HnRNP A2/B1 as a potential anti-tumor target for triptolide based on a simplified thermal proteome profiling method using XGBoost.

BACKGROUND: Triptolide (TP) is a highly active natural medicinal ingredient with significant potential in anticancer. The strong cytotoxicity of this compound suggests that it may have a wide range of targets within cells. However, further target screening is required at this stage. Traditional drug target screening methods can be significantly optimized using artificial intelligence (AI). PURPOSE:  This study aimed to identify the direct protein targets and explain the multitarget action mechanism of the anti-tumor effect of TP with the help of AI. METHODS:  The CCK8, scratch test, and flow cytometry analysis were used to examine cell proliferation, migration, cell cycle, and apoptosis in tumor cells treated with TP in vitro. The anti-tumor effect of TP in vivo was evaluated by constructing a tumor model in nude mice. Furthermore, we established a simplified thermal proteome analysis (TPP) method based on XGBoost (X-TPP) to rapidly screen the direct targets of TP. RESULTS: We validated the effects of TP on protein targets through RNA immunoprecipitation and pathways by qPCR and Western blotting. TP significantly inhibited tumor cell proliferation and migration and promoted apoptosis in vitro. Continuous administration of TP to tumor mice can significantly suppress tumor tissue size. We verified that TP can affect the thermal stability of HnRNP A2/B1 and exert anti-tumor effects by inhibiting HnRNP A2/B1-PI3K-AKT pathway. Adding siRNA to silence HnRNP A2/B1 also significantly down-regulated expression of AKT and PI3K. CONCLUSION: The X-TPP method was used to show that TP regulates tumor cell activity through its potential interaction with HnRNP A2/B1.

Advanced data post-processing method for rapid identification and classification of the major triterpenoids of Alismatis rhizoma by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry.

INTRODUCTION: Alismatis rhizoma (AR), a distinguished diuretic traditional Chinese herbal medicine, is widely used for the treatment of diarrhea, edema, nephropathy, hyperlipidemia, and tumors in clinical settings. Most beneficial effects of AR are attributed to the major triterpenoids, whose contents are relatively high in AR. To date, only 25 triterpenoids in AR have been characterized by LC-MS because the low-mass diagnostic ions are hardly triggered in MS, impeding structural identification. Herein, we developed an advanced data post-processing method with abundant characteristic fragments (CFs) and neutral losses (NLs) for rapid identification and classification of the major triterpenoids in AR by UPLC-Q-TOF-MSE . OBJECTIVE: We aimed to establish a systematic method for rapid identification and classification of the major triterpenoids of AR. METHODS: UPLC-Q-TOF-MSE coupled with an advanced data post-processing method was established to characterize the major triterpenoids of AR. The abundant CFs and NLs of different types of triterpenoids were discovered and systematically summarized. The rapid identification and classification of the major triterpenoids of AR were realized by processing the data and comparing with information described in the literature. RESULTS: In this study, a total of 44 triterpenoids were identified from AR, including three potentially new compounds and 41 known ones, which were classified into six types. CONCLUSION: The newly established approach is suitable for the chemical profiling of the major triterpenoids in AR, which could provide useful information about chemical constituents and a basis for further exploration of its active ingredients in vivo.

Chemical and metabolic profiling of Codonopsis Radix extract with an integrated strategy using ultra-high-performance liquid chromatography coupled with mass spectrometry.

Codonopsis radix was commonly used as food materials or herbal medicines in many countries. However, the comprehensive analysis of chemical constituents, and in vivo xenobiotics of Codonopsis radix remain unclear. In the present study, an integrated strategy with feature-based molecular networking using ultra-high-performance liquid chromatography coupled with mass spectrometry was established to systematically screen the chemical constituents and the in vivo xenobiotics of Codonopsis radix. A step-by-step manner based on a composition database, visual structure classification, discriminant ions, and metabolite software prediction was proposed to overcome the complexities due to the similar structure of chemical constituents and metabolites of Codonopsis radix. As a result, 103 compounds were tentatively characterized, 20 of which were identified by reference standards. Besides, a total of 50 xenobiotics were detected in vivo, including 26 prototypes and 24 metabolites, while the metabolic features of the pyrrolidine alkaloids were elucidated for the first time. The metabolism reactions of pyrrolidine alkaloids and sesquiterpene lactones included oxidation, methylation, hydration, hydrogenation, demethylation, glucuronidation, and sulfation. This study provided a generally applicable approach to the comprehensive investigation of the chemical and metabolic profile of traditional Chinese medicine and offered reasonable guidelines for further screening of quality control indicators and pharmacodynamics mechanism of Codonopsis radix.

Simultaneous determination of multiple constituents of Qi-Lin pill by UPLC-MS/MS: Applications to pharmacokinetics and testicular tissue distribution in rats.

Qi-Lin pill (QLP) is an effective traditional Chinese medicine prescription (TCMP) that has been used for the treatment of the oligoasthenozoospermia in China. Recently, some articles described the pharmacological effects of QLP and multiple ingredients in QLP contribute to its effects. However, the pharmacokinetic and target tissue distribution data of QLP are still unknown. In the present study, according to the Bioanalytical Method Validation Guidance of FDA, a sensitive and selective UPLC-MS/MS method was developed and validated for simultaneous determination of multiple constituents in rat plasma and testicular tissue, including morusimic acid A, codonopyrridium B, magnoflorine, emodin, 2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glucoside (THSG), ecliptasaponin A, paeoniflorin, albiflorin, gallic acid, danshensu, salvianolic acid A, catechin, isosinensetin, nobiletin, formononetin, calycosin, icariside II, icariin and epimedin C. For 19 analytes, the LLOQs reached 0.01-4 ng/mL. And all calibration curves showed favorable linearity (r ≥ 0.9903) in linear ranges. The intra-day and inter-day precision (relative standard deviation) for all analytes was less than 14.92 %, and the accuracies (as relative error) were in the range of - 6.44 % to 6.22 %. Extraction recoveries and matrix effects of analytes and IS were acceptable. All analytes were stable during the assay and storage in plasma samples. The method was successfully applied for the pharmacokinetics and testis distribution of multiple chemical constituents in QLP after a single oral dose. As a result, high exposure of danshensu, gallic acid, paeoniflorin and albiflorin were observed in rat plasma and testicular tissue. Among the flavonoids, isosinensetin and nobiletin had high exposure in testicular tissue. Moreover, alleviation of progesterone reduction was evaluated in H2O2-induced R2C leydig cells, and danshensu, gallic acid, paeoniflorin, albiflorin and nobiletin showed potent activity. Therefore, these five components were considered to be the effective components of QLP due to their relatively high exposure in vivo and biological activity. This finding also provided relevant information on action mechanism of QLP in the treatment of oligoasthenozoospermia.

Targeted isolation and identification of bioactive pyrrolidine alkaloids from Codonopsis pilosula using characteristic fragmentation-assisted mass spectral networking.

Codonopsis pilosula (CP), a well-known food medicine homology plant, is commonly used in many countries. In our preliminary study, a series of pyrrolidine alkaloids with high MS responses were detected as characteristic absorbed constituents in rat plasma after oral administration of CP extract. However, their structures were unclear due to the presence of various isomers and the lack of reference standards. In the present study, an MS-guided targeted isolation of pyrrolidine alkaloids of CP extract was performed by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC/Q-TOF MS). For data analysis under fast data directed acquisition mode (Fast-DDA), an effective approach named characteristic fragmentation-assisted mass spectral networking was successfully applied to discover new pyrrolidine alkaloids with high MS response in CP extract. As a result, seven new pyrrolizidine alkaloids [codonopyrrolidiums C-I (3-9)], together with two known ones (1 and 2), were isolated and identified by NMR spectral analysis. Among them, codonopyrrolidium B (1), codonopyrrolidium D (4) and codonopyrrolidium E (5) were evaluated for lipid-lowering activity, and they could improve high fructose-induced lipid accumulation in HepG2 cells. In addition, the characteristic MS/MS fragmentation patterns of these pyrrolizidine alkaloids were investigated, and 17 pyrrolidine alkaloids were identified. This approach could accelerate novel natural products discovery and characterize a class of natural products with MS/MS fragmentation patterns from similar chemical scaffolds. The research also provides a chemical basis for revealingin vivo effective substances in CP.

Prognostic role of the prognostic nutritional index in patients with pancreatic cancer who underwent curative resection without preoperative neoadjuvant treatment: A systematic review and meta-analysis.

Background: The prognostic nutrition index (PNI), which has been evaluated in various kinds of cancers, offered a simple yet effective approach to predict the prognosis. The aim of this meta-analysis is to reveal the correlation between preoperative PNI and the prognosis of patients with pancreatic ductal adenocarcinoma (PDAC) who underwent curative resection. Methods: We searched the PubMed, Embase, Web of Science and Cochrane Library databases, and extracted the hazard ratio (HR) with 95% confidential interval (CI) from eligible studies. The pooled HR with 95% CI was applied to evaluate the association between PNI and overall survival (OS), recurrence-free survival (RFS). Results: A total of fourteen studies with 3,385 patients were included for meta-analysis. The results (the pooled HR: 1.664, 95% CI: 1.424-1.994, I² = 42.6%, p value = 0.046) indicated that low preoperative PNI was closely related to poor OS. In addition, the results suggested that PNI was negatively correlated with RFS (the pooled HR: 1.369, 95%CI: 1.080-1.734). The robustness of these pooled results was verified by our subgroup analysis and sensitivity analysis. Moreover, different cutoff values among studies are responsible for the heterogeneity of pooled HR of OS through meta-regression analysis (p value = 0.042). Funnel plots, Begg's test (p value = 0.228) and Egger's test (p value = 0.702) indicated no significant publication bias in OS. Conclusion: Preoperative PNI might be a promising marker to predict the prognosis of PDAC patients who underwent curative resection.

Determination of luteolin in Chrysanthemum tea with a ultra-sensitive electrochemical sensor based on MoO3/poly(3,4-ethylene dioxythiophene)/gama-cyclodextrin metal-organic framework composites.

Chrysanthemum tea is a tranditional Chinese health drink, which contains luteolin, a flavonoid with vesatile health benefit activities. Herein, A sensitive electrochemical sensor based on composite materials consisting of MoO3 nanorods, poly (3, 4-ethylene dioxyethiophene)(PEDOT), and γ-cyclodextrin metal-organic framework(CD-MOF) was prepared.The materials were characterized and analyzed by scanning electron microscopy (SEM), X-ray diffraction (XRD), fourier transform infrared (FTIR), and X-ray photoelectron spectroscopy (XPS). Due to the synergisticeffects of the materials, the sensor showed a wide linear range of 0.4 nM -1800 nM and a low detection limit (LOD) of 0.1 nM (S/N = 3) for luteolin under optimized conditions. Besides, the influences of some coexistent phenolic compounds and common metal ions on luteolin detection were evaluated and no significant interference was observed. Finally, the sensor was successfully applied to the detection of luteolin in real Chrysanthemum tea samples.

Targets preliminary screening for the fresh natural drug molecule based on Cosine-correlation and similarity-comparison of local network.

BACKGROUND: Chinese herbal medicine is made up of hundreds of natural drug molecules and has played a major role in traditional Chinese medicine (TCM) for several thousand years. Therefore, it is of great significance to study the target of natural drug molecules for exploring the mechanism of treating diseases with TCM. However, it is very difficult to determine the targets of a fresh natural drug molecule due to the complexity of the interaction between drug molecules and targets. Compared with traditional biological experiments, the computational method has the advantages of less time and low cost for targets screening, but it remains many great challenges, especially for the molecules without social ties. METHODS: This study proposed a novel method based on the Cosine-correlation and Similarity-comparison of Local Network (CSLN) to perform the preliminary screening of targets for the fresh natural drug molecules and assign weights to them through a trained parameter. RESULTS: The performance of CSLN is superior to the popular drug-target-interaction (DTI) prediction model GRGMF on the gold standard data in the condition that is drug molecules are the objects for training and testing. Moreover, CSLN showed excellent ability in checking the targets screening performance for a fresh-natural-drug-molecule (scenario simulation) on the TCMSP (13 positive samples in top20), meanwhile, Western-Blot also further verified the accuracy of CSLN. CONCLUSIONS: In summary, the results suggest that CSLN can be used as an alternative strategy for screening targets of fresh natural drug molecules.

[Molecular mechanism of Fagopyri Dibotryis Rhizoma in treatment of acute lung injury based on network pharmacology and in vitro experiments].

The present study explored the mechanism of Fagopyri Dibotryis Rhizoma(FDR) and its main active components in the treatment of acute lung injury(ALI) based on the network pharmacology and the in vitro experiments. The main active components of FDR were obtained from the TCMSP database and screened by oral bioavailability and drug-likeness. The related target proteins of FDR were retrieved from the PubChem database, and the target genes related to ALI were screened out from the GeneCards database. A protein-protein interaction(PPI) network of compound target proteins and ALI target genes was constructed using STRING 11.0. Ingenuity Pathway Analysis(IPA) platform was used to analyze the common pathways of the potential compound target proteins of FDR and ALI target genes, thereby predicting the key targets and potential signaling pathways of FDR for the treatment of ALI. Finally, the potential pathways and key targets were verified by the in vitro experiments of lipopolysaccharide-induced RAW264.7 cells intervened by epicatechin(EC), the active component of FDR. The results of network pharmacology showed that 15 potential active components such as EC, procyanidin B1, and luteolin presumedly functioned in the treatment of ALI through nuclear transcription factor-κB(NF-κB) signaling pathway, transforming growth factor-ß(TGF-ß) signaling pathway, and adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK) signaling pathway through key targets, such as RELA(P65). The results of in vitro experiments showed that 25 µmol·L~(-1) EC had no toxicity to cells and could inhibit the expression of the p65-phosphorylated protein in the NF-κB signaling pathway to down-regulate the expression of downstream inflammatory cytokines, including tumor necrosis factor-α(TNF-α), IL-1ß and nitric oxide(NO), and up-regulate the expression of IL-10. These results suggested that the therapeutic efficacy of FDR on ALI was achieved by inhibiting the phosphorylation of p65 protein in the NF-κB signaling pathway and down-regulating the level of proinflammatory cytokines downstream of the signaling pathways.

Hyperbaric oxygen via mediating SIRT1-induced deacetylation of HMGB1 improved cReperfusion inj/reperfusion injury.

Ischemic stroke leads to severe neurological dysfunction in adults. Hyperbaric oxygen (HBO) induces tolerance to cReperfusion inj/reperfusion (I/R) injury. Therefore, our aims were to investigate whether SIRT1 participates in regulatingin the neuro-protective effect of HBO in a cerebral I/R model and its mechanism. Mice N2a cells were used to construct an oxygen deprivation/reperfusion (OGD/R) model to simulate in vitro brain I/R injury and to evaluate the role of HBO in OGD/R stimulated cells. Cell proliferation was detected using MTT, and apoptosis was determined by flow cytometry. ELISA was used to measure the concentration of TNF-α, IL-1ß and IL-6 related inflammatory factors. RT-qPCR and western blot assays were performed to test the expression of SIRT1. Immunoprecipitation was used to detect acetylation of HMGB1. Expression of SIRT1 was obviously reduced after OGD/R treatment in N2a cells, while SIRT1 was obviously enhanced in HBO treated cells. Moreover, knockdown of SIRT1 induced neuro-inflammation damage in cells and HBO effectively improved the inflammatory response in OGD/R treated cells by affecting SIRT1 levels. Furthermore, HBO induced the deacetylation of HMGB1 via regulating SIRT1. Interestingly, HBO via regulating the SIRT1-induced HMGB1 deacetylation and suppressing MMP-9 improved ischemic brain injury. HBO regulated ischemic brain injury via regulation of SIRT1-induced HMGB1 deacetylation, making it a potential treatment for ischemic brain injury treatment.

Adjuvant treatment with Angong Niuhuang pills in treating traumatic brain damage: a meta-analysis of randomized controlled trials.

BACKGROUND: The Angong Niuhuang pill (ANP) has been widely used in the adjuvant treatment of patients with traumatic brain injury (TBI). However, the efficacy and adverse reactions of this drug are controversial. In this study, it was aimed to evaluate the effectiveness and safety of ANP on patients with TBI by a systematic review and meta-analysis of randomized controlled trials (RCTs). METHODS: PubMed, Embase, Cochrane Library, Chinese Biomedicine Database (CBM), China National Knowledge Infrastructure (CNKI), Chinese Scientific Journals Database (VIP), and Wangfang databases were systematically searched from their establishment until June 2020. RCTs of ANP treating TBI were enrolled. Odds risk (OR) was used to assess the total effective rate and safety and mean difference (MD) and 95% confidence interval (CI) were used to assess the quantitative data. Tthe included literature's quality was evaluated by RevMan 5.3. The sensitivity and publication bias was evaluated by Stata 16.0. RESULTS: Twelve studies were identified in this systematic review, including 1,568 participants. The metaanalysis results suggested that ANP combined with routine treatment obviously improved the postoperative GCS [MD =1.97, 95% CI (1.22, 2.72), P<0.01] and GOS [OR =2.28, 95% CI (1.60, 3.22), P<0.01] of patients with TBI. ANP also increased Mg2+ concentration and decreased pulmonary infection. In addition, ANP significantly reduced NSE, gastrointestinal bleeding, and liver and kidney function damage. CONCLUSIONS: Based on limited evidence, ANP adjuvant therapy may have a clinical benefit in improving the prognosis of patients with TBI and reducing the associated complications. At the same time, more studies with larger sample sizes and high quality are required to determine the safety and effectiveness of ANP adjuvant therapy.

Facilitating effects of plant hormones on biomass production and nutrients removal by Tetraselmis cordiformis for advanced sewage treatment and its mechanism.

Advanced sewage treatment by microalgae is regarded as a promising method for addressing eutrophication. To improve sewage treatment, three kinds of plant hormones including auxin (indole-3-acetic acid, IAA), cytokinin (Zeatin), and brassinosteroid, were chosen to measure the influence of plant hormones on nitrogen and phosphorus removal by Tetraselmis cordiformis and to analyze their mechanisms, including photosynthesis, nutrient metabolism, and gene transcription. The results indicated that the maximal removal efficiencies of total nitrogen and phosphate by T. cordiformis were elevated by the plant hormones by 184.3% and 53.2%, respectively. The chlorophyll a content was increased by 1.1 times by the plant hormones in comparison with the control. Moreover, after being stimulated by plant hormones, the activities of nitrate reductase (NR) and glutamine synthetase (GS) increased by 90.4% and 82.1%, respectively, in comparison with the control. Supplementation with plant hormones also significantly elevated the mRNA expression level of GS-related gene by 30.9%. This study demonstrated that plant hormones could significantly promote the nutrient removal of microalgae for sewage treatment in artificial laboratory conditions and provided theoretical support for its further practical full-scale application under variable conditions.

Establishment and efficiency analysis of a single-stage denitrifying phosphorus removal system treating secondary effluent.

For advanced phosphorus and nitrogen removal, denitrifying phosphorus removal (DPR) was used to treat secondary effluent of sewage plants based on alternating anoxic/anaerobic process within single-stage biofilter. Under the hydraulic load of 3 m3/(m2·h), average removal rates of TP and TN in the system were 61.05% and 90.54%. 82.7% of the NO3--N removal occurred in the upper of the packing layer. TP removal occurred in upper and lower of the packing layer, accounting for 42.02% and 57.98% of the total removal, respectively. Biomass and bioactivity decreased proportional to the height incensement of packing layer. Nitrogen and phosphorus removal rates increased with anaerobic time while decreased with hydraulic load. 16S rDNA sequencing results showed dominant DNPAOs in the system included Acinetobacter and Dechloromonas, while dominant denitrifying bacteria included Flavobacterium, Comamonadaceae, Hydrogenophaga, Thauera and Azospira. The study further provided an effective and feasible way for advanced wastewater treatment.

[Preventive effects of jinghua weikang capsule on NSAID-induced injury to the mucosa of the small intestine: an experimental research].

OBJECTIVE: To study the preventive effects of jinghua weikang capsule (JWC) on nonsteroidal anti-inflammatory drugs (NSAIDs) induced injury to the mucosa of the small intestine. METHODS: Thirty-two Wistar rats were randomly divided into four groups, i.e., the blank control group, the model group, the JWC group, and the esomeprazole group. Diclofenac was administered to rats in the model group, the JWC group, and the esomeprazole group at the daily dose of 15 mg/kg. JWC and esomeprazole was respectively given to those in the JWC group, and the esomeprazole group one day ahead. Normal saline was given to rats in the blank control group. Rats were killed 3 days later. The pathological changes of the small intestine were observed by hematoxylin and eosin stain. RESULTS: Compared with the blank control group, the general score for the small intestine (4.63 +/-0.52 vs 0.00 +/-0. 00) and the pathological score (4.00 +/-0.90 vs 0.00 +/-0. 00) obviously increased in the model group, showing statistical difference (P <0.05). Compared with the model group, the general score for the small intestine (1.88 +/-0.99) and the pathological score (2.11 +/-1.11) obviously decreased in the JWG group, showing statistical difference (P <0.05). Compared with the model group, the general score for the small intestine (2.75 +/-1.28) and the pathological score (2. 30 +/-0.94) obviously decreased in the esomeprazole group, showing statistical difference (P <0.05). CONCLUSION: JWC could prevent NSAIDs induced injury to the mucosa of the small intestine.

A proteomical study on the radiosensitized target molecules of fuzheng zengxiao formula in pulmonary adenocarcinoma nude mice model.

OBJECTIVE: To investigate the radiosensitized target of Fuzheng Zengxiao Formula. METHODS: The pulmonary adenocarcinoma (PAa) nude mice of tumor transplantation model were prepared and divided into four groups: Group I (blank control group, n = 10), Group II (simple radiotherapy group, n = 10), Group III (radiotherapy plus Fuzheng Zengxiao Formula, n = 10) and Group IV (radiotherapy plus metronidazole, n = 10). Radiation of X-rays was given to the tumors in Group I, II and III when they were averagely about 1 centimetre in diameter. 23 hours later, the tumors were taken, the total proteins were extracted, and the protein contents were determined. The proteins were isolated with two dimensional gel electrophoresis, and the differentially expressed proteins were analyzed with mass spectrometry and identified by protein database. RESULTS: Six significant proteins, including apolipoprotein E, ceratin75, S100A9, cyclophilin A, S100A10 and hemoglobin, were determined. Compared with Group I, apolipoprotein E and ceratin75 highly expressed in the Group II; compared with Group II, S100A9, cyclophilin A and hemoglobin had high expression in the Group III; compared with Group II, S100A9, cyclophilin A, S100A10 and hemoglobin had high expression in the Group IV; compared with Group IV, S100A9 and S100A10 had low expression and hemoglobin had high expression in Group III. CONCLUSION: The radiosensitization of Fuzheng Zengxiao Formula is related with the improvement of hypoxia state; and possibly S100A9 and cyclophilin A are the target proteins of Fuzheng Zengxiao Formula in radiosensitization.