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Over the past decade, advancements in high-throughput sequencing technologies have led to a qualitative leap in our understanding of the role of the microbiota in human diseases, particularly in oncology. Despite the low biomass of the intratumoral microbiota, it remains a crucial component of the tumor immune microenvironment, displaying significant heterogeneity across different tumor tissues and individual patients. Although immunotherapy has emerged a major strategy for treating tumors, patient responses to these treatments vary widely. Increasing evidence suggests that interactions between the intratumoral microbiota and the immune system can modulate host tumor immune responses, thereby influencing the effectiveness of immunotherapy. Therefore, it is critical to gain a deep understanding of how the intratumoral microbiota shapes and regulates the tumor immune microenvironment. Here, we summarize the latest advancements on the role of the intratumoral microbiota in cancer immunity, exploring the potential mechanisms through which immune functions are influenced by intratumoral microbiota within and outside the gut barrier. We also discuss the impact of the intratumoral microbiota on the response to cancer immunotherapy and its clinical applications, highlighting future research directions and challenges in this field. We anticipate that the valuable insights into the interactions between cancer immunity and the intratumoral microbiota provided in this review will foster the development of microbiota-based tumor therapies.

Abstract Head and neck squamous cell carcinoma (HNSCC) is a common malignant tumour. Despite advancements in surgery, radiotherapy and chemotherapy, which have improved the prognosis of most patients, a subset of patients with poor prognoses still exist due to loss of surgical opportunities, postoperative recurrence, and metastasis, among other reasons. The tumour microenvironment (TME) is a complex organization composed of tumour, stromal, and endothelial cells. Communication and interaction between tumours and immune cells within the TME are increasingly being recognized as pivotal in inhibiting or promoting tumour development. Previous studies on T cells in the TME of HNSCC have yielded novel therapeutic possibilities. However, the function of B cells, another adaptive immune cell type, in the TME of HNSCC patients has yet to be determined. Recent studies have revealed various distinct subtypes of B cells and tertiary lymphoid structures (TLSs) in the TME of HNSCC patients, which are believed to impact the efficacy of immune checkpoint inhibitors (ICIs). Therefore, this paper focuses on B cells in the TME to explore potential directions for future immunotherapy for HNSCC.

The urban (suburban) railway is a fast and convenient rail transit system connecting urban and suburban areas, and a refined analysis of the diversity of public service facilities around its stations can help to promote the intensive use of land around rail stations. However, the differences in the diversity of public service facilities in the railway life circle between urban and suburban railway stations and the factors affecting them are not clear. This paper takes the Beijing Suburban Railway Line Sub-center (Line S1) as a case study, uses the Shannon-Wiener index to measure the spatial diversity characteristics of public service facilities, and utilizes a multi-scale geographically weighted regression model to explore the influencing factors. The findings indicate that: (1) Centered on the stations, all six stations show a “less-more-less” ring or half-ring to the left distribution structure of the comprehensive diversity index of public service facilities within their study areas, with an increase followed by a decrease. (2) The influence of each influencing factor on the diversity of market-featured facilities exhibits significant differences. The most substantial spatial heterogeneity is observed in the distances to the nearest subway stations and bus stops. Distances to subway and urban (suburban) railway stations exhibit different spatial distribution characteristics within urban and suburban areas on Line S1. In urban areas, the closer the distance to the subway station or the further the distance to the railway station, the greater the diversity of public service facilities. Conversely, in suburban areas, the opposite is true. The conclusions of this research provide a scientific methodology and improvement measures to facilitate the construction of railway life circles in suburban regions of megacities.

Abstract A low sidelobe 77 GHz centre‐fed microstrip patch array antenna is proposed for automotive radar application. The proposed antenna comprises eight linear antenna arrays, each of which is centre‐fed by a coplanar waveguide with ground transmission line at the back of the microstrip patches. Each linear antenna array contains 10 patch antenna units, which are connected through a T‐type power divider network to achieve a −30 dB Taylor weighted distribution. The simulation results show that the designed phased array antenna has a good one‐dimensional phased scanning ability of ±45° with active voltage standing wave ratio less than 2 at 75–79 GHz, along with −20 dB low sidelobe symmetrical radiation patterns in the H plane. An antenna sample fed by a WR12 waveguide is fabricated and measured to verify its performance. The measured VSWR is less than 2 at 73.6–85 GHz. This design provides a new idea for millimetre wave automotive radar antenna design.

Abstract Background Overweight and obesity have become major public health concerns worldwide. Persistent stress can activate the human hypothalamic‒pituitary‒adrenal axis (HPA) and increase the intake of “self-rewarding food”, thereby raising the incidence of obesity. Health care workers (HCWs) experience higher workloads and mental stress than workers in many other industries, which may put them at increased risk for overweight/obesity. However, few studies have been carried out on overweight and obesity among HCWs in China, and the overall scenario and behind-the-scenes factors of their overweight and obesity are unknown. The aim of this study is to understand the epidemic of overweight and obesity and risk factors among Chinese HCWs. Methods Based on a cross-sectional web survey design, 23,234 HCWs from 100 health institutions in 5 provinces/autonomous regions/municipalities across China were sampled to answer a self-administered questionnaire that was purposely developed using a multi-staged clustered random-sampling method. Chi-square test and ANOVA were performed to compare variables between two or more groups. Univariate analyses were conducted to identify the influence of self-reported persistent stress and/or recurrent anxiety/depressed mood on lifestyle behaviors. A multivariate binary logistic regression model was used to analyse the risk factors of overweight/obesity. Results Among the respondents, 34.26% were overweight, and 11.22% were obese. Most of the respondents had regular exercise habits (68.17%), had habitually stayed-up late (65.06%) and had been affected by persistent stress and/or recurrent anxiety/depressed mood (62.04%). A higher proportion of those with persistent stress and/or recurrent anxiety/depressed mood than those without habitually staying-up late (76.18%); consumed take-out food (54.92%), fried food (49.93%), snacks or desserts (50.51%); drank sugary drinks (46.57%); smoked (14.27%); and drank alcohol (23.34%). Gender (Female) (OR: 0.314, 95%CI: 0.292–0.336), age (OR: 1.742–2.334, 95%CI: 1.544–2.858), education (OR: 0.620–0.728, 95%CI: 0.445–0.973), living and working area (OR: 1.271, 95%CI: 1.192–1.355), breakfast (OR: 0.898, 95%CI: 0.839–0.960), fried food (OR: 1.133, 95%CI: 1.048–1.224), and alcohol consumption (OR: 1.111, 95%CI: 1.017–1.214) were factors for overweight/obesity. All of the aforementioned results were significant (P

The plant pathogen Botrytis cinerea (B. cinerea) causes severe plant diseases worldwide. Trichoderma is widely used as a biocontrol agent against B. cinerea through multiple biocontrol mechanisms. However, Trichoderma spp. with high biocontrol efficiency against B. cinerea under low-temperature conditions are barely reported. This study aimed to find potential low-temperature resistance biocontrol Trichoderma spp. against B. cinerea, and to characterize the biological principles underlying the activity of Trichoderma. Trichoderma koningiopsis (T. koningiopsis) C5-9 could fully overgrow a B. cinerea colony at 16 °C in a dual-culture assay. Treatment of cucumber leaves with T. koningiopsis C5-9 fermentation broth using the dipping method prior to B. cinerea inoculation significantly reduced the necrotic lesion diameter, with an inhibition rate of 55.30%. T. koningiopsis C5-9 could be successfully cultivated using the mycelia of B. cinerea as a carbon source at 16 °C. Transcriptomic analysis indicated that the origin recognition complex, organic substance catabolic process, and peroxisome were involved in the responses of T. koningiopsis C5-9 to B. cinerea. The findings of this study not only identified T. koningiopsis C5-9 as a potential biological control agent inhibiting B. cinerea under low-temperature conditions, but also provided new insights to develop a deeper understanding of the activity of Trichoderma against B. cinerea for plant protection.

Abstract Oat bran (OB) is a by‐product of oat, which is rich in β‐glucan. As a new food processing technology, ultrafine powder can improve the surface properties of samples. OB with different grinding times was prepared, and its functional components, physical properties, adsorption properties, and antioxidant properties were evaluated. Results showed that with increased grinding times, the average particle size of OB decreased significantly (p

The reported genome-walking methods still suffer from some deficiencies, such as cumbersome experimental steps, short target amplicon, or deep background. Here, a simple and practical fork PCR was proposed for genome-walking. The fork PCR employs a fork primer set of three random oligomers to implement walking task. In primary fork PCR, the low-stringency amplification cycle mediates the random binding of primary fork primer to some places on genome, producing a batch of single-stranded DNAs. In the subsequent high-stringency amplification, the target single-strand is processed into double-strand by the site-specific primer, but a non-target single-stranded DNA cannot be processed by any primer. As a result, only the target DNA can be exponentially amplified in the remaining high-stringency cycles. Secondary/tertiary nested fork PCR(s) further magnifies the amplification difference between the both DNAs by selectively enriching target DNA. The applicability of fork PCR was validated by walking several gene loci. The fork PCR could be a perspective substitution for the existing genome-walking schemes.

Suppression of transcription in S-phase is crucial to prevent genome instability. Zhang et al demonstrate that increase of H4K20me1 due to loss of Kmt5b cause genome instability in muscle stem cells, resulting in stem cell senescence but rhabdomyosarcoma formation when p53 is inactivated.

Abstract Background Brassinosteroids (BRs) are a group of essential steroid hormones involved in diverse developmental and physiological processes in plants. The Brassinazole-resistant 1 (BZR1) transcription factors are key components of BR signaling and integrate a wide range of internal and environmental signals to coordinate plant development, growth, and resistance to abiotic and biotic stresses. Although the BZR1 family has been fully studied in Arabidopsis, celery BZR1 family genes remain largely unknown. Results Nine BZR1 genes were identified in the celery genome, and categorized into four classes based on phylogenetic and gene structure analyses. All the BZR1 proteins shared a typical bHLH (basic helix-loop-helix) domain that is highly conserved across the whole family in Arabidopsis, grape, lettuce, ginseng, and three Apiaceae species. Both duplications and losses of the BZR1 gene family were detected during the shaping of the celery genome. Whole-genome duplication (WGD) or segmental duplication contributed 55.56% of the BZR1 genes expansion, and the γ as well as celery-ω polyploidization events made a considerable contribution to the production of the BZR1 paralogs in celery. Four AgBZR1 members (AgBZR1.1, AgBZR1.3, AgBZR1.5, and AgBZR1.9), which were localized both in the nucleus and cytoplasm, exhibit transcription activation activity in yeast. AgBZR1.5 overexpression transgenic plants in Arabidopsis showed curled leaves with bent, long petioles and constitutive BR-responsive phenotypes. Furthermore, the AgBZR1 genes possessed divergent expression patterns with some overlaps in roots, petioles, and leaves, suggesting an extensive involvement of AgBZR1s in the developmental processes in celery with both functional redundancy and divergence. Conclusions Our results not only demonstrated that AgBZR1 played a conserved role in BR signaling but also suggested that AgBZR1 might be extensively involved in plant developmental processes in celery. The findings lay the foundation for further study on the molecular mechanism of the AgBZR1s in regulating the agronomic traits and environmental adaptation of celery, and provide insights for future BR-related genetic breeding of celery and other Apiaceae crops.

Utrophin is a dystrophin-related protein stabilizing the sarcolemma in absence of dystrophin. Here the authors report that inactivation of the protein deacetylase SIRT6, involved in the deacetylation of the epigenetic mark H3K56ac in muscle cells, increases expression of utrophin and ameliorates dystrophic muscle pathology in mice.

Abstract Background Many factors can affect the recurrence of endometriosis after surgery, however, whether endometriotic cyst fluid contributes to endometriosis recurrence after surgical excision of ovarian endometriomas remains unclear. The objective of this study was to determine the presence of endometrial cells in ovarian endometriosis cyst fluid and the potential differences between these cells and those in the cyst wall. Methods Samples of cyst fluid (n = 39) and drainage fluid (n = 14) were collected from patients with ovarian endometriomas undergoing laparoscopic surgery. Drainage fluid from 14 patients without endometriosis was used as a control. The presence of endometrial cells in cyst fluid and drainage fluid was determined by cell culture in vitro and immunostaining. In addition, cyst fluid endometrial fragments and viscosity were analysed by transcriptome sequencing analysis and apparent diffusion coefficients, respectively. An animal model was used to confirm the ability of endometrial cells in cyst fluid to form new lesions. Results We found endometrium-like tissues in 71.8% (28/39) of cyst fluid and 71.4% (10/14) of drainage fluid samples by histopathological examination, and the presence of endometrioid tissue in cyst fluid was related to the viscosity of the cyst fluid. The living endometrial cells in cyst fluid and drainage fluid were confirmed by cell culture in vitro and immunostaining. Moreover, the adhesion ability of endometrial fragments in cyst fluid was significantly higher than that of ectopic tissues in the cyst wall (P

Abstract Mental fatigue increases risk-taking behavior. Using data collected between June 15 and August 6, 2020, this study investigates the impact of miners’ mental fatigue on risk decision-making to improve risk prevention and prediction abilities, and to reduce the occurrence of coal mine safety accidents. A total of 273 and 33 people participated in the preliminary and formal experiments, respectively. The participants, coal miners, visited a lab thrice to complete the pre-experiment, Balloon Analog Risk Task (BART), and Iowa Gambling Task (IGT). On the BART, mental fatigue displayed a significantly positive association with risk preference. On the IGT, as mental fatigue increased, net scores continuously decreased, while the frequency of making unfavorable decisions and the probability of taking risks increased. The BART value had no or weak correlations with the net score. Results suggest that mental fatigue leads to an increasing propensity to take risks. Therefore, regarding coal mine safety management, further attention is necessary concerning miners’ mental health, addressing mental fatigue, increasing rest time, and reducing night work. Furthermore, reasonable diet, improved working environments, and a positive attitude toward work should be promoted to reduce or eliminate mental fatigue and avoid decision-making errors that could cause accidents.

BackgroundHearing loss has occurred as a critical concern for aging and health. However, it remains unknown whether nocturnal sleep and midday napping duration are associated with hearing loss in middle-aged and older adults.MethodsThe study comprised 9,573 adults from China Health and Retirement Longitudinal Study, who have completed the survey for sleep characteristics and subjective functional hearing. We collected self-reported nocturnal sleep duration (30 min). The sleep information was classified into different sleep patterns. The primary outcome was self-reported hearing loss events. Multivariate Cox regression models and restricted cubic splines were used to investigate the longitudinal association of sleep characteristics with hearing loss. We applied Cox generalized additive models and bivariate exposure-response surface diagrams to visualize the effects of different sleep patterns on hearing loss.ResultsWe confirmed 1,073 cases of hearing loss (55.1% female) during the follow-up. After adjusting for demographic characteristics, lifestyle factors and health condition, nocturnal sleep with 9 h/night had higher risks of hearing loss.ConclusionInadequate nocturnal sleep was associated with an elevated risk of poor subjective hearing in middle-aged and older adults, while moderate napping decreased the risk of hearing loss. Keeping sleep stable within recommendation duration may be a useful strategy for preventing poor hearing loss.

Abstract Photoelectrochemical oxidation of methylene blue is investigated, with particular focus on the difference in kinetics and thermodynamics of decoloration and mineralization employing photoinduced absorption spectroscopy. Hematite and titania photoanodes are used for the comparison of both reactions, which is determined to be associated with the depth of the valence band (3.2 vs 2.5 V for titania and hematite, respectively). Methylene blue is mineralized by the titania photoanode, however it is only oxidized to small fragments by hematite. Such difference is related to the valence band potential that provides the thermodynamic driving force for photogenerated holes in both materials. In addition, the kinetic competition of water oxidation is found to occur on titania by controlling the pH of the electrolyte. In the pH 14 electrolyte, mineralization of methylene blue is suppressed due to the faster and dominant kinetics of water oxidation, in contrast to the complete mineralization in the near neutral electrolyte where water oxidation kinetics are modest. These results clearly address the importance considering both thermodynamic and kinetic challenges of methylene blue oxidation, which has been thought to be an easy molecule to oxidize, as the model reaction in the application of photo(electro)catalysis using metal oxides.

Pain is one of the main clinical symptoms of endometriosis, but its underlying mechanism is still not clear. Recent studies have shown that the secretory mediators of mast cells activated by estrogen are involved in the pathogenesis of endometriosis-related pain, but how estrogen-induced mast cell mediators are involved in endometriosis-related pain remains unclear. Here, mast cells were found to be increased in the ovarian endometriotic lesions of patients. They were also closely located closely to the nerve fibers in the ovarian endometriotic lesions from of patients with pain symptoms. Moreover, fibroblast growth factor 2 (FGF2)-positive mast cells were upregulated in endometriotic lesions. The concentration of FGF2 in ascites and the protein level of fibroblast growth factor receptor 1 (FGFR1) were higher in patients with endometriosis than in those without endometriosis, and they were correlated with pain symptoms. In vitro, estrogen could promote the secretion of FGF2 through G-protein-coupled estrogen receptor 30 (GPR30) via the MEK/ERK pathway in rodent mast cells. Estrogen-stimulated mast cells enhanced the concentration of FGF2 in endometriotic lesions and aggravated endometriosis-related pain in vivo. Targeted inhibition of the FGF2 receptor significantly restrained the neurite outgrowth and calcium influx in dorsal root ganglion (DRG) cells. Administration of FGFR1 inhibitor remarkably elevated the mechanical pain threshold (MPT) and prolonged the heat source latency (HSL) in a rat model of endometriosis. These results suggested that the up-regulated production of FGF2 by mast cells through non-classic estrogen receptor GPR30 plays a vital role in the pathogenesis of endometriosis-related pain.

Pesticides play a pivotal role in modern agriculture, but their potential environmental and health impacts necessitate a comprehensive understanding of their interactions with biological molecules. Beta-cyfluthrin, a widely used synthetic pyrethroid insecticide, is known for its efficiency in pest control. However, its interaction with bovine serum albumin (BSA), a crucial transport protein in living organisms, has not been extensively studied. The interaction between beta-cyfluthrin, a prominent synthetic pyrethroid insecticide, and bovine serum albumin (BSA) was comprehensively investigated using fluorescence spectrometry. Furthermore, the influence of ultraviolet (UV) degradation on the interaction parameters was explored, enhancing our understanding of the impact of environmental conditions on this interaction. The Stern–Volmer equation was employed to determine quenching constants, revealing that the fluorescence quenching mechanism primarily involved static quenching. The temperature variations were studied, showing an increase in the binding constant with rising temperature prior to degradation, while post-UV degradation, an inverse correlation between the binding constant and temperature was observed. The thermodynamic parameters were derived through appropriate equations, unveiling the underlying reaction forces. In the absence of degradation, hydrophobic interactions dominated, whereas after UV degradation, interactions shifted to hydrogen bonding and van der Waals forces. The findings elucidate the nuanced effects of UV degradation on the interaction between beta-cyfluthrin and BSA. This study furnishes critical insights that serve as a scientific foundation for pesticide production and application strategies, accounting for the influence of UV degradation on the intricate interplay between pesticides and BSA.

(E)-β-ocimene, a ubiquitous monoterpene volatile in plants, is emitted from flowers to attract pollinators and/or from vegetative tissues as part of inducible defenses mediated by complex signaling networks when plants are attacked by insect herbivores. Wild pear species Pyrus betuleafolia used worldwide as rootstock generally displays valuable pest-resistant traits and is a promising genetic resource for pear breeding. In the current study, transcriptional changes in this wild pear species infested with a polyphagous herbivore Spodoptera litura and the underlying molecular mechanisms were fully investigated. A total of 3,118 differentially expressed genes (DEGs) were identified in damaged pear leaf samples. Spodoptera litura larvae infestation activated complex phytohormonal signaling networks in which jasmonic acid, ethylene, brassinosteroids, cytokinin, gibberellic acid and auxin pathways were induced, whereas salicylic acid and abscisic acid pathways were suppressed. All DEGs associated with growth-related photosynthesis were significantly downregulated, whereas most DEGs involved in defense-related early signaling events, transcription factors, green leaf volatiles and volatile terpenes were significantly upregulated. The PbeOCS (GWHGAAYT028729), a putative (E)-β-ocimene synthase gene, was newly identified in P. betuleafolia transcriptome. The upregulation of PbeOCS in S. litura-infested pear leaves supports a potential role for PbeOCS in herbivore-induced plant defenses. In enzyme-catalyzed reaction, recombinant PbeOCS utilized only geranyl pyrophosphate but not neryl diphosphate, farnesyl pyrophosphate or geranylgeranyl diphosphate as a substrate, producing (E)-β-ocimene as the major product and a trace amount of (Z)-β-ocimene. Moreover, as a catalytic product of PbeOCS, (E)-β-ocimene showed repellent effects on larvae of S. litura in dual-choice bioassays. What is more, (E)-β-ocimene increased mortalities of larvae in no-choice bioassays. These findings provide an overview of transcriptomic changes in wild pears in response to chewing herbivores and insights into (E)-β-ocimene biosynthesis in pear plants, which will help elucidate the molecular mechanisms underlying pear-insect interactions.

Owing to its potential role in improving intestinal health, oat bran has received a lot of attention. In vitro digestion and fermentation were performed on oat brans that had been pretreated with steaming, microwave, and hot-air drying. The results showed that the dietary fiber after in vitro digestion and β-glucan content of pretreated oat brans changed insignificantly (p > 0.05). The abundance of Firmicutes and Proteobacteria varied significantly between samples (p

The limitations of the current genome-walking strategies include strong background and cumbersome experimental processes. Herein, we report a genome-walking method, fusion primer-driven racket PCR (FPR-PCR), for the reliable retrieval of unknown flanking DNA sequences. Four sequence-specific primers (SSP1, SSP2, SSP3, and SSP4) were sequentially selected from known DNA (5'→3′) to perform FPR-PCR. SSP3 is the fragment that mediates intra-strand annealing (FISA). The FISA fragment is attached to the 5′ end of SSP1, generating a fusion primer. FPR-PCR comprises two rounds of amplification reactions. The single-fusion primary FPR-PCR begins with the selective synthesis of the target first strand, then allows the primer to partially anneal to some place(s) on the unknown region of this strand, producing the target second strand. Afterward, a new first strand is synthesized using the second strand as the template. The 3′ end of this new first strand undergoes intra-strand annealing to the FISA site, followed by the formation of a racket-like DNA by a loop-back extension. This racket-like DNA is exponentially amplified in the secondary FPR-PCR performed using SSP2 and SSP4. We validated this FPR-PCR method by identifying the unknown flanks of Lactobacillus brevis CD0817 glutamic acid decarboxylase genes and the rice hygromycin gene.

Drought stress often occurred in citrus to limit its growth, distribution, and fruit quality. Cuticular waxes play an important role in regulating plant tolerance to drought stress. Plant enoyl-CoA reductase (ECR) is involved in the biosynthesis of cuticular waxes and catalyzes the last step of very long-chain fatty acids (VLCFAs) elongation. In this study, a putative ECR gene, named CsECR, was cloned from “Newhall” navel orange. CsECR protein has high identities with other plant ECR proteins and contained a conserved NADP/NAD-binding motif and three conserved functional sites. The highest expression of CsECR was observed in leaves, followed by stems, flavedos, ovaries, juice sacs, stigmas, stamens, albedos, and petals. Besides, the expression of CsECR was significantly induced by PEG6000 and ABA treatments. Ectopic overexpression of CsECR increased the contents of total waxes and aliphatic wax fractions (n-fatty acids, unsaturated fatty acids, n-alkanes, alkenes, iso-, and anteiso-alkanes) in the leaves and fruits of the transgenic tomato. Furthermore, ectopic overexpression of CsECR reduced the cuticle permeability in the leaves and fruits of the transgenic tomato and increased its tolerance to drought stress. Taken together, our results revealed that CsECR plays an important role in plant response to drought stresses by regulating cuticular wax biosynthesis.

Gamma-aminobutyric acid (GABA) has positive effects on many physiological processes. Lactic acid bacterial production of GABA is a future trend. This study aimed to produce a sodium-ion-free GABA fermentation process for Levilactobacillus brevis CD0817. In this fermentation, both the seed and fermentation media used L-glutamic acid instead of monosodium L-glutamate as the substrate. We optimized the key factors influencing GABA formation, adopting Erlenmeyer flask fermentation. The optimized values of the key factors of glucose, yeast extract, Tween 80, manganese ion, and fermentation temperature were 10 g/L, 35 g/L, 1.5 g/L, 0.2 mM, and 30 °C, respectively. Based on the optimized data, a sodium-ion-free GABA fermentation process was developed using a 10-L fermenter. During the fermentation, L-glutamic acid powder was continuously dissolved to supply substrate and to provide the acidic environment essential for GABA synthesis. The current bioprocess accumulated GABA at up to 331 ± 8.3 g/L after 48 h. The productivity of GABA was 6.9 g/L/h and the molar conversion rate of the substrate was 98.1%. These findings demonstrate that the proposed method is promising in the fermentative preparation of GABA by lactic acid bacteria.

Abstract Background Endometriosis is a refractory and recurrent disease and it affects nearly 10% of reproductive-aged women and 40% of infertile patients. The commonly accepted theory for endometriosis is retrograde menstruation where endometrial tissues invade into peritoneal cavity and fail to be cleared due to immune dysfunction. Therefore, the comprehensive understanding of immunologic microenvironment of peritoneal cavity deserves further investigation for the previous studies mainly focus on one or several immune cells. Results High-quality transcriptomes were from peritoneal fluid samples of patients with endometriosis and control, and firstly subjected to 10 × genomics single-cell RNA-sequencing. We acquired the single-cell transcriptomes of 10,280 cells from endometriosis sample and 7250 cells from control sample with an average of approximately 63,000 reads per cell. A comprehensive map of overall cells in peritoneal fluid was first exhibited. We unveiled the heterogeneity of immune cells and discovered new cell subtypes including T cell receptor positive (TCR+) macrophages, proliferating macrophages and natural killer dendritic cells in peritoneal fluid, which was further verified by double immunofluorescence staining and flow cytometry. Pseudo-time analysis showed that the response of macrophages to the menstrual debris might follow the certain differentiation trajectory after endometrial tissues invaded into the peritoneal cavity, that is, from antigen presentation to pro-inflammation, then to chemotaxis and phagocytosis. Our analyses also mirrored the dysfunctions of immune cells including decreased phagocytosis and cytotoxic activity and elevated pro-inflammatory and chemotactic effects in endometriosis. Conclusion TCR+ macrophages, proliferating macrophages and natural killer dendritic cells are firstly reported in human peritoneal fluid. Our results also revealed that immune dysfunction happens in peritoneal fluid of endometriosis, which may be responsible for the residues of invaded menstrual debris. It provided a large-scale and high-dimensional characterization of peritoneal microenvironment and offered a useful resource for future development of immunotherapy.

Skeletal muscle stem cells (MuSCs), also called satellite cells, are instrumental for postnatal muscle growth and skeletal muscle regeneration. Numerous signaling cascades regulate the fate of MuSCs during muscle regeneration but the molecular mechanism by which MuSCs sense mechanical stimuli remain unclear. Here, we describe that Piezo1, a mechanosensitive ion channel, keeps MuSCs in a quiescent state and prevents senescence. Absence of Piezo1 induces precocious activation of MuSCs, attenuates proliferation, and impairs differentiation, essentially abolishing efficient skeletal muscle regeneration and replenishment of the MuSC pool. Furthermore, we discovered that inactivation of Piezo1 results in compensatory up-regulation of T-type voltage-gated Ca2+ channels, leading to increased Ca2+ influx, which strongly induces NOX4 expression via cPKC. Elevated NOX4 expression in Piezo1-deficient MuSCs increases ROS levels and DNA damage, causing P53-dependent cellular senescence and cell death. The importance of the P53/P21-axis for mediating Piezo1-dependent cellular defects was confirmed by pharmacological inhibition of P53 in Piezo1-deficient mice, which abrogates increased senescence of muscle cells and normalizes muscle regeneration. Our findings uncover an essential role of Piezo1-mediated mechano-signaling in MuSCs for maintaining quiescence and preventing senescence. Reduced mechano-signaling due to decreased physical activity during aging may contribute to the increase of senescent cells and the decline of MuSC numbers in geriatric mice and humans.

Using oat-corn-konjac extruded mixed powder, oat bran micro powder, skim milk powder, Pueraria whole powder, and pumpkin powder as raw materials, a formula powder with high dietary fiber was prepared, and its effect on obesity in mice with a high-fat diet was investigated. After 7 days of adaptive feeding, the mice were divided into blank group, high-fat diet group, formula powder + high-fat diet group, and weight-loss drug + high-fat diet group. After 8 weeks of treatment, the body weight of mice were observed and measured to determine the composition of tract flora, liver leptin content, insulin content, and activities of AMP-activated protein kinase (AMPK), lipoprotein lipase (LPL), fatty acid synthetase (FAS), sterol-regulatory element-binding proteins (SREBPs), and acetyl CoA carboxylase 1 (ACC1). The results indicated that treatment with the formula powder could reduce the body weight of mice and increase the abundance of Bifidobacterium, Akkermansia, and Romboutsia compared to the group given a high-fat diet. Moreover, the leptin and insulin contents of the experimental group decreased from 5.67 μg/L to 0.12 μg/L and from 12.71 μg/L to 7.13 μg/L, respectively, compared to the control group, which was not significantly different from the blank group (P > 0.05). Also, the activities of AMPK and LPL increased, and the activities of FAS, SREBPs, and ACC1 were significantly decreased (P < 0.05). Some pathogenic bacteria were significantly positively correlated with leptin and FAS and significantly negatively correlated with LPL. Some beneficial bacteria were positively correlated with LPL. Therefore, the formula powder used in this study could reduce the body weight of mice, increase the abundance of some beneficial bacteria in the colonic intestinal microbiota, and improve the activities of enzymes related to lipid metabolism in the liver. This study provides a theoretical reference for the pathway by which high-fiber diet improves liver and intestinal metabolic abnormalities.

Hardware prefetching can seriously interfere with Flush+Reload cache side channel attack. This interference is not taken into consideration in previous Flush+Reload attacks. In this paper, an improved Flush+Reload is provided which minimizes the impact of hardware prefetchers. Specifically, prefetching is analyzed based on reverse engineering and the result is used to make an evaluation model to evaluate the impact of hardware prefetching on Flush+Reload attacks. Then the model is applied to fine tune the placement of probes in Flush+Reload attack to mitigate the prefetching impact. The experiments show that the approach is effective on the Core i5 processor which is equipped with highly aggressive prefetchers.

Although long non coding RNAs (lncRNAs) and circular RNAs (circRNAs) play important roles in the pathogenesis of diseases, endometriosis related lncRNAs and circRNAs are still rarely reported. This study focused on the potential molecular mechanism of endometriosis related competitive endogenous RNA (ceRNA) composed of lncRNAs and circRNAs. We performed high-throughout sequencing of six normal endometria, six eutopic endometria and six ectopic endometria for the first time to describe and analyze the expression profile of lncRNA, circRNA and mRNA. Our results showed that 140 lncRNAs, 107 circRNAs and 1,206 mRNAs were differentially expressed in the ectopic group, compared with the normal and eutopic groups. We established an lncRNA/circRNA-mRNA co-expression network using pearson correlation test. Meanwhile, the results of Gene set enrichment analysis analysis showed that the 569 up-regulated differentially expressed mRNA (DEmRNA) were mainly related to the epithelial-mesenchymal transition, regulation of immune system process and immune effector process. Subsequently, we established a DElncRNA-miRNA and DEcircRNA-miRNA network using the starbase database, identified the common miRNAs and constructed DElncRNA/DEcircRNA-miRNA pairs. miRDB, Targetscan, miRwalk and circRNA/lncRNA-mRNA pairs jointly determined the miRNA-mRNA portion of the circRNA/lncRNA-miRNA-mRNA co-expression network. RT-qPCR results of 15 control samples and 25 ectopic samples confirmed that circGLIS2, circFN1, LINC02381, IGFL2-AS1, CD84, LYPD1 and FAM163A were significantly overexpressed in ectopic tissues. In conclusion, this is the first study to illustrate ceRNA composed of differentially expressed circRNA, lncRNA and mRNA in endometriosis. We also found that lncRNA and circRNA exerted a pivotal function on the pathogenesis of endometriosis, which can provide new insights for further exploring the pathogenesis of endometriosis and identifying new targets.

Based on ultraviolet absorption spectroscopy technology combined with stoichiometry, a portable photoelectric detection system with wireless transmission was designed with the advantages of simple operation, low cost, and quick response to realize the non-destructive detection of dihydrocoumarin content in coconut juice. Through the detection of a sample solution, the light intensity through the solution is measured and converted into absorbance. Particle swarm optimization (PSO) is applied to optimize support vector regression (SVR) to establish a corresponding concentration prediction model. At the same time, in order to solve the shortcomings of the conventional portable photoelectric detection equipment in data storage, data transmission, and other aspects, based on the optimal PSO-SVR model, we used Python language to develop a friendly graphical user interface (GUI), integrating data collection, storage, analysis, and prediction modeling in one, greatly simplifying the operation process. The experimental results show that, compared with the traditional methods, the system achieves the purpose of rapid and non-destructive detection and has a small gap compared with the detection results of the ultraviolet spectrophotometer. It provides a good method for the determination of dihydrocoumarin in coconut juice.

Pretreatment improves the edible quality of oat bran and prolongs the shelf life, whereas the effect of pretreatments (i.e., steaming(S-OB), microwaving(M-OB), and hot-air drying(HA-OB)) on the flavor characteristics of oat bran is unknown. This study identified volatile composition using HS-SPME/GC–MS and an electronic nose of oat bran. The amino acid compositions were determined by a High-Speed automatic amino acid analyzer and the fatty acids were determined by gas chromatography. The results showed that steaming and microwaving pretreatments enhanced the nutty notes of oat bran. Sixty-four volatile compounds in four oat brans were identified. OB exhibited higher aroma-active compounds, followed by S-OB, and M-OB, and the HA-OB had the lowest aroma-active compounds. Hexanal, nonanal, (E)-2-octenal,1-octen-3-ol, 2-ethylhexan-1-ol, and 2-pentylfuran were the key volatile compositions in oat bran. The aldehyde content decreased and the esters and ketones increased in steamed oat bran. Microwaving and hot air drying increased the aldehyde content and decreased the ester and alcohol content. Steamed oat bran had the lowest levels of total amino acids (33.54 g/100 g) and bitter taste amino acids (5.66 g/100 g). However, steaming caused a significant reduction in saturated fatty acid content (18.56%) and an increase in unsaturated fatty acid content (79.60%) of oat bran (p < 0.05). Hot air drying did not result in an improvement in aroma. The results indicated that steaming was an effective drying method to improve the flavor quality of oat bran.

Endometriosis is an estrogen-dependent gynecological disease. The pathogenesis of endometriosis remains controversial, although it is generally accepted that the inflammatory immune response plays a crucial role in this process. Mast cells (MCs) are multifunctional innate immune cells that accumulate in endometriotic lesions. However, the molecular mechanism by which estrogen modulates MCs in the development of endometriosis is not well understood. Here we report that estrogen can induce the expression of NOD-like receptor family pyrin domain containing 3 (NLRP3) through estrogen receptor (ER)-α via the estrogen responsive element (ERE) in MCs. Such transcriptional regulation is necessary for the activation of NLRP3 inflammasome and the production of mature interleukin (IL)-1β in MCs. Targeted inhibition of NLRP3 significantly restrained lesion progression and fibrogenesis in a mouse model of endometriosis. Collectively, these findings suggest that MCs contribute to the development of endometriosis through NLRP3 inflammasome activation mediated by nuclear-initiated estrogen signaling pathway.

Background: SARS-CoV-2 spreads rapidly around the world, and some patients present gastrointestinal symptoms. The existence of the virus in the gastrointestinal tract makes digestive endoscopy a high-risk operation, which associated with an increased risk of infection rate in healthcare workers. This study aimed at exploring current knowledge, practice and attitudes of healthcare workers in endoscopy units in China regarding the status of occupational protection during COVID-19 pandemic.Methods: A cross-sectional study of a national online survey involving 717 healthcare workers in endoscopy units from 94 medical structures in 24 provinces and municipalities around China was conducted online via a questionnaire platform called Wenjuanxing (wjx.cn). The data were analyzed using correlation approaches, Kruskal-Wallis test for independent samples, and linear regression models.Results: Most Chinese healthcare workers in endoscopy units had a good knowledge of COVID-19 (median: 10; range: 7–12), showed a strikingly positive attitude (median: 65; range: 39–65), and carried out good practice (median: 47; range: 14–50) in strengthening the protection, disinfection and management of COVID-19. In terms of attitudes, female staff was more concerned about protection against COVID-19 than male staff (KW = 8.146, P = 0.004). Nurses performed better in both attitude (KW = 2.600, P = 0.009) and practice (KW = 6.358, P < 0.001) than endoscopic physicians when carrying out personal protection, patient care and environmental disinfection against SARS-CoV-2 infection. More positive attitudes in protection were related to better protective behavior in endoscopic daily medical work (r = 0.312; P < 0.001).Conclusion: The findings of this study suggest that Chinese endoscopy healthcare workers have an excellent mastery of knowledge about COVID-19, which is transformed into positive beliefs and attitudes, contributing to good practice during daily endoscopic procedures. Medical staff may benefit from further education. With the gradual normalization amid the ongoing COVID-19 pandemic, protection and management in endoscopy units may be changed accordingly.

The high integration of power information physical system improves the efficiency of power transmission, but it also brings new threats to power grid. False data injection attacks can use traditional bad data to detect vulnerabilities and maliciously tamper with measurement data to affect the state estimation results. In order to achieve a higher security level for power systems, we propose an earth mover distance method to detect false data injection attacks in smart grids. The proposed method is built on the dynamic correlation of measurement data between adjacent moments. Firstly, a joint-image-transformation-based scheme is proposed to preprocess the measurement data variation, so that the distribution characteristics of measurement data variation are more significant. Secondly, the deviation between the probability distribution of measurement data variation and the histogram are obtained based on the earth’s mover distance. Finally, a reasonable detection threshold is selected to judge whether there are false data injection attacks. The proposed method is tested using IEEE 14 bus system considering the state variable attacks on different nodes. The results verified that the proposed method has a high detection accuracy against false data injection attacks.

Background: The density and the activity of mast cells are associated with endometriosis. However, the role of mast cells on the pathogenesis of endometriosis remains unclear. Our study aims to investigate whether endometrial cells interact with mast cells and the involvement of their crosstalk in the development of endometriosis. Methods: The transwell assay was applied to investigate the effect of mast cells on the migratory ability of human primary endometrial cells. Mast cells were cocultured with endometrial epithelial and stromal cells respectively and total RNAs were isolated and subjected to mRNA sequencing. Next, the transwell assay, CCK-8, and tube formation were applied to study the role of CCL8 on the endometrial and endothelial cells in vitro. The mouse model was also established to confirm the role of CCL8 in the development and angiogenesis of endometriosis. Results: CCL8 was up-regulated in mast cells when cocultured with endometrial cells. CCL8 was highly expressed in the ectopic endometrium and the serum of patients with endometriosis. CCL8 promoted the migratory ability of endometrial epithelial and stromal cells and increased the proliferation, migration, and tube formation of endothelial cells. CCR1, the receptor of CCL8, was over-expressed in the ectopic endometrium and colocalized with blood vessels in ovarian endometriomas. The inhibition of CCR1 suppressed the development and angiogenesis of endometriosis in vivo. Conclusion: The crosstalk between endometrial cells and mast cells in the development of endometriosis via CCL8/CCR1 was demonstrated, thereby providing a new treatment strategy for endometriosis.

Employing mycorrhiza-defective mutants and their progenitors does not require inoculation or elimination of the resident microbial community in the experimental study of mycorrhizal soil ecology. We aimed to examine the suitability of mycorrhiza-defective rice (non-mycorrhizal, Oryza sativa L., cv. Nipponbare) and its progenitor (mycorrhizal) to evaluate nitrogen (N) loss control from paddy fields via arbuscular mycorrhizal (AM) fungi. We grew the two rice lines in soils with the full community of AM fungi and investigated root AM colonization. In the absence of AM fungi, we estimated rice N content, soil N concentration and microbial community on the basis of phospholipid fatty acids; we also quantified N loss via NH3 volatilization, N2O emission, runoff and leaching. In the presence of AM fungi, we did not find any evidence of AM colonization for non-mycorrhizal rice while mycorrhizal rice was colonized and percentage of root colonization was 17–24%. In the absence of AM fungi, the two rice lines had similar N content, soil N concentration and microbial community. Importantly, there was no significant difference in N loss via all the four pathways between mycorrhizal and non-mycorrhizal systems. This mycorrhizal/non-mycorrhizal rice pair is suitable for further research on the role of AM fungi in the control of soil N loss in paddy fields.

Farfarae flos (FF) is widely used for cough over thousands of years in China, but little is known about their pharmacokinetics properties. This study was aimed to establish a rapid and accurate ultraperformance liquid chromatography with triple-quadrupole tandem mass spectrometry method for compare pharmacokinetics studies of eight active compounds after oral administration between raw and honey-processed farfarae flos extracts. Optimum separation was performed on a Thermo Hypersil GOLD C18 column (100 mm × 2.1 mm, 1.9 µm particles size) with a gradient elution of acetonitrile as mobile phase A and 0.3% formic acid aqueous solution as mobile phase B. The flow rate was set as 0.3 mL/min and separated for 34.0 minutes. Electrospray ionization in the negative ion mode and selected reaction monitoring were used to identify and separate active components. The results met the acceptance criteria and showed that this method exhibited good linear, precision, accuracy, and stability. The extraction recoveries ranged from 81.54% to 104.48%, and the matrix effects ranged from 81.94% to 103.02%. These results show that the validated method could be successfully applied to evaluate the pharmacokinetic study in rats after oral administration of raw farfarae flos (R-FF) and honey-processed farfarae flos (H-FF).

Visible light communication (VLC) based on a light-emitting diode (LED) is considered to be a potential candidate for the next-generation communication. In this paper, a novel Zadoff–Chu matrix transform (ZCT) precoding is proposed to improve the performance of the traditional space-time block coding- (STBC-) based multiple-input multiple-output and orthogonal frequency division multiplexing (MIMO-OFDM) system. Compared with the existing orthogonal circulant matrix transform (OCT) precoding scheme, the proposed ZCT precoding achieves a much lower peak-to-average power ratio (PAPR) while maintaining the advantage of the uniform signal-to-noise ratio (SNR), which reduces the performance loss caused by LED nonlinearity. To study the system performance further, we set up an experimental demonstration to verify performance improvement under the condition of different driving peak-to-peak voltages (Vpps) and direct current (DC) offsets. Experimental results show that ZCT precoding gains the best bit error rate (BER) performance compared with the traditional and the OCT precoding MIMO-OFDM systems, whose BER is always below the 7% pre-forward error correction (pre-FEC) threshold of 3.8 × 10−3.

In this paper, an orthogonal circulant matrix transform (OCT) precoding technique is proposed to combine with the entropy loading in the multiple-input multiple-output and orthogonal frequency division multiplexing (MIMO-OFDM) visible light communication (VLC) system where the space-time coding (STBC) is chosen for its robustness to the channel correlation. Benefitting from the OCT precoding technique, the uniform signal-to-noise ratio (SNR) among all the subchannels can be achieved. As a result, only one SNR value is required to be fed back, and the same distribution matcher is employed during probabilistic shaping (PS), which means much lower feedback overhead and system complexity than the conventional entropy loading scheme. Experimental results show that the OCT precoding does not cause the system performance loss where the achievable information rate (AIR) of the proposed system is comparable with the conventional system without precoding. With an available bandwidth of ∼25 MHz, the proposed scheme can realize the AIR of 50.75 Mb/s at the expense of 0.45% average forward error correction (FEC) overhead (OH).

Farfarae Flos (FF) is the dried flower bud of Tussilago farfara L, which has antitussive, expectorant, and anti-inflammatory effects. However, little research on the main active composition of FF has been reported. The purpose of this study is to find the main active compounds responsible for the three pharmacological effects (i.e., antitussive, expectorant, and anti-inflammatory effects) of Farfarae Flos, based on the spectrum−effect relationship combined with chemometrics. First, this study uses the UPLC-QDA method to establish the chromatography fingerprint of Farfarae Flos, which is combined with chemometrics to analyze 18 batches of samples. Then, we study the antitussive, expectorant, and anti-inflammatory effects of Farfarae Flos. Finally, the spectrum−effect relationship between the fingerprint and the three pharmacological effects are studied by grey correlation analysis and partial least squares regression. The results show that four, four, and three main active constituents were found for the antitussive, expectorant, and anti-inflammatory pharmacological effects, respectively. In conclusion, we found the main active compounds corresponding to the main pharmacodynamic effects of Farfarae Flos. To our knowledge, this is the first time that spectrum−effect relationships in FF have been established using both raw and processed samples, which provides an experimental basis for further studies on the pharmacodynamic material basis of Farfarae Flos, as well as providing reference for the comprehensive evaluation of Farfarae Flos quality and the development of substitute resources.

Angiosperms, the flowering plants, provide the essential resources for human life, such as food, energy, oxygen, and materials. They also promoted the evolution of human, animals, and the planet earth. Despite the numerous advances in genome reports or sequencing technologies, no review covers all the released angiosperm genomes and the genome databases for data sharing. Based on the rapid advances and innovations in the database reconstruction in the last few years, here we provide a comprehensive review for three major types of angiosperm genome databases, including databases for a single species, for a specific angiosperm clade, and for multiple angiosperm species. The scope, tools, and data of each type of databases and their features are concisely discussed. The genome databases for a single species or a clade of species are especially popular for specific group of researchers, while a timely-updated comprehensive database is more powerful for address of major scientific mysteries at the genome scale. Considering the low coverage of flowering plants in any available database, we propose construction of a comprehensive database to facilitate large-scale comparative studies of angiosperm genomes and to promote the collaborative studies of important questions in plant biology.

With the rapid development of light-emitting diode, visible light communication (VLC) has become a candidate technology for the next generation of high-speed indoor wireless communication. In this paper, we investigate the performance of the 32-quadrature amplitude modulation (32-QAM) constellation shaping schemes for the first time, where two special circular constellations, named Circular (4, 11, 17) and Circular (1, 5, 11, 15), and a triangular constellation are proposed based on the Shannon’s criterion. Theoretical analysis indicates that the triangular constellation scheme has the largest minimum Euclidian distance while the Circular (4, 11, 17) scheme achieves the lowest peak-to-average power ratio (PAPR). Experimental results show that the bit error rate performance is finally decided by the value of PAPR in the VLC system due to the serious nonlinearity of the LED, where the Circular (4, 11, 17) scheme always performs best under the 7% preforward error correction threshold of 3.8 × 10−3 with 62.5Mb/s transmission data rate and 1-meter transmission distance.

Light-emitting diode- (LED-) based visible light communication (VLC) has become a potential candidate for next generation high-speed indoor wireless communication. Due to the limited modulation bandwidth of the LED, orthogonal frequency division multiplexing (OFDM) modulation is particularly preferred in the VLC system to overcome the ISI, which suffers from the high peak-to-average power ratio (PAPR) and leads to severe performance loss. In this paper, we propose and experimentally demonstrate a novel Zadoff-Chu matrix (ZCM) precoding scheme, which can not only reduce the PAPR, but also provide uniform signal-to-noise ratio (SNR) profile. The theoretical analysis and simulation show that the proposed scheme achieves better PAPR performance compared with the traditional precoding schemes. The experimental demonstration further validates the bit error rate (BER) performance improvement, where the measured BERs are all below the 7% pre-forward error correction (pre-FEC) limit of 3.8 × 10−3 when the transmitted data rate is 50 Mb/s.

In this paper, we propose and experimentally demonstrate a switching multiple input and multiple output (MIMO) system combining with adaptive orthogonal frequency division multiplexing (OFDM) modulation for high-speed indoor visible light communications. The adaptive OFDM modulation, which is realized by power and bit allocation on OFDM subchannels, is utilized to achieve the maximum channel capacity under a given target bit error rate (BER). Meanwhile, the MIMO mode switches between spatial multiplexing and transmit diversity adapting to the channel correlation, where the modulation order solved by adaptive OFDM modulation is chosen as the switching criterion. Experimental results validate data rates improvement over the pure spatial multiplexing and the pure transmit diversity system, where BERs are all below the 7% preforward error correction (pre-FEC) threshold of 3.8 × 10−3 in experiments.

In the study, a surface plasmon resonance-based (SPR-based) competitive assay was performed to analyze different compounds’ inhibitory activity to TNF-α, an important pro-inflammatory cytokine in the pathogenesis of chronic inflammatory diseases. Moreover, the single mass spectrometry (MS) detection method was coupled with an ultra-high-performance liquid chromatography (UPLC) system for the routine quality control (QC) of a traditional Chinese medicine (TCM). The above quality control strategy was evaluated with Lonicera japonica Thunb. Analytes were firstly separated on a Waters ACQUITYTM UPLC HSS T3 column (2.1 × 50 mm; particle size = 1.8 μm) using a 0.1% formic acid gradient elution, then detected by negative ESI mass spectrometry. The limits of quantification (LOQ) for analytes reached 0.005−0.56 μg/mL. The LOD of the QDa detector was lower than that of the PDA detector, indicating its wider detection range. The QDa detector was also more suitable for the analysis of the complex matrix of TCM. The method showed excellent linearity, with regression coefficients higher than 0.9991. The average recoveries of the investigated analytes were in the range of 98.78−105.13%, with an RSD below 3.91%. The inter-day precision range (n = 3 days) was 2.51−4.54%. Compared to other detectors, this strategy could be widely applied in the quantitative analysis of TCM. In addition, the chemically latent data could be revealed using chemometric analysis. Importantly, this study provides an efficient screening method for small-molecule inhibitors targeting the TNF-α pathway.

The purpose of this study was to establish a rapid, reliable, and sensitive ultra-performance liquid chromatography with triple-quadrupole tandem mass spectrometry coupled with chemometric method to measure and evaluate the differences between thirteen compounds in raw and processed Tussilago farfara L. from different sources. This assay method was validated, and the results indicated that the calibration curves for the thirteen compounds had good linearity (R2 > 0.9990). The limits of detection and limits of quantification of the thirteen compounds ranged from 0.0012 to 0.0095 μg/mL and from 0.0038 to 0.0316 μg/mL, respectively. The relative standard deviations (RSD) of the intra- and inter-day precisions and stability ranged from 1.06 to 2.00%, 0.26 to 1.99%, and 0.75 to 1.97%, respectively. The sample recovery rates of the thirteen compounds with different concentrations were 94.47⁻104.06%. The chemometric results, including principal component analysis, hierarchical clustering analysis, three-dimensional analysis, and box plot analysis, indicated that there are significance differences in raw and processed Tussilago farfara L. The results of this study confirm that the proposed method is the first reported method that has been successfully applied for simultaneous determination and discovery of the difference between thirteen compounds of raw and processed Tussilago farfara L. Thus, this method could be a helpful tool for the detection and confirmation of the quality of traditional Chinese medicines and provide a basis for future pharmacological studies.

An accurate and reliable method using ultra-high performance liquid chromatography combined with triple quadrupole tandem mass spectrometry (UHPLC–MS/MS) was established for simultaneous quantification of five major bioactive analytes in raw, wine-processed, and salt-processed Radix Achyranthis bidentatae (RAB). The results showed that this method exhibited desirable sensitivity, precision, stability, and repeatability. The overall intra-day and inter-day variations (RSD) were in the range of 1.57–2.46 and 1.51–3.00%, respectively. The overall recoveries were 98.58–101.48% with a relative standard deviation (RSD) of 0.01–1.86%. In addition, the developed approach was applied to 21 batches of raw, wine-processed, and salt-processed samples of RAB. Hierarchical clustering analysis (HCA), principal component analysis (PCA), heat map, and boxplot analysis were performed to evaluate the quality of raw, wine-processed, and salt-processed RAB collected from different regions. The chemometrics combined with the quantitative analysis based on UHPLC–MS/MS results indicated that the content of five analytes increased significantly in processed RAB compared to raw RAB.

As a valuable herbal medicine, the fruits of Xanthium strumarium L. (Xanthii Fructus) have been widely used in raw and processed forms to achieve different therapeutic effects in practice. In this study, a comprehensive strategy was proposed for evaluating the active components in 30 batches of raw and processed Xanthii Fructus (RXF and PXF) samples, based on high-performance liquid chromatography coupled with photodiode array detection (HPLC-PDA). Twelve common peaks were detected and eight compounds of caffeoylquinic acids were simultaneously quantified in RXF and PXF. All the analytes were detected with satisfactory linearity (R2 > 0.9991) over wide concentration ranges. Simultaneously, the chemically latent information was revealed by hierarchical cluster analysis (HCA) and principal component analysis (PCA). The results suggest that there were significant differences between RXF and PXF from different regions in terms of the content of eight caffeoylquinic acids. Potential chemical markers for XF were found during processing by chemometrics.