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Objective To construct a scientific behavioral research of pasta matrix reaching task (PMRT) and comprehensively evaluate sensory-motor dysfunction caused by brain injury.Methods Twenty-one SD rats were subjected to 14-days pasta matrix grasping training and then were randomly divided into model group (11 rats) and sham group (6 rats).Motor cortex ischemia was induced by injection of endothelin-1 in SD rats.The number of pasta grabed by the injured forelimb and the location in matrix were evaluated daily 7 days after surgery.The infarct volume was measured by Nissl staining at the 7 days,14 days,and 28 days after stroke.Results The number of pasta obtained by rats was reduced from (33.43± 1.02) to (20.57±0.57) at 7 days post stroke in model group,and then increased to (26.85±0.98) at 28 days post stroke,although there was a significant difference between sham group (32.33± 1.45) and ischemic group (t=3.198,P＜0.05).The frequency of retrieval from each slot of the pasta matrix represented that sham group demonstrated a significant gain in performance in the antero quadrant of the matrix compared to ischemic rats by the fourth week after stroke.The stroke volume was decreased from (37.82± 1.17)mm3 at 7 days post-stroke to (24.35±0.38)mm3 at 28 days post-stroke,indicating brain recovery from ischemic injury.Conclusion The pasta matrix reaching task can function as a versatile and sensitive behavioral assay that permits experimenters to collect accurate outcome data and manipulate limb use to mimic human clinical phenomena including compensatory strategies and focused rehabilitative training after stroke.
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Objective@#To construct a scientific behavioral research of pasta matrix reaching task (PMRT) and comprehensively evaluate sensory-motor dysfunction caused by brain injury.@*Methods@#Twenty-one SD rats were subjected to 14-days pasta matrix grasping training and then were randomly divided into model group (11 rats) and sham group (6 rats). Motor cortex ischemia was induced by injection of endothelin-1 in SD rats. The number of pasta grabed by the injured forelimb and the location in matrix were evaluated daily 7 days after surgery. The infarct volume was measured by Nissl staining at the 7 days, 14 days, and 28 days after stroke.@*Results@#The number of pasta obtained by rats was reduced from (33.43±1.02) to (20.57±0.57) at 7 days post stroke in model group, and then increased to (26.85±0.98) at 28 days post stroke, although there was a significant difference between sham group(32.33±1.45) and ischemic group (t=3.198, P<0.05). The frequency of retrieval from each slot of the pasta matrix represented that sham group demonstrated a significant gain in performance in the antero quadrant of the matrix compared to ischemic rats by the fourth week after stroke.The stroke volume was decreased from (37.82±1.17)mm3 at 7 days post-stroke to (24.35±0.38)mm3 at 28 days post-stroke, indicating brain recovery from ischemic injury.@*Conclusion@#The pasta matrix reaching task can function as a versatile and sensitive behavioral assay that permits experimenters to collect accurate outcome data and manipulate limb use to mimic human clinical phenomena including compensatory strategies and focused rehabilitative training after stroke.
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Angiogenesis is a process that new blood vessels generate on the basis of an original vascular plexus via sprouting or other forms during normal growth and development of an organism or under particular conditions such as wound repairing and ischemic hypoxia. In general, angiogenesis includes five stages: degradation of the vascular basement membrane, endothelial cell proliferation, vascular sprouting, lumen formation and vascular network turning stable and mature. Among ischemic heart diseases, acute myocardial infarction seriously threatens human health, its pathological characteristics include myocardial vascular stenosis and blockage,leading to myocardial infarction. Therefore,the promotion of angiogenesis has become one of the therapies for ischemic heart diseases. A variety of pro-angiogenic cytokines are involved in the process of angiogenesis, such as polypeptide growth factors and lipid mediators. In this paper we review the recent progress in research on the expression of pro-angiogenic factors in myocardium after ischemic myocardial infarction.
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Objective To compare the diagnostic ability of gadolinium diethylene-triamine-pentaacetic acid-enhanced magnetic resonance imaging (Gd-EOB-DTPA MRI) and contrast-enhanced ultrasound (CEUS) for the diagnosis of hepatocellular carcinoma (HCC) ≤ 3 cm in diameter in patients with hepatic cirrhosis or chronic hepatitis B.Methods Ninty-two cases with single focal liver lesion ≤ 3 cm who underwent CEUS and Gd-EOB-DTPA MRI in Southwest Hospital Affiliated to Third Military Medical University were enrolled in the study.And all cases were diagnosed with hepatic cirrhosis or chronic hepatitis B.Pathology was the golden standard and all cases were diagnosed pathologically by surgical operation.Sensitivity,specificity,positive predictive value,negative predictive value and accuracy rate of the diagnosis of small HCC by CEUS,Gd-EOB-DTPA MRI and CEUS combined with Gd-EOB-DTPA MRI were calculated and compared with x2 test or Fisher exact probability.Results Of the 92 cases,82 cases were diagnosed as HCC and the other 10 cases were diagnosed as non-HCC.The sensitivity and the specificity for the diagnosis of HCC was 79.2％ (65/82) and 70.0％ (7/10) of CEUS alone and was comparable with that of Gd-EOB-DTPA MRI [74.4％ (61/82) vs 70.0％ (7/10)] alone (x2=0.548,P=0.459;P=1.000).When both CEUS and Gd-EOB-DTPA MRI met the diagnostic criteria for HCC,the sensitivity was lower [61.0％ (60/82),P=0.010] than that of CEUS alone,and the specificity [80.0％ (8/10),P=1.000] was not statistically different from that of CEUS alone.When CEUS or Gd-EOB-DTPA MRI met the diagnostic criteria for HCC,the sensitivity [92.7％ (76/82)] was higher than that of CEUS alone (x2=6.119,P=0.013) or Gd-EOB-DTPA MRI alone (x2=9.972,P=0.002),and the specificity [60.0％ (6/10)] was not statistically different from that of CEUS alone (P=1.000) or by Gd-EOB-DTPA MRI alone (P=1.000).Conclusions In patients with cirrhosis or chronic hepatitis B,the diagnostic ability of HCC ≤ 3cm in diameter by CEUS alone was comparable to that by Gd-EOB-DTPA MRI alone.Combination of CEUS and Gd-EOB-DTPA MRI may increase the sensitivity for the diagnosis of HCC,but the specificity not statistically improved.
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Objective To explore the effects of morroniside on the expression of CD34 in ipsilateral cortex of rats after focal cerebral isch-emia-reperfusion. Methods 45 male Sprague-Dawley rats were divided into sham group (n=9), ischemia group (n=9), and morroniside groups (low, medium and high dosage groups, n=9). The middle cerebral artery were occluded for 30 minutes, and reperfused. Morroniside was administered intragastrically once a day at dose of 30 mg/kg, 90 mg/kg, 270 mg/kg after operation. The expression of CD34 in the isch-emic ipsilateral cortex were detected with immunohistochemistry (n=6) and Western blotting (n=3) 7 days after operation. Results The ex-pression of CD34 increased in the ischemia group compared with the sham group, and further increased in the morroniside groups of high dos-age compared with the ischemia group (F>14.865, P<0.001). Conclusion Morroniside could increase the expression of CD34 in the ischemic ipsilateral cortex after ischemia-reperfusion in rats, which may promote the angiogenesis and neurogenesis after ischemia.
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The establishment of professional master degree in pathology is a new branch of the professional degree graduate education.It is an important measure to bring the clinical pathology into the teaching system of clinical medicine specialty in China.In response to a wave of education reform in the new century, our department has positively converted the traditional postgraduate training and teaching mode, innovated the teaching idea and content, and continued to improve the teaching method and means.On the basis of fully respecting students' subject consciousness, we have combined traditional and modern teaching means, made full use of modern education information technology, actively built and promoted pathology specialized degree graduate student research, teaching and clinical trinity management: the new training mode.Through the practice of bilingual, network, clinical problems and cases of teaching and combined with the leading edge forum of famous pathologists' research and the professional standardized training of clinical pathology, we have extensively carried out the education training, which is based on the theory, practice, research, and sub specialist guidance.
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@#Objective To investigate the effects of morroniside on the expression of vascular endothelial growth factor (VEGF) and fi-broblast growth factor-2 (FGF-2) in rat cortex after focal cerebral ischemia-reperfusion. Methods 30 male Sprague-Dawley rats were ran-domly divided into sham group, model group, morroniside-low group (30 mg/kg), morroniside-middle group (90 mg/kg) and morroni-side-high group (270 mg/kg). Middle cerebral arteries of rats were occluded for 30 minutes with Longa's method and re-perfused. The ex-pression of VEGF and FGF-2 in the ischemic ipsilateral cortex was detected with Western blotting 7 days after reperfusion. Results The ex-pression of both VEGF and FGF-2 increased in the ischemic ipsilateral cortexin in all the ischemic groups compared with the sham group (P<0.05). The expression of VEGF further increased in a dose-dependent manner in all the morroniside groups compared with that of model group (P<0.05), and the expression of FGF-2 increased in the morroniside-high group (P<0.001). Conclusion Morroniside could increase the expression of VEGF and FGF-2 after ischemia-reperfusion, which might promote angiogenesis.
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Objective The purpose of this study is to select suitable ages of rats for the CPR ( cardiopulmonary resuscitation) animal model.The neurological function score and subgroups analysis are evaluated in 2 month old and 4 month old animal groups.Methods Based on the evaluation of physiological indexes including ECG, blood pressure and neurological function defect score ( NDS) and subgroup analysis, the stability of CPR rats model was compared between 2 month old and 4 month old animal groups.Results The results showed that, the model rate of the ventricular fibrillation was induced by electrical stimulation , the 4 month old group was 87.5%, significantly higher than the 2 month old group, however, there was no significant difference between the two groups in the mortality rate;For the changes of blood pressure during the process of CA( cardiac arrest) induced by electrical stimulation, the 4 month old group was significantly lower than the 2 month old group (P <0.01); for the NDS at each time point after CPR, there was no significant difference between the two groups; however, the NDS subgroup analysis at different time points showed that there were different degrees of differences between the two age groups ( P <0.05) .Comparing with the 2 month old group, the 4 month old group had a stable process during the animal model preparation, had an obvious cerebral blood hypoperfusion phenomenon and aggravation of brain injury after CPR.Conclusion The 4 month old rats are more suitable for preparation of CPR animal mode , the model rate is high, the brain injury aggravate.It is more suitable evaluation for basic research and treatment of CPR.
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BACKGROUND: Occurrence and development of the glioma are not only related to regulative gene of cell cycle, but also dealt with adjustive genes of cell invasion, cell metastasis and apoptosis.OBJECTIVE: To identify the differentially expressed genes related to cell proliferation and apoptosis in glioma cell of different differentiating degree with cDNA microarray, and provide basic data for further research of mechanisms of cell proliferation and apoptosis in human gliomas.DESIGN: Opening-up experiment.SETTING: Institute of Pathology, Southwest Hospital and Department of Neurobiology, Basic Medicine Faculty as well as Department of Pharmacy,Xinqiao Hospital, Third Military Medical University of Chinese PLA.MATERIALS: Human malignant glioma cell line CHG-5 ( Ⅱ grading according to the WHO standard) was constructed, kept and cultured in this experiment. SHG-44 (Ⅳ grade according to the WHO standard) was provided by Institute of Brain Tumor, Second Hospital Affiliated to Suzhou Medical College. Serum of calf was produced and provided by Hangzhou Sijiqing Biomaterial Institute. In experiment RPMI-1640 medium (Gibco),Trizol test kit (Gibco-BRL), RNAsecureTM solutions (Ambion, Austin,Texas), biophotometer (Eppendorf, Hamburg, Germany) were also applied.Gene chip contained 9 984 human cDNA segment, which was prepared and provided by City University of Hongkong (UniGEMV2 cloneset known gene and ESTs were purchased from Incyte Company), Superscript Ⅱ reverse transcriptase was provided by Gibco-BRL Company. Fluorochrome Cy3 & Cy5 was the products of Amersham Pharmacia Company.METHODS: The experiment was performed at Chongqing Third Military Medical University of Chinese PLA and City University of Hongkong between 2001 and 2003. Total RNA was extracted from Trizol test kit. Reverse transcription polymerase chain reaction (RT-PCR) was conducted in Superscript Ⅱ reverse transcriptase, and cDNA product was marked with fluorochrome Cy3 & CyS. Followed by chip hybridizationto detect the difference of gene expressions between human glioma cell line CHG-5 and SHG-44 tumor cell, especially the difference of related genic expression between cell proliferation and apoptosis, and the chip result was verified with Northern blot hybridization.MAIN OUTCOME MEASURES: The difference of gene expression of human glioma cell in different differentiating degree; Comparison of result between Northern blot and related gene chip.RESULTS: ①Compared with CHG-5, 120 gene expressions were detected obvious up-regulation and 22 gene expressions were significant down regulation in SHG-44 cells, and the variety of these differentially expressed genes was many, in which apoptosis related genes were 6, including 3 up-regulation genes and 3 down-regulation genes; Cell cycle and proliferating related genes were 12, including 5 up-regulation genes and 7 down-regulation genes. ②Chip result was supported by Northern blot result further.CONCLUSION: The differentially expressed genes of glioma are revealed primarily, especially the differentially expressed genes related with cell proliferation and apoptosis.
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Objective To observe the difference of morphology and phagocytosis between alveolar macrophages (AMs) and pulmonary interstitial macrophages (IMs). Methods AMs were collected by lung lavage and IMs by treatment of the lung tissue with DNAse and collagenase. The two cell populations were analyzed with respect to morphology by transmission electron microscopy, and the variation of these macrophages of phagocytosis were tested by malachite green colorimetry. Results There were great differences in morphology between AMs and IMs. The phagocytosis of AMs was much stronger than that of IMs. Conclusion There is functional and morphological heterogeneity between AMs and IMs. IMs should not be regarded as the precursors to AMs.
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Objective To investigate the expression of survivin and its correlation with tumor cell proliferation and axillary lymph node metastasis in breast cancer. Methods Expressions of survivin and proliferating cell nuclear antigen(PCNA) in 74 cases of breast cancer(34 cases with axillary lymph node metastasis) were measured by immunohistochemistry. Results (1) There was expression of survivin in 64.9% of breast cancer. (2) Labeling idex(LI) of PCNA in breast cancer cells with positive expression of survivin was significantly higher than that with negative survivin. (3) The positive percent of survivin expression in breast cancer with axillary lymph node metastasis was higher than that without axillary lymph node metastasis. Conclusion The expression of survivin in breast cancer is related to PCNA and axillary lymph node metastasis.
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Objective To compare several histopathological staining methods in demonstrating Cryptococcus neoforman.Methods Hematoxylin-eosin(HE) method,periodic acid-Schiff(PAS) method,Alcian blue method and Grocott-Gomori's methenamine silver method were carried out in the lung and brain tissue samples from 7 autoptical cases identified with Cryptococcus neoforman infection.Results All these methods demonstrated Cryptococcus neoforman infection,HE method showed unclear pink cell body which were difficult to be found.Alcian blue method displayed blue picture,and PAS method showed violet-red body,and Grocott-Gomori's methenamine silver method presented clear dark black picture which was easy to be distinguished from other components.Conclusion Grocott-Gomori's methenamine silver method is the best method in staining Cryptococcus neoforman.
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Objective To investigate roles of cyclin-dependent kinase 4 (CDK4) in nordihydroguaiaretic acid (NDGA)-induced inhibitory effect on proliferation of human malignant glioma cells. Methods The techniques of cell culture, cell counts, flow cytometry, immunoprecipitation, immunohistochemistry and image analysis were employed in this study. Results ①A concentration-dependent inhibition of proliferation was demonstrated in the SHG-44 cells incubated for 24 hours in the presence of NDGA, and cell proliferation was blocked in the G1→S phase. ②The activity of CDK4 was decreased apparently in the SHG-44 cells treated for 24 hours with 10 to 200 μmol/L NDGA in a concentration-dependent way. ③The expression of CDK4 gene was downregulated in the cells after NDGA treatment. Conclusion CDK4 plays an important role in NDGA-induced inhibition of glioma cell proliferation.
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Objective To investigate the changes and their significance of bcl-2 and c-myc in nordihydroguaiaretic acid (NDGA)-induced apoptosis of human malignant glioma cell line SHG-44. Methods The apoptosis of SHG-44 cells was observed with light and electron microscopy and TUNEL method. The expression of bcl-2 and c-myc gene was measured with immunohistochemistry, in situ hybridization and image analysis. Results ① The SHG-44 cell apoptosis was induced by NDGA at a concentration lower than 200 μmol/L in a time-dependent manner. ② The expressions of bcl-2 and c-myc gene in SHG-44 cells were decreased after the treatment of 100 μmol/L NDGA with the elapse of time, indicating a close association with cell apoptosis. ③ The expressions of bcl-2 and c-myc mRNA in SHG-44 cells were decreased after the treatment with 100μmol/L NDGA, which was apparently consistent with the immunohistochemical results. Conclusion The NDGA-induced apoptosis in human malignant glioma cells might be related with the down-regulated expressions of bcl-2 and c-myc gene. The exact mechanism needs further research.
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Objective To explore the effect of nordihydroguaiaretic acid (NDGA) on the expressions of vascular endothelial growth factor (VEGF) and its receptor, kinase-inserted domain containing receptor(KDR) and the possible mechanism. Methods The expression of VEGF in human malignant glioma cell line SHG-44 and that of KDR in human umbilical vein endothelial cell (HUVEC) line ECV-304 were observed 1～3 d after NDGA treatment with immunohistochemistry, in situ hybridization and image analysis. Results The expression of VEGF was declined at protein or mRNA levels in SHG-44 cells after treated with 100 μmol/L NDGA for 1 to 3 d. The expression of KDR in endothelial cells with 100 μmol/L NDGA treatment for 1 to 3 d was decreased too, in a more obvious way compared with the decline of VEGF expression in SHG-44 cells. Conclusion The results suggest that NDGA inhibits the expression of VEGF in glioma cells as well as that of VEGF receptor KDR in endothelial cells, which may be the important molecular mechanism of anti-angiogenesis of NDGA.
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AIM: To observe the effect of continuous inhaled nitric oxide (NO) on airway resistance in guinea pigs. METHODS: 36 healthy male guinea pigs were divided into control group, isoproterenol group and two inhaled NO (20?10 -6 and 60?10 -6 ) groups. Respiratory resistance (R_E) and dynamic compliance(C_ dyn )were recorded before and after evoked by histamine at different doses. RESULTS: After injections of intravenous histamine at 80,120 and 160 ?g/kg, the R_E of inhaled NO groups were apparently lower than that of control group. Compared with control group, the C_ dyn of inhaled NO groups were significantly higher after administration of histamine at 80, 120 and 160 ?g/kg. After given histamine at more than 80 ?g/kg,the R_E of inhaled NO groups were higher and the C_ dyn lower than those of isoproterenol group. CONCLUSION: Inhalation of 20?10 -6 NO and 60?10 -6 NO can inhibit the increase in airway resistance induced by higher doses (80,120 and 160 ?g/kg )of intravenous histamine, but the effect of intravenous isoproterenol seems stronger.
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Objective To construct antisense survivin eukaryotic expression vector pIRES2-EGFP-SVVas and detect its expression in human malignant glioma cell line SHG-44 cells. Methods The antisense survivin eukaryotic expression vector pIRES2-EGFP-SVVas was constructed and then transfected into SHG-44 cells with lipofectamine. The expression of survivin on the cells was measured by RT-PCR and Western blotting. Results The mRNA and protein expression of survivin decreased in the transfected cells. Conclusion pIRES2-EGFP-SVVas can decrease the expression of survivin in the cells transfected with the vector.