Mid-infrared spectroscopic identification of the right-baked rhubarb for ulcerative colitis therapy.

Clinical and experimental evidences have confirmed the significant therapeutic effects of rhubarb on ulcerative colitis (UC), but the strong purgative function of rhubarb also aggravates UC symptoms such as bloody diarrhea. Stir-baking to scorch is a traditional Chinese medicinal processing method that can eliminate the adverse purgative function while keep or even enhance the UC therapeutic function of rhubarb. However, the under-baked rhubarb still have the undesirable purgative function, but the over-baked rhubarb may lose the required medicinal functions. Therefore, the determination of the right endpoint is the primary quality concern about the baking process of rhubarb. In this research, typical anthraquinone compounds and mid-infrared (MIR) spectra were recruited to determine the best baking degree of rhubarb for UC therapy. Raw rhubarb slices were baked at 180 °C with rotation to prepare the rhubarbs with different baking degrees. The right-baked rhubarb was defined according to the UC therapeutic responses as well as the traditional color criterion. Referring to the typical anthraquinone compounds in rhubarb slices and extracts, the baking degree of rhubarb may be assessed by the conversion ratio of anthraquinone glycosides to anthraquinone aglycones. MIR spectra showed the gradual decompositions of organic compounds including anthraquinone glycosides and tannins during the baking process. Rhubarbs with different baking degrees can be distinguished clearly by MIR-based principal component analysis. In conclusion, the ratio of anthraquinone glycosides to anthraquinone aglycones may be a reasonable chemical indicator of the right-baked rhubarb. Meanwhile, MIR spectroscopy can identify the right-baked rhubarb simply and rapidly.

Ginsenoside Rg5 Activates the LKB1/AMPK/mTOR Signaling Pathway and Modifies the Gut Microbiota to Alleviate Nonalcoholic Fatty Liver Disease Induced by a High-Fat Diet.

The primary objective of this investigation was to elucidate the manner in which ginsenoside Rg5 (Rg5) ameliorates nonalcoholic fatty liver disease (NAFLD) via the modulation of the gut microbiota milieu. We administered either a standard diet (ND) or a high-fat diet (HFD), coupled with 12-week treatment employing two distinct doses of Rg5 (50 and 100 mg/kg/d), to male C57BL/6J mice. In comparison to the HFD cohort, the Rg5-treated group demonstrated significant enhancements in biochemical parameters, exemplified by a substantial decrease in lipid concentrations, as well as the reduced expression of markers indicative of oxidative stress and liver injury. This signifies a mitigation of hepatic dysfunction induced by an HFD. Simultaneously, Rg5 demonstrates the capacity to activate the LKB1/AMPK/mTOR signaling pathway, instigating energy metabolism and consequently hindering the progression of NAFLD. Furthermore, we underscored the role of Rg5 in the treatment of NAFLD within the gut-microbiota-liver axis. Analysis via 16S rRNA sequencing unveiled that Rg5 intervention induced alterations in gut microbiota composition, fostering an increase in beneficial bacteria, such as Bacteroides and Akkermansia, while concurrently reducing the relative abundance of detrimental bacteria, exemplified by Olsenella. Furthermore, employing fecal microbiota transplantation (FMT) experiments, we observed analogous outcomes in mice subjected to fecal bacterial transplants, providing additional verification of the capacity of Rg5 to mitigate NAFLD in mice by actively participating in the restoration of gut microbiota via FMT. Drawing from these data, the regulation of the gut microbiota is recognized as an innovative strategy for treating or preventing NAFLD and metabolic syndrome. Consequently, these research findings suggest that Rg5 holds promise as a potential therapeutic agent for NAFLD management.

One-Shot Single-Cell Proteome and Metabolome Analysis Strategy for the Same Single Cell.

Characterizing the profiles of proteome and metabolome at the single-cell level is of great significance in single-cell multiomic studies. Herein, we proposed a novel strategy called one-shot single-cell proteome and metabolome analysis (scPMA) to acquire the proteome and metabolome information in a single-cell individual in one injection of LC-MS/MS analysis. Based on the scPMA strategy, a total workflow was developed to achieve the single-cell capture, nanoliter-scale sample pretreatment, one-shot LC injection and separation of the enzyme-digested peptides and metabolites, and dual-zone MS/MS detection for proteome and metabolome profiling. Benefiting from the scPMA strategy, we realized dual-omic analysis of single tumor cells, including A549, HeLa, and HepG2 cells with 816, 578, and 293 protein groups and 72, 91, and 148 metabolites quantified on average. A single-cell perspective experiment for investigating the doxorubicin-induced antitumor effects in both the proteome and metabolome aspects was also performed.

The influence of ventilation modes on oil mist particles diffusion in a machining workshop.

Mechanical processing and operations are widely involved in modern industry. Large amount of oil mist is tended to be produced and will diffuse in the processing workshop when metalworking fluids are applied on the high temperature workpiece. The ventilation modes and air distributions can influence the air pollutants dilution in machining workshops. Therefore, this paper presents both experimental investigation and simulation study on the oil mist particles diffusion under different ventilation modes. The results identified PM2.5 as the primary component among different oil mist particles generated during a typical machining process. The distribution of oil mist particles in a full-scale machining workshop laboratory was investigated under two ventilation modes: high-sidewall nozzle air supply and low-sidewall air supply. Results revealed obvious influences of both air supply modes on the distribution of oil mist particles. Under the high-sidewall-nozzle air supply mode, the airflow and the oil mist distribution in the workshop was relatively uniform; while the low-sidewall-vent air supply mode led to an uneven distribution of oil mist particles, and the maximum oil mist concentration appeared at the height of 3 m. Under both modes, the attempts to increase the airflow rate are not always successful. Compared with low-sidewall-vent air supply mode, the high-sidewall-nozzle air supply mode presents better performance in achieving lower overall particle concentration level. Overall, the results of this study give useful reference to improve the air quality of industrial plant by properly designing the ventilation mode of machining workshop.

Pick-up single-cell proteomic analysis for quantifying up to 3000 proteins in a Mammalian cell.

The shotgun proteomic analysis is currently the most promising single-cell protein sequencing technology, however its identification level of ~1000 proteins per cell is still insufficient for practical applications. Here, we develop a pick-up single-cell proteomic analysis (PiSPA) workflow to achieve a deep identification capable of quantifying up to 3000 protein groups in a mammalian cell using the label-free quantitative method. The PiSPA workflow is specially established for single-cell samples mainly based on a nanoliter-scale microfluidic liquid handling robot, capable of achieving single-cell capture, pretreatment and injection under the pick-up operation strategy. Using this customized workflow with remarkable improvement in protein identification, 2449-3500, 2278-3257 and 1621-2904 protein groups are quantified in single A549 cells (n = 37), HeLa cells (n = 44) and U2OS cells (n = 27) under the DIA (MBR) mode, respectively. Benefiting from the flexible cell picking-up ability, we study HeLa cell migration at the single cell proteome level, demonstrating the potential in practical biological research from single-cell insight.

Two-dimensional correlation infrared spectroscopy elucidated the volatilization process of the microemulsion composed of peppermint essential oil and composite herbal extract.

Microemulsion is usually a transparent and isotropic liquid mixture composed of oil phase, water phase, surfactant and cosurfactant. The surfactant-framed nanoscale droplets in the microemulsion can penetrate into the skin surface to reduce its barrier function. This makes microemulsion an ideal preparation for the transdermal drug delivery. The permeability of microemulsion may be further enhanced when botanical essential oils that can dissolve the stratum corneum are used as the oil phase. However, the volatility of essential oils is possible to shorten the retention time of the microemulsion on the skin surface. Therefore, analytical methods are required to understand the volatilization process of the microemulsion composed of essential oils to develop the reasonable topical drug carrier system. In this research, Fourier transform infrared (FTIR) spectroscopy with an attenuated total reflection (ATR) accessory cooperated with two-dimensional correlation spectroscopy (2DCOS) to elucidate the volatilization processes of some microemulsions composed of peppermint essential oil. Principal component analysis (PCA) and moving-window two-dimensional correlation spectroscopy (MW2DCOS) revealed the multiple stages of the volatilization processes of the microemulsions. Synchronous 2D correlation infrared spectra indicated the compositional changes during each stage. It was found that the successive volatilizations of ethanol, water and menthone were the major events during the volatilization process of the microemulsion composed of peppermint essential oil. Ethanol can accelerate the volatilization of water, while the composite herbal extract seemed to not influence the volatilization of the other ingredients. After a 20-min-long volatilization process, the remaining microemulsion still contained considerable peppermint essential oil to affect the skin. The above results showed the feasibility of developing the microemulsion composed of peppermint essential oil for the transdermal drug delivery of composite herbal extract. This research also proved that the combination of ATR-FTIR spectroscopy and 2DCOS was valuable to study the volatilization process of the microemulsion.

CircMETTL15 induces TMTC3 production by absorbing miR-944 to promote hepatocellular carcinoma cell malignancy.

Previous data have suggested the involvement of circular RNA (circRNA) in hepatocellular carcinoma (HCC) progression. Up to now, the effect of circMETTL15 on HCC development remains unknown. This study aims to analyze the function of circMETTL15 in HCC development and the underlying mechanism. RNA expression of circMETTL15, miR-944, and transmembrane O-mannosyltransferase targeting cadherins 3 (TMTC3) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was evaluated by Western blot analysis assay or immunohistochemistry assay. Cell proliferation was investigated by cell counting kit-8 assay, 5-Ethynyl-29-deoxyuridine (EdU) assay, and cell colony formation assay. Cell migration and invasion were assessed by wound-healing assay and transwell assay, respectively. Angiogenic capacity was analyzed by tube formation assay. Dual-luciferase reporter assay and RNA immunoprecipitation assay were conducted to identify the interplay between miR-944 and circMETTL15 or TMTC3. Xenograft mouse model assay was conducted to reveal the effect of circMETTL15 on tumor formation in vivo. CircMETTL15 and TMTC3 expression were significantly upregulated, while miR-944 expression was downregulated in HCC tissues and cells. CircMETTL15 knockdown led to decreased cell proliferation, migration, invasion, and tube formation. Besides, the inhibitors of miR-944, a target miRNA of circMETTL15, partially restored circMETTL15 silencing-mediated effects on the proliferation, migration, invasion, and tube formation of HCC cells. MiR-944 overexpression also inhibited HCC cell malignancy by targeting TMTC3. Furthermore, circMETTL15 absence inhibited tumor formation by regulating miR-944 and TMTC3 in vivo. In conclusion, circMETTL15 induced HCC development through the miR-944/TMTC3 pathway, raising the potential of circMETTL15 as a target for HCC therapy.

Simultaneous deep transcriptome and proteome profiling in a single mouse oocyte.

Although single-cell multi-omics technologies are undergoing rapid development, simultaneous transcriptome and proteome analysis of a single-cell individual still faces great challenges. Here, we developed a single-cell simultaneous transcriptome and proteome (scSTAP) analysis platform based on microfluidics, high-throughput sequencing, and mass spectrometry technology to achieve deep and joint quantitative analysis of transcriptome and proteome at the single-cell level, providing an important resource for understanding the relationship between transcription and translation in cells. This platform was applied to analyze single mouse oocytes at different meiotic maturation stages, reaching an average quantification depth of 19,948 genes and 2,663 protein groups in single mouse oocytes. In particular, we analyzed the correlation of individual RNA and protein pairs, as well as the meiosis regulatory network with unprecedented depth, and identified 30 transcript-protein pairs as specific oocyte maturational signatures, which could be productive for exploring transcriptional and translational regulatory features during oocyte meiosis.

Structure characterization of novel heteropolysaccharides from Pteridium revolutum with antioxidant and antiglycated activities.

This study aims to analysis the structures of polysaccharides isolated from Pteridium revolutum and their antioxidant and antiglycated activities. Three novel water-soluble heteropolysaccharides, named PRP0, PRP1, and PRP2, were isolated from P. revolutum. The average molecular weight was determined by high performance gel permeation chromatography analysis as 1.04 × 106, 8.39 × 105, and 7.37 × 105 Da, respectively. Their structures were characterized using physicochemical and spectroscopic methods. The antioxidant and antiglycated activities were assayed in vitro. PRP0, PRP1, and PRP2 consist of l-Ara, l-Rha, d-Man, d-Xyl, d-Fuc, d-Gal, and d-Glc in different proportions. PRP1 mainly has a backbone of (1 â†’ 3,6)-linked d-Man and (1 â†’ 3)-linked d-Gal on main chain. PRP2 is mainly composed of (1 â†’ 2,4)-linked d-Man and (1 â†’ 3)-linked d-Gal on main chain. All polysaccharides have strong scavenging power on 2,2-difenil-1-picril-hidrazil and hydroxyl radicals and significantly antiglycated activity in Bovine serum albumin-Glucose model, which showing that the polysaccharides have potential application value on the functional food.

Phylogenetic relationship and characterization of the complete chloroplast genome of Rhododendron przewalskii subsp. przewalskii Maximowicz 1877, an ornamental and medicinal plant in China.

As one of the top 10 famous flowers in Chinese tradition, Rhododendron przewalskii subsp. przewalskii known as 'beauty in flowers,' which has high ornamental and medicinal value. The complete chloroplast (cp) genome of R. przewalskii subsp. przewalskii was determined in this study. The complete chloroplast genome of R. przewalskii subsp. przewalskii was 201,233 bp in length and contained a large single-copy region (LSC, 108,077 bp), and a small single-copy region (SSC, 2624 bp) and a pair of inverted repeat regions (IRa and IRb, 45,266 bp). A total of 142 functional genes were observed in this cp genome, including 91 protein-coding genes (PCGs), 43 transfer RNA genes (tRNAs), and eight ribosomal RNA genes (rRNAs). The R. przewalskii subsp. przewalskii cp genome has an A + T content of 64.06% and presents a positive AT-skew (0.53%) and a negative GC-skew (-1.56%). The maximum likelihood phylogenetic analysis based on the concatenated nucleotide sequences of 13 PCGs strongly supported the monophyletic relationship of R. przewalskii subsp. przewalskii the clade of R. henanense subsp. lingbaoense. This study provides genomic evidence for the vegetation classification of Rhododendron.

Ginsenoside Rg5 Improves Sleep by Regulating Energy Metabolism in Sleep-Deprived Rats.

Sleep deprivation (SD) has become a universal social problem. There is a causal relationship between SD and energy metabolism disorder. Phytochemicals have been demonstrated to have excellent sleep-promoting effects, and studies have shown that ginsenoside Rg5 (Rg5) exerts sedative and hypnotic effects. The present study aimed to investigate the role of Rg5 in regulating energy metabolism and explore the potential mechanism of improving sleep. Sleep-deprived rats were randomly divided into a control group (Ctrl), SD model group (SD), Rg5 group (GRg5), and melatonin group (MT). Sleep-deprived model rats were generated by housing rats in an SD box for 4 weeks. The Ctrl and SD groups were given equal volumes of saline. The Rg5 groups were given 25[Formula: see text]mg/kg Rg5 or 50[Formula: see text]mg/kg Rg5, and the MT group was given 0.27[Formula: see text]g/kg MT. A Western blot analysis and ELISA were used to detect the metabolic levels, mitochondrial functional proteins, AMPK pathway proteins, clock-related proteins, adenosine receptors, and neurotransmitter receptors. The results showed that Rg5 corrected abnormal glucose and lipid metabolism as well as improved ATP levels. In addition, Rg5 alleviated mitochondrial structural damage and improved the expression of proteins involved in mitochondrial biosynthesis, fission, and fusion. Moreover, Rg5 improved the expression of AMPK/PGC-1/Nrf-1 pathway proteins, regulated mitochondrial biological functions, and affected the rhythm characteristics of circadian clock-related proteins. Further, Rg5 improved the expression of A1R and A[Formula: see text]R as well as regulated the expression levels of GABAA1[Formula: see text] and mGluR5 to improve sleep in SD rats.

Chromatic Plasmonic Polarizer-Based Synapse for All-Optical Convolutional Neural Network.

Emerging memory devices have been demonstrated as artificial synapses for neural networks. However, the process of rewriting these synapses is often inefficient, in terms of hardware and energy usage. Herein, we present a novel surface plasmon resonance polarizer-based all-optical synapse for realizing convolutional filters and optical convolutional neural networks. The synaptic device comprises nanoscale crossed gold arrays with varying vertical and horizontal arms that respond strongly to the incident light's polarization angle. The presented synapse in an optical convolutional neural network achieved excellent performance in four different convolutional results for classifying the Modified National Institute of Standards and Technology (MNIST) handwritten digit data set. After training on 1,000 images, the network achieved a classification accuracy of over 98% when tested on a separate set of 10,000 images. This presents a promising approach for designing artificial neural networks with efficient hardware and energy consumption, low cost, and scalable fabrication.

Genomic profiling, prognosis, and potential interventional targets in young and old patients with cholangiocarcinoma.

Molecular mechanisms behind potentially inferior prognosis of old cholangiocarcinoma (CCA) patients are unclear. Prevalence of interventional targets and the difference between young and old CCA patients are valuable for promising precision medicine. A total of 188 CCA patients with baseline tumor tissue samples were subgrouped into the young (≤45 years) and old (>45 years) sub-cohorts. Somatic and germline mutation profiles, differentially enriched genetic alterations, and actionable genetic alterations were compared. An external dataset was used for the validation of molecular features and the comparison of overall survival (OS). Compared to young patients, KRAS alterations were more common in old patients (P = .04), while FGFR2 fusions were less frequent (P = .05). TERT promoter mutations were exclusively detected in old patients. The external dataset (N = 392) revealed no significant difference in OS between young and old patients; however, old patient-enriched KRAS (hazard ratio [HR]: 1.96, 95% confidence interval [CI]: 1.37-2.80) and TERT alterations (HR: 2.03, 95% CI: 1.22-3.38) were associated with inferior OS. Approximately 38.3% of patients were identified of actionable oncogenic mutations indicative of a potential response to targeted therapy or immunotherapy. Actionable FGFR2 fusions (P = .01) and BRAFV600E (P = .04) mutations were more frequent in young females than old patients. The enrichment of KRAS/TERT alterations in CCA patients over 45 years resulted in inferior OS. Approximately one-third of CCA patients were eligible for targeted therapy or immunotherapy given the actionable mutations carried, especially young females.

Study on the dynamic metabolic characteristic of main active ingredients in Danggui Buxue Decoction by liquid chromatography-tandem mass spectrometry based on in situ sequential metabolism strategy.

Danggui Buxue Decoction is a classic formula for replenishing qi and nourishing blood. Despite its widespread use, its dynamic metabolism involved remains unclear. Based on the sequential metabolism strategy, blood samples from different metabolic sites were obtained via in situ closed intestine ring integrated with a jugular venous continuous blood supply technique. An ultra-high-performance liquid chromatography-linear triple quadruple-Orbitrap-tandem mass spectrometry method was developed for the identification of prototypes and metabolites in rat plasma. The dynamic absorption and metabolic landscape of flavonoids, saponins, and phthalides were characterized. Flavonoids could be deglycosylated, deacetylated, demethylated, dehydroxylated, and glucuronicated in the gut and then absorbed for further metabolism. Jejunum is an important metabolic site for saponins biotransformation. Saponins that are substituted by Acetyl groups tend to lose their acetyl groups and convert to Astragaloside IV in the jejunum. Phthalides could be hydroxylated and glucuronidated in the gut and then absorbed for further metabolism. Seven components serve as crucial joints in the metabolic network and are potential candidates for the quality control of Danggui Buxue Decoction. The sequential metabolism strategy described in this study could be useful for characterizing the metabolic pathways of Chinese medicine and natural products in the digestive system.

A Novel Process for One-Step Separation and Cyclodextrin Inclusion of Ginsenoside Rg5 from Ginseng Stem-Leaf Saponins (GSLS): Preparation, Characterization, and Evaluation of Storage Stability and Bioactivity.

BACKGROUND: Ginsenoside Rg5 has been proven to possess numerous health benefits. However, Rg5 is difficult to prepare using the current methods, and the poor stability and solubility of Rg5 are intractable properties that limit its application. We try to establish and optimize a new method for preparing Rg5. METHODS: Different amino acids acted as catalysts, and reaction conditions were investigated to transform Rg5 in GSLS. Different CDs and reaction conditions were investigated for the preparation of CD-Rg5 based on yield and purity; ESI-MS, FT-IR, XRD and SEM analyses were used to prove the formation of the CD-Rg5 inclusion complex. Both the stability and bioactivity of ß-CD-Rg5 were investigated. RESULTS: The content of Rg5 reached 140.8 mg/g after transformation of GSLS using Asp as a catalyst. The yield of ß-CD-Rg5 reached a maximum of 12% and a purity of 92.5%. The results showed that the ß-CD-Rg5 inclusion complex can improve its stability of Rg5 against light and temperature. Antioxidant activity analyses against DPPH, ABTS+, and Fe2+ chelation showed enhanced antioxidant activity of the ß-CD-Rg5 inclusion complex. CONCLUSIONS: A novel and effective strategy for the separation of Rg5 from ginseng stem-leaf saponins (GSLS) was developed to improve the stability, solubility, and bioactivity of Rg5.

Promising Energetic Melt-Castable Material with Balanced Properties.

As one of the most widely used energetic materials to date, trinitrotoluene (TNT) suffers from several generally known drawbacks such as high toxicity, oil permeability, and poor mechanical properties, which are driving researchers to explore new high-performance energetic melt-castable materials for replacing TNT. However, it still remains a great challenge to discover a promising TNT alternative due to the multidimensional requirements for practical applications. Herein, we reported a new promising energetic melt-castable molecule, 4-methoxy-1-methyl-3,5-dinitro-1H-pyrazole (named as DMDNP). Besides a reasonable melting point (Tm: 94.8 °C), good thermostability (Td: 293.2 °C), and excellent chemical compatibility, DMDNP exhibits some obvious advantages over TNT including more environmentally friendly synthesis, high yield, low toxicity, low volume shrinkage, low mechanical and electrostatic sensitivities, etc., demonstrating well-balanced properties and great promise as a TNT replacement.

A transformer-based deep neural network model for SSVEP classification.

Steady-state visual evoked potential (SSVEP) is one of the most commonly used control signals in the brain-computer interface (BCI) systems. However, the conventional spatial filtering methods for SSVEP classification highly depend on the subject-specific calibration data. The need for the methods that can alleviate the demand for the calibration data becomes urgent. In recent years, developing the methods that can work in inter-subject scenario has become a promising new direction. As a popular deep learning model nowadays, Transformer has been used in EEG signal classification tasks owing to its excellent performance. Therefore, in this study, we proposed a deep learning model for SSVEP classification based on Transformer architecture in inter-subject scenario, termed as SSVEPformer, which was the first application of Transformer on the SSVEP classification. Inspired by previous studies, we adopted the complex spectrum features of SSVEP data as the model input, which could enable the model to simultaneously explore the spectral and spatial information for classification. Furthermore, to fully utilize the harmonic information, an extended SSVEPformer based on the filter bank technology (FB-SSVEPformer) was proposed to improve the classification performance. Experiments were conducted using two open datasets (Dataset 1: 10 subjects, 12 targets; Dataset 2: 35 subjects, 40 targets). The experimental results show that the proposed models could achieve better results in terms of classification accuracy and information transfer rate than other baseline methods. The proposed models validate the feasibility of deep learning models based on Transformer architecture for SSVEP data classification, and could serve as potential models to alleviate the calibration procedure in the practical application of SSVEP-based BCI systems.

Attenuated Total Reflection Fourier Transform Infrared Mapping for the Identification of the Prescribed and Abnormal Ingredients of Herbal Powder Preparations.

Herbal powder preparations (HPPs) are common forms of traditional medicine made by blending the powder of two or more ingredients. The first step to ensure the safety and efficacy of HPPs is to confirm the prescribed ingredients and screen the abnormal ingredients. With the help of attenuated total reflection Fourier transform infrared spectroscopy (ATR FT-IR) imaging or mapping, the particles of different ingredients in an HPP sample can be measured individually. In this way, the overlapped absorption signals of different ingredients in the ATR FT-IR spectrum of the bulk sample can be isolated in the ATR FT-IR spectra of the microscopic particles, which leads to the substantial increase of the specificity and sensitivity of the infrared spectral identification method. The characteristic particles of each ingredient can be identified by the objective comparison of the microscopic ATR FT-IR spectra against the reference spectra based on the correlation coefficients. Since the ATR FT-IR imaging or mapping tests of HPPs are free of the separation preprocess, multiple organic and inorganic ingredients are able to be recognized by a single identification procedure simultaneously rather than by different separation and identification procedures. As an example, the ATR FT-IR mapping method was used in this research to successfully identify three prescribed ingredients and two abnormal ingredients in oral ulcer pulvis, which is a classic HPP for oral ulcer in traditional Chinese medicine. The results show the feasibility of the ATR FT-IR microspectroscopic identification method for the objective and simultaneous identification of the prescribed and abnormal ingredients of HPPs.

Fourier transform infrared spectroscopic markers of the scorching degree of Scutellariae Radix related to the variation patterns of flavonoids and carbohydrates.

Scutellariae Radix (SR) is a common herb in Asia and Europe. In the clinical practice of traditional Chinese medicine, the raw SR is often stir-baked to partly scorch to reduce the side effects (stomach discomfort, diarrhea, etc.) but enhance some desired effects (such as the hemostatic activity). The scorching degree is crucial to assure the safety and efficacy of the scorched SR. Under-scorching is insufficient to adapt the adverse and favorable activities, while over-scorching can destroy all activities. Up to now, the scorching degree of SR is still determined by the manual observation of colors. Since the visual judgement is vulnerable to personal knowledge and experience, it is difficult to control the optimization and consistence of the scorching degree of SR. This research was designed to explore the potential indicators that can exactly reflect the scorching degree of SR and be measured objectively and quantitatively. A total of 15 morphological and chemical properties as well as Fourier transform infrared (FTIR) spectroscopy were included to profile the variations of the SR slices that were stir-baked at different temperatures for different times. According to the principal component analysis and statistical tests, 10 properties showed close relationships with the scorching degree of SR. As the increase of the heating time at the same heating temperature, the yield, L*, a*, b*, sucrose, baicalin and wogonoside decreased continuously and showed the monotone variation patterns. Meanwhile, 5-HMF, baicalein and wogonin showed the inflection variation patterns, which means these properties increased in the earlier stage but decreased in the later stage during the scorching process. FTIR spectra also revealed the variations of flavonoids and carbohydrates. The scorching degree of SR can be determined by the intuitive observation of FTIR spectral peaks at 1738 cm-1, 1705 cm-1, 1611 cm-1, 1586 cm-1, 1450 cm-1, 1410 cm-1 and 1023 cm-1. In summary, FTIR spectroscopy can be used for the rapid assessment of the scorching degree of SR, while the contents of flavonoid glycosides and aglycones may be used as the quality criteria of the scorched SR.

Medium optimization for high mycelial soluble protein content of Ophiocordyceps sinensis using response surface methodology.

Ophiocordyceps sinensis is widely utilized due to its pharmaceutical value. Mycelial protein forms a key active component of O. sinensis and determines the medicinal potential of fungus. Here, we describe the development of an optimized fermentation medium to obtain more mycelial soluble protein from O. sinensis using response surface methodology (RSM) and investigate the increased mycelial protein content using transcriptomics. The maximum mycelial protein content of 2.11% was obtained using a medium consisting of 20% beef broth, 0.10% peptone, 2% glucose, 0.15% yeast extract, 0.20% KH2PO4, and 0.02% MgSO4. Transcriptome analysis identified 790 differentially expressed genes (DEGs), including 592 up-regulated genes and 198 down-regulated genes, optimisation resulted in more up-regulated genes. The main DEGs were enriched in metabolic pathways, ABC transporters, starch and sucrose metabolism, tyrosine metabolism, and glutathione metabolism. In addition, some DEGs associated with mycelial protein enhancement such as tyrosinase (TYR), glutathione S-transferase (GST), glutamine synthetase (glnA), and ß-glucosidase may contribute to increased mycelial protein content. Real-time quantitative PCR (RT-qPCR) was used to confirm gene expression and the results support the accuracy of RNA-Seq and DEG analysis. This study provides an optimized fermentation method for enhancing the mycelial protein content of O. sinensis and a reference for the effective development of O. sinensis protein.