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Objective To construct RGD-TAT modified liposomes(RGD-TAT-LPs)and evaluate its glioma targeting efficiency.Methods RGD-TAT-LPs was constructed by film-ultrasonic method,its appearance,particle size and Zeta potential were mearsured. Cellular uptake of LPs,TAT-LPs, RGD-LPs and RGD-TAT-LPs was used to evaluate the affinity to C6 cells.C6 cells were xenografted in athymic mice to establish the animal model,which were used to evaluate the distribution of liposomes in vivo. Results The particle diameter of RGD-TAT-LPs was (1 16.5 ±1 1.3 )nm,and its Zeta potential was (23.2 ±3.5 )mV. Cellular uptake experiments demonstrated the cell uptake efficiency of RGD-TAT-LPs by C6 cells were 2.9-fold,2.3-fold and 4.7-fold than that of RGD-LPs,TAT-LPs and LPs respectively. The in vivo imaging showed that RGD-TAT-LPs had the strongest fluorescence intensity in brain. Conclusion The RGD-TAT-LPs might serve as a promising delivery system of antitumor drugs.
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BACKGROUND:Rapid development of spermatogonial stem cells is the new hope for assisting reproductive technologies,and the stability of the number and structure of the chromosome cultured in vitro is one of the important factors in usage.OBJECTIVE: To study the influential factors on G-band exhibition in mouse spermatogonial stem cells, so as to offer a technology to identify karyotype of stem cells in culture.DESIGN: Observational experiment.SETTING: Luzhou Medical College.MATERIALS: The experiment was conducted in the Laboratory of Medical Molecular Biology, Luzhou Medical College from March 2004 to April 2005. 10 days old male and female Kunming mice were provided by Department of Animal of Luzhou Medical College (number of license No. 17 of experimental animal quality administration in Sichuan province).There were low-sugar DMEM medium, 10 mg/L Mitomycin-C, IMDM medium, 1 ×10-5 mol/L colchicines (PBS allocation),7.5 mmol/L KCL, fixative (mixture glacial acetic acid with methanol in 1:3), Giemsa staining solution and 0.25% zymine.METHODS: Bone marrow was aspirated from the thigh bone of mouse for feeder layer cells preparation. Cells from the male mice testis of 7-8 days after birth were prepared and made into cell suspension. After adjusting the cell density to 3×105 L-1, they were inoculated into the feeder layer of bone marrow stromal cells. Cells were cultured at 37 ℃ in CO2 incubator containing CO2 of 0.05 volume fraction and 70% humidity. The proliferation groups of stem cells cultured 15-20 days were selected, stirred and spread, and then treated with colchicine for 4-6 hours. Cell suspension was collected,and then stained after hypotonic treatment. The cells whose chromosomes were dispersed well and in metaphase were selected, and number of chromosomes was counted, and then the morphology of chromosomes was observed.MAIN OUTCOME MEASURES: Culture of bone marrow stromal cells and spermatogonial stem cells and coloration and count of chromosomes.RESULTS: The karyotype of spermatogonial stem cells was the same as the body cells of normal mouse, which showed granule or rod-shape. Karyotype was 20 pairs, 40 bars. Three kinds of chromosome morphology could be observed under oil immersion lens. The first type was condensed, which could be counted in total, but the band could not be seen.The second type was chromatosome that spread completely in the center of equatorial plate and were in metaphase. In this phase, total numbers and band were seen clearly. Last type-chromosomes had already folded and moved towards two poles and concentrated gradually, the total number of chromosomes could be counted, but bending could not be seen clearly.CONCLUSION:Many factors can affect the karyotype of spermatogonial stem cells, including the phase of cell division,effect of hypotonic solution, diffusion of the cell when dropping slides, concentration of trypsin and digestion time, etc.
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BACKGROUND: The c-fos gene is commonly expressed in neurons,which may act as one of the signs of activities and reflection of injured neural cells.OBJECTIVE: To investigate the expression of hippocampal c-fos gene of rats with ischemic cerebral injury and the protective role of shenmai injection at the molecular level.DESIGN: Randomized controlled study.SETTING: Department of Histology and Embryology, Luzhou Medical College.MATERIALS: The experiment was completed at the Department of Histology and Embryology, Luzhou Medical College from January to March 2002. Totally 30 male Wistar rats were randomly divided into control group, ischemia group and treatment group with 10 in each group.METHODS: Bilateral common carotid arteries of rats in the ischemia and treatment groups were splinted for 30 minutes and reperfused for 1 hour to establish models of ischemic cerebral injury. Rats in the treatment group were injected with 2 mL/kg of shenmai (constituted with hongshen, dwarf lilyturf tuber and other Chinese herbal medicines) 30 minutes before ischemia. Rats in the ischemia group were not treated with any drugs, and rats in the control group were treated with sham operation but without splinting common carotid artery and giving any drugs. Hippocampal tissue of rats was obtained to make paraffin sections. In every group, one section from each rat was taken at random. Totally 100 neurons of every group were counted. Expression of c-fos gene in hippocampal neuron was observed according to the nuclear color of neurons (+++ as dark-brown mark;++ as brown mark; + as light brown mark; - as no brown mark).MAIN OUTCOME MEASURE: c-fos expression of hippocampal neurons of rats in each group.RESULTS: Totally 30 rats entered the final analysis. Expression of c-fos gene in hippocampal neurons was significantly more in the ischemia group than that in the control group (+++: 24/visions, 7/visions, P ＜ 0.05); but that in the treatment group was less than that in the ischemia group (+++:13/visions, 24/visions, P ＜ 0.05).CONCLUSION: Shenmai injection can reduce the expression of c-fos gene in hippocampal neurons of rats with ischemic cerebral injury, and can protect neural cells of ischemic cerebral injured tissue.
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Objective To evaluate the diagnostic value of the ultrasonography in greater omentum lesions.Methods The ultrasonic findings in 33 patients with greater omentum lesions were compared with the pat hological result of operation,celioscope or ascites cytology.Results The lesions in 27 of 33 patients(81.8%)were localed within the treater omemurn,and pathological changes of 26/33(78.8%)were found by ultrasonography.These greater omentum lesions appeared as characteristics of thickened,shifted and peculiar-echo.Conclusion The lOcation and pathological nature of the primary affection can be approximately estinlated according to the localizatiOn and sonographic appearance of the greater omentum by experienced physicians.