1 (10 mg per time, three times a day) for 15 days. In addition, the patients in the observation group were treated with thumb-tack acupuncture at area 1 cm outside the herpes, with an interval of 3 cm between thumb-tack needles. The thumb-tack acupuncture was given once every 3 days, retained for 48 h, with an interval of 1 day between treatments, and totally 5 treatments were given. The index of herpetic evaluation (stopping time of herpes, scarring time, decrustation time), visual analogue scale (VAS), serum immune-related factors (IgG, IgM, IgA) and serum inflammatory factors (IL-4, IL-17, TNF-α, TGF-ß1) were observed before and after treatment in the two groups. RESULTS: After treatment, the stopping time of herpes, scarring time, decrustation time in the observation group were shorter than those in the control group (all P<0.05). Compared before treatment, the VAS score in the two groups were reduced after treatment (both P<0.05), and no significant difference was observed between the two groups (P>0.05), but the difference before and after treatment in the observation group was superior to that in the control group (P<0.05). Compared before treatment, the levels of serum immune-related factors IgG, IgM, IgA were increased in the two groups after treatment (all P<0.05), and the levels in the observation group after treatment were higher than those in the control group (all P<0.05). Compared before treatment, the levels of serum inflammatory factors IL-4, IL-17, TNF-α, TGF-ß1 were reduced in the two groups after treatment (all P<0.05), and the levels in the observation group after treatment were lower than those in the control group (all P<0.05). CONCLUSION: The thumb-tack acupuncture with surrounding needling method plus medication have the advantages of rapid onset and analgesic effect for HZ of stagnated heat in liver meridian type, which could also improve serum immune-related factors and reduce inflammatory reaction.

Molecular identification of a root apical cell-specific and stress-responsive enhancer from an Arabidopsis enhancer trap line.

BACKGROUND: Plant root apex is the major part to direct the root growth and development by responding to various signals/cues from internal and soil environments. To study and understand root system biology particularly at a molecular and cellular level, an Arabidopsis T-DNA insertional enhancer trap line J3411 expressing reporters (GFP) only in the root tip was adopted in this study to isolate a DNA fragment. RESULTS: Using nested PCR, DNA sequencing and sequence homology search, the T-DNA insertion site(s) and its flanking genes were characterised in J3411 line. Subsequently, a 2000 bp plant DNA-fragment (Ertip1) upstream of the insert position of the coding T-DNA was in silico analysed, revealing certain putative promoter/enhancer cis-regulatory elements. Cloning and transformation of this DNA fragment and its truncated segments tagged with or without 35S minimal promoter (35Smini), all of which were fused with a GFP or GUS reporter, allowed to detect GFP and GUS expression mediated only by Ertip1 + 35mini (PErtip1+35Smini) specifically in the Arabidopsis root tip region. The PErtip1+35Smini activity was further tested to be strong and stable under many different growth conditions but suppressed by cold, salt, alkaline pH and higher ammonium and phosphorus. CONCLUSION: This work describes a promising strategy to isolate a tissue-/cell-specific enhancer sequence from the enhancer trap lines, which are publically available. The reported synthetic promoter i.e. PErtip1+35Smini may provide a valuable and potent molecular-tool for comprehensive investigation of a gene function related to root growth and development as well as molecular engineering of root-architectural formation aiming to improve plant growth.

[Discussion on needling manipulation of Chifeng Yingyuan].

Regarding the needling manipulation, Chifeng Yingyuan, there are different descriptions in ancient medical literature and the current teaching materials or clinical reports, especially for the understanding recorded in Jinzhenfu Poem of Golden Needle). In the current teaching materials and clinical reports, it is described that the needle is inserted deeply at first, and then shallowly; afterward, the needle tip is pushed to the middle layer and manipulated with lifting, thrusting and rotating technique, combined with twisting and flying method. After analyzing the original description in the book and the feasibility of function and operation, the authors introduced their recognition on its manipulation and techniques in this article. This manipulation should be: the needle is inserted to the middle layer, afterward, the needle body is pulled and tilted toward each of the four directions, on this basis, then twisting and flying method is applied.