Preliminary trial of identification of brain structures in nude mice with green fluorescent protein expression by fluoroscope / 中华神经医学杂志

Objective To explore the feasibility and advantage of fluoroscope in identification of brain structures in nude mice with green fluorescent protein (GFP) expression. Methods We laid the whole brain separated from 8-week adult nude mice with GFP expression into SLY mouse brain blocker to produce slices of 1 or 0.9 mm thickness; and then,25 μm-thickness frozen sections were cut.Fluoroscope was employed to observe the morphological structure to define their anatomic structures with reference to The Mouse Brain in Stereotaxic Coordinates compiled by Paxinos. After the observation,these frozen sections were performed Nissi staining for contrast. Results Different structures can be identified by their distinct fluorescence intensity:the dense areas of nuclei,Nissl bodies and nerve tract showed low fluorescence intensity; while the structures around the areas of nuclei and nerve tract,such as,the plexiform layer of olfactory bulb and the molecular layer of cerebella,showed high fluorescence intensity.The fluorescence intensity was attenuated obviously after Nissl staining; the visualized structural information observed under stereomicroscope was in accordance with that viewed by fluoroscope.Conclusion The identification of brain structure in nude mice with GFP by fluoroscope can serve as an experimental platform being applied in the anatomic structure positioning in fluorescence tracer experiments.

High-risk factors of progressive hemorrhagic injury after acute traumatic brain injury and its time for first scheduled brain CT / 中华神经医学杂志

Objective To investigate the high-risk factors of progressive hemorrhagic injury (PHI)after acute traumatic brain injury(TBI)and discuss its time for first scheduled brain CT.Methods Retrospective analysis of clinical data of 329 adult patients with blunt TBI,admitted to second affiliated hospital of Suzhou University from August 2009 to July 2010,was performed.Patients were divided into PHI high-risk group and non-PHI high-risk group based on whether clinical symptoms worsened before the first routine scheduled brain CT;and control study was established. Results Forty-one patients from the PHI high-risk group were performed first routine scheduled brain CT ahead of time([3.67±0.96]h)for appearing worsened clinical symptoms,while 288 patients with non-PHI high-risk group were performed first routine brain CT as schedule(8.38±3.03 h);significant difference on the time for the first brain CT was noted between the 2 groups(P<0.05).Statistical differences in aspects of intracranial hematoma volume>10 mL,intracranial hematoma type≥2,associated subarachnoid hemorrhage on initial brain CT,disturbance of consciousness,pupil dilation and scores of Glasgow Coma Scale≤12 on admission were noted between the 2 groups(P<0.05),and statistical differences in aspects of reduced platelet(Plt),prolonged prothrombin time(PT),prolonged activated partial thromboplastin time(APTT),decreased Fibrinogen(FIB)on admission,bilateral brain injury,associated brain contusion,and skull fracture with epidural hematoma on initial brain CT were found between the 2 groups(P<0.05).Logistic regression analysis showed that intracranial hematoma volume>10 mL on initial brain CT, disturbance of consciousness and pupil dilation on admission were predictors for PHI high-risk patients (P<0.05). Conclusion For patients with intracranial hematoma volume>10 mL on initial brain CT, disturbance of consciousness and pupil dilation on admission within 2 h of TBI, first routine scheduled brain CT should be performed within 3-4 h of TBI or even earlier.

Effects of nitric oxide on mitochondrial permeability transition and cytochrome C of human hepatocellular carcinoma cell lines / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the effects of nitric oxide on mitochondrial permeability and cytochrome C (cyt C) of human hepatocellular carcinoma cell lines.</p><p><b>METHODS</b>NO-mediated apoptosis in human hepatocellular carcinoma cell lines SMMC-7721 and HepG2 was investigated by flow cytometry. The growth and proliferation of human hepatocellular carcinoma cell lines SMMC-7721 and HepG2 were evaluted by MTT assay. Mitochondrial transmembrane potential was analyzed by flow cytometry with double staining of Rh123 and PI, and cytoplasmid cyt C was measured by Western blot. The cells were preincubate with cyclosporin A or GSH synthesis blocker BSO to explore their effect on the results of the above experiments.</p><p><b>RESULTS</b>NO donor sodium nitroprusside (SNP) induced apoptosis in human hepatocellular carcinoma cell lines SMMC-7721 and HepG2 and resulted in the decrease of the mitochondrial transmembrane potential and the increase of the amount of cytoplasmid cyt C in time-dependent manner. Cyclosporin A (CsA) specific inhibitor of the mitochondrial permeability transition pore could partially prevent the decrease of delta psi m and the release of cyt C. In contrast, GSH synthesis blocker BSO promoted the decrease of delta psi m and the release of cyt C.</p><p><b>CONCLUSIONS</b>NO may induce apoptosis in human hepatocellular carcinoma cell lines SMMC-7721 and HepG2 by decreasing delta psi m, opening the mitochondrial permeability transition pore, and releasing the cyt C.</p>

Structural analysis of 22 full-length hepatitis B virus genomes isolated from patients with hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the characteristics of full-length Hepatitis B Virus (HBV) genomes isolated from patients with hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The full-length HBV genomes from the serum of HBsAg positive HCC patients were amplified by PCR, and then sequenced and analyzed its structure.</p><p><b>RESULTS</b>Twenty-two full-length HBV DNAs were obtained from different patients of HCC. Phylogenetic analysis revealed that all HBV strains could be categorized into genotype B or C and serotype adr or adw2. Structural analysis showed that HBV obtained shared meaningful consensus mutations in B/T cell epitopes of surface and core antigens, transactivating domain of X protein and enhancer II/core promoter regions as compared to standard strains.</p><p><b>CONCLUSION</b>Genotype and gene mutation of HBV may be closely correlated with the carcinogenesis of HBV-related hepatocellular carcinoma.</p>

Expression and significance of nitric oxide synthase in human hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression and significance of nitric oxide synthase (cNOS) mRNA in primary hepatocellular carcinoma (HCC), cirrhotic liver and normal liver tissue.</p><p><b>METHODS</b>cNOS mRNA expression in 80 HCC, 40 cirrhotic liver and 20 normal liver tissue were observed by in situ hybridization. CD34 immunostain was used to measure the microvascular density (MVD) and Ki-67 immunostain to proliferative index.</p><p><b>RESULTS</b>Expression of cNOS mRNA was observed in the liver cancer cells, endothelial cells in the non-cancerous liver tissues and mononuclear and/or phagocytes. Expression of cNOS mRNA in tumor cells of HCC was higher than that in the liver cells of cirrhotic liver (P < 0.01) which was higher than the normal liver tissue. Expression in the endothelial cells was higher in HCC and cirrhotic liver than those in the normal liver tissue (P < 0.01). HCC with positive cNOS mRNA expression in the endothelial cells showed higher extent of neovascularization and degree of proliferative index. The more MVD, the higher proliferative index, which increased in metastatic tumors.</p><p><b>CONCLUSION</b>cNOS mRNA expression was involved in oncogenesis, angiogenesis and progression of hepatocellular carcinoma.</p>