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Hypocotyl length is a botanical trait that affects the cold tolerance of Brassica napus L. (B. napus). In this study, we constructed an F2 segregating population using the cold-resistant short hypocotyl variety '16VHNTS158' and the cold-sensitive long hypocotyl variety 'Tianyou 2288' as the parents, and BSA-seq was employed to identify candidate genes for hypocotyl length in B. napus. The results of parental differences showed that the average hypocotyl lengths of '16VHNTS158' and 'Tianyou 2288' were 0.41 cm and 0.77 cm at the 5~6 leaf stage, respectively, after different low-temperature treatments, and '16VHNTS158' exhibited lower relative ion leakage rates compared to 'Tianyou 2288'. The contents of indole acetic acid (IAA), gibberellin (GA), and brassinosteroid (BR) in hypocotyls of '16VHNTS158' and 'Tianyou 2288' increased with decreasing temperatures, but the IAA and GA contents were significantly higher than those of 'Tianyou 2288', and the BR content was lower than that of 'Tianyou 2288'. The genetic analysis results indicate that the genetic model for hypocotyl length follows the 2MG-A model. By using SSR molecular markers, a QTL locus associated with hypocotyl length was identified on chromosome C04. The additive effect value of this locus was 0.025, and it accounted for 2.5% of the phenotypic variation. BSA-Seq further localized the major effect QTL locus on chromosome C04, associating it with 41 genomic regions. The total length of this region was 1.06 Mb. Within this region, a total of 20 non-synonymous mutation genes were identified between the parents, and 26 non-synonymous mutation genes were found within the pooled samples. In the reference genome of B. napus, this region was annotated with 24 candidate genes. These annotated genes are predominantly enriched in four pathways: DNA replication, nucleotide excision repair, plant hormone signal transduction, and mismatch repair. The findings of this study provide a theoretical basis for cloning genes related to hypocotyl length in winter rapeseed and their utilization in breeding. [ABSTRACT FROM AUTHOR]

There are more than 1000 species of microbes reside in the human gut, umbering∼1014 microbes. As the invisible organ of human beings, gut microbiota can usually participate in drug metabolism by producing specific enzymes, such as reductase and hydrolytic enzyme, thus affecting the efficacy, toxicity, and bioavailability of drugs. At least 30 commercially available drugs have been shown to be substrates of gut microbes-derived enzymes, and an increasing number of drugs may have the potential to contact with the distal gut with the help of improved release systems or poor solubility/permeability, more drugs are expected to be found to be metabolized through the gut flora. By collecting examples of intestinal flora participating in the metabolism of synthetic drugs and traditional Chinese medicine components, this article provides a comprehensive reference for future researchers to study drug metabolism by intestinal flora. Noticeably, the composition and quantity of intestinal flora varies among individuals, and can be affected by some drug administration (such as antibiotics) or environmental changes (acute plateau hypoxia). This seems to suggest that intestinal flora could have the potential to be a new drug target to affect the efficacy of drugs which can be metabolized by Intestinal flora. Accordingly, understanding the impact of intestinal flora on drug metabolism and clarifying the drug transformation process is of great significance for guiding rational clinical use, individualized use, toxicological evaluation, and promoting drug discovery and development. [ABSTRACT FROM AUTHOR]

1. Pinoresinol di-O-β-d-glucopyranoside (PDG), geniposide (GE), geniposidic acid (GA), aucubin (AN) and chlorogenic acid (CA) are the representative active ingredients inEucommiae cortex(EC), which may be estrogenic. 2. The ultra high-performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous determination of the five ingredients showed good linearity, low limits of quantification and high extraction recoveries, as well as acceptable precision, accuracy and stability in mice plasma and tissue samples (liver, spleen, kidney and uterus). It was successfully applied to the comparative study on pharmacokinetics and tissue distribution of PDG, GE, GA, AN and CA between normal and ovariectomized (OVX) mice. 3. The results indicated that except CA, the plasma and tissue concentrations of PDG, GE, GA in OVX mice were all greater than those in normal mice. AN could only be detected in the plasma and liver homogenate of normal mice, which was poorly absorbed in OVX mice and low in other measured tissues. PDG, GE and GA seem to be better absorbed in OVX mice than in normal mice proved by the remarkable increased value of AUC0–∞andCmax. It is beneficial that PDG, GE, GA have better plasma absorption and tissue distribution in pathological state. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Eucommiae cortex (EC), the bark of Eucommia ulmoides Oliv., has been traditionally used to treat many diseases in China for more than 2000 years. The pharmacological effects are primarily attributed to the presence of lignans, iridoids and phenolics, which are main active ingredients in EC. Aim of the study First, to investigate the active ingredients that can be absorbed into the rat plasma according to which ingredients exhibit significant correlation of drug concentration–time curve. Second, to establish an efficient ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of ingredients absorbed in rat plasma. Finally, to investigate gender effect on the pharmacokinetics of the ingredients absorbed in male and female rats plasma after oral administration with EC extract. Materials and methods 18 ingredients from EC were detected by UPLC-MS/MS, 9 out of 18 ingredients were absorbed into rat plasma. And 5 ingredients exhibit significant correlation of drug concentration–time curve. They were pinoresinol di-O- β -d-glucopyranoside (PDG), geniposide (GE), geniposidic acid (GA), aucubin (AN) and chlorogenic acid (CA). The analytes were extracted from rat plasma via a simple protein precipitation procedure and osalmid was used as the internal standard. Chromatographic separation was achieved on a Waters ACQUITY HSS T3 column (2.1 mm×100 mm, 1.8 μm) using a gradient elution program with acetonitrile and 0.1% formic acid water as the mobile phase, with a flow rate of 0.3 mL min −1 . The detection was performed on a triple-quadrupole tandem mass spectrometer by multiple reactions monitoring (MRM) mode in a positive ion mode via electrospray ionization (ESI). The transition monitored were / z 683.00[M+H] + →235.10 for PDG, / z 389.00[M+H] + →208.80 for GE, m / z 375.00[M+H] + →194.79 for GA, m / z 364.00[M+NH 4 ] + →148.81 for AN, m / z 355.10[M+H] + →162.84 for CA and m / z 230.03[M+H] + →120.77 for internal standard. Results The developed method showed good linearity over a wide concentration range, the lower limits of quantification and higher accuracy and precision for determination of the 5 analytes. Then the method was applied to study the pharmacokinetics in rats, and the results indicated that there were significant differences in pharmacokinetic parameters of the analytes between the male and female rats, and absorptions of these analytes in male group were all significantly higher than those in female group. Conclusion This study established an efficient, sensitive and selective UPLC-MS/MS method for simultaneous determination of the five ingredients in rat plasma, and it could be successfully applied to the comparative pharmacokinetic studies in male and female rats after oral administration with EC extract. [ABSTRACT FROM AUTHOR]

BACKGROUND: Biogenic amines (BA) are hazardous components in Huangjiu (HJ). To ensure the quality of Dangshen Huangjiu (DSHJ), an orthogonal experiment L9 (33) was proposed to optimize the process by the main brewing factors (pre‐fermentation temperature, pre‐ and post‐fermentation time) that may affect BA and functional factors in DSHJ. DSHJ was produced with low BA content and high functional factors. Gas chromatography–ion mobility spectrometry combined with a multivariate statistical method (GC‐IMS‐MSM) was used to analyze the volatile components in the brewing process of DSHJ. RESULTS: The optimum brewing process parameters of DSHJ were as follows: pre‐fermentation temperature, 28 °C; pre‐fermentation time, 9 days; post‐fermentation time, 18 days. The average content of BA in DSHJ was 33.12 mg L−1, and the sensory score, total phenol content and DPPH free radical scavenging rate of DSHJ were significantly higher than those of HJ. A total of 14 esters, 7 acids, 7 alcohols, 1 ketone, 5 aldehydes and 1 pyrazine in DSHJ and HJ were identified by GC‐IMS. There were no significant differences (P > 0.05) in DSHJ and HJ in the soaking rice and saccharification stage. 11 components, such as ethyl acetate, and 12 components, such as acetic acid, were the different components of HJ and DSHJ in pre‐fermentation and post‐fermentation stages, respectively. In the post‐fermentation stage, the contents of 8 components in DSHJ such as ethyl acetate were higher than in HJ. CONCLUSION: The preparation process parameters of DSHJ optimized by orthogonal experiments can ensure that DSHJ has the advantages of low BA content, high total phenol content and good antioxidant activity. Sensory score and GC‐IMS‐MSM analysis found that DSHJ prepared using the optimal process had the characteristics of good taste and rich aroma. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

A novel electrochemical immunosensor for detecting potential depression biomarker Apolipoprotein A4 (Apo-A4) was developed using a multi-signal amplification approach. Firstly, the sensor utilized a modified electrode material, NG-PEI-COF, combining bipyridine-functionalized covalent organic framework (COF) and polyethyleneimine-functionalized nitrogen-doped graphene (NG-PEI), providing high surface area and excellent electron transfer capability for the first-stage amplification in electrical signal conduction. Subsequently, gold nanoparticles (AuNPs) were further electrodeposited onto the electrode, providing good biocompatibility and abundant binding sites for immobilizing the target antigen, thus achieving the second-stage amplification in target recognition and binding. To address the lack of redox properties of the antigen, a tracer probe was formed by loading AuNPs, anti-Apo-A4, and toluidine blue (TB) successively onto COF, leading to the third-stage amplification in signal conversion. The constructed electrochemical immunosensor TB/Ab/AuNPs/COF-Apo-A4/AuNPs/NG-PEI-COF/GCE exhibited excellent detection performance against Apo-A4 with a linear range of 0.01 to 300 ng mL−1 and had a low detection limit of 2.16 pg mL−1 (S/N = 3). In addition, the biosensor had good reproducibility (RSD = 2.31%), stability, and significant anti-interference performance toward other depression biomarkers. The sensor has been successfully used for the quantitative detection of Apo-A4 in serum, providing potential applications for detecting Apo-A4 in the clinic and serving as a reference for constructing sensing methods based on COF. [ABSTRACT FROM AUTHOR]

Owing to the increasing market demand of Codonopsis Radix, the cropper blindly cultivates to expand planting area for economic benefits, which seriously affects the quality of Codonopsis Radix. Therefore, this study synthesized 207 batches of Codonopsis Radix and 115 ecological factors, and analyzed the suitable planting areas of Codonopsis pilosula under current and future climate change based on Geographic Information System (GIS) and MaxEnt model. Secondly, we evaluated the quality of Codonopsis Radix based on the all-in-one functional factor including chromatographic fingerprint, the index components, the effective compounds groups, the nutritional components, and the nutritional elements, and the quality regionalization of Codonopsis Radix was analyzed. Finally, the ecological factors affecting the accumulation of effective components of Codonopsis Radix were analyzed. This study found for the first time that the highly suitable area of Codonopsis pilosula was mainly distributed in the Weihe River system and the Bailongjiang River system in Gansu Province. There were differences in the quality of Codonopsis Radix from different ecologically suitable areas based on the all-in-one functional factors, and the comprehensive high-quality area of Codonopsis Radix was mainly distributed in Longnan and Longxi district of Gansu Province. The precipitation, temperature and altitude play a key role in the accumulation of chemical components in the 10 ecological factors affecting the distribution of Codonopsis pilosula. Under future climatic conditions, the highly suitable area of Codonopsis pilosula is decreased. [ABSTRACT FROM AUTHOR]

A novel electrochemical immunosensor was developed for highly sensitive detection of brain-derived neurotrophic factor (BDNF), a well-known depression marker. The immunosensor was fabricated by modifying indium tin oxide-coated polyethylene terephthalate (ITO-PET) with N-doped graphene-polyaniline (NG-PANI) and gold nanoparticles (AuNPs) to enhance the conductivity and protein loading capacity. Subsequently, BDNF was immobilized onto the electrode surface via gold-sulfur bonds, followed by the attachment of biotinylated antibody (Biotin-Ab) and horseradish peroxidase-avidin (HRP-Avidin) to create the final immunosensor (HRP-Avidin–Biotin-Ab-BDNF-AuNPs/NG-PANI/ITO-PET). The proposed immunosensor exhibited a linear range of determination (0.781–400 pg/mL) with a low limit of detection (LOD) of 0.261 pg/mL (S/N = 3) and excellent reproducibility (RSD = 1.4%) and stability (92.7%, RSD = 3.1%). Additionally, the immunosensor demonstrated good anti-interference performance and good recovery (98.1–107%). To evaluate the practical utility of the immunosensor, BDNF levels were quantified in the serum of mice with depression induced by chronic unpredictable mild stress (CUMS). The results indicated that the serum BDNF levels were significantly decreased in the depression model group compared with the control group, highlighting the potential of this immunosensor for clinical detection of BDNF in depression diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Recently, we confirmed that the 95% ethanol‐extracted fraction of Codonopsis Radix, which contains several triterpenoids and sterols, possesses pharmacological activities. However, due to the low content and diverse types of triterpenoids and sterols, their similar structures, lack of ultraviolet absorption, and difficulty in obtaining controls, few studies have so far assessed their contents in Codonopsis Radix. We accordingly constructed an ultra‐high‐performance liquid chromatography‐quadrupole‐time‐of‐flight mass spectrometry technique for the simultaneous quantitative determination of 14 terpenoids and sterols. Separation was performed on the Waters Acquity UPLC HSS T3 C18 column (100 × 2.1 mm, 1.8 μm) with 0.1% formic acid (A) and 0.1% formic acid in methanol (B) as mobile phase under gradient elution. The determination coefficients for each of the matrix calibration curves were ≥0.9925. The average recovery ranged from 81.25% to 118.05%, with relative standard deviations of <4%. The contents of 14 components in 23 batches were quantified and further analyzed through chemometrics. Linear discriminant analysis can distinguish sample varieties. The quantitative analysis method can accurately determine the contents of 14 components and thereby provide the chemical basis for the quality control of Codonopsis Radix. It also could be a valuable approach for the classification of different Codonopsis Radix varieties. [ABSTRACT FROM AUTHOR]

Multi-elemental analysis is widely used to identify the geographical origins of plants. The purpose of this study was to explore the feasibility of combining chemometrics with multi-element analysis for classification of Codonopsis Radix from different producing regions of Gansu province (China). A total of 117 Codonopsis Radix samples from 7 counties of Gansu province were collected. Inductively coupled plasma mass spectrometry (ICP-MS) was used for the determination of 28 elements (39 K, 24 Mg, 44Ca, 27Al, 137Ba, 57Fe, 23Na, 88Sr, 55Mn, 66Zn, 65Cu, 85Rb, 61Ni, 53Cr, 51 V, 7Li, 208Pb, 59Co, 75As, 133Cs, 71 Ga, 77Se, 205Tl, 114Cd, 238U, 107Ag, 4Be and 202Hg). Among macro elements, 39 K showed the highest level, whereas 23Na was found to have the lowest content value. Micro elements showed the concentrations order of: 88Sr > 55Mn > 66Zn > 85Rb > 65Cu. Among trace elements, 53Cr and 61Ni showed higher content and 4Be was not detected in all samples. Intra-regions differentiation was performed by principal component analysis (PCA), cluster analysis (CA) and supervised learning algorithms such as linear discriminant analysis (LDA), k-nearest neighbors (k-NN), support vector machines (SVM), and random forests (RF). Among them, the RF model performed the best with an accuracy rate of 78.79%. Multi-elemental analysis combined with RF was a reliable method to identify the origins of Codonopsis Radix in Gansu province. [ABSTRACT FROM AUTHOR]

Cortisol, a corticosteroid hormone as a primary stress hormone response to internal and external stress, has been regarded as a gold standard reliable biomarker to evaluate human mental stress. The double enzymes strategy, using nanozyme and enzyme amplifying the electrochemical signal, has been widely used to improve the performance of electrochemical biosensors. An ultra-sensitive electrochemical cortisol sensor based on Au single-atom nanozymes had been fabricated through HRP labeled anti-cortisol antibody binding with Au by Au-S bond. Based on the high catalytic activity of Au single-atom nanozymes and the high selectivity of HRP-labeled anti-cortisol antibodies, the cortisol electrochemical sensor-based Au single-atom nanozymes had an excellent response to cortisol, such as high electrochemical activity, high sensitivity, high selectivity, and wide linear range (0.15–300 ng mL−1) and low detection (0.48 pg mL−1) through the four-parameter logistic model with 95% confidence. The electrochemical cortisol sensor was used to determine the cortisol concentration of human saliva at different times. The process of electrochemical biosensor fabrication. [Display omitted] • Zr MOF nanomaterials based on porphyrin. • Au-single atom nanozymes based on Zr porphyrin MOF. • Au single-atom nanozymes had high catalytic activity and high surface area. • A dual enzyme amplification strategy was used to fabricate an electrochemical cortisol sensor. • The electrochemical performance of the cortisol sensor had been amplified by Au single atom nanozymes and HRP labeled anit-cortisol. [ABSTRACT FROM AUTHOR]

Multi-elemental analysis is widely used to identify the geographical origins of plants. The purpose of this study was to explore the feasibility of combining chemometrics with multi-element analysis for classification of Codonopsis Radix from different producing regions of Gansu province (China). A total of 117 Codonopsis Radix samples from 7 counties of Gansu province were collected. Inductively coupled plasma mass spectrometry (ICP-MS) was used for the determination of 28 elements (39 K, 24 Mg, 44Ca, 27Al, 137Ba, 57Fe, 23Na, 88Sr, 55Mn, 66Zn, 65Cu, 85Rb, 61Ni, 53Cr, 51 V, 7Li, 208Pb, 59Co, 75As, 133Cs, 71 Ga, 77Se, 205Tl, 114Cd, 238U, 107Ag, 4Be and 202Hg). Among macro elements, 39 K showed the highest level, whereas 23Na was found to have the lowest content value. Micro elements showed the concentrations order of: 88Sr > 55Mn > 66Zn > 85Rb > 65Cu. Among trace elements, 53Cr and 61Ni showed higher content and 4Be was not detected in all samples. Intra-regions differentiation was performed by principal component analysis (PCA), cluster analysis (CA) and supervised learning algorithms such as linear discriminant analysis (LDA), k-nearest neighbors (k-NN), support vector machines (SVM), and random forests (RF). Among them, the RF model performed the best with an accuracy rate of 78.79%. Multi-elemental analysis combined with RF was a reliable method to identify the origins of Codonopsis Radix in Gansu province. [ABSTRACT FROM AUTHOR]

In recent years, the safety of Codonopsis Radix (CR) has attracted considerable attention. Pesticide residues is an important index to evaluate the safety of CR. The purpose of this study was to monitor pesticide residues in 164 batches of CR in China and assess dietary risk assessment. Firstly, a combined method of QuEChERS-GC–MS/MS and QuEChERS-LC–MS/MS was established for determination of 155 pesticide residues in CR. Second, 155 Pesticide residues in 3 CR cultivars from Gansu, Shanxi, Hubei, Guizhou and Chongqing were determined by this method. Finally, the risk score of pesticide residues in CR was evaluated, and the dietary health risk was evaluated based on the pesticide residues in CR. The results demonstrated that one or more pesticide residues were detected in 39 batches (23.78%) of 164 batches of CR. Of the 155 pesticide residues, 20 were detected. The most frequently detected pesticide residue was dimethomorph with a detection rate of 5.49%. Risk scores showed that 6 pesticides were at higher risk. Risk assessment based on the hazard quotient/hazard index (HQ/HI) approach revealed that exposure to pesticide residues which detected in CR were far below levels that might pose a health risk. [ABSTRACT FROM AUTHOR]

Connective tissue growth factor (CTGF) is a disease marker of rheumatoid arthritis (RA), and its rapid and sensitive detection is essential for the diagnosis of RA. In this work, a three-dimensional pore structure of alkali-activated nitrogen-doped graphene (aN-G) was used as an electrode modification material, and a label-free electrochemical immunosensor for the sensitive detection of CTGF was successfully constructed by the formation of an amide bond between amino groups in protein and carboxyl groups on the carbon surface. Under optimized conditions, the sensor achieved accurate detection of CTGF in the wide range of 0.0625 ~ 2000 pg mL−1. It had good accuracy (95.0 ~ 100.1%), repeatability (1.2 ~ 2.2%), stability, selectivity, and a low limit of detection (0.0424 pg mL−1, S/N = 3). The sensor was used in serum samples of patients with RA, and CTGF was also successfully detected. Based on this, the electrochemical sensor is expected to become an effective method for RA diagnosis and treatment effect evaluation. [ABSTRACT FROM AUTHOR]

The accurate and sensitive analysis of rheumatoid arthritis(RA) trace biomarkers is essential for early warning and diagnosis of RA, while anti‐cyclic citrullinated peptide antibody (anti‐CCP‐ab) is a specific one. In this study, a simple label‐free electrochemical immunosensor for the detection of anti‐CCP‐ab was constructed with nitrogen‐doped graphene (N−G) and gold nanoparticles (AuNPs). The proposed non‐enzyme immunosensor had exhibited high specificity, selectivity, and stability in the linear calibration curve range from 0.125∼2000 pg mL‐1 and has been successfully used in the detection of anti‐CCP‐ab in human serum, providing a new idea for the early diagnosis of RA. [ABSTRACT FROM AUTHOR]

Scheme The construction process of flexible immunosensor for DCNP1. [Display omitted] • A selective quantitative detection method for depression marker DCNP1 was established for the first time. • The construction of the label-free flexible immunosensor is facile, and the required detection time is short. • Pioneering application of NG/TiO 2 in the construction of immunosensors. • The developed immunosensor features a satisfactory linear range, low detection limit, excellent reproducibility, selectivity, and stability. • The constructed immunosensor has been successfully used to determine DCNP1 in brain tissue homogenate samples. In this work, a label-free flexible immunosensor for selective and sensitive detection of dendritic cell nuclear protein-1 (DCNP1), a newly discovered potential marker of depression, was successfully constructed. At first, a highly conductive nanomaterial nitrogen-doped graphene with titanium dioxide (NG/TiO 2) was prepared using a one-pot method, and it was used as an electrode modification material in the construction of immunosensors for the first time. NG/TiO 2 was functionalized with 1-pyrene-butyric acid N-hydroxysuccinimide ester (PSE), and then the DCNP1 antibody could be better immobilized on the surface of the modified indium tin oxide-polyethylene terephthalate (ITO-PET). Then bovine serum albumin (BSA) was used to block non-specific active sites, and the immunosensor (BSA/anti-DCNP1-PSE/NG/TiO 2 /ITO-PET) can be obtained. The fabricated NG/TiO 2 and the construction process of the immunosensor were characterized by scanning electron microscopy, infrared spectroscopy, cyclic voltammetry, and electrochemical impedance methods. Differential pulse voltammetry (DPV) was used to measure different concentrations of DCNP1 after optimizing the ratio of NG and TiO 2 , the modification amount of NG/TiO 2 , the ratio of NG/TiO 2 to PSE, the concentration of anti-DCNP1 and its immobilization time, the concentration of BSA, and the incubation time of DCNP1. The constructed sensor had a wide linear range of 1 ∼ 104 pg mL−1, and a low detection limit of 0.45 pg mL−1 (S/N = 3). It had good reproducibility (RSD = 1.90 %), repeatability (RSD = 2.6 %), and selectivity. After storing it at 4 ℃ for 15 days, its performance decreased to 92.37 %. And it had been successfully used to determine DCNP1 in the brain homogenate of mice with depression induced by chronic unpredictable mild stress (CUMS), providing a possible method for the rapid and accurate determination of DCNP1. [ABSTRACT FROM AUTHOR]

Winter rapeseed (Brassica rapa L.) is a major oilseed crop in Northern China, where its production was severely affected by chilling and freezing stress. However, not much is known about the role of differentially accumulated proteins (DAPs) during the chilling and freezing stress. In this study, isobaric tag for relative and absolute quantification (iTRAQ) technology was performed to identify DAPs under freezing stress. To explore the molecular mechanisms of cold stress tolerance at the cellular and protein levels, the morphological and physiological differences in the shoot apical meristem (SAM) of two winter rapeseed varieties, Longyou 7 (cold-tolerant) and Lenox (cold-sensitive), were explored in field-grown plants. Compared to Lenox, Longyou 7 had a lower SAM height and higher collar diameter. The level of malondialdehyde (MDA) and indole-3-acetic acid (IAA) content was also decreased. Simultaneously, the soluble sugars (SS) content, superoxide dismutase (SOD) activity, peroxidase (POD) activity, soluble protein (SP) content, and collar diameter were increased in Longyou 7 as compared to Lenox. A total of 6330 proteins were identified. Among this, 98, 107, 183 and 111 DAPs were expressed in L7 CK/Le CK, L7 d/Le d, Le d/Le CK and L7 d/L7 CK, respectively. Quantitative real-time PCR (RT-qPCR) analysis of the coding genes for seventeen randomly selected DAPs was performed for validation. These DAPs were identified based on gene ontology enrichment analysis, which revealed that glutathione transferase activity, carbohydrate-binding, glutathione binding, metabolic process, and IAA response were closely associated with the cold stress response. In addition, some cold-induced proteins, such as glutathione S-transferase phi 2(GSTF2), might play an essential role during cold acclimation in the SAM of Brassica rapa. The present study provides valuable information on the involvement of DAPs during cold stress responses in Brassica rapa L, and hence could be used for breeding experiments. [ABSTRACT FROM AUTHOR]

Background: Echinacoside (ECH) is the main active ingredient of Cistanches Herba, which is known to have therapeutic effects on metastatic tumors. However, the effects of ECH on liver cancer are still unclear. This study was to investigate the effects of ECH on the aggression of liver cancer cells. Methods: Two types of liver cancer cells Huh7 and HepG2 were treated with different doses of ECH at different times and gradients. MTT and colony formation assays were used to determine the effects of ECH on the viability of Huh7 and HepG2 cells. Transwell assays and flow cytometry assays were used to detect the effects of ECH treatment on the invasion, migration, apoptosis and cell cycle of Huh7 and HepG2 cells. Western blot analysis was used to detect the effects of ECH on the expression levels of TGF-β1, smad3, smad7, apoptosis-related proteins (Caspase-3, Caspase-8), and Cyto C in liver cancer cells. The relationship between miR-503-3p and TGF-β1 was detected using bioinformatics analysis and Luciferase reporter assay. Results: The results showed that ECH inhibited the proliferation, invasion and migration of Huh7 and HepG2 cells in a dose- and time-dependent manner. Moreover, we found that ECH caused Huh7 and HepG2 cell apoptosis by blocking cells in S phase. Furthermore, the expression of miR-503-3p was found to be reduced in liver tumor tissues, but ECH treatment increased the expression of miR-503-3p in Huh7 and HepG2 cells. In addition, we found that TGF-β1 was identified as a potential target of miR-503-3p. ECH promoted the activation of the TGF-β1/Smad signaling pathway and increased the expression levels of Bax/Bcl-2. Moreover, ECH could trigger the release of mitochondrial Cyto C, and cause the reaction Caspases grade. Conclusions: This study demonstrates that ECH exerts anti-tumor activity via the miR-503-3p/TGF-β1/Smad aixs in liver cancer, and provides a safe and effective anti-tumor agent for liver cancer. [ABSTRACT FROM AUTHOR]

Oligosaccharides are one of the major bioactive components in Codonopsis pilosula. The present study was to investigate the protective effect of C. pilosula oligosaccharides (CPO) on d-galactose (D-Gal)-induced aging and to illuminate the underlying mechanisms. The results indicated that CPO effectively increased the growth rate as indicated by body weight, attenuated the decline of thymus and liver indexes, and mitigated the pathological liver injury compared with model group rats. CPO could elevate the activities of anti-oxidative enzymes, including CAT, GSH-Px, and SOD, and reduce MDA level in serum, improve SOD and GSH-Px activities in liver, down-regulate the phosphorylation of MAPK cascade signal transduction components – ERK, JNK, and p38, and inhibit the activation of downstream NF-κB signaling. In addition, CPO treatment noticeably prevented the overexpression of inflammatory factors such as TNF-α, IL-6, and IL-1β in serum, inhibited the expression of apoptosis-related proteins caspase-3, caspase-9, and the ratio of Bax/Bcl-2 in liver caused by the inflammatory cascade, reduced cell apoptosis, and improved liver damage. The above studies suggested that CPO could effectively mitigate the D-Gal-induced aging, and the underlying mechanism might be closely related to ROS-induced oxidative stress, inflammation, and apoptosis. [ABSTRACT FROM AUTHOR]

Trace detection of argininosuccinate synthetase 1 (ASS1), a depression marker, in urine samples is difficult to achieve. In this work, a dual-epitope-peptides imprinted sensor for ASS1 detection in urine was constructed based on the high selectivity and sensitivity of the "epitope imprinting approach". First, two cysteine-modified epitope-peptides were immobilized onto gold nanoparticles (AuNPs) deposited on a flexible electrode (ITO-PET) by gold-sulfur bonds (Au–S), then a controlled electropolymerization of dopamine was carried out to imprint the epitope peptides. After removing epitope-peptides, the dual-epitope-peptides imprinted sensor (MIP/AuNPs/ITO-PET) which with multiple binding sites for ASS1 was obtained. Compared with single epitope-peptide, dual-epitope-peptides imprinted sensor had higher sensitivity, which presented a linear range from 0.15 to 6000 pg ml−1 with a low limit of detection (LOD = 0.106 pg mL−1, S/N = 3). It had good reproducibility (RSD = 1.74%), repeatability (RSD = 3.60%), stability (RSD = 2.98%), and good selectivity, and the sensor had good recovery (92.4%–99.0%) in urine samples. This is the first highly sensitive and selective electrochemical assay for the depression marker ASS1 in urine, which is expected to provide help for the non-invasive and objective diagnosis of depression. [Display omitted] • A dual-epitope imprinted sensor was constructed based on two different cysteine-modified peptides of ASS1. • This dual-epitope imprinted sensor enabled accurate determination of ASS1 in urine. • The sensitivity of dual-epitope (LOD = 0.106 pg/mL) was improved 2-fold compared to single-epitope (LOD = 0.232 pg/mL). • This dual-epitope imprinted sensor is expected to be used for objective diagnosis of depression. [ABSTRACT FROM AUTHOR]

Leptin is a vital biomarker of non-alcoholic fatty liver (NAFLD), and its evaluation of the concentration level in vivo is of great significance to NAFLD diagnosis. Therefore, it is pressing to develop a method for rapid and sensitive detection of leptin. This paper describes an environmentally friendly and label-free immunosensor based on porous graphene functionalized black phosphorus (PG-BP) composite to detect of leptin. The PG-BP was synthesized via strong coherent coupling between porous graphene (PG) surface plasmons and anisotropic black phosphorus (BP) localized surface plasmons, which made the electrochemical performance of PG and BP synergistic as well as increased the stability and conductive capability of BP material. The PG-BP modified electrodes was further prepared by gold nanoparticles, cysteamine, and glutaraldehyde in turn. Due to the cross-linking effect of glutaraldehyde, anti-leptin can be firmly fixed. These properties of the platform improved the conductive capability of the immunosensor and enhanced the load capacity of the proteins, thereby, the sensitivity of the immunosensor was significantly increased. Under the optimal conditions, the proposed immunosensor exhibited a wide linear range of 0.150–2500 pg/mL with a low detection limit of 0.036 pg/mL. The leptin immunosensor displayed excellent selectivity and anti-interference ability, which could be used for early screening and diagnosis of clinical NALFD. • A BSA/anti-leptin/Glu/Cys/AuNPs/PG-BP/GCE immunosensor was successfully fabricated. • The proposed immunosensor was successfully used for the detection of leptin with a wide range of detection and low detection limits. • The immunosensor can be used for the sensitive, efficient and real-time analysis of leptin in real samples. • The immunosensor is expected to provide a reliable, real-time and convenient method for clinical trials of NAFLD markers. [ABSTRACT FROM AUTHOR]

Abstract The porous graphene functionalized black phosphorus composite (PG-BP) was successfully synthesized via strong coherent coupling between porous graphene surface plasmons and anisotropic black phosphorus localized surface plasmons by the method of penetration of infrared ray using infrared drying. And it was used to fabricate a novel high-sensitive electrochemical sensor for determination of bisphenol A (BPA) in daily use and biological samples. The PG-BP had been characterized by infrared spectroscopy, Raman, X-ray photoelectron spectroscopic, scanning electron microscopy and energy dispersive spectrometer, indicating that PG-BP with the large specific surface area and good electrical conductivity was successfully prepared. In addition, electrochemical properties of the modified materials are comparably explored by means of impedance spectroscopy and cyclic voltammograms. Under the optimal conditions, BPA was determined quantitatively using differential pulse voltammetry. The results showed that the BPA detection current has a good linear relationship in the range of 4.3 × 10−8–5.5 × 10−5 mol/L with a detection limit of 7.8 × 10−9 mol/L (S/N = 3). Results from the present work illustrate the prominent performance of PG-BP as a sensing channel for BPA sensor application and the huge potential of PG-BP as an electrode material. Graphical abstract Schematic representation of PG-BP/GCE fabrication and electrochemical detection of BPA. Unlabelled Image Highlights • The PG-BP were successfully prepared based on the strong coupling between porous graphene and black phosphorus. • This is the first research on the electrochemical sensors based on PG -BP composite. • The proposed electrochemical sensor exhibited a broad detection range and low detection limit for BPA. • The electrochemical sensor can be used for the sensitive and efficient analysis of BPA in real samples. [ABSTRACT FROM AUTHOR]

Abstract Two water-soluble polysaccharides named CPP1a and CPP1c were isolated from C. pilosula Nannf. var. modesta L.T.Shen by hot-water extraction and purified by graded alcohol precipitation and DEAE-52 cellulose column. The structure of CPP1c with higher yield has been characterized while its antitumor activities has not been elucidated. In this study, we firstly analyzed the chemical structure of CPP1a. The results of instrumental analysis combined with chemical analysis showed that CPP1a was composed of →1)- β‑ l ‑Rha p ‑(4→, →1)- β‑Ara p ‑(5→, →1)- β‑ d ‑Gal p A‑(4→, →1)- β‑ d ‑Gal p ‑(6→, terminal‑β‑ d ‑Glc p in a molar ratio of 1:12:1:10:3 and its relative and absolute molecular weight were 1.01 × 105 Da and 1.03 × 105 Da respectively. Further, the cytotoxicity assay indicated that CPP1a and CPP1c were more sensitive to HepG2 cells than cervical carcinoma Hela cells and gastric carcinoma MKN45 cells. Both of CPP1a and CPP1c could influence cell morphology, inhibit the migration and induce apoptosis by affecting the G2/M phase of HepG2 cells. Preliminary mechanism studies confirmed that CPP1a and CPP1c could induce apoptosis through up-regulating the ratio of Bax/Bcl-2 and activating caspase-3. According to previous research, we might speculate that the reason for the stronger cytotoxicity and pro-apoptotic effect of CPP1c than that of CPP1a can be attributed to its high uronic acid content. [ABSTRACT FROM AUTHOR]

In this study, a novel electrochemical immunosensor for early screening of depression markers-heat shock protein 70 (HSP70) was successfully developed based on the porous graphene (PG) with huge specific surface area and excellent structure. Benefiting from the strong adsorption and good bioactivity of PG which was initially prepared via a simple pyrolysis process, a variety of heat shock protein70 (HSP70) can be firmly loaded on the PG to construct the basic electrode (HSP70/PG/GCE)，which was characterized by the cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), respectively. Due to the HSP70 fixed on the surface of basic electrode and the HSP70 in the samples can competitively combine with the horseradish peroxidase labeled human HSP 70 antibody (HRP-Strept-Biotin-Ab). As a result, it presented a negative correlation between the concentration of HSP70 in samples and the detection signal of the proposed electrochemical immunosensor (HRP-Strept-Biotin-Ab-HSP70/PG/GCE) in the test liquid. The application of PG with excellent electrical conductivity in construction of immunosensor remarkably improved the sensitivity of the immunosensor for detection of HSP70. The proposed immunosensor demonstrated a wide linear range of 0.0448 ~ 100 ng/mL with a low detection limit of 0.02 ng/mL at 3σ. Moreover, the proposed immunosensor could be applied for the sensitive and efficient detection of HSP70 in real samples with good precision, acceptable stability, reproducibility and satisfactory results. Therefore, the HSP70 immunosensor provides a novel and convenient method for early clinical screening of depression markers-heat shock protein 70. [ABSTRACT FROM AUTHOR]

In this study, polyaniline functionalized graphene quantum dots composite with excellent performance was successfully prepared by situ polymerized, and it was applied to fabricate electrochemical sensor for analysis of calycosin in the complex samples of traditional Chinese medicine. The polyaniline functionalized graphene quantum dots composite was characterized by Fourier transform infrared spectroscopy and Raman analysis, to indicate that polyaniline functionalized graphene quantum dots composite was successfully synthesized. The morphology and microstructure of the modified electrodes were characterized by Scanning electron microscope and Transmission electron microscope. According to the results of cyclic voltammetry and electrochemical impedance spectroscope, the glassy carbon electrode modified with polyaniline functionalized graphene quantum dots composite significantly increased the electron transfer rate on the surface of sensor. Under the optimized conditions, the fabricated sensor was used for the analysis of calycosin in Radix Hedysari and Radix Astragali by differential pulse voltammetry, which displayed acceptable reproducibility, long-term stability, good selectivity and sensitivity. The proposed sensor exhibited a wide detection range (1.1 × 10 −5 –3.52 × 10 −4  mol/L) with a detection limit of 9.8 × 10 −6  mol/L (S/N = 3). This study provided a novel and convenient method for rapid evaluation of the quality of Radix Hedysari and Radix Astragali . [ABSTRACT FROM AUTHOR]

A pectic polysaccharide (named as CPP1c) extracted from Codonopsis pilosula was evaluated for its structural features and potential of immune-modulating activities in an aging mouse model of senescence accelerated mouse prone 8 (SAMP8) in vitro and in vivo . The relative molecular weight and the absolute molecular weight of CPP1c were 1.26 × 10 5 Da and 1.49 × 10 5 Da, respectively. Investigation of structural features by a combination of chemical and instrumental analysis showed CPP1c was composed of →1)-α- l - R ha p -(2,4→, →1)-α- l -Ara f -(5→, →1)-α- d -Gal p -(6→ and →1)-α- d -Gal p A-(4→ in a molar ratio of 3:1:2:33. CPP1c could promote lymphocyte proliferation, modulate the percentage of CD4 + , CD8 + , CD28 + and CD152 + T cells and enhance the production of IL-2, TNF-α and IFN-γ. Moreover, PCR assay revealed CPP1c augmented the expressions of CD28, PI3 K and p38MAPK mRNA, and the increase of protein expressions of the same genes was also confirmed by western blot analyses. In addition, CPP1c had the potential of promoting the homing of lymphocytes. Taking all factors into consideration, we deduced CPP1c might exert its immunostimulating potency via promoting T cell activation by TCR/CD28 signaling pathways. [ABSTRACT FROM AUTHOR]

Fufang Ejiao Syrup (FES) is a syrup made from Colla Corii Asini (CCA) and four botanicals (Codonopsis Radix (CR), Ginseng Radix et Rhizoma Rubra (GRRR), Rehmanniae Radix Praeparata (RRP) and Crataegi Fructus (CF)) as a result of modern processing and refining technology. FES has a lengthy history and is frequently used in clinical practice. Modern pharmacological studies have confirmed that oligosaccharides in any of the main medicinal herbs of FES, such as CR, GRRR, and RRP, have significant immune-enhancing effects. Therefore, the oligosaccharide component in FES could be its important pharmacologic substance, however, no studies on the content, structural analysis and source attribution of oligosaccharides in FES have been reported. The objective of this study is to systematically analyze the oligosaccharide in FES, compare the differences of the major oligosaccharides in different batches of FES produced by one manufacturer, and construct the content determination method for determining the content of the major oligosaccharides in FES, to provide technical support for FES quality assessment. This analysis revealed that a total of 13 oligosaccharides were identified from the FES, including 3 disaccharides, 4 trisaccharides, 3 tetrasaccharides, and 3 pentasaccharides. The constructed UPLC-QQQ-MS fingerprint of FES oligosaccharide is simple, stable, and reproducible, making it a useful tool for assessing FES's quality. There was a significant difference between the oligosaccharide fingerprints of 16 batches of FES,the results of fingerprint analysis combined with the statistical analysis suggested that the differences in stachyose, sucrose and raffinose contents in FES may be the reason for the great variations in oligosaccharide fingerprints of different batches of FES. For the 5 oligosaccharides in FES, the UPLC-QQQ-MS technique showed significant linearity in the linear range, along with good stability, repeatability, and recovery. Mannotriose was found to be higher in FES, followed by sucrose and stachyose, while kestose and raffinose were relatively lower. The results of this study reveal that oligosaccharides are important components of FES, and the method of fingerprinting and content determination constructed has strong practical value and is expected to be used for FES quality control. • A systematic qualitative and quantitative study on FES. • 13 oligosaccharides were found from FES. • The oligosaccharide fingerprints of different batches of FES are different. • The content of 5 oligosaccharides with clear structure and origin was determined. [ABSTRACT FROM AUTHOR]

A novel lobetyolin electrochemical sensor based on a magnetic functionalized reduced graphene oxide/Nafion nanohybrid film has been introduced in this work. The magnetic functionalized reduced graphene oxide was characterized by fourier transform infrared spectroscopy, atomic force microscope, X-ray diffraction, transmission electron microscopy and thermogravimetric analysis. The scanning electron microscopy characterized the morphology and microstructure of the prepared sensors, and the electrochemical effective surface areas of the prepared sensors were also calculated by chronocoulometry method. The electrochemical behavior of lobetyolin on the magnetic functionalized reduced graphene oxide/Nafion nanohybrid modified glassy carbon electrode was investigated by cyclic voltammetry and differential pulse voltammetry in a phosphate buffer solution of pH 6.0. The electron-transfer coefficient (α), electron transfer number (n), and electrode reaction rate constant (Κs) were calculated as 0.78, 0.73, and 4.63 s − 1 , respectively. Under the optimized conditions, the sensor based on magnetic functionalized reduced graphene oxide/Nafion showed a linear voltammetric response to the lobetyolin concentration at 1.0 × 10 − 7 to 1.0 × 10 − 4 mol/L with detection limit (S/N = 3)of 4.3 × 10 − 8 mol/L. The proposed sensor also displayed acceptable reproducibility, long-term stability, and high selectivity, and performs well for analysis of lobetyolin in real samples. The voltammetric sensor was successfully applied to detect lobetyolin in Codonopsis pilosula with recovery values in the range of 96.12% –102.66%. [ABSTRACT FROM AUTHOR]

A sensitively competitive electrochemical immunosensor for the detection of cortisol was successfully developed based on gold nanoparticles and magnetic functionalized reduced graphene oxide (AuNPs/MrGO). In order to construct the base of the immunosensor, the MrGO was initially fabricated by chemical cross-linking and used to modify the nafion pretreated glassy carbon electrode. Subsequently, the surface of electrode was modified by AuNPs via electrochemical deposition. A variety of cortisol (Cor) can be firmly loaded in the AuNPs/MrGO with large specific surface area and good bioactivity to construct the basic electrode (Cor/AuNPs/MrGO/Nafion@GCE), which was characterized by the cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), respectively. Due to the cortisol on the surface of basic electrode and samples can competitively combine with the cortisol antibody labelled by horseradish peroxidase (HRP-Strept-Biotin-Ab). Finally, the detection signal of electrochemical immunosensor (HRP-Strept-Biotin-Ab-Cor/AuNPs/MrGO/Nafion@GCE) in the test liquid had negative correlations with the concentration of cortisol in samples. The AuNPs/MrGO with excellent electrical conductivity being applied, the electrochemical response of the immunosensor was immensely amplified. The immunosensor displayed excellent analytical performance for the detection of cortisol range from 0.1 to 1000 ng/mL with a detection limit of 0.05 ng/mL at 3σ. Moreover, compared the developed immunoassay with commercially available enzyme linked immunosorbent assay, the proposed method showed good precision, acceptable stability and reproducibility, indicating the immunosensor could be used for the sensitive, efficient and real-time detection of cortisol in real samples. Therefore, the present strategy provides a novel and convenient method for clinical determination of cortisol. [ABSTRACT FROM AUTHOR]

• Codonopsis Radix contains levan- and inulin-types of fructooligosaccharides. • Discrimination of 3 varieties of Codonopsis Radix using HPLC-ELSD fingerprints. • The LDA method can successfully distinguish 3 varieties of Codonopsis Radix. Codonopsis Radix (CR) is a plant that is important in the practice of traditional Chinese medicine (TCM). The Chinese Pharmacopoeia 2020 records dried roots prepared from three varieties of Campanulaceae plants under the designation CR ("Dang-shen" in Chinese), including Codonopsis pilosula (Franch.) Nannf (C. pilosula), Codonopsis pilosula Nannf.var. modesta (Nannf.) L81. T. Shen (C. pilosula var. modesta) and Codonopsis tangshen Oliv (C. tangshen). As major constituents of CR, oligosaccharides might contribute to its clinical efficacy except for other known active compounds, yet the differences in the oligosaccharide profiles of these three varieties of CR remain incompletely understood. In the present study, 135 samples from these different CR varieties were harvested, and oligosaccharide fingerprints for these samples were characterized via HPLC-ELSD, with 19 common peaks being matched. Oligosaccharides were further identified through the combination of electrospray ionization MS/MS (ESI-MS/MS) with nuclear magnetic resonance (NMR). Principal component analysis and linear discriminant analysis (LDA) approaches were then used to compare the oligosaccharide profiles of these three CR varieties. These analyses ultimately revealed that CR was compared with principal component analysis (PCA) and linear discriminant (LDA) methods. The analyses ultimately revealed that these CR samples contained high levels of inulin- and levan-type fructooligosaccharides (FOS), with variations in the relative levels of these FOS compounds among the three analyzed CR varieties. Through the combined analysis of oligosaccharide fingerprints and LDA results, it was possible to differentiate among these CR varieties, with an accurate classification rate of 96.3% and a cross-validation rate of 95.6%. Together, these results highlight a valuable approach to the classification and identification of different CR varieties. [ABSTRACT FROM AUTHOR]

Developing wearable flexible biosensors with excellent electrical conductivity and outstanding flexibility simultaneously for disease diagnosis and health monitoring is a current hot topic in the field of sensor research. In this study, nitrogen-doped graphene (N-Gr) and gold nanoparticles (AuNPs) were successively deposited onto ITO-PET flexible conductive films by chemical deposition, and then a flexible electrochemical immunosensor with high sensitivity and selectivity was constructed for detecting depression markers by exploiting the high affinity between AuNPs and the sulfhydryl groups of depression marker (DM) antibody. Due to the composite of N-Gr, AuNPs and ITO-PET, the prepared flexible sensor can maintain a relatively stable electrical signal response regardless of the deformation such as spiral, rolling, and bending, without shedding or fracture of the N-Gr and AuNPs, and can also significantly amplify the electrochemical response signal of the immunosensor. Under optimized experimental conditions, the fabricated immunosensor showed good linearity over a wide range of 0.023–300.00 ng/mL with a low detection limit of 0.010 ng/mL (3σ, n = 5), when it was used for the determination of depression markers-human Apo-A4 in 100% whole serum samples. The flexibility of the constructed immunosensor and the stability and sensitivity of biomolecular analysis are expected to be further made into implantable depression marker in-situ monitoring probe with the help of micro-machining technology, which will develop a promising method for objective, efficient and accurate clinical diagnosis of depression. Schematic representation of constructing AuNPs@N-Gr@ITO-PET-based electrochemical immunosensor for determination of apolipoprotein A4 (Apo-A4). [Display omitted] • A highly sensitive and selective flexible immunosensor for determination of human apolipoprotein A4 (Apo-A4) has been fabricated. • The prepared AuNPs@N-Gr with large specific surface area, excellent electrical conductivity and outstanding flexibility can amplify the signal of immunosensor. • The proposed immunosensor can be used to sensitively and efficiently analyze apolipoprotein A4 (Apo-A4) in serum samples. • This method would provide a promising potential in early clinical screening and diagnosis of depression. [ABSTRACT FROM AUTHOR]

Formononetin is an isoflavone compound found in many traditional Chinese medicines that has a wide range of pharmacological activities. It is critical to develop a sensitive, accurate, and rapid determination method for in-depth formononetin research. A molecularly imprinted electrochemical sensor was constructed in this study. After the flexible electrode (ITO-PET) was modified with nitrogen-doped graphene (NG), the molecularly imprinted polymer (MIP) was electropolymerized on the surface using o-phenylenediamine (OPD) as the functional monomer and formononetin as the template. Under optimized experimental conditions, the MIP sensor detected formononetin selectively in 0.1 M phosphate buffer solution (PBS) with the linear range of 3 ~ 120 μM, a detection limit as low as 1.14 μM (S/N = 3), and good anti-interference ability and reproducibility. The analytical performance of the proposed MIP/NG/ITO-PET was evaluated for the detection of formononetin in real samples such as methanol extract of Radix Astragali and mouse plasma with good accuracy and precision. [Display omitted] • A molecularly imprinted electrochemical sensor was established for the selective determination of formononetin for the first time. • The MIP is electropolymerized using OPD as the functional monomer and formononetin as the template. • The constructed MIP/NG/ITO-PET was successfully applied in complex systems such as astragalus extract and mouse plasma. • MIP/NG/ITO-PET has a broad linear range of 3 ~ 120 μM, with a low detection limit of 1.14 μM and good selectivity. [ABSTRACT FROM AUTHOR]

A sensitive electrochemical sensor based on bovine serum albumin (BSA)/poly (diallyldimethylammonium chloride) (PDDA) functionalized graphene nanosheets (PDDA-G) composite film modified glassy carbon electrode (BSA/PDDA-G/GCE) had been developed to investigate the oxidative protein damage and protections of protein from damage by flavonoids. The performance of this sensor was remarkably improved due to excellent electrical conductivity, strong adsorptive ability, and large effective surface area of PDDA-G. The BSA/PDDA-G/GCE displayed the greatest degree of BSA oxidation damage at 40 min incubation time and in the pH 5.0 Fenton reagent system (12.5 mM FeSO 4 , 50 mM H 2 O 2 ). The antioxidant activities of four flavonoids had been compared by fabricated sensor based on the relative peak current ratio of SWV, because flavonoids prevented BSA damage caused by Fenton reagent and affected the BSA signal in a solution containing Co(bpy) 3 3 + . The sensor was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM). UV–vis spectrophotometry and FTIR were also used to investigate the generation of hydroxyl radical and BSA damage, respectively. On the basis of results from electrochemical methods, the order of the antioxidant activities of flavonoids is as follows: (+)-catechin > kaempferol > apigenin > naringenin. A novel, direct SWV analytical method for detection of BSA damage and assessment of the antioxidant activities of four flavonoids was developed and this electrochemical method provided a simple, inexpensive and rapid detection of BSA damage and evaluation of the antioxidant activities of samples. [ABSTRACT FROM AUTHOR]

sCPP1b is a selenized derivative of pectic polysaccharide (CPP1b) that is isolated from Wen-Codonopsis pilosula. In this study, A549 cells were used for in vitro studies on cytotoxicity, cell morphology, cell migration, cycle distribution, apoptosis, and expression of apoptosis-associated proteins after treatment with CPP1b and sCPP1b. Results show that sCPP1b and CPP1b could change cell morphology, suppress cell migration, induce cell apoptosis, and block the cell cycle. CPP1b and sCPP1b induced apoptosis on A549 cells through upregulation of caspase-3 and Bax and downregulation of Bcl-2 proteins. sCPP1b possessed stronger antitumor activity than CPP1b. [ABSTRACT FROM PUBLISHER]

As a folk medicine, Aconitum sinomontanum Nakai (Ranunculaceae) a perennial herbaceous flowering plant, is a widely used traditional Chinese medicine. Its rhizomes and roots are known as ' Gaowutou ' in China, and it has been traditionally used for the treatment of rheumatoid arthritis, painful swelling of joints, bruises and injuries and has been known to grow well in regions of high altitude such as Gansu, Tibet etc. This systematic review the comprehensive knowledge of the A. sinomontanum , including its traditional processing and uses, chemical constituents, pharmacological activities, toxicity assessment, pharmacokinetics and metabolism, and its use in clinical settings to emphasize the benefits of this species. We also discuss expectations for prospective research and implementation of this herb. This work lays a solid foundation for further development of A. sinomontanum. Information on the studies of A. sinomontanum was collected from scientific journals, books, and reports via library and electronic data search (PubMed, Elsevier, Scopus, Google Scholar, Springer, Science Direct, Wiley, ACS, EMBASE, Web of Science and CNKI). Meanwhile, it was also obtained from published works of material medica, folk records, ethnopharmacological literatures, Ph.D. and Masters dissertation. As a member of the Ranunculaceae family, A. sinomontanum possesses its up-and-coming biological characteristics. It is widely reported for treating rheumatoid arthritis, painful swelling of joints, bruises and injuries. Currently, over 71 phytochemical ingredients have been obtained and identified from different parts of A. sinomontanum. Among them, alkaloids, flavonoids, steroids, glycosides are the major bioactive constituents. Activities such as antinociceptive, anti-inflammatory, antitumor, antiarrhythmic, local anesthetic, antipyretic, antimicrobial, insecticidal and others have been corroborated in vivo and in vitro. These properties are attributed to different alkaloids. In addition, many of the active ingredients, such as lappaconitine, ranaconitine and total alkaloids have been used as quality markers. This work contributes to update the ethnopharmacological uses, chemical constituents, pharmacological activities, toxicity assessment, pharmacokinetics and metabolism, and clinical settings information for A. sinomontanum , which provide basic information to help better understand the pharmacological and toxicological activities of A. sinomontanum in human. However, further in-depth studies are needed to determine the medical uses of this herb and its chemical constituents, pharmacological activities, clinical applications and toxicology. [Display omitted] [ABSTRACT FROM AUTHOR]

CPP1b is a pectic polysaccharide isolated from Codonopsis pilosula , and it possesses potent antitumor activity. In this study, a HNO 3 –Na 2 SeO 3 method was performed to synthesize selenium–CPP1b (sCPP1b). The effects of BaCl 2 usage amount and HNO 3 concentration on the yield and selenium content of sCPP1b were investigated by single-factor experiments. Reaction time, reaction temperature and the ratio of polysaccharide to Na 2 SeO 3 were optimized by orthogonal experimental design at three levels of each of the three factors (L 9 (3) 4 ) based on antitumor activity, selenium content and yield of sCPP1b. Our results showed that 5 h of reaction time, 60 °C of reaction temperature, and 2:2 ratio of polysaccharide to Na 2 SeO 3 were the optimal selenylation modification conditions. The validation experiments completed under the optimal conditions gave the mean selenium content and yield of sCPP1b were 478.17 μg/g (RSD = 5.7%) and 595 mg/g (RSD = 1.6%), respectively. Selenylation modification can significantly increase the antitumor activity of CPP1b in vitro . The structural characterization of sCPP1b was further characterized by Fourier-transform infrared spectroscopy, thermogravimetric analysis, and high-performance gel permeation chromatography coupled with multi-angle laser light scattering. These structure analysis results indicated that sCPP1b has been successfully selenylation modified with similar structure to polysaccharide of CPP1b. [ABSTRACT FROM AUTHOR]

The Funding section "Funding was funded by National Natural Science Foundation of China (Grants NO. 31960435, NO. 31860383), Research Program Sponsored by State Key Laboratory of Aridland Crop Science (Gansu Agricultural University), GSCS-2020-06, China Agriculture Research System of MOF and MARA, CARS-12, Research Program Sponsored by State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, No. GSCS-2020-Z1." now reads: "Funding was financially supported Research Program Sponsored by State Key Laboratory of Aridland Crop Science, Gansu Agricultural University (No. GSCS-2020-Z1); by the China Agriculture Research System of MOF and MARA (CARS-12); by the National Natural Science Foundation of China (NO. 31960435); by the Research Program Sponsored by State Key Laboratory of Aridland Crop Science (Gansu Agricultural University) (GSCS-2020-06) and the National Natural Science Foundation of China (NO. 31860388)." Correction to: I Scientific Reports i https://doi.org/10.1038/s41598-021-02707-z, published online 06 December 2021 The original version of this Article contained an error in the Funding section. Author Correction: iTRAQ-based quantitative proteome analysis insights into cold stress of Winter Rapeseed (Brassica rapa L.) grown in the field. [Extracted from the article]

Highlights: [•] A pectic polysaccharide (CPP1b) from Codonopsis pilosula was at first identified. [•] The chemical structure of CPP1b was elucidated. [•] The ultramicrostructure of CPP1b was described. [•] CPP1b presents potential anti-tumor and adjucant anti-tumor activity. [ABSTRACT FROM AUTHOR]

Abstract: A new sulfated acetamido-heteropolysaccharide, HPS4-2A, was obtained by aqueous extraction followed by precipitation with ethanol and fractionation with DEAE column chromatography from Radix Hedysari. It was composed of rhamnose, arabinose, glucose, galactose and 2-acetamido-2-deoxy-d-galactose in the molar ratio of 10.09%:25.90%:25.90%:25.0%:12.30%. Elemental analysis indicated that HPS4-2A was a sulfated polysaccharide containing small amount of sulfate groups (1.87%). Partial acid hydrolysis, GC, GC–MS, 1D and 2D NMR spectroscopy analysis of the HPS4-2A revealed a predominance of glucose, galactose and 2-acetamido-2-deoxy-d-galactose linked in a highly-branched structure. The molecular weight of HPS4-2A was determined by HPSEC and HPSEC-MALLS. AFM study indicated that HPS4-2A took a highly branched conformation, which in consistent with the result studied by SEC-MALLS. Structural features of HPS4-2A were also investigated by SEM and TEM. Antioxidant assays demonstrated that HPS4-2A possessed of strong DPPH and hydroxyl radicals scavenging activities, suggesting that HPS4-2A could potentially be used as natural antioxidant. [Copyright &y& Elsevier]

Abstract: In this work, the multi-walled carbon nanotubes-COOH (MWCNTs-COOH)/graphene (GNs)/gold nanoparticles (AuNPs) composite materials were synthesized by in situ growth of AuNPs on the surface of the MWCNTs-COOH/GNs. And then, the Bilirubin oxidase (BUD) was immobilized on the surface of the film to construct a biosensor for determination of Bilirubin. Morphology of the nanomaterials was analyzed by field-emission scanning electron microscopy, high resolution transmission electron microscopy, Brunauer–Emmett–Teller, and X-ray photoelectron spectroscopy. The performance of the biosensor was studied by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and amperometric response (i–t). It was demonstrated that the nanocomposite materials exhibited enhanced fast electron transfer and excellent biocompatibility. In this assay, the MWCNTs-COOH/GNs/AuNPs composite film based biosensor showed prominently electrocatalytic activity for the detection of Bilirubin with a linear range from 1.33μM to 71.56μM and the low limit of detection (LOD) is 0.34μM based on a signal to noise ratio (S/N=3). Meanwhile, the apparent Michaelis–Menten constant (K m ) was 64.86μM. In addition, this biosensor exhibited satisfying reproducibility, stability, feasibility and fast responses. These results provide a novel way for exact detection of Bilirubin in biotechnology and clinical diagnosis, and broaden the application of graphene and carbon nanotubes in biomaterials. [Copyright &y& Elsevier]

• Multi-element analysis combined with chemometric techniques was used to classify three Codonopsis Radix varieties. • The content of twenty-two elemens in 147 samples from three R. Codonopsis varieties was analyzed. • Twenty-two elements with significant differences could clearly distinguish three R. Codonopsis varieties. • The correct classification rate of linear discriminant analysis was 98.6 %. • Environment with less AAP and lower RH was more conducive for cultivation of high-quality R. Codonopsis. The feasibility of using chemometric techniques associated with multi-element analysis for the classification of three varieties of Codonopsis Radix samples was investigated, and the correlation between environmental factors and metal element content was further explored. It was found that R. Codonopsis was a good source of potassium (average content of 9420 μg/g), magnesium (average content of 1590 μg/g), calcium (average content of 388 μg/g) and iron (average content of 333 μg/g). The statistical analysis results indicated that twenty-two elements content (K, Ca, Mg, Fe, Se, Na, Zn, V, Cu, Cr, Mn, Ni, Co, Sr, Pb, Cd, As, Hg, Al, Ba, Tl and U) were significantly different among three R. Codonopsis varieties, hence it was possible to discriminate R. Codonopsis according to their varieties by LDA (correct classification rate of 98.6 % and cross-validation rate of 95.9 %). In addition, the correlation analysis results indicated that the relative humidity showed significantly negative correlation with essential elemental content such as K, Mg, Ca and Se (correlation coefficients ranged from -0.321 to -0.564, p < 0.05); the annual average precipitation exhibited significantly positive correlation with toxic elemental elements such as As, Cd, Pb and Hg (correlation coefficients ranged from 0.266 to 0.897, p < 0.05). [ABSTRACT FROM AUTHOR]

Abstract: A simple and environmentally friendly synthetic route for the preparation of gelatin functionalized graphene nanosheets (gelatin-GNS) was reported by using exfoliated graphene oxide as a precursor, in which gelatin acted as not only a reducing reagent but also a functionalization reagent to guarantee good dispersibility and stability of the GNS in distilled water and various physiological solutions. The obtained biocompatible gelatin-GNS attaching methotrexate (MTX) via strong π-π stacking interaction, exhibited a high drug loading capacity of MTX and excellent ability for controlled drug release. The pH-dependent release behavior of MTX from MTX@gelatin-GNS showed that the release amount under acid conditions is much higher than that under neutral conditions, which experienced a gelatin-mediated sustained release process. From the cytotoxicity assay, we can see that the MTX@gelatin-GNS showed remarkable toxicity while the gelatin-GNS showed nontoxic at appropriate concentration, both of them might be taken up by A549 cells through a nonspecific endocytosis process. The prepared nanohybrids system offers a novel formulation that combines the unique properties of a biodegradable material, gelatin, and graphene for biomedical applications. Therefore, the gelatin-GNS with good stability and biocompatibility can be selected as an ideal drug carrier to be applied in biomedicine studies. [Copyright &y& Elsevier]

Abstract: A water-soluble sulfated glucan, SHG, was isolated from Hedysarum polybotrys Hand.-Mazz using anion-exchange and gel-permeation chromatography. Elemental analysis indicated that SHG was a sulfated polysaccharide containing small amount of sulfate groups (1.47mass%). The molecular weight was estimated to be 1.72×105 Da by high-performance gel permeation chromatography (HPGPC) and size exclusion chromatography–multi angle laser light scattering (SEC–MALLS). SHG took random coil compact conformation in 0.1M NaNO3. Compositional analysis revealed that SHG was composed of glucose only. On the base of partial acid hydrolysis, methylation analysis, gas chromatography–mass spectrometry (GC–MS), Fourier transform infrared (FT-IR) spectra, nuclear magnetic resonance (NMR) spectroscopy of 1H, 13C, β-correlation spectroscopy (β-COSY), total correlation spectroscopy (TOCSY), heteronuclear single quantum coherence (HSQC), and heteronuclear multiple bond correlation (HMBC), the results showed that SHG was composed of α-d-(1→4)-linked glucopyranosyl residues, with branches at C-6 consisting of non-reducing terminal approximately every eight residues. Sulfate groups may attach to the backbone at O-6, occasionally occurring per thirty-eight anhydrous glucose units. [Copyright &y& Elsevier]

Cooling the ischemic heart by just a few degrees protects it from infarction without affecting its mechanical function, but the mechanism of this protection is unknown. We investigated whether signal transduction pathways might be involved in the anti-infarct effect of mild hypothermia (35°C). Isolated rabbit hearts underwent 30 min of coronary artery occlusion/2 h of reperfusion. They were either maintained at 38.5°C or cooled to 35°C just before and only during ischemia. Infarct size was measured. The effects of the protein kinase C inhibitor chelerythrine, the nitric oxide synthase inhibitor N-nitro- l-arginine methyl ester ( l-NAME), the phosphatidylinositol 3-kinase antagonist wortmannin, or either of the mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitors PD98059 or U0126 on cooling's protection were examined. Myocardial ATP assays were performed and the level of phosphorylation of extracellular signal-regulated kinase (ERK) and MEK was examined by western blotting. To investigate an effect of cooling on protein phosphatase (PPase), a PPase inhibitor cantharidin was tested in the infarct model and the effect of mild hypothermia on PP2A activity in vitro was measured. Infarct size was 34.4 ± 2.2% of the ischemic zone in normothermic (38.5°C) hearts, but only 15.6 ± 8.7% in hearts cooled to 35°C during ischemia. Mechanical function was unaffected. Neither chelerythrine, l-NAME, nor wortmannin had any effect, but both PD98059 and U0126 completely eliminated protection. Ischemia rather than reperfusion was the critical time when ERK had to be active to realize protection. Phosphorylation of ERK and MEK fell during normothermic ischemia, but during hypothermic ischemia phosphorylation of ERK remained high while that of MEK was increased. Cooling only slightly delayed the rate at which ATP fell during ischemia, and ERK inhibition did not affect that attenuation suggesting ATP preservation was unrelated to protection. Cantharidin, like cooling, also protected during ischemia but not at reperfusion, and its protection was dependent on ERK phosphorylation. However, mild hypothermia had a negligible effect on PP2A activity in an in vitro assay. Hence, mild hypothermia preserves ERK and MEK activity during ischemia which somehow protects the heart. While a PPase inhibitor mimicked cooling's protection, a direct effect of cooling on PP2A could not be demonstrated. [ABSTRACT FROM AUTHOR]

• ZIF-8@N-Gr with excellent performance was prepared. • ZIF-8@N-Gr can be used in immunosensor development for Apo-A4 detection. • Lower limit of detection and wider linear range than ELISA kit. • Determination of Apo-A4 in plasma with no need for sample treatment. The fabrication of electrochemical biosensors to directly, rapidly and ultrasensitively detect disease markers in urine or blood samples has become a new and competitive challenge in the field of sensor research. In this paper, a novel electrochemical immunosensor with high selectivity and sensitivity for the detection of the depression marker human apolipoprotein A4 (Apo-A4) was successfully constructed using zeolite imidazole ester metal organic skeleton-nitrogen doped graphene composites (ZIF-8@N-Gr). To this end, because of the higher surface area and biocompatibility, ZIF-8 with abundant biomolecular binding sites provided a good microenvironment for effectively immobilizing antigens. ZIF-8@N-Gr presented a flake structure, as the electrode displayed excellent electrical conductivity, which enhanced the electron transfer and significantly amplified the current signal of the immunosensor. More importantly, these immunosensors are capable of assaying human apolipoprotein A4 (Apo-A4) in 100% serum without suffering from any significant biological interference. Under optimized experimental conditions, the sensor was used for the analysis of whole serum samples and presented a wide linear range from 1.47 × 10−10 g/mL to 3.00 × 10−7 g/mL with a low detection limit of 8.33 × 10−11 g/mL (3σ, n = 15). The satisfactory results of human serum sample analysis indicated that the proposed immunosensor had promising potential in the clinical diagnosis of depression. [ABSTRACT FROM AUTHOR]

• A method for screen toxic and active material basis of Aconitum sinomontanum Nakai was developed based on biological activity guidance coupled with UPLC-Q/TOF-MS technology. • A. sinomontanum chloroform extract possesses significant antinociceptive and anti-inflammatory activities and exhibits significant toxic effects. • The phytochemical analysis of A. sinomontanum chloroform extract indicated the presence of alkaloids. As a result, 30 compounds were detected in A. sinomontanum chloroform extract. A total of 23 compounds, including lappaconine, ranaconidine, 8- O -acetylexcelsine, finaconitine, lappacontine, N -dacetyllappaconitine and ranaconitine, were identified. • Phytochemical analysis indicated that some alkaloids may be the main bioactive ingredient responsible for the toxicity and efficacy of A. sinomontanum chloroform extract. • This work contributes to the determination of the safety of the medicinal use of A. sinomontanum roots. As a folk medicine, Aconitum sinomontanum Nakai (Ranunculaceae, Gaowutou, in Chinese) is used by traditional healers to treat many disorders, including pain and inflammatory diseases, but it exhibits the toxic side effects. This study aimed to obtain toxic extract parts from A. sinomontanum roots and to further evaluate the antinociceptive and anti-inflammatory effects of toxic extract parts on mice. This work also aimed to identify various chemical compositions of the toxic and active extract parts and evaluate the safety profile of this plant. Experimental drugs (petroleum ether, chloroform, ethyl acetate, n-butanol, alcohol and water extracts) were obtained through systematic solvent extraction from 95 % ethanol extract from A. sinomontanum roots. An acute toxicity test was conducted to compare the toxicity of different extracts administered at the maximum dose to screen a highly toxic extract. In pharmacodynamic activity analysis, the antinociceptive activity of the A. sinomontanum toxic extract was assessed using an acetic acid-induced abdominal writhing model and a hot plate test. Anti-inflammatory activity was assessed in terms of xylene-induced inflammation. Ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) was performed to establish a chromatographic fingerprint and to identify various chemical components of the toxic and active extract. Chloroform, water and n-butanol extracts elicited significant toxic effects and had LD 50 of 89.65, 1805.40 and 24409.41 mg/kg, respectively. Antinociceptive and anti-inflammatory activities indicated that the chloroform extract significantly alleviated (p < 0.01) the pain induced by acetic acid with an inhibition rate of 44.7 % (5.9 mg/kg) and 50.4 % (17.7 mg/kg). The chloroform extract also significantly (p < 0.01) increased the latency time during the hot plate test. The latency time at 5.9 and 17.7 mg/kg increased from 15.6 ± 4.1 s to 47.3 ± 6.4 s and from 16.3 ± 3.8 s to 49.8 ± 7.6 s (p < 0.01), respectively, 2 h after treatment. In the inflammatory test, the chloroform extract significantly reduced (p < 0.01) the xylene-induced mouse ear oedema with an inhibition rate of 45.48 % (5.9 mg/kg) and 51.46 % (17.7 mg/kg), respectively. This result indicated that A. sinomontanum chloroform extract was also the active extract part of A. sinomontanum. Phytochemical analysis revealed the presence of alkaloids in the chloroform extract. A total of 30 compounds were detected, and 23 compounds, including lappaconine, ranaconidine, 8- O -acetylexcelsine, sinomontanine H, finaconitine, lappacontine, N -dacetyllappaconitine, ranaconitine and isolappaconitine, were identified. A. sinomontanum chloroform extract possesses antinociceptive and anti-inflammatory activities and exhibits significant toxic effects. Phytochemical analysis indicated that some alkaloids may be the main bioactive ingredient responsible for the toxicity and efficacy of A. sinomontanum. This work contributes to the determination of the safety of the medicinal use of A. sinomontanum roots. [ABSTRACT FROM AUTHOR]

As a new type of conductive material, polyaniline functionalized graphene quantum dots (PAGD), which were prepared by in-situ polymerization had been used to construct a novel electrochemical immunosensor for early screening of depression markers-heat shock protein 70 (HSP70). Profiting from the huge specific surface area, good bioactivity and excellent structure of PAGD, a variety of heat shock protein 70 (HSP70) was firmly loaded on the surface of PAGD for successful construction of basic electrode (HSP70/PAGD/GCE), which was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), respectively. Due to the HSP70 fixed on the surface of basic electrode and the HSP70 in the samples can competitively combine with the horseradish peroxidase labeled human HSP70 antibody (HRP-Strept-Biotin-Ab). As a result, there is negative correlation between the concentration of HSP70 in samples and the detection signal of the proposed electrochemical immunosensor (HRP-Strept-Biotin-Ab-HSP70/PAGD/GCE) in the test liquid. Under conditions optimized for determining HSP70, wide linearity was obtained in the range of 0.0976–100 ng/mL, with a low detection limit of 0.05 ng/mL at 3σ. Moreover, the proposed electrochemical immunosensors was successfully applied to detect HSP70 in plasma samples, and exhibited good precision, acceptable stability and reproducibility. Therefore, this study provides a novel and convenient method for early clinical screening of depression markers, and also provides a reliable and objective analysis method for the diagnosis of depression at the molecular level. Unlabelled Image • The polyaniline functionalized graphene quantum dots (PAGD) was successfully prepared. • The HRP-Strept-Biotin-Ab-HSP70/PAGD/GCE immunosensor for determination of HSP70 was successfully fabricated. • The proposed immunosensor exhibited a broad detection range and low detection limit for HSP70. • It provides a novel and convenient method for early clinical screening of depression markers-HSP70 in real samples. [ABSTRACT FROM AUTHOR]