OBJECTIVE(99)Tc(m) labeled C2A domain of synaptotagmin I ((99)Tc(m)-Syt I-C2A) is used for noninvasive detection of vulnerable atherosclerotic plaque.

METHODSRecombinant C2A domain of synaptotagmin I, overexpressed in E. Coli, was thiolated with 2-iminothiolane (2-IT) and labeled with (99)Tc(m). Atherosclerotic plaques were produced in 5 rabbits by deendothelialization of the abdominal aorta and the rabbits were fed with cholesterol diet for 3 months. Three rabbits not manipulated served as normal controls. All animals were injected with (99)Tc(m)-Syt I-C2A and underwent in vivo imaging thereafter. Aortas were then explanted for ex vivo imaging and histological characterization.

RESULTSIn deendothelialized animals, intense radio-uptake in abdominal aorta, showed by gamma camera at 2 h after injection, was visualized and T/B was 3.25 +/- 0.51 by ROI measurement, quantitative uptake ratio of abdominal aortas with atherosclerotic lesions to thoracic aortas was 8.39 +/- 1.74 in ex vivo imaging. The mean uptake in specimens of abdominal aortas with lesions was 12.6-fold higher than in control abdominal aortas, and 10.2-fold higher than in thoracic aortas of deendothelialized animals by gamma-counter.

CONCLUSION(99)Tc(m)-Syt I-C2A has a high affinity for vulnerable atherosclerotic plaque and is a suitable a gent for the noninvasive detection of vulnerable atherosclerotic plaque.

Animals ; Atherosclerosis ; diagnostic imaging ; pathology ; Disease Models, Animal ; Immunoglobulin Fab Fragments ; Isotope Labeling ; Male ; Rabbits ; Radionuclide Imaging ; Synaptotagmin I ; immunology ; Technetium

OBJECTIVE: To explore the effects of Baisong tablets (BST) on synapse protein synatotagmin (SYT) and synaptophysin (SYN) of hippocampus in chronic stress depression in rats. METHODS: Twenty eight male Sprague-Dawley rats were randomly allocated to 4 groups: a normal control group,a model group,a fluoxetine (FXT) group and a BST group. The normal control rats were fed in a natural environment. Rats of the model, FXT and BST groups were singly housed and given an chronic unpredicted sequence of mild stressors. The distribution and expression differences of SYT and SYN in the hippocampus of rats in different groups were investigated with in situ hybridization and immunoblotting. RESULTS: Expressions of SYT and SYN in the hippocampus of model rats were significantly reduced, compared with that of the normal control (P<0.05); and the expressions of SYT and SYN were significantly increased in the hippocampus of the FXT and BST groups, compared with that of the model group (P<0.05). CONCLUSION The expressions of SYT and SYN protein and their mRNA decrease in the hippocampus of stress-model rats. BST can up-regulate their expression.
Animals ; Antidepressive Agents ; therapeutic use ; Depression ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Hippocampus ; metabolism ; Membrane Glycoproteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological ; Synaptotagmin I ; biosynthesis ; genetics

UNLABELLEDObjective To evaluate the efficacy of 99mTc-labeled C2A probe in detection of apoptosis of non-small cell lung cancer (NSCLC) cells after chemotherapy.

METHODSImaging studies were performed in NSCLC H460-bearing mice. The mice were divided into 2 groups: the paclitaxel-treated group and control group. 99mTc-C2A was injected intravenously at 12, 24, 48 and 72 h after chemotherapy. Images were acquired at 3 h and 6 h after injection using a pinhole collimator. The regions of interest (ROI) were drawn in tumor area and contralateral nomal tissue, and the ratio of T/NT were caculated. The tumor sections were stained by HE and TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-nick-end labeling) staining to confirm the presence of apoptosis. Activated caspase-3 was also analyzed with flow cytometry.

RESULTSLittle uptake of 99mTc-C2A was found in baseline images, but tumor uptake increased very much after chemotherapy, the T/NT ratio was 1.79 +/- 0.34, 2.23 +/- 0.33 and 2.78 +/- 0.34, respectively. The T/NT ratio of control was 1.48 +/- 0.23. Tumor uptake (% ID/g) of 99mTc-C2A in chemotherapy groups were 2.82 +/- 0.90, 3.13 +/- 0.48 and 3.52 +/- 1.18, respectively. Tumor uptake (% ID/g) in the control group was 1.21 +/- 0.51. It in paclitaxel-treatment groups were 2.82 +/- 0.90, 3.13 +/- 0.48 and 3.51 +/- 1.18, respectively, significantly higher than that in untreated mice. Furthermore, the uptake of 99mTc-C2A correlated well with apoptotic index (r = 0.56, P < 0.01), and activated caspase-3 (r = 0.59, P < 0.01).

CONCLUSIONOur preliminary results demonstrated that 99mTc-C2A imaging in vivo for detection of cell death in solid tumors is feasible and well correlated with TUNEL staining and activated caspase-3. The C2A holds promise and warrants further development as a molecular probe to early predict cancer treatment efficacy.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; metabolism ; pathology ; Caspase 3 ; metabolism ; Flow Cytometry ; Humans ; In Situ Nick-End Labeling ; Lung Neoplasms ; drug therapy ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Paclitaxel ; pharmacology ; therapeutic use ; Synaptotagmin I ; chemistry ; metabolism ; Technetium ; administration & dosage ; chemistry ; Xenograft Model Antitumor Assays

AIMTo investigate the effect of ginkgolide B on the proliferation of VSMC and the secretion of chemokines by U937 cells stimulated by oxLDL or PAF. In addition, to analyze whether the effect of oxLDL is mediated through PAF receptor.

METHODSUsing 3H-Tdr incorporation assay, the proliferation of VSMC was measured. The protein and mRNA level of MCP-1 and IL-8 in U937 cells were determined by RT-PCR and ELISA. Using Western blotting the p65 and IkappaB was quantified. The binding of oxLDL to U937 cell was measured by a radio-ligand binding assay of 3H-PAF.

RESULTSGinkgolide B inhibited, in dose-dependent manner, the proliferation of VSMC and the secretion of chemokines by U937 cells stimulated by oxLDL, and inhibited the oxLDL-induced p65 activation and depletion of IKappaB. oxLDL inhibited PAF binding to U937 cells.

CONCLUSIONGinkgolide B, as a PAF antagonist, possesses the effect of inhibiting the proliferation of VSMC and the secretion of chemokines by U937 cells stimulated by oxLDL in vitro. The effect of oxLDL is, at least in part, mediated through PAF receptor.

Animals ; Aorta, Thoracic ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chemokine CCL2 ; biosynthesis ; genetics ; Diterpenes ; isolation & purification ; pharmacology ; Dose-Response Relationship, Drug ; Ginkgo biloba ; chemistry ; Ginkgolides ; Humans ; I-kappa B Proteins ; metabolism ; Interleukin-8 ; biosynthesis ; genetics ; Lactones ; isolation & purification ; pharmacology ; Lipoproteins, LDL ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Plants, Medicinal ; chemistry ; Platelet Activating Factor ; antagonists & inhibitors ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Synaptotagmin I ; metabolism ; U937 Cells