Transcriptome analysis revealing the effect of Bupleurum scorzonerifolium Willd association with endophytic fungi CHS3 on the production of saikosaponin D.

Saikosaponin D (SSd) is a naturally active product with strong pharmacological activity found in Bupleurum scorzonerifolium Willd. Studies have shown that endophytic fungi have great potential as sources of natural medicines. Fusarium acuminatum (CHS3), an SSd-producing endophytic fungus, was isolated from B. scorzonerifolium. To elucidate the effect of host plants on the production of SSd in CHS3, CHS3 was co-cultured with suspension cells of B. scorzonerifolium and SSd was detected using high-performance liquid chromatography (HPLC). Transcriptome sequencing (RNA-Seq) of CHS3 before and after co-culture was performed using an Illumina HiSeq 2500 platform. The results indicated that the content of SSd synthesised by CHS3 increased after co-culture with suspension cells of B. scorzonerifolium. Transcriptome analysis of CHS3 with differentially expressed genes (DEGs) showed that 1202 and 1049 genes were upregulated and downregulated, respectively, after co-culture. Thirty genes associated with SSd synthesis and 11 genes related to terpene backbone biosynthesis were annotated to the Kyoto Encyclopaedia of Genes and Genomes (KEGG). Combined with transcriptome data, it was speculated that the mevalonate (MVA) pathway is a possible pathway for SSd synthesis in CHS3, and the expression of key enzyme genes (HMGR, HMGCS, GGPS1, MVK, FDFT1, FNTB) was validated by qRT-PCR. In conclusion, the endophytic fungus CHS3 can form an interactive relationship with its host, thereby promoting SSd biosynthesis and accumulation by upregulating the expression of key enzyme genes in the biosynthesis pathway.

Concept and simulation of a novel dual-layer linear ion trap mass analyzer for micro-electromechanical systems mass spectrometry.

This paper proposed a dual-layer linear ion trap mass analyzer (dLIT) based on micro-electromechanical systems (MEMS) technology and stacked-layer structure for the development of MEMS mass spectrometry. Its basic performance and potential capabilities were explored by ion trajectory simulations. The theoretical formulas were modified by implementing multipole expansion. The simulation results were confirmed to be highly consistent with theoretical calculations in multiple aspects, including stability diagram, secular frequencies, and mass linearity, with only a deviation of 1-2%. In the boundary ejection mode, close to 100% ejection was achieved in a single dimension by applying extra quadrupole DC voltage. Preliminary simulation results showed that dLIT can achieve a peak width of ∼2 mass units (full width at half maximum, FWHM) for m/z 60 ions even at pressures as high as 50 Pa. Furthermore, the application of AC frequency scanning mode in dLIT was also evaluated, and preliminary simulation results yield a peak width of 0.3-0.4 mass units (FWHM). The dLIT offered several advantages, including high-precision fabrication at the sub-millimeter scale, excellent high-pressure performance, and a clear physical model. It preliminarily proved to be an ideal mass analyzer for MEMS mass spectrometry.

Sanghuangporus vaninii extract ameliorates hyperlipidemia in rats by mechanisms identified with transcriptome analysis.

The increasing incidence of hyperlipidemia is a serious threat to public health. The development of effective and safe lipid-lowering drugs with few side effects is necessary. The purpose of this study was to assess the lipid-lowering activity of Sanghuangporus vaninii extract (SVE) in rat experiments and reveal the molecular mechanism by transcriptome analysis. Hyperlipidemia was induced in the animals using a high-fat diet for 4 weeks. At the end of the 4th week, hyperlipidemic rats were assigned into two control groups (model and positive simvastatin control) and three treatment groups that received SVE at 200, 400, or 800 mg kg-1 day-1 for another 4 weeks. A last control group comprised normal chow-fed rats. At the end of the 8th week, rats were sacrificed and lipid serum levels, histopathology, and liver transcriptome profiles were determined. SVE was demonstrated to relieve the lipid disorder and improve histopathological liver changes in a dose-dependent manner. The transcriptomic analysis identified changes in hepatocyte gene activity for major pathways including steroid biosynthesis, bile secretion, cholesterol metabolism, AMPK signaling, thyroid hormone signaling, and glucagon signaling. The changed expression of crucial genes in the different groups was confirmed by qPCR. Collectively, this study revealed that SVE could relieve hyperlipidemia in rats, the molecular mechanism might be to promote the metabolism of lipids and the excretion of cholesterol, inhibit the biosynthesis of cholesterol by activating the AMPK signaling pathway, the thyroid hormone signaling pathway, and the glucagon signaling pathway.

Online water vapor removal membrane inlet mass spectrometer for high-sensitivity detection of dissolved methane.

Underwater mass spectrometry is characterized by excellent consistency, strong specificity, and the ability to simultaneously detect multiple substances, making it a valuable tool in research fields such as aquatic ecosystems, hydrothermal vents, and the global carbon cycle. Nevertheless, current underwater mass spectrometry encounters challenges stemming from the high-water vapor content, constituting proportions of nearly 90%. This results in issues such as peak overlap, interference with peak height, decreased ionization efficiency and, consequently, make it difficult to achieve low detection limits for extremely low concentrations of gases, such as methane, and impede the detection of background CH4 levels. In this study, we optimized the design of the sampling gas path and developed a high gas-tightness, high pressure-resistant membrane inlet system, coupled with a small-volume, low-power online water vapor removal system. This innovation efficiently eliminates water vapor while maintaining a high permeation flux of the target gases. By elevating the vacuum level to the order of 1E-6 Torr, the ionization efficiency and detection performance were improved. Based on this, we created an online water vapor removal membrane inlet mass spectrometer and conducted experimental research. Results indicated that the water removal efficiency approached 100%, and the vacuum level was elevated by more than 2 orders of magnitude. The detection limit for CH4 increased from over 600 nmol/L to 0.03 nmol/L, representing an improvement of over 4 orders of magnitude, and reaching the level of detecting background CH4 signals in deep-sea and lakes. Furthermore, the instrument exhibited excellent responsiveness and tracking capability to concentration changes on the second scale, enabling in situ analysis of rapidly changing concentration scenarios.

Transcriptome analysis of the mechanism of endophytic fungus CHS3 promoting saikosaponin d synthesis in Bupleurum scorzonerifolium Willd. suspension cells.

Saikosaponin d (SSd) is the main component of Bupleuri Radix, a famous traditional Chinese herbal medicine, with high medicinal value. An endophytic fungus (CHS3) was isolated from Bupleurum scorzonerifolium Willd. in the early stage of our research, and we found that CHS3 could promote the accumulation of SSd in Bupleurum scorzonerifolium Willd. suspension cells (BSS cells). It is of practical significance to identify the mechanism that CHS3 promoted the accumulation of SSd and increased the production of SSd in suspension cells. To search the influence of CHS3 on SSd synthesis in the BSS cells, we co-cultured CHS3 with the BSS cells and compared the SSd content in BSS cells before and after co-culture using high-performance liquid chromatography (HPLC). Then the Illumina HiSeq 2500 was performed to detect the transcriptome of the BSS cells before and after co-culture and analyzed for the KEGG enrichment. The expression of genes involved in SSd synthesis was finally corroborated by qPCR analysis. Among which 11 key genes in connection with SSd synthesis were increased in BSS cells of co-cultured group compared with the BSS cells of the control group. In conclusion, CHS3 could promote the accumulation of SSd in BSS cells, and the molecular mechanism was related to its ability to regulate the MVA pathway, the calcium signaling pathway, and the AMPK signaling pathway by upregulating the expressions of ANT, CypD, CaM, AMPK, AATC, HMGS, HMGR, MVK, MVD, SS, and SE.

Screening saikosaponin d (SSd)-producing endophytic fungi from Bupleurum scorzonerifolium Willd.

Saikosaponin d (SSd) is an important bioactive compound of traditional Chinese medicinal plant Bupleurum scorzonerifolium Willd. and exhibits many effects, such as anti-tumor, anti-inflammation and immunomodulatory. Since endophytic fungi possess the natural capacity to produce the similar secondary metabolite to that of their host plants, they are promising as alternative sources of plant bioactive natural products. In this study, in order to search for SSd-producing strains, endophytes were isolated from B. scorzonerifolium and were authenticated by the ITS sequence and the translation elongation factor-1alpha gene (TEF-1α) sequence analysis. The profile of metabolites present in the crude exacts was carried out by ultra performance liquid chromatography time-of-flight mass spectrometry (UPLC/Q-TOF-MS) analysis. The results showed that two strains, CHS2 and CHS3 from B. scorzonerifolium could produce SSd by UPLC/Q-TOF-MS analysis, and the amount of SSd produced by strain CHS2 and CHS3 were about 2.17 and 2.40 µg/mL, respectively. CHS2 and CHS3 showed a close phylogenetic relationship to Fusarium oxysporum and Fusarium acuminatum, respectively. According to our concern, no endophytic fungi capable of producing SSd from B. scorzonerifolium have been found before. Our clear intention was to isolate and identify these endophytic fungi that produce important active secondary metabolites, and then study the strains that produce this compound on a large scale through fermentation or even genetic study, to provide a feasible and more convenient way for the production of SSd.

Enhanced and sustainable pretreatment for bioconversion and extraction of resveratrol from peanut skin using ultrasound-assisted surfactant aqueous system with microbial consortia immobilized on cellulose.

In this study, the ultrasound-assisted surfactant aqueous system coupled with microbial consortia immobilized by cellulose has been created as an enhanced and sustainable method for the bioconversion and extraction of resveratrol from peanut skin. Based on central composite design, and several single-factor experiments, we derived the optimal bioconversion and extraction system. Microbial consortia consist of Yeast CICC 1912, Aspergillus oryzae 3.951 and Aspergillus niger 3.3148 were chosen to be immobilized using cellulose. Other treatment conditions include concentration of surfactant as 3% (w/v), temperature as 30 °C, time as 36 h, ultrasonic power as 250 W and liquid to solid ratio as 25:1 mL/g. Under these conditions, we achieved a promising yield of resveratrol 96.58 µg/g, which is 4.02 folds compared to the untreated sample. This sustainable and green method not only enhanced the production of resveratrol but also improved the safety and reliability of the bioconversion and extraction process. Our novel method has shown great potential to realize large-scale bioconversion and extraction of bioactive compounds from plant waste.

Protection of rat liver against hepatic ischemia-reperfusion injury by a novel selenocysteine-containing 7-mer peptide.

Hepatic ischemia-reperfusion (I-R) injury causes acute organ damage or dysfunction, and remains a problem for liver transplantation. In the I-R phase, the generation of reactive oxygen species aggravates the injury. In the current study, a novel selenocysteine-containing 7­mer peptide (H-Arg-Sec-Gly-Arg-Asn-Ala-Gln-OH) was constructed to imitate the active site of an antioxidant enzyme, glutathione peroxidase (GPX). The 7­mer peptide which has a lower molecular weight, and improved water­solubility, higher stability and improved cell membrane permeability compared with other GPX mimics. Its GPX activity reached 13 U/µmol, which was 13 times that of ebselen (a representative GPX mimic). The effect of this GPX mimic on I­R injury of the liver was assessed in rats. The 7­mer peptide significantly inhibited the increase in serum hepatic amino­transferases, tissue malondialdehyde, nitric oxide contents, myeloperoxidase activity and decrease of GPX activity compared with I­R tissue. Following treatment with the 7­mer peptide, the expression of B­cell CLL/lymphoma­2 (Bcl­2) was significantly upregulated at the mRNA and protein level compared with the I­R group, as determined by reverse transcription­polymerase chain reaction and immunohistochemistry, respectively. By contrast, Bcl­2 associated X protein (Bax) was downregulated by the 7­mer peptide compared the I­R group. Histological and ultrastructural changes of the rat liver tissue were also compared among the experimental groups. The results of the current study suggest that the 7­mer peptide protected the liver against hepatic I­R injury via suppression of oxygen­derived free radicals and regulation of Bcl­2 and Bax expression, which are involved in the apoptosis of liver cells. The findings of the present study will further the investigation of the 7-mer peptide as an effective therapeutic agent in hepatic I-R injury.

Lunasin suppresses the migration and invasion of breast cancer cells by inhibiting matrix metalloproteinase-2/-9 via the FAK/Akt/ERK and NF-κB signaling pathways.

Lunasin is a naturally existing bioactive peptide with an Arg-Gly-Asp (RGD) motif, which competes with integrins to bind with the extracellular matrix (ECM) consequently suppressing the integrin-mediated signaling pathway. Owing to the RGD motif, lunasin has been proven as an effective anti-inflammatory, antitumor and antimetastatic agent in many types of cancer. However, knowledge of its inhibitory effect on metastasis and the related mechanism of action in breast cancer cells is limited. In this study, the inhibitory effect of lunasin on the proliferation, migration and invasion of two typical breast cancer cell lines, ER-negative MDA-MB-231 with αVß3 expression and ER-positive MCF-7 with αVß5/α5ß1 expression, were examined in vitro as well the related mechanisms. The results demonstrated that lunasin (10-20 µM) effectively inhibited the migration and invasion activity and expression of matrix metalloproteinase (MMP)­2/-9 in both breast cancer cell lines. Meanwhile, we also found that lunasin inhibited the phosphorylation of focal adhesion kinase (FAK), Src, Akt, ERK and nucleus translocation of NF-κB, which indicates that, possibly via competing with αVß3 or αVß5/α5ß1 integrin, lunasin suppresses the metastasis of breast cancer cells through integrin-mediated FAK/Akt/ERK and NF-κB signaling pathways followed by downregulation of the activity and expression of MMP-2/-9.

[Study on Error Analysis of Nonlinear Function Coefficient of FAIMS].

The solution of ion mobility's nonlinear function coefficients α2 and α4 is the basis for achieving substance identification of High Field Asymmetric waveform Ion Mobility Spectrometry (FAIMS). Currently, nonlinear function coefficients α2 and α4 lack priors, meanwhile, existed solving results about α2 and α4 are deficient in error evaluation standard. In this article, acetone, isopropanol and 1, 2-dichlorobenzene were detected under different dispersion voltage by homemade FAIMS. In general, the spectrum peak of same sample at different dispersion voltage value is unique. Different dispersion voltage and corresponding compensation voltage value determines the value of α2 and α4. According to sample spectra at different dispersion voltage value, groups of spectral characteristics were obtained. Affirmatory number of data which were selected from multiple sets of compensation voltage value and dispersion voltage value, so that they were utilized to solved out lots of α2 and α4. Lots of factor have an effect on the accuracy of the solving results of α2 and α4, for instance, value of compensation voltage and dispersion voltage, style of fetching points of dispersion voltage, and so on. Comparing to other factors, style and amount of dispersion voltage is likely to control. By data analyzing huge amounts of α2 and α4 data, this paper explored their characteristic of distribution and correlation about them, research influence of number and method to fetch dispersion voltage detected points for error of solving results. After fitting frequency of α2 and α4, it was found that they conform to normal distribution, goodness of fitting exceed 0. 96, thus standard deviation of their distribution are able to evaluate error of solving results. In addition, a strong correlation exists between them, relevance of sample is -0. 977, -0. 968, -0. 992 respectively. With increasing of computing selected points, the corresponding error of solving results decrease. By comparing the standard deviation of method to fetch dispersion voltage detected points, found that detecting frequency in case of detecting maximum and the 70% of maximum of dispersion voltage value is lower at approximately same standard deviation, solving effect was optimized in unique fetching points style. Based on the premise of ensuring the accuracy of solving results of α2 and α4, it is obvious that reducing the frequency of detections for FAIMS effectively. It created favorable conditions for rapid field detection and precise spectral analysis.

Ameliorative Effects of Selenium on Cadmium-Induced Oxidative Stress and Endoplasmic Reticulum Stress in the Chicken Kidney.

The harmful influences of dietary cadmium (Cd) on the chicken kidney and the protective role of selenium (Se) against Cd-induced nephrotoxicity in the chicken are relatively unexplored subjects. The aim of this study was to investigate the ameliorative role of Se on the effects of Cd-induced oxidative stress, endoplasmic reticulum stress, and apoptosis in chicken kidneys. For this study, 100-day-old chickens received Se (as 10 mg Na2SeO3/kg dry weight of diet), Cd (as 150 mg CdCl2/kg dry weight of diet), or Cd + Se in their diets for 60 days. Then, the histopathological changes, Cd and Se contents, levels of oxidative stress, inducible nitric oxide synthase-nitric oxide (iNOS-NO) system activity, levels of endoplasmic reticulum (ER) stress, results of the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay of apoptosis, and expression levels of Bcl-2 and caspase 3 in the kidney were examined. The results showed that Cd exposure caused histopathological and ultrastructural damage and apoptosis of the kidneys. Cd administration significantly increased the accumulation of Cd, the malondialdehyde (MDA) content, NO production, iNOS activity, iNOS expression levels, expression levels of ER stress-related genes (GRP78, GRP94, ATF4, ATF6, and IRE) and the pro-apoptosis gene caspase 3, and the rate of apoptosis. Cd administration markedly decreased the Se content, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, and anti-apoptosis gene Bcl-2 expression levels. Co-treatment with Se and Cd obviously reduced the accumulation of Cd, Cd-induced histopathological and ultrastructural changes, oxidative stress, iNOS-NO system activity, ER stress, caspase 3 expression levels, and the rate of apoptosis in the kidneys. These results suggested that Cd exposure caused renal injury and that Se ameliorated Cd-induced nephrotoxicity in chickens.

[FAIMS of trace volatile organic compounds].

FAIMS is a fast and high sensitive technique for detecting trace volatile organic compounds. Spectra of acetone, benzene and toluene were obtained on a homemade high-field asymmetric waveform ion mobility spectrometer and they can be easily separated in the spectra. Three xylene isomeric compounds were also successfully separated in FAIMS. Effect of carrier gas flow rate on the ion intensity was analyzed, and the optimal flow rate of carrier gas was 220 L x h(-1) which can be used for the optimization of FAIMS instrument. The detection limit for acetone is 100 ng x L(-1) that is an order of magnitude lower than the foreign traditional IMS.