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BACKGROUND: Diabetes can lead to many complications. Of them, retinopathy is a common type. To explore the effect of erythropoietin in diabetic retinopathy, it is necessary to establish an animal model of similar pathologic features and high reproducibility in clinical retinal neovascularization. OBJECTIVE: To investigate the protective effect of vitreous cavity injection of erythropoietin on optic nerve of rats with diabetic retinopathy. METHODS: Rat models of diabetic retinopathy were established and injected with erythropoietin through vitreous space for 14 consecutive days. Results were compared between the model and normal control groups. RESULTS AND CONCLUSION: (1) Transmission electron microscope: compared with the model group, retinal ganglion cel nucleus presented uniform red; cel membrane was intact; endoplasmic reticulum exhibited slight swel ing. A smal number of vacuoles were found in mitochondria around the nuclei. (2) Western blot assay and RT-PCR: compared with the model group, Caspase-3 and Bax protein expression was lower in the erythropoietin group (P < 0.05). Caspase-3 protein and Bcl-2 protein expression was significantly higher in the erythropoietin group than in the model group (P < 0.05). Significant differences in nuclear factor κB and Survivn gene expression were detected between the model and erythropoietin groups (P < 0.05). (3) Results verified that vitreous cavity injection of erythropoietin had neuroprotective effect on rats with diabetic retinopathy, which may be associated with the inhibitory effect on apoptosis.
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Objective To investigate the clinical application and diagnostic value of impulse oscillometry lung function in children with bronchial asthma.Methods Thirty-two children (19 children in acute attack group,13 children in clinical remission group) with bronchial asthma were selected,and 20 healthy children as control group.The lung function simation by Master Screen impulse oscillometry device was detected.Results The outcome of the R5,R20,R5-R20,the negative of X5 and Fres in acute attack group (21.90±2.70) Hz]were higher than those in clinical remission group [(7.75±0.97),(3.82±0.50),outcome of the R5,R5-R20 and Fres in clinical remission group were higher than those in control group (P＜0.05),but there were no statistical significance in R20 and X5 in the two groups (P＞0.05).Conclusion Impulse oscillometry can provide various objective indicators to bronchial asthma in acute attack and clinical remission stage.which can be used as a method in detecting pulmonary function and monitoring therapy of bronchial asthmatic children.
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BACKGROUND: Compared with bone marrow-derived mesenchymal stem cells (MSCs),human umbilical cord blood (hUCB)-MSCs possess many advantages,including easy acquirement,low immunogenicity,able to tolerance a higher degree of HLA-matching inconsistency,and with higher purity.OBJECTIVE: To analyze the method and condition for in vitro isolation,purification,proliferation,and osteoblast and lipoblast differentiation of hUCB-MSCs.DESIGN,TIME AND SETTING: The present observational experiment was performed in the Key National Laboratory of Biological Treatment,Huaxi Hospital,Sichuan University (i.e.,Institute of Stem Cells and Tissue EngIneering) between September 2004 and November 2005.MATERIALS: Neonatal cord blood at gestational age 37 to 40 weeks.METHODS: hUCB-MSCs were collected in aseptic condition,isolated by density gradient centrifugation,sedimenting red cells with methylcellulose followed by density gradient centrifugation,or immunomagnetic beads negative technique of CD34+.Theisolated MSCs were used to carry on plastic adherent culture in L-DMEM +10% fetal bovine serum (FBS) or MesencultTM medium +10% FBS.The third passage of cells were used for surface antigen determination by flow cytometry and induced to differentiate towards osteoblasts and lipoblasts.MAIN OUTCOME MEASURES: ①Identification of hUCB-MSCs.②Confirmation of hUCB-MSC differentiation by alizarin red and oil red staining.RESULTS: The mononuclear cells isolated by sedimented and centrifuged way cultured in MesencultTM medium +10% FBS were most available.Obvious colonies appeared in the third passage.The cells obtained by only centrifugation in density gradient were hard to form colony,and those isolated by immunomagnetic beads were hard to culture.The surface antigens of these colony cells presented CD29,CD59,and CD71,hut did not express CD34,CD45,and HLA-DR,and so on.After alizarin red staining,osteoblast-differentiating colony cell cytoplasm exhibited mineralized matrices.After old red staining,lipoblast-differentiating colony cell cytoplasm was full of lipid vacuoles.CONCLUSION: The MSCs can be successfully isolated by sedimenting red cells with methylcellulose followed by centrifugation and cultured in MesencultTM medium +10% FBS.Obvious colony growth appears in the third passage.After induction,the MSCs can differentiate into osteoblasts and lipoblasts.
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BACKGROUND: Diabetic retinopathy is a commonly chronic-vascular complication in a course of diabetes mellitus (DM), and its relation with apoptosis and changes of ion level needs to be proved.OBJECTIVE: To observe apoptosis and changes of Na+ and K+ contents in retinal tissue of DM rats.DESIGN: Randomized controlled study.SETTING: Department of Ophthalmology, Liuzhou Municipal People's Hospital.MATERIALS: The experiment was completed at the Department of Ophthalmology, Liuzhou Municipal People's Hospital. A total of forty adult male Wistar rats, aged 20 months, weighing 320-350 g, were provided by Animal Center of Medical College Affiliated to Sun Yat-sen University.METHODS: Forth Wistar rats were randomly divided into control group and experimental group with 20 in each group. Streptozotocin was used to induce DM in the experimental group. Apoptosis and changes of Na+ and K+ contents in retinal tissue were measured on the 4th, 6th, 12th and 16th weeks after DM onset. Rats in the control group were injected only with the same volume of citrate buffer solution. Then, correlations on the aspect of fasting blood glucose (FBG), hemoglobin glycosylation (HbAlc), Na+ and K+ contents and DM course were analyzed between the two groups.MAIN OUTCOME MEASURES: ① Na+ and K+ contents; ② FBG concentra tion and HbAlc level; ③ changes of apoptosis; ④ correlations among markers.RESULTS: Apoptosis could be detected in retinal tissue in the experimental group at 4 weeks after DM onset, and with course elongating, level of apoptosis was aggravated gradually. Na+ concentration was increased in retinal tissue, but K+ concentration was decreased (P ＜ 0.01 or 0.05). Levels of apoptosis in retinal tissue in the experimental group were positive correlation with FBG concentration, HbAlc level, Na+ content and DM course (r=7.584, 7.844, 7.369, 6.246; P ＜ 0.01); however, they were negative correlation with K+ content in retinal tissue (r=7.658, P ＜ 0.01).CONCLUSION: There are apoptosis and abnormal Na+ and K+ contents in retinal tissue of DM rats. Moreover, these changes may be one of pathological bases of diabetic retinopathy.