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Objective To observe the apoptosis of retina and the expression of caspase-3 in mice with premature myopia and to explore the pathogenesis of premature myopia.Methods Together 60 newborn C57BL/6J mice were selected and divided randomly into three groups (n =20):P6 group (opening the eyelid on day 6 after birth),P10 group (opening the eyelid on day 10 after birth) and normal group (opening the eyelid naturally).The right eyes of mice in the P6 and P10 group were subjected to lighting exposure,and the left eyes were left untreated serving controls with its right eyes;while the eyes in the normal group open naturally without any treatment.Then the refraction was checked on day 15 through retinophotoscopy,and ocular axial length was measured by micrometer with electronic digital display.TUNEL assay was used to determine retinal apoptosis.Immunohistochemistry and Western blot were used to detect the expression of caspase-3 in mice retina.Results The right eyes developed significant myopia in the P6 group [(-7.55 ±0.15)D] and P10 group [(-5.25 ±0.10)D],while the eyes in the normal group did not suffer from myopia,and there was significant difference in the three groups (P ＜0.05).The average axial length of right eyes in the P6 group [(2.49 ± 0.08) mm] and the P10 group [(2.51 ±0.03)mm] was shorter than that in the normal group [(2.58 ± 0.04) mm],with significant difference (P ＜ 0.05).Immunohistochemistry showed that the expression of caspase-3 had a dramatically increase in ganglion cell layer and inner nuclear layer of retina of mice in P6 group and P10 group.TUNEL results showed that brown-stained positive apoptotic cells appeared in ganglion cell layer in the P6 and P10 group,while Western blot showed that the expression of caspase-3 protein in mouse retina in P6 group (gray value 52.70％) and P10 group (gray value 35.76％) was upregulated.Conclusion Early lighting exposure can induce premature myopia of mice,and the earlier the mice receive light,the higher the relative degree of myopia is;meanwhile during the process of premature myopia,ganglion cells and nuclear layer cells suffer apoptosis,as well as caspase-3 protein involves in the occurrence of apoptosis.
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Objective To asses stereoacuity and the factors that influence stereopsis in children after unilateral cataract extraction.Methods Sixty-two children who were diagnosed as unilateral cataract and underwent cataract extraction with intraocular lens implantation were included in this study.Data are recorded on age at presentation and the surgery,the presence of strabismus,the refractive error,and the best corrected distant visual acuity (BCDVA) of both eyes and stereoacuity.Sixty-two patients were followed up for 14 ～ 60 months.Results Sixty-two patients were divided into two groups according to stereoacuity.Thirty-one patients in group A achieved stereopsis better than 400 s of arc.Group B had 31 patients whose stereoacuity was poorer than 400 s of arc.The mean age at presentation and surgery were 4.6 ± 3.4 and 6.3 ±4.5 years in group A and 2.1 ±2.1 and 2.4 ±2.2 years in group B.51.6％ of patients in group A achieved a BCDVA of 20/40 or better,but in group B,only 6.5％ of patients achieved a BCDVA of 20/40.Those who had strabismus after cataract surgery were 6.5％ in group A and 35.5％ in group B.There was statistically significant difference in age at presentation and the surgery (t =4.03,4.53,P ＜0.01),good BCDVA(x2 =15.34,P ＜ 0.01) and absence of strabismus (x2 =7.88,P ＜ 0.01) between two groups.Conclusions Stereopsis can develop in children after pediatric unilateral cataract extraction and intraocular lens implantation.Good stereoacuity is correlated with later manifesting cataracts,absence of strabismus and good BCDVA.
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This study was aimed to develop non-toxic, high transfection efficiency polyethyleneimine(PEI) cationic nanoparticles. The exosyndrome of PEI cationic nanoparticles was measured by zeta sizer, ultraviolet and visible spectroscopy. The condensation ability and the resistance to DNaseI of pEGFP-N1/PEI and pEGFP-N1/PEI modified polyethylene glycol(PEG) were evaluated by agarose gel electrophoresis. The cell toxicity of polyethyleneimine cationic nanoparticles was measured by using MTT test. The orthogonal design was used to optimize the transfection efficiency with the N/P ratio, the grafting ratio and the gene dosage as the factors. The experimental results showed that pEGFP-N1/PEI nanoparticles have lower cell toxicity, better composite ability and better resistance to DNAseI. The highest transfection efficiency of PEI cationic nanoparticles was 91% by using the PEI nanoparticles with the N/P ratio 40:1 and gene dosages 6 microg/well. PEI cationic nanoparticle modified by PEG effectively transferred DNA to hepatoma carcinoma cells and it is a non-toxic, with high transfection efficiency, and a promising non-viral carrier for gene delivery. The transfection efficiency will be improved by optimizing the experiment condition.
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Humans , Carcinoma, Hepatocellular , Genetics , Pathology , Cell Line, Tumor , Gene Transfer Techniques , Liver Neoplasms , Genetics , Pathology , Nanoparticles , Chemistry , Polyethylene Glycols , Chemistry , Polyethyleneimine , Chemistry , Transfection , MethodsABSTRACT
AIM: To explored the potential role of HIF-1α in reducing the neuronal apoptosis and promoting the neuronal proliferation after stroke in rats. METHODS: The bone marrow-derived mesenchymal stem cells (BMSCs) were lentivirally transduced to express the stable form of HIF-1α. Ischemic stroke was induced by permanent middle cerebral artery occlusion (MCAO) in Sprague-Dawley rats. Neurological function was evaluated by modified neurological severity score (mNSS). Cerebral infarct volume was measured by TTC staining. Immunohistochemistry and terminal deoxynucleotidyltransferase mediated dUTP nick end labeling (TUNEL) method were performed to detect neuronal proliferation and apoptosis. RESULTS: Significant improvement of neurological deficits was found in BMSCs-mHIF-1α rats as compared to the control animals at 14th d and 28th d after MCAO (P<0.05). Significant reduction of infarct volume was observed in rats in BMSCs-mHIF-1α group at 3rd day after MCAO (P<0.05). Histological evaluation showed that BMSCs-mHIF-1α treatment significantly promoted neuronal survival and proliferation in the ischemic boundary area. CONCLUSION: Constitutive expression of HIF-1α in BMSCs reduces the neuronal apoptosis and promotes the neuronal proliferation after stroke in rats.
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[Objective] To investigate the effects of preconditioning with low concentration of hydrogen peroxide (H2O2) on oxidative stress-induced BMSC apoptosis.[Methods] In vitro separation,purification,culture,and amplification of bone marrow mesenchymal stem cells were performed.BMSC were insulted with 0,50,100,200,300,400,500 μmol/L H2O2 and the effect of different consentration of H2O2 on BMSC was detected by Flow cytometry (FCM).And then cells were preconditioned with different consentraion of H2O2.(FCM) was used to determine the protective role of H2O2 preconditioning on BMSC apoptosis,BMSC chromatin distribution changes were observed by Hoechst33324;BMSC Caspase-3 and Bcl-2 gene changes were detected by RT-PCR.[Results] Analysis of BMSC apoptosis by flow cytometry showed that H2O2 induced BMSC apoptosis in a dose-dependent manner,and pretreatment of the cells with low concentration of H2O2 prevented subsequent stimulation with high H2O2.RT-PCR results showed that preconditioning with low concentration of H2O2 reduced the BMSC Caspase-3 gene expression but increased Bcl-2 gene expression.[Conclusion] Preconditioning with low concentration of H2O2 has an adaptive role in BMSC,and its mechanism may be related to inhibit abnormal gene expression of Caspase-3 and increase the gene expression of Bcl-2.
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AIM To study the adaptive cytoprotection of H 2O 2 preconditioning against oxidative stress damage in PC12 cell and the relationships between brain derived neurotrophic factor (BDNF) and the adaptive cytoprotection of H 2O 2 preconditioning. METHODS The viability of PC12 cells was evaluated by MTT assay. Apoptosis was detected by PI stain flow cytometer. Expression of BDNF was analyzed by immuo flow cytometer. RESULTS After H 2O 2 preconditioning, the survival of PC12 cells exposure to H 2O 2 (20～60 ?mol?L -1 ) was increased and the apoptosis of PC12 cells induced by H 2O 2 (20 or 30 ?mol?L -1 ) was inhibited and the expression of BDNF in PC12 cells was enhanced. CONCLUSION H 2O 2 preconditioning is protective against the damage of PC12 cells induced by H 2O 2 and its mechanisms may involve up modulation of the expression of BDNF.
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Objective This paper was to observe the immediate protective effects of Radix Codonopsis,Radix Rheum,Radix Salvia and Shu Gan Tiao Wei Tang on gastric mucosas and to discuss the universality of the immediate protection of Traditional Chinese Medicine.Method The drugs were given oral to rats,then the absolute ethanol 2ml were administered immediately to observe the effects of decoction of Radix Codonopsis,Radix Rheum,Radix Salvia and Shu Gan Tiao Wei Tang on gastric mucosal damage;respectively.Results ⑴The 10g/kg of Radix Codonopsis decoction pre-injected oral inhibited markedly gastric mucosal damage with 79 9% of the inhibitory rate.⑵The 12g/kg of Radix Rheum decoction was pre-injected oral,the inhibitory rate was 48 1%.⑶When the 12g/kg of Radix Salvia decoction was pre-injected oral,the inhibitory rate was 61 53%.⑷When the 6g/kg of Shu Gan Tiao Wei Tang pre-injected was oral,the inhibitory rate was 56 84%;When the 12g/kg of Shu Gan Tiao Wei Tang was pre-injected oral,the inhibitory rate was 79 79%.Conclusions All of the Radix Codonopsis,Radix Rheum,Radix Salvia and Shu Gan Tiao Wei Tang possessed the immediate protective effect on gastric mucosas.Among the onefold drugs,the effect of Radix Codonopsis is the strongest,the Radix Salvia is secondly and the Radix Rheum is last.
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AIM:To explore the survivorship and the mechanism of the intravenous administration of bone marrow stromal stem cells(BMSCs) for treating permanent focal cerebral ischemia in rats.METHODS:After purified,proliferated,and marked with BrdU,the BMSCs were injected intravenously into rats 1 d after focal cerebral ischemia.Modified neurological severity score(mNSS) was evaluated before and following 1,7,14 and 28 d after middle cerebral artery occlusion(MCAO).Rats were executed at 1,7,14 and 28 d after MCAO.Brain sections were stained with hematoxylin and eosin(HE) for determining the infarct volume.Slides were stained by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling(TUNEL) and immunostaining for cleaved caspase-3 method for apoptosis detection and mechanism exploration in situ.RESULTS:mNSS in BMSCs-transplanted group at 14th day and 28th day of MCAO was significantly lower than that in control group(P
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AIM:To explored the potential role of HIF-1? in reducing the neuronal apoptosis and promoting the neuronal proliferation after stroke in rats. METHODS:The bone marrow-derived mesenchymal stem cells (BMSCs) were lentivirally transduced to express the stable form of HIF-1?. Ischemic stroke was induced by permanent middle cerebral artery occlusion (MCAO) in Sprague-Dawley rats. Neurological function was evaluated by modified neurological severity score (mNSS). Cerebral infarct volume was measured by TTC staining. Immunohistochemistry and terminal deoxynucleotidyltransferase mediated dUTP nick end labeling (TUNEL) method were performed to detect neuronal proliferation and apoptosis. RESULTS:Significant improvement of neurological deficits was found in BMSCs-mHIF-1? rats as compared to the control animals at 14th d and 28th d after MCAO (P
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AIM:To investigate the role of SDF-1? in migrating of bone marrow stromal cells to the injured areas. METHODS:Ischemic brain lesion model was created in rats by permanent middle cerebral artery occlusion (MCAO). 48 SD rats were divided randomly into 2 groups. Group 1:phosphate buffered saline (PBS 1 mL) for control (n=25); Group 2:BMSCs (2?106) were injected intravenously at 24 h after MCAO (n=24). After propagated in BMSCs,Ad5/F35 GFP (green fluorescent protein) was infected to BMSCs. The expression of SDF-1? (stromal cell-derived factor-1?) mRNA in the penrumbral tissue was assayed by real-time quantitative PCR. The expression of CXCR4 on MSCs was detected by flow cytometry. Confocal microscopy was used to detect the GFP-labeled MSCs migration. RESULTS:Ad5/F35 GFP signals was observed in almost infected BMSCs. The expressions of SDF-1? mRNA in the thalamus and hippocampus of the ischemic brains were peaked at 3rd day after stroke,followed by a decrease at 14th day post-ischemia. The expression of SDF-1? mRNA in the cortex of the ischemic brains was peaked at 7th day post-ischemia,still at high level at 14th day post-ischemia. The median percentage of surface CXCR4 expression in BMSCs was 14%. GFP labeled BMSCs were detected in the origination of the middle cerebral artery (olfactory area) at 6 h,after 3 days in the prenumbra tissue such as thalamus,and in the cortex more labeled cells were found after 14 d post-ischemia.CONCLUSION:BMSCs can pass through the blood brain barrier of ischemic rats. Its mechanism might be associated with the expression of SDF-1? in the ischemic brain.