New eudesmane sesquiterpenoids from Atractylodis Macrocephalae Rhizoma and their inhibitory activities against SREBPs / 中国中药杂志

Three sesquiterpenoids were isolated and purified from the 95% ethanol extract of Atractylodis Macrocephalae Rhizoma by column chromatography on silica gel, Sephadex LH-20, ODS, and high-performance liquid chromatography(HPLC). Their chemical structures were identified on the basis of spectroscopic analysis and physiochemical properties as(7Z)-8β,13-diacetoxy-eudesma-4(15),7(11)-diene(1), 7-oxo-7,8-secoeudesma-4(15),11-dien-8-oic acid(2), and guai-10(14)-en-11-ol(3). Compounds 1 and 2 are new compounds and compound 3 was obtained from Compositae family for the first time. Compounds 1, 2, and 3 showed weak inhibitory activities against sterol regulatory element-binding proteins(SREBPs).

Knockdown of IOP1 Induces Premature Senescence in Primary HUVECs / 中国生物化学与分子生物学报

Iron-only hydrogenase-like protein 1 (IOP1) is a component of the cytosolic iron-sulfur protein assembly (CIA) machinery. IOP1 has been suggested to be a negative regulator of the hypoxia-inducible transcription factor 1(HIF-1). We previously reported that loss of one copy of NAR1 (the yeast homolog of IOP1) in diploid yeast cells leads to increased sensitivity to oxidative stress and decreased replicative lifespan‚ however‚ the underlying mechanism is still unclear. Recently‚ we found that the IOP1 protein was upregulated in late-passaged primary human umbilical vein endothelial cells (HUVECs) compared with that in early-passaged primary HUVECs‚ which indicated a potential association of IOP1 with cellular senescence. The aim of this study was to investigate the potential function of IOP1 in aging in mammalian cells. The primary HUVECs were transfected with IOP1-specific siRNA and subjected to premature senescence assays. We found that IOP1 knockdown leads to premature senescence and decreased cell proliferative ability (P < 0. 01) in primary HUVECs. Further studies revealed that downregulation of IOP1 resulted in upregulated ROS levels (P < 0. 01)‚ enhanced DNA damage (P<0. 05) and decreased mitochondrial respiration (P<0. 01) along with cell cycle arrest at the G

Retrospective analysis of nutritional risk and nutritional treatment options in inflammatory bowel disease / 肠外与肠内营养

Objective:To understand the nutritional risk in patients with IBD,its related factors and nutritional treatment options.Methods:IBD patients treated in Peking University first hospital from January 2006 to December 2015 were studied.Using the Nutritional risk screening 2002 (NRS2002) nutritional risk assessment of the patients were evaluated.According to the body mass index (BMI),patients were divided into normal BMI group (BMI 18.5 ～ 23.9),low BMI group (BMI ＜ 18.5) and high BMI group (BMI ≥ 24).We analyzed the nutritional risk related factors and compared the difference of nutritional therapy options,regarding the UC and CD patients respectively.Results:A total of 388 patients with IBD were enrolled in the study,with UC 306 and CD 82 patients.The total nutritional risk was 49.5％.Although there was no difference in BMI distribution between UC and CD,CD was more likely to have nutritional risk than UC (CD 64.6％,UC 45.4％,(P =0.002).The nutritional risk of low BMI group was 95.7％.There were no differences in age,sex,and family history in IBD patients for the occurrence of nutritional risk.The more frequently recurrence,severe of disease activity,and the wider rang of disease bring the higher nutritional risk for UC patients.But for CD patients,penetrating type,having a history of surgery and severe of disease activity had higher nutritional risk.Adequate caloric nutrition therapy in patients with CD was 77.4％ higher than that of UC 46.8％,(P ＜ 0.001).It was a main principle of our center that UC patients with severe recurrence should not emphasize enteral nutrition and CD patients should first deal with the contraindications before starting enteral nutrition.Conclusions:IBD patients have a high nutritional risk and CD is more obvious than UC,particularly in low BMI patients.The nutritional risk of patients with UC and CD has its own associated factors.It is safe to treat IBD patients with enteral nutrition as long as the indications and contraindications were well controlled.

Induction of robust senescence-associated secretory phenotype in mouse NIH-3T3 cells by mitomycin C / 生理学报

Senescence-associated secretory phenotype (SASP) is often a concomitant result of cell senescence, embodied by the enhanced function of secretion. The SASP factors secreted by senescent cells include cytokines, proteases and chemokines, etc, which can exert great influence on local as well as systemic environment and participate in the process of cell senescence, immunoregulation, angiogenesis, cell proliferation and tumor invasion, etc. Relative to the abundance of SASP models in human cells, the in vitro SASP model derived from mouse cells is scarce at present. Therefore, the study aimed to establish a mouse SASP model to facilitate the research in the field. With this objective, we treated the INK4a-deficient mouse NIH-3T3 cells and the wildtype mouse embryonic fibroblasts (MEF) respectively with mitomycin C (MMC), an anticarcinoma drug which could induce DNA damage. The occurring of cell senescence was evaluated by cell morphology, β-gal staining, integration ratio of EdU and Western blot. Quantitative RT-PCR and ELISA were used to detect the expression and secretion of SASP factors, respectively. The results showed that, 8 days after the treatment of NIH-3T3 cells with MMC (1 μg/mL) for 12 h or 24 h, the cells became enlarged and the ratios of β-gal-positive (blue-stained) cells significantly increased, up to 77.4% and 90.4%, respectively. Meanwhile, the expression of P21 protein increased and the integration ratios of EdU significantly decreased (P < 0.01). Quantitative RT-PCR detection showed that the mRNA levels of several SASP genes, including IL-6, TNF-α, IL-1α and IL-1β increased evidently. ELISA detection further observed an enhanced secretion of IL-6 (P < 0.01). On the contrary, although wildtype MEF could also be induced into senescence by MMC treatment for 12 h or 24 h, embodied by the enlarged cell volume, increased ratios of β-gal-positive cells (up to 71.7% and 80.2%, respectively) and enhanced expression of P21 protein, the secretion of IL-6 displayed no significant change. Our study indicated that, although MMC could induce senescence in both mouse NIH-3T3 cells and wildtype MEF, only senescent NIH-3T3 cells displayed the canonical SASP phenomena. Current study suggested that senescent NIH-3T3 cells might be an appropriate in vitro SASP model of mouse cells.

Rapid separation and identification of multiple constituents in Danhong Injection by ultra-high performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry / 中国天然药物

To characterize and identify multiple constituents in Danhong injection (DHI), a fast ultra-high performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC-ESI-QTOF/MS) method was established and validated in the present study. A total of 63 compounds, including 33 phenolic acids, 2 C-glycosyl quinochalcones, 6 flavonoid O-glycosides, 4 iridoid glycosides, 6 organic acids, 5 amino acids, and 3 nucleosides, were identified or tentatively characterized. In conclusion, the UHPLC-ESI-QTOF/MS method is useful and efficient for in-depth structural elucidation of chemical compounds in complex matrices of herbal medicines such as DHI.

The Involvement of Ca(2+) Signal Pathways in Distal Colonic Myocytes in a Rat Model of Dextran Sulfate Sodium-induced Colitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Disrupted Ca2+ homeostasis contributes to the development of colonic dysmotility in ulcerative colitis (UC), but the underlying mechanisms are unknown. This study aimed to examine the alteration of colonic smooth muscle (SM) Ca2+ signaling and Ca2+ handling proteins in a rat model of dextran sulfate sodium (DSS)-induced UC.</p><p><b>METHODS</b>Male Sprague-Dawley rats were randomly divided into control (n = 18) and DSS (n = 17) groups. Acute colitis was induced by 5% DSS in the drinking water for 7 days. Contractility of colonic SM strips (controls, n = 8 and DSS, n = 7) was measured in an organ bath. Cytosolic resting Ca2+ levels (n = 3 in each group) and Ca2+ transients (n = 3 in each group) were measured in single colonic SM cells. Ca2+ handling protein expression was determined by Western blotting (n = 4 in each group). Differences between control and DSS groups were analyzed by a two-sample independent t-test.</p><p><b>RESULTS</b>Average tension and amplitude of spontaneous contractions of colonic muscle strips were significantly enhanced in DSS-treated rats compared with controls (1.25 ± 0.08 g vs. 0.96 ± 0.05 g, P= 0.007; and 2.67 ± 0.62 g vs. 0.52 ± 0.10 g, P= 0.013). Average tensions of carbachol-evoked contractions were much weaker in the DSS group (1.08 ± 0.10 g vs. 1.80 ± 0.19 g, P= 0.006). Spontaneous Ca2+ transients were observed in more SM cells from DSS-treated rats (15/30 cells) than from controls (5/36 cells). Peak caffeine-induced intracellular Ca2+ release was lower in SM cells of DSS-treated rats than controls (0.413 ± 0.046 vs. 0.548 ± 0.041, P= 0.033). Finally, several Ca2+ handling proteins in colonic SM were altered by DSS treatment, including sarcoplasmic reticulum calcium-transporting ATPase 2a downregulation and phospholamban and inositol 1,4,5-trisphosphate receptor 1 upregulation.</p><p><b>CONCLUSIONS</b>Impaired intracellular Ca2+ signaling of colonic SM, caused by alteration of Ca2+ handing proteins, contribute to colonic dysmotility in DSS-induced UC.</p>

Anti-inflammatory and anti-arthritic effects of Guge Fengtong Formula: in vitro and in vivo studies / 中国天然药物

Rheumatoid arthritis (RA) is the most common inflammatory arthritis and a major cause of disability. Presently, the clinical therapeutic medicines for inflammatory and arthritic diseases are unsatisfactory due to severe adverse effects or ineffectiveness. The Guge Fengtong formula (GGFT), containing the standardized extracts of Dioscoreae Nipponicae Rhizoma, Spatholobi Caulis, and Zingiberis Rhizoma, has long been used for RA treatment by Chinese doctorsin China. However, the detailed anti-inflammatory and anti-arthritic activity of GGFT has not been reported so far. In the present work, we aimed to evaluate the anti-inflammatory and anti-arthritic effects of GGFT using three in vivo animal models, and tried to uncover its preliminarythe underlying mechanism of action mechanism in RAW 264.7 macrophages. The obtained results indicated that GGFT significantly attenuated ear edema, decreased carrageenan-induced paw edema, reduced the arthritis score, and reversed the weight loss of the complete Freund's adjuvant (CFA)CFA-injected rats. Additionally, marked decrease of in synovial inflammatory infiltration and synovial lining hyperplasia in the joints and decline of inflammatory factors (TNF-α and IL-1β) in the serum were observed in the GGFT-treated rats. In lipopolysaccharide-activated RAW264.7 macrophages, GGFT reduced the production of NO, PGE2, and IL-6, and inhibited the expression of iNOS, COX-2, and NF-κB expression. Our results demonstrated that GGFT possessed considerable anti-inflammatory activity and have had potential therapeutic effects on adjuvant induced arthritis in rats, which provided providing experimental evidences for its traditional application in the treatment of RA and other inflammatory diseases.

Diagnostic ion filtering strategy for chemical characterization of Guge Fengtong Tablet with high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry / 中国天然药物

The present study was designed to characterize the chemical constituents of Guge Fengtong Tablet (GGFTT). Based on the chromatographic retention behavior, fragmentation pathways of chemical components and the published literatures, a diagnostic ion filtering strategy with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (HPLC-ESI-Q-TOF/MS) was established to identify the multiple bioactive constituents of GGFTT. The rapid identification of forty-seven components, including 18 phenolic acids, 8 saponins, 14 gingerol-related compounds, and 7 diarylhepatonoids, was accomplished using this newly developed method. The coupling of HPLC-ESI-Q-TOF/MS with the diagnostic ion filtering strategy was useful and efficient for the in-depth structural elucidation of chemical compounds of GGFTT.

Recombinant human thrombopoietin in combination with cyclosporin A as a novel therapy in corticosteroid-resistant primary immune thrombocytopenia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The management of patients with refractory immune thrombocytopenia (ITP) is challenging, as there is no standard treatment option. The aim of this study was to investigate the efficacy of recombinant human thrombopoietin (rhTPO) in combination with cyclosporin A (CsA) for the management of patients with corticosteroid-resistant primary ITP.</p><p><b>METHODS</b>Thirty-six patients with corticosteroid-resistant ITP were randomly divided into an observation group and control group. In the observation group, 19 patients received subcutaneous injection of rhTPO at a dose of 1 µg/kg (300 U/kg) once daily up to day 14. Simultaneously they also received oral CsA at a dose of 1.5-2.0 mg/kg twice daily for three months. In the control group, rhTPO alone was administered subcutaneously at 1 µg/kg once daily in the other 17 ITP patients for 14 consecutive days and then the treatment was withdrawn.</p><p><b>RESULTS</b>There was no significant difference in the response rate at the end of the first week after treatment initiation between the observation group and the control group (63.2% vs. 58.8%, P > 0.05), neither was there at the end of the second week (89.5% vs. 94.1%, P > 0.05). However, the relapse rate in the observation group was significantly lower than that in control group at the end of the first (17.7% vs. 50.0%, P < 0.05), second (29.4% vs. 68.8%, P < 0.05) and the third month (29.4% vs. 87.5%, P < 0.01). In addition, rhTPO plus CsA were well tolerated and adverse events recorded were mild.</p><p><b>CONCLUSIONS</b>Combination therapy with rhTPO and CsA was effective in the management of patients with corticosteroidresistant ITP, with a relatively short time to response and low recurrence rate. It might be considered as a potential secondline treatment regimen for ITP.</p>

Effect of the 5-HT4 receptor and serotonin transporter on visceral hypersensitivity in rats

Visceral hypersensitivity plays an important role in motor and sensory abnormalities associated with irritable bowel syndrome, but the underlying mechanisms are not fully understood. The present study was designed to evaluate the expression of the 5-HT4 receptor and the serotonin transporter (SERT) as well as their roles in chronic visceral hypersensitivity using a rat model. Neonatal male Sprague-Dawley rats received intracolonic injections of 0.5% acetic acid (0.3-0.5 mL at different times) between postnatal days 8 and 21 to establish an animal model of visceral hypersensitivity. On day 43, the threshold intensity for a visually identifiable contraction of the abdominal wall and body arching were recorded during rectal distention. Histological evaluation and the myeloperoxidase activity assay were performed to determine the severity of inflammation. The 5-HT4 receptor and SERT expression of the ascending colon were monitored using immunohistochemistry and Western blot analyses; the plasma 5-HT levels were measured using an ELISA method. As expected, transient colonic irritation at the neonatal stage led to visceral hypersensitivity, but no mucosal inflammation was later detected during adulthood. Using this model, we found reduced SERT expression (0.298 ± 0.038 vs 0.634 ± 0.200, P < 0.05) and increased 5-HT4 receptor expression (0.308 ± 0.017 vs 0.298 ± 0.021, P < 0.05). Treatment with fluoxetine (10 mg·kg-1·day-1, days 36-42), tegaserod (1 mg·kg-1·day-1, day 43), or the combination of both, reduced visceral hypersensitivity and plasma 5-HT levels. Fluoxetine treatment increased 5-HT4 receptor expression (0.322 ± 0.020 vs 0.308 ± 0.017, P < 0.01) but not SERT expression (0.219 ± 0.039 vs 0.298 ± 0.038, P = 0.654). These results indicate that both the 5-HT4 receptor and SERT play a role in the pathogenesis of visceral hypersensitivity, and its mechanism may be involved in the local 5-HT level.

Determination of platelet-associate autoantibodies against platelet-specific receptors by cytometric bead array and its clinical application in idiopathic thrombocytopenic purpura / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish a method for detecting platelet-associate autoantibodies against platelet-specific receptors using cytometric bead array, and compare the clinical usefulness of this method and modified indirect monoclonal antibody immobilization of platelet antigen technique (MAIPA) in the differential diagnosis of idiopathic thrombocytopenic purpura (ITP) from non-immune thrombocytopenic purpura (non-ITP).</p><p><b>METHODS</b>The microbeads were coated with monoclonal antibodies against glycoprotein IIb/IIIa (CD41a), platelets were isolated from blood samples, then platelet lysate was incubated with the coated microbeads, and R-Phycoerythrin-conjugated goat-antihuman IgG polyclonal antibodies, finally analyzed with flow cytometry. GP IIb/IIIa autoantibodies in sample plasma were measured by modified indirect MAIPA at the same time.</p><p><b>RESULTS</b>The individual fluorescence level was calculated as the ratio to the three controls. The mean ratios were 3.36 (range 0.84 - 22.94) in the ITP group, 1.16 (range 0.67 - 5.59) in the non-ITP patient and 1.08 (range 0.72 - 1.76) in the healthy controls. There was a highly significant difference (P <0.01) between the ITP patients and either the non-ITP patients or the normal controls. If the up limit of healthy controls was set as cutoff value, ratio of greater than 1.76 was considered positive. Cytometric bead array had a sensitivity of 71.43%, a specificity of 94.28% and a positive predictive value of 95.24% for the diagnosis of ITP, the sensitivity being higher than that of modified indirect MAIPA' s (5179%) (chi2 = 4.57, P <0.05). The ROC curve showed the discriminative validity of cytometric bead array was 0.916.</p><p><b>CONCLUSION</b>Flow cytometric bead method for detection of platelet-associate autoantibodies against platelet-specific receptors is a more rapid, better reproducibility and higher sensitivity than modified MAIPA, and has a potential value in promoting the diagnosis of ITP and guiding treatment.</p>

Inhibitory effect and kinetic analysis of sodium quercetin-7,4'-disulphate on recombinant human protein kinase CK2 holoenzyme / 药学学报

<p><b>AIM</b>To study the direct effect and kinetics of sodium quercetin-7,4'-disulphate (SQDS) on recombinant human protein kinase CK2 holoenzyme.</p><p><b>METHODS</b>The recombinant human CK2 holoenzyme activity was assayed by detecting incorporation of 32P of [gamma-32P] ATP into the substrate in various conditions.</p><p><b>RESULTS</b>The recombinant human CK2 was a second messenger (Ca2+, cAMP and cGMP) independent protein kinase. The characterization and function of the reconstituted holoenzyme were consistent with those of native CK2. SQDS was shown to strongly inhibit the holoenzyme activity of recombinant human protein kinase CK2 with an IC50 of 4.4 mumol.L-1, which was more effective than DRB and A3, known CK2 special inhibitors. Kinetic studies of SQDS on recombinant human CK2 showed: the inhibition was competitive with ATP and noncompetitive with casein.</p><p><b>CONCLUSION</b>SQDS is a potent inhibitor of protein kinase CK2. This study provide experimental basis for the development of more effective inhibitors of CK2 and for clinical application of SQDS in the future.</p>