Objective:To explore the establishment and evaluation method of recurrent infection model of herpes simplex keratitis (HSK).Methods:To determine the optimal duration of the ultraviolet light irradiation, 12 healthy BALB/c mice were randomized into 3 minutes group, 2 minutes and 45 seconds group, and 2 minutes and 30 seconds group based on the duration of the ultraviolet light irradiation according to random number table method, with 4 mice in each group.Another 72 healthy BALB/c mice were randomized into blank control group, model group and recurrence group according to random number table method, with 24 mice in each group.The 72 mice were scratched a # symbol on their right corneas with scalpel.Then the eyes in the blank control group were treated with 5 μl of normal saline solution, while the eyes in the model group and recurrence group were treated with 5 μl of herpes simplex virus I (HSV-1) suspension.All the 72 mice were not intraperitoneally injected with HSV-1 xenogeneic serum antibody.Five weeks after the initial infection, the mice in the recurrence group were irradiated at ultraviolet B-302 nm for the optimal duration.The feasibility of establishing HSK recurrence model induced by this method was evaluated by the score of ocular surface symptoms, and the cytopathic effect (CPE) lesion analysis of Vero cells cultured in corneal wiping fluid from the mice in the model group and recurrence group.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology (ARVO). The study protocol was approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine (No.PZSHUTCM190222039).Results:The optimal duration of the ultraviolet light irradiation was 2 minutes and 45 seconds.Within one week, HSK syndromes appeared in the mice injected with HSV-1 virus in the model group and recurrence group, and then gradually disappeared after one week.Before inducing recurrence, slit lamp examination was performed, and there was no spontaneous recurrence of HSK in the model group and recurrence group.The mice in the recurrence group relapsed within one week after ultraviolet light irradiation.The symptom scores of ocular surface lesions in the blank control group, model group and recurrence group were 0.333±0.471, 1.500±0.764 and 2.667±0.943 at one day after ultraviolet light irradiation, 0.000±0.000, 0.833±0.373 and 5.167±2.267 at three days after ultraviolet light irradiation, and 0.167±0.373, 1.000±0.577 and 3.000±1.155 at seven days after ultraviolet light irradiation, respectively.The symptom score of ocular surface lesions was higher in the recurrence group than the blank group and the model group at three days after ultraviolet light irradiation, and the differences were statistically significant (both at P<0.05). Morphological changes such as floating and gathering were found in the Vero cells after being cultured in corneal wiping fluid.The positive rate of CPE lesion was 71% (17/24) in the recurrence group, which was significantly higher than 8% (2/24) in the model group, and the difference was statistically significant ( P<0.01). Combining the two indicators, the success rate of recurrence in established models could reach 71%. Conclusions:Ultraviolet light irradiation can successfully induce the recurrence of viral keratitis in HSK mice without injection of neutralizing serum antibody.

Objective To investigate the roles of somatostatin(SS)positive intemeurons in the development and compensation of temporal lobe epilepsy.Methods Piloearpine-induced epilepsy rat model was established.Immunohistochemistry method was used to detect number changes and axonal sprouting of SS positive intemeurons in different domains of the hippocampus at difierent time points.Degeneration of SS positive interneurons and their neurophils were detected by the double immunofluorescence staining with SS and Fluoro-Jade B(FJB)at 7 and 60 days after status epilepticus (SE).Results In the exoerimental rat group,the number of SS positive neurons decreased in each hippocampal domain,and it reached the lowest at 7 days post-SE(There were 11.1±3.3 in hilus,2.8±0.9 in CA1region and 1.8±0.7 in CA1region,t=13.519,9.644 and 8.808,all P<0.01).In chronic phase,the number of SS neurons gradually recovered,and exceeded the control group in CA1 area at 60 days post-SE(12.8±1.5 vs 8.8±1.3,t=-4.506,P<0.01),however,the number of SS neurons in the hilus(25.5±4.6)and CA1 area(4.8±0.8)remained significantly less than normal levels(t value were 4.691 and 3.953.both P<0.01).Increased SS positive fibers were found in the lacunosum-molecular (1m)layer and outer molecular layer of dentate gyrus after 30 days post-SE,and numerous SS positive fibers were seen threnghout the layers of area CA1 at 60 days post-SE.Double immunofluuorescence revealed that a few SS positive interneurons and fibers were also labeled by FJB in area CA1 at 7 days post-SE and in CA domain/hilus at 60 days post-SE.Conclusions SS intemeurons loss plays an important role in the development of temporal lobe epilepsy.The loss is partially caIlsed by the degeneration and death of neurons;SS positive neurophils increase within area CA1 in chronic phase may play a significant role in the generation and compensation of temporal lobe epilepsy.

Objective To investigate the expression of XIAP, Smac, HtrA2 and XAF1 in the hippocampus following SE in rats and to explore the pathophysiological mechanisms of expression of XIAP and its negative regulators after SE. Methods The lithium-pilocapine model of status epilepticus was established in SD rat. XIAP, Smac, HtrA2, XAF1 and activated caspase-3 protein were examined using immunohistochemistry. Western blot was used to detect the protein levels of XIAP, Smac, HtrA2 and activated easpase-3. Results XIAP immunoreactivity diffusely distributed within the neuron after SE. Compared with the control group, the expression of CA3 XIAP protein in the SE group was increased gradually since 2 hours (0.5503±0.0172 vs 0.1507±0.0165, t=115.87, P<0.01), peaking at 8 hours (0.6221±0.0238 vs 0.1507±0.0165, t=136.69, P<0.01). The expression of CA3 Smac, HtrA2, XAF1 and activated caspase-3 protein were increased generally following SE. Western blot analysis showed a significant increase in Stoat, HtrA2, activated caspase-3 protein levels from 2 to 72 hours following SE, but no significant differences were seen in XIAP protein levels between the control group and the SE group. Conclusions The XIAP, Smac, HtrA2 and XAF1 are involved in the regulation of neuronal apoptosis and implicated in pathophysiological mechanisms of neuronal damage after SE.

Objective To identify the differences of 3D geometry of internal carotid artery (ICA)siphon between the controls and patients with ICA siphon stenosis. Methods The clinical and imaging data of the inpatients underwent carotid artery MRA in the past three years were collected. All patients were divided into the control group ( 17 males and 14 females with mean age of 67. 5 years) and ICA siphon stenosis group (20 males and 9 females with mean age of 58.6 years). There were 5 smokers and 9 smokers in two groups, respectively. The atherosclerotic predisposing factors were compared between the two groups using chi-square test and paired t-test. In order to extract the 3D geometry of ICA siphon, the MRA data were transferred to PC and processed with the software of Mimics. The average curvature radius (ACR) was calculated and paired t-test was applied to determine the bilateral differences in the controls. According to the stenotic site of ICA siphon, ICA siphon stenosis group was divided into C2 segment stenosis group and C4 segment stenosis group. The differences of ACR among the control group, C2 segment stenosis group and C4 segment stenosis group were compared. In addition, the values of ACR in the stenotic and normal sides were compared with paired t-test in patients with unilateral C2 segment stenosis. Results No significant differences were found in gender and smoker between the control group and the ICA siphon stenosis group ( χ2 = 1.63, P ＞ 0. 05; χ2 = 1.86, P ＞ 0. 05 ). The systolic blood pressure was ( 146. 6 ± 21.3 ) mm Hg ( 1 mm Hg =0. 133 kPa) and ( 140. 3 ± 17. 3) mm Hg respectively in the ICA siphon stenosis group and the control group. The serum glucose level was ( 5.94 ± 1.89 ) mmol/L and ( 6. 79 ± 3.57 ) mmol/L respectively in two groups. The serum cholesterol level and triglyceride level were (4. 57 ± 0. 87 ) mmol/L,( 1.34 ± 0. 63 ) mmol/L and (4. 75 ± 1.70) mmol/L, ( 1.54 ± 0. 72) mmol/L respectively in two groups.There were no differences in the atherosclerotic predisposing factors between two groups (t = 1.24, 1.16,0. 71 and 1.16 respectively,P＞ 0. 05). In the control group, the ACRs on the left and right were (3. 82 ±0. 69) mm and (4. 08 ± 1.04) mm respectively and no difference was found ( t = - 1.44, P ＞ 0. 05 ).Kruskal-Wallis test revealed that there were significant differences among the controls, C2 segment stenosis group and C4 segment stenosis group ( χ2 = 6. 67, P ＜ 0. 05 ). The ACR was significantly different only between the controls and C2 segment stenosis group ( t = 2. 63, P ＜ 0. 05 ). The stenotic side had a tendency to have less ACR than the normal side (t =2. 09, P =0. 05) in patients with unilateral C2 segment stenosis. Conclusion Patients with smaller ACR are more susceptible to have the stenosis of C2 segment in ICA siphon, and 3D geometry may be a possible reason for the development of the ICA stenosis.

Objective To quantitatively assess the cardiopulmonary exercise function of spinal cord injury (SCI) patients and observe the effect of aerobic exercise on cardiopulmonary function, motor function and activities of daily living. Methods From December, 2014 to June, 2016, 34 incomplete SCI patients (ASIA C and D) and 23 healthy controls received cardiopulmonary exercise test (CPET). SCI pa-tients were randomly divided into conventional rehabilitation group (n=17) and aerobic exercise group (n=17). The aerobic exercise group received aerobic exercise for four weeks. They were assessed with CPET, motor and sensory function, walking index for spinal cord injury II (WISCI II) and spinal cord independence measure (SCIM) before and four weeks after training. Results Oxygen uptake (VO2)peak, anaerobic threshold (AT), metabolic equivalent of energy (METpeak), VO2/heart rate (HR)peak, respiratory exchange rate (RER)peak, minute ventilation (VE)peak, work rate (WR)peak and systolic blood pressure (SBP)peak were lower in the patients than in the controls (t>2.714, P<0.05). VO2peak、AT、METpeak、VO2/HRpeak、WRpeak increased in the aerobic exercise group after training (t>2.431, P<0.05). METpeak and WRpeak improved in the conventional rehabilitation group after training (t>3.282, P<0.01). The scores of motor in ASIA and SCIM improved in both groups after training (t>2.985, P<0.05). Conclusion The cardiopulmonary function decreased in incomplete SCI patients, which could be improved by moderate intensity aerobic exercise.

Objective To compare the fluorescence intensity and duration of qdots streptavidin conjugate (QDs-SA) with Cy3 as the molecular probe of β amyloid precursor protein (APP), and to provide evidence for early molecular imaging and diagnosis of Alzheimer's dissease (AD). Methods With the help of laser scanning confocal microscope and flow cytometry, the flurescence probe based on the QDs-SA was used to detect APP in HEK293 cells stably transfected pcDNA3.1/APP, and to compare with conventional fluroimmunoassay Cy3. Results The immunofluorescence staining detection indicated APP expression was mainly located in the plasma membrane. The mean fluorescence intensity of QDs-SA (34.2336±4.2455) was greater than that of Cy3 (21.6023±3.0102)under the confocal fluorescence microscope (P<0.05). After persistent exciting for 12 min, the fluorescence intensity of APP stained by QDs-SA decreased by 27.87%. The other stained by Cy3 decreased by 79.60%. The positive rate of APP staining had no significant difference between the QDs-SA(54.4700±3.4433)% and Cy3 (54.3800±8.5229)% by flow cytometry, but the mean fluorescence intensity had statistical significance(P<0.05). The QDs-SA (1 045.4167±47.3623) was significantly higher than the mean fluorescence intensity of Cy3 (658.5467±55.0591). Conclusion QDs-SA fluorescence probes can effectively recognize APP and are sensitive and exceptionally photostable, suggesting that QDs-SA fluorescence probes could be a potential method in APP detection and offer a novel way for the diagnosis of Alzheimer's disease.

AIM: G protein-coupled inwardly rectifier potassium (GIRK) channel are distributed widely in mammalian brain. In CNS, GIRK 1/2 seems to be the predominant heterotetramers which play a pivot role in the regulation of the excitability of neurons and may contribute to the resting potential by leading to a hyperpolarization of membrane potential and reduction of the action potential frequency. In the context, the Weaver mouse is the first neurological abnormality directly linked to a genetic point mutation in the GIRK2 protein which includes spontaneous seizure. GIRK2 knock out mice showed normal development but more susceptible than normal mice to seizure induced by GABA antagonist. Here, we report that the mRNA and protein expression of GIRK subunit 2 is altered in kainic acid(KA)-induced epileptic rat hippocampus. METHODS: Rats were injected with kainate 14 mg/kg intraperitoneally to establish an acute and chronic temporal lobe epilepsy model. At chronic spontaneous seizure stage, by using of in situ hybridization, immunocytochemistry and Western blotting, the GIRK 1,2 mRNA and protein were analyzed quantitatively in the dentate gyreus, CA1, CA3 regions of hippocampus. RESULTS: GIRK1,2 mRNA and proteins were expressed abundantly in all regions of hippocampus. KA induced seizures and caused a significant increase in GIRK2 mRNA abundance and immunoreacitivity; only GIRK1 mRNA was increased significantly, but no difference was found by Western blotting protocol. CONCLUSION: GIRK1,2 mRNA and protein expression in the hippocampus of epileptic rat brain is up-regulated, which may be an adaptive response to over-excitability of neuron networks and prevent the over-excitability spread in hippocampus (DG-CA3-CA1). [

Objective To investigate the preventive effects of curcumin for status epilepticus(SE) induced by lithium chloride-pilocarpine.Methods Totally 45 Sprague-Dawley(SD) rats were randomly divided into three groups: preventive group(n=15),non-preventive group(n=15),and control group(n=15).The latency peroid and incidence of SE were recorded.The surviving neurons were stained by using nissl staining,and the programme death cells were detected by using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) in hippocampal CA3.Results The SE incidence of preventive group was 66.7%,which was significantly lower than that of non-preventive group(P0.05).Conclusion Pretreatment of curcumin can prevent the SE induced by lithium chloride-pilocarpine and the pretreatment can not protect the neuron.

Objective To investigate the effect of aerobic exercise combined with routine rehabilitation therapy on cardiorespiratory exercise function and rehabilitation efficacy in stroke patients with hemiplegia.Methods Thirty consecutive stroke patients with the first-ever hemiplegia were enrolled prospectively.They were randomly assigned to an experimental group (n=13) or a control group (n=13) according to the random number table (n=15 in each group).Four patients withdrew,and finally 26 patients completed the study (n=13 in each group).The two groups of patients were given routine rehabilitation therapy.On the basis of routine rehabilitation therapy,the experimental group was also treated with aerobic exercise training,once a day,30 min each time for 4 weeks.The evaluations of cardiopulmonary exercise tests were performed on both groups before treatment and after 4 weeks of treatment (peak oxygen uptake [VO2peak]),anaerobic threshold [AT],peak minute ventilation [VEpeak]),and motor function (Fugl-Meyer assessment scale [FMA],activities of daily living (Barthel index [BI]) were performed.The differences between the two groups before and after treatment were compared.Results (1)Cardiopulmonary exercise test results:Compared with before treatment,VO2peak,AT,and VEpeak were increased significantly (16±4 ml/[kg·min] vs.13±3 ml/[kg·min],12.9±2.5 kg·min vs.10.0±2.7 ml/[kg·min],41±12 L/min vs.34±9 L/min,t=9.668,4.328,and 3.662,respectively,all P<0.01).There were no significant differences in VO2peak,AT,and VEpeak (14±4 ml/[kg·min] vs.14±4 ml/[kg·min],11.8±2.6 ml/[kg·min] vs.11.5±2.9 ml/[kg·min],29±8 L/min vs.28±6 L/min) in patients of the control group between before treatment and after treatment.There were no significant differences (t=0.863,0.583,and 0.314,respectively,all P>0.05).The differences in VO2peak,AT,and VEpeak in the patients of the control group before and after treatment were 0.4 (0.2,0.6),0.1 (-0.5,0.7 ml/kg·min) and 0 (-2.5,5.0 L/min),respectively.The differences of VO2peak,AT,and VEpeak in patients of the experimental group before and after treatment were 3.0 (2.2,3.9 ml/[kg·min],2.7 (1.8,5.3 ml/[kg·min],and 7.0 (1.0,13.0 L/min),respectively.There were significant differences in VO2peak,AT,and VEpeak before and after treatment between the two groups (Z=4.287,3.721,and 2.236,respectively,P<0.05).(2) FMA score and BI evaluation results:Compared with before treatment,the FMA score and Barthel index after treatment in patients of the experiment group were 70±24 vs.52±28 and 78±20 vs.60±28 respectively,t=4.076 and 4.239 respectively,P<0.01).Compared with before treatment,the FMA score and BI in the control group after treatment were improved significantly (68±25 vs.60±27 and 69±22 vs.59±26 respectively,t=3.168 and 5.436 respectively,all P<0.05).The FMA score and the difference of BI in the control group before and after treatment were 5.0 (2.0,12.0) and 10.0 (5.0,15.0) respectively,and those of the experimental group before and after treatment were 10.0 (4.5,32.0) and 11.0 (10.0,27.5) respectively,and there were no significant differences between the two groups (Z=1.393,1.352,all P>0.05).Conclusion On the basis of routine rehabilitation therapy,stroke patients with hemiplegia increasing aerobic exercise training may improve their cardiopulmonary exercise function.

Aim To study the effect of melatonin on expression and activity of myosin light chain kinase in the artery wall of atherosclerotic rabbits.Methods The rabbit model of atherosclerosis was induced by a high-cholesterol diet.The blood lipid levels were assayed in the serum of each group.MLCK expression was detected by Western blot and immunohistochemical method.MLCK activity was measured by ?-32P-ATP incorporation into myosin light chain.Results The atherosclerosis model was established successfully.The levels of lipids decreased after MLT treatment.After fed with cholesterol for twelve weeks,the expression and activity of MLCK in the artery of atherosclerotic rabbits increased markedly,whereas there was no obvious difference in expression of MLCK in the artery of atherosclerotic rabbits fed with cholesterol and melatonin for twelve weeks compared with that of control.Conclusions It was suggest that high expression and activity of MLCK in the artery might be closely correlated with the development of atherosclerosis.Melatonin played an important role in inhibiting the development of atherosclerosis by decreasing the expression and activity of MLCK.