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Objective:To investigate the effect and the neural mechanisms of Apelin-13 on the behavior changes of posttraumatic stress disorder (PTSD) model mice.Methods:Totally 32 SPF grade male C57BL/6J mice aged 6 weeks were divided into 4 groups randomly ( n=8 in each group): control group, model group, normal saline group and Apelin-13 group.The mice model of PTSD was established by single-prolonged stress (SPS) method. The mice in normal saline group and Apelin-13 group were respectively given lateral ventricular microinjection of 0.9% sodium chloride solution (2 μL) and Apelin-13 (1.5 μg/μL, 2 μL)after PTSD modeling. The behaviors of mice were evaluated by open field test, elevated plus maze test and Morris water maze test.The morphological structure and numerical changes of hippocampal neurons were observed by hematoxylin and eosin (HE) staining.The expression of phosphoinositide 3-kinase(PI3K), phosphorylated-PI3K(p-PI3K), protein kinase B(Akt), phosphorylated-Akt (p-Akt), forkhead box O3a (FoxO3a), phosphorylated-FoxO3a(p-FoxO3a), autophagy-related proteins including microtubule-associated protein 1 light chain 3(LC3) and sequestosome 1(p62) were detected by Western blot. SPSS 26.0 software was used for data analysis.The escape latency data of repeated learning training in Morris water maze was conducted by repetitive measurement ANOVA.The comparison of other data among multiple groups was conducted by one-way ANOVA and further pairwise comparisons were conducted by LSD test and Tamhane test. Result:(1) Open field test results showed statistically significant differences in the central area activity distance and residence time in central area among mice in the four groups ( F=15.37, 9.63, both P<0.05). The central area activity distance ((0.06±0.03) m) and residence time ((2.48±1.02) s) of the mice in model group were lower than those of the control group ((0.19±0.05) m, (15.00±8.91) s)(both P<0.05). And the central area activity distance((0.12±0.04)m)and the residence time((13.56±7.64)s)were higher than those of model group((0.06±0.03)m, (2.48±1.02)s)and normal saline group((0.06±0.02)m, (2.82±1.52)s)(all P<0.05). Elevated plus maze test results showed statistically significant differences in the numbers and time entering open arms among the four groups ( F=10.74, 19.12, both P<0.05). The numbers((4.50±2.51) times) and the time ((26.95±17.48) s) entering the open arm of mice in model group were both lower than those of the control group ((13.75±4.71) times, (103.75±42.43)s) and Apelin-13 group ((10.00±5.18) times, (55.98±19.49) s) (all P<0.05). Morris water maze test results showed that in the 4-day learning and training phase, the time and group interaction of escape latency was not significant among the four groups ( F=1.15, P=0.34), but time main effect and group main effect were significant ( F=131.65, 16.98, both P<0.05). On the 2nd to 4th day, mice in model group showed significantly increased escape latency than mice in control group and Apelin-13 group(both P<0.05). And the numbers crossing original platform and the time in the target quadrant of Apelin-13 group were both higher than those of model group and normal saline group (all P<0.05). (2) HE staining results showed that neurons in the hippocampal CA1 and CA3 area of mice in model group and normal saline group were swollen and arranged loosely.The hippocampal neurons in control group and Apelin-13 group were arranged neatly and densely. (3) Western blot results showed statistically significant differences in the protein expression of p-PI3K, p-Akt, p-FoxO3a, p62 and the ratio of LC3Ⅱ/LC3Ⅰ among the four groups ( F=21.37, 37.35, 20.71, 13.26, 37.65, all P<0.05). The protein expression of p-PI3K, p-Akt, p-FoxO3a and p62 in Apelin-13 group((0.92±0.07), (0.90±0.09), (0.89±0.13), (1.03±0.08)) were higher than those in model group((0.59±0.04), (0.50±0.07), (0.49±0.11), (0.68±0.04)) and normal saline group((0.61±0.06), (0.50±0.08), (0.53±0.11), (0.70±0.05))(all P<0.05), and the ratio of LC3Ⅱ/LC3Ⅰ in Apelin-13 group(0.60±0.06) was lower than those in model group(0.92±0.10) and normal saline group(0.99±0.05) (both P<0.05). Conclusion:Apelin-13 can alleviate the anxiety-like behavior and impaired spatial learning and memory in PTSD model mice. The mechanism may be related to the up-regulation of PI3K/Akt/FoxO3a autophagy pathway.
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Objective To investigate the risk factors of postoperative vitreous hemorrhage after minimal vitrectomy without endotamponade for proliferative diabetic retinopathy (PDR).Methods From June 2015 to June 2017,103 eyes of 103 patients with PDR diagnosed and underwent minimalvitrectomy in Henan Provincial People's Hospital were enrolled in the study.There were 58 males and 45 females,with the average age of 58.37± 10.14 years and diabetes duration of 8.7± 7.2 years.Baseline systemic parameters including sex,age,diabetes duration,hypertension,HbA1c,creatinine,whether received anticoagulants,ocular parameters including whether combined with vitreous hemorrhage,whether finished panretinal photocoagulation (PRP),whether received treatment of anti-VEGF,whether combined with iris neovascularization (NVI),lens status preoperatively,whether hypotony postoperatively and intraoperative parameters including whether disc neovascularization (NVD) bleeding,whether fibrovascular membrane (FVM) residual,laser points,whether combined with cataract phacoemulsification were identified by multivariate logistic regression analysis.Results Twenty-nine of 103 eyes (28.15%) developed PVH in 1 day to 6 months after surgery,with self absorption of 18 eyes and reoperation of 11 eyes.Univariate analysis showed there were significant differences in age (t=2.124,P=0.036),anti-VEGF(x2=7.105,P=0.008),NVD bleeding (x2=10.158,P=0.001) and FVM residual(x2=8.445,P=0.004) between patients with and without postoperative vitreous hemorrhage.Sex (x2=0.021,P=0.884),diabetes duration (t=0.87,P=0.386),hypertension (x2=2.004,P=0.157),HbA1c (t=1.211,P=0.229),creatinine (t=0.851,P=0.397),preoperative oral anticoagulants (x2=0.985,P=0.321),preoperative vitreous hemorrhage (x2=0.369,P=0.544),PRP (X2=1.122,P=0.727),NVI (x2=2.635,P=0.105),lens status (x2=0.172,P=0.679),hypotony postoperatively (x2=1.503,P=0.220),laser points (x2=1.391,P=0.238) and combined phacoemulsification surgery (x2=0.458,P=0.499) were not associated with PVH.Multivariate logistic regression analysis revealed the more PVH appeared in younger (OR=1.065,P=0.009) and NVD bleeding (OR=6.048,P=0.001) patients.Conclusion Younger age and NVD bleeding are the important risk factors for PVH after minimal vitrectomy without endotamponade in PDR.
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Objective To observe the occurrence and evolution of persistent submacular fluid (SMF) after scleral buckling surgery (SB) in rhegmatogenous retinal detachment,and then to study the related factors of persistent SMF and the effect of persistent SMF on visual outcome.Methods Ninety eyes of 89 patients with rhegmatogenous retinal detachment which had been performed SB were included in this study.Best corrected visual acuity (BCVA),intraocular pressure,slit-lamp microscopy,three mirror contact lens,indirect ophthalmoscopy and B-scan ultrasonography were measured for all patients.There were 21 eyes with atrophic holes while 42 eyes with horse-shoe tears,22 eyes with old retinal detachment while 68 new suffered eyes.Thirty-two eyes underwent scleral encircling surgery (SE) and 58 eyes underwent segmental scleral buckling surgery (SSB).The patients were divided into SMF group and non-SMF (NSMF) group according to the results of optical coherence tomography (OCT) at 1 month postoperatively.Thorough ophthalmologic examinations were performd at 1,3,6 and 12 months after surgery to the patients,further observations were continued to carry out unless the abnormality had resolved for at least 6 months.Results Patients who underwent SE (20 eyes,62.5 %) had a higher incidence of persistent SMF at 1 month after surgery than those who underwent SSB (23 eyes,39.7 %),the difference was significant (x2 =5.024,P< 0.05).Persistent SMF was more frequent in eyes with atrophic holes (66.7%) than that with horseshoe tears (38.1%),the difference was significant (x2 =4.582,P<0.05).Persistent SMF was found in 72.7% old retinal detachment eyes and in 39.7% new suffered eyes,showed a striking differences (x2=7.264,P<0.01).There was no significant difference in BCVA among SE and SSB groups at every time point (t=0.659,0.699,1.108,1.037,1.902; P>0.05).The SMF group have a similar BCVA with NSMF group 1 and 3 months after surgery (t=1.812,1.957; P>0.05),whereas the SMF group showed worse BCVA than NSMF group from since 6 months after surgery (t=2.324,2.147,2.184; P<0.05).Conclusions Persistent SMF is more frequent after SE than SSB,the type of retinal breaks and old retinal detachment may be the potential influencing factors.Persistent SMF after SB may affect the final visual outcome.
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Objective To investigate the main causes and risk factors of recurrent retinal detachment (RRD) after silicone oil removal (SOR) in eyes with complex retinal detachment.Methods It was a retrospective case series study.A total of 458 eyes of 455 consecutive patients who underwent pars plana vitrectomy with silicone oil tamponade were recruited in this study.All patients underwent vitrectomy operation.Additionally,they were given heavy water,membrane peeling,retinotomy or partial cutting,intraocular laser photocoagulation or frozen,gas-liquid exchange or direct oil exchange operation accordingly.Ninety-eight eyes with multiple holes,old retinal detachment,hyperplasia and serious traction lesions underwent scleral buckling surgery simultaneously.Intravitreal silicone oil was padded at the end of operation.Cutting,stripping or resection and 360° preventive laser photocoagulation were applied while the epiretinal membrane was found and need treatment during SOR.Holes or suspicious hiatus underwent intraocular laser photocoagulation or cryotherapy during the operation.One week after SOR and during follow-up,the visual acuity,intraocular pressure (IOP),slit lamp microscope,and ophthalmoscope examination were examined with the same technique and methods as preoperation.The eyes were divide into two groups based on the attachment status of retina after SOR,which were reattached group (419 eyes) and redetached group (39 eyes) respectively.The following data were recorded.,the age of patients,ocular axial length,logarithm of minimum angle of resolution (logMAR) best corrected visual acuity (BCVA) and IOP before vitrectomy operation and before and after SOR,the number of retinal breaks,the duration of silicone oil filling,the duration of follow-up,and the related factors during vitrectomy operation and SOR.The relation of age,sex,high myopia,the size and location of holes,aphakic eye,proliferative vitreoretinopathy (PVR) C3 level and above,previous history of failed retinal detachment operation,360° preventive laser photocoagulation,assistant scleral buckling surgery,SOR via corneal puncture to RRD after SOR were analyzed.Odds ratio (OR) and its 95% confidence interval (CI) were calculated for the age <40 years old and gender.High myopia,assistant scleral buckling surgery and SOR via corneal puncture were further analyzed by multiple regression equation.Results After SOR operation,the total average logMAR BCVA was 0.86 ± 0.63.The average logMAR BCVA was 0.82 ± 0.59 and 0.99 ± 0.70 respectively for the reattached and redetached groups,which was not statistically different (F=1.559,P>0.05).The number of high myopia eyes in the reattached and redetached groups were 116 and 22 eyes,respectively,accounted for 27.7 % and 56.4 %,and the difference was statistically significant (x2=13.984,P<0.01).Three eyes underwent vitrectomy with scleral buckling occured RRD,accounting for 3.1%; while 36 eyes underwent vitrectomy without scleral buckling occured RRD,accounting for 10.0%.The incidence of RRD between them was statistically significant (x2 =4.761,P<0.05).The incidence of RRD was not retated to the PVR levels before the operation,previous history of failed retinal detachment operation,aphakic eye and preventive laser photocoagulation (OR=1.626,1.699,1.986,0.709; 95% CI:0.836-3.162,0.832-3.658,0.921-4.279,0.268-1.875; P>0.05).RRD had a close relation with high myopia and assistant scleral buckling surgery (OR=3.380,0.284; 95%CI:1.733-6.595,0.086-0.944; P<0.05).The raise of risk derived from SOR via corneal puncture had no statistical significance (OR=2.119; 95%CI:1.043-4.306; P>0.05).The incidence of RRD after SOR was 8.5%; of which,35.9% originated from new breaks and 69.2% were related to new breaks,in contrast,only 5.1% originated from PVR but 51.3% were related to PVR.Conclusions High myopia is an independent prognostic risk factor of RRD after SOR.Combined scleral buckling surgery is a protective factor of RRD after SOR.To the well reattached eyes before SOR,the new breaks seems to be the main cause of RRD,wheras PVR was probably a secondary phenomenon.
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<p><b>OBJECTIVE</b>To establish a animal model of hepatic steatosis induced by chronic viral hepatitis in C(57)BL/6 mice.</p><p><b>METHODS</b>C(57)BL/6 mice were randomly assigned to control group, high-fat diet group, mouse hepatitis virus strain A59 (MHV-A59) virus infection group, and high-fat diet plus virus infection group. At 13 weeks of the experiment, serum samples were collected to detect MHV antibodies and transaminase and lipid levels. The hepatic pathologies of the mice were examined with Oil red O staining of the frozen sections the and HE staining of paraffin-embedded sections.</p><p><b>RESULTS</b>The mice in the two virus infection groups showed strong positivity of MHV antibodies in the serum. Compared with the control group, the mice in high-fat diet group and the two virus infection groups had significantly increased AST and ALT levels with also elevated TC and LDL-C levels. The two virus infection groups both exhibited obvious pathologies in the liver characteristic of chronic viral hepatitis with increased lipid accumulation in the hepatocytes.</p><p><b>CONCLUSION</b>We have successfully established a mouse model of hepatic steatosis induced by chronic viral hepatitis, which provides the basis for further study of the disease mechanism.</p>
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Animales , Ratones , Anticuerpos Antivirales , Sangre , Enfermedad Crónica , Dieta Alta en Grasa , Modelos Animales de Enfermedad , Hígado Graso , Virología , Hepatitis Crónica , Virología , Ratones Endogámicos C57BL , Virus de la Hepatitis MurinaRESUMEN
The effect of hydroxyprolpyl methylcelulose (HPMC) injected into anterior chamber on filtering angle tissues was studied. Twenty-four rabbits were randomly divided into two groups: the experimental group included 40 eyes receiving HPMC which was injected into anterior chamber and the control group included 8 eyes (one served as normal control and the remaining were injected with balanced salt solution). According to the intaocular pressure (IOP) was normal or elevated ,the experimental group was further divided into group A and B. The tissul specimen of filtering angle were collected to perform pathological examination at 5 days, 3 weeks ,and 10 weeks after injection.During the early period, in groupA,the trabecular meshwork dilated slightly, the vacuoles increased in endothelial cells of inner wall of Schlemm's canal. In group B, the space of trabeculum broadened, there were accumulation of collagen and intercellular fibrosis, the vacuoles significantly increased in endothelial cells of inner wall of Schlemm's canal. But the change mentioned above all recovered to normal appearance on the later stage of long term following observationHPMC injected into anterior chamber may cause temporary elevation of IOP and pathological change of filtering angle, but the change was reversible. HPMC is not toxic or destructive to the tissues of anterior chamb angle.
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Objective To investigate the effects of intravitreal injection of matrix metalloproteinase-3 (MMP-3) on the vitreoretinal adhesion and the vitreous gelatin. Methods Twenty-four pigmented rabbits were randomly divided into 3 experimental groups (group A, B, and C)and one control group with 6 rabbits (12 eyes) in each. Different concentrations of 0.1 ml MMP-3 (5,10, 20 ng in group A, B, and C, respectively) and equivalent dose of balanced salt solution were intravitreally injected to the rabbits, respectively. Clinical examinations (such as gross observation, slit-lamp biomicroscopy, indirect fundus ophthalmoscopy ), electroretinography (ERG) and fundus fluorecein angiography (FFA) were taken before and after injection. Results One week after injection, posterior vitreous detachment (PVD) and focal vitreous liquefaction were recognized clinically for the first time in 1 eye in group B. By the end of this study, clinically detected PVD developed in 1 eye in group A, 3 eyes in group B, but the synchisis developed slowly, and no liquefaction or PVD occurred in control group. As for the histological examination, partial PVD was observed in 1 eye in group A and 3 eyes in group B 60 minutes after injection. All of the eyes in group A and B showed partial PVD 1 week after injection, and the area of PVD enlarged in contrast with before. Complete PVD were recognized in 1 eye in group A and 3 eyes in group B 15 weeks after injection, and the cleavage was narrow and limited. In group C, inflammatory cell infiltration in the inner layer of retina, destruction of retinal structure, and fluorescein leakage at late phase was found in the early period after injection. Conclusions MMP-3 is effective in disrupting the adhesion of the posterior hyaloid to the inner limiting membrane leading to PVD, and helpful to some extent in producing vitreous liquefaction. The dose of 10 ng MMP-3 is safe and effective for the rabbits eyes, which may be used as an promising assistant of vitreous surgery.