Glucocorticoids ameliorate cardiorenal syndrome through the NPR1/SGK1 pathway in natriuretic peptide receptor A­heterozygous mice.

Natriuretic peptides, which are produced by the heart, bind to natriuretic peptide receptor A (NPR1 encoded by natriuretic peptide receptor 1 gene) and cause vasodilation and natriuresis. Thus, they serve an important role in regulating blood pressure. In the present study, microinjection of CRISPR associated protein 9/single guide RNA into fertilized C57BL/6N mouse eggs was performed to generate filial generation zero (F0) Npr1 knockout homozygous mice (Npr1-/-). F0 mice mated with wild-type (WT) mice to obtain F1 Npr1 knockout heterozygous mice with stable heredity (Npr1+/-). F1 self-hybridization was used to expand the population of heterozygous mice (Npr1+/-). The present study performed echocardiography to investigate the impact of NPR1 gene knockdown on cardiac function. Compared with those in the WT group (C57BL/6N male mice), the left ventricular ejection fraction, myocardial contractility and renal sodium and potassium excretion and creatinine-clearance rates were decreased, indicating that Npr1 knockdown induced cardiac and renal dysfunction. In addition, expression of serum glucocorticoid-regulated kinase 1 (SGK1) increased significantly compared with that in WT mice. However, glucocorticoids (dexamethasone) upregulated NPR1 and inhibited SGK1 and alleviated cardiac and renal dysfunction caused by Npr1 gene heterozygosity. SGK1 inhibitor GSK650394 ameliorate cardiorenal syndrome by suppressing SGK1. Briefly, glucocorticoids inhibited SGK1 by upregulating NPR1, thereby ameliorating cardiorenal impairment caused by Npr1 gene heterozygosity. The present findings provided novel insight into the understanding of cardiorenal syndrome and suggested that glucocorticoids targeting the NPR1/SGK1 pathway may be a potential therapeutic target to treat cardiorenal syndrome.

Hydrogel dressing integrating FAK inhibition and ROS scavenging for mechano-chemical treatment of atopic dermatitis.

Atopic dermatitis (AD) is a chronic skin disease caused by skin immune dyshomeostasis and accompanied by severe pruritus. Although oxidative stress and mechanical scratching can aggravate AD inflammation, treatment targeting scratching is often overlooked, and the efficiency of mechano-chemically synergistic therapy remains unclear. Here, we find that enhanced phosphorylation of focal adhesion kinase (FAK) is associated with scratch-exacerbated AD. We then develop a multifunctional hydrogel dressing that integrates oxidative stress modulation with FAK inhibition to synergistically treat AD. We show that the adhesive, self-healing and antimicrobial hydrogel is suitable for the unique scratching and bacterial environment of AD skin. We demonstrate that it can scavenge intracellular reactive oxygen species and reduce mechanically induced intercellular junction deficiency and inflammation. Furthermore, in mouse AD models with controlled scratching, we find that the hydrogel alleviates AD symptoms, rebuilds the skin barrier, and inhibits inflammation. These results suggest that the hydrogel integrating reactive oxygen species scavenging and FAK inhibition could serve as a promising skin dressing for synergistic AD treatment.

The Mechanism of Dynamic Interaction between Doxorubicin and Calf Thymus DNA at the Single-Molecule Level Based on Confocal Raman Spectroscopy.

It is of great fundamental significance and practical application to understand the binding sites and dynamic process of the interaction between doxorubicin (DOX) and DNA molecules. Based on the Confocal Raman spectroscopy, the interaction between DOX and calf thymus DNA has been systemically investigated, and some meaningful findings have been found. DOX molecules can not only interact with all four bases of DNA molecules, i.e., adenine, thymine, cytosine, guanine, and phosphate, but also affect the DNA conformation. Meanwhile, the binding site of DOX and its derivatives such as daunorubicin and epirubicin is certain. Furthermore, the interaction between DOX and DNA molecules is a dynamic process since the intensities of each characteristic peaks of the base, e.g., adenine, cytosine, and phosphate, are all regularly changed with the interaction time. Finally, a dynamic mechanism model of the interaction between DOX and DNA molecules is proposed; that is, there are two kinds of interaction between DOX and DNA molecules: DOX-DNA acts to form a complex, and DOX-DOX acts to form a multimer. The two effects are competitive, as the former compresses DNA molecules, and the latter decompresses these DNA molecules. This work is helpful for accurately understanding and developing new drugs and pathways to improve and treat DOX-induced cytotoxicity and cardiotoxicity.

Rational Construction of a Ni/CoMoO4 Heterostructure with Strong Ni-O-Co Bonds for Improving Multifunctional Nanozyme Activity.

Due to the lack of a general descriptor to predict the activity of nanomaterials, the current exploration of nanozymes mainly depended on trial-and-error strategies, which hindered the effective design of nanozymes. Here, with the help of a large number of Ni-O-Co bonds at the interface of heterostructures, a prediction descriptor was successfully determined to reveal the double enzyme-like activity mechanisms for Ni/CoMoO4. Additionally, DFT calculations revealed that interface engineering could accelerate the catalytic kinetics of the enzyme-like activity. Ni-O-Co bonds were the main active sites for enzyme-like activity. Finally, the colorimetric signal and intelligent biosensor of Ni/CoMoO4 based on deep learning were used to detect organophosphorus and ziram sensitively. Meanwhile, the in situ FTIR results uncovered the detection mechanism: the target molecules could block Ni-O-Co active sites at the heterostructure interface leading to the signal peak decreasing. This study not only provided a well design strategy for the further development of nanozymes or other advanced catalysts, but it also designed a multifunctional intelligent biosensor platform. Furthermore, it also provided preferable ideas regarding the catalytic mechanism and detection mechanism of heterostructure nanozymes.

Insight into the role of dermal white adipose tissue loss in dermal fibrosis.

The loss of dermal white adipose tissue (dWAT) is vital to the formation of dermal fibrosis (DF), but the specific mechanism is not well understood. A few studies are reviewed to explore the role of dWAT in the formation of DF. Recent findings indicated that the adipocytes-to-myofibroblasts transition in dWAT reflects the direct contribution to the DF formation. While adipose-derived stem cells (ADSCs) contained in dWAT express antifibrotic cytokines, the loss of ADSCs leads to skin protection decreased, which indirectly exacerbates DF and tissue damage. Therefore, blocking or reversing the adipocytes-to-myofibroblasts transition or improving the survival of ADSCs in dWAT and the expression of antifibrotic cytokines may be an effective strategy for the treatment of DF.

The m6A RNA Modification Modulates Gene Expression and Fibrosis-Related Pathways in Hypertrophic Scar.

Purpose: To systematically analyze the overall m6A modification pattern in hyperplastic scars (HS). Methods: The m6A modification patterns in HS and normal skin (NS) tissues were described by m6A sequencing and RNA sequencing, and subsequently bioinformatics analysis was performed. The m6A-related RNA was immunoprecipitated and verified by real-time quantitative PCR. Results: The appearance of 14,791 new m6A peaks in the HS sample was accompanied by the disappearance of 7,835 peaks. The unique m6A-related genes in HS were thus associated with fibrosis-related pathways. We identified the differentially expressed mRNA transcripts in HS samples with hyper-methylated or hypo-methylated m6A peaks. Conclusion: This study is the first to map the m6A transcriptome of human HS, which may help clarify the possible mechanism of m6A-mediated gene expression regulation.

A Stable NanoPAA-ZnO/ZnCl2 Composite with Variable 3D Structured Morphology and Sustained Superhydrophilicity.

A ZnO/ZnCl2 composite with stable 3D structural morphologies and long lasting superhydrophilicity was synthesized on the top surface of a nano porous anodic alumina (nanoPAA) substrate. The wettability of a nanoPAA-ZnO/ZnCl2 was systematically characterized and the experimental data indicated that the water contact angle (WCA) of 0° could be achieved as well as maintained over 7 days and still remained at 4.36° after 50 days, and its 3D structural morphology had no clearly observable change during this period. The mechanism for the superhydrophilicity of the composites was interpreted in terms of the inherent hydrophilicity of ZnO/ZnCl2 nanofilm, the three-dimensional structures of wrinkled nanoflakes, the nanogaps between neighbor nanoflakes, the difference of structual morphologies (i.e., size, shape, and upright posture of nanoflakes), and the measured True Volume of voids in the nanocomposite. The structural morphologies were mainly determined by the parameters such as the original concentration of precursor ZnCl2 and the pore diameter of nanoPAA substrate. The study proposes a promising superhydrophilic nanomaterial and a cost-effective synthesis method, which will play a practical role in the fields of biomedical molecular sensors and micro/nanofluidic chips.

Hollow MnFeO oxide derived from MOF@MOF with multiple enzyme-like activities for multifunction colorimetric assay of biomolecules and Hg2.

In this work, a new type of hollow MnFeO oxide derived obtained from the metal-organic framework (MOF)@MOF was designed and proposed, which has intrinsic activity of mimicking enzymes of oxidase and peroxidase by adjustment pH values. Based on the colorimetric reaction and the hindrance of the oxidase-like activity in the presence of L-cysteine (Cys), as well as the recovery of oxidase-like activity due to the specific complexation of Cys and mercury (II) ions (Hg2+), a new type of colorimetric transmission platform for Cys and Hg2+ detection with wide linear ranges of 1-25 µM for Cys and 0.1-15 µM for Hg2+ has been developed. Besides, a better colorimetric sensing platform for detecting H2O2 was established with linear ranges of 1-60 µM and 60-300 µM based on generating hydroxyl radicals (·OH). Furthermore, the hollow MnFeO oxide has high stability, excellent selectivity with good activity over a long period of time. Surprisingly, the proposed method for Cys, Hg2+ and H2O2 estimation can also be used in actual samples. These characteristics lay a foundation for further investigation about the catalytic activity of the hollow MnFeO oxide nanomaterials and make it show broad prospects in the field of biosensing and catalysis.

Exploring the effect of C-X-C motif chemokine ligand-13 on the proliferation and migration of mesenchymal stem cells based on network pharmacology / 解剖学报

Objective To explore the effect of C-X-C motif chemokine ligand-13 (CXCL-13) on the proliferation and migration of human bone marrow mesenchymal stem cells (BMSCs) by network pharmacology. Methods To predict that the targets of CXCL-13 on BMSCs by online database. Metascape was used to perform gene ontology (GO) of the targets and Kyoto encyclopedia of genes and genomes (KEGG) pathway was used to perform enrichment analysis. The protein interaction analysis was performed by STRING 11.0 database, and the protein module of core gene was screened by using the cytoHubba 0. 1 of Cytoscape 3. 8. We divided BMSCs into control group, CXCL-13 group and PI3K inhibitor group. MTT assay, flow cytometric analysis and Transwell cell migration assay were respectively used to detect the absorbance (A) value of BMSCs in each group, the apoptosis rate and the number of cell migration. The protein contents of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) in BMSCs supernatant were determined by ELISA. Western blotting was used to detect the protein expression of Akt and phosphorylated Akt (p-Akt) of BMSCs in each group. Results It was predicted that 21 targets of CXCL-13 effect on BMSCs. There were 32 biological processes related to cell proliferation include stem cell proliferation, regulation of endothelial cell proliferation and positive regulation of smooth muscle cell proliferation. There were 22 biological processes related to cell migration include regulating cell migration, amebic cell migration and endothelial cell migration. There were 40 KEGG pathways including cancer pathway, PI3K-Akt signaling pathway and MAPK signaling pathway. The core proteins included tumor protein P53 (TP53), epidermal growth factor receptor (EGFR), heat shock protein 90 kD alpha class B member 1 (HSP90AB1), protein kinase Ca (PRKCA), estrogen receptor 2 (ESR2) and prostaglandin E receptor 4 (PTGER4). Compared with other groups, the absorbance (A) value and cell migration number of BMSCs in CXCL-13 group increased significantly (P< 0. 01, 71=15), and the apoptosis rate decreased significantly (P<0. 01, n= 15). However, absorbance value, apoptosis rate and migration number of BMSCs in PI3K inhibitor group were contrary to those in CXCL-13 group (P<0. 01, n= 15). Compared with the control group, the protein contents of EGF and VEGF in BMSCs of CXCL-13 group increased significantly (P<0. 01, n= 15), and the relative expression of Akt and p-Akt increased significantly (P<0. 01, n = 9). However, the protein content of EGF and VEGF, and the relative expression of Akt and p-Akt in PI3K inhibitor group were opposite. Conclusion Through activating PI3K-Akt pathway, CXCL-13 may promote BMSCs paracrine EGF and VEGF proteins, and improve proliferation and migration of BMSCs, as well as inhibit BMSCs apoptosis.

[Product improvement and secondary development of Pudilan Xiaoyan Oral Liquid based on clinical medication].

Pudilan Xiaoyan Oral Liquid is widely used in clinical applications, with safe and effective results. Its coverage rate in the national first, second and third grade hospitals is as high as 71%. In this study, we analyzed and summarized the research progress on the material basis, quality control, production process and clinical medication of Pudilan Xiaoyan Oral Liquid based on the clinical diseases(parotitis, tonsillitis, pharyngitis), and deeply explored the intrinsic quality improvement and secondary development of Pudilan product. Pharmacodynamic material basis of Pudilan Xiaoyan Oral Liquid was explored through the network pharmacology technology and quality control indicators of the production process were optimized by cell anti-inflammatory experiments. Through these techno-logies, it would be more specific, scientific and effective to carry out process optimization of each link and multidimensional quality control of the whole process. The dosage and oral compliance for special patients(children) were explored, providing a reference for clinical pediatric medication of Pudilan Xiaoyan Oral Liquid. Simultaneously, it is helpful to expand the application market by developing Pudilan daily chemical products, and promote the traditional Chinese medicine products in terms of curative effect and daily life.

[Investigation of active components and mechanism of Pudilan Xiaoyan Oral Liquid based on network pharmacology].

To investigate the active components/ingredients of Pudilan Xiaoyan Oral Liquid based on the network pharmacology technology, and analyze the network data of its potential targets and mechanisms. The active ingredient screening, protein interaction analysis and pathway annotation were used to further optimize its active components and potential targets, and clarify the pharmacodynamic substance basis and mechanism of Pudilan Xiaoyan Oral Liquid. Through this technique, we screened out 41 active ingredients in Pudilan Xiaoyan Oral Liquid, mainly including 16 alkaloid components, 13 organic acid components, 11 flavonoid components and 1 coumarin component such as chicoric acid, chlorogenic acid, oroxindin, rutin, corynoline, and esculetin. In addition, 6 targets for parotitis, 48 targets for tonsillitis, and 22 targets for pharyngitis were screened. A total of 22 disease signaling pathways are involved, including 4 pathways closely related to inflammation. The IL-17 signaling pathway had the highest D(degree) value and may be most closely related to inflammatory diseases. Through network data excavating, we initially explored the main active components/ingredients of Pudilan Xiaoyan Oral Liquid, clarified the pharmacodynamic basis of Pudilan Xiaoyan Oral Liquid treatment-related diseases and its key mechanism of action in this study, hoping to provide a theoretical basis for clinical research, and at the same time, lay the foundation for deep research and promotion of Pudilan Xiaoyan Oral Liquid product.

[Improvement of compliance of Pudilan Xiaoyan Oral Liquid based on comprehensive analysis of bad taste of traditional Chinese medicine oral liquid preparation].

From "good efficacy with a bad taste" to "good efficacy with no bitterness" and then to "good efficacy with a good taste" is the only way to develop oral liquid preparations of traditional Chinese medicine, and "good medicine is beneficial to disease" is the only way for the development of oral liquid preparations of traditional Chinese medicine. Based on the analysis of the causes, the sources of bitterness, the formation principles and their solutions of traditional Chinese medicine oral liquid preparations, we explored the causes of the bad taste and the material basis of bitterness of Pudilan Xiaoyan Oral Liquid, and applied the solutions in improving the taste of Pudilan products. The overall taste of Pudilan Xiaoyan Oral Liquid was improved by modifying the original product taste, enhancing the process and changing the dosage form, which improves the compliance of the patients who take the medicine, and better serve the clinical medication.

A comparison of the pharmacokinetics of three different preparations of total flavones of Hippophae rhamnoides in beagle dogs after oral administration.

Pharmacokinetic properties of isorhamnetin, quercetin, and kaempferol in three different total flavones of Hippophae rhamnoides (TFH) preparations were compared after oral administration to beagle dogs by a UPLC-MS method. The pharmacokinetic results showed that C max of isorhamnetin and quercetin in TFH solid dispersion (TFH-SD) and TFH self-emulsifying (TFH-SE) preparations was significantly enhanced than that in TFH preparations (p < 0.05). The AUCs of isorhamnetin and quercetin in TFH-SD were 5.9- and 3.1-fold higher than that of TFH, while the AUCs of isorhamnetin and quercetin in TFH-SE were 3.4- and 2.4-fold higher than that of TFH. These findings suggested that the oral bioavailability of isorhamnetin and quercetin in beagle dogs can be significantly increased in TFH-SD and TFH-SE preparations compared to TFH preparations, which was helpful to explore the new forms for oral administration TFH and explain their in vivo processes.

Effects of stabilizing agents on the development of myricetin nanosuspension and its characterization: an in vitro and in vivo evaluation.

Although myricetin has various pharmacological applications, it shows low oral bioavailability (<10%) in rats due to its poor aqueous solubility. To overcome this issue, myricetin nanosuspensions were developed and the effects of stabilizers were investigated. Based on the particle size and zeta potential, stabilizers soya lecithin, TPGS, HP-ß-CD, and/or a combination thereof were used. The prepared nanosuspensions were characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and X-ray powder diffraction (XRD). The resulting myricetin nanosuspensions contained particles in the size range of 300-500 nm and were physically stable. Myricetin was partially transformed from crystalline to amorphous forms in the presence of different excipients after the nanosizing process. The solubility and in vitro dissolution of all myricetin nanosuspensions were greatly increased compared with those of the myricetin powder. Consequently, the relative bioavailability in rats were 2.44, 3.57, 1.61, and 2.96 for nanosuspensions stabilized with TPGS, soya lecithin, soya lecithin+TPGS, and HP-ß-CD+TPGS, respectively, relative to that of the coarse myricetin. This research demonstrated that nanosuspension is a promising strategy for delivering poor water-soluble drugs such as myricetin and that stabilizers played a critical role in the formulation design of myricetin nanosuspensions.

Effect of trichostatin A on anti HepG2 liver carcinoma cells: inhibition of HDAC activity and activation of Wnt/ß-Catenin signaling.

PURPOSE: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. MATERIALS AND METHODS: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ß-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ß-catenin, HDAC1and HDAC3 was tested by q-PCR. ß-Catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. RESULTS: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/ G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was 6.22±0.25%, which increased to 7.17±0.20% and 18.1±0.42% in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ß-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ß-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ß-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. CONCLUSIONS: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/ß-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

[Construction and expression of recombinant plasmid to initiate polarization and activation of regulatory T cells].

In order to construct a recombinant plasmid initiating the polarization and activation of the regulatory T cells (Treg), the fragments of hTGF-ß1 and C2-C4 of gp120 amplified by RT-PCR and cloned into pCR2.1 vector respectively. hTGF-ß1 and C2-C4 DNA fragments were obtained, then sub-cloned to generate the prokaryotic expression vector named pET-28a/C2-C4-Linker- hTGF-ß1. The expression of recombinant protein was induced by IPTG (0.1 mmol/L) for 6 hours. The results showed that the fragments of hTGF-ß1 and C2-C4 were amplified and cloned into pCR2.1, the prokaryotic expression vector pET-28a/C2-C4-Linker- hTGF-ß1 was constructed successfully. The recombinant protein was expressed as inclusion body after being induced by IPTG. It is concluded that this recombinant protein can initiate the polarization and activation of Treg cells, indicating the engineering E.coli strain is successfully obtained.

Construction and expression of recombinant plasmid to initiate polarization and activation of regulatory T cells / 中国实验血液学杂志

In order to construct a recombinant plasmid initiating the polarization and activation of the regulatory T cells (Treg), the fragments of hTGF-β1 and C2-C4 of gp120 amplified by RT-PCR and cloned into pCR2.1 vector respectively. hTGF-β1 and C2-C4 DNA fragments were obtained, then sub-cloned to generate the prokaryotic expression vector named pET-28a/C2-C4-Linker- hTGF-β1. The expression of recombinant protein was induced by IPTG (0.1 mmol/L) for 6 hours. The results showed that the fragments of hTGF-β1 and C2-C4 were amplified and cloned into pCR2.1, the prokaryotic expression vector pET-28a/C2-C4-Linker- hTGF-β1 was constructed successfully. The recombinant protein was expressed as inclusion body after being induced by IPTG. It is concluded that this recombinant protein can initiate the polarization and activation of Treg cells, indicating the engineering E.coli strain is successfully obtained.