Background A new pathogenic role for mitochondrial dysfunction has been associated with aging and correlated with the development of idiopathic pulmonary fibrosis (IPF). The latest study found that the lysine succinylation (Ksucc) is involved in many energy metabolism pathways and affects the metabolic process in mitochondria, making this modification highly valuable for studying IPF related to mitochondrial dysfunction. We speculate Ksucc participate in IPF progression through mitochondrial energy metabolism pathway. Methods We used liquid chromatography with tandem mass spectrometry (LC-MS/MS) to perform the first global profiling of Ksucc in lung tissues with IPF patients. The changes of candidate key proteins and Ksucc sites related to energy metabolism in IPF lung tissues were analyzed by using the clusters of orthologous groups of proteins (COG), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene ontology (GO). We then compared these proteins with those reported in the literature in normal lung tissues by parallel reaction monitoring (PRM). Results We identified 1964 Ksucc sites in 628 proteins. 675 Ksucc sites in 124 proteins closely related to mitochondrial metabolism. We compared these proteins with those reported in the literature in normal lung tissues to identify differences in 119 proteins and Ksucc sites in mitochondria. 43 Ksucc sites in 27 proteins were associated with energy metabolism. There were differences in the expression of 4 Ksucc sites in 4 proteins between normal and IPF lung tissues. Conclusion Our work expands the Ksucc database in IPF lung and suggested that mitochondrial energy metabolism is involved in the progression of IPF. Ksucc sites of proteins associated with mitochondrial energy metabolism can also serve as candidate molecules for future mechanism exploration and drug target selection in IPF.

BACKGROUND:Intracerebral hemorrhage (ICH) is a common cerebrovascular disease, and the complement cascade exacerbates brain injury after ICH. As the most abundant component of the complement system, complement component 3 (C3) plays essential roles in all three complement pathways. However, the effects of C3 on neurological impairment and brain injury in ICH patients and the related mechanism have not been fully elucidated. Normobaric hyperoxia (NBO) is regarded as a treatment for ICH patients, and recent clinical studies also have confirmed the neuroprotective role of NBO against acute ICH-mediated brain damage, but the underlying mechanism still remains elusive.AIMS:In the present study, we investigated the effects of complement C3 on NBO-treated ICH patients and model mice, and the underlying mechanism of NBO therapy in ICH-mediated brain injury.RESULTS:Hemorrhagic injury resulted in the high plasma C3 levels in ICH patients, and the plasma C3 levels were closely related to hemorrhagic severity and clinical outcomes after ICH. BO treatment alleviated neurologic impairments and rescued the hemorrhagic-induced increase in plasma C3 levels in ICH patients and model mice. Moreover, the results indicated that NBO exerted its protective effects of on brain injury after ICH by downregulating the expression of C3 in microglia and alleviating microglia-mediated synaptic pruning.CONCLUSIONS:Our results revealed that NBO exerts its neuroprotective effects by reducing C3-mediated synaptic pruning, which suggested that NBO therapy could be used for the clinical treatment of ICH.

Background: The Danshen-Honghua herb pair (DHHP) is a common modern Chinese medicine pair for activating blood circulation and resolving blood stasis. It has been used for centuries to treat cardiovascular and cerebrovascular diseases and is often found in some herbal compounds for treating cardiovascular diseases. The aim of this study was to explore the effects of DHHP on the intestinal flora of rats with acute myocardial ischemia and its microbial transformation in vitro. Methods: In this study, we investigated the protective effect of DHHP on isoproterenol-induced acute myocardial ischemia in rats based on metagenomic sequencing technology, and further characterized the in vitro metabolic transformation products of DHHP, so as to investigate its anti-myocardial ischemic efficacy material basis. Results: Pharmacodynamic results demonstrated that DHHP significantly ameliorated pathological changes and improved abnormal cardiac enzyme levels in acute myocardial ischemia rats. In addition, metagenomic analysis showed the efficacy of DHHP in ameliorating the isoproterenol-induced modifications of the intestinal flora in rats. Specifically, DHHP promoted the growth of the intestinal potential probiotics such as Lactobacillus while suppressing the pathogenic bacteria, including Escherichia and Streptococcus. The in vitro metabolism results showed that the DHHP's active components underwent primarily phase I metabolism through hydroxylation, decarboxylation, and dehydration inversions in the isolated intestinal flora of acute myocardial ischemia rats and in phase II through sulfation esterification and methylation reactions. Conclusion: The results suggest that there may be a bidirectional regulatory effect between DHHP and intestinal flora, which is important to explain the pharmacological mechanism of DHHP.

目的 采用系统预试法对缘毛太行花的化学成分进行预实验,初步探索缘毛太行花的化学成分类别.方法 采用乙醇、三氯甲烷、甲醇等不同溶剂对缘毛太行花进行提取,提取液A中分别加入氢氧化钠溶液,乙酸钾溶液、三氯化铁溶液、浓硫酸没食子酸溶液、碘化铋钾试剂、碘-碘化钾试剂、盐酸-镁粉:提取液B分别加入浓硫酸,浓硫酸-乙酸酐溶液观察有无阳性反应的发生;在试管反应的基础上采用薄层色谱鉴别及高效液相色谱法对其中的三萜类成分进一步进行定性鉴别.结果 缘毛太行花的提取液加氢氧化钠溶液变为红色、加乙酸钾溶液变为橙黄色、加三氯化铁溶液为墨绿色、加入浓硫酸-没食子酸溶液无明显变化、加入浓硫酸有绿色荧光、加浓硫酸-乙酸酐产生黄-蓝的颜色变化、加入碘化铋钾生成橘红色沉淀、加入碘-碘化钾溶液生成红棕色无定形沉淀;薄层色谱鉴别样品和对照品在同一处显示相同颜色的斑点;高效液相色谱法显示样品和对照品在相同保留时间出峰.结论 初步确定缘毛太行花中包含酚羟基类官能团的特征以及蒽醌类、三萜类及生物碱类等多种化学成分,为缘毛太行花的研究奠定基础.

MALAT1 is one of the most hopeful members implicated in angiogenesis in a variety of non-malignant diseases. In multiple myeloma (MM), MALAT1 is recognized as the most highly expressed long non-coding RNA. However, the functional roles of MALAT1 in angiogenesis and the responsible mechanisms have not yet been explored. Herein, we discovered a novel regulatory network dependent on MALAT1 in relation to MM tumorigenesis and angiogenesis. We observed that MALAT1 was upregulated in MM and significantly associated with poor overall survival. MALAT1 knockdown suppressed MM cell proliferation and promoted apoptosis, while restricting endothelial cells angiogenesis. Moreover, MALAT1 directly targeted microRNA-15a/16, and microRNA-15a/16 suppression partly reverted the effects of MALAT1 deletion on MM cells in vitro as well as tumor growth and angiogenesis in vivo. In addition, further study indicated that MALAT1 functioned as a competing endogenous RNA for microRNA-15a/16 to regulate vascular endothelial growth factor A (VEGFA) expression. Our results suggest that MALAT1 plays an important role in the regulatory axis of microRNA-15a/16-VEGFA to promote tumorigenicity and angiogenesis in MM. Consequently, MALAT1 could serve as a novel promising biomarker and a potential antiangiogenic target against MM.

Abstract Syringa oblata is a flowering tree endemic to China, which has high medicinal and economic value. Comparative genomic analysis is necessary to elucidate the variation and phylogenetic relationships among Syringa species. An Illumina NovaSeq 6000 sequencing platform was used to sequence the whole genome of Syringa oblata plastomes in this study, and sequence assembly, annotation and characterization were performed by bioinformatics analysis, and a phylogenetic tree using Maximum Likelihood (ML) was constructed. A total of 155,668 bp was contained in the genome, which comprised two inverted repeats (IR) of 25,733 bp each, separated by a small single copy (SSC) region of 17,926 bp and a large single copy (LSC) region of 86 bp. There is a total GC content of 37.9% in the chloroplast genome, while the GC content in each region of SSC, LSC, and IR is 36.0%, 32.1%, and 43.2%. The genome contains 132 genes, including 88 protein-coding genes (CDS), 36 tRNA genes and 8 rRNA. There are seven protein-coding genes in the IR region, seven tRNA genes, and four rRNA genes. There was the most abundance of leucine and the least abundance of tryptophan among all amino acids. We detected 36 simple sequence repeats (SSRs) in the repeat structure analysis which included 32 mononucleotides (p1), 1 dinucleotide (p2), 1 trinucleotide (p3), 1 tetranucleotides (p4), and 1 complex nucleotide were explored, in the S. oblata chloroplast genome. In addition, we identified 50 long repeats, comprising 21 forward repeats, 28 palindromic repeats, and 1 complementary repeat. A phylogenetic analysis suggested that S. oblata (MW464119) is closely related to Syringa vulgaris (MG255768). It is anticipated that our results will be useful to further species identification, population genetic analysis, and biological research on Syringa oblata in the future.

OBJECTIVE:This study aimed to investigate the active components and mechanism of action of rosemary volatile oil for treating Alzheimer's disease (AD) using network pharmacology.METHODS:We obtained the constituents of the rosemary volatile oil by searching Chinese herbal systemic pharmacological databases and analytical platforms and constructed the constituent-target networks by predicting and screening the action targets of the rosemary volatile oil constituents using SwissTargetPrediction, metaTarFisher, and Pubchem. We obtained the AD-related targets using the Genecards, OMIM, and DisGeNET databases and constructed the protein-protein interaction networks (PPI) using the STRING database in Venny 2.1.0 graph to identify the cross-targets by screening the core-acting targets. Cytoscape 3.8.2 software was used to construct a componenttarget- pathway network to screen the potential active components of the rosemary volatile oil for the treatment of AD and predict the mechanism of action of the rosemary volatile oil for the treatment of AD in combination with existing pharmacological studies. We performed a gene ontology (GO) biological process and a Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the targets of the rosemary volatile oil for the treatment of AD using R language and molecular docking using Discovery Studio 4.0 software to validate their biological activities.RESULTS:A network constructed using gas chromatography-mass spectrometry (GC-MS) analysis identified 26 potentially active ingredients in the rosemary volatile oil. We retrieved a total of 10762 AD targets from Genecards and other databases. Our GO enrichment analysis yielded 39 entries (P < 0.05), including 14 entries for biological processes, five entries for cellular composition, and 20 entries for molecular function. A total of 14 entries (P < 0.05) were then enriched in the KEGG pathway that primarily involved the IL-17 signaling pathway and the AGE-RAGE pathway.CONCLUSION:The active components of rosemary volatile oil had good inhibition of the inflammatory response. This study provides a reference and guidance for the in-depth study on rosemary volatile oil for the treatment of AD.

In recent years, more and more evidence has shown that the disorder of gut microbiota (GM) is closely correlated with myocardial ischemia (MI). Even though the Danshen and Honghua herb pair (DHHP) is widely used in treating cardiovascular disease in China and exhibits obvious clinical efficacy on MI, the anti-MI mechanism of DHHP remains and needs to be explored in depth. Thus, in this study, we investigated whether the amelioration effect and molecular mechanism of DHHP on MI were related to regulating GM through pharmacodynamics evaluation and metagenomic sequencing. Histopathological testing results showed that DHHP treatment could alleviate the pathological changes of myocardial tissue in the acute MI (AMI) rats induced by isoproterenol (ISO), especially structural disorder, irregular distribution, and enlargement of the myocardial space. These pathological changes were all alleviated to some extent by DHHP treatment. Biochemical analysis results suggested that compared with the control group, the serum levels of AST, CTn-I, CK-MB, and TNF-alpha in model group rats were notably decreased, and the CAT and SOD levels in serum were markedly increased. These abnormal trends were significantly reversed by DHHP treatment. Furthermore, metagenomic sequencing analysis results indicated that DHHP could improve disorders in the composition and function of GM in AMI rats, mainly reflected in increasing diversity and richness, and obviously enhancing the abundance of Bacteroides fluxus, B. uniformis, B. stercoris, Roseburia hominis, Schaedlerella arabinosiphila, and R. intestinalis, and reducing the abundance of Enterococcus avium and E. canintestini, which were associated with purine metabolism, tyrosine metabolism, cyanoamino acid metabolism, and glutathione metabolism. In conclusion, DHHP may attenuate ISO-induced MI by regulating the structure, composition, and function of GM, thus contributing to further our understanding of the anti-MI mechanisms of DHHP and providing new therapeutic ideas and diagnostic targets for the clinical studies of MI. Danshen and Honghua herb pair could improve the disorder of the composition and function of intestinal flora in rats with myocardial ischemic injury.

Recent studies reported that advanced glycation end products (AGEs) and endothelial-to-mesenchymal transition (EndMT) were involved in the calcific aortic valve disease (CAVD). However, the roles of AGEs in EndMT in the development of CAVD have not been elucidated. In this study, six-week-old male Apoe(-/-) mice were divided into four groups based on the following feeding periods: 0, 2, 4, and 6 months. The latter three groups were further divided into three subgroups corresponding to the following diet treatments: normal diet, high-fat diet + normal saline injection, and high-fat diet + aminoguanidine injection. Weight gain was monitored weekly. Finally, heart echocardiographic assessments were performed, and serum lipid levels, the protein expression and the histological changes in the aortic valves were determined. Results showed that the AGE inhibitor aminoguanidine alleviated the transaortic valve velocity and decreased the total cholesterol and low-density lipoprotein cholesterol levels. Calcification and carboxymethyl-lysine deposition were firstly detected around the aortic valve surfaces, whereas aminoguanidine injection attenuated their accumulation. In the early stage, HFD-activated AGEs-RAGE signaling resulted in the alpha-smooth muscle actin (alpha-SMA) upregulation and the vascular endothelium cadherin (VE-cadherin) downregulation on the valve endothelial layer. The activation resulted in early the transforming growth factor-beta 1 (TGF-beta 1) and the alkaline phosphatase (ALP) upregulation, and simultaneously reduced the bone morphogenetic protein receptor type II (BMPR2) expression. However, aminoguanidine restricted these proteins changes by inhibiting the interaction of AGEs and RAGE. In addition, immunofluorescence images showed obvious double-positive staining of ALP and a-SMA on the valve surfaces, revealing the contribution of EndMT to the early calcification. Therefore, this study demonstrates that activation of the AGEs-RAGE axis induced EndMT, promoting the progression of the aortic valve calcification in the initial stage via the counteraction of BMPR2 and TGF-beta 1 signaling.

Atrial fibrillation (AF) affects >30 million individuals worldwide. However, no genetic mutation from human patients with AF has been linked to inflammation. Here, we show that AF-associated human variant p.Ile138Thr in natriuretic peptide A (NPPA) encoding the atrial natriuretic peptide (ANP) causes inflammation, fibroblast activation, atrial fibrosis, and AF in knock-in (KI) rats. Variant p.Ile138Thr inhibits the interaction between ANP and its receptor natriuretic peptide receptor A and reduces intracellular cGMP levels. RNA sequencing and follow-up analyses showed that mutant ANP (mANP) activates multiple innate immunity pathways, including TNF-α, NF-κB, and IL-1β signaling. mANP induces differentiation of cardiac fibroblasts (CFs) to myofibroblasts and promotes CF proliferation and fibrosis. These results suggest that NPPA variant p.Ile138Thr causes AF by activating TNF-α, NF-κB, and IL-1β signaling, inflammation, and fibrosis. Multiple computational programs suggest that p.Ile138Thr is damaging or deleterious. Based on the 2015 American College of Medical Genetics and Genomics Standards and Guidelines, p.Ile138Thr can be classified as a likely pathogenic variant. Variant p.Ile138Thr was found only in Asian people in the Genome Aggregation Database and Exome Aggregation Consortium database at an averaged frequency of 0.026%. An estimated 1.15 million Asian people carry the variant and might be at risk of AF. The KI rats may provide an inflammation-based, genetic animal model for AF valuable for testing anti-inflammation or other therapies for AF.-Cheng, C., Liu, H., Tan, C., Tong, D., Zhao, Y., Liu, X., Si, W., Wang, L., Liang, L., Li, J., Wang, C., Chen, Q., Du, Y., Wang, Q. K., Ren, X. Mutation in NPPA causes atrial fibrillation by activating inflammation and cardiac fibrosis in a knock-in rat model.

Microglia play an essential role during cerebral an ischemia/reperfusion (I/R)-related inflammatory process. Because the M2 phenotype of microglia exhibits anti-inflammation activity, it has become a promising target for anti-inflammatory therapy. Vagus nerve stimulation (VNS) reportedly has neuroprotective effects against cerebral I/R injuries via its anti-inflammatory action. The aim of this study was to investigate the ability of non-invasive VNS (nVNS) to alleviate cerebral I/R in mice by promoting microglial M2 polarization. Neurological scoring and cerebral infarct volume assessments were performed 72 h after a middle cerebral artery occlusion (MCAO)-induced stroke. M2 phenotype microglia were identified by immunohistochemistry staining using Arg-1 and Iba-1 antibodies. The protein expressions of Arg-1, IL-17A, IL-10, Bax, and Bcl-2 were detected by Western blot. Apoptotic cells were detected using TUNEL staining. According to our results, nVNS decreased infarct volume, improved neurological outcomes, reduced apoptotic neurons (TUNEL+NeuN+ cells), and promoted microglial M2 polarization as indicated by elevated Arg-1 protein expression and increased Arg-1+ cells after MCAO. Moreover, nVNS attenuated the increased levels of IL-17A protein expression after MCAO. To test the possible involvement of IL-17A in nVNS-induced neuroprotection and microglial M2 polarization, 1-μg recombinant IL-17A (rIL-17A) was intranasally administered once daily for three consecutive days after reperfusion. We found that the intranasal administration of rIL-17A nullified the nVNS-induced promotion of microglial M2 polarization. Furthermore, rIL-17A administration abolished the neuroprotective effect of nVNS. In conclusion, our study identifies microglial M2 polarization as an important mechanism underlying the nVNS-mediated neuroprotection against cerebral I/R. This effect of nVNS could be attributed to the inhibition of IL-17A expression.

Effect of hysteroscopic adhesiolysis combined with growth hormone on endometrial blood flow and volume as well as Smad2/3 expression Xing-Chan Li, Cui-Xia Liang, Jing Li, Pei-Feng Li, Yu Zhao Department of Gynecology, Tangxia Hospital of Dongguan Guangdong Province, Dongguan City, Guangdong Province, 523710, China Cross Infection Control Office, Tangxia Hospital of Dongguan Guangdong Province, Dongguan City, Guangdong Province, 523710, China Journal of Hainan Medical University

OBJECTIVE To evaluate anti-tumor effects of 2-(1-hydroxy-4-oxo-cyclohexa-2,5-dienyl)-pyran-4-one(RY10-4)against human breast cancer xenograft(MCF-7)in nude mice and investigate possible mechanisms.METHODS Nude mice xenograft models were established with MCF-7 human breast cancer cells and mice were randomized into 4 groups:model control group,low dose of RY10-4 group(7.5 mg·kg-1),high dose of RY10-4 group(15 mg·kg-1)and paclitaxel group(15 mg·kg-1).All mice in different groups were treated by intraperitoneal injections every other day,and tumor volumes and body weights of mice were detected.After treated for 16 days,all animals were killed and tumors were weighed;apoptosis index was detected by TUNEL and protein expressions of Bcl-2,Bax and MAPK pathway members were detected by western blot in tumors in each group.RESULTS Compared with model group,tumor volumes and weights of all drugs treated groups were significantly decreased,and tumor inhibition rates in low dose of RY10-4 group,high dose of RY10-4 group and paclitaxel group were 29.8%,47.2%and53.8%,respectively.TUNEL showed that apoptotic cells in all drugs treated groups were obviously increased(P0.01).Western blot showed that RY10-4 decreased anti-apoptotic protein expression of Bcl-2 increased pro-apoptotic protein expression of Bax in a time dependent manner,decreasing Bcl-2/Bax ratio(P0.05);and it also showed RY10-4 increased p-p38 and p-ERK protein levels compared with control group.CONCLUSION RY10-4 is able to inhibit growth of breast cancer xenograft in nude mice significantly and shows activity of inducing apoptosis,and mechanisms might include decreasing Bcl-2 and increasing Bax,changing Bcl-2/Bax ratio and activating p38 and ERK signal pathways.