Globular adiponectin induces esophageal adenocarcinoma cell pyroptosis via the miR-378a-3p/UHRF1 axis.

BACKGROUND: Antiapoptosis is a major factor in the resistance of tumor cells to chemotherapy and radiotherapy. Thus, activation of cell pyroptosis may be an effective option to deal with antiapoptotic cancers such as esophageal adenocarcinoma (EAC). METHODS: Differential expression of ubiquitin-like versus PHD and ring finger structural domain 1 (UHRF1) in EAC and near normal tissues was analyzed, as well as the prognostic impact on survival in EAC. Also, the same study was done for globular adiponectin (gAD). Simultaneously, the mRNA expression of UHRF1 was observed in different EAC cell lines. Real time cellular analysis (RTCA) was used to detect cell proliferation, and flow cytometry and inverted fluorescence microscopy were used to detect pyroptosis. Biocredit analysis was conducted to observe the correlation between UHRF1 and key pyroptosis proteins. OD values and CCK8 assay were used to determine the effect of miR-378a-3p on EAC cells. Quantitative real-time polymerase chain reaction and Western blot were used to detect the correlation between UHRF1, gAD, and miR-378a-3p in EAC cells. Moreover, in vivo and in vitro experiments were performed to detect the relevant effects on tumor migration and invasion after inhibiting UHRF1 expression. RESULTS: UHRF1 was negatively correlated with the survival of patients with EAC, while miR-378a-3p showed the opposite effect. Additionally, gAD promoted EAC cell pyroptosis, upregulated miR-378a-3p, and significantly inhibited the proliferation of EAC cells. gAD directly reduced UHRF1 expression in EAC cells by upregulating miR-378a-3p. In cell migration and invasion assays, inhibition of UHRF1 expression significantly suppressed EAC cell metastasis. In animal experiments, we again demonstrated that gAD induced pyroptosis in EAC cells by inhibiting the expression of UHRF1. CONCLUSION: gAD-induced upregulation of miR-378a-3p significantly inhibited the proliferation of EAC by targeting UHRF1. Therefore, gAD may serve as an alternative therapy for chemotherapy- and radiation-refractory EAC or other cancers with the same mechanism of pyroptosis action.

Coordination of pinna, petiole, and root anatomical traits in 24 tropical-subtropical fern species.

Ferns are primitive vascular plants with diverse morphologies and structures. Plant anatomical traits and their linkages can reflect adaptation to the environment; however, these remain are still poorly understood in ferns. The main objective of this study was to explore whether there was structural coordination among and within organs in fern species. We measured 16 hydraulically related anatomical traits of pinnae, petioles, and roots of 24 representative fern species from the tropical and subtropical forest understory and analyzed trait correlation networks. In addition, we examined phylogenetic signals for the anatomical traits and analyzed co-evolutionary relationships. These results indicated that stomatal density and all petiole anatomical traits exhibited significant phylogenetic signals. Evolutionary correlations were observed between the tracheid diameter and wall thickness of the petiole and between the water transport capacity of the petiole and stomatal density. Conversely, anatomical traits of roots (e.g., root diameter) showed no phylogenetic signals and were not significantly correlated with those of the pinnae and petioles, indicating a lack of structural coordination between the below- and above-ground organs. Unlike angiosperms, vein density is unrelated to stomatal density or pinna thickness in ferns. As root diameter decreased, the cortex-to-stele diameter ratio decreased significantly (enhanced water absorption) in angiosperms but remained unchanged in ferns. These differences lead to different responses of ferns to climate change and improve our knowledge of the water adaptation strategies of ferns.

Regulation of ovalbumin allergenicity and structure-activity relationship analysis based on pulsed electric field technology.

The study focused on the regulation of ovalbumin (OVA) allergenicity using pulsed electric field (PEF) technology and examined the structure-activity link. Following PEF treatment, the ability of OVA to bind to IgE and IgG1 at 6 kHz was inhibited by 30.41 %. According to the microstructure, PEF caused cracks on the OVA surface. Spectral analysis revealed a blue shift in the amide I band and a decrease in α-helix and ß-sheet content indicating that the structure of OVA was unfolded. The disulfide bond conformation was transformed and the structure tended to be disordered. The increased fluorescence intensity indicated that tryptophan and tyrosine were exposed which led an increase in hydrophobicity. In addition, the results of molecular dynamics (MD) simulations confirmed that the stability of OVA was reduced after PEF, which was related to the reduction of hydrogen bonding and the sharp fluctuation of aspartic acid. Therefore, PEF treatment induced the exposure of hydrophobic amino acids and the transformation of disulfide bond configuration which in turn masked or destroyed allergenic epitopes, and ultimately inhibited OVA allergenicity. This study provided insightful information for the production of hypoallergenic eggs and promoted the use of PEF techniques in the food field.

Identification of three lncRNA-related prognostic signatures in gastric cancer by integrated multi-omics analysis.

Aims: The systematic identification of molecular features correlated with the clinical status of gastric cancer (GC) in patients is significant, although such investigation remains insufficient. Methods: GC subtyping based on RNA sequencing, copy number variation and DNA methylation data were derived from The Cancer Genome Atlas program. Prognostics lncRNA biomarkers for GC were identified by univariate Cox, LASSO and SVM-RFE analysis. Results: Three molecular subtypes with significant survival discrepancies, and their specific DEmRNAs and DElncRNAs were identified. Three reliable prognostic-associated lncRNA, including LINC00670, LINC00452 and LINC00160, were selected for GC. Conclusion: Our findings expanded the understanding on the regulatory network of lncRNAs in GC, providing potential targets for prognosis and treatment of GC patients.

Enhancing gel behavior of yellow croaker surimi by fruit extracts: Physicochemical properties and molecular mechanism.

The aim of this study was to investigate the effects of grape seed extract (GSE), acerola cherry extract (ACE), and blueberry extract (BBE) on the physicochemical properties and structure of the yellow croaker surimi gel. In addition, molecular docking and molecular dynamics (MD) simulation were utilized to study the binding mechanism of yellow croaker's fibrillin and fruit extracts. Surimi gel with 1.5% GSE, ACE, and BBE had the highest water holding capacity, hardness, chewability, cohesion, breaking force, breaking distance, gel strength, and densest 3D network structure, according to the experiment's findings. Nevertheless, the cross-linking of proteins in surimi was blocked with the further increase of fruit extract (1.5%-2.0%), and the existing network of surimi was weakened or even destroyed. Three fruit extracts had little effect on the secondary structure of the surimi gel. Besides, hydrophobic and disulfide bonds are the main chemical bonds of croaker surimi. Molecular docking showed that B-type procyanidine (BP) interacted with ASN-183, SER-571, ASP-525, ARG-350, LYS-188, GLU-349, CYS-353, and other active amino acids in croaker protein. Moreover, it can form strong hydrogen bond interaction with ASN-183, SER-571, ASP-525, and ARG-350 at the active sites of protein. The BP-Larimichthys crocea protein system's MD simulation was carried out, and calculations for the simulation's root mean square deviation, root mean square fluctuation, radius of gyration, solvent accessible surface area, and hydrogen bonds were made. It was found that these indices can demonstrate that the BP binding contributes to the stability of the yellow croaker structure.

YKL-40 promotes chemokine expression following drug-induced liver injury via TF-PAR1 pathway in mice.

Background: The inflammatory factor YKL-40 is associated with various inflammatory diseases and is key to remodeling inflammatory cells and tissues. YKL-40 (Chi3l1) promotes the activation of tissue factor (TF), leading to intrahepatic vascular coagulation (IAOC) and liver injury. TF is a key promoter of the exogenous coagulation cascade and is also involved in several signaling involving cell proliferation, apoptosis, charring, migration and inflammatory diseases pathways. However, the effect of YKL-40-induced TF-PAR1 pathway on the expression of downstream chemokines remains unknown. Methods: We established a liver injury model using Concanavalin A (ConA) in C57 BL/6 mice. By adopting various experimental techniques, the effect of YKL-40 induced TF-PAR1 pathway on the expression of downstream chemokine ligand 2 (CCL2) and IP-10 was verified. Results: We found that overexpression of YKL-40 increased the expression of TF, protease-activated receptor 1 (PAR1), CCL2 and IP-10 in mice and exacerbated the severity of liver injury. However, blocking the expression of TF significantly reversed the extent of liver injury. Conclusion: We found that YKL-40 promotes the expression of downstream chemokines ligand 2 (CCL2) and IP-10 by activating the TF-PAR1 pathway, leading to increased recruitment of inflammatory cells and exacerbating the progression of liver injury. This provides a new approach for the clinical treatment of drug-induced liver injury.

Corn Silk Flavonoids Ameliorate Hyperuricemia via PI3K/AKT/NF-κB Pathway.

Hyperuricemia (HUA) is a widespread metabolic disease marked by an elevated level of uric acid, and is a risk factor for premature death. The protective effect of corn silk flavonoids (CSF) against HUA and its potential mechanisms were explored. Five important apoptosis and inflammation-related signaling pathways were identified by network pharmacological analysis. The CSF exhibited significant uric acid (UA)-lowering activity in vitro by decreasing xanthine oxidase (XOD) and increasing hypoxanthine-guanine phosphoribosyl transferase levels. In a potassium oxonate-induced HUA in vivo, CSF treatment effectively inhibited XOD activity and promoted UA excretion. Furthermore, it decreased the levels of TNF-α and IL-6 and restored pathological damage. In summary, CSF is a functional food component to improve HUA by reducing inflammation and apoptosis through the down-regulating PI3K/AKT/NF-κB pathway.

Identification of key immune genes of osteoporosis based on bioinformatics and machine learning.

Introduction: Immunity is involved in a variety of bone metabolic processes, especially osteoporosis. The aim of this study is to explore new bone immune-related markers by bioinformatics method and evaluate their ability to predict osteoporosis. Methods: The mRNA expression profiles were obtained from GSE7158 in Gene expression Omnibus (GEO), and immune-related genes were obtained from ImmPort database (https://www.immport.org/shared/). immune genes related to bone mineral density(BMD) were screened out for differential analysis. protein-protein interaction (PPIs) networks were used to analyze the interrelationships between different immune-related genes (DIRGs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of DIRGs function were performed. A least absolute shrinkage and selection operation (LASSO) regression model and multiple Support Vector Machine-Recursive Feature Elimination (mSVM-RFE) model were constructed to identify the candidate genes for osteoporosis prediction The receiver operator characteristic (ROC) curves were used to validate the performances of predictive models and candidate genes in GEO database (GSE7158,GSE13850).Through the RT - qPCR verify the key genes differentially expressed in peripheral blood mononuclear cells Finally, we constructed a nomogram model for predicting osteoporosis based on five immune-related genes. CIBERSORT algorithm was used to calculate the relative proportion of 22 immune cells. Results: A total of 1158 DEGs and 66 DIRGs were identified between high-BMD and low-BMD women. These DIRGs were mainly enriched in cytokine-mediated signaling pathway, positive regulation of response to external stimulus and the cellular components of genes are mostly localized to external side of plasma membrane. And the KEGG enrichment analysis were mainly involved in Cytokine-cytokine receptor interaction, PI3K-Akt signaling pathway, Neuroactive ligand-receptor interaction,Natural killer cell mediated cytotoxicity. Then five key genes (CCR5, IAPP, IFNA4, IGHV3-73 and PTGER1) were identified and used as features to construct a predictive prognostic model for osteoporosis using the GSE7158 dataset. Conclusion: Immunity plays an important role in the development of osteoporosis.CCR5, IAPP, IFNA4, IGHV3-73 and PTGER1were play an important role in the occurrences and diagnosis of OP.

Mechanism of Targeted Regulation of Ovalbumin Epitopes by Pulsed Electric Field-Assisted Alcalase Treatment.

Egg is one of the eight major food allergens, and ovalbumin (OVA) is the most abundant allergenic protein in eggs. In this study, the effects of pulsed electric field (PEF)-assisted Alcalase hydrolysis on the spatial conformation and potential allergenicity of OVA were studied, and the mechanism of its inhibiting allergic reactions effect was revealed. PEF-assisted Alcalase hydrolysis increased the degree of hydrolysis, surface hydrophobicity, and free sulfhydryl group content. Moreover, the reduction in the α-helix content, fluorescence intensity, and disulfide bond content suggested that PEF promoted the OVA hydrolysis by Alcalase. Additionally, enzyme-linked immunosorbent assay data indicated that PEF-assisted Alcalase hydrolysis hindered OVA binding to immunoglobulins E and G1. Finally, based on bioinformatics combined with mass spectrometry, PEF-assisted Alcalase reduced OVA-induced allergic reactions by destroying epitopes in OVA. Overall, PEF technology further destroyed the epitopes of allergens by targeting the binding sites of substrates and enzymes to improve the affinity of enzymes and substrates, reducing allergic reactions.

Enhanced effect and mechanism of nano Fe-Ca bimetallic oxide modified substrate on Cu(II) and Ni(II) removal in constructed wetland.

In this study, Fe2O3 nanoparticles (Fe2O3 NPs) and CaO NPs were loaded on the zeolite sphere carrier to create nano Fe-Ca bimetallic oxide (Fe-Ca-NBMO) modified substrate, which was introduced into constructed wetland (CW) to remove Cu(II) and Ni(II) via constructing "substrate-microorganism" system. Adsorption experiments showed that the equilibrium adsorption capacities of Fe-Ca-NBMO modified substrate for Cu(II) and Ni(II) were respectively 706.48 and 410.59 mg/kg at an initial concentration of 20 mg/L, 2.45 and 2.39 times of gravel. The Cu(II) and Ni(II) removal efficiencies in CW with Fe-Ca-NBMO modified substrate respectively reached 99.7% and 99.9% at an influent concentration of 100 mg/L, significantly higher than those in gravel-based CW (47.0% and 34.3%). Fe-Ca-NBMO modified substrate could promote Cu(II) and Ni(II) removal by increasing electrostatic adsorption, chemical precipitation, as well as the abundances of resistant microorganisms (Geobacter, Desulfuromonas, Zoogloea, Dechloromonas, and Desulfobacter) and functional genes (copA, cusABC, ABC.CD.P, gshB, and exbB). This study provided an effective method to enhance Cu(II) and Ni(II) removal of electroplating wastewater by CW with Fe-Ca-NBMO modified substrate.

Bio-clogging mitigation in constructed wetland using microbial fuel cells with novel hybrid air-photocathode.

Excessive accumulation of extracellular polymeric substances (EPS) in constructed wetland (CW) substrate can lead to bio-clogging and affect the long-term stable operation of CW. In this study, a microbial fuel cell (MFC) was coupled with air-photocathode to mitigate CW bio-clogging by enhancing the micro-electric field environment. Because TiO2/biochar could catalyze and accelerate oxygen reduction reaction, further promoting the gain of electric energy, the electricity generation of the tandem CW-photocatalytic fuel cell (CW-PFC) reached 90.78 mW m-3. After bio-clogging was mitigated in situ in tandem CW-PFC, the porosity of CW could be restored to about 62.5 % of the initial porosity, and the zeta potential of EPS showed an obvious increase (-14.98 mV). The removal efficiencies of NH4+-N and chemical oxygen demand (COD) in tandem CW-PFC were respectively 31.8 ± 7.2 % and 86.1 ± 6.8 %, higher than those in control system (21.1 ± 11.0 % and 73.3 ± 5.6 %). Tandem CW-PFC could accelerate the degradation of EPS into small molecules (such as aromatic protein) by enhancing the electron transfer. Furthermore, microbiome structure analysis indicated that the enrichment of characteristic microorganisms (Anaerovorax) for degradation of protein-related pollutants, and electroactive bacteria (Geobacter and Trichococcus) promoted EPS degradation and electron transfer. The degradation of EPS might be attributed to the up-regulation of the abundances of carbohydrate and amino acid metabolism. This study provided a promising new strategy for synergic mitigation and prevention of bio-clogging in CW by coupling with MFC and photocatalysis.

Biomarkers for chemotherapy and drug resistance in the mismatch repair pathway.

Drugs targeting DNA repair have developed rapidly in cancer therapy, and numerous inhibitors have already been utilized in preclinical and clinical stages. To optimize the selection of patients for treatment, it is essential to discover biomarkers to anticipate chemotherapy response. The DNA mismatch repair (MMR) pathway is closely correlated with cancer susceptibility and plays an important role in the occurrence and development of cancers. Here, we give a concise introduction of the MMR genes and focus on the potential biomarkers of chemotherapeutic response and resistance. It has been clarified that the status of MMR may affect the outcome of chemotherapy. However, the specific underlying mechanisms as well as contradictory results continue to raise considerable controversy and concern. In this review, we summarize the current literature to provide a general overview.

Efficacy of Baishao Luoshi decoction on synaptic plasticity in rats with post stroke spasticity.

OBJECTIVE: To evaluate the efficacy of Baishao Luoshi decoction (, BD) on synaptic plasticity in rats with post stroke spasticity (PSS), and to study the mechanism behind the action. METHODS: The PSS model of rat was established by middle cerebral artery occlusion (MCAO). The neurological deficit symptoms were evaluated by modified neurological deficit score (mNSS). Muscle tension were evaluated by Modified Ashworth score (MAS). Transmission electron microscopy (TEM) was used to observe the synaptic ultrastructure. The expression of synaptic plasticity-related protein brain derived neurotrophic factor (BDNF), growth associated protein-43 (GAP43), synaptophysin (p38) and microtubule-associated protein 2 (MAP2) in the brain tissue around the infarct were detected by Western blotting. RESULTS: We found that mNSS were significantly improved and limb spasticity was ameliorated treated by BD. The thickness of postsynaptic density and the synaptic curvature increased significantly. The expression of synaptic plasticity-related protein BDNF, GAP43, p38, MAP2 in the brain tissue around the infarct were raised remarkably after treated by BD. CONCLUSIONS: Alleviating PSS by BD may be related to rescuing the synaptic plasticity, which provides a probable new therapeutic method for PSS.

Pulsed Electric Field-Assisted Alcalase Treatment Reduces the Allergenicity and Eliminates the Antigenic Epitopes of Ovomucoid.

Physically assisted chemical modifications can effectively reduce the allergenicity of ovomucoid (OVM). However, only a few studies have used pulsed electric field (PEF)-assisted alcalase hydrolysis to reduce the allergenicity of OVM. Herein, we investigated the effect of PEF-assisted alcalase treatment on the spatial conformation, allergenicity, and antigenic epitopes of OVM based on multispectroscopic analyses, bioinformatics, and mass spectrometry. The results showed that PEF-assisted alcalase treatment promoted the hydrolysis of OVM; moreover, the α-helix content and surface hydrophobicity of OVM significantly decreased, which disordered its spatial conformation and weakened its intermolecular interactions. Additionally, enzyme-linked immunosorbent assay (ELISA) results showed that the PEF-assisted alcalase treatment significantly reduced the binding levels of IgE and IgG1, which were 47.66 and 36.41%, respectively. Finally, eight epitopes of OVM were obtained by immunoinformatic tools. Nano-high performance liquid chromatography coupled to tandem mass spectrometry (nano-HPLC MS/MS) results showed that the hydrolysate of OVM and alcalase (HOVM) had nine more peptide-containing epitopes than the hydrolysate of PEF-treated OVM and PEF-treated alcalase (HOVM-PP'), indicating that PEF could promote the elimination of linear epitopes in OVM, thereby reducing OVM allergenicity.

TDDFT study on the simultaneous sensing mechanism for peroxynitrite and glutathione by a bifunctional fluorescent probe.

Using time-dependent density functional theory (TDDFT) method, the response mechanism of a reported bifunctional fluorescent probe for simultaneous recognition of peroxynitrite and glutathione (Chem. Commun. 2018, 54, 11336) was theoretically studied. Calculated vertical excitation energies based on the ground-state and excited-state geometries were consistent with the corresponding experimental ultraviolet-visible and fluorescence spectra. In the ground state, electron delocalization in the probe was limited because its geometry was restrained by steric hindrance. Frontier molecular orbital analysis has shown that the probe should undergo photoinduced electron transfer (PET) from the benzothiazole moiety to the maleimide moiety after excitation. The nonplanar structure together with PET led to fluorescence quenching of the probe. The probe could be dealkylated by peroxynitrite anion. The resulting intramolecular hydrogen bond increasesd the planarity of the molecule, while also gave rise to an excited-state proton-transfer process. Moreover, the addition reaction between the probe and glutathione inhibited the PET process. These two analytes together contributed to the fluorescence enhancement of the final product. This theoretical sensing mechanism for peroxynitrite and glutathione may potentially be important for the design and enhancement of novel probes.

Evaluation of Fourier transform infrared (FTIR) spectroscopy with multivariate analysis as a novel diagnostic tool for lymph node metastasis in gastric cancer.

Fourier transform infrared (FTIR) spectroscopy is a vibration spectroscopy that uses infrared radiation to vibrate to absorb the molecular bonds in its absorbed sample. The main purpose of this study was to evaluate FTIR spectroscopy as a novel diagnostic tool for lymph node metastasis (LNM) of gastric cancer. We collected 160 fresh non-metastatic and metastatic lymph nodes (80 each) from 60 patients with gastric cancer for spectral analysis. FTIR spectra of lymph node (LN) samples were obtained in the wavenumber range of 4000 cm-1 to 900 cm-1. We calculated the changes in the ratio of spectral intensity (/ I1460). Principal component analysis (PCA) and Fisher's discriminant analysis (FDA) were used to distinguish malignant from normal LN. Four significant bands at 1080 cm-1, 1640 cm-1, 1740 cm-1 and 3260 cm-1 separated metastatic and non-metastatic LN spectra into two distinct groups by PCA.T-tests showed that, along with the relative intensity ratios (I1080/I1460, I1640/I1460, I3260/I1460, I1740/I1460), these band ratios were also able to differentiate between malignant and benign LN spectra. Six parameters (P1080 cm-1, P1300 cm-1, I1080/I1460, I1640/I1460, I3260/I1460, I1740/I1460) were selected as independent factors to set up discriminant functions. The sensitivity of FTIR spectroscopy in diagnosing LNM was 95 % by discriminant analysis. Our study suggested that FTIR spectroscopy can be a useful tool to examine LNM with high sensitivity and specificity for LNM diagnosis. Therefore it can be used in clinical practice as a non-invasive method.

Synthesis of Tetrahydro-5H-indolo[2,3-b]quinolines through Copper-Catalyzed Cascade Reactions of Aza-o-quinone Methides with Indoles.

A variety of tetrahydro-5H-indolo[2,3-b]quinolines were prepared in 40-97% yields through a copper(II)-catalyzed cascade reaction of aza-o-quinone methides generated in situ from 2-(chloromethyl)anilines and indoles. Experimental results showed that the reaction underwent double 1,4-additions and sequential intramolecular cyclization. The present method features broad substrate scope, good functional group tolerance, and easy gram scalable preparation of indolo[2,3-b]quinolines.

A rapid multiplex assay of human malaria parasites by digital PCR.

BACKGROUND: Blood smear examination through traditional optical microscopy is the gold standard for malaria diagnosis. However, it imposes strict requirements for operational staff and its sensitivity cannot perfectly satisfy the needs of clinical requirements. More sensitive and accurate modern technologies should be applied to this field. Digital PCR (dPCR), as an absolute quantification detection method, can serve as an effective tool to facilitate the diagnosis and classification of different malaria species. OBJECTIVE: We aimed to establish a new multiplex dPCR detection system for four main Plasmodium species: P. vivax, P. falciparum, P. ovale and P. malariae, which can distinguish exact species of malaria by one PCR reaction. METHODS: A total of 39 patients were identified as malaria-positive by microscopic examination in Huashan Hospital from 2016 to 2021; seventy blood samples from these patients were collected. Additionally, 20 healthy individuals, 20 patients with fever and 6 patients with other types of blood parasites infection were also included in this study. Each blood sample was subjected to examination by both blood smears and dPCR. By optimizing four different fluorescence-labeled probes in one reaction system, dPCR permitted the performance of accurate quantitation and working out the exact number of copies of malaria DNA per microliter in whole blood. Rapid diagnostic tests were also conducted to verify part of the results obtained by dPCR. RESULTS: The dPCR system was able to make rapid diagnosis and quantification of malaria DNA samples. The analytical sensitivity of multiplex dPCR was as low as 0.557 copies/µL (95% CI 0.521 to 0.607), and it had a sensitivity of 98.0% and a specificity of 100% in clinical samples. Additionally, three multiple malaria co-infection samples have been detected by this dPCR system, including one triple malaria infection case. By testing consecutive daily blood samples of Patient 39, dPCR facilitated monitoring the efficacy of drug treatment. It showed that the DNA concentrations of P. falciparum ranged from 5474 copies/µL to 0 copies/µL, which can reflect the efficacy of antimalarials in real time. This study also found that haemocyte samples (plasma removed) rather than whole blood had higher malaria detection capability and an enhanced positive rate. CONCLUSION: The multiplex dPCR system newly established here made a substantial contribution in detecting malaria infection at low concentrations. It is suitable for mixed-infection diagnosis and multi-sample continuous monitoring, and presents a promising candidate as an absolute quantitative tool in clinical practice.

Regulation of action sites for reducing the allergenicity of pea protein based on enzymatic hydrolysis with Alcalase.

Enzymatic hydrolysis could reduce the allergenicity of pea protein. Box-Behnken model was used to extract vicilin with the lowest and highest allergenicity, and enzymatic hydrolysis, electrophoresis, spectroscopy, bioinformatics, and peptidomics of Nano-LC-MS/MS were utilized to explore the relationship between reduced allergenicity and structural changes. After enzymatic hydrolysis, the allergenicity of L-vicilin hydrolysates (L-VHs) and H-vicilin hydrolysates (H-VHs) decreased significantly (P < 0.05). Furthermore, large-molecular-weight subunits in L-vicilin and H-vicilin were decomposed into <11 kDa peptides, and their surface hydrophobicity were increased. The OH, NH, and CO groups underwent stretching vibrations, and α-helix and ß-sheet were transformed into ß-turn and random coils. Additionally, linear epitopes of P13918, D3VND7, D3VNE2, and P02856 in L-VHs and H-VHs were cut into different fragments. Among them, distinct linear epitope fragments of them might be responsible for the difference in their allergenicity. Therefore, enzymatic hydrolysis with Alclase could effectively reduce the allergenicity of vicilin by regulating sensitization sites.

Investigation of pharmacological characteristics and mechanisms of Qufeng Gutong Cataplasm based on a "disease-syndrome-symptom-formula" association network analysis / 药学学报

Growing clinical evidence shows that Qufeng Gutong Cataplasm may exert a significant analgesic effect. However, the pharmacological characteristics and mechanisms underlying this prescription are still unclear. In the current study, a "disease-syndrome-symptom-formula" association network analysis was performed to explore the pharmacological characteristics and mechanisms of Qufeng Gutong Cataplasm against osteoarthritis (OA), neuropathic pain (NP), chronic inflammatory pain (CIP) and myofascial pain syndrome (MPS) by integrating clinical phenomics data, transcriptomics data and biological interaction network mining. As a result, the three functional modules (Qufeng Sanhan-QFSHG, Shujin Huoxue-SJHXG and Xiaozhong Zhitong-XZZTG) enriched by the drug network targets were all related to the pharmacological effects of Qufeng Gutong Cataplasm, including dispersing cold and relieving pain, activating blood and relieving pain, reducing swelling and relieving pain. In addition, the main pharmacological effects of QFSHG and XZZTG were dispelling wind and dispersing cold and dehumidifying, promoting Qi and reducing swelling and relieving pain, respectively. In terms of reversing the imbalance of "immune-inflammation-vascular axis", the main pharmacological effects of SJHXG were regulating the liver and promoting Qi, activating blood circulation and removing stasis. Mechanically, the key network targets of Qufeng Gutong Cataplasm against OA, NP, CIP and MPS may play a therapeutic role in relieving hyperalgesia and paresthesia by reversing the "neuro-endocrine-immune" imbalance system during the occurrence and progression of diseases. In conclusion, our data indicate that Qufeng Gutong Cataplasm may relieve the pain and wind-cold-dampness arthralgia syndrome related symptoms by regulating the "neuro-endocrine-immune" system, neurological and endocrine disorders and reversing the imbalance of "immunity-inflammation". The relevant results may provide a network-based evidence for clinical positioning of Qufeng Gutong Cataplasm, and offer a direction for further clinical and experimental validation.