AMP-Activated Protein Kinases ; genetics ; Heart Diseases ; Humans ; Syndrome

Background Establising the culture model of Müller cells for obtaining the highly putified target cells is essential for the study about the physiology and pathology of retinal Müller cells. The exsiting purifing method for culturing Müller cells is dissatisfactory. Objective This study was to establish a method to obtain high purifing Müller cells. Methods The retina from 5 clean newborn SD rats were isolated and digested by 0. 01% trypsin and cultured in DMEM containing 10% fetal bovine serum. The cellular suspension was then prepared,and the target cells were screened using flow cytometry based on the size and the quantity of cells. Cultured and passaged cells were identified by transmission electron microscope and light microscope. Immunocytochemistry was used to detecte the expression of GFAP in cultured cells for the determination of type and purity of the cells. Results The cells showed the similar shape to retinal Müller cells after primarily culture with the large volume, and some small other types of cells could been seen. The growth of cells was quickly 3 weeks later. The fibroblasts were removed using sticking-wall by steps,and neurons were eliminated following passage. Aboundent of cellular organs were seen under the transmission electron microscope. The positive response rate of the cells for CFAP was 100%. Conclution Flow cytometry offer a rapid and feasible approach for purifying Muller cell and it builds the foundation for further study about Müller cells.

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BACKGROUND: In the central nervous system(CNS) of normal adults there are neural stem cells or neural progenitor cells, which are capable of self-renewing and multiple differentiating. In normal physiological conditions, intrinsic neural stem cells are in a resting state. What state will they be in during cerebral ischemia?OBJECTIVE: To observe the distribution, proliferation and differentiation of intrinsic neural stem cells in focal transient ischemia in rats.DESIGN: A randomized controlled exploratory trial based on the rats.SETTING: Neurobiological department, histological and embryological department of a medical college.MATERIALS: The experiment was conducted in the Neurobiological Department of Luzhou Medical College from July 2001 to July 2002. Altogether 41 healthy adult SD rats of either gender, weighting 250 g - 300 g, were selected.INTERVENTIONS: The focal ischemia model was made by blocking middle cerebral artery(MCA) and reperfusing for 0.5 hour, 3 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 5 days and 10 days. Sham-operation group was treated by the same method, but the filament was not long enough to block MCA, and normal rats served as control group. The rats were sacrificed at given time points, and their brains were made into cerebral slices. The single-and double-labeled immunohistochemical staining was employed to detect the proliferation, distribution and differentiation of intrinsic neural stem cells.MAIN OUTCOME MEASURES: The distribution, proliferation and differentiation of intrinsic neural stem cells.RESULTS: Immunoreactivity of proliferating cell nuclear antigen(PCNA)was present in most ependymal cells in ventricular zone(VZ), and PCNA-positive cells were sparsely distributed in the parenchyma in normal and sham-operation groups. At 3 hours of reperfusion, PCNA-labeled cells were first detected in rostral subventricular zone. At 12 hours of reperfusion and onward, PCNA-positive cells appeared in some choroid plexus cells in bilateral lateral VZ. At day 3 to day 10 of reperfusion, PCNA-labeled cells significantly increased in infarct boundary in preoptic area, striatum and deep layer of frontoparietal cortex. PCNA-labeled cells were first detected in subgranular zone of dentate gyrus 3 days after reperfusion, and increased with time. A very small number of double-positive cells expressed with PCNA and glial fibrillary acidic protein(GFAP) were first detected in infract boundary in preoptic area on day 3 and onward. No double-PCNA and NF-positive cells were detected within 10 days of reperfusion.CONCLUSION: Focal cerebral ischemia activates intrinsic neural stem cells, which proliferate and differentiate, and migrate toward ischemic striatum and frontoparietal cortex. This may help clarify the mechanism of functional recovery after ischemia.

Aim To investigate glutamate-induced [ Ca2 + ] i changes in cultured rat neonatal cortical astrocytes after hypoxia/reoxygenation, H2O2 or high concentration of L-glutamate injury. In the meantime, the effects of Gingko biloba extract (GbE) were examined. Methods [ Ca2+ ]i changes in astrocytes were monitored by laser scanning confocal microscopy with the Ca2+ sensitive fluorescent probe [ Ca2 + ] i, but decrease by ( 3.3 ± 1.6) %, (81 ± 11 ) % and ( 81 ± 7 ) %, respectively. Pretreatment with H2O2 or high concentration of L-glutamate injury were ( 135 ± 98 ) %, ( 117 ± 93 ) % and ( 89 ± 36) %,different injury. Conclusion Hypoxia/reoxygenation, H2O2 and high concentration of L-glutamate impaired astrocytes' response to exogenous L-glutamate, and then bidirectional communication between astrocytes and neurons could not take place. GbE could improve the abnormal responses and maintain the normal function of astroglical network. These effects support that GbE has potential beneficial actions against brain injury.

Objective To detect the expression of Ezrin in giant-cell tumor of bone,and to investigate its cilincal significance. Methods 60 cases of biopsy which had been confirmed as bone giant-cell tumors in our hospital from January 2008 to December 2013 were set as observation group;tumor tissues from 8 cases of reactive new bone in nonmalignant bone diseases,12 cases of osteoid osteoma and 11 cases of osteoblastoma in the corresponding period were set as control group. Protein and gene levels of Ezrin were tested with Western blotting method and real-time PCR detection,simultaneously proceeded the corresponding analysis combined with the clinical data of patients;60 cases of bone giant-cell tumor patients accepted tumor resection and pros-thesis replacement,2 courses of preoperative chemotherapy;mitochondria morphological changes of tumor tissue and Ezrin protein and genetic changes were observed before and after chemotherapy. Results In the giant-cell tumors of bone,the Ezrin protein mainly located in the cytoplasm,and its expression positive rate was much higher than that in reactive new bone of nonmalignant bone diseases(19. 7% ),osteoid osteoma(21. 2% )and osteoblastoma(20. 7% );the difference was statistically significant(χ2 = 4. 18,P = 0. 024),but no statistical difference in the Ezrin expression among the groups of osteosarcoma,osteoid osteoma and osteblastoma(χ2 =6. 18,P = 0. 087). In the giant-cell tumors of bone tissue after chemotherapy,mitochondria pyknosis and the phenomenon of liquid cavitation was less than that before the treatment,and Ezrin protein expression decreased and gene levels reduced[(23. 99 ± 1. 49)vs(20. 11 ± 1. 11),t = 5. 03,P = 0. 018)]. Conclusion The expression of Ezrin in giant-cell tumor of bone is much higher than other benign bone tumor,and it could be a biological marker for differentiating benign and malignant bone tumor. Early intervention in Ezrin may be helpful for reatment of giant-cell tumor of bone.

Objective To investigate the relationship of epidermal growth factor receptor (EGFR) mutation with clinical features of baselines as well as serum CEA level in patients with recurrent non-small cell lung cancer (NSCLC). Methods A total of 54 patients with first recurrence of advanced lung cancer who had received chemotherapy were included in this study. ADx-ARMS was performed to detect EGFR gene mutations in surgical specimens taken from the primary tumor. Serum CEA level was measured by the electrochemical luminescence method. Results The mutation rate of EGFR was significantly higher in females than in males (χ2= 11.868, P =0.006), with a total mutation rate of 60.8%in 106 patients. The rate was higher in adenocarcinoma than in other histological types(χ2=6.002,P=0.014), and significantly higher in non-smokers than in smokers (χ2= 8.502,P=0.004) and in the patients with serum CEA level over or equal to 5.0 ng/mL than those with CEA level less than 5.0 ng/mL (χ2=22.543,P=0.000). A multivariate analysis revealed that a higher serum CEA level at the time of disease recurrence was associated with EGFR gene mutations (P = 0.002). Conculsions Serum CEA level is closely associated with the presence of EGFR gene mutations in patients with first recurrence of advanced NSCLC. A higher serum CEA level at the time of disease recurrence is independently associated with EGFR gene mutations. CEA level can be used as a potential indicator to determine EGFR mutation.

Objective To investigate the diagnostic value of determination of the genotypes in codon 12 and 13 of K-ras oncogene in blood samples of patients with pancreatic carcinoma(PC).Methods Blood samples were obtained from 54 patients with pathologically confirmed PC,and 33 healthy controls.The DNAs were obtained in these samples.and then genotype of K-ras mutation was detected by using the PNA-clamping real-time quantitative PCR.Then the correlation between the K-ras genotypes of blood DNA and the clinical characteristics was analyzed.Results K-ras mutations were found in 74.1%(40/54)of patients with PC.There was no such mutation in control samples.The mutations of K-ras was associated with age,lymph node and vessel invasion.poorly differentiated tumor,CA19-9,while it was not associated with sex,tumor location,size of tumor,clinical staging and pathological type.Conclusions The one-step method was highly sensitive for detecting K-ras mutation in blood samples.Detection of circular blood cells harboring K-ras mutation suggested the tumor was highly invasive with poor prognosis.

lso.This result suggested that Th17 subset is differentiated in chronic stage of viral myocarditis.