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@#AIM: To explore the differentiation of bone marrow-derived mesenchymal stem cells from peripheral blood to the retina and the expression of ciliary neurotrophic factor(CNTF). We also investigate the mechanism by which Bu Shen Yi Jing Fang could treat dry age-related macular degeneration(ARMD). <p>METHODS:C57BL/6 mice were administered with sodium iodate(NaIO3)by tail intravenous injection. One day after modeling, 3×106 green fluorescent protein labeled bone marrow-derived mesenchymal stem cells(GFP-BMSCs)were injected into the tail vein. The injected mice were randomly divided into distilled water group and Bu Shen Yi Jing Fang group according to random number table, and 12 mice in each group. The mice were intragastrical administrated with either Bu Shen Yi Jing Fang solution or distilled water every day. Twelve healthy C57BL/6 mice were fed regularly as the normal group. At 14d after the treatment, the differentiation of GFP-BMSCs in retina was determined by immunofluorescence, and the expression of CNTF in the retina was detected by immunofluorescence and quantitative real-time PCR.<p>RESULTS: Immunofluorescence staining showed that there were more glial fibrillary acidic protein(GFAP)and GFP double-stained positive cells in the Bu Shen Yi Jing Fang group than in the distilled water group(<i>P</i><0.01), and the positive rate of retinal pigment epithelium 65(RPE65)was not significantly different between two groups(<i>P</i>>0.05). There were no Rhodopsin and GFP double-stained positive cells in the two groups. Immunofluorescence and quantitative real-time PCR showed that the expression of CNTF in the Bu Shen Yi Jing Fang group was higher than which in the distilled water group(<i>P</i><0.05). <p>CONCLUSION: Bu Shen Yi Jing Fang facilitated the differentiation of peripheral blood stem cells into glial cells in the retina and the expression of CNTF, which might be one of the mechanisms of Bu Shen Yi Jing Fang in the treatment of dry ARMD.
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In the central nervous system (CNS), three types of myelin-associated inhibitors (MAIs) have major inhibitory effects on nerve regeneration. They include Nogo-A, myelin-associated glycoprotein, and oligodendrocyte-myelin glycoprotein. MAIs possess two co-receptors, Nogo receptor (NgR) and paired immunoglobulin-like receptor B (PirB). Previous studies have confirmed that the inhibition of NgR only results in a modest increase in regeneration in the CNS; however, the inhibitory effects of PirB with regard to nerve regeneration after binding to MAIs remain controversial. In this study, we demonstrated that PirB is expressed in primary cultures of retinal ganglion cells (RGCs), and the inhibitory effects of the three MAIs on the growth of RGC neurites are not significantly decreased after direct PirB knockdown using adenovirus PirB shRNA. Interestingly, we found that retinal Müller cells expressed PirB and that its knockdown enhanced the regeneration of co-cultured RGC neurites. PirB knockdown also activated the JAK/Stat3 signaling pathway in Müller cells and upregulated ciliary neurotrophic factor levels. These findings indicate that PirB plays a novel role in retinal Müller cells and that its action in these cells may indirectly affect the growth of RGC neurites. The results also reveal that PirB in Müller cells affects RGC neurite regeneration. Our findings provide a novel basis for the use of PirB as a target molecule to promote nerve regeneration.
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In the central nervous system (CNS), three types of myelin-associated inhibitors (MAIs) have major inhibitory effects on nerve regeneration. They include Nogo-A, myelin-associated glycoprotein, and oligodendrocyte-myelin glycoprotein. MAIs possess two co-receptors, Nogo receptor (NgR) and paired immunoglobulin-like receptor B (PirB). Previous studies have confirmed that the inhibition of NgR only results in a modest increase in regeneration in the CNS; however, the inhibitory effects of PirB with regard to nerve regeneration after binding to MAIs remain controversial. In this study, we demonstrated that PirB is expressed in primary cultures of retinal ganglion cells (RGCs), and the inhibitory effects of the three MAIs on the growth of RGC neurites are not significantly decreased after direct PirB knockdown using adenovirus PirB shRNA. Interestingly, we found that retinal Müller cells expressed PirB and that its knockdown enhanced the regeneration of co-cultured RGC neurites. PirB knockdown also activated the JAK/Stat3 signaling pathway in Müller cells and upregulated ciliary neurotrophic factor levels. These findings indicate that PirB plays a novel role in retinal Müller cells and that its action in these cells may indirectly affect the growth of RGC neurites. The results also reveal that PirB in Müller cells affects RGC neurite regeneration. Our findings provide a novel basis for the use of PirB as a target molecule to promote nerve regeneration.
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@#AIM: To establish the model of bone marrow mesenchymal stem cells(BMSCs)over-expressing ciliary neurotrophic factor(CNTF)by applying adenovirus. It may provide novel strategies for <i>in-vitro </i>investigations of retinopathies.<p>METHODS: GFP-adenovirus and CNTF-adenovirus were synthesized then used to transfect BMSCs of passage 3. Blank control group(without adenovirus transfected group), negative control group(GFP-adenovirus transfected group), and experimental group(CNTF-adenovirus transfected group)were included in this study. On the 1, 2, 3d post-transfected, ELISA assay was applied to examine CNTF protein-secretion in the supernate.<p>RESULTS: GFP-adenovirus and CNTF-adenovirus models were successfully established. The CNTF protein levels in the supernate were higher in experimental group than those in the blank control group and negative control group(<i>P</i><0.05). <p>CONCLUSION: CNTF-modified BMSCs by adenovirus could efficiently secrete CNTF protein <i>in-vitro</i>.
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Transient receptor potential vanilloid subtype 1 (TRPV1) on astrocytes prevents ongoing degeneration of nigrostriatal dopamine (DA) neurons in MPP⁺-lesioned rats via ciliary neurotrophic factor (CNTF). The present study determined whether such a beneficial effect of astrocytic TRPV1 could be achieved after completion of injury of DA neurons, rather than ongoing injury, which seems more relevant to therapeutics. To test this, the MPP⁺-lesioned rat model utilized here exhibited approximately 70~80% degeneration of nigrostriatal DA neurons that was completed at 2 weeks post medial forebrain bundle injection of MPP⁺. TRPV1 agonist, capsaicin (CAP), was intraperitoneally administered. CNTF receptor alpha neutralizing antibody (CNTFRαNAb) was nigral injected to evaluate the role of CNTF endogenously produced by astrocyte through TRPV1 activation on DA neurons. Delayed treatment of CAP produced a significant reduction in amphetamine-induced rotational asymmetry. Accompanying this behavioral recovery, CAP treatment increased CNTF levels and tyrosine hydroxylase (TH) activity in the substantia nigra pars compacta (SNpc), and levels of DA and its metabolites in the striatum compared to controls. Interestingly, behavioral recovery and increases in biochemical indices were not reflected in trophic changes of the DA system. Instead, behavioral recovery was temporal and dependent on the continuous presence of CAP treatment. The results suggest that delayed treatment of CAP increases nigral TH enzyme activity and striatal levels of DA and its metabolites by CNTF endogenously derived from CAP-activated astrocytes through TRPV1, leading to functional recovery. Consequently, these findings may be useful in the treatment of DA imbalances associated with Parkinson's disease.
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Animals , Rats , Antibodies, Neutralizing , Astrocytes , Capsaicin , Ciliary Neurotrophic Factor , Dopamine , Dopaminergic Neurons , Medial Forebrain Bundle , Models, Animal , Neurons , Parkinson Disease , Pars Compacta , Receptor, Ciliary Neurotrophic Factor , Tyrosine 3-MonooxygenaseABSTRACT
Age-related macular degeneration(AMD) is the major cause of irreversible damage to vision in over 60 years old.Its incidence is increasing with age.Dry AMD is one of the most common types,causing the loss of vision is very slow.At the late stage,the geographic atrophy will be found,in which central vision is severely lost.It is widely believed that chronic inflammantion injury,oxidative stress injury,lipofuscin,drusen and choroid ischemia are the major pathogenesis.Now,the treatment is major aiming to the pathogenesis.But some new therapies such as crocetin,curcumin,ciliary neurotrophic factor,nanoceria,human monoclonal antibody,photoreceptor and stem cell transplantation have some efficiency to dry AMD.This article reviews the research advances in therapy of dry AMD.
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@#Ciliary neurotrophic factor(CNTF)is one of the most studied neurotrophic factors. Encapsulated cell technology provides a safe and effective route for the clinical application of CNTF. A large body of evidence shows that CNTF has a neuroprotective effect on glaucoma, age-related macular degeneration, optic nerve injury and other eye diseases. This review focuses on the potential clinical application of CNTF in eye diseases.
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Objective To observe the effects of Jiaji electroacupuncture and neurodynamic mobilization technique on axon regeneration and content of brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) in rabbits with sciatic nerve injury. Meth-ods 30 male adult rabbits were assigned to sham (A, n=6), model (B, n=6), neurodynamic mobilization (C, n=6), Jiaji electroacupuncture (D, n=6), and Jiaji electroacupuncture combine with neurodynamic mobilization (E, n=6) groups. The group C was treated with neurodynam-ic mobilization, the group D with Jiaji electroacupuncture, and the group D with both 3 days after modeling of clamping at sciatic nerve, while the groups A and B with no treatment. The axon regeneration was observed with HE staining, and the content of BDNF and CNTF in serum was measured with ELISA 4 weeks after treatment. Results The axons regeneration was observed better in the groups C, D and E than in the group B. The content of BDNF and CNTF was more in the groups C, D and E than in the group B (P<0.05). Conclusion Both Ji-aji electroacupuncture and neurodynamic mobilization can improve axon regeneration with synergistic action, which may associate with the increase of BDNF and CNTF in serum.
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Objective To investigate the effect of Trillium tschonoskii Maxim extract on the expression of ciliary neurotrophic factor (CNTF) and its receptor (CNTFRα) after spinal cord injury in rats. Methods Forty-five rats were equally and randomly divided into control group (group A), model group (group B) and Trillium tschonoskii Maxim treated group (group C). Allen's weight drop method was used to reproduce acute spinal cord injury (SCI) model in rats of the group B and C. In group C, the rats were gavaged with Trillium tschonoskii Maxim extract 2 weeks before the injury, while rats in group A and B were fed a same quantity of distilled water.1, 7 or 14 days after injury, the rats were sacrificed to observe the structure of nerve cells after HE and Nissl staining, and the expression of CNTF and CNTFRα with immunohistochemical method, RT-PCR and Western blotting. Results HE staining showed that the structure of spinal cord in the the rats group C was more discernible, with milder edema and necrosis of nerve cells, as compared with that of group B. Nissl staining showed that Nissl bodies were decreased or disappeared in anterior horn motor neurons in both group B and C, but it was significantly less marked in group C than that in group B. Immunohistochemical staining, Western blotting and RT-PCR revealed that the protein and mRNA of CNTF and CNTFRα were positively expressed in rats of every group. The mRNA levels of CNTF and CNTFRα in group C were higher than those in group B. Conclusions Extract of Trillium tschonoskii Maxim can up-regulate the expression of CNTF and CNTFRα, and plays a protective role against injury to spinal cord.
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@#Objective To observe the effects of Jiaji electroacupuncture and neurodynamic mobilization technique on axon regeneration and content of brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) in rabbits with sciatic nerve injury. Methods 30 male adult rabbits were assigned to sham (A, n=6), model (B, n=6), neurodynamic mobilization (C, n=6), Jiaji electroacupuncture (D, n=6), and Jiaji electroacupuncture combine with neurodynamic mobilization (E, n=6) groups. The group C was treated with neurodynamic mobilization, the group D with Jiaji electroacupuncture, and the group D with both 3 days after modeling of clamping at sciatic nerve, while the groups A and B with no treatment. The axon regeneration was observed with HE staining, and the content of BDNF and CNTF in serum was measured with ELISA 4 weeks after treatment. Results The axons regeneration was observed better in the groups C, D and E than in the group B. The content of BDNF and CNTF was more in the groups C, D and E than in the group B (P<0.05). Conclusion Both Jiaji electroacupuncture and neurodynamic mobilization can improve axon regeneration with synergistic action, which may associate with the increase of BDNF and CNTF in serum.
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Objective To study whether bellidifolin has rehabilitation effect on injured sciatic nerve rats,and whether ciliary neurotrophic factor (CNTF) is involved in this mechanism.Methods 225 male wistar rats were made to be sciatic nerve injured models,with right sciatic nerve being cut and sewed under microscopy,left sciatic being sham.Rats were randomly divided into control group,bellidifolin 25 mg,50 mg,75 mg groups and mecobalamin group,with 45 rats in each group.Rats in control group were just injected sodium chloride,others were intra-peritonealiy injected different doses of bellidifolin and mecobalamin after operation.Results Three weeks after operation,SFl of control group,mecobalaming group,bellidifolin 25 mg,50 mg and 75 mg group were-84.35± 4.87,-45.20±2.30,-70.42±4.21,-57.73±3.46 and-64.38±4.38 respectively.Compared with control group,others showed significant differences (P＜0.05).There were statistically differences between bellidifolin groups and mecobalaming group(P＜0.05).Within bellidifolin groups,50 mg group showed difference compared with 25 mg and 75 rg groups(P=0.031).TSW results also showed differences among bellidifolin groups,control group and mecobalaming group.There were statistical differences among bellidifolin groups(P＜0.05).Each groups with immunohistochemistry analysis,CNTF expression showed statistically differences among bellidifolin 50 mg group and 25 mg,70 rg groups,Bellidifolin 50 mg group was higher than others(P＜0.05).Conclusion Bellidifolin can promote the recovery of injured sciatic nerve,especially the concentration of 50 mg bellidifolin,and CNTF is involved in the rehablitation process.
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Objective To study the rehabilitation effect of Bellidifolin for injuried sciatic nerve,and to explore whether ciliary neurotro-phic factor ( CNTF) is involved in this mechanism. Methods The right sciatic nerver of 225 male wistar rats was cut and sewed under mi-croscopy. Rats were devided into 5 groups,as control group,Bellidifolin 25 mg group,50 mg group、75 mg group and Mecobalamin group. The control group were injected sodium chloride,other groups were injected different dose of Bellidifolin and Mecobalamin. 1,3 and 5 weeks later, the motor nerve conduction velocity( MNVC) and gastrocnemius muscle cross-sectional area were detected,CNTF positive area were analysed by immunohistochemical method. Results There were differences among bellidifolin groups,control group and mecobalamin group in Nerve conduction velocity. Within Bellidifolin groups,50 mg group compared with 25 mg and 75 mg groups,there were statistically differences( P=0. 025). Three weeks after operation,gastrocnemius muscle cross-sectional area of control group,mecobalaming grop and Bellidifolin 25 mg group,50 mg group,and 75 mg group were(455. 06 ± 29. 38),(679. 03 ± 81. 48),(465. 31 ± 71. 55),(670. 24 ± 91. 26) and (669. 28 ± 78. 54) respectively,compared with control group and Bellidifolin 25 mg group,others had a significant difference(P<0. 05). CNTF expres-sion showed billidifolin 50 mg group are higher than others(P<0. 05). Conclusion Bellidifolin can improve the rehabilitation of injured sciatic nerve. CNTF is involved in this mechnism.
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BACKGROUND:Conventional ophthalmic delivery of ciliary neurotrophic factor (CNTF) is extremely difficult to pass the blood-retinal barrier, resulting in a very low bioavailability and the need of long-term drug delivery. To solve the problem, the CNTF can be encapsulated in a semi-permeable membrane to form a microcapsule, which may then achieve the release of bioactive substances encapsulated, or bioactive molecules secreted by living cels and smal molecular metabolites through semi-permeability of the special membrane. OBJECTIVE:To prepare a special structural CNTF sustained-release microcapsule. METHODS: A selected poly(ether sulfone) holow fiber was cut into 1 cm long with its two ends sealed by 1181-M medical adhesive using UV curing. To prepare CNTF encapsulated microcapsule, one end was first sealed, and then the CNTF was loaded to poly(ether sulfone) microcapsule from the other end which then was sealed. The leaching liquor of sustained-release microcapsule was co-cultured with mouse fibroblast L929, to observe the cytotoxicity of the microcapsule. The sustained-release microcapsule was co-cultured with mouse retinal pigment epithelial cels, to observe the celladhesion ability of the microcapsule. The CNTF sustained-release microcapsule was immersed in physiological saline, to observe the degradability. Moreover,in vitro release behavior of immunoglobulin and CNTF were evaluated. RESULTS AND CONCLSION:The CNTF sustained-release microcapsule had an inner diameter of about 398 μm and a membrane thickness of about 145 μm. The microcapsule presents a lot of macropores in the outer wal and many 10 nanometers micropores in the inner wal. The sustained-release microcapsule was not degraded in saline within 4 months, indicating good cellcompatibility. The microcapsule can selectively release CNTF while protecting against invading of antibodies (IgG), showing its good selective permeability. Meanwhile, the sustained-release microcapsule improved the initial burst release of traditional drug delivery vesicles. The microcapsule presents a mild sudden release in the middle stage, and then a sustained release.
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Background The application of mesenchymal stem cells to transfer specific genes is under investigation in various diseases.Using this strategy may provide a more effective method to supply exogenous neurotrophic factors to the cental nervous system,including retina.Objective This study was to construct ciliary neurotrophic factor(CNTF)-overexpressing bone marrow mesenchymal stem cells (BMSCs) using lentiviral vectors.Methods Rat secreted-CNTF gene cDNA was synthesized and subcloned into a lentiviral vector plasmid pHⅣ-dTomato to construct recombinant vector CNTF-dTomato.CNTF-dTomato/pH Ⅳ-dTomato plasmid were co-transfected into 293T packaging cell line with packaging plasmid psPAX2 and enveloped plasmid pMD.2G to produce recombinant lentivirus CNTF-lenti and control-lenti.Rat BMSCs were infected with CNTF-lenti/control-lenti to produce CNTF-BMSCs and control-BMSCs.Expression of dTomato and efficiency of infection was evaluated under the fluorescence microscope.Uninfected BMSCs(pure BMSCs) served as the blank control.CNTF protein level in the supernate was detected by enzyme-linked immunosorbent assay (ELISA) and compared among the blank-BMSCs group,control-BMSCs group and CNTF-BMSCs group.Adipogenic and osteogenic differentiation of CNTF-BMSCs were induced using adipogenic-inducible medium and osteogenic-inducible medium and identified using oil-red O staining and alizarin red S (ARS) staining.Results After CNTF-dTomato plasmid was transfected into Stbl3 competent cells,the colony PCR product was 1 033 bp.The inserted sequence in the pHⅣ-dTomato plasmid was coincident with the expected one.The results of DNA sequencing showed that CNTF-dTomato plasmid was successfully constructed.The infection rate of CNTF-lenti was approximately 95％.ELISA showed that on the post-infected day 2,3,7,the CNTF protein levels in the supernate were significantly higher in the CNTF-BMSCs group than those in the blankBMSCs group and control-BMSCs group (all at P=0.000).In the CNTF-BMSCs group,the CNTF protein levels in the supernate were significantly increased on post-infected day 2,3,7 compared with day 1 (P =0.013,0.004,0.042).The adipogenic-induced cells showed the red staining to oil red O,and osteogenic-induced cells presented the orange staining to ARS.Conclusions BMSC line with stable expression of CNTF is successfully established by lentiviral vectors.CNTF-BMSCs have the potential to differentiate towards adipocytes and osteoblasts.
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PURPOSE: To investigate neurotrophins expression and neurotrophic effect change in mesenchymal stem cells (MSCs) under different types of stimulation. METHODS: Rats were exposed in 10,000 lux white light to develop light-induced retinal injury. Supernatants of homogenized retina (SHR), either from normal or light-injured retina, were used to stimulate MSCs. Quantitative real time for polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were conducted for analysis the expression change in basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) in MSCs after stimulation. Conditioned medium from SHR-stimulated MSCs and control MSCs were collected for evaluation their effect on retinal explants. RESULTS: Supernatants of homogenized retina from light-injured rats significantly promoted neurotrophins secretion from MSCs (p<0.01). Conditioned medium from mesenchymal stem cells stimulated by light-injured SHR significantly reduced DNA fragmentation (p<0.01), up-regulated bcl-2 (p<0.01) and down-regulated bax (p<0.01) in retinal explants, displaying enhanced protective effect. CONCLUSIONS: Light-induced retinal injury is able to enhance neurotrophins secretion from mesenchymal stem cells and promote the neurotrophic effect of mesenchymal stem cells.
OBJETIVO: Investigar a expressão de neurotrofinas e mudança no efeito neurotrófico de células-tronco mesenquimais (MSCs) sob diferentes tipos de estimulação. MÉTODOS: Os ratos foram expostos em 10.000 lux de luz branca para desenvolver a lesão da retina induzida por luz. Os sobrenadantes de homogeneizado de retina (SHR) quer a partir de retina normal ou da lesada por luz, foram usados para estimular as células-tronco mesenquimais. O RT-PCR quantitativa e ELISA foram realizados para análise da alteração de expressão do fator básico de crescimento de fibroblastos (bFGF), do fator neurotrófico derivado do cérebro (BDNF) e do fator neurotrófico ciliar (CNTF) em MSCs após a estimulação. O meio condicionado de células-tronco mesenquimais estimuladas por SHR e controles foram coletadas para avaliação de seu efeito sobre os explantes de retina. RESULTADOS: SHR de retinas de rato lesadas por luz promoveram aumento significativo de secreção de neurotrofinas em MSCs (p<0,01). O meio condicionado de SHR lesado por luz reduziu significativamente a fragmentação do DNA de MSCs (p<0,01), elevação de Bcl-2 (p<0,01) e redução de bax (p<0,01) em explantes de retina, mostrando um aumento do efeito protetor. CONCLUSÕES: A lesão da retina induzidos pela luz é capaz de aumentar a secreção de neurotrofinas e promover o efeito neurotrófico de células-tronco mesenquimais.
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Animals , Rats , Light , Mesenchymal Stem Cells , Nerve Growth Factors , Retina/radiation effects , Brain-Derived Neurotrophic Factor , Cells, Cultured , Ciliary Neurotrophic Factor , Disease Models, Animal , Fibroblast Growth Factors/metabolism , Mesenchymal Stem Cells/cytology , Random Allocation , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Retina/cytology , Retina/injuriesABSTRACT
Background Researches demonstrated that ciliary neurotrophic factor (CNTF) can enhance survival and promote differentiation of neutron.Meanwhile,CNTF also is thought to play an important role in the development of visual pathway.But,less studies are reported in the relationship of CNTF and form deprivation amblyopia.Objective This study was to investigate the expressions of CNTF in visual cortex area 17 in form deprivation amblyopia model.Methods Twelve 4-week-old cats were randomized into normal group and form deprivation amblyopia group.Monocular form deprivation amblyopic models were established in 6 cats by eyelids suture method.Pattern visual evoked potential(P-VEP) was recorded to evaluate the amblyopic models 16 weeks later following the eyelids suturing.Then,bilateral visual cortex tissue was incised at a vertical in sagittal axis fashion to prepare the section.Nissl staining was used to detect the morphologies of neurons.Expression of CNTF in Ⅰ-Ⅵ layers of visual cortex area 17 was located and quantified by immunochemistry.The positive cell number and gray scale for CNTF were calculated and compared between two groups.The use of the animals complied with Regulations for the Administration of Affairs Coucerning Experimental Animals by State Science and Technology Commission.Results Compared with the normal group,P-VEP amplitude was significantly reduced (6.11 ±1.56 μV vs.11.42±t.92 μV) and latency was significantly prolonged(75.77±9.83 ms vs.58.56±7.17 ms) in the form deprivation amblyopia group (t=5.272,3.464,P＜0.05).Nissl staining showed that the number of neurons in the form deprivation amblyopia group was less than that in the normal group.In the form deprivation amblyopia group,neurons became shrinkage and turned round,cytoplasmic processes get shortened,and the nucleus became small.The number of Nissl bodies was decreased.lmmunochemistry showed the positive neutrons for CNTF in Ⅰ-Ⅵ layers of visual cortex area 17 in hoth normal cats and model cats with the more positive cells in Ⅱ-Ⅳ layers.Compared with the normal group,the positive cell number for CNTF was significantly reduced in Ⅱ-Ⅳ layers of visual cortex area 17 in the form deprivation amblyopia group (Ⅱ layer:95.93±8.24 vs.116.25±6.52;I layer:102.65±7.45 vs.125.23±8.21;Ⅳ layer:l10.65±6.85 vs.139.54±4.26) (t=4.737,4.989,8.773,P＜0.05).In addition,the gray scale of CNTF positive cells was significantly lower in Ⅱ-Ⅳ layers of visual cortex area 17 in the form deprivation amblyopia group than that the normal group (Ⅱ layer:106.98 ± 8.86 vs.138.65 ± 6.38 ; Ⅲ layer:109.56 ± 8.69 vs.149.59 ±8.55;Ⅳ layer:l16.65 ±9.52 vs.155.76±9.87) (t=7.105,8.043,6.986,P＜0.05).Both CNTF positive cell number and gray scale in Ⅰ,Ⅴ,Ⅵ layers of visual cortex area 17 had no significant differences between two groups (P＞0.05).Conclusions Form deprivation in critical period of a new born animal may lead to distributing abnormality of CNTF in visual cortex,which maybe play a role in the development of form deprived amblyopia.
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Background The key premise of genetic research and treatment is to select the desired gene vector,ultrasound microbubbles as a new type of gene vector can safe,fast and effectively enhance the gene transfection and expression by reversibility increasing the permeability of cells. Objective This study was to observe the effects of ciliary neurotrophic factor(CNTF)gene mediated by ultrasound microbubbles intraocular transfer on visual function and retinal ganglion cell(BGCs)after optic nerve injury. Methods Fifty-five adult Sprague-Dawley(SD)rats were divided into 6 groups randomly,including normal control group(n=5),sham injury group(n=10),simple injury group(n=10),naked plasmid group(n=10),plasmid+ultrasonic irradiation group(n=10)and ultrasound microbubbles group(n=10).The model of optic nerve injury Was made by forceps clip on the fight optic nerve.and the corresponding intervene was performed in different groups.Flash visual evoked potential(F
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Ciliary neurotrophic factor (CNTF) is well known as a growth/survival factor of neuronal tissue. We investigated the expression of CNTF and its specific receptor alpha (CNTFRalpha) in a unilateral ureteral obstruction (UUO) model. Complete UUO was produced by left ureteral ligation in Sprague-Dawley rats. The animals were sacrificed on days 1, 3, 5, 7, 14, 21, and 28 after UUO. The kidneys were fixed, and processed for both immunohistochemistry and in situ hybridization. CNTF immunoreactivity in sham-operated kidneys was observed only in the descending thin limb (DTL) of the loop of Henle. In UUO kidneys, CNTF expression was induced in the S3 segment (S3s) of the proximal tubule from day 1, and progressively expanded into the entire S3s and a part of the convoluted proximal tubules, distal tubules (DT), and glomerular parietal epithelium up to day 7. Upregulated CNTF expression was maintained to day 28. From day 14, the inner medullary collecting duct showed weak CNTF immunoreactivity. The CNTFRalpha mRNA hybridization signal in sham-operated kidneys was weakly detected in the DTL, DT, medullary thick ascending limb, and in a few S3s cells. After UUO, CNTFRalpha mRNA expression increased progressively in both the renal cortex and the medulla up to day 7 and increased expression was maintained until day 28. The results suggest that the S3s may be the principal induction site for CNTF in response to renal injury, and that CNTF may play a role in chronic renal injury.
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Animals , Chimera , Ciliary Neurotrophic Factor , Ciliary Neurotrophic Factor Receptor alpha Subunit , Epithelium , Extremities , Immunohistochemistry , In Situ Hybridization , Kidney , Ligation , Loop of Henle , Neurons , Rats, Sprague-Dawley , RNA, Messenger , Ureter , Ureteral ObstructionABSTRACT
@#Objective To explore oropharyngeal swallowing disorders with videofluoroscopic swallowing study (VFSS). Methods 16 patients with dysphagia accepted VFSS with 10 ml of thin barium meal (50% w/v), thick barium meal (270% w/v), biscuit coated with thick barium meal in single swallow. Their swallowing function was observed on the lateral and anterior/posterior planes, including: symmetry of pyriform sinuses, oral transit time, presence of pharyngeal delay, pharyngeal transit time, oral and pharyngeal residue, and presence of aspiration.Results 5 patients demonstrated oral swallowing disorder. 3 patients demonstrated pharyngeal swallowing disorders, that was pharyngeal delay which caused in aspiration after swallowing. 8 patients demonstrated oropharyngeal swallowing disorders, and 3 of them presented aspiration,2 patients were silent aspirators, 1 was aspiration before and 1 after swallowing. The aspiration time could not be judged from the videofluoroscopy in the other one. For 4 patients with aspiration, 3 were severe, with more than 25% of the bolus aspirated, and 1 aspirated less than 5%. Conclusion VFSS can be helpful to plan individual rehabilitation.
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Aim To determine TAT-Tcntf penetration ability and to investigate the effects of the fusion protein on SH-SY5Y cells against toxicity induced by β-amyloid peptide 25-35(Aβ_(25-35) ).Methods The conjugate(TAT-tCNTF)of TAT(47-57)of HIV-1 and the truncated human CNTF active fragment was genetic engineered and expressed in E.Coli.Immunofluorescence was used to identify cell permeation ability across membrane.MTT assay was used to measure the survival of SH-SY5Y cells injured by Aβ_(25-35).And Hoechst 33342/PI double staining was used to observe the morphology of cell apoptosis and necrosis.LDH was measured by spectrophotometric method.Results The expression vector of pBV220-TAT-tCNTF was constructed successfully.Western blot showed the recombinant fusion protein could bind specifically with CNTF antibody.The immunofluorescence assay clearly demonstrated that TAT-tCNTF did penatrate into the cells while little rhCNTF pass across the cells.Double staining and LDH release assay demonstrated that TAT-tCNTF could promote significantly the survival of the cells.Conclusion sTAT-tCNTF with high activities and effective transmembrane ability is obtained for the first time.The fusion protein protects SH-SY5Y cells from death after Aβ25-35 exposure.