To investigate the effect and mechanism of intravascular brachytherapy with 192 Ir on expression of type I collagen after angioplasty. Restenosis model of domestic miniswine was employed.After angioplasty, the iliac arteries were randomized to brachytherapy group ( n =6) , which were treated with 20 25Gy of 192 Ir, and control group ( n =18). The target vessels were harvested at the 12 th week and the 24 th week after angioplasty. Immunohistochemistry and in situ hybridization were used to detect proteins of type I collagen, metalloproteinase 1 (MMP 1) and its tissue inhibitor (TIMP 1), and mRNA expression of type I collagen. The results showed that the protein and mRNA of type I collagen, and the ratios of TIMP 1/MMP 1 were lower in brachytherapy group than in control group. The peak of mRNA expression of type I collagen was at the 24 th week in control group and at the 12 th week in brachytherapy group, respectively. It suggested that intravascular brachytherapy with 192 Ir affects the metabolism of extracellular matrix by inhibiting the synthesis of type I collagen and regulating the activities of MMP 1 and TIMP 1.

Objective To investigate the potential value of VEGF-C targeted ultrasmall superparamagnetic particles of iron oxide (USPIO)molecular probe in specific detection of hepatocellular carcinoma (HCC)in a rat model using MRI.Methods The targeted probe was synthesized by conjugating VEGF-C antibody with amino modified USPIO.Cell counting kit-8 assay was conducted to as-certain the probe’s effect on the growth of HepG2 cells.Rat models with HCC were divided into two groups (targeted group with VEGF-C-USPlO and a contrast with USPIO)with 3 rats for each group at random.Pre-and post-contrast enhanced MR imaging with different time points of 0.5,1 and 1.5h was performed with an injection into caudal vein.The signal intensities of the tumor on T2 WI and T2 * WI were measured,and the differences of the signal intensities between pre-and post-enhancements or between both groups were analyzed.The iron particles within the tumors in two groups were confirmed by Prussian blue iron staining.The ex-pression of VEGF-C in HCC was proved by immunohistochemistry.Results The signal intensities of HCC on T2 WI and T2 * WI af-ter VEGF-C-USPI0 injection were decreased obviously with a minimum value at 1 h ,indicating a significant difference (P <0.05). However,those in USPIO group were decreased less without statistical differences (P >0.05).Statistical differences in signal inten-sity on T2 * WI after enhancement between both groups were also showed (P <0.05).Prussian blue staining results showed more iron particles within the tumor tissues in VEGF-C-USPI0 group,whereas less ones in USPIO group.Immunohistochemical results showed that VEGF-C was over expressed in cytoplasm and membrane.Conclusion VEGF-C-USPI0 molecular probes can initiatively target to the liver cancer in rat models with expressed VEGF-C,which may help to achieve the specific MR imaging of HCC,indica-ting a potential of the metastasis.

BACKGROUND: Bone marrow-derived mesenchymal stem cells (BMSCs) can not only differentiate into multiple nonhematopoietic cell lineages, but seek out damaged tissues and repair them as well. Hence, they were largely studied for their potential clinical use. However, their biological characteristics have not been fully discovered. OBJECTIVE: To compare the biological characteristics of BMSCs of different species cultured in vitro, in order to provide basis for the clinical research of stem cell therapy.DESIGN: Randomized controlled observation was designed.SETTING: Medical Research Department of General Hospital of Guangzhou Military Area.MATERIALS: The experiment was performed in Medical Research Department of General Hospital of Guangzhou Military Area from June 2004 to July 2005. Thirty SD rats weighing (160±20) g, aged 35 to 40 days, 30 Kunming mice weighing (16.0±2.0) g, aged about 40 days, 8 New Zealand white rabbits weighing (2.0±0.2) kg, aged 80 to 90 days and 10 healthy volunteers (25-32 years old) were selected. All the animals were of clean grade, which were purchased from the Animal Center of Southern Medical University.METHODS: The BMSCs of mice and rats were prepared according to the protocol developed in the Caplan laboratory, while those of rabbits and human were isolated from bone marrow suspension obtained by iliac puncture.The morphology of BMSCs was observed by light microscope and transmission electron microscope. Cell growth curve was tested by MTT. Expression of Stro-1 was analyzed by immunofluorescence cytochemistry and flow cytometry. To evaluate the specific response of BMSCs to osteogenic supplements(10 nmol/L dexamethasone, 10 mmol/L β-glycerophosphate,and 50 mg/L ascorbic acid), the activity of alkaline phosphatase (AKP) activity was tested by a commercial kit. Expression of osteocalcin was examined by immunocytochemistry and hydroxyapatite crystals were shown by von Kossa staining. Adipogenic differentiation was evaluated by estimating percent of cells containing Oil Red-O- stained oil droplets.MAIN OUTCOME MEASURES: ① Morphological observation and growth situation of BMSCs. ② Expression of Stro-1: BMSC marker. ③ Differentiation in osteogenic medium.RESULTS: ①The morphology of adherent BMSCs ofthose four species observed by optic microscopy was obviously different. When they became mature or aged, the mouse cells turned into flat shape, irregularly polygonal, fell to pieces and deposited on the flask-bottom, while the rat, rabbit and human cells would enlarge and become polygonal, vacuoles would appear in their cytoplasm, finally, the cells were detached from the flask-bottom, floating off like cotton wool. The cultures of different species also had some commonness, such as poly-layer growing manner, without contact inhibition and consisting of two groups. Cells of one group grew into colonies from single cells and expanded quickly, while cells of the other group were sporadic and did not proliferate. Electron microscopy revealed that all of the primary cells had microvilli and that they could be divided into two subpopulations according to their ultrastructures. Some cells were rich in organelles and most chromatin was euchromatin, while the other subpopulation cells had much fewer organelles and more heterochromatin. Growth curves of BMSCs of different species were almost the same. ② The positive rate of human adherent bone marrow-derived cells for Stro-1: BMSC marker was (91.4±8.3) %, and that of mouse adherent cells was (83.5±6.2) % .③Treated with osteogenic supplements, mouse BMSCs differentiated into adipose tissue, rabbit ones died, while rat and human ones differentiated into osteocytes. BMSCs also demonstrated spontaneous differentiation in vitro.CONCLUSION: Mouse, rat, rabbit and human BMSCs can be easily expanded in vitro, although the harvest of the current method is a mixture of mesenchymal cells with various maturities, most of which are poor-differen-tiated cells. BMSCs of those species are different in morphology and response to the same inductive supplements. Therefore, in order to establish a kind of stem cell therapy, it is necessary not only for evidence from animal experiments but for that from human experiments as well.

BACKGROUND: There are some studies proposing cell membrane may be one of target cell regions for electromagnetic biological effects. However, reports responsible for cellular membrane permeability and cellular biological effects after electromagnetic irradiation are few. OBJECTIVE: This study was designed to investigate the correlations of low-power millimeter wave irradiation to cellular membrane permeability and apoptosis of HL60.DESIGN: Controlled experiment.SETTING: Thus study was performed at the Medical Experimental Central Laboratory, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA between November 2006 and April 2007. MATERIALS: The human leukemic cell lines HL60 were kindly provided by Medical Experimental Center of Guangzhou Military Area Command of Chinese PLA and HD-413.2HPSG 100 millimeter wave irradiation generator was developed by Xi'an Hengda Microwave Technology Development Company, China. METHODS: HL60 cells were irradiated by millimeter wave at frequency of 41.32 GHz and mean power density of 2 mW/cm2, and divided into five groups according to the irradiation time (0, 15, 30, 45, 60 minutes groups).MAIN OUTCOME MEASURES: Lanthanum tracing was used to observe intracellular and extracellular lanthanum particles distribution and evaluate the change of cellular membrane permeability; ultrastructure and morphological characteristics of HL60 cells were observed through an transmission electron microscope; Quantitative analysis of apoptotic cells was performed by in situ marking method. RESULTS: In the 45 and 60 minutes groups, a small amount of lanthanum particles in the cytoplasm, swollen mitochondria, expanded rough endoplasmic reticulum, and obvious apoptosis were detected by ultrastructure observation. TUNEL staining showed, compared with the 0 minute control group, the apoptosis rate showed a trend of elevation in all the irradiation groups, particularly in the 45 and 60 minutes groups. CONCLUSION: Low-power millimeter wave irradiation on HL60 cells can lead to an increase in cellular membrane permeability of HL60, which may be one of the primary causes for promoting apoptosis and producing other biological effects.

Objective To explore the roles of humoral immunity in severe acute respiratory syndrome (SARS). Methods The pathological changes in the SARS autopsy case and a case of lung puncture were observed by light microscopy and electron microscopy. The autoantibody and immune complexes in the sample tissues were detected by immunohistochemical staining and histochemical staining. Results There were severely diffusive damages in the lungs. The endothelial cells in blood were injured. Fibrinoid necroses of blood vessels were observed in several organs and injury of immune organs and massive autoantibody and immune complexes were found in pathologically changed tissues. Electron dense deposits were observed in the basement membrane of blood vessels and mesangial matrix of glomerulus by electron microscopy. Conclusion The SARS viruses not only cause direct damages to the tissues but also lead to immune complex mediated hypersensitivity which in turn gives rise to a large number of tissue lesions. This indicates that the humoral immunity plays an important role in the process of organ damages.

Aim To elucidate the therapeutic effect of ginsenosides, berberine and jasminoidin after given a-lone or treatment with combination on the focal cerebral ischemia rats and study the compatibility mechanism. Methods We determined 12 endogenous amino acids in serum of rats after cerebral ischemia over 12 hours with RRLC-QQQ to evaluate the integrated role of YQJD at the dosage of 25 mg·kg-1 and 5 mg·kg-1 . Generally accepted methods were used, including be-havior test, One-Way AVONA, PLS-DA, as well as PCA to evaluate the injury induced by focal cerebral is-chemia. Results The score of neurological deficits and the level of five amino acids, namely Glu, Asp, Met, Hcy, Phe in the combination of ginsenosides, berberine and jasminoidin group in the dosage of 25 mg ·kg-1 and 5 mg·kg-1 significantly decreased (P<0. 05, P<0. 01) compared to those of model group. For another, the largest contribution group in the three principal components of PC1 , PC3 , PC4 at the dosage of 25 mg/kg and the six principal components PC1 ~PC5, PC7 in 5 mg·kg-1 was the combination of gin-senosides, berberine, jasminoidin group. Conclusions The results suggest that the efficacy of the combina-tion of ginsenosides, berberine and jasminoidin is su-perior to the combination of two or any single compo-nent, which can significantly improve the metabolic disorder of the endogenous amino acid after cerebral is-chemia. And it could be speculated that ginsenosides may play a more important role than berberine and jas-minoidin in regulating the level of amino acid metabo-lism.

We performed this research mainly to explore the effect of bone sialoprotein (BSP) silence by siRNA on the adhesion ability to bone matrix of bone-seeking breast cancer cells (MDA-MB-231BO). Also we aimed to provide experimental data for prevention and treatment of breast cancer bone metastasis by targeting BSP. We explored the effects of BSP gene silence on characteristics of bone-seeking breast cancer cells: proliferation by MTS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] assay, bone adhesion ability by a mouse bone adhesion model in vitro, morphology of the cells by SEM, and secretion of transforming growth factor-beta1 (TGF-beta1) and receptor activator of nuclear factor-kappa B ligand (RANKL) by ELISA kits. We performed intra-cardiac injection in nude mice to explore bone metastatic ability of different cell lines. The results showed that knockdown of BSP significantly inhibited the proliferation of MDA-MB-231BO cells and their adhesion to bone matrix. We also observed bone destruction caused by bone resorption around some adhering cells. The appearances of the cells changed in BSP gene silenced group, and the secretion of TGF-beta1 and RANKL decreased. The results showed BSP gene silence can partial inhibition bone metastasis of breast cancer cells in nude mice by X-ray assay and hematoxylin-eosin staining. Based on our research, siRNA-mediated BSP silencing can inhibit proliferation and adhesion to bone matrix of bone-seeking breast cancer cells and change their surface structure, thus inhibits their bone metastatic ability.
Animals ; Bone Matrix ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; Cell Adhesion ; Female ; Gene Silencing ; Humans ; Integrin-Binding Sialoprotein ; genetics ; pharmacology ; Mice ; Mice, Nude ; Neoplasm Metastasis ; genetics ; prevention & control ; RNA, Small Interfering ; genetics ; Tumor Cells, Cultured

OBJECTIVETo screen for the inhibitor of vascular endothelial growth factor (VEGF) 165 from random peptide library.

METHODSPositive phage clones were rescued after two rounds of panning and competitive elution. Its affinity activity to KDR was monitored through ELISA, immunohistochemical method, Chicken CAM assay and MTT.

RESULTSFive specific binding positive target molecule phage clones were obtained which were able to bind to cells whose surface had high KDR, among which, clone 3 and 13 could effectively block the vascularization of the chorioallantoic membrane of chick embryo, but they were not inhibitive on the proliferation of high KDR expression cells.

CONCLUSIONThe peptides, being the inhibitors of VEGF, may be useful in the treatment of cancers.

Animals ; Binding Sites ; Endothelial Growth Factors ; antagonists & inhibitors ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lymphokines ; antagonists & inhibitors ; metabolism ; Peptide Library ; Peptides ; pharmacology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo study the role of neuronal nitric oxide synthase (nNOS) in aged rats' hippocampal delayed neuronal death (DND) following brain ischemia.

METHODSModels of incomplete brain ischemia were induced by clipping common carotid artery. A total of 46 aged SD rats were divided into 8 groups: normal control group (Group A, n=5), sham-operation group (Group B, n=5), reperfusion 1, 6, 12, 24, 48, and 96 hours groups after brain ischemia for 30 minutes (Group C, D, E, F, G, and H, n=6/group). The expression of nNOS was examined by immunohistochemistry and neuronal ultrastructural changes were observed by the transmission electron microscopy (TEM) at different time points after reperfusion.

RESULTSImmunohistochemistry showed that nNOS expression in the hippocampal neurons was high in Group E, low expression in Group D, moderate expression in Group F and G. There was nearly no expression of nNOS in Group A, B, C, and H. Ultrastructure of hippocampal neurons was damaged more severely in reperfusion over 24 hours groups.

CONCLUSIONSNitric oxide (NO) may be one of the important factors in inducing DND after ischemia/reperfusion.

Animals ; Apoptosis ; Brain Ischemia ; enzymology ; Female ; Hippocampus ; enzymology ; pathology ; Immunohistochemistry ; Male ; Microscopy, Electron ; Neurons ; enzymology ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; enzymology

OBJECTIVE: To investigate if hyperbaric oxygen ( HBO) may induce structural changes of neurons in hippocampus from infantile rats and if the changes are reversible. METHODS: All 27 healthy SD infantile rats were exposed to HBO (0.25 MPa) or hyperbaric air (HBA) for 1 to 3 courses (10 days as 1 course). The hippocampus was taken at the end of each course to observe its morphology b y light microscope and electron microscope. RESULTS: HBO exposure induced capillary dilation, nuclear membr ane winding or blurring and some mitochondria swelling with its crista blurring i n neurons. The changes occurred after 1 course exposure and became significant w ith time. Most of the changes recovered 20 days after stopping exposure. No chan ge was found after HBA exposure. CONCLUSIONS: Long-term HBO exposure can cause capillary dilati on and ultrastructural injury of neurons in hippocampus from infantile rats. The damage is not serious, but reversible.