c cancer SGC7901 cells in vitro. However, in vitro studies of the anticancer effect of PAB are rare, and neither the in vivo efficacy of this novel herbal compound against tumors nor its precise molecular mechanism against MDR has been fully investigated. The aim of the present study is to evaluate the anti-neoplastic effect of PAB in vivo, including the reversal of MDR, using a 1 Inhibitory Effect of Pseudolaric Acid B buy Debio1347 injected with 2.56106/ml SGC7901 cells in 0.2 ml of RPMI-1640 medium into the left axillae, and the other 25 mice were injected with SGC7901/ADR cells into the right axillary region under germ-free conditions. Experimental groups and medication Seven days after cell implantation, the tumors became palpable, and then, the two groups injected with two different types of cells were randomly divided into five subgroups: normal saline control group, TWEEN control group, ADR group, PAB group, and PAB+ADR group. For the PAB groups, PAB dissolved in an aqueous solution of 6% polyethylene glycol, 3% ethanol, and 1% Tween80 was administered intraperitoneally daily for 20 days. An identical volume of aqueous solution or NS was injected in the PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19673813 TWEEN control group or NS control group, respectively. For PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19674121 the ADR group, ADR diluted in NS was administered i.p. daily for 20 days. Tumor-bearing mice were administered both PAB and ADR i.p. for the same period in the PAB+ADR group. After treatment, mice from each group were sacrificed, and tumor samples of bilateral axillary regions were weighed and resected for immunohistochemical and western 
blot analyses. This study was performed strictly in accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Committee on the Ethics of Animal Experiments of the Shengjing Hospital Affiliated to China Medical University. Mice were sacrificed under 10% chloral hydrate anesthesia, and all efforts were made to minimize suffering. xenograft model in nude mice and explore whether PAB’s underlying molecular mechanism is related to the inhibition of MDR via the Cox-2/PKC-a/P-gp pathway. Materials and Methods Materials RPMI-1640 culture medium and fetal bovine serum for cell culture were purchased from Gibco BRL, Gaithersburg, MD, USA. Rabbit anti-Cox-2 monoclonal antibody, mouse anti-PKCa and mouse anti-P-gp monoclonal antibodies were purchased from Santa Cruz Corporation, CA, USA. Secondary antibodies against the rabbit and mouse antibodies, streptavidin-peroxidase kits, and 3,3′-diaminobenzidine were obtained from Beijing Zhongshan Biotechnology, China. BCA protein assay kits and SDS-PAGE gel preparation kits were obtained from the Beyotime Institute of Biotechnology, Shanghai, China. PAB was purchased from the Liaoning Institute for Drug Control, China No. 201003. Adriamycin was obtained from Zhejiang Hisun Pharmaceutical Co., Ltd., China No. 120906. Tween80 and polyethylene glycol were purchased from Sigma Chemical, St. Louis, MO, USA. Measurement and calculation of tumor volume and weight Body weight was monitored every day, and two perpendicular diameters of tumors were measured every two days with calipers throughout the treatment period. Tumor volumes were estimated using two-dimensional measurements and calculated as follows: tumor volume = length 6width2 /2. The relative tumor volume was calculated as follows: RTV = /. The antitumor effects of PAB and/or ADR were estimated with the mean inhibiti