Tubiechong patching promotes tibia fracture healing in rats by regulating angiogenesis through the VEGF/ERK1/2 signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The external use of traditional Chinese medicine (TCM) to treat fractures has a long history of clinical application and theoretical basis, and is also one of the characteristic treatment methods of TCM with significant efficacy and many advantages. Among the commonly used external Chinese medicines, Tubiechong is noteworthy. AIM OF THE STUDY: To elucidate whether local patching of Tubiechong can promote fracture healing and explore its mechanism of action. MATERIALS AND METHODS: A rat tibia fracture model was constructed by the modified Einhorn modeling method. X-ray films were taken to evaluate the progress of fracture healing. Serum bone alkaline phosphatase (BALP), osteocalcin (BGP) and the C-terminal content of collagen type I (CTX-I) were analyzed by ELISA. CD31 immunohistochemistry was used to evaluate angiogenesis in the tibia segment. The effects of Tubiechong decoction (TD) on HUVEC proliferation, migration and invasion were detected by MTT assay, wound healing assay and Transwell migration assay, respectively. RNA-seq was performed to identify differentially expressed genes (DEGs). Enrichment of functions and signaling pathway analysis were performed based on the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Quantitative real time polymerase chain reaction (qRT-PCR) was used to study gene expression levels. Western blotting (WB) was used to detect the expression of relevant regulatory proteins. RESULTS: The healing time of rat tibia fractures in the three TD dose groups was shortened. The serum levels of BALP, BGP and CTX- I in the TD-treated group were higher than those in the NC group. The X-ray results showed that on the 7th day after surgery, the fracture healing degree of the high-dose TD group was significantly better than that of the NC group, and the fracture healing degrees of each TD treatment group were significantly higher than those of the NC group on the 14th, 17th, and 21st days after the operation. The CD31 immunohistochemistry results showed that the number of blood vessels and the vascular area in the TD treatment group were higher than those in the NC group. In vitro, TD promoted the proliferation, wound healing and migration of HUVECs. GO analysis of transcriptome sequencing results showed that TD significantly altered the expression of genes related to cell growth, metabolism, and motility. According to KEGG annotations, VEGFA was upregulated. Eight DEGs were enriched in the VEGFA-VEGFR2 signaling pathway, of which six were upregulated. KEGG signaling pathway analysis showed that the most abundant DEGs were in mitogen-activated protein kinase (MAPK) signaling pathway. qRT-PCR showed that VEGFA gene expression in HUVECs was 7.8 times that of the control group after 1 mg/mL TD treatment for 24 h, and WB experiments showed that its protein expression was 3 times that of the control group. WB results showed that the phosphorylated ERK gene was highly expressed, while the expression levels of phosphorylated P38 and phosphorylated JNK protein remained unchanged. CONCLUSION: Tubechong patching therapy promotes tibia fracture healing in rats by regulating angiogenesis through the VEGF/ERK1/2 signaling pathway.

Tubiechong：A review on ethnomedicinal uses, bioactive chemical constituents and pharmacological activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Tubiechong comprises mainly Eupolyphaga and Steleophaga is widely distributed in China. It has been used in the traditional medicine systems in Asian countries specially in China，Japan and Singapore for thousand years. AIM OF THE REVIEW: The aim of this work is to review the scientific work about Tubiechong regarding their ethnomedicinal uses, bioactive chemical constituents and pharmacological activities. MATERIALS AND METHODS: Relevant literature of Tubiechong was collected for its traditional uses, pharmacological activities, and bioactive compounds released from inception until May 2022. The online databases such as Web of Science, PubMed, Google Scholar, Science Direct, Scopus, SciFinder Scholar, Springer Link, China National Knowledge Infrastructure (CNKI), Wanfang Data, and VIP database were used as electronic search engines for articles with the various specific keywords. Additionally, references from ancient texts and local information such as PhD and MSc theses, books, and Chinese journals were also included. RESULTS: The clinical researches have revealed that Tubiechong alone has been successfully used to treat bone disease, ache, sprain, herpes zoster, paronychia and so on. Tubichong's main clinical application is to form formulations with other herbs. The most widely used 34 kinds of Chinese patent medicine containing Tubiechong were included in Chinese Pharmacopoeia (2020 Edition) for the treatment of traumatic injury, low back pain, cardiovascular disease, tumors or mass and nodule, cervical spondylopathy, osteoarthritis and psoriasis. Its other derived formulas have been used in the clinical treatment of various diseases, such as blood stasis, hepatic cirrhosis, cyclomastopathy, chronic active hepatitis, nephropathy, gynaecopathia, cancer diseases. To date, the bioactive substances reported are limited to protein and peptides, fatty acids, polysaccharides and alkaloids from Eupolyphaga sinensis Walker. So far, the pharmacological activities of Tubiechong and its various extracts have been evaluated, including anticoagulant and antithrombotic, anticancer, bone repair, immunomodulation, analgesia, antioxidant, antihyperlipidemic, antimicrobial and protective and repair functions for damage to the liver, heart, brain and skin. As an edible insect, its safety has also been confirmed by acute toxicity tests and 30-day feeding trials. CONCLUSION: Tubiechong is an important insect medicine with the effect of promoting blood circulation and removing blood stasis, which has been used in traditional Chinese medicine for thousands of years for the treatment of trauma and abdominal lumps, and has now been clinically extended to the treatment of a variety of diseases. Its multiple pharmacological activities indicate that it has great potential for development and application. However, its chemical constituents with pharmacological activity require further excavation and detailed study. In addition, the in-depth molecular pharmacological mechanisms deserve further explanation.

Efficacy of different nutrients in age-related macular degeneration: A systematic review and network meta-analysis.

AIM: Age-related macular degeneration (AMD) has become a predominant global health concern. The visual function of individuals with AMD seems to improve with dietary antioxidants. We assessed the efficacy of different antioxidants (carotenoids, zinc, vitamin E, and multivitamin) on visual function and the incidence of developing late AMD. METHODS: We searched PubMed, EMBASE, and Cochrane Central Register of Controlled Trials for related published studies. We considered randomized controlled trials (RCTs) comparing different nutrients. The main outcomes measurements included changes in visual acuity (VA), and the rate of developing late AMD. The network meta-analysis was registered on PROSPERO (CRD42020171288). RESULTS: We identified 13 studies, including 85321 individuals randomly assigned to different nutrients or placebo groups. In the network meta-analysis, we found that there was more risk of progression to late AMD in the multivitamin group than carotenoids and vitamin E groups (RR 0.45, 95% CI 0.32 to 0.65; RR 0.56, 95% CI 0.40 to 0.79; RR 0.42, 95% CI 0.26 to 0.67). The nutrients of zinc and carotenoids (Lutein/Zeaxanthin) ranked first and second and showed better improvement in VA. The efficacy of carotenoids (ß-carotene) ranked first for delaying the progress of AMD among all of the four treatments. CONCLUSION: Taking multivitamin supplementation may not prevent the development of late AMD. The nutrient of zinc and carotenoids (lutein/zeaxanthin) supplementation were associated with better improvement in VA. Carotenoids (ß-carotene) were the most likely to prevent the progression of late AMD.

Xuebijing Injection Ameliorates H2S-Induced Acute Respiratory Distress Syndrome by Promoting Claudin-5 Expression.

OBJECTIVE: To investigate the protective effects and underlying mechanisms of Xuebijing Injection (XBJ) on the lung endothelial barrier in hydrogen sulfide (H2S)-induced acute respiratory distress syndrome (ARDS). METHODS: Sprague-Dawley rats were exposed to H2S (300 ppm) to establish ARDS model, while human pulmonary microvascular endothelial cells (HPMECs) were incubated with NaHS (a H2S donor, 500 µmol/L) to establish cell model. H2S and XBJ were concurrently administered to the rat and cell models. Lung hematoxylin and eosin staining, immunohistochemistry, transmission electron microscopy and wet/dry ratio measurement were used to confirm ARDS induced by H2S in vivo. The expression levels of claudin-5, phosphorylated protein kinase B (p-AKT)/t-AKT and p-forkhead box transcription factor O1 (FoxO1)/t-FoxO1 in vivo and in vitro were also assessed. Paracellular permeability and transepithelial electrical resistance (TEER) were measured to evaluate endothelial barrier function in the cell model. RESULTS: The morphological investigation showed that XBJ attenuated H2S-induced ARDS in rats. XBJ significantly ameliorated both the reduction in TEER and the increased paracellular permeability observed in NaHS-treated HPMECs (P<0.05). The protective effects of XBJ were blocked by LY294002, a phosphatidylinositol 3-kinase (PI3K)/AKT/FoxO1 pathway antagonist (P<0.05). Furthermore, XBJ promoted the expression of claudin-5 and increased the levels of p-AKT and p-FoxO1 in vivo and in vitro (P<0.05). CONCLUSIONS: XBJ ameliorated H2S-induced ARDS by promoting claudin-5 expression via the PI3K/AKT/FoxO1 signaling pathway.

Lycibarbarines A-C, Three Tetrahydroquinoline Alkaloids Possessing a Spiro-Heterocycle Moiety from the Fruits of Lycium barbarum.

Three tetrahydroquinoline alkaloids, lycibarbarines A-C (1-3), possessing a unique tetracyclic tetrahydroquinoline-oxazine-ketohexoside fused motif, were isolated from the fruits of Lycium barbarum. Their structures were determined by spectroscopic analysis and quantum-chemical calculations. Compounds 1 and 3 exhibited neuroprotective activity when evaluated for corticosterone-induced injury by reducing the apoptosis of PC12 cells through the inhibition of caspase-3 and caspase-9.

[Effect of Dilong on expression of fibrogenic factors TGF-ß1 and α-SMA in lung tissue of mice with pulmonary fibrosis].

The aim of this paper was to investigate the effect of Dilong( geosaurus) on the expressions of fibrotic factors TGF-ß1 and α-SMA in bleomycin-induced pulmonary fibrosis mice. The binding ability of Dilong to fibrotic factor TGF-ß1 was initially detected by Biacore technology and verified by in vivo pharmacodynamics. A total of 60 SPF C57 mice were randomly divided into 6 groups. Except the blank group( injecting 0. 08 m L·kg-1 sodium chloride in the trachea),the other five groups were given bleomycin( 4 mg·kg-1) to replicate the pulmonary fibrosis model. After 14 days of drug treatment,the expressions of TGF-ß1 and α-SMA were detected by Masson staining,immunohistochemistry and RT-PCR. The results of Biacore experiment showed that the extract of Dilong was well bound to TGF-ß1 protein in vitro,and the binding value reached 619. 3. Compared with the model group,Masson's results showed that cellulose deposition in high-dose,medium-dose and low-dose Dilong groups decreased to varying degrees. RT-PCR results showed that different doses of Dilong could reduce protein and mRNA expressions of TGF-ß1 and α-SMA to a certain extent in a dose-dependent manner. In conclusion,Dilong could delay the process of pulmonary fibrosis by binding to target protein TGF-ß1 and inhibiting the expression of α-SMA.

Prenatal High Estradiol Exposure Induces Sex-Specific and Dietarily Reversible Insulin Resistance Through Decreased Hypothalamic INSR.

An adverse intrauterine environment may induce adult disease in offspring, but the mechanisms are not well understood. It is reported that fresh embryo transfer (ET) in assisted reproductive technology leads to high maternal estradiol (E2), and prenatal high E2 exposure increases the risk of organ disorders in later life. We found that male newborns and children of fresh ET showed elevated fasting insulin and homeostasis model of assessment for insulin resistance index (HOMA-IR) scores. Male mice with high prenatal estradiol exposure (HE) grew heavier than control mice and developed insulin resistance; they also showed increased food intake, with increased orexigenic hypothalamic neuropeptide Y (NPY) expression. The hypothalamic insulin receptor (INSR) was decreased in male HE mice, associated with elevated promoter methylation. Chronic food restriction (FR) in HE mice reversed insulin resistance and rescued hypothalamic INSR expression by correcting the elevated Insr promoter methylation. Our findings suggest that prenatal exposure to high E2 may induce sex-specific metabolic disorders in later life through epigenetic programming of hypothalamic Insr promoter, and dietary intervention may reverse insulin resistance by remodeling its methylation pattern.

[Mechanism of anti-aging therapy on Parkinson's disease].

Many studies have shown that anti-aging treatment has value to prevention and treatment of some diseases. For the treatment of Parkinson' s disease, clinical and experimental researches have proved the potential value of anti-aging treatment, yet the mechanism remains unclear. For this reason, this work used the anti-aging prescriptions of Buyang Huanwu decoction in traditional Chinese medicines example to discover the anti-aging treatment mechanism on Parkinson's disease. The results showed that the mechanism of mitochondrial damage, apoptosis, free radicals and oxidative stress could contribute to the treatment of Parkinson' s disease. Buyang Huanwu decoction is more than as the carrier in this article, the discovered anti-aging treatment mechanism Parkinson's disease is not confined to Buyang Huanwu decoction, could also be used to understand the anti-aging treatment mechanism using other prescription. The main contribution of this paper is to clarify the mechanism of anti-aging treatment of Parkinson's disease, and provide a new strategy for the treatment and prevention of Parkinson's disease.

[Effect of Qige Powder on Angiogenesis Induced by Esophageal Cancer Cell Line Eca-9706].

OBJECTIVE: To study the effect Qige powder on esophageal cancer angiogenesis. METHODS: Inhibitive effect of Qige powder ethanol extract on proliferation of Esophageal cancer cell line Eca-9706 was detected by MTT assay. The Eca-9706 cells conditioned medium in Qige powder IC50 concentration were collected. Angiogenesis, as well as proliferation, migration and tube formation of human umbilical vein endothelial cells were observed by Chick embryo chorioallantoic membrane model(CAM), MTT assay, the migration and tube formation assay. VEGF and IL-6 contents in culture medium supernatant of Eca-9706 cells and human umbilical vein endothelial cells were detected by ELISA. RESULTS: Qige powder ethanol extract could inhibit the proliferation of Eca-9706 cells, with a certain dose-effect relationship, IC50 value was 96 µg/mL. Eca-9706 cells conditioned medium could significantly increase the CAM generating total vessel area, human umbilical vein endothelial cells proliferation,migration and tube formation, while the Eca-9706 cell conditioned medium of Qige powder ethanol extract could reduce CAM angiogenesis, human umbilical vein endothelial cells proliferation and tube formation, but increase endothelial cells migration. Qige powder ethanol extract could reduce endothelial cells secreting VEGF and IL-6 while increase Eca-9706 cells secreting. CONCLUSION: Qige powder ethanol extract can inhibit angiogenesis, endothelial cell proliferation, and tube formation induced by Eca-9706 cells, while reduce VEGF and IL-6 secretion of endothelial cells.

[Study on mechanism of Salvia miltiorrhiza treating cardiovascular disease through auxiliary mechanism elucidation system for Chinese medicine].

Salvia miltiorrhiza is a traditional Chinese medicine (TCM) and is widely used as a clinically medication for its efficiency in treating cardiovascular disease. Due to TCM is a comprehensive system, the mechanism of S. miltiorrhiza treating cardiovascular disease through integrated multiple pathways are still unclear in some aspects. With the rapid progress of bioinformatics and systems biology, network pharmacology is considered as a promising approach toward reveal the underlying complex relationship between an herb and the disease. In order to discover the mechanism of S. miltiorrhiza treating cardiovascular disease systematically, we use the auxiliary mechanism elucidation system for Chinese medicine, built up a molecule interaction network on the active component targets of S. miltiorrhiza and the therapeutic targets of cardiovascular disease to offer an opportunity for deep understanding the mechanism of S. miltiorrhiza treating cardiovascular disease from the perspective of network pharmacology. The results showed that S. miltiorrhiza treating cardiovascular disease through ten pathways as follows: improve lipid metabolism, anti-inflammation, regulate blood pressure, negatively regulates blood coagulation factor and antithrombotic, regulate cell proliferation, anti-stress injury, promoting angiogenesis, inhibited apoptosis, adjust vascular systolic and diastolic, promoting wound repair. The results of this paper provide theoretical guidance for the development of new drugs to treat cardiovascular disease and the discovery of new drugs through component compatibility.

[Study on mechanism for anti-hypertension efficacy of Eucommiae Cortex through assistant analysis systems for acting mechanisms of traditional Chinese medicine].

Elucidate the mechanism of Eucommiae Cortex in treatment of hypertension, to provide the basis for further research and development of Eucommiae Cortex. Our study using the entity grammar systems inference rules to analyse the interactions of chemical constituents of Eucommiae Cortex and disease target proteins at the molecular level, and got a biological network of Eucommiae Cortex anti-hypertension which inciude 602 nodes and 2 354 edges. We got 3 treatment of hypertension pathways of Eucommiae Cortex by analyzing biological network, that is, by inhibition of vascular remodeling to improve the deterioration of hypertension, reduce activity of polymorphism genetic genes related to essential hypertension, inhibition of carbonic anhydrase 1 to maintain the osmotic pressure, Eucommiae Cortex play the role of anti-hypertension.

[Study on mechanism for anti-depression efficacy of Sini San through auxiliary mechanism elucidation system for Chinese medicine].

As regulating the function of the liver and spleen of the famous traditional formula, Sini San is widely used in the treatment of various diseases caused by liver depression and Qi stagnation, and its efficacy is significant clinically. Recently it is discovered that Sini San is effective in the treatment of nervous system diseases such as depression. Furthermore, there is a lot of literature about the effect of Sini San on the molecular mechanism of antidepressant. However, the anti-depression mechanism of Sini San is not very clear, in our present study, based on the auxiliary mechanism elucidation system for Chinese medicine and network pharmacology system to construct the chemical ingredients of the target interactions and disease-related protein of the interaction network. Results show that there are 263 chemical ingredients and 19 corresponding targets of depression in Sini San network. Sini San can anti-depressant effect through G-protein coupled receptor protein signaling pathway, cAMP system, neurological system process and neurotransmitter secretion, inflammatory response, neuroendocrine, metal ion transport and so on. These studies provided valuable clues for the mechanism and treatment of anti-depressant.