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Myopia has become a major public health problem affecting the eye health of our nationals, especially adolescents, and is listed as one of the three major diseases in the world. The etiology of myopia is complex and it is currently considered to be mainly related to environmental factors and genetic factors. Dietary factors are less concerned, but it has been reported that there is a correlation between high-sugar diet and the occurrence and development of myopia. Studies have found that high-sugar diet can cause hyperinsulinemia, which mediates the increase of retinal insulin-like growth factor(IGF-1, IGF-2)expression level, and the decrease of insulin-like growth factor binding protein-3(IGFBP-3)expression level in scleral fibroblasts, thereby activating signal transduction and transcriptional activator protein 3(STAT3)signaling pathway, and then acting on scleral matrix metalloproteinases(MMP)/transforming growth factor-β(TGF-β)and other factors, resulting in a decrease in scleral extracellular matrix(ECM), eventually leading to scleral remodeling, eye axis growth and myopia. This article explores the relationship between high-sugar diet and the occurrence and development of myopia from the aspects of epidemiology and biological mechanisms by consulting relevant literature, and provides research directions for the prevention and control of myopia.
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A brand-new class of photoreceptors has been identified in the past 20a: intrinsically photosensitive retinal ganglion cells(ipRGC). With melanopsin as its photopigment, ipRGCs transmit light signals to non-imaging brain regions like the suprachiasmatic nucleus(SCN)and the olivary pretectal nucleus(OPN)to regulate circadian photoentrainment and pupillary light reflex; a small portion of the signals are projected to brain imaging regions like the dorsal lateral geniculate nucleus(dLGN)and superior colliculus(SC), to participate in imaging vision. There are six different ipRGC subtypes(M1~M6), each with its own morphological and physiological characteristics. In addition to receiving signaling inputs from the rods and cones, ipRGCs also regulate retinal signals through chemical and electrical synapses and play important roles in visual signaling and visual development. It has been discovered that ipRGCs are implicated in several systemic and ocular illnesses. Overall, various aspects of ipRGC are reviewed including the discovery, general physiological properties, signaling, and the relationship with disease in this work.
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The occurrence and development of many eye diseases are closely related to genetic and environmental factors, among which epigenetic modification is an important bridge connecting genetic and environmental factors. It can affect the levels of related genes by influencing gene transcription or translation, thereby playing a role in the pathogenesis of ocular diseases. DNA methylation is an important part of epigenetic modification which is usually regulated by three processes: de novo methylation, maintenance methylation, and demethylation, and plays an essential role in regulating gene expression. At present, researchers have conducted that DNA methylation plays an important role in repair of damage to corneal endothelium, mitochondrial dynamics regulation and diabetic retinopathy, oxidative stress response and cataracts and other eye diseases, providing new ideas in the treatment of related ocular diseases. This study presented a brief review of the role of DNA methylation in the development of related ocular diseases and provided new perspectives and directions for the screening, diagnosis, and treatment of eye diseases.
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OBJECTIVE@#To explore the underlying mechanism of inhibition by Jinkui Shenqi Pills (JKSQP) on glucocorticoid-enhanced axial length elongation in experimental lens-induced myopia (LIM) guinea pigs.@*METHODS@#Sixty 2-week old male guinea pigs were randomly divided into 4 groups with 15 guinea pigs in each group, according to the random numbers generated by SPSS software: control, LIM, saline and JKSQP groups. The control group includes animals with no treatment, while the guinea pigs in the other 3 groups received lens-induced myopization on the right eyes throughout the experiment (for 8 weeks). The saline and JKSQP groups were given daily intraperitoneal injections of 10 mg/kg hydrocortisone for 2 consecutive weeks at the same time, and then orally administered either saline or JKSQP [13.5 g/(kg•d) for 6 consecutive weeks. Body weight, anal temperature and animal appearance were observed and recorded to evaluate the GC-associated symptoms. The ocular parameters, including refraction and axial length, were measured by streak retinoscopy and A-scan ultrasonography, respectively. The levels of plasma hormones associated with the hypothalamic-pituitary-adrenal axis (HPAA), including free triiodothyronine, free thyroxine, estradiol and testosterone, were measured by radioimmunoassay, and cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate were measured by enzyme-linked immunosorbent assay. In addition, the mRNA and protein expressions of retinal amphiregulin (AREG) was measured by quantitative real-time polymerase chain reaction and Western blotting, respectively.@*RESULTS@#JKSQP effectively increased body weight and anal temperature, improved animal appearance and suppressed axial length elongation in glucocorticoid-enhanced myopic guinea pigs with normalization of 4 HPAA-associated plasma hormones (all P<0.05). The plasma level of cAMP was significantly increased, whereas the plasma level of cGMP and the mRNA and protein expressions of retinal AREG were decreased after treatment with JKSQP (all P<0.05).@*CONCLUSION@#JKSQP exhibited a significant inhibitory effect on axial length elongation with decreased expression of AREG in the retina, and normalized 4 HPAA-associated plasma hormones and the expression of cAMP and cGMP in GC-enhanced myopic guinea pigs.
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Masculino , Animales , Cobayas , Glucocorticoides , Sistema Hipotálamo-Hipofisario , Sistema Hipófiso-Suprarrenal , Miopía/metabolismo , Peso Corporal , ARN Mensajero , Modelos Animales de EnfermedadRESUMEN
Cataract is one of the most common causes of vision loss and even blindness in patients, and surgery is a proven and effective treatment option. Traditional cataract surgery for vision loss has increasingly given way to refractive cataract surgery as science and technology have progressed. There are also a variety of refractive intraocular lenses on the market place. Patients are increasingly accepting and recognizing multifocal intraocular lens(MIOL)as an alternative to traditional single focus intraocular lens(SIOL). Through classification and listing, the existing MIOL are discussed in this article, as well as the features of different types of MIOL and techniques for evaluating the clinical impacts of patients after surgery, so as to provide references for ophthalmologists.
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In recent years, fiber photometry has been widely used in the field of neuroscience as an important technique for recording the activity of neurons in the specific nuclei of freely moving animal. This review summarized the application of single-channel, multi-channel, and multi-color fiber photometry techniques in the neuroscience research of cognition, behavior, psychology and neurological diseases. In addition, it briefly introduced the applications of fiber photometry combined with functional magnetic resonance imaging technology, and fiber photometry combined with probe technology in the neuroscience research.
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Animales , Imagen por Resonancia Magnética , Neuronas , FotometríaRESUMEN
Objective:To observe the clinical efficacy of modified Yihuo Qnggan Tuiyi Tang on herpes simplex keratitis with liver meridian and wind-heat,so as to explore its mechanism of action. Method:Totally 130 patients with herpes simplex keratitis with liver meridian and wind-heat were randomly divided into control group and observation group,with 65 cases in each group. The control group was treated with conventional western medicine,while the observation group was treated with modified Yihuo Qinggan Tuiyi Tang in addition to the therapy of the control group for 4 weeks. The clinical efficacy,traditional Chinese medicine(TCM) syndrome score,symptoms and signs,adverse reactions and recurrence rate of the two groups were compared. The levels of serum interferon gamma(IFN-γ),interleukin-6(IL-6),interleukin-8(IL-8) inflammatory factors,malondialdehyde (MDA),antioxidant activity (TAC),glutathione peroxidase(GSH-PX)of oxidative stress indexes were detected by enzyme linked immunosorbent assay (ELISA). The levels of CD4+ and CD8+ were measured by flow cytometry. The recurrence rate was observed during 1-year follow-up visit. Result:The total effective rate of observation group was better than that of control group (Z=2.161,P<0.05). After treatment,the scores of eye pain,shame,photophobia,tears,vision loss,redness and total eyelid swelling were significantly lower than those of control group (P<0.05). The total effective rate of TCM syndrome in the observation group was better than that of control group (Z=2.540,P<0.05). After treatment,the scores of pain,eyelid redness,conjunctival congestion,corneal punctate pannus,corneal pannus and corneal pannus depth of symptoms and signs in observation group were better than those of control group (P<0.05). The level of serum IFN-γ in observation group was higher than that of control group,and the levels of IL-6 and IL-8 were lower than those of control group (P<0.05). Levels of MDA in the observation group was higher than that of control group (P<0.05). The levels of CD4+and CD4+/CD8+ in observation group were higher than those of control group,and the levels of CD8+ were lower than that of control group (P<0.05). The recurrence rate in observation group was significantly lower than that in control group after withdrawal for 6 months and 12 months (P<0.05). Conclusion:In addition to the therapy of routine western medicine, modified Yihuo Qinggan Tuiyi Tang has a good clinical efficacy in treating herpes simplex keratitis with liver meridian and wind-heat,and can reduce the scores of TCM syndromes,alleviate symptoms and signs,and reduce the recurrence rate. Its mechanism may be related to reducing inflammation reaction,and improving oxidative stress index and immune function.
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·Rodents have been used widely in the research of eye diseases to study visual function in animal models. Two methods of visual acuity testing in animals have been internationally recognized:the electrophysiological visual acuity test and the behavioral visual acuity test. Both of these methods have their advantages and limitations. The electrophysiology test (visual evoked potential) is invasive, and animals need recovery time after being implanted with electrodes. Also,the electrophysiological visual acuity test only reflects the electrical activity of a single cell or nuclei, it does not reflect the overall visual function of the animal. The widely used behavioral visual acuity test is able to compensate for these limitations. This paper gives a brief overview of the methods of the behavioral visual acuity test for rodents (rats, mice, guinea pigs,etc.).
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· AIM:To compare the effect of fixation and select the optimal fixation solution and condition for PAS staining of guinea pig eyes among three different fixation solution:4% paraformaldehyde solution,4% glutaraldehyde solution and Davidson solution.· METHODS:Totally 30 healthy guinea pigs were divided into 6 groups:Group Ⅰ-Ⅱ were fixed in 4% paraformaldehyde solution and 4% glutaraldehyde solution for 24h,respectively;Group Ⅲ-Ⅴ were fixed in Davidson solution for 3,6 and 24h,respectively;and Group Ⅵ were fixed in Davidson solution for 3h and then transferred into 10% neutral formaldehyde solution for 48h.All groups were sectioned by routine section method and undergone PAS staining,and then observed by light microscope.· RESULTS:It was found that the group which was fixed by Davidson solution for 3h,remained the most complete structure for PAS staining (Group Ⅲ).While the effect of fixation for the group which was transferred into 10% neutral formaldehyde solution for preserving for 48h after fixing in Davidson solution for 3h was also acceptable for PAS staining (Group Ⅵ).The retinal cells remained clear and in order for both groups which was mentioned above.· CONCLUSION:The best fixation condition for PAS staining for eyes of guinea pigs is fixation in Davidson solution for 3h among these fixation conditions,while it is also suitable to transfer the eyes into neutral formaldehyde solution after fixing in Davidson solution for 3h for preserving for long periods,which is not severely reduced the effect of fixation for PAS staining.
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·AIM: To observe the efficacy and safety of a new posterior chamber intraocular lens ( ICMO V4c ) implantation in patients with high myopia and low astigmatism and to provide reference for the clinical diagnosis and treatment of these patients. ·METHODS: The study was to analyze the clinical data of patients with high myopia who underwent ICMO V4c implantation with low degree of astigmatism in our hospital from January 2015 to December 2016. The patients were followed up for 1, 3 and 6mo after operation. The items we analyzed including preoperative and postoperative uncorrected visual acuity ( UCVA) and best corrected visual acuity ( BCVA), diopter, spherical and cylinder equivalent, and manifest refraction spherical equivalent ( MRSE ), intraocular pressure ( IOP ), endothelial cell counting, anterior chamber depth (ACD), arch height and postoperative complications. ·RESULTS: Postoperative UCVA and BCVA improved compared with preoperative, and the difference between preoperative and postoperative was statistically significant (P<0. 05). All spherical and cylinder equivalent, and MRSE were reduced after surgery ( t = 38. 510, 20. 100, 34. 300; P<0. 01). The mean intraocular pressure increased at 1 and 3mo after operation ( t= 3. 998, 2. 837, all P<0. 05), but it returned to normal at 6mo ( t=0. 383, P>0. 05). The corneal endothelium counts in the patients at 3mo and 6mo after surgery were lower than those before surgery (t= 2. 119, 2. 411; all P<0. 05). The depth of anterior chamber was reduced to a certain extent within 6mo after operation, and recovered after surgery at 6mo after surgery, but it was still lower than the preoperative level (t=5. 850, 5. 260, 2. 556; all P<0. 05). The overall level of ocular arch after operation was stable and no significant fluctuations were observed. The incidence of postoperative complications in the surgical eye was low and significantly improved after active treatment. ·CONCLUSION: The new posterior chamber intraocular lens implantation in the treatment of high myopia with low astigmatism reaches satisfaction with less complications, is a good supplement to corneal refractive surgery.
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Objective To investigate the effects of Longdan Xiegan Decoction on the expression of Notch signaling-related genes in rats with experimental autoimmune uveitis (EAU).Methods Totally 24 Lewis rats were randomly divided into normal control group,EAU model group and Longdan Xiegan Decoction treatment group.A rat model of EAU was established in the EAU model group and Longdan Xiegan Decoction treatment group,followed by intragastrical administration of Longdan Xiegan Decoction in the latter group after successful modeling.On day 12 after different treatments,the spleen and lymph nodes were isolated from rats of all the three groups for the collection of CD4 + T ceils,and the levels of both Th17 and Treg cells were analyzed by flow cytometry to calculate the ratio of Th17 to Treg cells.Meanwhile the levels of Notch1,Notch2,Notch3,Notch4 mRNAs expression were monitored by qRT-PCR.Moreover,the expressions of Notch signaling-related proteins were further detected using ELISA technique.Results Flow cytometry showed increased Th17 cell level but decreased Treg cell level in the EAU model group when compared with the normal control group.However,after treatment with Longdan Xiegan Decoction,the Th17 level was decreased,whereas the Treg level was increased,and the ratio of Th17 to Treg gradually restored to equilibrium when compared to EAU model group.qRT-PCR results showed that the expression levels of Notch1,Notch2 and Notch4 mRNAs in Longdan Xiegan Decoction treatment group were significantly higher than those in the normal control group (all P < 0.01),but lower than those in EAU model group on day 12 after treatment (all P < 0.01),without Notch 3 expression in the spleen and lymph nodes.In addition,Notch 1,Notch 2 and Notch 4 protein levels of the spleen and lymph nodes in Longdan Xiegan Decoction treatment group showed the similar tendency as compared to those of Notch 1,Notch 2 and Notch 4 mRNAs (all P < 0.01),and the expression of Notch 3 protein was still not observed.Conclusion Longdan Xiegan Decoction can effectively relieve the imbalance of Th17/Treg cells in EAU rats,and its mechanism may involve the differentiation of naive CD4+ T cells into Th17 and Treg cells,which is regulated by Notch signaling.
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Objective To evaluate the effectiveness and safety of modified scleral tunnel for the prevention of Ahmed valve tube exposure in Ahmed valve implantation for refractory glaucoma.Methods A retrospective study was conducted in 72 patients with Ahmed glaucoma valve implantation for refractory glaucoma.All the patients were divided into unmodified and modified scleral tunnel group.In the unmodified scleral tunnel group,38 patients (38 eyes) underwent traditional scleral tunnel in Ahmed valve implantation,while in the modified scleral tunnel group,34 patients (34 eyes) received modified scleral tunnel in the procedures.The changes in intraocular pressure and visual acuity after operation were observed,and the incidence of drainage tube exposure and other complications were compared between the two methods after Ahmed valve implantation.The patients were followed up for 6 to 36 months with an average of 18 months.Results The intraocular pressure was (16.30 ±5.73) mmHg (1kPa =7.5 mmHg) in the unmodified scleral tunnel group and (15.80 ± 6.12)mmHg in the modified scleral tunnel group,with no significant difference between the two groups (P > 0.05),but there was significant difference with the preoperative data (48.4 ± 5.79) mmHg (P < 0.01).During the follow-up,conjunctival tube exposure was seen in 3 eyes (7.9%) in the unmodified scleral tunnel group,whereas there was no tube exposure in the modified scleral tunnel group.Conclusion The modified scleral tunnel is capable to prevent conjunctival tube exposure in patients with refractory glaucoma,which is more efficient and safe than traditional scleral tunnel in Ahmed glaucoma valve implantation.
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Objective To investigate the regulatory roles of rno-miR-30b-5p in the expression of Atg5,Atg12 and Becn1 autophagy genes and their expressions in rats with experimental autoimmune uveitis (EAU).Methods Application of dual luciferase report system was conducted to detect the regulatory roles of rno-miR-30b-5p in Atg5,Atg12 and Becn1 gene expression.Lewis rats were randomly divided into control group and EAU group,with 6 rats in each group.Next,rats in EAU group were immunized to establish EAU model.After treatments,Genesis-D fundus camera was used to observe the fundus inflammation every day.After immunization for 12 days,the pathological features of rat ciliary body and retina were detected,and meanwhile,the spleen and lymph nodes in both groups were isolated to detect the expressions of rno-miR-30b-5p,Atg5,Atg12 and Becn1 genes by quantitative PCR (Q-PCR);the levels of autophagy related proteins were determined by ELISA.Results Dual luciferase report gene expression assay confirmed that Atg5,Atg12 and Becn1 were target genes of rno-miR-30b-5p.Twelve days after immunization,compared with the control group,rats in the EAU group had severe iris adhesions and severe blood vessel swelling,and pathological examination revealed massive infiltration of inflammatory cells in the ciliary body and retina.Furthermore,rno-miR-30b-5p mRNA was 0.46 ±0.01,0.29 ±0.17in the spleen and lymph nodes in EAU group,respectively,which was down-regulated when compared with the control group (P <0.01);whereas the expressions of Atg5,Atg12 and Becn1 were significantly upregulated,and the differences were statistically significant (all P < 0.05).ELISA results showed that Atg5,Atg 12 and Becn 1 protein expression levels in the spleen and lymph nodes in EAU rats were significantly higher than those in the control group (all P < 0.05).Conclusion rno-miR-30b-5p can regulate the expressions of Atg5,Atg12 and Becn1 autophagy-related genes.The down-regulation of rno-miR-30b-5p expression in the spleen and lymph nodes in EAU rats can significantly up-regulate the expressions of Atg5,Atg12 and Becn1 genes,thereby regulating the pathogenesis of uveitis.
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It has been received wide attention for the increasing prevalence of myopia worldwide in recent years.The myopic fundus lesions,which may be associated with the variation level of the thickness of retinal and choroidal membrane in the early stage of myopia,remained the main cause of vision impairment and reduce the quality of life.However,the changes in retinal and choroidal thickness in the progress of myopia and the associated factors are not clear.This article gives a brief review the change in the thickness of retinal and choroidal thickness in progress of myopia and its associated factors,which can provide support for clinical diagnosis,prevention and monitoring of macula/choroidal associated diseases.
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Objective To investigate the effects of amphiregulin antibody on the axial length,diopter and posterior sclera thickness in eyes of bilateral lens-induced myopic guinea pigs.Methods A total of 60 healthy three-colored short-hair guinea pigs were randomly divided into 5 groups:myopic model group,low dose group,medium dose group,high dose group and normal control group,12 rats in each group,and both eyes of the guinea pigs in the former four groups were induced by-10 D lens,while the normal group did not make any treatment.After wearing of goggles for 2 weeks,the right eyes of guinea pigs were intravitreally administrated with 5 μg,10 μg,20 μg anphiregulin antibody for the low,medium and high dose group,respectively.At the same time,the left eyes of these groups were given the same dose of Ringer' s buffer (buffer group).Continuous wearing of the lenses before and after injection of antibody was allowed.Diopter and axial length of guinea pigs were measured before wearing of the lenses,2 weeks after wearing lenses and 5 weeks after the experiment (after 3-times injections of antibody).Moreover,the thicknesses of retina,choroid and sclera in the posterior pole were detected by PAS staining.Finally,the expression of amphiregulin and EGFR mRNA and protein was detected by real time quantitative PCR and immunofluorescence.Results After 2 weeks of lens induction,the axial length of the myopic model group increased,but the diopter decreased when compared with the normal control group,and the differences were statistically significant (both P < 0.05).Compared with the buffer group,after intravitreal injection of amphiregulin antibody,the axial length in the low,medium,and high dose groups decreased,while the diopter increased,and the scleral thickness at the posterior pole increased significantly in a dose-dependent manner,with statistically significant differences (all P < 0.05).However,there was no significant change in retinal thickness at the posterior pole of all groups.Real-time quantitative PCR and immunofluorescence showed the expression of amphiregulin mRNA and protein in the retina was upregulated the myopic model group and buffer group,but downregulated in the high,medium,and low dose groups.Furthermore,when compared with the buffer group,the expression level of epidermal growth factor receptor in the retina of the low dose group was decreased (t =2.606,P =0.022),but there was no significant difference in the other groups.Conclusion After injection of amphiregulin antibody into the eyes of bilateral lens-induced myopic guinea pigs,the diopter increases,but the axial length is significantly shortened,and the posterior sclera is thickened,which may involve a decrease in the expression of amphiregulin.
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In view of the anatomical and physiological barrier of the ocular surface and the intraocular structure, the conventional ophthalmic agents cannot efficiently reach the lesion site.Currently, the different types of nanomaterials possess great advantages in delivering drugs due to their characteristics of small size, easy preparation, degradability, strong targeting and less irritation to biological tissue.As drug delivery carriers, nanomaterials have been widely used in ocular drug delivery so as to treat different types of eye diseases.In this paper, the applications of nanomaterials as drug delivery carriers in treating eye diseases are briefly reviewed.
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<p><b>OBJECTIVE</b>To study the influence of Yanyankang powder on Th1/Th2 in rats with experimental autoimmune uveitis (EAU).</p><p><b>METHODS</b>The EAU models were induced in Lewis rats by immunization with interphotoreceptor retinoid binding protein (IRBP) 1177-1191 in complete Freund's adjuvant. The rats were randomly divided into 3 groups: a model control group, a Yanyankang group, and a prednisone group, 9 rats in each group. The model control group was intervened with saline solution by gavage. The Yanyankang group was intervened with Yanyankang powder 4 g/(kg day) by gavage. The prednisone group were intervened with prednisone acetate tablets 5 mg/(kg d) by gavage. All groups were intervened after immunization once every 2 days for 18 days and monitored by slit-lamp biomicroscopy daily until day 18. The levels of gamma interferon (INF-γ) and interleukin-10 (IL-10) in the supernatants of T cells were determined by enzyme-linked immunosorbent assay. Polymerase chain reaction (PCR) technology was used for measuring Th1 and Th2 related cytokine mRNA expressions.</p><p><b>RESULTS</b>Slighter intraocular inflammation was found in the Yanyankang group and the prednisone group than the control group. The levels of the IFN-γ and IL-10 in the supernatants of the spleen lymph node cells were 382.33±6.30, 155.87±4.46 μg/L in the Yanyankang group and 270.93±7.76, 265.32±11.88 μg/L in the prednisone group. Both had significant differences compared with the control group (941.53±8.59, 20.67±4.65 μg/L; =0.01). The PCR results showed the same tendency.</p><p><b>CONCLUSION</b>Yanyankang powder showed favorable effects in the rats with EAU by influencing the function of Th1 and Th2 cells.</p>
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Animales , Femenino , Medicamentos Herbarios Chinos , Farmacología , Usos Terapéuticos , Ojo , Patología , Inmunización , Inflamación , Patología , Interferón gamma , Genética , Metabolismo , Interleucina-10 , Genética , Metabolismo , Ganglios Linfáticos , Metabolismo , Polvos , ARN Mensajero , Genética , Metabolismo , Ratas Endogámicas Lew , Bazo , Metabolismo , Células TH1 , Alergia e Inmunología , Células Th2 , Alergia e Inmunología , Uveítis , Quimioterapia , Genética , Alergia e InmunologíaRESUMEN
Myopia is one of the most popular eye diseases all over the world. The development of the current understanding of its mechanism is still limited. Many studies indicated that the growth factors closely related to eye development and myopia. Some growth factors with biological activity, such as transforming growth factor ( TGF ), fibroblast growth factor ( FGF ) and epidermal growth factor ( EGF ), have an impact on scleral thickness variation, the regulation of the development of myopia and so on, which plays a non-negligible role in the pathogenesis of myopia. In this paper, the function of various growth factors in myopia will be reviewed.
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Background C57BL/6(B6) is a kind of routine mouse specie used in experimental autoimmune uveitis (EAU) research.Previous studies showed that the pathogenesis of uveitis related to inflammatory cytokines secreted by different helper T(Th) cells.However,the interaction of different Th cells in EAU is unclear.Objective This study was to investigate the dynamic changes of inflammatory factors in the spleen and serum after immunization in EAU mice.Methods Forty-four SPF B6 mice were immunized by injection of interphotoreceptor retinoid-binding protein (IRBP) and complete Freund adjuvant (CFA) emulsion via caudal vein and footpad.Indirect ophthalmoscope was used to examine the eyes 3 times per week and the inflammatory response was scored based on Thurau's criteria.In the thirty day after injection,20 model eyes were extracted and the sections of eye tissue were prepared for histopathological examination.The spleens of model mice were enucleated before injection and 2,5,10,15,20,25,30 days after injection,and reverse transcriptase PCR (RT-PCR) was used to detect the contents of interleukin-17 (IL-17) mRNA,interferon-γ (IFN-γ) mRNA,tumor necrosis factor-α (TNF-α) mRNA and IL-10 mRNA,and the contents of IL-17,IFN-γ,TNF-α and IL-10 in model serum were assayed by ELISA in 24 model mice.The experimental protocol and use of the animals were approved by Ethic Committee for Care and Use of Laboratory Animals of Shandong University of Traditional Chinese Medicine.Results Mild inflammatory response was seen in 12 days under the indirect ophthalmoscope with the scores of 0.5.The inflammatory scores peaked in 13-15 days with the scores of 1.0 and alleviated after that with the inflammatory scores of 0.5 in 30 days after injection.The histopathological score was consistent with the clinical score in the models on the 30 days.The serum IL-17 content of model mice was (0.98±0.05) ng/L before injection and increased to (51.85 ±2.42) ng/L on the fifth day,and decreased to (4.01±0.06)ng/L on the fifteen day.But,the serum IL-17 level increased to (25.00±0.94)ng/L again on the 25th day,and then lowed to (6.01 ±0.21)ng/L 30 days after injection,showing a significant elevation in comparison with that of before injection (P=0.000).The serum IFN-γ content of the model mice was (1.02±0.09)ng/L before injection and increased to (50.54±0.48) ng/L on the fifth day,and (73.21±0.12) ng/L on the tenth day,and then it declined gradually until (5.15±0.18)ng/L in the 30th day,which was still higher than that of before injection (P=0.000).After injection of IRBP+CFA,the serum TNF-α level upregulated from the second day to fifth day with the peak values (134.25±0.59)ng/L,and declined to valley on 15th day.A repeat elevation of serum TNF-α level was found on the 20th day with the values (60.54±0.62)ng/L and followed by decrease till the 30th day,which was higher than that of before injection (P=0.660).Serum IL-10 was detectable in the tenth day and peaked on the 15th day.Then a slight decrease was seen till the 30th day,compared with before injection(P =0.000).The contents of IL-10 mRNA,IL-17 mRNA,TNF-α mRNA,IFN-γmRNA in mice spleens followed the same pattern with serum levels of their proteins.Conclusions IL-17,IFN-γ,TNF-α and IL-10 are key inflammatory factors of Th1,Th2 and Th17,they present with specific changes during EAU,it confirming that IFN-γ probably play a pathogenic role in EAU,IL-17 and TNF-α levels probably associated with the chronic and recurrent procedure of uveitis,IL-10 plays an inhibit role in EAU.
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Background The pathogenesiof age-related maculadegeneration (AMD) iassociated with the senility of human retinal pigmenepithelium (RPE) cells.Seeking drug to arresRPE cell senility iof significance fothe prevention and treatmenof AMD.Research showed thathe lycium barbarum polysaccharide (LBP) can delay senility,buitinfluence on RPE cell aging iunclear.Objective Thistudy wato discusthe protective effecand mechanism of LBP on RPE cell aging.MethodPorcine retinal neural epithelial layewaisolated,and photoreceptooutesegmen(POS) waextracted by density gradiencentrifugation and marked by FITC.The POwathen co-cultured with RPE cellin the medium containing 0.01,0.10 and 1.00 g/L LBP fo24 hours.The areof fluorescence,representing the amounof POphagocytosed by RPE cells,wameasured undethe fluorescenmicroscope to evaluate the influence of LBP on the phagocytifunction of RPE cells.The POS-induced RPE lipofuscin-uptake cell model waestablished by co-culturing human RPE cellwith porcine POfo3 weeks.The RPE-POco-culture cell model waincubated in medium containing 0.01,0.10 o1.00 g/L LBP,and the autofluorescence caused by lipofuscin up-taken into RPE cellwadetected with flow cytometry.cell counting kiwaused to assescell proliferation and viability (value) 24,48 and 72 hourafteculturing.ResultPorcine POpresented athin rodundethe lighmicroscope and appeared abilayedisc-like structureundethe transmission electron microscope,and itFITC-labeled yellow-green autofluorescence waobserved undethe fluorescenmicroscope.No POwaup-taken into the RPE cellin the normal control group,buthe areof POphagocytosed by RPE cellwagradually enlarged with increasing doseof LBP,showing significandifference among the group(F =21.425,P =0.006).Compared with the POcontrol group,the phagocytosed areincreased avariouconcentrationof LBP+POgroup(P<0.01).Flow cytometry showed thathe autofluorescence value in the POcontrol group wamore highethan thaof the normal control group.Athe LBP dose increased,the autofluorescence value in the RPE celldeclined gradually and iwaneathe normal value in the 1.00 g/L LBP+ POgroup.The rate of proliferation of the lipofuscin RPE cellvaried with the increase of doseof LBP with the maximal value in the normal RPE group and minimal value in the lipofuscin RPE group,and the rate of proliferation of the lipofuscin RPE cellascended with increasing doseof LBP until neathe normal value in the 1.00 g/L LBP + lipofuscin RPE cellgroup (P>0.05).ConclusionLBP enhance the anti-aging effecof human RPE cellby strengthening the phagocytiability to POand the ability to remove lipofuscin and by heightening the proliferation of human RPE cells.