Effects of polyphyllin Ⅵ on the proliferation and apoptosis of glioma cells and potential mechanism / 中国药房

OBJECTIVE To investigate the effects of polyphyllin Ⅵ（PPⅥ） on the proliferation and apoptosis of glioma cells and potential mechanism. METHODS Using human glioma LN229 cells as objects， MTT assay was used to detect the survival rate after treated with different concentrations of PPⅥ [0 （control group）， 1， 2， 4， 8， 16， 32， 64 μmol/L] for different time （24， 48， 72 h）. The clone formation experiments were adopted to detect the number of cell clones and clone formation rate after being treated with different concentrations of PPⅥ [0 （control group）， 2， 4， 8 μmol/L] for 14 days. The flow cytometry and Western blot assay were used to detect the apoptotic rate of cells， the expressions of apoptosis-related protein [B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， cleaved caspase-3]， and the expressions of related proteins of Fas/Fas ligand （FasL） death receptor pathway and protein kinase B （Akt）/glycogen synthesis kinase-3β （GSK-3β） pathway after being treated with different concentrations of PPⅥ [0（control group）， 4， 8 μmol/L] for 24 h. RESULTS Compared with the control group， the survival rate of cells， the number of clones and clone formation rate， the protein expression of Bcl-2， and the phosphorylation levels of Akt and GSK-3β protein were decreased significantly in different concentration groups of PPⅥ （P＜0.05 or P＜0.01）. The apoptotic rate， the protein expressions of Bax， cleaved caspase-3， Fas， FasL and cleaved caspase-8 were increased significantly （P＜0.05 or P＜ 0.01）. CONCLUSIONS PPⅥ can inhibit the proliferation and induce the apoptosis of human glioma LN229 cells， which may be related to the activation of the Fas/FasL death receptor pathway and the inhibition of the Akt/GSK-3β pathway.

Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica / 口腔疾病防治

Objective@# To investigate the transcriptomic changes in primary human oral keratinocytes (pHOKs) after coculture with Prevotella melaninogenica (P.m) and to verify the changes in human oral keratinocyte (HOK) cell lines.@*Methods@# pHOK was isolated and cocultured with P.m for 0, 4 and 24 h. Total RNA was extracted, a gene library was constructed, transcriptional sequencing was performed, differentially expressed genes (DEGs) were analyzed, gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and the validation of DEGs was performed by qRT-PCR and Western Blot in the HOK and P.m coculture cell model. @*Results @#1 788 DEGs were detected between the 4 h group and control group, including upregulated DEGs such as lymphocyte cytosolic protein 1(LCP1), keratin 7 (KRT7) and Cilia and flagella associated protein 251(CFAP251) and downregulated DEGs such as FERM, ARH/RhoGEF and Pleckstrin domain protein 1 (FARP1), WW domain containing transcription regulator 1(WWTR1) and Discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2). 1 832 DEGs were detected between the 24 h group and control group, including upregulated DEGs such as LCP1, complement C1s(C1S), kynureninase (KYNU) and downregulated DEGs such as phosphoserine aminotransferase 1 (PSAT1), FARP1 and FKBP prolyl isomerase 10 (FKBP10). There were 1 090 common differentially expressed genes (cDEGs) in the 4 h and 24 h groups, including LCP1, KYNU and long intergenic nonprotein coding RNA 958 (LINC00958). The GO pathways were mainly enriched in response to lipopolysaccharide and the molecules of bacterial origin and apical part of the cell. KEGG pathway analysis revealed enrichment in the interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Toll-like receptor (TLR) pathway, etc. We verified the expression of a cDEG, Myosin1B (MYO1B), and qRT-PCR and Western Blot analysis showed that MYO1B expression was significantly upregulated between the control group and the P.m cocultured group (P<0.001), and its expression followed a time-dependent and concentration-dependent manner. @*Conclusion@#P.m played an important role in the transcriptome of oral keratinocytes.

Study on long noncoding RNA LBX2-AS1 regulates glioma cell proliferation, migration and apoptosis through epidermal growth factor receptor signaling pathway / 中国医师进修杂志

Objective:To investigate the mechanism of long noncoding RNA (lncRNA) LBX2-AS1 regulating glioma cell proliferation, migration and apoptosis through epidermal growth factor receptor (EGFR) signaling pathway.Methods:From April 2018 to August 2021, glioma U251 cells (U251 cells for short) were divided into control group and observation group, with 4 strains in each group. The control group was routinely cultured, and the observation group was transfected with specific small interfering RNA (siRNA) targeting LBX2-AS1. The proliferation ability of U251 cells was detected by methyl thiazol tetrazolium method, the metastasis rate of U251 cells was detected by scratch test, the apoptosis rate of U251 cells was detected by flow cytometry, and the expression of total protein and vascular endothelial growth factor (VEGF), phosphorylated inositol 3 kinase (p-PI3K), phosphorylated protein kinase B (p-Akt), phosphorylated Ras (p-Ras) and phosphorylated Raf (p-Raf) protein were detected by Western blot.Results:The proliferation ability and metastasis rate of U251 cells in observation group were significantly lower than those in control group: (27.15 ± 1.38)% vs. (63.54 ± 2.47)% and (37.09 ± 3.74)% vs. (82.17 ± 9.24)%, the apoptosis rate of U251 cells was significantly higher than that in control group: (69.17 ± 5.83)% vs. (17.58 ± 1.22)%, and there were statistical differences ( P<0.01). The expression of total protein and VEGF, p-PI3K, p-Akt, p-Ras, p-Raf protein of U251 cells in observation group were significantly lower than those in control group (1.52 ± 0.23 vs. 2.39 ± 0.31, 0.73 ± 0.08 vs. 1.68 ± 0.45, 0.57 ± 0.11 vs. 1.89 ± 0.31, 0.68 ± 0.06 vs. 1.74 ± 0.51, 0.84 ± 0.12 vs. 1.99 ± 0.63 and 0.71 ± 0.08 vs. 1.52 ± 0.37), and there were statistical differences ( P<0.01). Conclusions:The lncRNA LBX2-AS1 is highly expressed in glioma cells. Silencing the expression of lncRNA LBX2-AS1 inhibits the proliferation and metastasis of glioma cells through EGFR pathway.

Active fractions of Camellia nitidissima inhibit non-small cell lung cancer via suppressing epidermal growth factor receptor / 中国中药杂志

The present study explored the effects and its underlying mechanisms of four active fractions of Camellia nitidissima(leaf polyphenols, leaf saponins, flower polyphenols, and flower saponins in C. nitidissima) in inhibiting the proliferation and migration of non-small cell lung cancer(NSCLC) by suppressing the epidermal growth factor receptor(EGFR). MTT assay was used to detect the effect of four active fractions on the proliferation of NCI-H1975 and HCC827 cells. Wound healing assay and Transwell assay were adopted to evaluate the effect of four active fractions on the migration of NSCLC. The effect of four active fractions on the enzyme activity of EGFR was detected. Molecular docking was carried out to explore the direct action capacity and action sites between representative components of the four active fractions and EGPR. Western blot assay was employed to investigate the effect of four active fractions on the protein expression in EGFR downstream signaling pathways. The results of the MTT assay indicated that the cell viability of NCI-H1975 and HCC827 cells was significantly inhibited by four active fractions at 50, 100, 150, and 200 μg·mL~(-1) in a dose-dependent manner. Wound healing assay and Transwell assay revealed that the migration of NCI-H1975 and HCC827 cells was significantly suppressed by four active fractions. In addition, the results of the protein activity assay showed that the enzyme activity of EGFR was significantly inhibited by four active fractions. The molecular docking results confirmed that various components in four active fractions possessed strong binding activity to EGFR enzymes. Western blot assay revealed that four active fractions down-regulated the protein expression of EGFR and its downstream signaling pathways. It is concluded that the four active fractions of C. nitidissima can inhibit NSCLC. The mechanism may be related to EGFR and its downstream signaling pathways. This study provides a new scientific basis for the clinical treatment of NSCLC with active fractions of C. nitidissima, which is of reference significance for further research on the anti-tumor mechanism of C. nitidissima.

Impact of immune inflammation on the proliferation and apoptosis of benign prostatic hyperplasia cells / 中华男科学杂志

Objective@#To investigate the impact of macrophage-induced immune inflammation on the proliferation and apoptosis of BPH cells and its possible mechanism.@*METHODS@#Macrophages were stimulated with phorbol myristate acetate, co-cultured with BPH-1 cells, and then treated with the androgen receptor (AR) inhibitor or anti-CD40L antibody. The immunohistochemical biomarkers of the T lymphocytes (CD4 and CD8), B lymphocyte (CD20) and macrophages (CD68), AR, CD40/CD40L, and inflammatory factors IL-1, IL-6 and TNF-α were measured before and after treatment. The proliferation and apoptosis of the cells were observed by MTT assay, colony-forming assay and flow cytometry, and the expressions of cell apoptosis- and MAPK signaling pathway-related proteins were determined by qRT-PCR and Western blot.@*RESULTS@#Significantly increased proliferation and decreased apoptosis of the cells, up-regulated expressions of Bcl-2, IL-1, IL-6, TNF-α, AR, CD40 and CD40L, and down-regulated expression of Bax were observed in the BPH-1 cells co-cultured with macrophages (the M-BPH-1 group) compared with those in the blank control (B-BPH-1) group (P < 0.01). In comparison with the BPH-1 cells treated with normal saline, those treated with either low-dose CD40L (L-CD40L) or high-dose CD40L (H-CD40L) showed markedly inhibited proliferation, increased apoptosis, up-regulated expression of Bax, and down-regulated expressions of Bcl-2, IL-1, IL-6 and TNF-α (P < 0.01), and those in the low- and high-dose AR (L-AR and H-AR) inhibitor groups exhibited remarkably reduced proliferation, increased apoptosis, down-regulated expressions of Bcl-2, IL-1, IL-6 and TNF-α, and up-regulated expression of Bax (P < 0.01). The phosphorylation levels of JNK, ERK and P38 were significantly elevated in the M-BPH-1 group, but declined in the H-CD40L and the H-AR inhibitor groups compared with those in the B-BPH-1 group, all in a concentration-dependent manner (P < 0.01).@*CONCLUSIONS@#Macrophage-induced immune inflammation regulates AR and CD40/CD40L expressions and promotes the proliferation and inhibits the apoptosis of BPH-1 cells by activating the MAPK signaling pathway. /.

Complement 3-complement 3a receptor pathway and neurodegenerative diseases / 中华神经医学杂志

Neurodegenerative disease is a kind of degenerative diseases of the central nervous system that seriously endanger human health. The complement 3 (C3)-complement 3a receptor (C3aR) pathway is one of the important pathways for classical complement cascade activation. A large number of studies have shown that the C3-C3aR pathway can mediate and regulate the interaction of astrocyte-microglia axis in neurons, resulting in function changes in central nervous system. In addition, studies in recent years have found that the C3-C3aR pathway is closely related to the occurrence and progress of neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, stroke, and epilepsy. This article reviews the progress of C3-C3aR pathway and discuss the role of C3-C3aR pathway in several important neurodegenerative diseases, and it provides a new idea fo treatment of these diseases.

Protective effect of Lycium barbarum polysaccharide on reproductive system injury of female rats induced by 2,4-dichlorophenoxyacetic acid / 环境与职业医学

Background 2,4-Dichlorophenoxyacetic acid (2,4-D) is widely used as a broad-leaved herbicide and plant growth regulator. Related studies have shown that 2,4-D has neurotoxicity, ability to disrupt endocrine function, genotoxicity, carcinogenicity, and reproductive toxicity. Objective This experiment is conducted to investigate the effect of 2,4-D exposure on reproductive system of female rats, and to preliminarily explore the potential ameliorative effect of Lycium barbarum polysaccharide (LBP) and its possible mechanism. Methods Twenty-four SPF female SD rats with six rats in each group were randomly divided into a blank control group (deionized water 1 mL·d−1), an exposure group (75 mg·kg−1 2,4-D), an LBP control group (50 mg·kg−1 LBP), and an LBP intervention group (75 mg·kg−1 2,4-D + 50 mg·kg−1 LBP). The rats were given intragastric administration once a day for 28 consecutive days. Body weight was measured every two days. After exposure, ovary and uterus were weighed and organ coefficients were calculated; the pathological changes of ovary and uterus were detected by hematoxylin-eosin staining (HE); the level of estradiol (E2) in serum was detected by ELISA; the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) in serum were measured by corresponding kits; the apoptosis of ovarian and uterine cells was detected by TUNEL fluorescence staining; and the protein expression levels of Fas, FasL, FADD, Pro-Caspase-8, Cleaved-Caspase-8, Pro-Caspase-3, and Cleaved-Caspase-3 in ovarian tissues were detected by Western blotting. Results Compared with the blank control group, the ovarian structure of the exposure group was abnormal, the number of follicles at different developmental stages decreased, morphological changes were observed, and the number of atretic follicles increased; the endometrium was incomplete, with different degrees of nuclear pseudostratification and decreased number of glands in lamina propria. Compared with the exposure group, the ovarian structure of the LBP intervention group was complete, and the follicles at different developmental stages increased in amount, remained intact, and were arranged closely; the uterine structure was relatively intact, showing decreased endometrial loss and nuclear pseudostratification. There were significant differences in the levels of SOD, GSH-Px, E2, and MDA among the four groups (F=86.1, 26.2, 43.3, and 22.3, all P<0.01). Compared with the blank control group, the levels of serum SOD, GSH-Px, and E2 decreased in the exposure group (P<0.01), while the concentration of MDA increased (P<0.01). Compared with the exposure group, the levels of serum SOD, GSH-Px, and E2 in the LBP intervention group increased (P<0.01), and the concentration of MDA decreased (P<0.01). There were significant differences in the apoptosis rates of ovarian and uterine cells among the four groups (F=64.8, 55.5, both P<0.01). Compared with the blank control group, the apoptosis rates of ovarian and uterine cells increased in the exposure group (P<0.01). Compared with the exposure group, the apoptosis rates of ovarian and uterine cells decreased in the LBP intervention group (P<0.01). There were significant differences in the expression levels of death receptor pathway-related proteins in ovarian tissues among the four groups (all P<0.05). Compared with the blank control group, the expression levels of Fas, FasL, FADD, Pro-Caspase-8, Cleaved-Caspase-8, Pro-caspase-3, and Cleaved-Caspase-3 were increased in the exposure group (P<0.05 or 0.01). Compared with the exposure group, the expression levels of above proteins were decreased in the LBP intervention group (P<0.05 or 0.01). Conclusion The study findings reveal that 2,4-D can induce oxidative stress and further mediate Fas-FasL pathway to induce apoptosis, resulting in reproductive system damage in female rats. LBP can reduce the oxidative stress level, down-regulate the expression of Fas-FasL pathway-related proteins, and reduce the apoptosis of germ cells, therefore protecting reproductive system of female rats.

Inflammatory pathways and Toll-like receptors related protein expression on acute gouty arthritis model rats / 中华风湿病学杂志

Objective:Sodium urate was used to induce acute gouty arthritis rat model, and to observe the inflammatory response of rats and the intervention effect of diclofenac sodium on the expression of Toll-like receptor-related (TLR) protein of ankle joint.Methods:Thirty males specific pathogen free (SPF) grade Wistar rats were used to develop the models. Random number table method was used to divide the rats into normal saline control group, model group, and drug group (diclofenac sodium t 1.35 mg/g body weight), 10 rats in each group. After fully grinding the sodium urate crystals, an appropriate amount of saline and Tween-80 (9∶1) was added to make a suspension, and the sodium urate crystals (25 mg/ml) were injected to the right posterior ankle of the rats in the model and drug groups. The solution was 0.2 ml, and rats in the sham group were injected with 0.2 ml of normal saline at the same location. After the model was established, drug and equal volume of purified water were administrated intragastrically once a day for 7 days. The toe volume device was used to measure the joint swelling of the rat (at 4 h, 8 h, 24 h, 48 h, 72 h) , and blood was taken from the abdominal aorta after anesthesia to determine the rat kidney function, interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) content, the rat ankle joint TLR4, myeloid differentiation factor (MyD88), NF-κBp65 protein expression were determined using Western blot and immunohistochemical methods. Multiple comparisons were carried out using single factor analysis of variance (ANOVA), comparing the two groups by using LSD- t, the comparison of different times using repetitive measure analysis of variance (repeated measures). Results:After the models were established, the rat's right ankle joint showed various degrees of redness, slow walking, and unresponsiveness. Compared with the normal saline control group, under the light microscope, the ankle synovial cells of the model group proliferated, with localized degeneration and necrosis, and many inflammatory cell infiltration. The rat serum inflammatory factors IL-1β, IL-6, TNF-α in the diclofenac sodium group [(24.6±3.3) pg/ml, (151±21) pg/ml, (61±16) pg/ml] were significantly reduce compared with model group [(28.4±4.3) pg/ml, (173±26) pg/ml, (81±5) pg/ml] ( t=2.296, P<0.01; t=2.909, P<0.01; t=2.352, P<0.01). Compared with normal saline group, variance analysis showed that the NF-κBp65, MyD88, TLR4 protein expression of ankle joint detected by Western bolt method and immunohistochemistry method was significantly increased in the model group. Compared with the model group, diclofenac sodium the ankle tissue protein expression of NF-κBp65, MyD88, and TLR4 was significantly inhibited. There were statistical significances in three groups ( P<0.05 or P<0.001). Conclusion:The level of inflammatory factors in acute gout arthritis rats model induced by sodium urate crystals is increased, and the expression of TLR4/MyD88/NF-КBp65 proteins in ankle joint tissue is increased, which affects the TLR signaling pathway. Diclofenic has inhibitory and relieving effects.

Effect of Astragalus Polysaccharide on Sweet Taste Receptor Pathway in Intestine of Rat Model Induced by High-sugar and High-fat Diet / 中国实验方剂学杂志

Objective:To observe the effect of astragalus polysaccharide (APS) on taste receptor 1 member 2 (T1R2)/taste receptor 1 member 3 (T1R3) sweet taste receptor pathway in intestine of rat model induced by high-sugar and high-fat diet. Method:SD rats were randomly divided into normal group, high-sugar and high-fat group and astragalus polysaccharide group. Rats in high-sugar and high-fat group and astragalus polysaccharide groups were fed with high-sugar and high-fat diet for 16 weeks, while rats in astragalus polysaccharide group were fed with APS (0.7 g·kg-1, per day) for 8 weeks during this period. Serum samples were collected to determine the levels of fasting blood glucose, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). Intestinum tenue was collected to determine mRNA expressions of T1R2/T1R3, α-gustducin (Gα gust), transient receptor potential cation channel subfamily member 5 (TRPM5) and proglucagon (PG) gene by Real-time PCR, and protein expressions of T1R2, Gα gust and glucagon-like peptide-1 (GLP-1) protein by Western blot. Result:Rats in high-sugar and high-fat group had significantly higher levels of TC, TG and LDL-C, and lower HDL-C level in serum than those in normal group (Pα gust and PG genes in intestine, were significantly down-regulated in high-sugar and high-fat group (PPα gust, TRPM5 and PG genes in intestine were significantly up-regulated in astragalus polysaccharide group (Pα gust and GLP-1 protein expressions was consistent with that of T1R2, Gα gust and GLP-1 mRNA expressions. Protein expressions of T1R2, Gα gust and GLP-1 and mRNA expression of T1R3 were significantly lower in astragalus polysaccharide group than those of control group (PConclusion:APS could improve disturbance of lipid metabolism and impairment of intestinal sweet taste receptor pathway for rat model induced by high-sugar and high-fat diet.

Pathomechanisms of pericyte-myofibroblast transition in kidney and interventional effects of Chinese herbal medicine / 中国中药杂志

In the kidney, pericyte is the major source of myofibroblast (MyoF) in renal interstitium. It is reported that pericyte-myofibroblast transition（PMT）is one of the important pathomechanisms of renal interstitial fibrosis（RIF）. Among them, the main reasons for promoting RIF formation include pericyte recruitment, activation and isolation, as well as the lack of pericyte-derived erythropoietin. During the PMT startup process, pericyte activation and its separation from microvessels are controlled by multiple signal transduction pathways, such as transforming growth factor-β（TGF-β）pathway, vascular endothelial growth factor receptor (VEGFR) pathway and platelet derived growth factor receptor (PDGFR) pathway;Blocking of these signaling pathways can not only inhibit PMT, but also suppress renal capillaries reduction and further alleviate RIF. In clinic, many traditional Chinese medicine compound prescriptions, single traditional Chinese herbal medicine (CHM) and their extracts have the clear effects in alleviating RIF, and some of their intervention actions may be related to pericyte and its PMT. Therefore, the studies on PMT and its drug intervention will become the main development direction in the research field of anti-organ fibrosis by CHM.

Xanthotoxin induces apoptosis in SGC-7901 cells through death receptor pathway / 中草药·英文版

Objective: To investigate the effects of xanthotoxin from Apiaceae medicinal plants on cell proliferation and apoptosis, and explore its mechanism of action against human gastric carcinoma SGC-7901 cells in vitro. Methods: SGC-7901, HepG-2, MCF-7, and A549 cells were treated with different concentrations of xanthotoxin (10, 20, 60, 80, 100, 120, 140, and 160 µg/mL) for 48 h, and the cell viability (IC50) was determined by MTT assay; Xanthotoxin-induced apoptosis in cells was observed by using Hoechst 33258 Staining Kit and Annexin V-FITC Apoptosis Detection Kit; Flow cytometry was used to detect apoptosis related proteins of Fas/FasL, Bid, and DR5/TRAIL proteins in human gastric carcinoma SGC-7901 cells after being treated by xanthotoxin; The influence of xanthotoxin on Caspase-8 protein expression in the cells was determined by Flouormetric Assay Kit. Results: Xanthotoxin obviously inhibited SGC-7901, HepG-2, MCF-7, and A549 cells proliferation, and its inhibition was in a concentration-dependent manner; flow cytometry results showed that in a certain concentration range, xanthotoxin can increase the expression levels of Fas/FasL and DR5/TRAIL proteins in a concentration-dependence manner. The content of Bid protein in cells was increased, and it showed concentration-dependence. Conclusion: Xanthotoxin may induce SGC-7901 cells apoptosis in a certain concentration range through the Fas/FasL protein mediated death receptor pathway, or by DR5/TRAIL mediated death receptor pathway, and increase the expression level of death receptor protein, activation Caspase-8, activating downstream effect factor, inducing cell apoptosis, or activate Caspase-8 cutting activate protein Bid, and then enter the mitochondrial pathway, induction of apoptosis.

Effect and mechanism of astragaloside Ⅳ on Toll-like receptor pathway in fibrotic mice after renal ischemia-reperfusion / 中国中药杂志

The aim of this paper is to study the effect of astragaloside Ⅳ on renal fibrosis mice with ischemia-reperfusion injury (IRI) and discuss the mechanism. Male C57BL/6 50 mice were randomly divided into four groups, namely Sham-operated group, model group, AS-Ⅳ prevention group and AS-Ⅳ treatment group. Since the day of surgery, the mice in astragaloside Ⅳ prevention group were treated with astragaloside Ⅳ by gavage for 30 days at the dose of 30 mg·kg⁻¹·d⁻¹. At the 60th day after surgery, the mice in astragaloside Ⅳ treatment group were treated with astragaloside Ⅳ 100 by gavage for 30 days at the dose of 30 mg·kg⁻¹·d⁻¹. The mice in Sham-operated group and model group were treated with double distilled water containing 0.1% ethanol instead of astragaloside Ⅳ. Serum creatinine and blood urea nitrogen were detected by chemical methods. Histopathological changes and collagen deposition of affected kidneys were observed under optical microscope by HE and Masson staining. The expression levels of Toll like receptor pathway related molecules (TLR4,MyD88,TRAF6,TRAM,TRIF,NF-κB,TNF-α,IL-6, IFN-) in affected kidneys were observed by immunohistochemistry, Western blot methods and reverse transcription-PCR atprotein and mRNA levels in each group. The results showed that the degrees of fibrosis and histopathological damage of affected kidneys of mice in model group were the most obvious. And the expression levels of TLR4/MyD88 dependent signaling pathway-related molecules (TLR4 and MyD88, TRAF6 and NF-κB) in affected kidneys of mice in model group were the highest. At the same time, there was no difference in the expression levels of TLR4/MyD88 independent signaling pathway-related molecules（TRAM, TRIF）among sham-operated group, model group, astragaloside IV prevention group and astragaloside Ⅳ treatment group. In astragaloside Ⅳ prevention group and astragaloside Ⅳ treatment group, the injury of affected kidney was obviously reduced, and the protein expression levels of TLR4/MyD88 dependent signaling pathway-related molecules were also correspondingly reduced; at the same time, the expressions of terminal inflammatory cytokines (TNF-α,IL-6, IFN-) were suppressed. Therefore, astragaloside Ⅳ may improve renal interstitial fibrosis in mice after IRI by inhibiting the expression of TLR4/MyD88 dependent signaling pathway and the release of inflammatory cytokines (TNF-α,IL-6, IFN-), while the TLR4/MyD88 independent signaling pathway may not be involved in the process of renal fibrosis after ischemia-reperfusion injury.

Effects of Modified Danggui Beimu Kushen Pills on Toll-like Receptor Pathway Markers on MFC Gastric Cancer Mice / 中国中医药信息杂志

Objective To study the effects of modified Danggui Beimu Kushen Pills on the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissues on MFC gastric cancer bearing mice; To discuss relevant mechanism of action. Methods MFC gastric cancer bearing mice were employed to perform anti-tumor experiment in vivo in this study. A total of eligible 48 mice were randomly divided into model group, DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups, modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. The treatment was conducted once a day, and lasted for 14 continuous days. After the last administration of gavage orally treatment, all mice were anaesthetized and killed by cervical dislocation method to obtain tumor tissue completely for further HE staining measure and detection of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissue with the method of RT-qPCR and immunohistochemistry. Meanwhile, the tumor growth was observed and the general conditions of mice were recorded. Results The model group was rich in tumor cells; the sizes of cells were different; the volume was large; the nucleus was deeply stained and the heterotypic shape was obvious, and the small focal necrosis was seen. The number of tumor cells in each administration group was reduced; the arrangement was loose; the cell volume was significantly reduced, and the nuclear pyknosis was reduced. Cell necrosis significantly increased; the number of interstitial blood vessels decreased; collagen fibers increased, especially in modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. Compared with the model group, the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 mRNA and protein decreased in each administration group. TLR2, TLR4, TLR6, TRAF6 and MyD88 were lighter and weakly positive expressed in modified Danggui Beimu Kushen Pillshigh-dose and low-dose combined with DDP groups, the protein changes were more obvious Compared with DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups. Conclusion Modified Danggui Beimu Kushen Pills can down-regulate TLR2, TLR4, TLR6, TRAF6 and MyD88 expression of tumor tissue in MFC gastric cancer bearing mice at both mRNA and protein levels to play anti-tumor pharmacology action.

Research progress of B cell receptor pathway inhibitors in the treatment of chronic lymphocytic leukemia:reports from the 57th American Society of Hematology annual meeting / 白血病·淋巴瘤

With the recent success of the Bruton's tyrosine kinase (BTK) inhibitor, ibrutinib, and the phosphoinositide-3-kinase (PI3K) inhibitor idelalisib in the treatment of patients with relapsed or refractory chronic lymphocytic leukemia (CLL), a number of new agents targeting the B cell receptor (BCR) pathway are in clinical development. In the 57th American Society of Hematology (ASH) annual meeting, great interests are still focused on these two drugs, either monotherapy or combination in the treatment of CLL. On the other hand, SYK inhibitors, new BTK and PI3K antagonists are also coming to the forefront, casting a new light on the treatment of ibrutinib/idelalisb-resistant patients. The progresses of BCR pathway inhibitors in CLL will be summarized in this paper based on the reports in the 57th ASH annual meeting.

Angiopoietin 1 enhances blood-spinal cord barrier permeability via regulating epidermal growth factor receptor pathway substrate 8 expression / 中华神经医学杂志

Objective To explore the role of epidermal growth factor receptor pathway substrate 8 (EpsS) in angiopoietin 1 (Ang1)-induced blood-spinal cord barrier (BSCB) function enhancement in rats.Methods Spinal cord microvascular endothelial cells (SCMECs) were primarily isolated and cultured fiom adult rats to set up the in vitro BSCB models.(1) Transendothelial electrical resistance (TEER) values were measured and Eps8 protein level was detected by Western blotting at different time points (0, 4, 8, 12 h) after Ang1 treatment.Cell lysates untreated with Ang1 were set as controls.(2) The Eps8 expression in SCMECs was silenced by siRNA interference, followed by Ang1 treatment again;the SCMECs were grouped as control siRNA, control siRNA+Ang1, Eps8 siRNA and Eps8 siRNA+Ang1;Western blotting was applied to detect the Eps8 level, TEER values were recorded, endothelial permeability was detected by using sodium fluorescein (Na-F) and Evans-blue albumin (EBA), and then, the F-actin distribution was observed by phalloidine staining.Results (1) TEER values in vitro and Eps8 expression in 4, 8 and 12 h Ang1 treatment groups were both significantly elevated as compared with those in the controls (P＜0.05).(2) The protein level of Eps8 was knocked down by 75％ in Eps8 siRNA group as compared with that in control siRNA group, with significant difference (P＜0.05);while no significant changes of Eps8 expression in the Eps8 siRNA+Ang1 group was noted as compared with that in the Eps siRNA group.Moreover, the Eps8 siRNA group had significantly decreased TEER values, and significantly elevated permeability to Na-F or EBA at different time points as compared with control siRNA group (P＜0.05);differently, the TEER values and permeability to Na-F or EBA of the Eps8 siRNA+Ang1 group did not significantly change as compared with those in the Eps8 siRNA group.F-actin staining also revealed no change between Eps8 siRNA+Ang1 group and Eps8 siRNA group at cell-cell interface, while F-actin arrangement was found to be intensified significantly in the control siRNA+Ang1 group as compared with those in the control siRNA group.Conclusion Ang1 regulates F-actin distribution at rat BSCB by altering the Eps8 expression, which further modulates the barrier function of BSCB.

Relationship between genetic variants in the Toll-like receptor pathway genes and susceptibility of gastric cancer and esophageal squamous cell carcinoma / 中华消化杂志

Objective To investigate the relationship between genetic variants in the Toll-like receptor (TLR) pathway genes and susceptibility of gastric cancer (GC) and esophageal squamous cell carcinoma (ESCC).Methods The data of whole genome association studies of the high-risk population of GC and ESCC in China were analyzed by adaptive rank-truncated product (ARTP) method in pathway and gene level.The associations between single nucleotide polymorphism (SNP) and susceptibility of GC and ESCC were analyzed with additive model of unconditional Logistic regressions.PLINK 1.07 and SPSS 19.0 software were performed for statistical analyses,and ARTP package in R3.0.2 was used for pathway and gene level analysis.Results In gene-level analyses,eight genes were found to be associated with susceptibility of GC (P ＜0.05) and six genes were associated with susceptibility of ESCC (P ＜ 0.05).In single SNP-level analyses,21 SNPs were statistically correlated with susceptibility of GC (P ＜ 0.01),and 11 SNPs were statistically correlated with susceptibility of ESCC (P ＜0.01).Conclusions Some genetic variants in TLR pathway are associated with risk of GC and ESCC.The potential molecular mechanisms need further investigation.

Mechanisms involved in the cytotoxic effects of berberine on human colon cancer HCT-8 cells

Berberine, a constituent of some traditional Chinese medicinal plants, has been reported to have cytotoxicity effects on different human cancer cell lines. There is no available information about the effects and mechanism of action of berberine on human colon cancer cell line HCT-8. In this paper, the cytotoxicity of berberine on HCT-8 cancer cells was investigated by MTT assay, fluorescence microscopy and flow cytometry analysis. Our data revealed that berberine could significantly inhibit the growth of HCT-8 cells in a dose- and time-dependent manner. Morphology of apoptotic cells was studied with acridine orange/ethidium bromide staining. The concentrations of lactate dehydrogenase and both acid and alkaline phosphatases were significantly increased in cell supernatants after berberine treatment, suggesting cell death. Furthermore, flow cytometry analysis showed that berberine could arrest HCT-8 cells at S phase in a time-dependent manner. To further investigate the apoptotic molecular mechanism, reverse transcription-polymerase chain reaction (RT-PCR) and western blotting methods were used. The up-regulated mRNA and/or protein expressions of Fas, FasL, TNF-a, caspase-3 and down-regulation of pro-caspase-3 suggest that the death receptor pathway may be involved in the apoptotic pathway induced by berberine. Decrease of Bcl-2 and increase of Bax in mRNA and/or protein expressions showed that the Bcl-2 family proteins were involved in berberine-induced apoptosis. We also found that berberine-induced apoptosis was associated with an up-regulated expressions of p53 and prohibitin (PHB), and decreased vimentin expression. These results suggest that berberine can suppress cell growth and reduce cell survival by arresting the cell-cycle and by inducing apoptosis of HCT-8 cells.

Gene Expression of Cytokine Pathway,Jak-Stat Signal Pathway and Neuroactive Ligand-receptor Pathway in Patients with Primary Sjgren′s Syndrome / 医学研究杂志

Objective This study is to investigate cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway gene expression pattern of peripheral blood mononuclear cell(PBMC) of primary sjgren syndrome patients.Methods The PBMC sample of 3 patients with sjgren syndrome and 3 healthy volunteers with consistent age were collected.The total RNAs was extracted from the PBMC samples,and reverse transcripted in vitro transcription(IVT),labeled with Cy5/Cy3 and hybridized on the gene chips.After scanning and data extraction with LuxScan 3.0,differentially expressed genes were analyzed with SAM method.The online tool of molecule analysis system(MAS) was used for biological knowledge mining.Results Statistical difference was calculated between the patient and control group in the following three pathways: cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway.Among these,genes of IL-2RA,IL-10 were up-regulated and genes of PF4,GZMA were down-regulated.Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjgren′s syndrome.And the research may provide new target therapy for SS.

Effects of Leukotriene B4-Leukotriene B4 Receptor Pathway in Vascular Immunizing Damage of Kawasaki Disease / 实用儿科临床杂志

Objective To investigate the role of serum in children with Kawasaki disease(KD)in acute stage and ?-globulin role in monocyte cell-produced leukotriene B4(LTB4).Meanwhile,to investigate the effects of the monocyte cell conditioned media(MCM)on the expression of leukotriene B4 receptor 2(BLT2)in endothelial.In order to understand whether LTB4-BLT2 pathway gets involved in vascular damage in KD and the mechanism of ?-globulin in the lessening vascular damage of KD.Methods The concentration of LTB4 in cell culture after the stimulation by serum of healthy children,serum of acute KD and serum of acute KD with ?-globulin were observed,respectively.The expression of BLT2 in the endothelial was determined by flow cytometry.Results 1.The serum of children with KD increased the concentration of LTB4 in MCM(P

The action of death signal receptor pathway of apoptosis in the developmen of gallbladder carcinoma / 中国普通外科杂志

Objective To study the action of death signal receptor pathway of apoptosis in the development of gallbladder carcinoma . Methods Streptavidin biotin peroxidase immunohistochemistry technique was used to study the expression of Fas L in gallbladder carcinoma tissues,and TUNEL method for in situ detection of the number of apoptotic infiltrating lymphocytes around the tumor. Results The positive rates of Fas L in gallbladder carcinoma , gallbladder adenoma, dysplasia of gallbladder epithelium and chronic cholecystis were 84.6%(22/26), 83.3%(15/18) ,100%(3/3) and 55%(11/20), respectively. The positive rate of Fas L in gallbladder carcinoma was significantly higher than in chronic cholecystis (P