Effect of Gegen Qinliantang on SIRT1/FoxO1 Autophagy Pathway in db/db Diabetic Mice / 中国实验方剂学杂志

ObjectiveTo validate the alleviating effect of Gegen Qinliantang （GGQLT） on insulin resistance in db/db diabetic mice by regulating the silent information regulator 1 （SIRT1）/forkhead transcription factor O1 （FoxO1） autophagy pathway. MethodSeventy-five SPF-grade spontaneous type 2 diabetic db/db mice and 15 control db/m mice were selected and maintained on regular feed for one week before measuring blood glucose. They were randomly divided into six groups， with 15 mice in each group. The groups included a normal group （physiological saline， 0.2 g·kg-1）， a metformin group （0.2 g·kg-1）， high-， medium-， and low-dose GGQLT groups （31.9， 19.1， 6.9 g·kg-1）， and a model group （physiological saline， 0.2 g·kg-1）. They were orally treated with corresponding drugs for eight weeks， once daily. Fasting blood glucose （FBG） was measured using a Roche glucometer. Serum levels of high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， triglyceride （TG）， and total cholesterol （TC） were measured using an automated biochemical analyzer. Fasting serum insulin （INS） levels were determined using enzyme-linked immunosorbent assay （ELISA）， and the homeostasis model assessment of insulin resistance （HOMA-IR） was calculated. Western blot was used to detect the expression of Beclin-1， microtubule-associated protein 1 light chain 3 （LC3）， and SIRT1/FoxO1 autophagy pathway-related proteins in liver tissues. Immunohistochemistry was performed to assess the expression of SIRT1， FoxO1， Beclin-1， and LC3B proteins in liver tissues. Transmission electron microscopy was used to observe the formation of autophagosomes in the liver. ResultCompared with the normal group， the model group showed significant increases in FBG， FINS， HOMA-IR， TC， TG， LDL-C， and HDL-C levels （P<0.01）， and significant increases in the expression of SIRT1， Beclin-1， LC3， and FoxO1 proteins in liver tissues （P<0.01）. Transmission electron microscopy revealed the highest number of autophagosomes in the model group. Compared with the model group， the metformin group and the low-， medium-， and high-dose GGQLT groups showed significant decreases in serum FBG， FINS， HOMA-IR， TC， TG， LDL-C， and HDL-C levels （P<0.05， P<0.01）， significant decreases in the expression of SIRT1， Beclin-1， LC3 （P<0.05， P<0.01）， and up-regulated FoxO1 protein （P<0.01）. Transmission electron microscopy showed a reduction in the degree of autophagy in the treatment groups. Compared with the metformin group， the medium- and high-dose GGQLT groups showed significant decreases in FBG， FINS， and TG levels （P<0.01）， significant decreases in the expression of SIRT1， Beclin-1， and LC3 in liver tissues （P<0.05， P<0.01）， and reduced FoxO1 protein （P<0.01）. The high-dose GGQLT group showed reduced HOMA-IR， TC， LDL-C， and HDL-C levels （P<0.05， P<0.01）. Transmission electron microscopy revealed a significant reduction in autophagosomes in the medium- and high-dose GGQLT groups. ConclusionGGQLT can significantly improve glucose and lipid metabolism disorders， alleviate insulin resistance in db/db mice， and prevent and treat type 2 diabetes by activating the SIRT1/FoxO1 autophagy pathway.

Characterization and evaluation of a novel anti-MUC-1 monoclonal antibody: induction of the idiotypic network in experimental mice / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the anti-idiotypic effect induced by a monoclonal antibody.</p><p><b>METHODS</b>A conventional fusion method was used to obtain hybridoma cells producing monoclonal antibody, which were detected by flow cytometry. ELISA were used to detect the humoral response induced by the antibody in mice. Cytotoxic and proliferation experiments were used to detect the cellular response induced by the antibody in mice.</p><p><b>RESULTS</b>CS20 is a MUC-1 specific monoclonal antibody that strongly reacts with MUC-1 antigen expressed on the cell surface of breast cancer cells. The antibody could not kill tumor cells directly through complement-dependent cytotoxicity or antibody-dependent cell-mediated cytotoxicity. However, after 6 administrations of mAb CS20-KLH (keyhole limpet hemocyanin) conjugated to BALB/c mice (n = 5) at a dose of 50 micro g/mouse, anti-idiotypic antibodies and anti-anti-idiotypic antibodies were induced. T cells derived from CS20-KLH-immunized mice responded to mAb CS20, indicating the existence of idiotype-specific T cells.</p><p><b>CONCLUSION</b>These data indicated the possibility of using MUC-1 specific antibody for active immunotherapy of breast cancer.</p>

in vitro Model for Targeting Tumor Vasculature Using Immunoliposome / 中国肿瘤生物治疗杂志

Objective: Based on tumor angiogenesis theory, a target system of tumor endothelial cells was established to investigate the cytotoxicity against tumor vascular endothelium by an drug loaded immunoliposome.Methods: Sterically stabilized liposomes containing doxorubicin were used for conjugation to a monoclonal antibody (MAb) as a drug-vascular targeting device. The MAb M5/114 directly against MHC Ⅱ, which was expressed by vascular endothelial cells was employed. Binding of cholesteryl ether (CHE) labeled anti-MHC Ⅱ immunoliposomes to endothelial cells (SVEC ) in vitro was detected by a binding assay. Cytotoxicity of drug loaded immunoliposome to endothelial cells was determined by MTT assay.Results: At least 3-fold increase in binding for the targeted liposomes was observed. This data suggest highly selective binding of M5/114-immunoliposomes to MHC Ⅱ expressing endothelial cells. Moreover, the cytotoxicity produced by drug loaded M5/114-immunoliposome was greatly enhanced compared with that by non-targeted liposomes. Conclusion: This is the first report using immunoliposome to target endothelial cells. The results demonstrate that immunoliposome directly against tumor vascular endothelial cells might provide a general treatment for solid tumors.