Astragalus polysaccharide inhibits IDO1 expression in colon tumor microenvironment to increase intratumoral CD8~+ T cell infiltration / 中国中药杂志

This study aims to investigate the regulatory effects of Astragalus polysaccharide(APS) and APS combined with 5-fluorouracil(5-FU) on indoleamine-2,3-dioxygenase(IDO1) in the colon tumor microenvironment. Sixty Balb/c mice were randomized into a blank group, a model group, an APS group, an APS + 5-FU group, an APS + low-dose 5-FU group, and a 5-FU group. A tumor model was established by subcutaneous transplantation with CT-26 mouse colon cancer cells in other groups except the blank group. After successful modeling, each group was treated with corresponding drugs for 7 days. The general condition, body weight, and tumor volume of the mice were observed and measured daily during the treatment period. The mice were sacrificed at the end of treatment, and the tumor suppression rate and spleen index of the mice were calculated. Western blot and fluorescence quantitative PCR were employed to determine the protein and mRNA levels, respectively, of IDO1 in the tumor tissue of mice. High performance liquid chromatography was employed to measure the levels of tryptophan(Trp) and kynurenine(Kyn) in the tumor tissue of mice. Hematoxylin-eosin(HE) staining was performed to observe the histological changes of the tumor tissue, and immunohistochemistry to detect the changes of CD4 and CD8 expression in the tumor tissue. Compared with that in the model group, the tumor volume of mice in each treatment group significantly reduced. The body weights of mice in APS + 5-FU group and 5-FU group significantly reduced from day 4 to day 7 of treatment. In addition, the APS + 5-FU group and 5-FU group showed significantly decreased spleen index. The protein and mRNA levels of IDO1 were significantly down-regulated in the APS, APS + 5-FU, and APS + low-dose 5-FU groups. The drug interventions significantly increased the Trp content and decreased the Kyn content. The APS + 5-FU group showed significantly reduced infiltration of CD4~+ T lymphocytes and increased infiltration of CD8~+ T lymphocytes. APS inhibited the expression of IDO1 in the colon tumor microenvironment to increase CD8~+ T lymphocyte infiltration, and the combination of APS with 5-FU demonstrated better effect.

Dahuang Lingxian Prescription Regulates Effect of TAK1 and TRAF6 Interaction and Co-localization on Inflammatory Response of Bile Duct Cells / 中国实验方剂学杂志

ObjectiveTo observe the repair effect of Dahuanglingxian prescription （DHLX） on bile duct epithelial cells of rats. To explore whether its mechanism of action is to adjust the mutual binding of transforming growth factor -β （TGF-β） activated kinase 1（TAK1） and tumor necrosis factor receptor-associated factor 6 （TRAF6）， and regulate the activation of the nuclear transcription factor -κB （NF-κB）/mitogen-activated protein kinase （MAPK） signaling pathway. MethodThe 20 SD rats were randomly divided into normal group and DHLX group， 10 rats in each group， were given saline and DHLX （320 mg·kg-1·d-1） for 8 days， to prepare normal serum and DHLX serum. Biliary epithelial cells were extracted from normal SD rats and divided into 9 groups： Normal group， model group （20 mg·L-1）， LPS+DHLX group （20 mg·L-1+10% DHLX）， LPS+PDTC group （20 mg·L-1+200 μmol·L-1）， LPS+SB203580 group （20 mg·L-1+0.5 μmol·L-1）， LPS+PDTC+SB203580 group （20 mg·L-1+200 μmol·L-1+0.5 μmol·L-1）， LPS+PDTC+DHLX group （20 mg·L-1+200 μmol·L-1+10% DHLX serum）， LPS+SB203580+DHLX group （20 mg·L-1+0.5 μmol·L-1+10% DHLX serum）， LPS+PDTC+SB203580 +DHLX group （20 mg·L-1+200 μmol·L-1+0.5 μmol·L-1+10% DHLX serum）. The microscopic observation of morphological changes in each group of cells after drug intervention. Enzyme-linked immunosorbent assay（ELISA） was used to detect the expression of （IL）-1β and IL-6 in each group of cells. Western blot detected the expression levels of TAK1 and TRAF6 proteins in each group of cells， Co-IP detected the interaction between TAK1 and TRAF6， and further observed the distribution and co-localization of TAK1 and TRAF6 using Laser confocal microscope. ResultAfter the action of LPS， the cell synapses are reduced， the cell body becomes significantly rounded and smaller， but the cell morphology of each group tends to be normal after medication. Compared with normal group， the expression levels of IL-1β and IL-6 in model group were significantly increased （P<0.05）， while the expression level of TAK1 was decreased while the expression level of TRAF6 was increased （P<0.05）. The content of TAK1-TRAF6 protein complex showed a decreasing trend， and the two proteins co-located in the cytoplasm. Compared with model group， the expression levels of IL-1β and IL-6 in LPS+DHLX group were significantly decreased （P<0.05）， the expression level of TAK1 was increased and the expression level of TRAF6 was decreased （P<0.05）， the content of TAK1-TRAF6 protein complex was significantly increased （P<0.01）， and the two proteins were significantly co-located in cytoplasm. Compared with LPS+DHLX group， the expression levels of IL-1β and IL-6 in other groups were significantly decreased （P<0.05，P<0.01）. TAK1-TRAF6 protein complex content in each group was significantly decreased after pathway blocker intervention （P<0.05）， while TAK1-TRAF6 protein complex content in each group was significantly increased after pathway blocker combined with DHLX intervention （P<0.05）. Co-localization of the TAK1-TRAF6 in cytoplasm was not obvious. ConclusionIn the LPS-induced inflammatory response of bile duct cells， the binding of TAK1 and TRAF6 showed a weakening trend， but DHLX could reverse the phenomenon， we think the mechanism of action may be related to promoting the mutual binding of TAK1 and TARF6 to inhibit the activation of the NF-κB/MAPK signaling pathway.

Comparison of tranexamic acid pharmacokinetics after intra-articular and intravenous administration in rabbits

Tranexamic Acid [TXA] is commonly administered in total knee arthroplasty for reducing blood loss. There has been a growing interest in the topical use of TXA except intravenous use for prevention of bleeding in TKA. The aim of this study was to develop and validate a HPLC-MS method to detect TXA and apply to compare the pharmacokinetic profile of TXA after intravenous [IV] and topical intra-articular [IA] application of TXA at a dose of 20 mg/kg in rabbits. In order to prove intra-articular administration is better than that of intravenous administration from the point of rabbit pharmacokinetic. Two groups of rabbits [n=6/group] respectively received TXA intra-articularly or intravenously. Blood samples were collected at scheduled time. The concentration of TXA in plasma was determined by a validated HPLC-MS method. Excellent linearity was found between 0.015 and 70.0microg/ml with a lower limit of quantitation [LLOQ]of 0.015microg/ml [r>0.99]; moreover, all the validation data including accuracy and precision [intra- and inter-day] were all within the required limits. The pharmacokinetic parameters in IA and IV group were: C[max]: 30.65+/-3.31 VS 54.05+/- 6.21 fig/ml [p<0.0l]; t[1/2]: 1.26+/-0.05 VS 0.68+/-0.13h [p<0.05]; AUC[0-t]: 42.98+/-7.73 VS 23.39+/-4.14microg/ml- h [p<0.0l], time above the minimum effective concentration [%T > MEC]: 1.5-2.2 VS 0.7-1.2h [p<0.05]. HPLC-MS method is suitable for TXA pharmacokinetic studies. The results demonstrated that topical intra-articular application of TXA showed a reduced peak plasma concentration and prolonged therapeutic drug level compared with intravenous TXA from the point of rabbit pharmacokinetic

Dynamic changes of the epididymal size after vasectomy: An ultrasonographic observation / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the dynamic changes of the epididymal size 1 year after vasectomy.</p><p><b>METHODS</b>Fifty male volunteers received vasoligation. Before and at 1, 2, 3, 6, and 12 months after operation, we measured the size and detected the internal echoes of the epididymis using color Doppler ultrasonography.</p><p><b>RESULTS</b>The bilateral epididymides were both thickened post-operatively in all the 50 cases, with statistically significant differences between the baseline and the 1st month, the 1st and the 2nd month, the 2nd and the 3rd month, or the 3rd and the 6th month after surgery (all P < 0.01), but not between the 6th and the 12th month (P > 0.05).</p><p><b>CONCLUSION</b>Within 6 months after vasectomy, the bilateral epididymides manifested a progressive thickening, but basically restored their balance of secretion-absorption after 6 months.</p>

Triterpenoids and steroids from Excoecaria agallocha / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Excoecaria agallocha L.</p><p><b>METHOD</b>The constituents were isolated and purified by repeated column chromatography and their structures were elucidated by spectroscopic analysis.</p><p><b>RESULT</b>Six triterpenoids including taraxerone (1), beta-amyrin acetate (2), 3beta-[(2E,4E)-6-oxo-decadienoyloxy]-olean-12-ene (3), taraxerol (4), acetylaleuritolic acid (5), and cycloart-22-ene-3beta, 25-diol (6), and three steroids including beta-sitostenone (7), (24R)-24-ethylcholesta-4,22-dien-3-one (8), and beta-sitosterol (9) were isolated and identified from the stems and twigs of the mangrove plant E. agallocha.</p><p><b>CONCLUSION</b>Compounds 5-8 were isolated from E. agallocha for the first time.</p>

Chemical constituents from herb of Gueldenstaedtia stenophylla / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Gueldenstaedtia stenophylla.</p><p><b>METHOD</b>Various chromatographic techniques were used to separate and purify the constituents and structure determination was mainly based on the analysis of the spectroscopic data.</p><p><b>RESULT</b>Seven compounds, including 2', 4, 7-trihydroxy-4'-methoxyisoflavans (1), genkwanin (2), quercetin (3), rutin(4), 12-oleanen-3beta, 22beta, 24-triol (5), betulinic acid (6), and 3, 4-dihydroxybenzoic acid (7) were isolated and identified.</p><p><b>CONCLUSION</b>All these compounds were isolated from the genus Gueldenstaedtia for the first time.</p>