Analysis of PANoptosis-related ceRNA network reveals lncRNA MIR17HG involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-α.

BACKGROUND: Inflammatory cytokines such as Interleukin 1ß(IL1ß), IL6,Tumor Necrosis Factor-α (TNF-α) can inhibit osteoblast differentiation and induce osteoblast apoptosis. PANoptosis, a newly identified type of programmed cell death (PCD), may be influenced by long noncoding RNA (lncRNAs) which play important roles in regulating inflammation. However, the potential role of lncRNAs in inflammation and PANoptosis during osteogenic differentiation remains unclear. This study aimed to investigate the regulatory functions of lncRNAs in inflammation and apoptosis during osteogenic differentiation. METHODS AND RESULTS: High-throughput sequencing was used to identify differentially expressed genes involved in osteoblast differentiation under inflammatory conditions. Two lncRNAs associated with inflammation and PANoptosis during osteogenic differentiation were identified from sequencing data and Gene Expression Omnibus (GEO) databases. Their functionalities were analyzed using diverse bioinformatics methodologies, resulting in the construction of the lncRNA-miRNA-mRNA network. Among these, lncRNA (MIR17HG) showed a high correlation with PANoptosis. Bibliometric methods were employed to collect literature data on PANoptosis, and its components were inferred. PCR and Western Blotting experiments confirmed that lncRNA MIR17HG is related to PANoptosis in osteoblasts during inflammation. CONCLUSIONS: Our data suggest that TNF-α-induced inhibition of osteogenic differentiation and PANoptosis in MC3T3-E1 osteoblasts is associated with MIR17HG. These findings highlight the critical role of MIR17HG in the interplay between inflammation, PANoptosis, and osteogenic differentiation, suggesting potential therapeutic targets for conditions involving impaired bone formation and inflammatory responses.

Investigation Driven by Network Pharmacology on Potential Components and Mechanism of DGS, a Natural Vasoprotective Combination, for the Phytotherapy of Coronary Artery Disease.

Phytotherapy offers obvious advantages in the intervention of Coronary Artery Disease (CAD), but it is difficult to clarify the working mechanisms of the medicinal materials it uses. DGS is a natural vasoprotective combination that was screened out in our previous research, yet its potential components and mechanisms are unknown. Therefore, in this study, HPLC-MS and network pharmacology were employed to identify the active components and key signaling pathways of DGS. Transgenic zebrafish and HUVECs cell assays were used to evaluate the effectiveness of DGS. A total of 37 potentially active compounds were identified that interacted with 112 potential targets of CAD. Furthermore, PI3K-Akt, MAPK, relaxin, VEGF, and other signal pathways were determined to be the most promising DGS-mediated pathways. NO kit, ELISA, and Western blot results showed that DGS significantly promoted NO and VEGFA secretion via the upregulation of VEGFR2 expression and the phosphorylation of Akt, Erk1/2, and eNOS to cause angiogenesis and vasodilation. The result of dynamics molecular docking indicated that Salvianolic acid C may be a key active component of DGS in the treatment of CAD. In conclusion, this study has shed light on the network molecular mechanism of DGS for the intervention of CAD using a network pharmacology-driven strategy for the first time to aid in the intervention of CAD.

Reduction of psychological cravings and anxiety in women compulsorily isolated for detoxification using autonomous sensory meridian response (ASMR).

OBJECTIVE: To explore the effects of the autonomous sensory meridian response (ASMR) on the psychological cravings and anxiety of women compulsorily isolated for detoxification. METHOD: Around 122 women were recruited in a female drug detoxification center. Except for the 12-week training of ASMR, the experimental conditions of the experimental group (n = 60) were the same as those of the control group (n = 62). The addiction Stroop task was used to assess the level of psychological cravings and the State-Trait Anxiety Inventory was used to assess the level of anxiety. RESULTS: After the training, the decrease in state anxiety of the experimental group was larger than that of the control group, and the reaction time of the experimental group in the Stroop was also significantly lower than before the training. CONCLUSIONS: ASMR could thus reduce to a certain extent the state anxiety and attentional bias for drug-related clues under signaling psychological cravings among women compulsorily isolated for detoxification. HIGHLIGHTS: Intervention effects on psychological cravings and anxiety of women isolated for detoxification Basis for role of ASMR in regulating psychological cravings and anxiety in forced abstainers ASMR intervention reduced forced abstainers' attentional bias to drug-related clues.

[Effect of dual sustained-release nanofiber scaffolds embedded with BMP-2 protein nanospheres on the proliferation and differentiation of MC3T3-E1 cells].

PURPOSE: Electrospinning technology was used to construct PCL composite nanofiber scaffold material of BMP-2 sustained-release nanospheres, and the effect of this nanospheres on proliferation and differentiation of MC3T3-E1 cells was evaluated. METHODS: Solvent removal method and electrostatic self-assembly technology were used to prepare BMP-2 loaded chitosan nanospheres, the morphology, particle size and composition, BMP-2 protein encapsulation efficiency, and in vitro sustained release were tested. Electrospinning technology was ued to prepare PCL composite scaffold material containing BMP-2 nanospheres, and its morphology, hydrophilicity,and sustained release of BMP-2 protein were examined. In vitro cytology experiment was conducted to observe the growth of cells in the material, and the formation of ALP, related genes, and mineralized nodules during the process of osteogenic differentiation of the material were detected. SPSS 21.0 software package was used for statistical analysis. RESULTS: The nanospheres structure with stable structure was successfully prepared, with a high drug loading rate and sustained release of BMP-2. The PCL/BNPs scaffold material group had good hydrophilic properties and was conducive to cell proliferation and differentiation. The results of in vitro cell experiments showed that the cells spread well on the scaffold and the number of adhesions increased. ALP and related osteogenic genes COL1, OPN, RUNX2 increased, and the size and number of calcium nodules increased significantly. CONCLUSIONS: The polycaprolactone composite fiber scaffold material of BMP-2 sustained-release nanospheres can provide a new choice for the development of bone tissue engineering.

Exploring the binding mechanism of HDAC8 selective inhibitors: Lessons from the modification of Cap group.

The abnormal expression of histone deacetylase 8 (HDAC8) has been reported to associate with various cancer entities (colon, breast cancer, pancreas, etc.) as well as parasitic diseases, making HDAC8 gradually develop into an attractive and potential therapeutic target. Among the various design strategies of selective HDAC8 inhibitors (modification of Cap, Linker, or zinc binding group regions), the optimization of Cap region has aroused great interest among the researchers. However, the detailed information underlying how the modification of Cap region influences the inhibitory activities is still unclear, and in this study, compounds 2c, 3g, and 3n were selected to explore the differences in binding mechanisms brought by Cap modifications via various computational approaches at the atomic level. Five residues (Y293, H167, D254, D165, and M261) have a large difference in energy contributions to the constructed systems, and the subpocket formed by Y293 and M261 could interact with Cap groups, triggering the differences in the energy contributions of the residues (H167, D254, and D165) located in metal-catalytic center. In summary, the compounds 2c, 3g, and 3n were selected as molecular probes to explore the binding mechanism, and the residues (Y293 and M261) forming the subpocket should be paid special attention in the design and synthesis of novel selective HDAC8 inhibitors.

[Effect of concentrated growth factor on the biological properties of MC3T3-E1 osteoblasts].

PURPOSE: To observe the effect of concentrated growth factor(CGF) on the biological properties of osteoblasts. METHODS: MC3T3-E1 cells were cultured in CGF environment and a blank control group was established. The adhesion of osteoblasts to CGF surface was observed by scanning electron microscopy. Cell proliferation and alkaline phosphatase(ALP) activity were detected at 1, 4 and 7 d by Cell Counting Kit-8 (CCK-8) and Alkaline Phosphatase Assay Kit. The expression of mineralized nodules and osteogenesis-related gene Runx2 was observed by alizarin red staining. CGF extract was cultured for 24 h. Peptide staining was used to observe morphological changes in the cytoskeleton. SPSS 21.0 software package was used for statistical analysis. RESULTS: CCK-8 showed cells incubated for 1, 4 and 7 d in the experimental group had a stronger proliferation ability compared with the control group, and the difference was statistically significant (P<0.05). ALP activity test showed that there was no significant difference between the experimental group and the control group (P>0.05) at 1 d; but after 4 days of culture, cell in the experimental group had an increased ALP activity compared with the control group, and the difference was statistically significant(P<0.05). The results of alizarin red staining showed that the number of calcified nodules in the CGF group increased and the area was larger. In the phalloidin staining and DAPI staining, the number of cells in the CGF group increased, the cell spreading surface increased, and the actin shape was clearer. CGF significantly promoted Runx2 mRNA expression(P<0.05). CONCLUSIONS: High concentration of CGF can promote the proliferation and differentiation of MC3T3-E1 cells and the expression level of related osteogenic gene Runx2.

Online comprehensive two-dimensional hydrophilic interaction chromatography×reversed-phase liquid chromatography coupled with hybrid linear ion trap Orbitrap mass spectrometry for the analysis of phenolic acids in Salvia miltiorrhiza.

In the present study, a system was developed for the analysis of phenolic acids in Salvia miltiorrhiza using online comprehensive two-dimensional hydrophilic interaction chromatography and reversed-phase liquid chromatography coupled to a DAD detector and hybrid linear ion trap-Orbitrap mass spectrometry (HILIC×RP-DAD-ESI/HRMS/MSn). The system was configured based on the combination of an XBridge Amide column (150mm×4.6mm, 3.5µm) and Accucore PFP column (50mm×4.6mm, 2.6µm) for the first and second dimensions, respectively. An additional LC pump was used to dilute the eluent from the first dimension to decrease its elution strength in the second dimension. A back-flush trap column was selected as an interface to make up for the loss of efficiency and resolution due to the online dilution. Under the optimized conditions, a total of 196 peaks of polar compounds were successfully separated and detected in Salvia miltiorrhiza using the developed online HILIC×RP system, which exhibited high orthogonality (73%). The online combination of HILIC and RP provides powerful separation capability for the analysis of polar compounds in samples with complex matrices.

Main active constituent identification in Guanxinjing capsule, a traditional Chinese medicine, for the treatment of coronary heart disease complicated with depression.

Guanxinjing capsules (GXJCs) are used in traditional Chinese medicine as a common therapy for coronary heart disease (CHD) complicated with depression. In this study, we aimed to identify the main active constituents in GXJCs and to investigate the mechanisms of GXJC action on CHD complicated with depression. The chemical constituent profile of the GXJC was identified by UHPLC-LTQ-Orbitrap assay, and oral bioavailability was evaluated to screen the GXJC drug-like chemical constituents. A total of 16 GXJC drug-like chemical constituents were identified. Then, putative targets of the GXJC drug-like chemical constituents were predicted using MedChem Studio, with 870 genes found to be the putative targets of these molecules. After that, a GXJC putative target-known CHD/depression therapeutic target network was constructed, and four topological features, including degree, betweenness, closeness and K-coreness, were calculated. According to the topological feature values of the GXJC putative targets, 14 main active constituents were identified because their corresponding putative targets had topological importance in the GXJC putative target-known CHD/depression therapeutic target network, which were defined as the candidate targets of GXJC against CHD complicated with depression. Functionally, these candidate targets were significantly involved in several CHD/depression-related pathways, including repairing pathological vascular changes, reducing platelet aggregation and inflammation, and affecting patient depression. This study identified a list of main active constituents of GXJC acting on CHD complicated with depression using an integrative pharmacology-based approach that combined active chemical constituent identification, drug target prediction and network analysis. This method may offer an efficient way to understand the pharmacological mechanisms of traditional Chinese medicine prescriptions.

Hepatorenal protective effects of medicinal herbs in An-Gong-Niu-Huang Wan (AGNH) against cinnabar- and realgar-induced oxidative stress and inflammatory damage in mice.

An-Gong-Niu-Huang Wan (AGNH) is a famous traditional Chinese medicine prescription that contains cinnabar (HgS) and realgar (As2S2); the clinical practice of AGNH is hindered because both mercury and arsenic are hepatorenal toxic metalloids. It is noted that the cinnabar and realgar in AGNH are not used alone, but rather combined with different kinds of medicinal herbs as a formula to use. In this study, we evaluated the hepatorenal protective effects of the medicinal herbs in AGNH after co-exposure to cinnabar and realgar for 4 weeks in mice. The combination of the herbs in AGNH alleviated cinnabar and realgar-induced histopathological alterations and oxidative stress in the liver and kidneys. Furthermore, in cinnabar and realgar-treated mice, the increased expression levels of inducible enzymes (COX-2 and iNOS) and proinflammatory mediators (IL-1ß, TNF-α, PGE2 and NO) in the liver and kidneys were consistently down-regulated when medicinal herbs were combined as a formula. We also found that the herbs could reduce the inflammatory response by the inactivation of the MAPK and PI3K/Akt signaling pathway and the resulting blockade of NF-κB activation. Overall, our data indicates that the herbal medicines in AGNH attenuate cinnabar and realgar-induced hepatorenal toxicity by improving antioxidant competence and suppressing inflammatory injury.

[Fragmentation pathways of five furocoumarine using line ion trap with orbitrapmass spectrometry].

OBJECTIVE: The multiple levels fragmentations of five furocoumarine (psoralen, xanthotoxin, bergapten, oxypeucedanin, and byakangelicol) in Angelica dahurica have been demonstrated using LTQ-Orbitrap mass spectrometry with high resolution and high mass accuracy to discover the possible,fragmentation regularity. METHOD: Duringcollsion-induced dissociation (CID), the MS(n) data of the five compoundswhich were gained in the positive ion mode at 35ev collision energy by direct injection syrings method were analyzed using Xcalibar 2.0 Software to infer the formula of these fragmentations. RESULT: The results indicated that the five compounds have similar fragmentation process with CO meutral lost at C5,C8-subsituents and furan ring, meanwhile the meutralloss of CO2 occurred easily at lactone group. CONCLUSION: This method is helpful in identifying the structures of other furocoumarinein Angelica dahuricaand their metabolites in vivo.

[Rapid identification of chemical composition in safflower with UHPLC-LTQ-Orbitrap].

The UHPLC-LTQ-Orbitrap high resolution mass spectrometer was used to explore the chemical compositions in safflower. The rapid separation of the compositions was conducted by the UHPLC, following by high resolution full scan and MS2 scan, under the positive and negative ion mode. The chemical formula of compositions were deduced by full scan data in less than 5, then the potential structures were confirmed by the MS2 data. Forty-nine compounds were detected, of which 26 was identified, and 5 compounds was validated by the standard substances.

Deciphering the pharmacological mechanism of the Chinese formula huanglian-jie-du decoction in the treatment of ischemic stroke using a systems biology-based strategy.

AIM: Huanglian-jie-du decoction (HLJDD) is an important multiherb remedy in TCM, which is recently demonstrated to be effective to treat ischemic stroke. Here, we aimed to investigate the pharmacological mechanisms of HLJDD in the treatment of ischemic stroke using systems biology approaches. METHODS: Putative targets of HLJDD were predicted using MetaDrug. An interaction network of putative HLJDD targets and known therapeutic targets for the treatment of ischemic stroke was then constructed, and candidate HLJDD targets were identified by calculating topological features, including 'Degree', 'Node-betweenness', 'Closeness', and 'K-coreness'. The binding efficiencies of the candidate HLJDD targets with the corresponding compositive compounds were further validated by a molecular docking simulation. RESULTS: A total of 809 putative targets were obtained for 168 compositive compounds in HLJDD. Additionally, 39 putative targets were common to all four herbs of HLJDD. Next, 49 major nodes were identified as candidate HLJDD targets due to their network topological importance. The enrichment analysis based on the Gene Ontology (GO) annotation system and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway demonstrated that candidate HLJDD targets were more frequently involved in G-protein-coupled receptor signaling pathways, neuroactive ligand-receptor interactions and gap junctions, which all played important roles in the progression of ischemic stroke. Finally, the molecular docking simulation showed that 170 pairs of chemical components and candidate HLJDD targets had strong binding efficiencies. CONCLUSION: This study has developed for the first time a comprehensive systems approach integrating drug target prediction, network analysis and molecular docking simulation to reveal the relationships between the herbs contained in HLJDD and their putative targets and ischemic stroke-related pathways.

[Study on action mechanism of adjuvant therapeutic effect compound Ejiao slurry in treating cancers based on network pharmacology].

Recently, compound Ejiao slurry (FFEJJ) had been applied to treat cancer patients in clinic, with obvious curative effect. In this study, data and literatures were collected from the TCM chemical component database to establish the chemical component database of FFEJJ. Afterwards, MetaDrug software was used to predict the targets of FFEJJ and obtain the compound-target network. Next, the compound-target network was compared and analyzed to obtain the "compound-target-tumor target" heterogeneous network. Besides, further analysis was made on gene functions and metabolic pathway. The results indicated that FFEJJ could directly resist tumors by regulating cancer cell differentiation, growth, proliferation and apoptosis, and show an adjuvant therapeutic effect by enriching the blood and increasing the immunity.

Health literacy issues in the care of Chinese American immigrants with diabetes: a qualitative study.

OBJECTIVES: To investigate why first-generation Chinese immigrants with diabetes have difficulty obtaining, processing and understanding diabetes related information despite the existence of translated materials and translators. DESIGN: This qualitative study employed purposive sampling. Six focus groups and two individual interviews were conducted. Each group discussion lasted approximately 90 min and was guided by semistructured and open-ended questions. SETTING: Data were collected in two community health centres and one elderly retirement village in Los Angeles, California. PARTICIPANTS: 29 Chinese immigrants aged ≥45 years and diagnosed with type 2 diabetes for at least 1 year. RESULTS: Eight key themes were found to potentially affect Chinese immigrants' capacity to obtain, communicate, process and understand diabetes related health information and consequently alter their decision making in self-care. Among the themes, three major categories emerged: cultural factors, structural barriers, and personal barriers. CONCLUSIONS: Findings highlight the importance of cultural sensitivity when working with first-generation Chinese immigrants with diabetes. Implications for health professionals, local community centres and other potential service providers are discussed.

[Study on absorption location of four components from Naoxintong capsule].

OBJECTIVE: To study the absorption characteristics of four components from Naoxintong capsule in intestines. METHOD: In vitro everted gut sac method was adopted for preparing the intestinal absorption solution of Naoxintong capsule. UPLC was used to detect the content of chemical components in different intestinal segments, and comparing the results with the absorption of chemical components of Naoxintong capsule in each intestinal segment. The time-accumulative absorption curve was drawn to observe the changes in the accumulative absorption concentration with time. RESULT: Four ingredients of Naoxintong capsule can be detected in intestinal absorption solution, they are ferulic acid, paeoniflorin, salvianolic acid B and hydroxysafflor yellow A. Specifically, the accumulative absorption concentrations of ferulic acid, salvianolic acid B and paeoniflorin in ileum and rear jejunum segments were higher than that in front and middle jejunum segments; the absorption of ferulic acid, paeoniflorin, salvianolic acid B and hydroxysafflor yellow A did not reach saturated conditions in 3 hours. CONCLUSION: Ferulic acid, paeoniflorin, salvianolic acid B and hydroxysafflor yellow A are absorbed in the whole intestine. Ferulic acid, paeoniflorin and salvianolic acid B may be absorbed in specific segments.