Background: High post-operative recurrence and poor prognosis are likely to be related to the infiltrative growth of the glioblastoma multiforme (GBM). Objectives: The primary objective of this study is to investigate the possible synergistic effect of the combined treatment of gamma knife radio-surgery (GKRS) and gene therapy for GBM and secondary objective is to explore the role of GKRS for the temporal and spatial regulation of the gene expression. Materials and Methods: The study performed on 70 nude mice and randomly divided into seven groups. Subcutaneous injection of human GBM tumor cells (T98G) was carried out to establish the animal models. Various doses of liposome-mediated pcDNA3.1-Egr. 1p-p16 recombinant plasmid were transfected through intra-tumor injection. GKRS was scheduled following the plasmid transfection. Tumor volumes were measured every 4 days after the treatment. Subcutaneous tumor nodule specimens were collected to analyze the cell apoptosis and p16 gene expression using terminal-deoxynucleoitidyl transferase mediated nick end labeling staining and reverse transcription-polymerase chain reaction. Tumor volumes, levels of cell apoptosis and p16 gene expression were compared between groups. Results: Rates of tumor growth were significantly lower in the pcDNA3.1-Egr. 1p-p16 plasmid + GKRS groups than that in the remaining groups 28 days following the GKRS management. The p16mRNA expression was noted in both of the pcDNA3.1-Egr. 1p-p16 plasmid group and the pcDNA3.1-Egr. 1p-p16 plasmid + GKRS with marginal-dose of 20 Gy group. The level of messenger ribonucleic acid expression was higher in the pcDNA3.1-Egr. 1p-p16 plasmid + GKRS with the marginal-dose of 20 Gy group, with a markedly increased apoptotic and necrotic cells, than that in the pcDNA3.1-Egr. 1p-p16 plasmid group. Conclusions: In animal studies, pcDNA3.1-Egr. 1p-p16 in combination with GKRS is a preferable management option for the GBM to the sole use of GKRS or gene therapy. It may be a novel approach for the treatment of human patient with GBM.
OBJECTIVE: To investigate the anti atherosclerosis effect and related mechanisms of total flavone of radix puerariae (TFRP) on atherosclerotic plaques in apoE gene deficiency (apoE-/-) mice. METHODS: apoE-/- mice were treated with saline, TFRP 15 mg . kg(-1). d(-1) or TFRP 85 mg . kg-1. d-1 (n = 8 each group) respectively per gavage for 12 weeks. The apoptotic cells in atherosclerotic plaques were then detected by TUNEL analysis, transmission electron microscope (TEM). The expression of CD-68, SMA and Caspase-3 were determined by immunochemical methods. RESULTS: Early macrophage apoptosis signs were observed under TEM, TUNEL-positive and CD-68 positive cells were found in lipid cores of atherosclerotic plaques. TFRP significantly reduced the number of apoptotic cells in a dose-dependent manner [(0.38 +/- 0.17)%, (1.95 +/- 1.02)%, (10.50 +/- 5.89)%, respectively, P < 0.01] in atherosclerotic plaques. TFRP treatment also significantly reduced the immune expression of Caspase-3 protein in a dose-dependent manner. CONCLUSION: TFRP significantly attenuated the development of advanced atherosclerotic plaques in a dose-dependent manner which might related to down-regulated expression of Caspase-3 protein and reduced macrophage apoptotic cells in atherosclerotic plaques post TFRP treatment.
Objective
To investigate the effect of umbilical cord blood monocytes(UCBMC) transplantation in neonatal rats with hypoxic-ischemic brain damage(HIBD) on rat astrocyte proliferation and its correlation with bone morphogenetic protein(BMP) 4.
Methods
Forty 7-day-old SD rats with the random number table, were divided into normal control(CON) group, hypoxic-ischemic(HI) group, normal(N)+ UCBMC group, HI+ UCBMC group, 10 rats in each group.HIBD model was prepared according to the Rice method.Twenty-four hours after hypoxia, the UCBMC group and HI+ UCBMC group were injected with 3×106 UCBMC via the lateral ventricle.Seven days after transplantation, changes in the number of neurons were observed by Nissl staining; the expression of glial fibrillary acidic protein(GFAP) was observed by Western blot; the astrocyte proliferation was observed by proliferating cell nuclear antigen(PCNA)/GFAP, BMP4/GFAP immunofluorescence double staining, and their correlation was analyzed.
Results
Nissl staining showed that the neurons at cerebral cortex and hippocamp were irregularly arranged and decreased in the HI group, and the number of Nissl-stained cells were significantly less than that in the CON group(tcortex=26.54, thippocamp=32.26, all P<0.05); but the Nissl-stained cells were well arranged in the HI+ UCBMC group and more Nissl-stained cells were observed as compared with the HI group(tcortex=10.18, thippocamp=12.56, all P<0.05); Western blot showed that the expression of GFAP in the HI group was significantly higher than that in CON group(t=5.50, P<0.05); but the expression of GFAP in the HI+ UCBMC group was significantly lower than that in the HI group(t=3.04, P<0.05); immunofluorescence double staining showed that there were more PCNA+ GFAP+ cells in the HI group compared with the CON group(t=10.39, P<0.05), but fewer PCNA+ GFAP+ cells were observed in the HI+ UCBMC group than those in the HI group(t=3.72, P<0.05). And there were more BMP4+ cells in the HI group compared with the CON group(t=5.52, P<0.05), but fewer BMP4+ cells were observed in the HI+ UCBMC group than those in the HI group(t=2.33, P<0.05). The fluorescence intensity of GFAP were correlated with that of BMP4 in HI+ UCBMC group(r=0.84, P<0.05).
Conclusions
UCBMC transplantation can decrease the proliferation of the astrocytes, thus promote brain damage repair and its mechanism might be collected with the decreased expression of BMP4.
Key words:
Umbilical cord blood monocytes; Hypoxic-ischemic brain damage; Astrocyte; Bone morphogenetic protein 4; Rat
Background:Astrocytes become reactive following many types of CNS injuries.Excessive astrogliosis is detrimental and contributes to neuronal damage.We sought to determine whether inhibition of cell cycle could decrease the proliferation of astroglial cells and therefore reduce excessive gliosis and glial scar formation after focal ischemia.Methods:Cerebral infarction model was induced by photothrombosis method.Rats were examined using MRI,and lesion volumes were estimated on day 3 post-infarction.The expression of glial fibrillary acidic protein(GFAP) and proliferating cell nuclear antigen(PCNA) was observed by immunofluorescence staining.Protein levels for GFAP,PCNA,Cyclin A and Cyclin B1 were determined by Western blot analysis from the ischemic and sham animals sacrificed at 3,7,30 days after operation.Results:Cell cycle inhibitor olomoucine significantly suppressed GFAP and PCNA expression and reduced lesion volume after cerebral ischemia.In parallel studies,we found dense astroglial scar in boundary zone of vehicle-treated rats at 7 and 30 days.Olomoucine can markedly attenuate astroglial scar formation.Western blot analysis showed increased protein levels of GFAP,PCNA,Cyclin A and Cyclin B1 after ischemia,which was reduced by olomoucine treatment.Conclusion: Our results suggested that astroglial activation,proliferation and subsequently astroglial scar formation could be partially inhibited by regulation of cell cycle.Cell cycle modulation thereby provides a potential promising strategy to treat cerebral ischemia.
In this paper, the ultrastructure of the Microsporidian is described in detail, such as the spore wall, the polar filament, polaroplast, anchoring disc, posterior vacuole and so on. By observing the ultrastructure of Penarus monodon infected by the Microsporidian, we give some clear ultrastructure photographs of the Microsporidian, and outline the symptom and precluding measure of the Microsporidian disease.
Objective To investigate bcl 2/bax expressions and then relationship with acute rejection in small intestine transplantation. Methods Inbred F344/N and Wistar/A rats were used to establish small intestinal transplantaion model. The recipients were designed to control, isograft, allograft and allograft plus FK506 treatment groups. The control group only received laporotomy. The expressions of bcl 2 and bax was detected by immunohistochemistry. Results The expression of bcl 2 in small intestine mucous membrane epithelium during acute rejection decreased remarkedly (P 0.05) on the third day postoperatively in allograft group, and further decreased (P 0.01) with time. There was no significant difference in bax expression between allograft group and the control group (P 0.05). There was no significant difference in bcl 2 and bax expression between FK506 treatment group and the control group (P 0.05). Conclusion High expression of bcl 2 is an early indicator for acute rejection of small intestinal allograft.
This study aimed to develop a specific and sensitive method for the rapid detection of NF-κB activity in pancreatic tissue. Male Wistar rats were randomly divided into two groups: (1) 16 rats in the acute pancreatitis (AP) group received retrograde injection of 5% sodium taurocholate (STC) into the biliopancreatic duct, and (2) 16 rats in the Control group received saline. NF-κB activation in rat pancreatic acinar cells was assessed by flow cytometry (FCM). We found that the NF-κB activity in the AP group significantly increased at 1.5 h (29.80%±7.83), had a peak at 3 h (65.17%±13.22), and then decreased gradually to 12 h time point, close to the level after 1.5 h stimulation of STC. The NF-κB activity of the Control group did not significantly vary at different time points (P>0.05). FCM is a specific and sensitive assay for the rapid detection of NF-κB activity in pancreatic tissue.
Strength and stiffness properties of materials are widely studied and used in civil engineering practice. However, most studies are based on unconfined conditions, which are different from real status of soil. This study investigated the primary yielding and yield locus for cement-stabilized marine clay. In this study, two types of cement-stabilized soils were studied through isotropic compression, triaxial drained shearing, unconfined compression, and bender element testing. Specimens with 20–50% of cement content and 7–90 days of curing period were used for the tests. Stress–strain behavior and primary yielding were evaluated, followed by construction of the primary yield locus. The characteristics of the primary yield locus and its development with curing time then were studied. The results showed that the properties of the primary yield locus were dependent on the type of stabilized soil, but were independent of the cement content and curing period. Thus, the approach provides a way to estimate the primary yield stress and drained stress path before primary yielding for cement-stabilized soil under confined condition. An empirical function was used to fit the primary yield locus. The primary isotropic yield stress was correlated to unconfined compressive strength or maximum shear modulus. Three indirect methods were proposed to predict the primary yield stress for cement-stabilized marine clay. The results showed that the primary yield stress can be estimated with reasonable accuracy.
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