Facilitating Effects of Reductive Soil Disinfestation on Soil Health and Physiological Properties of Panax ginseng.

Chemical soil fumigation (CSF) and reductive soil disinfestation (RSD) have been proven to be effective agricultural strategies to improve soil quality, restructure microbial communities, and promote plant growth in soil degradation remediation. However, it is still unclear how RSD and CSF ensure soil and plant health by altering fungal communities. Field experiments were conducted to investigate the effects of CSF with chloropicrin, and RSD with animal feces on soil properties, fungal communities and functional composition, and plant physiological characteristics were evaluated. Results showed that RSD and CSF treatment improved soil properties, restructured fungal community composition and structure, enhanced fungal interactions and functions, and facilitated plant growth. There was a significant increase in OM, AN, and AP contents in the soil with both CSF and RSD treatments compared to CK. Meanwhile, compared with CK and CSF, RSD treatment significantly increased biocontrol Chaetomium relative abundance while reducing pathogenic Neonectria relative abundance, indicating that RSD has strong inhibition potential. Furthermore, the microbial network of RSD treatment was more complex and interconnected, and the functions of plant pathogens, and animal pathogen were decreased. Importantly, RSD treatment significantly increased plant SOD, CAT, POD activity, SP, Ca, Zn content, and decreased MDA, ABA, Mg, K, and Fe content. In summary, RSD treatment is more effective than CSF treatment, by stimulating the proliferation of probiotic communities to further enhance soil health and plant disease resistance.

Analysis Transcriptome and Phytohormone Changes Associated with the Allelopathic Effects of Ginseng Hairy Roots Induced by Different-Polarity Ginsenoside Components.

The allelopathic autotoxicity of ginsenosides is an important cause of continuous cropping obstacles in ginseng planting. There is no report on the potential molecular mechanism of the correlation between polarity of ginsenoside components and their allelopathic autotoxicity. This study applied a combination of metabolomics and transcriptomics analysis techniques, combined with apparent morphology, physiological indexes, and cell vitality detection of the ginseng hairy roots, through which the molecular mechanism of correlation between polarity and allelopathic autotoxicity of ginsenosides were comprehensively studied. The hairy roots of ginseng presented more severe cell apoptosis under the stress of low-polarity ginsenoside components (ZG70). ZG70 exerted allelopathic autotoxicity by regulating the key enzyme genes of cis-zeatin (cZ) synthesis pathway, indole-3-acetic acid (IAA) synthesis pathway, and jasmonates (JAs) signaling transduction pathway. The common pathway for high-polarity ginsenoside components (ZG50) and ZG70 to induce the development of allelopathic autotoxicity was through the expression of key enzymes in the gibberellin (GA) signal transduction pathway, thereby inhibiting the growth of ginseng hairy roots. cZ, indole-3-acetamid (IAM), gibberellin A1 (GA1), and jasmonoyl-L-isoleucine (JA-ILE) were the key response factors in this process. It could be concluded that the polarity of ginsenoside components were negatively correlated with their allelopathic autotoxicity.

A Ratiometric Fluorescence Detection Method for Berberine Using Triplex-Containing DNA-Templated Silver Nanoclusters.

Berberine (BBR), as a natural isoquinoline alkaloid, has demonstrated various pharmacological activities, and is widely applied in the treatment of diseases. The quantitative analysis of BBR is important for pharmacological studies and clinical applications. In this work, utilizing the specific interaction between BBR and triplex DNA, a sensitive and selective fluorescent detecting method was established with DNA-templated silver nanoclusters (DNA-AgNCs). After binding with the triplex structure in the template of DNA-AgNCs, BBR quenched the fluorescence of DNA-AgNCs and formed BBR-triplex complex with yellow-green fluorescence. The ratiometric fluorescence signal showed a linear relationship with BBR concentration in a range from 10 nM to 1000 nM, with a detection limit of 10 nM. Our method exhibited excellent sensitivity and selectivity, and was further applied in BBR detection in real samples.

Analysis of the Variation in Antioxidant Activity and Chemical Composition upon the Repeated Thermal Treatment of the By-Product of the Red Ginseng Manufacturing Process.

Steamed ginseng water (SGW) is a by-product of the repeated thermal processing of red ginseng, which is characterized by a high bioactive content, better skin care activity, and a large output. However, its value has been ignored, resulting in environmental pollution and resource waste. In this study, UHPLC-Q-Exactive-MS/MS liquid chromatography-mass spectrometry and multivariate statistical analysis were conducted to characterize the compositional features of the repeated thermal-treated SGW. The antioxidant activity (DPPH, ABTS, FRAP, and OH) and chemical composition (total sugars, total saponins, and reducing and non-reducing sugars) were comprehensively evaluated based on the entropy weighting method. Four comparison groups (groups 1 and 3, groups 1 and 5, groups 1 and 7, and groups 1 and 9) were screened for 37 important common difference markers using OPLS-DA analysis. The entropy weight method was used to analyze the weights of the indicators; the seventh SGW sample was reported to have a significant weight. The results of this study suggest that heat treatment time and frequency can be an important indicator value for the quality control of SGW cycling operations, which have great potential in antioxidant products.

Comprehensive Theoretical Study of Cp*IrIII-Catalyzed Intermolecular Enantioselective Allylic C-H Amidation: Reaction Mechanism, Electronic Processes, and Regioselectivity.

Density functional theory was used to elucidate the reaction mechanism of Cp*IrIII-catalyzed intermolecular regioselective C(sp3)-H amidation of alkenes with methyl dioxazolones. All substrates, intermediates, and transition states were fully optimized at the ωB97XD/6-31G(d,p) level (LANL2DZ(f) for Ir). The computational results revealed that this amidation occurred through the IrIII/IrV catalytic cycle, involving four important elementary steps: C-H bond activation, oxidative addition of methyl dioxazolone, reductive elimination, and proto-demetalation, and the first was the rate-determining step. The C-H bond activation showed good α- and branch-regioselectivity, decided by the distortion energy of 2-pentene and the interaction energy of the transition state, respectively. The oxidative addition of dioxazolone occurred in one elementary step with CO2 disassociation. The reductive elimination showed good branch-regioselectivity determined by the distorted energy of the allyl group. In the proto-demetalation, hydrogen directly transferred from the oxygen atom to the nitrogen atom. Moreover, to clarify the effect of the substituted groups, selected 12 substrates were also discussed in this text.

Carbon nanodots constructed by ginsenosides and their high inhibitory effect on neuroblastoma.

BACKGROUND: Neuroblastoma is one of the common extracranial tumors in children (infants to 2 years), accounting for 8 ~ 10% of all malignant tumors. Few special drugs have been used for clinical treatment currently. RESULTS: In this work, herbal extract ginsenosides were used to synthesize fluorescent ginsenosides carbon nanodots via a one-step hydrothermal method. At a low cocultured concentration (50 µg·mL- 1) of ginsenosides carbon nanodots, the inhibition rate and apoptosis rate of SH-SY5Y cells reached ~ 45.00% and ~ 59.66%. The in vivo experiments showed tumor volume and weight of mice in ginsenosides carbon nanodots group were ~ 49.81% and ~ 34.14% to mice in model group. Since ginsenosides were used as sole reactant, ginsenosides carbon nanodots showed low toxicity and good animal response. CONCLUSION: Low-cost ginsenosides carbon nanodots as a new type of nanomedicine with good curative effect and little toxicity show application prospects for clinical treatment of neuroblastoma. It is proposed a new design for nanomedicine based on bioactive carbon nanodots, which used natural bioactive molecules as sole source.

First Report of Pantoea agglomerans Causing Leaf Blight on Schisandra chinensis in China.

Schisandra chinensis (Turcz.) Baill is a perennial liana, which is widely cultivated and used in China. In August 2022, Schisandra chinensis leaves with small light brown spots were found on plants growing in Fusong (127°28'E, 42°33'N) of China. There was 15% disease incidence and 50% disease severity of Schisandra chinensis in 2-ha fields of S. chinensis. As the disease progressed, the spots become darker and form round or irregular concentric circles. Leaves with brown spot symptoms were collected from the field. Leaf pieces (5 mm × 5 mm) were excised from lesion margins, surface disinfected with 75% ethanol for 1 min, followed by 1.5% sodium hypochlorite for 3 min, and incubated on Luria Bertani (LB) solid medium at 28°C for 24 hours. Eight cultures were isolated, and representative single colony (XWWZH) was selected from the pure cultures according to colony characteristics for observation The purified colonies were round, yellow, and slimy, cells were straight rod-shaped (0.40 to 0.52 × 1.12 to 1.69 µm) were observed. The isolate was Gram negative. It was positive for methyl red reaction, lysine decarboxylase reaction, gelatin hydrolysis reactionand sucrose utilization. It was negative for indole reaction and produced H2S. The bacterium was preliminarily identified as Pantoea agglomerans based on morphological and biochemical tests (Baird et al. 2007). The 16S rDNA and a portion of rpoB of strain XWWZH were amplified and sequenced. The sequences were submitted to GenBank. (Accession OP763753 and OQ813505, respectively). Phylogenetic trees were constructed based on the 16S rDNA and rpoB gene sequences. The sequences of strain XWWZH clustered with strains P. agglomerans deposited in GenBank. The pathogenicity was verified with non-wounded S. chinensis seedlings by punching holes with sterile needles and injecting a solution of 1 × 108 CFU/ml solution. Sterile ddH2O was injected in the control experiment. The inoculated seedlings were incubated in a greenhouse at 25°C with a relative humidity of 65 to 70%. Five to eight days after inoculation, inoculated leaves, exhibited symptoms which were morphologically identical to those of the originally infected leaves whereas control plants remained asymptomatic. The pathogenicity assays were repeated twice with the same results. The re-isolated pathogen had the same morphology and DNA sequences as the original isolate obtained from the field samples, completing Koch's postulates. Strains of P. agglomerans have been reported to severely infect many plants (Ren et al.2008; Lee et al. 2010; Yang et al. 2011; Guo et al. 2019; Gao et al, 2022), but to the best of our knowledge, this is the first report of a strain of P. agglomerans causing leaf blight on Schisandra chinensis in China. The identification of leaf blight caused by P. agglomerans will enable farmers to prevent and manage it ahead of time to reduce losses.

First Report of Fusarium acuminatum Causing Leaf Spot on Schisandra chinensis in China.

Schisandra chinensis (Turcz.) Baill. is a popular and widely cultivated medicinal herb in China, which has rich nutritional value and medicinal effect. In August 2022, leaves with oval and irregularly circular light brown spots from 2 to 10 mm wide with white centers were found on Schisandra chinensis growing in Fusong district (127°28'E, 42°33'N) of Jilin, China. The symptoms were observed in 20% of the plants of a 2 ha-1 field of Schisandra chinensis. About 50% of the leaf areas were affected. As the disease developed, the lesions grew larger and developed necrotic centers. Leaves with light brown spot symptoms from five plants were collected from the field. Five leaf pieces (3 to 5 mm2) were excised from lesion margins, surface sterilized based on Ju et al. (Ju et al. 2021), and incubated on potato dextrose agar (PDA) at 25°C. Six single spores were isolated from five independently infected isolates for pure culture using the single spore isolation technique (Zhang. 2003). Representative single spore isolate (ZWWZH) was selected from pure cultures for further culture. After 5 days, fluffy white aerial mycelium with pink pigmentation on the underside of the colony were observed on PDA. Mycelia became pinkish-brown as the culture aged. Microscopic observations showed the presence of elongated or pointed, and thick-walled macroconidia (n = 50), predominantly three septate, 3.40 to 7.50 × 40.34 to 61.29 µm were observed. Chlamydospores formed in chains within or on top of the mycelium. The primers ITS1/ITS4 (White et al. 1990) and Bt-2a/Bt-2b (Robideau et al., 2011) were used to amplify the internal transcribed spacer (ITS) rDNA and ß-tubulin (TUB2) region, respectively. The obtained sequences were submitted to GenBank under accession numbers for OQ629789 (ITS) and OQ803521(TUB2). BLASTn analysis of both ITS sequence and TUB2 sequence, revealed 100% and 99.92% sequence identity with F. acuminatum MT566456, MT560377 and KJ396328, respectively. The pathogen was identified as F. acuminatum based on morphological and molecular data. Pathogenicity tests were carried out in the greenhouse. Select five healthy Schisandra chinensis seedlings, each with each healthy leaf surfaces inoculated a 1 × 106 spores/mL solution, 3 wells on one side, 10 µL per well. Sterile ddH2O was used in the control experiment. The inoculated seedlings were incubated at 25°C with a relative humidity of 65 to 70% in a greenhouse. Four days after inoculation, all inoculated leaves exhibited the same symptoms as observed in the field, while the controls showed no symptoms. The experiment was repeated three more times with similar results. The re-isolated fungi from the inoculated plants had the same morphology and DNA sequences as the original isolate (ZWWZH) obtained from the field samples, completing Koch's postulates. To our knowledge, this is the first report of F. acuminatum causing leaf spot on Schisandra chinensis in China. F. acuminatum has seriously affected the quality of Schisandra chinensis production. The identification of leaf spot caused by F. acuminatum will enable farmers to identify practices to minimize disease on this important crop.

Detection of catecholamine metabolites in urine based on ultra-high-performance liquid chromatography-tandem mass spectrometry.

The excretion of neurotransmitter metabolites in normal individuals is of great significance for health monitoring. A rapid quantitative method was developed with ultra-performance liquid chromatography-tandem mass spectrometry. The method was further applied to determine catecholamine metabolites vanilymandelic acid (VMA), methoxy hydroxyphenyl glycol (MHPG), dihydroxy-phenyl acetic acid (DOPAC), and homovanillic acid (HVA) in the urine. The urine was collected from six healthy volunteers (20-22 years old) for 10 consecutive days. It was precolumn derivatized with dansyl chloride. Subsequently, the sample was analyzed using triple quadrupole mass spectrometry with an electrospray ion in positive and multireaction monitoring modes. The method was sensitive and repeatable with the recoveries 92.7-104.30%, limits of detection (LODs) 0.01-0.05 µg/mL, and coefficients no less than 0.9938. The excretion content of four target compounds in random urine samples was 0.20 ± 0.086 µg/mL (MHPG), 1.27 ± 1.24 µg/mL (VMA), 3.29 ± 1.36 µg/mL (HVA), and 1.13 ± 1.07 µg/mL (DOPAC). In the urine, the content of VMA, the metabolite of norepinephrine and adrenaline, was more than MHPG, and the content of HVA, the metabolite of dopamine, was more than DOPAC. This paper detected the levels of catecholamine metabolites and summarized the characteristics of excretion using random urine samples, which could provide valuable information for clinical practice.

By-Product of the Red Ginseng Manufacturing Process as Potential Material for Use as Cosmetics: Chemical Profiling and In Vitro Antioxidant and Whitening Activities.

Red ginseng (RG), which is obtained from heated Panax ginseng and is produced by steaming followed by drying, is a valuable herb in Asian countries. Steamed ginseng dew (SGD) is a by-product produced in processing red ginseng. In the present study, phytochemical profiling of extracts of red ginseng and steamed ginseng dew was carried out using gas chromatography-mass spectrometry (GC-MS) and rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (RRLC-Q-TOF-MS) analysis. Additionally, antioxidant activities (DPPH, ·OH, and ABTS scavenging ability) and whitening activities (tyrosinase and elastase inhibitory activity) were analyzed. Phytochemical profiling revealed the presence of 66 and 28 compounds that were non-saponin components in chloroform extracts of red ginseng and steamed ginseng dew (RG-CE and SGD-CE), respectively. Meanwhile, there were 20 ginsenosides identified in n-butanol extracts of red ginseng and steamed ginseng dew (RG-NBE and SGD-NBE). By comparing the different polar extracts of red ginseng and steamed ginseng dew, it was found that the ethyl acetate extract of red ginseng (RG-EAE) had the best antioxidant capacity and whitening effect, the water extract of steamed ginseng dew (SGD-WE) had stronger antioxidant capacity, and the SGD-NBE and SGD-CE had a better whitening effect. This study shows that RG and SGD have tremendous potential to be used in the cosmetic industries.

Non-targeted metabonomics to investigate the differences in the properties of ginseng and American ginseng based on rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry.

The basic properties of herbal medicines are cold, hot, warm, and cool. The differentiation of these properties is important for the diagnosis and treatment of diseases. Ginseng and American ginseng possess opposite properties of warm and cool, respectively. At present, the mechanisms and the influence of steaming leading to the differences in their properties are not clear and require further investigation. Therefore, nontargeted metabonomics based on liquid chromatography-mass spectrometry was applied to investigate the effects of ginseng, American ginseng, and their variants on the changes in endogenous metabolites in rat urine. A total of 19 potential biomarkers were screened out and identified, of which 17, 7, and 5, were respectively related to warm, cool, and both warm and cool properties with opposite effects. The metabolic pathways corresponded to fatty acids, lipids, glycolysis, and energy metabolisms. The warm and tonic effects of red ginseng are stronger than those of ginseng and consistent with the theory of traditional Chinese medicine. The red American ginseng has cool property; however, the degree of coolness is less than that of American ginseng. This study provides a reference methodology to understand the effects of processing and mechanisms associated with the differences in the properties of herbal medicines.

Preparation, Characterization, and Bioavailability of Host-Guest Inclusion Complex of Ginsenoside Re with Gamma-Cyclodextrin.

This work aimed at improving the water solubility of Ginsenoside (G)-Re by forming an inclusion complex. The solubility parameters of G-Re in alpha (α), beta (ß), and gamma (γ) cyclodextrin (CD) were investigated. The phase solubility profiles were all classified as AL-type that indicated the 1:1 stoichiometric relationship with the stability constants Ks which were 22 M-1 (α-CD), 612 M-1 (ß-CD), and 14,410 M-1 (γ-CD), respectively. Molecular docking studies confirmed the results of phase solubility with the binding energy of -4.7 (α-CD), -5.10 (ß-CD), and -6.70 (γ-CD) kcal/mol, respectively. The inclusion complex (IC) of G-Re was prepared with γ-CD via the water-stirring method followed by freeze-drying. The successful preparation of IC was confirmed by powder X-ray diffraction (XRD), Fourier transform-infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), and scanning electron microscopy (SEM). In-vivo absorption studies were carried out by LC-MS/MS. Dissolution rate of G-Re was increased 9.27 times after inclusion, and the peak blood concentration was 2.7-fold higher than that of pure G-Re powder. The relative bioavailability calculated from the ratio of Area under the curve AUC0-∞ of the inclusion to pure G-Re powder was 171%. This study offers the first report that describes G-Re's inclusion into γ-CD, and explored the inclusion complex's mechanism at the molecular level. The results indicated that the solubility could be significantly improved as well as the bioavailability, implying γ-CD was a very suitable inclusion host for complex preparation of G-Re.

Investigation of the Effects of Squalene and Squalene Epoxides on the Homeostasis of Coenzyme Q10 in Rats by UPLC-Orbitrap MS.

Squalene has been used as a dietary supplement for a long history due to its potential cancer-preventive function. However, the mechanism has not been investigated in detail yet. Therefore, the aim of this study is to see if the plasma coenzyme Q10 (CoQ10) level will be altered by gavage of squalene and oxidosqualenes to rats. In the present work, a sensitive and simple high-performance analytical method based on ultra-high-performance liquid chromatography coupled with an Orbitrap mass spectrometry (UPLC-Orbitrap-MS) was developed for the quantification of CoQ10 in rat plasma. Coenzyme Q9 (CoQ9) was employed as the internal standard. CoQ10 was determined after acetonitrile-mediated plasma protein precipitation using UPLC-Orbitrap-MS in negative ion mode. Intragastric administration of squalene and the two squalene epoxides into rats once daily for several days elevated the level of CoQ10 in their plasma, but there was no significant difference between high-dose (286 mg/kg) and low-dose (143 mg/kg) groups. Intragastric administration of squalene once a day for 5 consecutive days and oxidosqualenes once a day for 3 consecutive days is necessary for reaching the steady-state level of CoQ10. Our present findings indicate that squalene and oxidosqualenes may be useful for stimulating the synthesis of CoQ10 in rats.

PRMT1 potentiates chondrosarcoma development through activation of YAP activity.

Protein arginine methyltransferase 1 (PRMT1) is identified as an oncogene implicated in various types of human cancers, while Yes-associated protein (YAP) as a key transcriptional coactivator of the Hippo signaling plays a vital role in tissue homeostasis and tumorigenesis. To date, the underlying biological functions, prognostic values, and potential mechanisms of PRMT1 and YAP in chondrosarcoma development have not been clearly elucidated. Here, we show that upregulation of PRMT1 and YAP is significantly detected in human chondrosarcoma specimens. Elevated levels of PRMT1 positively correlated with YAP nuclear accumulation are significantly associated with high-grade chondrosarcoma and poor prognosis. Moreover, YAP is recognized as an independent prognostic factor for chondrosarcoma patients. Ectopic expression of PRMT1 potentiates, but depletion of PRMT1 attenuates, chondrosarcoma cell growth in vitro and in vivo. Mechanistically, we have discovered that PRMT1 functions upstream of LATS1 and suppresses LATS1-mediated phosphorylation of YAP (Ser127), and thus promotes chondrosarcoma cell survival in a YAP-dependent manner. Collectively, our study identifies PRMT1 as a positive regulator of YAP activity in chondrosarcoma, highlighting a novel therapeutic target against chondrosarcoma and other YAP-driven cancers.

Magnetic silica nanoparticles for use in matrix-assisted laser desorption ionization mass spectrometry of labile biomolecules such as oligosaccharides, amino acids, peptides and nucleosides.

Magnetic silica nanoparticles (MSNPs) were prepared and applied for the first time as a matrix in MALDI MS for analysis of small thermally labile biomolecules including oligosaccharides, amino acids, peptides, nucleosides, and ginsenosides. The matrix was characterized by scanning electron microscopy and UV-vis spectroscopy. It displays good performance in analyses of such biomolecules in the positive ion mode. In addition, the method generates significantly less energetic ions compared to the use of carbon nanotubes or graphene-assisted LDI MS and thus produces intact molecular ions with little or no fragmentation. In addition, the MSNPs have better surface homogeneity and better salt tolerance and cause lower noise. It is assumed that the soft ionization observed when using MSNPs as a matrix is due to the specific surface area and the homogenous surface without large clusters. The matrices were applied to the unambiguous identification and relative quantitation of the water extract of Panax ginseng roots. Any false-positive results as obtained when using graphene and carbon nanotubes as a matrix were not observed. Graphical abstract Schematic presentation of the application of magnetic silica nanoparticles in laser desorption ionization mass spectrometry. Their use results in little or no fragmentation during analysis of small labile biomolecules with some advantages such as better surface homogeneity, high salt tolerance, and lower noise.

Screening and investigation of triplex DNA binders from Stephania tetrandra S. Moore by a combination of peak area-fading ultra high-performance liquid chromatography with orbitrap mass spectrometry and optical spectroscopies.

The identification and screening of triplex DNA binders are important because these compounds, in many cases, are potential anticancer agents as well as promising drug candidates. Therefore, the ability to screen for these compounds in a high-throughput mode could dramatically improve the drug screening process. A method involving a combination of 96-well plate format and peak area-fading ultra high-performance liquid chromatography coupled with Orbitrap mass spectrometry was employed for screening bioactive compounds binding to the triplex DNA from the extracts of Stephania tetrandra S. Moore. Two compounds were screened out and identified as fangchinoline and tetrandrine based on the comparison of retention time and tandem mass spectrometry data with those of standards. The binding mechanisms of fangchinoline and tetrandrine at the molecular level were explored using tandem mass spectrometry, fluorescence spectroscopy, ultraviolet-visible spectroscopy, and circular dichroism. Collision-induced dissociation experiments showed that the complexes with fangchinoline and tetrandrine were dissociated by ligand elimination. According to these measurements, an intercalating binding is the most appropriate binding mode of these two alkaloids to the triplex DNA. The current work provides not only deep insight into alkaloid-triplex DNA complexes but also useful guidelines for the design of efficient anticancer agents.

High-throughput screening of triplex DNA binders from complicated samples by 96-well pate format in conjunction with peak area-fading UHPLC-Orbitrap MS.

Conventional strategies for the screening of DNA triplex binders cannot be used for complicated samples, such as ligand libraries created by combinatorial chemistry or from natural product extracts. In the current study, an ultra-high-performance liquid chromatography coupled with an Orbitrap mass spectrometry (UHPLC-Orbitrap-MS)-based approach, which we call peak area-fading (PAF) UHPLC-Orbitrap-MS and was designed for just such a purpose, is reported. The triplex DNA modified 96-well plate and the single stranded oligonucleotide modified 96-well plate (as control) were incubated with ligand libraries, and the unbound ligands were directly determined via UHPLC-ESI-MS. The binders were detected through the decrease (fading) in the peak areas compared to those of the control group. Several factors, such as incubation time, incubation temperature, and buffer, which might affect the binding affinity and reproducibility, were optimized. The potential of the approach was examined using the extracts of Rhizoma Coptidis and Phellodendron chinense Schneid cortexe. The triplex DNA-binding capabilities of the five components (epiberberine, coptisine, jatrorrhizine, berberrubine, and columbamine) were found for the first time, indicating their efficiency for the analysis of complicated samples. In contrast to our previous study, which suffered from a serious drawback of poor reproducibility, this method is more robust and more suitable for high-throughput measurements, opening a new experimental strategy in assessing large libraries of potential drug candidates that work by forming a drug/DNA complex.

Posterior short-segment instrumentation and limited segmental decompression supplemented with vertebroplasty with calcium sulphate and intermediate screws for thoracolumbar burst fractures.

PURPOSE: Thoracolumbar burst fractures treated with short-segment posterior instrumentation without anterior column support is associated with a high incidence of implant failure and correction loss. This study was designed to evaluate the clinical and radiographic results following posterior short-segment instrumentation and limited segmental decompression supplemented with vertebroplasty with calcium sulphate and intermediate screws for patients with severe thoracolumbar burst fractures. METHODS: Twenty-eight patients with thoracolumbar burst fractures of LSC point 7 or more underwent this procedure. The average follow-up was 27.5 months. Demographic data, radiographic parameters, neurologic function, clinical outcomes and treatment-related complications were prospectively evaluated. RESULTS: Loss of vertebral body height and segmental kyphosis was 55.3 % and 20.2° before surgery, which significantly improved to 12.2 % and 5.4° at the final follow-up, respectively. Loss of kyphosis correction was 2.2°. The preoperative canal encroachment was 49 % that significantly improved to 8.8 %. The preoperative pain and function level showed a mean VAS score of 9.2 and ODI of 89.9 % that improved to 1.4 and 12.9 % at the final follow-up, respectively. No implant failure was observed in this series, and cement leakage occurred in two cases without clinical implications. CONCLUSIONS: Excellent reduction and maintenance of thoracolumbar burst fractures can be achieved with short-segment pedicle instrumentation supplemented with anterior column reconstruction and intermediate screws. The resultant circumferential stabilization combined with a limited segmental decompression resulted in improved neurologic function and satisfactory clinical outcomes, with a low incidence of implant failure and progressive deformity.

The influence of storage conditions on the quality of Schisandra chinensis fruits: A integrated investigation of constituent and antioxidant activity.

Schisandra chinensis is a functional fruit with tonic effect and was widely used by traditional Chinese medicine for treatment and health care. The quality of Schisandra chinensis fruit may vary by different storage condition. In this study, the influence of ambient temperature, humidity, packaging materials and period of storage on the quality of Schisandra chinensis fruits were investigated. The contents of main active components lignans and organic acids were simultaneously determined by ultra-performance liquid chromatography coupled to quadrupole electrostatic field orbitrap high resolution mass spectrometry (UPLC Orbitrap HRMS). The antioxidant activity was determined using DPPH radical scavenging capacity, ABTS+ inhibition rate and FRAP value. The correlation of multicomponent and antioxidant activity was analyzed by grey relevance analysis. Taking the changes of multicomponent and antioxidant activity as investigation index, Schisandra chinensis fruits under different storage conditions was comprehensively evaluated. Schisandrol A, malic acid, sorbic acid, schizandrin A, schizandrin B, and schisandrol B were the main effective components of antioxidant activity. Ambient temperature at 5 °C and humidity at 40 % were more suitable for Schisandra chinensis fruits and kraft paper bag was better packaging material. Do not exceed 1 year was the effective storage period. For the safety evaluation, no aflatoxin was detected within the storage period of 2 years, demonstrated the storage was satisfactory. This study provided a reference for the high-quality storage and standardized operating procedures for storage of Schisandra chinensis fruits.

6,7-Dimethoxycoumarin, Gardenoside and Rhein combination improves non-alcoholic fatty liver disease in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: This study explores the potential therapeutic benefits of using a three-component DGR (composed of specific compounds) to target the NLRP3 inflammasome in the context of non-alcoholic fatty liver disease (NAFLD). AIM OF THE STUDY: To assess the impact of a three-component DGR on NAFLD, specifically examining its effects on liver lipid accumulation, inflammation, and the diversity of intestinal microbial communities. METHODS: NAFLD was induced in 8-week-old Sprague Dawley rats by feeding them a high-fat emulsion diet every morning for 8 consecutive weeks. Oral administration of DGR or its constituent equivalents in the afternoon. The pharmacological effects of DGR were evaluated using H&E, ORO and ELISA methods to determine the changes in serum and liver tissue indexes of rat-models. Immunohistochemical staining and Western blot were used to assess the interaction between DGR, NLRP3 and IL-1ß. RESULTS: The induction of NAFLD resulted in elevated hepatic triglycerides (TG), total cholesterol (TC), and free fatty acids (FFA). However, these alterations were ameliorated upon administration of DGR. It is noteworthy that DGR exhibited superior efficacy in comparison to its constituent compounds, manifesting augmented antioxidant activity, diminished hepatic damage, and the attenuation of pro-inflammatory factors. Both DGR and its individual monomeric constituents exhibited the capacity to attenuate the activation of the NLRP3 inflammasome in the liver, leading to an amelioration of the pathological characteristics associated with NAFLD. An analysis of the intestinal flora unveiled an elevated abundance of p_Firmicutes (1.1-fold), p_Cyanobacteria (5.76-fold), and p_Verrucomicrobia (5.2-fold), accompanied by a heightened p_Firmicutes to p_Bacteroidetes ratio (5.49-fold). CONCLUSIONS: In the non-alcoholic fatty liver disease (NAFLD) rat model, the concurrent administration of three-component DGR effectively regulated lipid deposition, suppressed liver inflammation, and restored balance in the intestinal flora, thereby improving NAFLD pathology. These findings propose a promising therapeutic strategy for NAFLD, centered on inhibiting the NLRP3 inflammasome through the use of the three-component DGR.