Therapeutic effect of ovarian intra-arterial infusion of GE7-delivery system-mediated HSVl-tk/ganciclovir gene therapy in a rat model of malignant ovarian tumor / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the gene expression of herpes simplex virus type 1 thymidine kinase (HSVl-tk) in rat malignant ovarian tumor tissues and the therapeutic effect of ganciclovior (GCV) after intra-arterial infusion of HSVl-tk gene therapy mediated by GE7-delivery system.</p><p><b>METHODS</b>A GE7-polylysine/pCMV-HSV1-tk/polylysine-HA20 4-element complex was constructed. Eighteen rats with DMBA-induced ovarian tumor were divided into 3 groups as Atk, ANS and Vtk groups. The 4-element complex GE7-polylysine/pCMV-HSV1-tk/polylysine-HA20 was injected via the ovarian artery into the rats of Atk group, saline buffer was injected in the ANS groups, and the 4-element complex was injected via the tail vein into the rats of Vtk group. All rats received intraperitoneal injection of GCV in a dose of 50 mg/kg daily for 10 days. The rats were sacrificed 3 days after the final dose of GCV, and the tumor weight was measured and tumor growth inhibition rate was calculated. Flow cytometry was used to assess the cell cycle and apoptosis.</p><p><b>RESULTS</b>The tumor weight in the rats of Atk group was (4.77 ± 2.31) g, significantly lower than that of ANS group [(14.66 ± 6.26) g, P < 0.01] and Vtk group [(17.53 ± 7.19) g, P < 0.01]. The tumor growth inhibition rate of the Atk group was 67.5%, while that of Vtk group was -19.6%. The flow cytometry showed that S-phase tumor cells in the Atk group were (54.32 ± 9.65)%, significantly higher than that in the ANS (27.43 ± 9.22)% and (30.16 ± 11.57)% in the Vtk group (both P < 0.01). The tumor cell apoptosis rate in the Atk group was (39.15 ± 12.16)%, significantly higher than that in the ANS group [(11.86 ± 5.28)%, P < 0.01] and Vtk group [(14.32 ± 6.43)%, P < 0.01].</p><p><b>CONCLUSION</b>HSV1-tk/GCV gene therapy system mediated by GE7 non-viral delivery system via ovarian arterial infusion effectively causes cell cycle arrest at S phase and enhances cell apoptosis, therefore, exerts an inhibitory effect on tumor growth.</p>

Evaluation of GE7-transferring system-mediated HSV(1)-tk gene transfer in a rat model of ovarian tumor via intra-arterial route / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the gene and protein expression of herpes simplex virus type I-thymidine kinase (HSV(1)-tk) in the ovarian tumor tissues and other organs after arterial infusion of HSV(1)-tk gene mediated by GE7 delivery system.</p><p><b>METHODS</b>GE7-polylysine/pCMV-HSV(1)-tk/polylysine-HA20 complexes were constructed. Nine rats with induced ovarian tumor were divided into 3 groups, injecting the 4-element complexes or saline buffer through the ovarian artery and complexes through the tail vein, respectively. The ovarian tumors, hearts, livers, spleens, lungs and kidneys were obtained at 72 hours after injection. RT-PCR and Western Blot were preceeded to determine the expression of HSV(1)-tk gene and protein in the tumor tissues and other organs.</p><p><b>RESULTS</b>In the group of arterial injection with 4-element complexes, the HSV(1)-tk gene and protein were expressed strongly in the tumor tissues, while little or none was detected in other organs. In the group of arterial injection with saline buffer, no HSV(1)-tk gene and protein was detected in both tumor tissues and other organs. In the group of tail vein injection, none was detected in tumor tissues and only little was found in the livers, spleens, lungs and kidneys.</p><p><b>CONCLUSION</b>High target and gene transfer rates can be obtained when HSV(1)-tk gene is transferred via the artery route mediated by GE7 delivery system. HSV(1)-tk protein can be expressed after the gene transfer. The results may provide a new strategy for target killing of HSV(1)-tk/GCV system in ovarian tumors.</p>

Effect of Gengnianchun Recipe on learning memory function and hippocampal cholinergic system in ovariectomized rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the effect of Gengnianchun Recipe (GNC) on learning memory function and its regulatory effect on hippocampal cholinergic system in ovariectomized rats.</p><p><b>METHODS</b>Female rats 10-12 months old were randomized into 5 groups, the sham-operation group, the model group treated with normal saline, the positive control group treated with Nilestriol, and the two GNC groups treated with high and low dose GNC respectively. A little fat around ovary was cut in the sham-operation group. The treatment lasted for 12 weeks after ovariectomy. Changes of learning memory function were tested by Morris water maze; serum level of estradiol (E2) was measured by chemical fluorescent method; hippocampal acetylcholinesterase (AChE) mRNA was determined with Real-time PCR; and the activities of acetylcholine (ACh), AChE and choline acetyltransferase (ChAT) in hippocampus were detected by immunohistochemistry respectively.</p><p><b>RESULTS</b>Twelve weeks after ovariectomy, serum E2 and learning memory function markedly decreased in the ovariectomized rats (P < 0.01, P < 0.05). Nilestriol and high dose GNC showed an effect in improving the symptoms of learning memory functional deprivation and elevating the activities of hippocampal ACh, AChE and ChAT (all P < 0.05).</p><p><b>CONCLUSION</b>GNC can improve learning memory function of ovariectomized rats, and its mechanism might be realized by regulating the cholinergic system in hippocampus.</p>

Proteomic analysis of plasma membrane of HBV transgene mice livers / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study liver plasma membrane (PM) proteins affected by hepatitis B virus (HBV), and to study the molecular mechanism of HBV infection through plasma membrane proteome analysis of transgene mice livers.</p><p><b>METHODS</b>Plasma membrane was purified using second antibody superparamagnetic beads that combine subcellular fractionation and immunoisolation strategies. Western blot was used to verify the purification of plasma membrane and the expression of HBV envelope protein. The proteins from plasma membrane were extracted, quantified and separated by two-dimensional electrophoresis. The gels were stained by silver nitrate or Coomassie Blue and analyzed by Imagemaster software. The different expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF-TOF MS).</p><p><b>RESULTS</b>The plasma membrane of mice livers was enriched 3-fold and the contamination from mitochondria was reduced 2-fold in comparison with the density gradient centrifugation method. HBV envelope protein was found only in HBV transgene mice livers. More than 500 protein spots were separated in Coomassie Blue stained gels. Twenty-six proteins with two-fold differences were found through Imagemaster software analysis. Seven proteins were identified.</p><p><b>CONCLUSION</b>An optimized plasma membrane purification method was developed; protein profile of liver plasma membrane changed in the transgene mice livers; some proteins related to HBV infection were found, and this work may be helpful in the research of the molecular mechanism of HBV infection.</p>

Effect of Gengnianchun Recipe on bone mineral density, bone biomechanical parameters and serum lipid level in ovariectomized rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effect of Gengnianchun Recipe (GNC) on bone mineral density (BMD), bone biomechanical parameters and serum lipid level in the bilaterally ovariectomized (OVX) rats and to explore the prophylactic and therapeutic action of GNC on ovariectomy induced osteoporosis and hyperlipidemia.</p><p><b>METHODS</b>OVX SD rats, 10 - 12 months old, were divided into different groups and fed with GNC 2 g/d, GNC 1 g/d and Nilestriol 0.125 mg/week, respectively for 4 months to observe the change of BMD and bone biomechanical parameters of the lumbar vertebrae, and the serum levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C), and to compare the effect of the two drugs on the morphology of the uterus.</p><p><b>RESULTS</b>There was marked reduction in BMD and biomechanical parameters in lumbar vertebrae (P < 0.01) and increase of serum TC and LDL-C levels (P < 0.01) in rats after OVX. GNC or Nilestriol significantly improved the decreased BMD and biomechanical parameters of the lumbar vertebrae (P < 0.05 or P < 0.01), and reduced the serum TC and LDL-C levels (P < 0.01). In the Nilestriol group, the wet weight of uterus got increased obviously (P < 0.01), the number of uterine glands increased, uterine columnar epithelium thickened, and the mitotic figures in the epithelial stroma and myointimal cells augmented. But no such effect in wet weight and morphology of uterus was found in the GNC group.</p><p><b>CONCLUSION</b>GNC could increase the BMD and biomechanical parameters of the lumbar vertebrae, reduce the serum TC and LDL-C levels, yet produce no adverse reaction in stimulating proliferation and hypertrophy of uterus.</p>