Objective To investigate the influence of cationic liposomemediated endostatin gene on colorectal cancer liver metastasis. Methods Animal model for colorectal carcinoma liver metastasis were established. The plasmid expressing endostatin genelipofectAMINE were injected in vein. Results After cationic liposomemediated endostatin gene were injected in vein, the incidence of liver metastasis and mean numbers of liver tumors were decreased, survival time of animal was significantly longer. Conclusion Intravenous injection of cationic liposomemediated endostatin gene can control the development of colorectal cancer liver metastasis effectively.
【Abstract】ObjectiveTo construct a recombinant eukaryotic expression vector for human endostatin in order to study the inhibitory effect of liposome-mediated endostatin gene on the growth of human liver carcinoma in nude mice. MethodsHuman endostatin cDNA including IL-2 secreting peptide was cloned into eukaryotic expression plasmid pcDNA3.0 to construct recombinant plasmid pCD-sEndo. pCD-sEndo plasmid was transferred into hepatocarcinoma cell line SMMC-7721 mediated by liposome Dosper, the expression and secretion of endostatin gene was detected by RTPCR and Western blot analysis. The suspension of SMMC-7721 cells was injected subcutaneously at the back of 32 nude mice to establish the model of human liver carcinoma. The mice were divided into 4 groups randomly, and injected with Dosper+pCD-sEndo, Dosper+pcDNA3.0, Dosper and physiological brine separately. Tumor volume was measured by stages. The mice were executed after the drug had been given for 1 week, then the microvessel density (MVD) of the tumor tissue was detected with immunohistochemical method and apoptotic index of tumor cells was measured by TUNEL-stain. ResultsThe eukaryotic expression vector pCD-sEndo was successfully constructed and was confirmed by enzyme digestion and sequence analysis. Expression of endostatin gene was detected in transfected SMMS-7721 cells by RTPCR in vitro, and endostatin protein was also detected in the supernatant of transfected SMMS-7721 cells by Western blot. In vivo study, the growth of human liver carcinoma was inhibited in the group injected with endostatin gene: the average volume of tumor in this group was significantly smaller than that in other groups （P＜0.05）; the average MVD in this group was 6.2±2.5, significantly less than that in the group injected with physiological brine (32.8±6.4), Dosper （27.8±6.4）, or Dosper+pcDNA3.0 (25.5±5.5), P＜0.05. The average apoptotic index of tumor cells in treatment group, brine group, Dosper group and Desper+pcDNA3.0 group was 24.5±7.3, 7.6±2.5, 9.5±3.0 and 11.2±3.6 respectively, it was evidently higher in the treatment group than in the latter three groups. ConclusionHuman endostatin mediated by cation liposome could decrease the microvessel number of implanted human hepatocarcinoma in nude mice. It could also accelerate apoptosis of tumor cells and inhibit growth of tumor.
Liposome is an ideal drug carrier with many advantages such as excellent biocompatibility, non-immunogenicity, and easy functionalization, and has been used for the clinical treatment of many diseases including tumors. For the treatment of tumors, liposome has some passive targeting capability, but the passive targeting effect alone is very limited in improving the drug enrichment in tumor tissues, and active targeting is an effective strategy to improve the drug enrichment. Therefore, active targeting liposome drug-carriers have been extensively studied for decades. In this paper, we review the research progresses on active targeting liposome drug-carriers based on the specific binding of the carriers to the surface of tumor cells, and summarize the opportunities, challenges and future prospects in this field.
ObjectiveTo evaluate the therapeutic effect and adverse reaction of paclitaxel liposome combined with continuous infusion of large-dose 5-fluorouracil(5-fu) in treatment for advance gastric cancer(AGC). MethodsFrom May 2009 to August 2012, 63 consecutive patients with AGC in this hospital were enrolled in this study. All the patients were given chemotherapy including paclitaxel liposome and continuous infusion of large-dose(2.5 g/m2) 5-fu. The efficacy and toxicity of this regimen were observed. ResultsThere was no patient who could not tolerate adverse reaction related to such regimen. Five cases achieved complete response and 31 cases achieved partial response, the overall response rate was 57.1%(36/63). Hematologic toxicity included gradeⅢ/Ⅳleucopenia 8 cases(12.7%) and neutropenia 10 cases(15.9%), while there was no occurrence of gradeⅢ/Ⅳanemia or thrombopenia. Non-hematologic toxicity was fairly mild. ConclusionsPaclitaxel liposome is safe, well tolerated, highly targeted, and has long duration of effect. Paclitaxel liposome combined with continuous infusion of large-dose 5-fu is safe and effective in treatment for patients with AGC.
OBJECTIVE: To study the therapeutic effect of Suyuping in which liposome is the main constituent on II degree burn wound. METHODS: From October 1998 to October 1999, 42 cases with II degree burn wound were adopted in this study. Among them, there were 30 males and 12 females, the average wound area was (23.4 +/- 9.7)%. The wounds on the left side of body were managed with sulfadiazine argentum(SD-Ag) while that on the right side with Suyuping. Healing and other situation about the burn wounds were observed and recorded at times. RESULTS: Forty-two patients completely healed when discharged from hospital. The average wound area managed with Suyuping was (11.2 +/- 7.3)% and the area with SD-Ag was (9.4 +/- 5.8)%. The mean healing time of Suyuping group was(18.4 +/- 4.7) days while that of SD-Ag was (23.5 +/- 7.9) days, there was significant difference(P lt; 0.05). Suyuping group presented few wound pain, allergy, bleeding and the side effect was less. CONCLUSION: Suyuping can accelerate wound healing and attenuate wound pain, allergy and bleeding, it is a potential and specific topical agent for treating II degree burn wound.
Liposomes with precisely controlled composition are usually used as membrane model systems to investigate the fundamental interactions of membrane components under well-defined conditions. Hydration method is the most common method for liposome formation which is found to be influenced by composition of the medium. In this paper, the effects of small alcohol (ethanol) on the hydration of lipid molecules and the formation of liposomes were investigated, as well as its coexistence with sodium chloride. It was found that ethanol showed the opposite effect to that of sodium chloride on the hydration of lipid molecules and the formation of liposomes. The presence of ethanol promoted the formation of liposomes within a certain range of ethanol content, but that of sodium chloride suppressed the liposome formation. By investigating the fluorescence intensity and continuity of the swelled membranes as a function of contents of ethanol and sodium chloride, it was found that sodium chloride and ethanol showed the additive effect on the hydration of lipid molecules when they coexisted in the medium. The results may provide some reference for the efficient preparation of liposomes.
Microvesicles (MVs) is small membrane vesicles released from different cell types under different conditions. Studies have shown that MVs may mediate vascular inflammation, angiogenesis, and other pathological processes. MVs may play an important role in the pathogenesis of diabetic retinopathy (DR) by mediating endothelial cell injury, thrombosis and neovascularization. The plasma MV level may be an effective parameter to monitor the development of DR. This article will summarize the research progress of the relationship between MVs and DR in recent years.
Objective To investigate the inhibitory effect of proliferation cell nuclear antigen (PCNA) antisense oligonucleotides mediated by liposome transfection on hepatocellular carcinoma cell proliferation. MethodsThe antisense oligonucleotides were complementary to 18mer sequences next to the start codon of PCNA mRNA sequences. The human hepatocellular carcinoma cell line Bel7404 was treated with antisense oligonucleotides. The inhibition of proliferation was estimated by MTT method. We compared the deference between the liposome mediated transfection technique and direct transfection technique. ResultsThe cell proliferation was inhibited effectively by antisense oligonucleotides. A sense sequence oligomer showed no effect.Liposome mediated transfection could enhance the inhibitory effect. Conclusion Liposome mediated transfection could enhance the inhibitory effect of PCNA antisense oligonucleotides on hepatocellular carcinoma cell proliferation.
【Abstract】ObjectiveTo investigate the effect of genetic modulation of the hepatic graft with IL-10 during liver preservation in rat liver transplantation. MethodsEleven cases of orthotopic liver transplantation were performed in Lewis to BN rats according to the cuff’s technique. All rats were divided into 3 groups,which were control group(n=3), Lipo group(n=4) and Lipo-rIL-10 group(n=4). Lipofectamine 2000-pCR3.1 complex and Lipofectamine 2000-pCR3.1 rIL10 complex were respectively injected into portal vein and kept for 45 minutes to transfect grafts during cold preservation in vitro. All rats were killed on postoperative day 6. Serum samples were collected for decting IL-10 by means ELISA. Transgene expression of rIL-10 was assessed by means of RT-PCR and immunohistochemistry. ResultsIn Lipo-rIL-10 group, levels of IL-10 from suprahepatic vena cava were significant higher than those from infrahepatic vena cava (P=0.024), transgene expression of rIL-10 in Lipo-rIL-10 were higher than those of control group and Lipo group assessed by means of RT-PCR and immunohistochemistry. ConclusionDuring cold preservation in vitro through portal vein injection to donor liver, liposome mediated gene transfection can successfully achieve local gene expression.
Objective To observe the inhibition of LipofectamineTM2000 (LF2000)mediated pSUPER recombinant plasmid expressing small interference RNA targeting hypoxia-induced factor (HIF)-1alpha;(pSUPERsiHIF-1alpha;) on retinal neovascularization in mice. Methods pSUPERsiHIF-1alpha; recombinant plasmid was created. Forty-eight (seven-day-old) C57BL/6J mice were randomly divided into a normal group, the control group, empty vector group and gene therapy group with 12 mice in each group. Mice in the normal group were kept in normal room air, while the other three groups retinal neovascularization was induced by hypoxia. The mice in control group were not treated. The mice in the vector group received intravitreous injection of pSUPER and LF2000 (1 mu;l), and the gene therapy group received pSUPERsiHIF-1alpha; and LF2000 (1 mu;l)one day before being returned to normal room air．Fluorescent angiography was used to assess the vascular pattern. The proliferative neovascular response was quantified by counting the nuclei of new vessels extending from the retina into the vitreous in cross-sections．HIF-lalpha;and vascular endothelial growth factor (VEGF) levels in retinas were measured by immune histochemical staining method and reverse transeriptase-polymerase chain reaction (RT-PCR). Results Fluorescent angiography showed radial branching pattern vessels in the normal group and distorted large vessels, obstructed capillaries, many neovascular tuffs, fluorescence leakage in the peripheral retina in the control group and vector group. The gene therapy group demonstrated a significant reduction in neovascular tufts and fluorescence leakage compared with the control group and the vector group. The number of vascular cell nuclei extending breaking through the internal limiting membrane(ILM) of control group and vector group increased significantly compared with normal group (F=5850.016,P＜0.05), while obviously decreasing in the gene therapy group compared with control group (F=3012.469,P＜0.05). Immunohistochemical staining showed the expression of HIF-1alpha; protein in nucleus and VEGF protein in cytoplasm. The expression of HIF-1alpha; protein in retina was negative, while VEGF protein was weakly positive in normal group. The expression of HIF-1alpha; and VEGF protein were both positive in control group and vector group, while weakly positive in gene therapy group. The Results of RT-PCR showed that the expression of HIF-1alpha; mRNA in retina was increased significantly in control group and vector group as compared with normal group (F=3102.326,P＜0.05), while decreasing significantly in gene therapy group as compared with control group (F=3336.425,P＜0.05). Conclusion Retinal neovascularization in the mice is significantly inhibited by intravitreal injection of LF2000-mediated recombinant plasmid pSUPERsiHIF-1alpha;.