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Objective To investigate the effect of CoCl2-induced hypoxia on the expressions of hypoxia-inducible factor-1α(HIF-lα),glucocorticoid receptor(GR)and opiomelanocortin(POMC)in AtT-20 cells.Methods AtT-20 cells were exposed to different concentrations(25,50,100,and 200 μmol/L)of CoCl2 to induce hypoxia.MTT assay was employed to assess the changes in the cell proliferation after the exposure,and real-time PCR and Western blotting were performed to detect the expressions of HIF-1α,GR and POMC genes at both the mRNA and protein levels.Results Compared with the cells in normoxic conditions,the AtT-20 cells exposed to CoCl2 treatment at 25,50,and 100 μmol/L for 24 and 48 h showed obviously enhanced cell proliferation.CoCl2 significantly increased the mRNA and protein expressions of HIF-1α,GR and POMC genes in a dose-dependent manner(P＜0.05).Conclusion CoCl2-induced hypoxia up-regulates the mRNA and protein expressions of HIF-1α,GR and POMC genes in AtT-20 cells.
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Objective To construct the eukaryotic expression vector of mutants in the neurofibromatosis type 2 (NF2) tumor suppressor gene and observe its expression in rat schwannoma cell line RT4. Methods Site-directed mutagenesis was performed to induce the mutation of the codons for the residue Ile 546 in pEGFP-N1-NF2 into Met to construct the muton of pEGFP-N1-NF2~(△Ⅱe546Met). After lipofectin-mediated transient transformation of RT4 with the plasmids containing the mutation and the one without mutation, respectively, the mRNA and protein expression levels of NF2 were determined using fluorescence imaging and Western blotting. The cell proliferation was determined by the MTT assay. Results DNA sequence analysis confirmed the success of site-directed mutagenesis and Western blotting showed that pEGFP-NF2 protein could be expressed in the RT4 cells. The RT4 cell inhibition rate in the pEGFP-N1-NF2~(△Ⅱe546Met) transfection group was statistically lower than that in the pEGFP-N1-NF2 transfected group (P<0.05).Conclusion The recombinant plasmids pEGFP-N1-NF2~(△Ⅱe546Met) has been successfully constructed with efficient expressions in RT4.
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Objective To investigate the expression of free fatty acid receptor GPR40 and evaluate the possible function of GPR40 in the adult monkey hippocampus after ischemia. Methods According to the post-ischemic adult monkey model of Yamashima, a total of 24 adult monkeys were randomly divided into 4 groups: sham-operated group (control) and post-ischemic day 4, 9, 15 (d4, d9,d15). The expression of free fatty acid receptor GPR40 was detected at the protein level by immunoblotting and immunohistochemistry in the adult monkey hippocampus. Results Immunoblotting analysis showed the expression of GPR40 was decreased in CA1 and increased in DG after ischemia (posfischemic group vs control, P＞0.05). Immunohistochemistry data revealed that the double stained cells of GPR40 and NeuN were also decreased by 50% (d15) in CA1 and had no significant changes in DG after ischemia. Interestingly, the co-labeled cells of GPR40 and GFAP were increased 2.5 folds (d4) in post-ischemic SGZ. Conclusions There is the different expression of GPR40 in adult monkey hippocampal CA1 and DG regions after ischemia. Co-labeled cells of GPR40 and GFAP are increased in post-ischemic SGZ, which indicates that polyunsaturated free fatty acid such as DHA, a ligand of GPR40, may alleviate neuronal injury in post-ischemic hippocampus.
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Objective To investigate the effects of exogenous fragile histidine triad(FHIT) gene on apoptosis of human glioma cell line U87. Methods By the method of liposome transfection,plasmids pcDNA3.1/myc-His(-)B-FHIT and pcDNA3.1/myc-His(-)B were transfected into glioma cell line U87.U87 cells were divided into three groups: U87-FHIT group,U87 cells transfected by plasmids pcDNA3.1/myc-His(-)B-FHIT;U87-vector group,U87 cells transfected by plasmids pcDNA3.1/myc-His(-)B;and blank control group,U87 cells without transfection.The expression of exogenous FHIT protein was detected by Western blot and immunofluorescence staining.The effects of FHIT on the growth characteristics of U87 were observed by MTT and flow cytometry. Results Growth inhibitory rate and apoptosis rate of the cells in U87-FHIT group were significantly higher than those in U87-vector group and blank control group(P
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Fragile histidine triad(FHIT) gene is a tumor suppressor gene that locates on chromosome 3p14.2.FHIT can induce cell apoptosis and inhibit cell growth by activating caspase,inhibiting PI3K-Akt-survivin signal pathway and phosphorylation of I?B-?,and binding with microtube.The inactivation of FHIT is closely related with carcinogenesis.The advances in research on the structure,biological function,relationship between inactivation and carcinogenesis,and gene therapy of FHIT are reviewed in this paper.
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Cerebral arteriovenous malformation is a common cerebrovascular disease.Its exact pathogenesis remains unclear.At present,it is thought that this disease is caused by kinds of factors,including congenital and acquired factors.
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<p><b>BACKGROUND</b>Schwannoma is the tumor arising mainly from the cranial and spinal nerves. Bilateral vestibular schwannoma is the hallmark of neurofibromatosis type 2 (NF2). The NF2 gene has been cloned with comprehensive analysis of its mutations in schwannoma. However, most studies focused on vestibular schwannoma. There are differences in proliferation of tumor cell and ultrastructure between vestibular and spinal schwannomas. It is unknown whether genetic alterations in vestibular schwannoma are different from those in non-vestibular schwannoma. We analyzed the loss of heterozygosity (LOH) on chromosome 22 in patients with sporadic schwannoma including vestibular and spinal schwannomas and correlated this genetic alteration with tumor proliferation.</p><p><b>METHODS</b>In 54 unrelated patients without clinical NF1 or NF2, 36 patients had sporadic vestibular schwannoma, and 18 dorsal spinal root schwannoma. Four highly polymorphic linkage to NF2 gene microsatellite DNA markers (D22S264, D22S268, D22S280, CRYB2) were used to analyze LOH. The proliferative index was evaluated by Ki-67 and proliferative cell nuclear antigen (PCNA) immunostaining. Student's t test was used to analyze the difference of the proliferative index between schwannoma with LOH and that without LOH. The difference of the frequency of LOH in vestibular and spinal schwannomas was investigated by the chi-square test.</p><p><b>RESULTS</b>Twenty-three schwannomas (42.6%, 23/54) showed allele loss. The frequency of LOH in vestibular schwannoma was significantly higher than that in spinal schwannoma (chi2 = 5.14, P < 0.05). The proliferative index of schwannoma with LOH was significantly higher than that without LOH (tki-67 = 2.97, P = 0.0045; tPCNA = 2.93, P = 0.0051).</p><p><b>CONCLUSIONS</b>LOH on chromosome 22 is a frequent event in the tumorigenesis of sporadic schwannoma. And, there is a correlation between LOH on chromosome 22 and proliferative activity in schwannoma. The frequency of LOH in vestibular schwannoma is significantly different from that in spinal schwannoma.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Cell Proliferation , Chromosomes, Human, Pair 22 , Genes, Neurofibromatosis 2 , Loss of Heterozygosity , Neurilemmoma , Genetics , Pathology , Neuroma, Acoustic , Genetics , Spinal Cord Neoplasms , Genetics , Spinal Nerve RootsABSTRACT
<p><b>OBJECTIVE</b>To study the clinical features and treatments of jugular foramen neurinomas. The approximate approach and proper exposure regions of jugular foramen tumour were discussed in this paper.</p><p><b>METHODS</b>Fourteen cases of jugular foramen neurinomas were diagnosed by CT, MRI, DSA and 3D-CT reconstruction preoperatively. The tumours were resected by far lateral infra-temporal approach. The classification and relative operative approaches were discussed.</p><p><b>RESULTS</b>Among 14 patients, total removal were achieved in 8 cases, subtotal removal in 4 cases and partial removal in the other 2 cases. All patients got good recovering. No serious complications correlating the approaches occurred except one case of facial paralysis.</p><p><b>CONCLUSION</b>The far later infratentorial approach was suggested to be more suitable for surgery upon neurinomas of jugular foramen.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Angiography, Digital Subtraction , Cranial Nerve Neoplasms , Diagnosis , General Surgery , Craniotomy , Methods , Follow-Up Studies , Imaging, Three-Dimensional , Magnetic Resonance Imaging , Microsurgery , Neurilemmoma , Diagnosis , General Surgery , Tomography, X-Ray ComputedABSTRACT
<p><b>OBJECTIVE</b>To elucidate the role of S-100B and neuron specific enolase (NSE) in predicting the outcomes of patients with severe head injury.</p><p><b>METHODS</b>Forty patients with severe head injury were included in this study. The serum concentrations of S-100B and NSE were measured within 12 hours after head injury to investigate the correlation between serum levels of S-100B and NSE and outcome. Validity of both S-100B and NSE in outcome prediction was assessed with Receiver Operator Characteristic (ROC) curve.</p><p><b>RESULTS</b>The serum concentrations of S-100B and NSE of both groups, with favorable or unfavorable outcomes, were significantly higher than those of the normal group. The serum concentrations within 12 hours after head injury were closely correlated with the prognosis. Furthermore, according to the ROC curves of S-100B and NSE, S-100B was found better in predicting outcomes than NSE.</p><p><b>CONCLUSIONS</b>S-100B and NSE may play important roles in outcome prediction after severe head injury. Moreover, S-100B is clearly superior to NSE in terms of predictive value and appears to be a more promising serum marker in outcome prediction after severe head injury.</p>
Subject(s)
Female , Humans , Male , Craniocerebral Trauma , Blood , Glasgow Outcome Scale , Nerve Growth Factors , Phosphopyruvate Hydratase , Blood , Prognosis , ROC Curve , S100 Calcium Binding Protein beta Subunit , S100 Proteins , Blood , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To estimate outcomes of patients with acute subdural hematomas by analysing the hematoma thickness, midline shift and the differences between them.</p><p><b>METHODS</b>Ninety-five patients with acute subdural hematoma were retrospectively studied by calculating hematoma thickness, midline shift and their difference with a statistical analysis of Kaplan-Meier, Wilcoxon-Mann-Whitney U test.</p><p><b>RESULTS</b>The hematoma thickness ranged from 5.0 to 40.0 mm and midline shift was from 0 to 35.0 mm. Among these patients, 51% died and 49% survived after surgery. 18 patients (19%) showed good or satisfactory results. Kaplan-Meier analysis proved that the survival for patients with hematoma thickness approximately equal to l7 mm and a midline shift 15 mm or whose midline shift exceeded hematoma thickness by 2.2 mm, the survival rate was 50%. Glasgow outcome scale scores were correlated significantly with these parameters.</p><p><b>CONCLUSION</b>The hematoma thickness, midline shift and their difference provided a database from which criteria could be derived, that is crucial for prognosis estimation.</p>