Pharmacokinetics of Achyranthes bidentata on adjuvant arthritis rats by microdialysis and UHPLC-MS/MS / 中国中药杂志

To investigate the " drug-guide" effect of Achyranthes bidentata saponins( ABS) and geniposide( GE) in the treatment on adjuvant arthritis( AA) rats. A UHPLC-MS/MS method for the quantitative determination of GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa in rat blood and joint dialysate was established. After single or combined administration with ABS and GE was given to AA rat model,a microdialysis sampling method for rat joint cavity and jugular vein blood vessels was established to collect microdialysis samples. Waters Acquity HSS C_(18) column was used to separate the above four components,with mobile phase as acetonitrile-0. 1% formic acid water as mobile phase for gradient elution. ESI source was adopted for mass spectra in a negative ion scanning mode. Multiple reaction monitoring( MRM) mode was applied to detect the above four components. The methodological results showed that GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa demonstrated a good linear relationship within the concentration ranges of 2-4 000,16-4 096,14-3 584,23-5 888 μg·L-1 respectively. The precision,accuracy,stability and matrix effect of these four ingredients reached the requirements of quantitative analysis of biological samples. The pharmacokinetic results demonstrated that the combined administration of ABS and GE( 60 mg·kg~(-1)+60 mg·kg~(-1)) can increase the degree of GE in joint cavity distribution,and the AUCjoint/AUCplasmwere twice of that of single administration of GE( 60 mg·kg~(-1)),which indicated that ABS might played a vital role in GE's distribution to joint cavity. Moreover,there was no significant difference between the distribution trend of total three ABS and GE in rats. The pharmacodynamics results showed that the combined administration of ABS and GE has stronger effects on paw swelling,arthritis index and synovial pathomorphology of AA rats than single administration of GE,which suggested that ABS might improve GE's anti-inflammatory effect in AA rats. Based on the above results,ABS has a targeting effect in increasing GE's concentration in joint cavity,with a synergy in efficacy.

Silencing effect of cell-specific RNA interference plasmid pPSMAe/p-shNS-ploy(A) loaded by transgenic vector Tf-PEG-PEI targeting nucleostemin on prostate cancer cells in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the transgenic efficiency of non-viral vector Tf-PEG-PEI and the cell specific silencing effect of plasmid pPSMAe/p-shNS-ploy(A) on prostate cancer cells.</p><p><b>METHODS</b>Polyethyleneimine (PEI) was modified by using polyethylene glycol and transferrin to synthesize the non-viral vector Tf-PEG-PEI. NS-specific plasmids pPSMAe/p-shNS-ploy(A) and Tf-PEG-PEI were used to transfect prostate cancer LNCap and PC-3 cells. The changes of cell morphology, proliferation ability and cell cycle were studied after down-regulating the NS gene level.</p><p><b>RESULTS</b>Tf-PEG-PEI was successfully modified. After transfection, the PSMA-expressing LNCaP cells became larger and showed more pseudopodia, having a tendency to differentiate. Their cell proliferation ability was reduced, and the detection of cell cycle showed a decrease of S phase and an increase of G(1) phase after knocking down NS gene. These targets were not changed in non-PSMA-expresing PC-3 cells.</p><p><b>CONCLUSIONS</b>The non-viral vector Tf-PEG-PEI has a high ability to transfer targeted gene into target cells. The cellular specificity of short-hairpin RNA transcription driven by PSMAe/p is confirmed by silencing NS gene. The use of cell specific promoter may be an effective strategy of gene therapy for prostate cancer.</p>

Effect of autophagy-related gene Beclin 1 on growth of xenografts of human lung adenocarcinoma A549 cells in nude mice / 肿瘤

Objective: To investigate the effect of autophagy-related gene Beclin 1 on growth of xenografts of human lung adenocarcinoma A549 cells in BALB/c nude mice. Methods: The recombined plasmids pRNAT-U6.2/Lenti-si423 and pLenex-Beclin 1 were transiently transfected by lipofectin regeant into A549 cells, respectively. The expression levels of Beclin 1 mRNA and protein in A549 cells were detected by real-time fluorogenic quantitative-PCR (RFQ-PCR) and Western blotting, respectively. The proliferation rate of A549 cells transfected with recombinant vector pRNAT-U6.2/Lenti-si423 or pLenex-Beclin 1 was determined by MTT assay. The A549 cells expressing Beclin 1 or with Beclin 1 silencing were subcutaneously injected into right axillary region of nude mice. The growth rate and size of xenograft tumor were observed, and the expression levels of Beclin 1 mRNA and protein in xenograft tumor were determined by RFQ-PCR and immunohistochemistry, respectively. Results: The expression levels of Beclin 1 mRNA and protein were increased in A549 cells transfected with recombinant vector pLenex-Beclin 1, and the cell proliferation in vitro was inhibited. The expression levels of Beclin 1 mRNA and protein were decreased in A549 cells transfected with recombinant vector pRNAT-U6.2/Lenti-si423. As compared with Beclin 1-silencing group, the growth rate of subcutaneous xenograft was slowing down with lower weight and smaller volume and the relative expression levels of Beclin 1 mRNA and protein were both higher in Beclin 1-overexpression group. Conclusion: Autophagy-related gene Beclin 1 can inhibit the growth of xenograft of human lung adenocarcinoma A549 cells in nude mice, and it may become a new target for tumor therapy. Copyright© 2011 by TUMOR.

Determination of beta-endorphin in hypothalamus of rat with primary trigeminal neuralgia / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To determine the level and the role of beta-endorphin in hypothalamus of rat with trigeminal neuralgia.</p><p><b>METHODS</b>The animal model of primary trigeminal neuralgia in rat was established, the contents of beta-endorphin were measured by radioimmunoassay techniques.</p><p><b>RESULTS</b>The beta-endorphin of hypothalamus in experimental group was significantly lower than other groups (P < 0.01).</p><p><b>CONCLUSIONS</b>Beta-endorphin may play important roles in primary trigeminal neuralgia.</p>