Rapid and accurate quality evaluation of Angelicae Sinensis Radix based on near-infrared spectroscopy and Bayesian optimized LSTM network.

The authentic traditional Chinese medicines (TCMs) including Angelicae Sinensis Radix (ASR) are the representative of high-quality herbals in China. However, ASR from authentic region being adulterated or counterfeited is frequently occurring, and there is still a lack of rapid quality evaluation methods for identifying the authentic ASR. In this study, the color features of ASR were firstly characterized. The results showed that the authentic ASR cannot be fully identified by color characteristics. Then near-infrared (NIR) spectroscopy combined with Bayesian optimized long short-term memory (BO-LSTM) was used to evaluate the quality of ASR, and the performance of BO-LSTM with common classification and regression algorithms was compared. The results revealed that following the pretreatment of NIR spectra, the optimal NIR spectra combined with BO-LSTM not only successfully distinguished authentic, non-authentic, and adulterated ASR with 100 % accuracy, but also accurately predicted the adulteration concentration of authentic ASR (R2 > 0.99). Moreover, BO-LSTM demonstrated excellent performance in classification and regression compared with common algorithms (ANN, SVM, PLSR, etc.). Overall, the proposed strategy could quickly and accurately evaluate the quality of ASR, which provided a reference for other TCMs.

Effects of Diosmin on Vascular Leakage and Inflammation in a Mouse Model of Venous Obstruction.

At present, iliac vein compression syndrome (IVCS) plagues countless people, posing a significant economic and social burden. The progress of current IVCS-related research is slow owing to the limitations of animal models. In this study, we generated a mouse model of iliac vein stenosis (IVS) to monitor the effects of IVCS on venous function, such as increased vascular leakage, the expression of adhesion molecules, and elevated inflammation factors. Diosmin, a widely used clinical bioactive ingredient, was administered to confirm its therapeutic effects on the IVS mouse model. The results revealed that diosmin manifested therapeutic improvement in the IVS mouse model. In addition, we verified that the IVS mouse model is a stable and reproducible animal model for pathophysiological studies. High-purity diosmin can be beneficial to venous dysfunction and hence provides a more effective treatment option for venous diseases.

Immune metabolism: a bridge of dendritic cells function.

An increasing number of researches have shown that cell metabolism regulates cell function. Dendritic cells (DCs), a professional antigen presenting cells, connect innate and adaptive immune responses. The preference of DCs for sugar or lipid affects its phenotypes and functions. In many diseases such as atherosclerosis (AS), diabetes mellitus and tumor, altered glucose or lipid level in microenvironment makes DCs exert ineffective or opposite immune roles, which accelerates the development of these diseases. In this article, we review the metabolism pathways of glucose and cholesterol in DCs, and the effects of metabolic changes on the phenotype and function of DCs. In addition, we discuss the effects of changes in glucose and lipid levels on DCs in the context of different diseases for better understanding the relationship between DCs and diseases. The immune metabolism of DCs may be a potential intervention link to treat metabolic-related immune diseases.

Geniposide promotes splenic Treg differentiation to alleviate colonic inflammation and intestinal barrier injury in ulcerative colitis mice.

Geniposide has been proven to have a therapeutic effect on ulcerative colitis (UC) in animals, but its potential mechanism in UC remains to be clarified. The purpose of this study was to confirm the efficacy of geniposide in UC and to investigate the possible mechanism of geniposide in UC treatment. In vivo, geniposide relieved weight loss and reduced intestinal tissue damage in UC mice. Geniposide decreased the levels of IL-1ß and TNF-α and increased IL-10 levels in the colon and serum of UC mice. Geniposide increased FOXP3 expression in the colon and the number of CD4+ FOXP3+ cells in the spleen of UC mice. BD750 abolished the above regulatory effect of GE on UC mice. In vitro, geniposide increased the number of CD4+ FOXP3+ cells in spleen cells from normal mice, decreased the levels of IL-1ß, CCL2 and TNF-α in the supernatant of LPS-treated Caco-2 cells, and decreased the protein expression of Beclin-1 and Occludin in cacO-2 cells. Epirubicin inhibited the effect of geniposide on increasing the number of CD4+ FOXP3+ cells in spleen cells, attenuated the inhibitory effect of geniposide on proinflammatory factors and attenuated the upregulation of geniposide on tight junction proteins in LPS-treated Caco-2 cells in the coculture system. In conclusion, geniposide has an effective therapeutic effect on UC. Increasing Treg differentiation of spleen cells is the mechanism by which geniposide alleviates intestinal inflammation and barrier injury in UC.

Alisol B 23-acetate, a new promoter for cholesterol efflux from dendritic cells, alleviates dyslipidemia and inflammation in advanced atherosclerotic mice.

Atherosclerosis (AS) is characterized by dyslipidemia and chronic inflammation. In the high-fat environment, the lipid metabolism of dendritic cells (DCs) is abnormal, which leads to abnormal immune function, promotes the occurrence of immune inflammatory reactions, and promotes the development of AS. Alisol B 23-acetate (23B) is a triterpenoid in the rhizomes of Alisma, which is a traditional Chinese medicine. Here, we identified cholesterol metabolism-related targets of 23B through a virtual screen, and further transcriptome analysis revealed that 23B can change antigen presentation and cholesterol metabolism pathways in cholesterol-loaded DCs. In vitro experiments confirmed that 23B promoted cholesterol efflux from ApoE-/- DCs, reduced the expression of MHC II, CD80, and CD86, and inhibited the activation of CD4+ T cells and the production of inflammatory cytokines IL-12 and IFN-γ. In advanced AS mice, 23B can decrease triacylglycerol (TG) levels and increase high-density lipoprotein-cholesterol (HDL-C) levels in plasma and the expression of cholesterol efflux genes in the aorta. Neither helper T cells 1 (Th1) nor regulatory T cells (Tregs) in peripheral blood changed significantly in the presence of 23B, but 23B reduced the levels of IL-12 and IFN-γ in serum. However, 23B did not change the total cholesterol (TC) and low-density lipoprotein-cholesterol (LDL-C) levels in serum or lipid accumulation in the aorta. Moreover, 23B did not increase the production of IL-10 and TGF-ß1 in vivo or in vitro. These results indicate that 23B promotes cholesterol efflux from DCs, which can improve the immune inflammatory response and contribute to controlling the inflammatory status of AS.

Matrix Metalloproteinase-3 induces proteoglycan degradation in gouty arthritis model.

BACKGROUND: Gout is an inflammatory arthritis resulting from precipitation of monosodium urate (MSU) crystals in joints and surrounding tissues. However, the mechanism underlying high levels of uric acid inducing gouty arthritis has not been clarified. OBJECTIVE: The purpose was to investigate the role of Matrix Metalloproteinase-3 (MMP-3) in the development of gouty arthritis from hyperuricemia. METHOD: MSU crystal-induced gouty arthritis model and chondrocytes were used to evaluate changes of MMP-3 levels. Western blot, qPCR and ELISA were performed to detect MMP-3, Tissue Inhibitors of Metalloproteinase-1 (TIMP-1) and A Disintegrin and Metalloproteinase with Thrombospondin Motifs-4 (ADAMTS-4) expressions in rabbit chondrocytes. Expression of proteoglycan was determined through toluidine blue staining. Concentrations of glycosaminoglycan, Interleukin-6 (IL-6), Interleukin-1ß (IL-1ß) and Tumor Necrosis Factor-α (TNF-α) in chondrocytes were assessed via ELISA kits. Concentration of uric acid in supernate was tested by Automatic Analyzer. RESULTS: MMP-3 was significantly increased in rat serum, synovial fluid, cartilages and chondrocytes treated with high-level uric acid. Increased concentration of glycosaminoglycancould be observed in chondrocytes incubated with MMP-3, as well as the remarkable downregulation of proteoglycan expression. Furthermore, high-level uric acid contributed to the degradation of proteoglycan via the activation of MMP-3. IL-6, IL-1ß and TNF-α concentrations were increased significantly in 35 °C compared to 37 °C with MMP-3 and high-level uric acid. CONCLUSION: Our study showed that MMP-3 was enhanced by high levels of uric acid, which promoted proteoglycan degradation, and induced MSU crystallization in turn. A low temperature environment is an important factor in the development of gout.

Systematic Review With Meta-Analysis: Diagnostic, Prognostic and Clinicopathological Significance of CircRNA Expression in Renal Cancer.

BACKGROUND: Renal cancer (RC) is one of the most common malignant tumors of the urinary system, and molecular targets for the specific diagnosis and treatment of RC have been widely explored. The purpose of this study was to systematically analyze circular RNAs (circRNAs), which may serve as novel tumor markers in terms of the diagnosis, prognosis and clinicopathological characteristics of RC. METHODS: PubMed and Web of Science were systematically searched for literature as up to July 30, 2021. All included studies were evaluated by the evaluation system, and the results were satisfactory. Hazard ratios (HRs) and odds ratios (ORs) were used to assess the association of circRNAs with diagnostic and clinicopathological indicators. The sensitivity (SEN), specificity (SPE), positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio and area under the summary receiver operating characteristic curve (AUC) were combined to evaluate the diagnostic performance of circRNAs in RC. RESULTS: We included 22 studies that met the criteria, including 18 that were prognostic, 4 that were diagnostic, and 12 that were clinicopathologically relevant. In terms of prognosis, we found that upregulated circRNAs were positively associated with poor overall survival in patients with RC (HR=1.63, 95% CI=1.43-1.85). In terms of diagnosis, the combined SEN, SPE and AUC of circRNAs in the diagnosis of RC were 0.82, 0.84 and 0.89 (0.86-0.91), respectively. In terms of clinicopathological features, upregulated circRNAs were associated with the Fuhrman grade (OR=0.641, 95% CI=0.471-0.873), T stage (OR=0.236, 95% CI=0.141-0.396), TNM stage (OR=0.225, 95% CI=0.158-0.321) and lymphatic metastasis (OR=0.329, 95% CI=0.193-0.560). CONCLUSION: Our meta-analysis confirms that circRNAs may be candidate biomarkers for the diagnosis, prognosis, and clinicopathological indicators of RC.

Resveratrol ameliorates atherosclerosis induced by high-fat diet and LPS in ApoE-/- mice and inhibits the activation of CD4+ T cells.

BACKGROUND: Atherosclerosis (AS), which characterized with the accumulation of lipids on the vessel wall, is the pathological basis of many cardiovascular diseases (CVD) and seriously threatens human health. Resveratrol (RES) has been reported to be benefit for AS treatment. This research aimed to observe the effects of RES on AS induced by high-fat diet (HFD) and LPS in ApoE-/- mice and investigate the underlying mechanism. METHODS: ApoE-/- mice were fed with HFD companied with LPS to induce AS and RES was administrated for 20 weeks. Splenic CD4+ T cells were cultured and treated with anti-CD3/CD28 together with LPS, and RES was added. Serum lipids and the atherosclerotic areas of aortas were detected. The activation of CD4+ T cells were investigated both in vivo and in vitro and the expression of DNA methyltransferases (Dnmt) in CD4+ T cells were measured. RESULTS: In vivo, administration of RES prevented HFD and LPS induced dysfunction of serum lipids including TC (total cholesterol), TG (triglyceride), LDL-C (low density lipoprotein cholesterol) and HDL-C (high density lipoprotein cholesterol), ameliorated the thickened coronary artery wall and decreased the areas of atherosclerotic lesion on aortas. Besides, RES decreased the number of CD4+ T cells in peripheral blood, decreased the expression of CD25 and CD44, but not affected the expression of L-selectin (CD62L). In vitro, RES decreased the expression of Ki67, CD25 and CD44 in CD4+ T cells. Moreover, RES increased the secretion of IL-2, IL-10 and TGF-ß1, decreased IL-6. In addition, RES decreased both the mRNA and protein level of Dnmt1 and Dnmt3b in CD4+ T cells. CONCLUSION: These results indicated that RES ameliorated AS induced by HFD companied with LPS in ApoE-/- mice, inhibited the proliferation and activation of CD4+ T cells and regulated the expression of Dnmt1 and Dnmt3b.

Regulatory T Cell Plasticity and Stability and Autoimmune Diseases.

CD4+CD25+ regulatory T cells (Tregs) are a class of CD4+ T cells with immunosuppressive functions that play a critical role in maintaining immune homeostasis. However, in certain disease settings, Tregs demonstrate plastic differentiation, and the stability of these Tregs, which is characterized by the stable expression or protective epigenetic modifications of the transcription factor Foxp3, becomes abnormal. Plastic Tregs have some features of helper T (Th) cells, such as the secretion of Th-related cytokines and the expression of specific transcription factors in Th cells, but also still retain the expression of Foxp3, a feature of Tregs. Although such Th-like Tregs can secrete pro-inflammatory cytokines, they still possess a strong ability to inhibit specific Th cell responses. Therefore, the plastic differentiation of Tregs not only increases the complexity of the immune circumstances under pathological conditions, especially autoimmune diseases, but also shows an association with changes in the stability of Tregs. The plastic differentiation and stability change of Tregs play vital roles in the progression of diseases. This review focuses on the phenotypic characteristics, functions, and formation conditions of several plastic Tregs and also summarizes the changes of Treg stability and their effects on inhibitory function. Additionally, the effects of Treg plasticity and stability on disease prognosis for several autoimmune diseases were also investigated in order to better understand the relationship between Tregs and autoimmune diseases.

Yangyin Qingre Huoxue Method in Traditional Chinese Medicine Ameliorates Atherosclerosis in ApoE-/- Mice Suffering from High-Fat Diet and HSP65 Aggression.

Atherosclerosis (AS) is a complicated arterial disease resulting from abnormal lipid deposition and inflammatory injury, which is attributed to Yin deficiency, accumulation of heat materials, and stasis of blood flow in Traditional Chinese Medicine (TCM) theory. Thus, according to TCM theory, the method of nourishing Yin (Yangyin), clearing away heat (Qingre), and promoting blood circulation (Huoxue) is a reasonable strategy, which has achieved remarkable clinical efficacy in the treatment of AS, but the mechanisms remain to be known. In this study, we evaluated the effects of Yangyin Qingre Huoxue Prescription (YQHP) on AS in ApoE-/- mice suffering from a high-fat diet and heat shock protein (HSP65) attack. YQHP regulated levels of blood lipids and inflammation-linked cytokines as well as Th17/Treg ratio in peripheral blood. Suppressed IL-6-p-STAT3 signaling and restored IL-2-p-STAT5 signaling in the presence of YQHP may partake in the regulation of Th17 and Treg differentiation. Moreover, YQHP modulated transcriptional levels of costimulator CD80 in aortas as well corresponding to the downregulation of GM-CSF in serum and CD3 expression in CD4+ T cells, which might indicate the potential of YQHP to regulate antigen presenting cells. All these effects eventually promoted the improvement of atherosclerotic lesions. In addition, YQHP promoted less monocyte infiltration in the liver and lower levels of AST, ALT, and AKP production than simvastatin. Conclusively, lipid-regulating and anti-inflammatory functions mediated by YQHP with lower hepatotoxicity than simvastatin hindered the progression of HSP65 aggravated AS in ApoE-/- mice, indicating the effectiveness of Yangyin Qingre Huoxue Method in the treatment of AS.

MicroRNA­143­3p contributes to the regulation of pain responses in collagen­induced arthritis.

Patients with rheumatoid arthritis (RA) suffer from pain, which is associated with inflammation, peripheral and central pain processing, and joint structure damage. The aim of the present study was to investigate a key microRNA (miR) and its target genes that are involved in the pain responses of RA, and to clarify the mechanism of pain regulation. Collagen­induced arthritis (CIA) was induced in DBA/1 and C57BL/6 mice. The paw swelling, mechanical withdrawal threshold (MWT), thermal withdrawal latency (TWL), and expression levels of tumor necrosis factor (TNF)­α and prostaglandin (PG)E2 in the sera were investigated. Decreased MWT and TWL, and increased TNF­α and PGE2, in the CIA model group were observed in DBA/1 and C57BL/6 mice. DBA/1 mice exhibited greater hyperalgesia and higher levels of inflammatory mediators. miR­143­3p expression in the blood and the dorsal root ganglion (DRG) were detected, and low miR­143­3p expression was demonstrated in the blood and DRG tissue of CIA mice. The target genes of miR­143 were predicted and analyzed. A total of 1,305 genes were predicted and 55 pain­associated genes were obtained. Prostaglandin­endoperoxide synthase 2 (Ptgs2), MAS related GPR family member E (Mrgpre), prostaglandin D2 receptor and Tnf were selected as target genes of miR­143. DRG cells were cultured and transfected with miR­143­3p inhibitor or mimic. The expression of Mrgpre, Ptgs2 and Tnf was significantly inhibited following miR­143­3p mimic transfection, while the expression of Mrgpre, Ptgs2 and Tnf was increased following inhibitor transfection. Additionally, the expression of pain­associated genes in the DRG of mice was investigated and the expression of Ptgs2, Mrgpre and Tnf in the DRG of CIA mice was also significantly upregulated. These results revealed that CIA mice exhibited marked hyperalgesia and high levels of inflammatory pain mediators. Low expression of miR­143­3p negatively regulated the pain­associated target genes, including Mrgpre, Ptgs2 and Tnf, thereby affecting chronic inflammatory pain and neuropathic pain in RA.

Hepatitis B, C, and D virus infection showing distinct patterns between injection drug users and the general population.

BACKGROUND AND AIM: Hepatitis B, C, and D virus (HBV, HCV, and HDV) infections are known to be prevalent in injection drug users (IDUs); however, the relationship between the molecular epidemiologic features of hepatitis virus infection in high-risk individuals and the general population has not yet been established. METHODS: In total, 1049 IDUs and 672 individuals who underwent physical examinations at Chenzhou hospital, Hunan Province, China, were enrolled. HBV, HCV, and HDV infections were screened with serologic tests in both populations. HBsAg-positive, anti-HCV IgG-positive, and anti-HDV IgG-positive samples were further confirmed by polymerase chain reaction, quantitative polymerase chain reaction, and DNA sequencing. RESULTS: Significantly higher HBV (21.54 vs 16.52%, P = 0.01), HCV (45.95% vs 1.34%, P < 0.001), and HDV (5.62% vs 0.30%, P < 0.001) infections were detected in IDUs compared with the general population. The dual infection of HBV/HCV or HBV/HDV was also significantly higher in IDUs than in the general population. HBV genotype B and HDV genotype II were dominants in both populations. HCV infection showed genotype 6a (49.52%) dominant in IDUs, but genotype 1b accounted for 50% infection, which was followed by genotype 6a (33.33%) in the general population. Higher viral loads were associated with HBV genotype B and HCV genotype 6a compared with non-dominant genotypic infections. CONCLUSIONS: HBV and HDV infections shared similar patterns by IDUs and the general populations, and HCV infection exhibited distinct features between two populations. Our results suggest different molecular epidemiologic characteristics of HBV, HCV, and HDV infection in two populations.

Alantolactone exhibited anti-herpes simplex virus 1 (HSV-1) action in vitro.

The aim of this study was to determine the inhibitory action of alantolactone, a gradient of traditional Chinese medicine Inulae Radix (Tu-Mu-Xiang), on herpes simplex virus 1 (HSV-1). African green monkey kidney cells (Vero cells) were infected with HSV-1 and the protective effects of alantolactone on Vero cells were examined. At concentrations of 10(-6), 10(-7), and 10(-8) g/mL, alantolactone did not have a marked harmful effect on the viability of Vero cells according to an MTT assay. Based on the cytopathic effect (CPE) and MTT assays, alantolactone at these concentrations exhibited antiviral action and protected cells from being damaged by HSV-1. Results indicated that alantolactone had potent anti-HSV-1 action and provided evidence for use of Inulae Radix in the treatment of HSV-1 infection.

The forecast of anticancer targets of cryptotanshinone based on reverse pharmacophore-based screening technology.

Anticancer targets of cryptotanshinone were evaluated and rapidly forecasted with PharmMapper, a reverse pharmacophore-based screening platform, as well as drug target databases, including PDTD, DrugBank and TTD. The pathway analyses for the collection of anticancer targets screened were carried out based on the KEGG pathway database, followed by the forecast of potential pharmacological activities and pathways of the effects of cryptotanshinone, and verification of some of the targets screened using whole cell tests. The results showed that a total of eight targets with anticancer potential were screened, including MAP2K1, RARα, RXRα, PDK1, CHK1, AR, Ang-1 R, and Kif11. These targets are mainly related to four aspects of the cancer growth: the cell cycle, angiogenesis, apoptosis, and androgen receptor. The cell tests showed that cryptotanshinone can inhibit the viability of human hepatoma cells SMMC-7721, which is related to the reduction of expression of MAP2K1 mRNA. This method provides a strong clue for the study of the anticancer effects and mechanisms of action of cryptotanshinone in the future.

Shikonin reduces endometriosis by inhibiting RANTES secretion and mononuclear macrophage chemotaxis.

Endometriosis is a common disease in females of reproductive age and has the classic characteristic of mononuclear cell infiltration into lesions. Shikonin is an anti-inflammatory phytocompound obtained from Lithospermum erythrorhizon whose potential therapeutic effects in the treatment of endometriosis remain unclear. The working hypothesis of the present study was that shikonin is capable of inhibiting the development of endometriosis by inhibiting the chemotactic effect. In a murine model of endometriosis, shikonin significantly inhibited the growth of human endometrial tissue implanted into severe combined immunodeficiency (SCID) mice (P<0.05) and no adverse effects were observed. Mouse regulated upon activation normal T-cell expressed and secreted (mRANTES) levels in the peritoneal fluid of the animal endometriosis model were higher than those in normal SCID mice (P<0.05) and decreased significantly following shikonin treatment in a dose-dependent manner (P<0.05). Peritoneal fluid from SCID mice treated with shikonin inhibited the chemotaxis of monocytes; this inhibitory effect was eradicated by mRANTES antibody. In vitro, shikonin significantly inhibited RANTES expression in U937 cells that were cultured alone or co-cultured with human mesothelial and endometrial stromal cells. In addition, shikonin inhibited the RANTES-induced chemotaxis of U937 cells (P<0.05). The results indicate that shikonin inhibits the development of endometriosis by various mechanisms, including the inhibition of RANTES expression and the reduction of mononuclear cell migration to lesions. Therefore, shikonin may be a novel, useful and safe agent for treating endometriosis.

Nasal immunization with different forms of heat shock protein-65 reduced high-cholesterol-diet-driven rabbit atherosclerosis.

Heat shock protein-65 (Hsp65) is an important pro-atherogenic factor, but nasal immunization of Hsp65 can induce immune tolerance and reduce atherosclerotic inflammation. Here, we describe the effects of different forms of Hsp65 antigen inoculated, i.e. Hsp65 DNA, Hsp65 protein, Hsp65 DNA prime combined with Hsp65 protein boost, on high-cholesterol-diet-driven rabbit atherosclerosis lesions. The results showed that sera anti-Hsp65 IgG antibodies were lower induced and protective anti-Hsp65 IgA in nasal cavity and lung were induced after rabbits were immunized with different forms of Hsp65. However, the avidity between Hsp65 IgG antibody and antigen is different. Hsp65 DNA prime-protein boost could further improve the avidity compared with Hsp65 DNA immunization alone, but only immunization with Hsp65 protein alone obtain the best avidity. In addition, more protective IL-10 and less adverse IFN-γ were produced and sera TC and LDL-C were decreased obviously at different extents in immunized groups. As a result, atherosclerosis lesions were significantly attenuated, but only nasal immunization with Hsp65 protein alone may be the best effective form of Hsp65 antigen to control atherosclerotic lesions. The results demonstrated that different forms of Hsp65 antigen could produce different effects in the treatment of atherosclerosis. Immunization mucosally with Hsp65 protein could be a promising therapeutic method for atherosclerosis.

Regulatory effect of melatonin on cytokine disturbances in the pristane-induced lupus mice.

Systemic lupus erythematosus (SLE) develops in relation to many environmental factors. In our opinion, it is more important to investigate the effect of melatonin on the environmental- related SLE. In the present study, 0.5 ml pristane were used to induce SLE in female BALB/c mice. Melatonin (0.01, 0.1, 1.0 mg/kg) was orally administered immediately after pristane-injection for 24 weeks. IgM anti ssDNA and histone antibodies were detected after 0, 1, 2, 4, 8 weeks pristane injection. The levels of IL-2, IL-6 and IL-13 were detected after 24 weeks. Renal lesions were also observed. The results showed that melatonin antagonized the increasing levels of IgM anti ssDNA and histone autoantibodies. Melatonin could also decrease the IL-6 and IL-13 production and increase the IL-2 production. Besides, melatonin could lessen the renal lesions caused by pristane. These results suggested that melatonin has a beneficial effect on pristane-induced lupus through regulating the cytokines disturbances.