Objective:To explore the significance of elevating cervical flap before vital-dye injection in identifying blue-stained nodes receiving lymphatic drainage in rabbit's tongue, so as to provide raw data for normalizing vital-dye injection in sentinel lymph node biopsy (SLNB) during tongue-carcinoma treatment. Methods: 64 adult rabbits were randomly divided into four groups according to the injecting sites: tongue-tip, tongue-lateral, tongue-center and tongue-root. Four doses of methylene blue-0.1, 0.2, 0.3 and 0.4 ml were chosen and injected into 4 tongues in every group, 2 of them were injected before and after the elevation of cervical flap respectively. Time was recorded according to the time points of colored and discolored lymphatic duct and nodes. Results:①The overall success rate in localizing blue-stained node was 93.75%(60/64). ②In the success cases in group elevating cervical flap just before injecting methylene blue, all experiments had the time records of lymphatics and nodes' turning blue; while the counterpart group did not show any blue staining. ③All the blue stained nodes were found in lymph nodeⅠto Ⅲ local area. Conclusion:t may be dependable to utilize methylene blue as tracer in the tongue carcinoma SLNB. Elevation of cervical flap before injecting methylene blue is necessary and practicable in tongue carcinoma SLNB.

BACKGROUND: Recent researches indicate that ischemia and hypoxia can lead to abnormal brain metabolism and even energy failure, which is an important reason for brain damage and necrosis and identifies energy metabolism disorder as the key event in brain ischemia-reperfusion (IR)injury. Glucose transporter-3 plays the vital role in brain energy metabolism.OBJECTIVE: To observe the changes of cerebral infarct volume and glucose transporter-3 mRNA and protein expressions in cerebral cortical penumbra at different stages of focal cerebral ischemia and reperfusion in rats.DESIGN: Randomized controlled experiment.SETTING: Department of Neurosurgery, Second Hospital Affiliated to Sun Yat-sen University.MATERIALS: This experiment was conducted in the Animal Laboratory of Medical Research Center, Second Hospital Affiliated to Sun Yat-sen University between August and October 2002.Totally 56 SD rats were randomized into 3 groups which were subjected to ① ischemia for 1 hour followed by reperfusion (n=28), ② ischemia for 3 hours followed by reperfusion (n=24), and ③ sham operation (n=4). The rats in the first group were subdivided into 7 subgroups for examination at 1, 3, 6, 12, 24, and 72hours and 1 week after ischemia, with 7 rats in each subgroup; the rats in the second ischemia group were also subdivided in similar manner but without a 1 hour postischemic subgroup. The rats in the sham operation group only received the operation but without arterial occlusion.METHODS: Focal cerebral ischemia-reperfusion (IR) injury model was induced in the rats in the two ischemic groups by means of insertion of suture for arterial occlusion, and the ratio of central ischemic area to cerebral infarct volume in the ischemic penumbra was examined at the specified time points. Reverse transcription-PCR (RT-PCR) was used to detect the expression of glucose transporter-3 mRNA in the cerebral cortex in ischemic penumbra region, and semi-quantitative immunohistochemistry (IHC) employed to detect the level of glucose transporter-3 protein.MAIN OUTCOME MEASURES: Cerebral infarct volume after IR injury, changes of transporter-3 mRNA and protein expressions after IR injury.RESULTS: Totally 56 rats were used in this experiment and all entered result analysis. The post-IR cerebral infarct volume was obviously smaller in 1-hour ischemia group than in 3-hour ischemia group. Glucose transporter-3 mRNA expression began to increase 3 hours after ischemia in 1-hour ischemia group, reaching the peak level at 24 hours and still mainrained higher level than that of the sham operation 1 week; in 3-hour ischemia group, the mRNA expression was slightly decreased at 3 hours but began to increase afterwards till reaching the peak level at 24 hours, followed then by recovery of normal level at 1 week. The changes in glucose transporter-3 protein and mRNA expressions followed almost the same pattern.CONCLUSION: Glucose transporter-3 expression is up-regulated in the ischemic penumbra region, possibly as a protective response to cerebral IR injury.

AIM: To verify the role of enhancing or suppressing the expression of glutathione peroxidase 1 (GPx1) in the growth, migration and invasion of glioblastoma multiforme cell lines U87MG and U118MG.METHODS:U87MG and U118MG cell lines were transfected with the vector containing specific siRNA or pcDNA3.1 recombinant plas-mid both targeting GPx1.The mRNA and protein expression levels of GPx1 were detected by real-time PCR and Western blotting.MTS assay was applied for determining the cell activity.The abilities of migration and invasion were examined by Transwell assay.RESULTS:Compared with blank control group and negative group, the inhibitory rate of the cell activity in U87MG cells in siRNA group was significantly reduced by 25.9%, 35.7%and 34.8%at 24 h, 48 h and 72 h, respec-tively (P<0.05).In contrast, the cell activity of U118MG cells in pcDNA3.1-GPx1 group was significantly increased by 22.7%, 45.8%and 39.8%at 24 h, 48 h and 72 h, respectively ( P<0.05) .In siRNA group, the inhibitory rate of mi-gration in U87MG cells was 41.6%±8.2%and the invasion was 41.6%±8.2%compared with blank control group and negative group (P<0.05).The cell migration and invasion rates of the U118MG cells in pcDNA-GPx1 group were in-creased by 55.8%±9.8% and 60.8% ±9.2%, respectively, compared with blank control group and negative group (P<0.05).CONCLUSION:The down-regulation of GPx1 by specific siRNA reduces the capability of cell growth, mi-gration and invasion of U87MG cells, while up-regulation of GPx1 by pcDNA3.1-GPx1 increases the capability of cell growth, migration and invasion of U118MG cells.

AIM: To investigate the volume percentage of infarct and expression of glucose transporter-1 (GLUT1) transcription and protein levels at different ischemic time point and different reperfusion time point in rat focal cerebral ischemic penumbra. METHODS: Focal ischemic models of middle cerebral artery occlusion (MCAO) in rats were made by inserting nylon thread. Brain samples were harvested from ischemic penumbra. Infarct volume were analyzed quantitively by Kontron IBAS 2.5 image auto-analyses system. The expressien of GLUT1 mRNA was assessed by RT-PCR, and the expression of GLUT1 protein was assessed by immunohistochemistry. RESULTS: The infarction volume in MCAO 1 h/reperfusion (R) group was obviously smaller than that in MCAO 3 h/R group. GLUT1 increased at (1 h) MCAO 1 h/R group, climbed to climax at 24 h and remained higher than normal at 1 week. In contrast, in the MCAO 3 h/R group, the corresponding index was at 3 h, 24 h and 1 week, but the increasing degree of GLUT1 was slighter than MCAO 1 h/R. GLUT1 protein began to ascend at 1 h, reached climax at 24 h and was higher than normal at 1 week in MCAO 1 h/R group, while in MCAO 3 h/R group, the corresponding index was at 3 h, 24 h and 1 week. CONCLUSION: GLUT1 expression is notably up-regulated in the penumbra region after focal cerebral ischemia, it may be a protective reaction against ischemic injury. [

AIM: To investigate the volume percentage of infarct and expression level of glucose transporter-3 (GLUT3) transcription and protein at different ischemic time points and different reperfusion time points in rat focal cerebral ischemic penumbra. METHODS: Focal ischemic models of middle cerebral artery occlusion (MCAO) in rats were made by inserting nylon thread. Brain samples were harvested from ischemic penumbra. Infarct volume was analyzed quantitatively by Kontron IBAS 2.5 image auto-analyses system. The change of GLUT3 mRNA was assessed by RT-PCR, and the expression of GLUT3 protein was assessed by immunohistochemistry. RESULTS: The infarction volume in MCAO 1 h/R group was obviously smaller than that in MCAO 3 h/R group. GLUT3 began to ascend at 3 h in MCAO 1 h/R group, reached to climax at 24 h and remained higher than normal at 1 week. In contrast, in the MCAO 3 h/R group, GLUT3 had a descent at 3 h. Later on, it ascended rapidly, and reached climax at 24 h. At 1 week, it approached to normal. The expression level of GLUT3 protein corresponds with that of mRNA. CONCLUSION: GLUT3 expression is up-regulated in the penumbra region after focal cerebral ischemia, it may be a protective reaction against ischemia/reperfusion injury. [

This study established superparamagnetic iron oxide (SPIO)-labeled nerve growth factor-beta (NGF-beta) gene-modified spinal cord-derived neural stem cells (NSCs). The E14 rat embryonic spinal cord-derived NSCs were isolated and cultured. The cells of the third passage were transfected with plasmid pcDNA3-hNGFbeta by using FuGENE HD transfection reagent. The expression of NGF-beta was measured by immunocytochemistry and Western blotting. The positive clones were selected, allowed to proliferate and then labeled with SPIO, which was mediated by FuGENE HD transfection reagent. Prussian blue staining and transmission electron microscopy (TEM) were used to identify the SPIO particles in the cells. The distinctive markers for stem cells (nestin), neuron (beta-III-tubulin), oligodendrocyte (CNPase) and astrocyte (GFAP) were employed to evaluate the differentiation ability of the labeled cells. The immunocytochemistry and western blotting showed that NGF-beta was expressed in spinal cord-derived NSCs. Prussian blue staining indicated that numerous blue-stained particles appeared in the cytoplasma of the labeled cells. TEM showed that SPIO particles were found in vacuolar structures of different sizes and the cytoplasma. The immunocytochemistry demonstrated that the labeled cells were nestin-positive. After differentiation, the cells expressed beta-III-tubulin, CNPase and GFAP. It was concluded that the SPIO-labeled NGF-beta gene-modified spinal cord-derived NSC were successfully established, which are multipotent and capable of self-renewal.
Cells, Cultured ; Dextrans/*diagnostic use ; Embryo, Mammalian ; Magnetic Resonance Imaging ; Magnetics ; Magnetite Nanoparticles/*diagnostic use ; Nerve Growth Factor/*genetics ; Nerve Growth Factor/pharmacology ; Neural Stem Cells/*cytology ; Spinal Cord/*cytology ; Transfection

AIM:To study the effects of neurological improvement and remyelination after intracranial hemorrhage(ICH)in rats by a novel therapeutic strategy with hepatocyte growth factor(HGF)gene transfected human umbilical cord mesenchymal stem cells(hUCMSCs)by lentiviral vector.METHODS:ICH was induced in 60 adult male Sprague-Dawley rats by a stereotactically guided injection of bacterial type IV collagenase into the right internal capsule.Non-modified hUCMSCs,HGF-modified hUCMSCs with lentiviral vector or PBS were administered left intraventricularly 7 d after right internal capsule ICH.All rats underwent modified neurological severity scores for 35 d.Luxol fast blue staining,immunohistological staining and Western blotting assessments for myelin basic protein(MBP)were applied.RESULTS:The ICH rats receiving HGF-modified hUCMSCs demonstrated significant functional recovery,determined by modified neurological severity scores,compared to the other groups from 2 weeks after cell therapy.As indicated by Luxol fast blue staining,the percent area of demyelination was obviously reduced in the HGF-hUCMSC treatment group compared to the PBS control group and hUCMSC-only treatment group at 5 weeks after ICH.The expression of MBP detected by immunohistological staining and Western blotting was significantly higher in HGF-hUCMSCs treated brain than that in other groups(P

This study established superparamagnetic iron oxide (SPIO)-labeled nerve growth factor-β (NGF-β) gene-modified spinal cord-derived neural stem cells (NSCs).The E14 rat embryonic spinal cord-derived NSCs were isolated and cultured.The cells of the third passage were transfected with plasmid pcDNA3-hNGFβ by using FuGENE HD transfection reagent.The expression of NGF-β was measured by immunocytochemistry and Western blotting.The positive clones were selected,allowed to proliferate and then labeled with SPIO,which was mediated by FuGENE HD transfection reagent.Prussian blue staining and transmission electron microscopy (TEM) were used to identify the SPIO particles in the cells.The distinctive markers for stem cells (nestin),neuron (β-Ⅲ-tubulin),oligodendrocyte (CNPase) and astrocyte (GFAP) were employed to evaluate the differentiation ability of the labeled cells.The immunocytochemistry and western blotting showed that NGF-β was expressed in spinal cord-derived NSCs.Prussian blue staining indicated that numerous blue-stained particles appeared in the cytoplasma of the labeled cells.TEM showed that SPIO particles were found in vacuolar structures of different sizes and the cytoplasma.The immunocytochemistry demonstrated that the labeled cells were nestin-positive.After differentiation,the cells expressed β-Ⅲ-tubulin,CNPase and GFAE It was concluded that the SPIO-labeled NGF-β gene-modified spinal cord-derived NSC were successfully established,which are multipotent and capable of self-renewal.

Objective To study the morphologic changes of immunotactoid glomerulopathy and to investigate the clinical pathological features and differential diagnosis.Methods Renal biopsy was observed under the light microscope, immunofluorescence and electron microscopy in a case of newly diagnosed immunotactoid glomerulopathy.Results This patient clinically presented with nephrotic syndrome and hypertension, without family history of renal diseases.Light microscopy showed that diffusely massive and specific protein deposition in the glomerulus in Masson staining.Immunofluorescence revealed IgG, C3 andκwere deposited along the capillary walls and mesangial regions.Electron microscopic examination showed that a large amount of microtubule like substances and a small amount of long bar-shaped and dense crystal-like substances were deposited in the subendothelial spaces and mesangial areas.Conclusions Light microscopy and immunofluorescence of immunotactoid glomerulopathy show no specifically pathological changes.Under electron microscope, a large amount of microtubule like substances is deposited in the glomerulus, which is the key point to distinguish this disease from other glomerular diseases.Except for the microtubule-like substances, the present case is accompanied by the deposition of long bar-shaped and dense crystal-like substance, which has not been reported in previous studies.

Objective To explore the predictive value of serum soluble osteoclast-associated receptor(sOSCAR)for cardiovascular adverse events after percutaneous coronary intervention(PCI)in patients with acute non-ST segment elevation acute myocardial infarction(NSTEMI).Methods From January 2020 to Jan-uary 2022,124 NSTEMI patients admitted to Weifang People's Hospital who underwent PCI were selected as the NSTEMI group,and another 100 healthy volunteers in the same period were selected as the control group.The NSTEMI patients were classified into 31 cases of poor prognosis and 93 cases of good prognosis group ac-cording to whether major adverse cardiovascular events occurred 1 year after PCI.Multivariate Logistic regres-sion model was constructed to analyze the factors affecting the prognosis of NSTEMI patients after PCI,and receiver operating characteristic(ROC)curve was drawn to analyze the predictive value of sOSCAR level on the prognosis of NSTEMI patients after PCI.Results Compared with the control group,serum angiopoietin-2(Ang-2)levels were increased and sOSCAR levels were decreased in the NSTEMI group(P＜0.05).At 1-year follow-up,the incidence of poor prognosis after PCI in 124 NSTEMI patients was 25.00％(31/124).Multivariate Logistic regression analysis showed that increasing age and Ang-2 were independent risk factors affecting the prognosis after PCI in patients with NSTEMI(P＜0.05),and elevated left ventricular ejection fraction and elevated sOSCAR were independent protective factors(P＜0.05).ROC curve analysis showed that the area under the curve of serum Ang-2 and sOSCAR levels combined to predict the prognosis after PCI in NSTEMI patients was 0.865,which was greater than that of serum Ang-2 and sOSCAR levels alone,which were 0.791 and 0.786(P＜0.05).Conclusion Serum sOSCAR level is decreased in patients with NSTEMI and is closely related to the poor prognosis of patients after PCI.Serum sOSCAR combined with Ang-2 has a high predictive value for the poor prognosis of NSTEMI patients after PCI.