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AIM To analyze and compare HPLC fingerprints of wanai and Artemisiae argyi Levi.et Vant from thirty-one growing areas by multistatistical.METHODS The analysis of 80％ methanol extract of A.argyi was developed on a 30 ℃ thermostatic Agilent TC-C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.2％ methanoic acid) flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 330 nm.RESULTS There were eighteen,twenty-five common peaks in the fingerprints of thirty-one batches of A.argyi,fifteen batches of wanai,respectively,with the similarities all more than 0.900.The similarities of thirty-one batches of samples from different growing area were good and together as a category except samples from Dengzhou city,Luohe city and Anhui province.Fifteen batches of wanai samples got together with Qiai among them.The cumulative contribution rate of the four principal components from A.argyi was 86.049％.Twelve batches of wanai samples had higher scores than Qiai.CONCLUSION This stable and reliable method can be used for the quality control of A.argyi.
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This study was designed to explore the effect of apigenin (Api) on dendritic cell (DCs) maturation and function in murine spleen cells. The single spleen cell was isolated, and then cultured with lipopolysaccharide (LPS) in the present and absence of apigenin. After 24 h, the toxicity of Api and the T cell proliferation were determined by CCK8 kit. In addition, we collected the cell-free supernatants to measure cytokine production using ELISA, collected the cells to determine the DC maturation using flow cytometry. Finally, we purified Api and/or LPS-treated CD11c+ DCs which were pulsed with ovalbumin (OVA)323-339 and then were adoptive transferred into C57BL/6 mice to detect the OVA323-339-specific T cell proliferation and T helper (Th1) and Th2 cell secreting IFN-γ and IL-4 production, respectively. We found that Api did not affect splenocyte viability, but inhibited the production of pro-inflammatory cytokine IL-1β, IL-6 and TNF-α, not anti-inflammatory cytokine IL-10. In addition, Api inhibited the expression of co-stimulatory CD80, CD86 and MHCII of CD11c+ DCs. Finally, compared to LPS+OVA DCs group, DCs from Api and LPS co-treated splenocytes (Api+LPS+DCs) impaired OVA323-339-specific T cell proliferation and the production of IFN-γ and IL-4 in CD4+ T cells, which had the similar responses with OVA+DCs. These data suggest that Api exhibits anti-inflammatory properties via inhibiting DC activation and function, as a new immune-modulator, which may induce immune-tolerance with a benefit to those with chronic inflammation.
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OBJECTIVE We have recently reported that cysteinyl leukotriene (CysLT) signaling plays an important role in microglial interleukin (IL)-1β secretion and subsequent neurotoxicity. The present study aimed to examine microglial morphological changes and the upstream molecular underlying IL-1βproduction in CysLT receptor agonist leukotriene D4 (LTD4)-treated BV2 microglia in vitro. METHODS Twenty-four hours after murine microglial BV2 cells were stimulated with LTD4 (1-100 nmol·L- 1), the cell proliferation and morphology were observed. The expression level of cysteinyl aspartate-specific protease 1 (CASP1) protein was measured by Western blotin BV2 cells. In addition, BV2 cells were pretreated with or without CysLT1 receptor antagonist montelukast for 1 h and the effects of monte-lukaston LTD4-stimulated microglial activation and CASP1 expression were evaluated. RESULTS The number of BV2 cells had an increasing tendency after 24 h treatment with LTD4, but no significant differences were observed between the control and LTD4-treated cells (P>0.05). Under basal and resting conditions, BV2 microglial cells displayed a ramified morphology. However, LTD4 at 100 nmool · L- 1 drove microglial morphological changes from a ramified towards an amoeboid shape. The expression of CASP1 protein was significantly upregulated in 100 nmool·L-1 LTD4-treated BV2 microglia (P<0.01). Furthermore, pretreatment with CysLT1 receptor antagonist montelukast prevented cell morphological changes and suppressed the increased CASP1 expression in LTD4-treated BV2 cells (P<0.05). CONCLUSION CysLT receptor agonist LTD4 induces morphological changes and CASP1 expressionin BV2 microglia, which can be inhibited by CysLT1 antagonist. These results suggest the involvement of CysLT signaling in microglial morphological changes and CASP1 expression.
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<p><b>BACKGROUND</b>L-proline is a natural, nontoxic cryoprotectant that helps cells and tissues to tolerate freezing in a variety of plants and animals. The use of L-proline in mammalian oocyte cryopreservation is rare. In this study, we explored the cryobiological characteristics of L-proline and evaluated its protective effect in mouse oocyte cryopreservation.</p><p><b>METHODS</b>The freezing property of L-proline was detected by Raman spectroscopy and osmometer. Mature oocytes obtained from 8-week-old B6D2F1 mice were vitrified in a solution consisting various concentration of L-proline with a reduced proportion of dimethyl sulfoxide (DMSO) and ethylene glycol (EG), comparing with the control group (15% DMSO and 15% EG without L-proline). The survival rate, 5-methylcytosine (5-mC) expression, fertilization rate, two-cell rate, and blastocyst rate in vitro were assessed by immunofluorescence and in vitro fertilization. Data were analyzed by Chi-square test.</p><p><b>RESULTS</b>L-proline can penetrate the oocyte membrane within 1 min. The osmotic pressure of 2.00 mol/L L-proline mixture is similar to that of the control group. The survival rate of the postthawed oocyte in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG is significantly higher than that of the control group. There is no difference of 5-mC expression between the L-proline combination groups and control. The fertilization rate, two-cell rate, and blastocyst rate in vitro from oocyte vitrified in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG solution are similar to that of control.</p><p><b>CONCLUSIONS</b>It indicated that an appropriate concentration of L-proline can improve the cryopreservation efficiency of mouse oocytes with low concentrations of DMSO and EG, which may be applicable to human oocyte vitrification.</p>
Subject(s)
Animals , Female , Male , Mice , Cryopreservation , Methods , Cryoprotective Agents , Pharmacology , Fertilization in Vitro , Hydrogen-Ion Concentration , Oocytes , Osmotic Pressure , Proline , Pharmacology , Spectrum Analysis, Raman , VitrificationABSTRACT
<p><b>OBJECTIVE</b>To evaluate the significance of upper mediastinal lymph nodes dissection for thyroid carcinoma patients.</p><p><b>METHODS</b>The clinical data of 79 thyroid carcinoma patients who underwent the upper mediastinal lymph node dissection (between January 1984 and December 1998) were retrospectively analysed. There were 45 male and 34 female with a median age of 47 years (range 10 to 74 years). Follow-up was ended on December 31, 2003.</p><p><b>RESULTS</b>Histopathologically, there were 58 (73.4%) papillary carcinoma, 14 (17.7%) medullary carcinoma, and 7 (8.9%) follicular carcinomas. Four of them had poorly-differentiated carcinoma. Upper mediastinal lymph node dissection was carried out in 62 patients through trans-cervical approach, in 10 through an inverted T-shaped incision, and in 7 through a midline sternotomy. Seventy-six patients had 93 neck lymph node dissection procedures, and 47 patients developed paratracheal lymph node metastasis. The overall 5- and 10-year cumulative survival rate was 64.6% and 63.1%, respectively. Mediastinal lymph node recurrence developed only in 10 patients after initial upper mediastinal lymph node dissection. Nine patients died of upper mediastinal lymph node metastasis. Postoperative complications were observed in 11 patients without perioperative death.</p><p><b>CONCLUSION</b>Upper mediastinal lymph node metastasis is most frequently found in papillary thyroid carcinoma. Surgical dissection of upper mediastinal metastatic lymph nodes through either cervical incision or mediastinotomy is safe and effective with low rate of perioperative complications. It may improve the life quality and survival of thyroid carcinoma patients.</p>