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Abstract: SARS-CoV-2 is a disease that endangers both human life and the economy. There was an 11- month period of relative evolutionary standstill following the appearance of SARS-CoV-2 in late 2019. However, the emergence of clusters of mutations known as' variants of concern 'with variable viral properties such as transmissibility and antigenicity defined the evolution of SARS-CoV-2. Several efforts have been made in recent months to understand the atomic level properties of SARS-CoV-2. A review of the literature on SARS-CoV-2 mutations is offered in this paper. The critical activities performed by different domains of the SARS-CoV-2 genome throughout the virus's entry into the host and overall viral life cycle are discussed in detail. These structural traits may potentially pave the way for the development of a vaccine and medication to combat the SARS-CoV-2 sickness.

Background: Intestinal microbiota is the primary target for the multifunctional nature of berberine. Berberine can reverse the structure and composition of gut microbiota under pathological conditions. This study aimed to investigate the effects of berberine on uric acid (UA) metabolism and gut microbiota in a hyperuricemia rat model established using potassium oxonate. Methods: Sprague-Dawley (SD) male rats were divided into a normal control group (n= 10), a hyperuricemia group (n = 12) and a berberine-treated group (n = 11). The UA level in serum, urine and fecal, blood xanthine oxidase (XOD), and urate transports ABCG2 and Galectin-9 in the liver and colon, were evaluated using ELISA kits. The alterations in gut microbiota were investigated using 16S rRNA sequencing. Results: The UA level in the hyperuricemia group was significantly elevated (p Conclusion: Berberine might be a possible therapeutic candidate in hyperuricemia, which could regulate UA metabolism by affecting XOD, and urate transports and partly by regulating gut microbiota.

Background: Indole and its derivatives have a wide range of pharmacological effects, including analgesic, antimicrobial, antidepressant, anti-diabetic, anti-convulsant, anti-helminthic, and anti-inflammatory properties. They are crucial structural components of many of today's powerful antioxidant medications. Objective: Using the Schotten–Baumann reaction, the indole ring was linked to other key heterocyclic moieties such as morpholine, imidazole, piperidine, and piperazine at the active 3rd position and then tested for antioxidant activity. Method: Synthesis of derivatives was accomplished under appropriate conditions and characterized by IR, NMR (1H and 13C), and mass spectrum. Using the Swiss ADME online application, ADME properties were also determined. The in vitro antioxidant activity was measured using DPPH and Reducing power method. Results: In the DPPH assay, compounds 5a (IC50=1.01±0.22 μg/mL), 5k (IC50=1.21±0.07μg/mL), whereas compounds 5a (EC50=23±1.00 μg/mL), 5h (EC50=26±2.42 μg/mL) in the reducing power assay were most potent as compared with standard Ascorbic acid. Compounds 5a, 5h, and 5k demonstrated maximal potency equivalent to standard. Lipinski's rule was followed in ADME outcomes. Conclusion: The synthesis and evaluation of indole derivatives to investigate their antioxidant action has received a lot of attention. These discoveries could lead to more effective antioxidant candidates being designed and developed.

Background: Hypoxia-induced pulmonary hypertension (HPH) represents a severe pulmonary disorder with high morbidity and mortality, which necessitates identifying the critical molecular mechanisms underlying HPH pathogenesis. Method: The mRNA expression microarray GSE15197 (containing 8 pulmonary tissues from HPH and 13 normal controls) was downloaded from Gene Expression Omnibus (GEO). Gene ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) were executed by RStudio software. The Protein-Protein Interaction (PPI) network was visualized and established using Cytoscape, and the cytoHubba app from Cytoscape was used to pick out the hub modules. The infiltration of immune cells in HPH was analyzed using the CIBERSORTx. To confirm the potential hub genes, real-time quantitative reverse transcription PCR (qRT-PCR) was conducted using lung tissues of rat HPH models and controls. Results: A total of 852 upregulated and 547 downregulated genes were identified. The top terms in biological processes were apoptosis, proliferation, and regulation of the MAPK cascade, including ERK1/2. Cytoplasm, cytosol, and membrane were enriched in cellular component groups. Molecular functions mainly focus on protein binding, protein serine/threonine kinase activity and identical protein binding. KEGG analysis identified pathways in cancer, regulation of actin cytoskeleton and rap1 signaling pathway. There was significantly different immune cell infiltration between HPH and normal control samples. High proportions of the memory subsets of B cells and CD4 cells, Macrophages M2 subtype, and resting Dendritic cells were found in HPH samples, while high proportions of naive CD4 cells and resting mast cells were found in normal control samples. The qRTPCR results showed that among the ten identified hub modules, FBXL3, FBXL13 and XCL1 mRNA levels were upregulated, while NEDD4L, NPFFR2 and EDN3 were downregulated in HPH rats compared with control rats. Conclusion: Our study revealed the key genes and the involvement of immune cell infiltration in HPH, thus providing new insight into the pathogenesis of HPH and potential treatment targets for patients with HPH.

Background: More than 95% patients with acute promyelocytic leukemia (APL) carry the PML-RARα fusion oncoprotein. Arsenic trioxide (ATO) is an efficacious therapeutic agent for APL, and the mechanism involves the binding with PML and degradation of PML-RARα protein. Pyrrolidine dithiocarbamate (PDTC) demonstrates the function of facilitating the cytotoxic effect of ATO. Purpose: To investigate whether PDTC is potential to enhance the cytotoxic effect of ATO to APL cells by acting on PML-RARα oncoproteins. Methods: Inhibitory effects of drugs on cell viability were examined by CCK-8 test, and apoptosis was evaluated by flow cytometry. Western blotting and immunofluorescence assays were used to explore the mechanism Results: PDTC improved the effect of ATO on inhibiting proliferation of NB4 cells in vitro. Further, PDTC-ATO promoted apoptosis and cell cycle arrest in NB4 cells. The expression of caspase- 3 and Bcl-2 was reduced in PDTC-ATO-treated NB4 cells, while cleaved caspase-3 and Bax was up-regulated. Furthermore, less PML-RARα expression were found in PDTC-ATO-treated NB4 cells than that in NB4 cells treated with ATO singly. Laser confocal microscopy showed that protein colocalization of PML and RARα was disrupted more significantly by PDTC-ATO treatment than that with ATO monotherapy. Conclusions: In conclusion, PDTC enhanced the cytotoxic effect of ATO on APL, and the mechanism was, at least in part, related to the promotion of ATO-induced degradation of PML-RARα fusion protein via forming a complex PDTC-ATO.

Abstract: Fatty liver disease (FLD), which includes both non-alcoholic fatty liver disease (NAFLD) and alcoholic fatty liver disease (ALD), is a worldwide health concern. The etiology of ALD is long-term alcohol consumption, while NAFLD is defined as an abnormal amount of lipid present in liver cells, which is not caused by alcohol intake and has recently been identified as a hepatic manifestation of metabolic syndrome (such as type 2 diabetes, obesity, hypertension, and obesity). Inflammation, oxidative stress, and lipid metabolic dysregulation are all known to play a role in FLD progression. Alternative and natural therapies are desperately needed to treat this disease since existing pharmaceuticals are mostly ineffective. The aldose reductase (AR)/polyol pathway has recently been shown to play a role in developing FLD by contributing to inflammation, oxidative stress, apoptosis, and fat accumulation. Herein, we review the effects of plantderived compounds capable of inhibiting AR in FLD models. Natural AR inhibitors have been found to improve FLD in part by suppressing inflammation, oxidative stress, and steatosis via the regulation of several critical pathways, including the peroxisome proliferator-activated receptor (PPAR) pathway, cytochrome P450 2E1 (CYP2E1) pathway, AMP-activated protein kinase (AMPK) pathway, etc. This review revealed that natural compounds with AR inhibitory effects are a promising class of therapeutic agents for FLD.

Abstract: One of the most dynamic approaches in biotechnology is reverse vaccinology, which plays a huge role in today’s developing vaccines. It has the capability of exploring and identifying the most potent vaccine candidate in a limited period of time. The first successful novel approach of reverse vaccinology was observed in Neisseria meningitidis serogroup B, which has revolutionised the whole field of computational biology. In this review, we have summarized the application of reverse vaccinology for different infectious diseases, discussed epitope prediction and various available bioinformatic tools, and explored the advantages, limitations and necessary elements of this approach. Some of the modifications in the reverse vaccinology approach, like pan-genome and comparative reverse vaccinology, are also outlined. Vaccines for illnesses like AIDS and hepatitis C have not yet been developed. Computer Aided Reverse vaccinology has the potential to be a game-changer in this area. The use of computational tools, pipelines and advanced soft-computing methods, such as artificial intelligence and deep learning, and exploitation of available omics data in integration have paved the way for speedy and effective vaccine designing. Is reverse vaccinology a viable option for developing vaccines against such infections, or is it a myth? Vaccine development gained momentum after the spread of various infections, resulting in numerous deaths; these vaccines are developed using the traditional technique, which includes inactivated microorganisms. As a result, reverse vaccinology may be a far superior technique for creating an effective vaccine.

Background: Abnormal epigenetic alterations influenced by external factors and affecting DNA expression contribute to the development of asthma. However, the role of nasal epithelium in airway inflammation remains unknown. Objective: The objective of this study is to identify novel DNA promoter hypermethylation, which completely suppresses mRNA expression in nasal epithelial of asthma. Methods: Microarray datasets were downloaded from the Gene Expression Omnibus [GEO] database. Gene expression and DNA promoter methylation sites in key correlated modules between asthma and normal were identified by weighted gene co-expression network analysis [WGCNA]. Gene Ontology and KEGG were conducted to analyse the function of genes. Further validation was performed in human BEAS-2B cells challenged by IL-4 or IL-13. Results: Lightcyan, lightgreen, midnightblue, cyan and tan modules in mRNA expression dataset showed a close relationship with asthma, in which genes were enriched in TNF, IL-17, ErbB, MAPK and Estrogen signalling pathways. Blue and turquoise modules in methylation profiling dataset were associated with asthma. 49 lowly expressed genes were identified to be correlated with aberrant DNA hypermethylation of promoters. Among these genes, the mRNA levels of BCL10, GADD45B, LSR and SQSTM1 were downregulated in BEAS-2B cells challenged with IL-4 or IL-13. Conclusion: Four potential genes in nasal epithelium, by hypermethylating their own DNA promoter, might mediate the inflammatory response in the pathogenesis of asthma. Analyzing epigenomic data by integrated bioinformatics helps to understand the role of DNA methylation in asthma, with the goal of providing new perspectives for diagnosis and therapy.

Background: Intervertebral Disc Degeneration (IDD) is a heterogeneous spinal disease whose underlying molecular mechanism is unclear. Objectives: This study aimed to identify, profile, and analyze microRNAs (miRNAs) related to IDD. Method: Microarray Gene Expression IDD data (GSE63492) were downloaded from Gene Expression Omnibus datasets. We employed Weighted Gene Co-Expression Network Analysis (WGCNA) to construct a miRNA co-expression network, and the miRNAs related to the IDD stage were detected. The number of differentially expressed miRNAs between normal and degenerated nucleus pulposus tissues was calculated. Twenty-three clinical specimens were used to validate the expression of miRNAs using qRT-PCR. Results: WGCNA identified 48 miRNAs significantly related to the IDD stage, and 94 miRNAs that were significantly different between normal and degenerated nucleus pulposus tissues. We selected 32 overlapping miRNAs and identified 347 corresponding target genes. The integrative analysis revealed the biological function and pathways of these targeted genes. Analysis of clinical specimens validated that hsa-miR-4534 was upregulated in IDD, whereas hsa-miR-1827 and hsa-miR- 185-5p were downregulated in IDD. Conclusion: This study has identified a subset of miRNAs that are related to IDD pathogenesis and hub miRNAs that are keys to the IDD co-expression network, which may potentially be utilized as indicators for treatment.

Background and Objective: Chronic prostatitis (CP) is one of the most common diseases in young and middle-aged men but lacks effective treatment. Shuangshi Tonglin Capsule (SSTLC) is a clinical drug for the treatment of chronic prostatitis. However, the underlying molecular mechanisms of SSTLC in treating CP are still unclear. In this study, we researched the underlying mechanisms of SSTLC in treating chronic prostatitis. Methods: The ingredients of SSTLC were received from the TCMSP and BATMAN databases, and the CP targets were collected based on GeneCards and OMIM. Then, the PPI network and the “drug-ingredient-target” network map were constructed. GO and KEGG enrichment analyses by using DAVID. Molecular docking was performed by using AutoDock 4.2 and PyMol. And using animal experiments to verify the potential effect of SSTLC in CP. Results: SSTLC contained 10 herbs, 158 chemical ingredients and 277 targets, 2002, diseaserelated targets were obtained. Network analysis outcomes indicated that VEGFA, TNF, MAPK1, EGFR, and MAPK8 are the key targets of SSTLC in treating chronic prostatitis. Furthermore, molecular docking revealed that quercetin, luteolin, and kaempferol exhibited a strong binding effect. Animal experimental indicated that SSTLC can reduce the pathological damage to prostate tissue. And, we found that high-dose SSTLC significantly reduced the level of TNF-α and downregulated the expression of EGFR, p-p38 and p-ERK1/2 (P Conclusions: This study determined the pharmacological effects of SSTLC and the potential mechanism of action on SSTLC to treat CP, it provides a new idea for traditional Chinese medicine to treat chronic prostatitis.

Background: Non-small cell lung cancer (NSCLC) is associated with high morbidity and mortality. Dysregulation of lncRNAs leads to NSCLC progression. Objective: This study aims to explore the regulatory mechanism of lncRNA LINC01234 in NSCLC. Materials and Methods: LINC01234 expression in NSCLC cells was determined. Cell proliferation was detected using CCK-8, colony formation, and EDU assays after transfection of siRNA LINC01234 into H1299 cells and transfection of pcDNA3.1-LINC01234 into H1975 cells. Subcellular localization of LINC01234 was predicted and the binding relations between LINC01234 and miR-433-3p as well as miR-433-3p and GRB2 were verified. The expression levels of miR-433-3p and GRB2 in NSCLC cells were determined. Joint experiments of miR-433-3p inhibitor + si- LINC01234-1 or oe-GRB2 + si-LINC01234-1 were conducted to verify the role of miR-433-3p and GRB2 in NSCLC cell malignant proliferation. Results: LINC01234 was abundantly expressed in NSCLC cells. LINC01234 silencing reduced NSCLC cell proliferation while LINC01234 overexpression enhanced cell proliferation. LINC01234 competitively bound to miR-433-3p and miR-433-3p directly targeted GRB2. miR- 433-3p knockdown or GRB2 overexpression counteracted the repressive effect of LINC01234 silencing on NSCLC cell malignant proliferation. Conclusion: LINC01234 competitively bound to miR-433-3p and promoted GRB2 transcription to augment NSCLC cell malignant proliferation.

Background: Potatoes are extremely important compared to other vegetable crops. Several species of fungi cause severe damage to different components of the plant (leaves, stems, and tubers), leading to significant losses during cultivation and even after harvest. In the framework of the investigation for alternative methods against the proliferation of these fungi, the present work focuses on the study of the antifungal effect of essential oils of some plants that could be used to solve these problems without the use of harmful chemical substances. Aim and Purpose: This study aims to discuss the chemical composition of essential oils of Artemisia herba alba and Ammoides verticillata and evaluate their in vitro and in vivo antifungal activities in order to prevent fungal diseases of potatoes and replace chemical pesticides that cause neurodegenerative diseases. Methods:: Essential oils extracted from the aerial parts of the plants Artemesia herba alba and Ammoides verticillata were analyzed by gas chromatography-mass spectrometry (GC/MS) and tested individually and in combination for their antifungal effects against Fusarium solani, Penicillium expansum, and Aspergillus flavus, by the radial growth technique. Results: The essential oil of A. herba alba was mainly composed of hydrocarbon monoterpenes (80.8%), while the oil of A. verticillata was mainly composed of oxygenated monoterpenes (54.4%). The study on the antifungal effect of essential oils in vitro showed that essential oil of A. Verticillata was more effective against P. expansum (64.40%) than A. flovus (41.10%) and F. solani (53.30%), and the oil of A.herba alba was more effective against A. flavus (54.40%) and (42%) F. Solani. While the combination of these two essential oils of A. verticillata and A. herba-alba gave excellent results, i.e., 100% against P. expansum and A. flavus and 94.40% against F. solani and provided an in vivo protection to the potato in the range of 80% to 90% against the three fungi. Conclusion: A. herba alba and A. verticillata individual and combined essential oils are very effective antifungal biocides that can be used as an alternative to chemical pesticides to prevent their harmful effects on health.

Background: Traditional Chinese medicine formula (TCMF) Run-zao-zhi-yang capsule (RZZY) is commonly used in treating itch in China. However, there are few studies on its mechanisms. In this study, we revealed the mechanisms and molecular targets of RZZY for itch by network pharmacology, molecular docking, and in vitro experiments. Methods: The network pharmacology consisted of active ingredient collection, target acquisition, enrichment analysis, biological process analysis, and network construction. Molecular docking was carried out using molegro virtual docker (MVD) software. LPS-induced RAW 264.7 cells were used to evaluate the in vitro anti-inflammatory activity. Results: We collected 483 high-confidence targets that interacted with 16 active compounds of RZZY, including 121 common genes related to itch. 43 important targets and 20 important pathways were identified according to the network and system analysis. Target-pathway network function analysis suggested that RZZY is treated for itch by multiple ways in immune regulation, hormone adjustment, anti-inflammation, and anti-oxidation. Molecular docking results demonstrated that daidzein and formononetin could be closely combined with 4 proteins. In vitro experiments displayed that RZZY, sophocarpine, catalpol, emodin, and daidzein had suppressive effects against TNF-α, IL-1β, or IL-6 production in LPS-induced RAW 264.7 cells. Interestingly, the result of network pharmacology revealed that RZZY might be more suitable for senile pruritus, consistent with the bibliometric analysis of RZZY’s clinical indications. Conclusions: This study illustrated the potential mechanisms and molecular targets of RZZY for itch, which may contribute to the proper use of RZZY in clinical practice.

Background and Aim: Major Depressive Disorder (MDD) is a common affective disorder. GuiPi decoction (GPD) is used to treat depression in China, Japan, and Korea. However, its effective ingredients and antidepressant mechanisms remain unclear. We attempted to reveal the potential mechanisms of GPD in the treatment of MDD by network pharmacology and molecular docking. In addition, we conducted an enzymatic activity assay to validate the results of molecular docking. Methods: GPD-related compounds and targets, and MDD-related targets were retrieved from databases and literature. The herb-compound-target network was constructed by Cytoscape. The protein- protein interaction network was built using the STRING database to find key targets of GPD on MDD. Enrichment analysis of shared targets was analyzed by MetaCore database to obtain the potential pathway and biological process of GPD on MDD. The main active compounds treating MDD were screened by molecular docking. The PDE4s inhibitors were screened and verified by an enzyme activity assay. Results: GPD contained 1222 ingredients and 190 potential targets for anti-MDD. Possible biological processes regulated by GPD were neurophysiological processes, blood vessel morphogenesis, Camp Responsive Element Modulator (CREM) pathway, and Androgen Receptor (AR) signaling crosstalk in MDD. Potential pathways in MDD associated with GPD include neurotransmission, cell differentiation, androgen signaling, and estrogen signaling. Fumarine, m-cresol, quercetin, betasitosterol, fumarine, taraxasterol, and lupeol in GPD may be the targets of SLC6A4, monoamine oxidase A (MAOA), DRD2, OPRM1, HTR3A, Albumin (ALB), and NTRK1, respectively. The IC50 values of trifolin targeting Phosphodiesterase (PDE) 4A and girinimbine targeting PDE4B1 were 73.79 μM and 31.86 μM, respectively. The IC50 values of girinimbine and benzo[a]carbazole on PDE4B2 were 51.62 μM and 94.61 μM, respectively. Conclusion: Different compounds in GPD may target the same protein, and the same component in GPD can target multiple targets. These results suggest that the effects of GPD on MDD are holistic and systematic, unlike the pattern of one drug-one target.

Aim: Hepatocellular carcinoma (HCC) is the world's second leading cause of cancerrelated mortality and the fifth most prevalent cancer overall. Several synthetic and plant-based remedies are in practice to treat diverse liver disorders. Because of their minimal side effects and protective characteristics, plant phenolics have the potential to become alternative therapeutics, replacing currently existing HCC medications. The present study identifies the plant phenolics as having the capacity to inhibit HCC with low side effects and cost efficiency. Background: Hepatocellular carcinoma (HCC) is the leading cause of cancer-related mortality, despite the proven effectiveness of screening programs for at-risk individuals, the majority of patients have disease progression or tumor characteristics that preclude curative therapies at the time of diagnosis. Acteoside (Verbascoside) is a naturally occurring phenylethanoid glycoside found throughout the plant kingdom. Acteoside is a physiologically active chemical with the number of pharmacological and protective effects against various liver illnesses. Objectives: Currently used HCC medications have a variety of side effects. Plant-based chemicals offer the possibility of treating HCC with minimal side effects. The work is targeted to find the best phytochemical (plant phenolic) lead molecule for future drug development research against Hepatocellular carcinoma. Methods: The targets were selected based on an analysis of relevant literature, and the 3D structures of the selected receptors were obtained in. pdb format from the RCSB-Protein data bank (PDB, http://www.rscb.org/pdb). Based on a review of the literature, sixty plant secondary metabolites, or plant phenolics, were selected. The ligand structures were obtained and downloaded in.sdf format from the NCBI PubChem chemicals database (https://pubchem.ncbi.nlm.nih.gov/). Molecular docking between the receptor and ligands was accomplished using the Molegro Virtual Docker 6.0 (MVD) software. Results: The target RAF1, BRAF chain 1, TIE2 chain 2 FGFR1, FGFR2, AXL, and FGFR4 showed the best binding effectiveness with acteoside compared to their respective positive control. RET chain 1 and BRAF chain 2 acteoside showed prominent binding efficacy after Curcumin, and Epigallocatechingallate, respectively, against positive control. Present findings clearly point towards the potentiality of acteoside in inhibiting various HCC targets. Conclusion: Acteoside may be used as a prominent lead molecule in the future treatment of hepatic cancer with its multifaceted binding efficiencies against various target proteins.

Background: Osteoporosis is a prevalent disease for the aged population. Chinese herb-derived natural compounds have anti-osteoporosis effects. Due to the complexity of chemical ingredients and natural products, it is necessary to develop a high-throughput approach with the integration of cheminformatics and deep-learning methods to explore their mechanistic action, especially herb/drug-gene interaction networks. Methods: Ten medicinal herbs for clinical osteoporosis treatment were selected. Chemical ingredients of top 10 herbs were retrieved from TCMIO database, and their predicted targets were obtained from SEA server. Anti-osteoporosis clinical drugs and targets were collected from multi-databases. Chemical space, fingerprint similarity, and scaffold comparison of the compounds between herbs and clinical drugs were analyzed by RDKit and SKlearn. A network of herb-ingredient-target were constructed via Gephi, and GO and KEGG enrichment analysis were performed using clusterProfiler. Additionally, the bioactivity of compounds and targets were predicted by DeepScreening. Molecular docking of YYH flavonoids to HSD17B2 was accomplished by AutoDockTools. Results: Cheminformatics result depicts a pharmacological network consisting of 89 active components and 30 potential genes. The chemical structures of plant steroids, flavonoids, and alkaloids are key components for anti-osteoporosis effects. Moreover, bioinformatics result demonstrates that the active components of herbs mainly participate in steroid hormone biosynthesis and the TNF signaling pathway. Finally, deep-learning-based regression models were constructed to evaluate 22 anti-osteoporosis-related protein targets and predict the activity of 1350 chemical ingredients of the 10 herbs. Conclusion: The combination of cheminformatics and deep-learning approaches sheds light on the exploration of medicinal herbs mechanisms, and the identification of novel and active compounds from medical herbs in complex molecular systems.

Aim: This study aimed to investigate how opioids affect phagocytosis and microglial nitrite and nitric oxide synthase (iNOS) production during inflammation. Background: Opioids are a group of chemicals that are naturally found in the opium poppy plant and exert a variety of effects on the brain, including pain alleviation in some cases. They are commonly used in surgery and perioperative analgesia. However, research on the impact of opioids on microglial inflammatory factor production and phagocytosis is limited. Objective: This study was designed to investigate the effects of opioids on inducible nitric oxide synthase (iNOS) activity and nitric oxide (NO) generation. Moreover, the influence of opioids on the engulfment of C8-B4 microglial cells after stimulation with LPS was also examined. Methods: C8-B4 mouse microglial cells were exposed to various concentrations of opioids after stimulation with lipopolysaccharide (LPS) and interferon-γ (IFN-γ). Nitrite production was assayed. The iNOS and Cox-2 were determined by Western blotting, and fluorescent immunostaining was performed to assess the percentage of microglia that engulfed fluorescent microspheres in total microglia cultivating with opioids after being activated by LPS. Results: After LPS and IFN-γ stimulation, microglia produced lower amounts of nitric oxide (NO) production with buprenorphine, salvinorin A, and naloxone (P Conclusion: Buprenorphine and salvinorin A were found to reduce NO production and iNOS induction in microglial cells activated by LPS and IFN-γ. Salvinorin A promoted the phagocytosis of microglia cells treated by LPS.

Background: Citrus grandis ‘Tomentosa,’ a fruit epicarp of C. grandis ‘Tomentosa’ or C. grandis (L.) Osbeck is widely used in health food and medicine. Based on our survey results, there are also rich essential oils with bioactivities in leaves, but the chemical compounds in this part and relevant pharmacological activities have never been studied systematically. Therefore, this study was to preliminarily decipher the pharmacological activities and mechanisms of the essential oil in leaves of C. grandis ‘Tomentosa’ by an integrated network pharmacology approach. Methods: Essential oil compositions from leaves of C. grandis ‘Tomentosa’ were identified using GC-MS/MS. And then, the targets of these oil compositions were predicted and screened from TCMSP, SwissTargetPrediction, STITCH and SEA databases. STRING database was used to construct the protein-protein interaction networks, and the eligible protein targets were input into WebGestalt 2019 to carry out GO enrichment and KEGG pathway enrichment analysis. Based on the potential targets, disease enrichment information was obtained by TTD databases. Cytoscape software was used to construct the component-target-disease network diagrams. Results: Finally, 61 essential oil chemical components were identified by GC-MS/MS, which correspond to 679 potential targets. Biological function analysis showed 12, 19, and 12 GO entries related to biological processes, cell components and molecular functions, respectively. 43 KEGG pathways were identified, of which the most significant categories were terpenoid backbone biosynthesis, TNF signaling pathway and leishmaniasis. The component-target-disease network diagram revealed that the essential oil compositions in leaves of C. grandis ‘Tomentosa’ could treat tumors, immune diseases, neurodegenerative diseases and respiratory diseases, which were highly related to CHRM1, PTGS2, CASP3, MAP2K1 and CDC25B. Conclusion: This study may provide new insight into C. grandis ‘Tomentosa’ or C. grandis (L.) Osbeck and may provide useful information for future utilization and development.

Drug–target interaction is crucial in the discovery of new drugs. Computational methods can be used to identify new drug–target interactions at low costs and with reasonable accuracy. Recent studies pay more attention to machine-learning methods, ranging from matrix factorization to deep learning, in the DTI prediction. Since the interaction matrix is often extremely sparse, DTI prediction performance is significantly decreased with matrix factorization-based methods. Therefore, some matrix factorization methods utilize side information to address both the sparsity issue of the interaction matrix and the cold-start issue. By combining matrix factorization and autoencoders, we propose a hybrid DTI prediction model that simultaneously learn the hidden factors of drugs and targets from their side information and interaction matrix. The proposed method is composed of two steps: the pre-processing of the interaction matrix, and the hybrid model. We leverage the similarity matrices of both drugs and targets to address the sparsity problem of the interaction matrix. The comparison of our approach against other algorithms on the same reference datasets has shown good results regarding area under receiver operating characteristic curve and the area under precision–recall curve. More specifically, experimental results achieve high accuracy on golden standard datasets (e.g., Nuclear Receptors, GPCRs, Ion Channels, and Enzymes) when performed with five repetitions of tenfold cross-validation. Graphical abstract: [Figure not available: see fulltext.]Display graphical of the hybrid model of Matrix Factorization with Denoising Autoencoderswith the help side information of drugs and targets for Prediction of Drug-Target Interactions.

Background: Sorafenib is the most widely used systematic therapy drug for treating unresectable Hepatocellular Carcinoma (HCC) but showed dissatisfactory efficacy in clinical applications. Objective: We conducted a combinational quantitative small-molecule high-throughput screening (qHTS) to identify potential candidates to enhance the treatment effectiveness of sorafenib. Methods: First, using a Hep3B human HCC cell line, 7051 approved drugs and bioactive compounds were screened, then the primary hits were tested with/without 0.5 μM sorafenib respectively, the compound has the half maximal Inhibitory Concentration (IC50) shift value greater than 1.5 was thought to have the synergistic effect with sorafenib. Furthermore, the MEK inhibitor PD198306 was selected for the further mechanistic study. Results: 12 effective compounds were identified, including kinase inhibitors targeting MEK, AURKB, CAMK, ROCK2, BRAF, PI3K, AKT and EGFR, and a μ-opioid receptor agonist and a Ltype calcium channel blocker. The mechanistic research of the combination of sorafenib plus PD198306 showed that the two compounds synergistically inhibited MEK-ERK and mTORC1- 4EBP1 and induced apoptosis in HCC cells, which can be attributed to the transcriptional and posttranslational regulation of MCL-1 and BIM. Conclusion: Small-molecule qHTS identifies MEK inhibitor PD1938306 as a potent sorafenib enhancer, together with several novel combination strategies that are valuable for further studies.

Background:: Pyroptosis is a novel form of programmed cell death in cancers, which regulates tumor cell invasion, proliferation, and metastasis, thereby affecting the prognosis of cancer patients. However, the role of Pyroptosis-Related Genes (PGs) in Hepatocellular Carcinoma (HCC) remains unclear. Methods:: Somatic mutation, copy number variation, and expression of 41 PGs were assessed in HCC and normal liver from the TCGA dataset. The Least Absolute Shrinkage and Selection Operator (LASSO) was used to construct the prognostic model. K-M curves, ROC curves, nomograph, and univariate and multivariate Cox regression were conducted to evaluate the predictive value of PGs. Immune infiltration was analyzed by CIBERSOFT and ssGSEA algorithm. The expression of prognostic PGs was validated by qPCR. Results:: Significant mutation and copy number variation of PGs were found in HCC. These genes were involved in an inflammatory response. In addition, 9 out of 41 PGs were differentially expressed in HCC and found to correlate significantly with patient survival. Then, these signature genes were selected to build a prognosis model and were utilized to stratify HCC patients into high and low PGs-score groups. It showed that the high-PGs group had a worse prognosis. Univariate and multivariate Cox regression verified that PGs-score was an independent risk factor for HCC. By ROC curves and nomogram, we showed that PGs-score effectively predicted the 1-, 3-, and 5-year survival of HCC patients and correlated with AFP level and disease stage. Immune infiltration analysis further showed that tumor immunity correlated with the PGs-score, and the expression of immune checkpoint molecule was significantly enhanced in the high PGs group. The PGs-score was also validated in the external validation cohort (ICGC). Finally, the expression of 9 signature genes was validated in normal liver and HCC cell lines. Conclusions:: This study elucidated the aberrant regulation of PGs in HCC, and those pyroptosisrelated genes may be applied as a prognostic factor of HCC.

Background: Nature has bestowed mother Earth with an array of herbals utilized as therapeutics for various human ailments since the origin of life. Bryonia laciniosa (family: Cucurbitaceae) is one such herb, which finds its mention in various traditional systems of medicine and has attracted current researchers due to its significant therapeutic value. Objective: The current article aims to present a literature metasynthesis on Bryonia laciniosa. Method: The authors performed scholarly searches for peer-reviewed findings on Bryonia laciniosa and incorporated all the data related to the phytochemical and therapeutic profile of the drug. Results: This compilation comprises of Phytochemical and Pharmacological profile of Bryonia laciniosa elaborating its traditional significance and recent researches related to its biological activities. The plant exhibits its potential as an antimicrobial, anti-inflammatory, analgesic, antipyretic, anticonvulsant, anti-asthmatic, anticancer, antioxidant, antidiabetic, and aphrodisiac agent. It also displays its benefits in wound healing and ulcerative colitis. Conclusion: The presence of flavonoids, saponins, terpenoids, anthocyanins, coumarins, alkaloids, polyphenols, tannins and emodins in this plant is responsible for its various pharmacological activities. The retrospective study provides direction for existing research as well as future studies to support the domain of pharmaceutical and medical sciences.

Aim: The aim of the study was to explore the efficacy as well as the mechanism of action of Pitongshu (PTS) on rats with functional dyspepsia (FD) induced by iodoacetamide gavage and tail clamping. Methods: The bioactive components of PTS were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), whereas the potential targets of PTS were obtained from the Similarity Ensemble Approach (SEA), TCMSP, and Swiss Target Prediction Database. The disease targets were obtained from the DisGeNET database, whereas Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using the R Software. The method of iodoacetamide gavage combined with tail clamping was used to establish the FD rat model in this study. Body weight, food intake, gastrointestinal motility, gastric acidity and secretion, and the mechanical pain threshold of rats were measured. The open-field test was also performed. The stomach and duodenum were histologically observed. The levels of serotonin (5-HT), Calcitonin Gene-Related Peptide (CGRP), Motilin (MTL), and Gastrin (GAS) in gastric tissues were detected by ELISA. Results: A total of 139 bioactive components and 17 potential targets of PTS were identified through a network pharmacology approach. The results of GO and KEGG enrichment analyses indicated that PTS could reduce the 5-HT secretion of gastric tissues through the serotonergic synaptic pathway and alleviate the symptoms of FD, indicating that PTS plays a therapeutic role. The results of animal experiments showed that PTS could increase body weight and food intake, improve autonomous activity, and decrease gastric acidity and secretion in FD rats. Furthermore, gastric sensitivity increased in FD rats, and PTS treatment could significantly decrease it. The results of ELISA showed that the overexpression of 5-HT and CGRP was decreased after PTS treatment in FD rats. Lastly, PTS could significantly improve gastrointestinal motility, as well as the levels of GAS and MTL in FD rats. Conclusion: PTS may reduce 5-HT secretion by regulating the serotonergic synaptic pathway, thereby reducing visceral sensitivity and alleviating the symptoms of FD.

Background: Liver Hepatocellular Carcinoma (LIHC) is characterized by high malignancy, poor prognosis, and high recurrence rate worldwide. The role of ferroptosis in tumorigenesis and progression has been confirmed in previous studies. However, the multi-omics analysis in liver cancer of ferroptosis-markers RPL8 remains to be elucidated. Methods: In this analysis, the RPL8 mRNA expression was analyzed via the GEPIA, TIMER and UALCAN databases. In addition, we verified the mRNA expression of RPL8 by qRT-PCR experiment. The Kaplan-Meier plotter, UALCAN, TCGAportal and HPA databases were applied to evaluate RPL8 on prognosis and clinicopathological parameters. Moreover, we used TIMER and Kaplan-Meier plotter to analyze the correlation of RPL8 to immune cell infiltration and immune cell type markers to prognosis. In addition, networks and function enrichment between RPL8 coexpression genes were analyzed by GeneMANIA, cBioportal and Metascape databases. What’s more, we used FerrDb and GEPIA databases to analyze the correlation of 23 Ferroptosis-related genes with RPL8. Results: The mRNA expression of RPL8 was over-expressed in multiple cancers. In addition, transcription and translation levels of RPL8 in LIHC were significantly higher than normal tissues. Furthermore, higher expression of RPL8 was closely related to shorter OS in LIHC patients. The analysis of Kaplan-Meier plotter proved that RPL8 expression was related to stage, Sorafenib treatment, alcohol consumption and hepatitis virus. Moreover, the results showed that the methylation expression level of RPL8 was significantly associated with age, gender, grade, stage and TP53 mutation of LIHC. RPL8 and its co-expression genes were primarily involved in liver regeneration and immune system process. Immune infiltration analysis showed the RPL8 expression had positively correlated with immune cells and immune subtypes in LIHC. Furthermore, qRT-PCR experiment validated the expression difference of RPL8 in liver cancer. Conclusion: Our findings elucidated that ferroptosis-markers RPL8 may play an important role in prognosis, and significantly correlate with ferroptosis-related genes, it also revealed the potential of RPL8 as a novel therapeutic target for LIHC treatment and prognosis assessment.

Background: Dental pulp stem cells (DPSCs) refer to a type of stem cells which is characterized by great differentiation potential and is easy to obtain. DPSCs are able to be employed for treating immune diseases and tissue regeneration. However, the differentiation ability exhibited by aging DPSCs is reduced, thereby limiting the application. As speculated by the microarray analysis, different expression of miRNAs might be involved in DPSC senescence, whereas comprehensive transcriptome level detection has been rare. Objective and Methods: To gain insights into the molecular mechanisms involved, RNA-seq, pathway enrichment and Gene Ontology Analysis were conducted on aging and young DPSCs. Results: In this study, the differences in lncRNA and mRNA expressions of the aging and young DPSCs were demonstrated, and the vital factors and the relevant pathways were speculated. On the whole, 18950 mRNAs and 21854 lncRNAs were detected, among which 14 mRNAs and 7 lncRNAs were differentially expressed. Furthermore, hsa-miR-6724-5p may be a vital node in the aging process of DPSCs, and its target genes was involved in the dopaminergic synapse. Conclusion: In brief, the aging of DPSCs was significantly dependent of different expressed genes (DEGs) which is related to dopaminergic synapse. However, the specific function and internal relationship of the DEGs should be verified in depth.

Angiotensin-converting enzyme 2 (ACE2) is a binding target of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein. An ACE2-like enzyme, such as bacterial M32-carboxypeptidase (M32-CAP), is assumed to be a potential therapeutic candidate for coronavirus disease 2019 (COVID-19). Here, we screened bacteria with an ACE2-like enzyme activity from Japanese fermented food and dietary products using the fluorogenic substrate for rapid screening. The strain showing the highest activity, Enterobacter sp. 200527-13, produced an enzyme with the same hydrolytic activity as ACE2 on Angiotensin II (Ang II). The enzymatic analysis using the heterologously-expressed enzyme in Escherichia coli revealed that the enzyme catalyzes the same reaction with that of ACE2, Ang II hydrolysis to Ang 1-7, and phenylalanine. The gene sequence information showed that the enzyme belongs to the M32-CAP family. These results suggested that the selected enzyme, M32-CAP (EntCP), from Enterobacter sp. 200527-13 was identified as an ACE2-like enzyme.

Background: Ningnanmycin is a new antibiotic pesticide with good bactericidal and antiviral efficacy, which is widely used in the control of fruit and vegetable diseases, and the excessive pesticide residues pose a serious threat to the environment and human health. Methods: In this study, we used fluorescence spectrometer to scan the three-dimensional spectrum of ningnanmycin samples. We used a BP neural network to complete the regression analysis of content prediction based on the fluorescence spectra. After that, the prediction performance of the BP neural network was compared with the exponential fitting method. Results: The results of the BP neural network modeling based on the obtained samples showed that the mean square error of the prediction results of the test set is less than 10-4, the R-square is greater than 0.99, the average recovery is 99.11%, and the model performance of the BP neural network is better than exponential fitting. Conclusion: Studies have shown that fluorescence spectroscopy combined with BP neural network can effectively predict the concentration of ningnanmycin.

Abstract: The present COVID-19 pandemic is terrible for the respiratory system and is caused by severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2). It has affected millions of people globally and over 511.9 million cases and 6.2 million deaths have been reported across the world. Various drugs have been repurposed, however, no specific medicine has been approved by the FDA to combat this disease till date. In this condition, researchers have been attracted to natural and safe products to improve immunity to viral infections through inhibiting viral cell entry, replication, and modulation. Various natural products, probiotics, and nutrients have antimicrobial, antiviral, analgesic, anti-inflammatory, and antiproliferative activities, and some of the compounds are also utilized in traditional medicine in Ayurveda, Siddha, and other cultures. This review provides a clinical perspective on the application of herbs for the prevention of viral infections.

Background: The lack of anti-COVID-19 treatment to date warrants urgent research into potential therapeutic targets. Virtual drug screening techniques enable the identification of novel compounds that target the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Main Protease (Mpro). Objective: The binding of the halogenated compounds to Mpro may inhibit the replication and transcription of SARS-CoV-2 and, ultimately, stop the viral life cycle. In times of dire need for anti- COVID-19 treatment, this study lays the groundwork for further experimental research to investigate these compounds' efficacy and potential medical uses to treat COVID-19. Method: New heterocyclic compounds were synthesized through the first reaction of cyclohexane- 1, 3-dione (1a) or dimedone (1b) with trichloroacetonitrile (2) to give the 2,2,2-trichloroethylidene) cyclohexane-1,3-dione derivatives 3a and 3b, respectively. The latter compounds underwent a series of heterocyclization reactions to produce biologically active compounds. Results: Novel compounds, including fused thiophene, pyrimidine and pyran derivatives, were synthesized and tested against human RNA N7-MTase (hRNMT) and selected viral N7-MTases such as SARS-CoV nsp14 and Vaccinia D1-D12 complex to evaluate their specificity and their molecular modeling was also studied in the aim of producing anti-covid-19 target molecules. Conclusion: The results showed that compounds 10a, 10b, 10c, 10e, 10f, 10g and 10h showed high % inhibitions against SARs-Covnsp 14. Whereas compounds 5a, 7a, 8b, 10a, 10b, 10c and 10i showed high inhibitions against hRNMT. This study explored the binding affinity of twenty-two halogenated compounds to the SARS-CoV-2 MPro and discovered fifteen compounds with higher binding affinity than Nelfinavir, of which three showed remarkable results. c-Met kinase inhibitions of 10a, 10f, 10g and 10h showed that all compounds exhibited higher inhibitions than the reference Foretinib.

Human dopamine transporter gene (SLC6A3) is one of the neurotransmission genes that plays an important role in controlling the behavior and psychological reactions toward various physical activities. The aim of the present study is to evaluate the associations between SLC6A3 3’-UTR VNTR (3’VNTR) genotypes and eight personality traits in Egyptian athletes. One hundred athletes (50 males and 50 females) and one hundred non-athletes participated in this investigation either in individual or team games. The Freiburg Personality inventory-Revised questionnaire was applied to athletes to evaluate personality traits according to the scale, at a scheduled time, using descriptive analysis. Genotyping for SLC6A3 3’-UTR VNTR polymorphism in athletes was realized to detect the different genotypes by PCR amplifications using desired primers. Three different genotypes of the 3’VNTR polymorphism; namely (9/9), (9/10), (10/10) were identified. The 9/10 and 10/10 genotypes of SLC6A3 gene were significantly higher in players than those in the control group. There were no statistically significant differences in personality traits values among males and females. The Cronbach’s alpha stability coefficient for four items is higher than 0.7 suggesting that these items (Neuroticism, Aggressiveness, Depression and Excitability) have a good reliability and support the used personality dimensions for this investigation. Our findings revealed a significant association between the genotypes of 3’VNTR and the personality traits in Egyptian athletes. These data shed light on the role of SLC6A3 gene in athletes’ behavioral performance and could serve as the basis for further characterization in the large number of Egyptian athletes.

This study examines the impact of Hofstede's six cultural dimensions, namely power distance, individualism/collectivism, masculinity/femininity, uncertainty avoidance, and long-term/short-term orientation on organizational behavior by comparing Indonesia with four different cultures: Vietnam, China, Taiwan, and France. It analyzes six scores in five different countries using a literature review on Hofstede's dimensions. With each dimension expressed on a scale of 0 to 100, In addition, this study translates the six dimensions into helping develop successful international business organizations. The results showed that Indonesia, China, Vietnam, Taiwan, and France all have a significant influence on organizational culture within the organization. In these countries, they show high levels of delegation of authority to employees, low levels of individualism and collectivism in China, Indonesia, Taiwan, and Vietnam, high levels of individualism in France, and masculinity and femininity, as well as uncertainty. On the long-orientation dimension, China and Taiwan score the highest among France, Indonesia, and Vietnam regarding indulgence versus restraint in combination with high scores on uncertainty avoidance, implying that Taiwanese and French are less relaxed and enjoy life less often than is usually assumed. that their actions are constrained by social norms. JEL classification numbers: D23, O19. Keywords: Hofstede’s dimension, National culture, Organizational culture, Organizational behavior.

This paper interrogates the concept of threshold concepts (TCs) of acids-bases in the teaching and learning of general chemistry of first-year university students. Acids-bases chemistry has been identified by chemistry scholars worldwide as one of the threshold concepts (TCs) in chemistry that seems to be difficult and the culprit of most failures in chemistry. Acids-bases reactions are also central to learning of other chemical reactions within the discipline of chemistry and other science concepts in life sciences (biology) and physics. The paper is based on the challenges students experience in general chemistry, which tend to discourage students from studying chemistry or pursuing their studies in chemistry-oriented professions, such as medicine, engineering, agriculture, natural resources, and chemistry education; or responsible for students dropping out of the university education completely. To address this problem, we explored lecturers’ and students’ challenges that students experience in learning TCs of acids-bases chemistry and the students’ perspectives of chemistry. A qualitative research methodology was employed in the study. Data was collected through classroom observations and student focus-group conversations. A thematic approach was employed to analyse data. Five chemistry educators and their classes were purposely sampled. Vygotsky’s social cognitive development theory was used as a theoretical framework to understand the teaching-learning of acids-bases chemistry. Findings suggested that some students’ experiences transformed them into self-directed learners. Researchers can investigate other avenues to the portal in teaching-learning TCs.

Although mathematical skills are an important component for business students, many students struggle to understand and meet the demands in different subjects. There is a considerable heterogeneity among the different subjects and the need for good mathematical skills varies from subject to subject. To analyse this, we apply the Attitudes Towards Mathematics Inventory (ATMI) framework. The data are collected by asking students at a business school in Norway. The results are linked to the exam grades in the various subjects. The sample consists of approximately 150 students. As a research method, the standard linear regression model is used. The results show that there is a strong positive correlation between ATMI and success in the various subjects. This applies specifically to the dimension of self-confidence in mathematics. However, there is a considerable variation between the subjects, and in some non-quantitative subjects such as marketing, no correlation was found in this study between ATMI and exam grades. As control variables in the model, gender and Grade Point Average (GPA) from the upper secondary school were included. These variables had a rather small influence on the results for the subjects, but with significant statistical effects on some subjects. This article provides insight into a key factor that can help explain how students are performing in the various business subjects

This paper critically discusses the everlasting but also extremely current, fundamental and of paramount significance issue concerning the training of secondary education teachers in pedagogy and didactics and focuses on the responsibility of the university professors who train the teachers at this school level. This topic is directly related to conducting the educational process. Consequently, the current education system of teachers at this school level is briefly presented which underlines their insufficient pedagogical and teaching training. Then, the several efforts that have been made to resolve the issue of teachers’ incompetence are outlined and analyzed, as well as the ascertainments and proposals that emerged from the attempts to resolve the issue are presented and discussed. Finally, the observations of the research study are discussed, and the corresponding conclusions are presented.

This paper explores the challenges associated with public procurement processes in the Czech Republic, as well as globally. In particular, it focuses on small-scale construction projects in the education sector and the problems associated with them, including policy making, pricing, efficiency, and the length of the procurement process. The paper also examines the risks associated with public procurement in the construction industry and how these risks can be managed through risk management. Moreover, it analyses the specific problems in the education sector, including imbalanced contractual conditions and unfair arrangements, which often shift the responsibility for risk aftermath to contractors. To gain insight into these issues, a research questionnaire was conducted among construction companies participating in the public procurement process. Based on the questionnaire results and a comparison between public and private procurement processes in the Czech Republic and Germany, the paper identifies flaws in the system and proposes potential solutions. The paper concludes by emphasizing the need for small changes to be implemented to improve the system, even though corruption remains a major issue that needs to be addressed. Overall, this paper provides valuable insights for policymakers and stakeholders on how to address the issues surrounding public procurement in the education sector in the Czech Republic and beyond.

Yu-Guo Yuan,1,2,* Ya-Xin Zhang,1,* Song-Zi Liu,1,* Abu Musa Md Talimur Reza,3 Jia-Lin Wang,1,2 Ling Li,1,2 He-Qing Cai,1,2 Ping Zhong,1 Il-Keun Kong4 1College of Veterinary Medicine, Yangzhou University, Yangzhou, People’s Republic of China; 2Jiangsu Co-Innovation Center of Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, People’s Republic of China; 3Department of Molecular Biology and Genetics Faculty of Basic Sciences, Gebze Technical University, Kocaeli, Republic of Turkiye; 4Division of Applied Life Science (BK21 Four), Institute of Agriculture and Life Science, Gyeongsang National University, Jinju, Gyeongnam Province, Republic of Korea*These authors contributed equally to this workCorrespondence: Yu-Guo Yuan; Il-Keun Kong, Email yyg9776430@163.com; ikong7900@gmail.comIntroduction: The increasing industrial and biomedical utilization of graphene oxide silver nanoparticles (GO-AgNPs) raises the concern of nanosafety: exposure to the AgNPs or GO-AgNPs increases the generation of reactive oxygen species (ROS), causes DNA damage and alters the expression of whole transcriptome including mRNA, miRNA, tRNA, lncRNA, circRNA and others. Although the roles of different RNAs in epigenetic toxicity are being studied during the last decade, but still we have little knowledge about the role of circle RNAs (circRNAs) in epigenetic toxicity.Methods: Rabbit fetal fibroblast cells (RFFCs) were treated with 0, 8, 16, 24, 32 and 48 μg/mL GO-AgNPs to test the cell viability and 24 μg/mL GO-AgNPs was selected as the experimental dose. After 24 h treatment with 24 μg/mL GO-AgNPs, the level of ROS, malondialdehyde (MDA), superoxide dismutase (SOD), intracellular ATP, glutathione peroxidase (GPx), and glutathione reductase (Gr) were measured in the RFFCs. High-throughput whole transcriptome sequencing was performed to compare the expression of circRNAs, long non-coding RNAs (lncRNA) and mRNA between 24 μg/mL GO-AgNPs-treated RFFCs and control cells. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to validate the accuracy of circRNA sequencing data. Bioinformatics analyses were performed to reveal the potential functional roles and related pathways of differentially expressed circRNAs, lncRNA and mRNA and to construct a circRNA–miRNA–mRNA interaction network.Results: We found that 57 circRNAs, 75 lncRNAs, and 444 mRNAs were upregulated while 35 circRNAs, 21 lncRNAs, and 186 mRNAs were downregulated. These differentially expressed genes are mainly involved in the transcriptional mis-regulation of cancer through several pathways: MAPK signaling pathway (circRNAs), non-homologous end-joining (lncRNAs), as well as PPAR and TGF-beta signaling pathways (mRNAs).Conclusion: These data revealed the potential roles of circRNAs in the GO-AgNPs induced toxicity through oxidative damage, which would be the basis for further research to determine their roles in the regulation of different biological processes.Keywords: circRNAs, GO-AgNPs, oxidative stress, epigenetic toxicity

Fengjun Xia,1,2,* Xiaoyan Tao,2,3,* Haichen Wang,1,2 Jian Shui,4 Changhang Min,1,2 Yubing Xia,1,2 Jun Li,1,2 Mengli Tang,1,2 ZhaoJun Liu,1,2 Yongmei Hu,1,2 Huidan Luo,1,2 Mingxiang Zou1,2 1National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 2Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 3Department of Laboratory Medicine and Sichuan Provincial Key Laboratory for Human Disease Gene Study, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, 610072, People’s Republic of China; 4Department of Clinical Laboratory, Changsha Central Hospital, Changsha, 410008, People’s Republic of China*These authors contributed equally to this workCorrespondence: Mingxiang Zou, National Clinical Research Center for Geriatric Disorders, Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, 410008, People’s Republic of China, Tel/Fax +86-7384327440, Email zoumingxiang@csu.edu.cnPurpose: As an under-explored biomaterial, bacterial biofilms have a wide range of applications in the green synthesis of nanomaterials. The biofilm supernatant of Pseudomonas aeruginosa PA75 was used to synthesize novel silver nanoparticles (AgNPs). BF75-AgNPs were found to possess several biological properties.Methods: In this study, we biosynthesized BF75-AgNPs using biofilm supernatant as the reducing agent, stabilizer, and dispersant and investigated their biopotential in terms of antibacterial, antibiofilm, and antitumor activities.Results: The synthesized BF75-AgNPs demonstrated a typical face-centered cubic crystal structure; they were well dispersed; and they were spherical with a size of 13.899 ± 4.036 nm. The average zeta potential of the BF75-AgNPs was − 31.0 ± 8.1 mV. The BF75-AgNPs exhibited strong antibacterial activities against the methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum beta-lactamase Escherichia coli (ESBL-EC), extensively drug-resistant Klebsiella pneumoniae (XDR-KP), and carbapenem-resistant Pseudomonas aeruginosa (CR-PA). Moreover, the BF75-AgNPs had a strong bactericidal effect on XDR-KP at 1/2 × MIC, and the expression level of reactive oxygen species (ROS) in bacteria was significantly increased. A synergistic effect was observed when the BF75-AgNPs and colistin were used for the co-treatment of two colistin-resistant XDR-KP strains, with fractional inhibitory concentration index (FICI) values of 0.281 and 0.187, respectively. Furthermore, the BF75-AgNPs demonstrated a strong biofilm inhibition activity and mature biofilm bactericidal activity against XDR-KP. The BF75-AgNPs also exhibited a strong antitumor activity against melanoma cells and low cytotoxicity against normal epidermal cells. In addition, the BF75-AgNPs increased the proportion of apoptotic cells in two melanoma cell lines, and the proportion of late apoptotic cells increased with BF75-AgNP concentration.Conclusion: This study suggests that BF75-AgNPs synthesized from biofilm supernatant have broad prospects for antibacterial, antibiofilm, and antitumor applications.Graphical Abstract: Keywords: multidrug resistant, green synthesis, colistin, metallic silver, melanoma

Background: Premature ovarian failure is a heterogeneous disease that severely affects the quality of life of women in their reproductive years. The ancient classical Chinese medicine compounds Zuo Gui Wan and You Gui Wan have great potential to treat premature ovarian failure, but the similarities and differences in their pharmacological mechanisms for treating POF are not yet clear. Method: In this study, the public database was used to screen the active ingredients and potential targets of Zuo Gui Wan and You Gui Wan. The similarities and differences in the potential targets of both pills for the treatment of POF were analysed using the POF-related genes obtained from OMIM and GeneCards. The protein–protein interaction network was established and collated to form a drug-active ingredient-target gene network by using STRING. Finally, the drug–target–pathway network was constructed by enrichment analysis to find the differences in target enrichment on the same pathway. Results: Pharmacological analysis of the network showed that Zuo Gui Wan contains 72 active ingredients, While You Gui Wan has 112. A total of 62 common compositions, such as quercetin and kaempferol, were identified. Amongst them were 10 unique compounds, such as hydroxyproline and cholesterol, in Zuo Gui Wan and 50 exclusive compounds, such as Karanjin and beta-carotene, in You Gui Wan. In addition, 14 overlapping targets, including MAPK1, CXCL8, TNF, IL6, and EGFR, were determined amongst the first 20 targets in the treatment of POF by both pills, demonstrating that the core mechanism of POF treatment is similar between the two. Pathway enrichment analysis showed 87 identical and significant pathways between Zuo Gui Wan and You Gui Wan, including IL-17, TNF, PI3K-Akt, oestrogen, VEGF, and other pathways. Zuo Gui Wan has 12 special pathways, such as natural killer cell-mediated cytotoxicity and intestinal immune network for IgA production. You Gui Wan has nine unique pathways, such as insulin secretion and glucagon signalling pathway. Conclusion: Zuo Gui Wan and You Gui Wan could treat POF by inhibiting oxidative stress and inflammation, regulating hormone levels, improving ovarian function, and promoting follicular development. Zuo Gui Wan is inclined to immune regulation, While You Gui Wan prefers insulin regulation. Therefore, similarities and differences clearly exist in the specific mechanisms of Zuo Gui Wan and You Gui Wan in the treatment of POF.

Background: P38α, emerging as a hot spot for drug discovery, is a member of the mitogen- activated protein kinase (MAPK) family and plays a crucial role in regulating the production of inflammatory mediators. However, despite a massive number of highly potent molecules being reported and several under clinical trials, no p38α inhibitor has been approved yet. There is still demand to discover novel p38α to deal with the safety issue induced by off-target effects. Objective: In this study, we performed a machine learning-based virtual screening to identify p38α inhibitors from a natural products library, expecting to find novel drug lead scaffolds. Method: Firstly, the training dataset was processed with similarity screening to fit the chemical space of the natural products library. Then, six classifiers were constructed by combing two sets of molecular features with three different machine learning algorithms. After model evaluation, the three best classifiers were used for virtual screening. Results: Among the 15 compounds selected for experimental validation, picrasidine S was identified as a p38α inhibitor with the IC50 as 34.14 μM. Molecular docking was performed to predict the interaction mode of picrasidine S and p38α, indicating a specific hydrogen bond with Met109. Conclusion: This work provides a protocol and example for machine learning-assisted discovery of p38α inhibitor from natural products, as well as a novel lead scaffold represented by picrasidine S for further optimization and investigation.

Li Lin,1,* Qingyong Chen,1,* Yue Dai,2 Yufeng Xia1 1Department of Pharmacognosy, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 211198, People’s Republic of China; 2Department of Traditional Chinese Medicine and Pharmacology, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 211198, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yufeng Xia, Department of Pharmacognosy, School of Traditional Chinese Pharmacy, China Pharmaceutical University, 639 Longmian Avenue, Jiangning District, Nanjing, 211198, People’s Republic of China, Tel +862583271400, Fax +862585301528, Email yfxiacpu@126.comPurpose: Madecassic acid (MCA) is a natural triterpenoid isolated from centellae herba that has diverse biological effects, such as anti-inflammatory, antioxidant, and anticancer activities. However, the efficacy of MCA is limited by low oral bioavailability caused by its extremely poor aqueous solubility. This study aimed to develop a self-nanoemulsifying drug delivery system (SNEDDS) for MCA to improve its oral absorption.Methods: The utilized oil phases, surfactants, and co-surfactants for SNEDDS were selected based on the solubility of MCA and emulsification efficiency. The optimized formulation was characterized for pharmaceutical properties and its pharmacokinetic behavior was examined in rats. Besides, the intestinal absorption property of MCA was investigated using in situ single-pass intestinal perfusion and intestinal lymphatic transport.Results: The optimized nanoemulsion formula consists of Capryol 90:Labrasol:Kolliphor ELP:Transcutol HP in a weight ratio of 1:2.7:2.7:3.6 (w/w/w/w). MCA-loaded SNEDDS presented a small droplet size (21.52 ± 0.23 nm), with a zeta potential value of − 3.05 ± 0.3 mV. Compared with pure MCA, SNEDDS had a higher effective permeability coefficient and showed 8.47-fold and 4.01-fold of maximum plasma concentration (Cmax) and area under the plasma concentration-time curve (AUC), respectively. Cycloheximide was pretreated before the experiment to evaluate the degree of lymphatic uptake. The results showed that cycloheximide greatly influenced the absorption of SNEDDS, resulting in 82.26% and 76.98% reduction in Cmax and AUC, respectively.Conclusion: This study reports the MCA-loaded SNEDDS with distinctly enhanced in vitro and in vivo performance compared with pure MCA and concludes that the SNEDDS formulation could be a viable and effective strategy for improving the dissolution rate and bioavailability of poor aqueous-soluble ingredients.Keywords: madecassic acid, self-nanoemulsifying drug delivery system, pharmacokinetics, intestinal lymphatic transport

Background: Cross-linked porous carbon nanofibers (CNF) was successfully prepared by electrospinning and high-temperature carbonization. Polyacrylonitrile (PAN) as the carbon source and Genipin as the cross-linking agent were used to prepare Cross-linked porous carbon nanofibers (CNF). Method and Material: The field emission Scanning Electron Microscopy (SEM), Transmission Electron Microscope (TEM), Automatic Specific Surface and Porosity Analyzer Brunner Emmet Teller (BET), X-ray Diffraction (XRD), A laser confocal microspectroscope (Raman, XploRA PLUS, Horiba) were used to characterize materials. The CNF suspension was dropped on the surface of the bare glassy carbon electrode by the drip coating method to obtain a CNF modified electrode. Cyclic voltammetry was used to study the electrochemiluminescence behavior of difenidol hydrochloride on CNF modified glassy carbon electrode (Glassy Carbon Electrode, GCE). Results and Discussion: Herein, we synthesised a kind of Crosslinked Carbon Nanofibers and designed a novel ECL biosensor, Under the optimal conditions, the concentration of difenidol hydrochloride has a linear relationship with the peak current in the range of 8.0×10-8 to 1.0×10-4mol/L, the correlation coefficient is R2=0.997, with a low detection limit (1.2×10- 8 mol/L). Difenidol hydrochloride in difenidol hydrochloride tablets was tested, and the recovery rate of sample addition was 83.17%-92.17%, and the RSD was

Haoyu Zhao,1,* Yachao Jia,1,* Feng Wang,1 Yimin Chai,1 Chunfu Zhang,2 Jia Xu,1 Qinglin Kang1 1Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Department of Orthopedic Surgery, Shanghai, People’s Republic of China; 2Shanghai Jiao Tong University, School of Biomedical Engineering, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jia Xu; Qinglin Kang, Email xujia0117@126.com; orthokang@163.comIntroduction: Large bone abnormalities are commonly treated using distraction osteogenesis (DO), but it is not suitable for a long-term application; therefore, there is an urgent need for adjuvant therapy that can accelerate bone repair.Methods: We have synthesized mesoporous silica-coated magnetic nanoparticles doped with cobalt ions (Co-MMSNs) and assessed their capacity to quicken bone regrowth in a mouse model of DO. Furthermore, local injection of the Co-MMSNs significantly accelerated bone healing in DO, as demonstrated by X-ray imaging, micro-CT, mechanical tests, histological evaluation, and immunochemical analysis.Results: In vitro, the Co-MMSNs exhibited good biocompatibility and induced angiogenic gene expression and osteogenic development in bone mesenchymal stem cells. And the Co-MMSNs can promote bone regeneration in a rat DO model.Discussion: This study demonstrated the significant potential of Co-MMSNs to shorten the DO treatment duration and effectively reduce the incidence of complications.Keywords: mesoporous silica-coated magnetic nanoparticles, mesenchymal stem cells, large bone defects, distraction osteogenesis, bone regeneration

Background: Experimental studies have shown that curcumin exerts neuroprotective effects in animal models with middle cerebral artery occlusion (MCAO). However, the mechanisms of protective effects of curcumin in MCAO are not fully understood. Objective: This study aims to investigate the key neurogenesis targets of curcumin action in mouse brain with MCAO. Methods: The MCAO models were established in mice. High-throughput sequencing was used to identify differentially expressed mRNA, lncRNA, and circRNA. The reverse expressed mRNAs, lncRNA, and circRNA in sham vs. MCAO and MCAO vs. curcumin were identified. Biological functions were determined by gene ontology (GO) analyses. The protein-protein interaction (PPI) network of neurogenesis-related genes was constructed. Next, neurogenesis-related lncRNA/ circRNA-miRNA-mRNA ceRNA networks were constructed. Results: The total of reverse expressed 1215 mRNAs, 32 lncRNAs, and 43 circRNAs were filtered based on the 2 series (sham vs. MCAO and MCAO vs. Curcumin). The functional enrichment analysis of 1215 reverse expressed mRNAs found that they were involved in neurogenesis, neuron generation, neurogenesis regulation, and others. The PPI network of neurogenesis-related genes consisted of 115 nodes, including 27 down-regulated genes and 36 up-regulated genes. Furthermore, the neurogenesis-related lncRNA/circRNA-miRNA-mRNA ceRNAs networks were constructed, and 5 lncRNA ceRNA networks and 3 circRNA ceRNA networks were explored. Conclusion: Our study revealed that curcumin exerts neuroprotective effects by regulating neurogenesis. The neurogenesis-related lncRNA/circRNA-miRNA-mRNA ceRNA networks are potential therapeutic targets of curcumin in MCAO. This study provided a theoretical basis for curcumin exerting neuroprotective effects in MCAO.

Jing Wang,1,* Weijian Chen,1,2,* Wenxiang Du,2 Hongjie Zhang,2 Matthias Ilmer,3 Lei Song,2 Yuan Hu,2 Xiaopeng Ma1 1Department of General Surgery, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, People’s Republic of China; 2State Key Laboratory of Fire Science, University of Science and Technology of China, Hefei, Anhui, 230026, People’s Republic of China; 3Department of General, Visceral, and Transplantation Surgery, Ludwig-Maximilians-University (LMU), Campus Grosshadern, Munich, 81377, Germany*These authors contributed equally to this workCorrespondence: Yuan Hu; Xiaopeng Ma, Email yuanhu@ustc.edu.cn; XiaopengMa@fsyy.ustc.edu.cnIntroduction: Tamoxifen (TAM) has proven to be a therapeutic breakthrough to reduce mortality and recurrence in estrogen receptor-positive (ER+) breast cancer patients. However, the application of TAM exhibits low bioavailability, off-target toxicity, instinct and acquired TAM resistance.Methods: We utilized black phosphorus (BP) as a drug carrier and sonosensitizer, integrated with TAM and tumor-targeting ligand folic acid (FA) to construct TAM@BP-FA for synergistic endocrine and sonodynamic therapy (SDT) of breast cancer. The exfoliated BP nanosheets were modified through in situ polymerization of dopamine, followed by electrostatic adsorption of TAM and FA. The anticancer effect of TAM@BP-FA was evaluated through in vitro cytotoxicity and in vivo antitumor model. RNA-sequencing (RNA-seq), quantitative real-time PCR, Western blot analysis, flow cytometry analysis and peripheral blood mononuclear cells (PBMCs) analysis were performed for mechanism investigation.Results: TAM@BP-FA had satisfactory drug loading capacity, the TAM release behavior can be controlled through pH microenvironment and ultrasonic stimulation. An amount of hydroxyl radical (∙OH) and singlet oxygen (1O2) were as expected generated under ultrasound stimulation. TAM@BP-FA nanoplatform showed excellent internalization in both TAM-sensitive MCF7 and TAM-resistant (TMR) cells. Using TMR cells, TAM@BP-FA displayed significantly enhanced antitumor ability in comparison with TAM (7.7% vs 69.6% viability at 5μg/mL), the additional SDT further caused 15% more cell death. RNA-seq unraveled the TAM@BP-FA antitumor mechanisms including effects on cell cycle, apoptosis and cell proliferation. Further analysis showed additional SDT successfully triggering reactive oxygen species (ROS) generation and mitochondrial membrane potential (MMP) reduction. Moreover, PBMCs exposed to TAM@BP-FA induced an antitumor immune response by natural killer (NK) cell upregulation and immunosuppression macrophage reduction.Conclusion: The novel BP-based strategy not only delivers TAM specifically to tumor cells but also exhibits satisfactory antitumor effects through targeted therapy, SDT, and immune cell modulation. The nanoplatform may provide a superior synergistic strategy for breast cancer therapy.Keywords: black phosphorus, breast cancer, sonodynamic therapy, tamoxifen, PBMC, combination therapy