Objective To induce the experimental corneal neovascularization(CRNV) by alkali burn,and explore the methods for quantitative analysis of CRNV and observation of permeability of CRNV. Methods Thirty-six C57BL/6 mice were selected and divided into experiment group(n=30) and control group(n=6).For the experiment group,alkali burn was induced by application of filter paper with 1mol/L sodium hydroxide to the cornea for 5 s.For the control group,no intervention was conducted.Areas of CRNV were measured on day 4,7,10 and 14 after alkali burn.Histological examinations of cornea were performed with HE staining on day 3, 7,10,16 and 28 after alkali burn.On day 10,endothelial cell marker CD31 was used with immunohistochemical staining for CRNV counting,and fluorescence angiography(FA) was employed to reveal the permeability of CRNV.Corneal ulceration and hyphema were observed everyday.Results CRNV developed after alkali burn,and extincted afterwards.Axenic coneal ulceration and hyphema were frequently observed,with the incidences of 6.7% and 10.0%,respectively.Histologic changes of corneal tissues at different time points could be observed with HE staining.On day 10, CRNV could be labeled and counted with immunohistochemical staining for CD31 antibody,and the permeability of CRNV could be detected by FA. ConclusionCRNV counting with immunohistochemical staining for CD31 antibody and measurement of area of CRNV are appropriate methods for quantitative analysis of CRNV.FA is an effective method in the detection of permeability of CRNV.

Objective To evaluate the role of mitochondrial permeability transition pore (mPTP) in attenuation of myocardial ischemia-reperfusion (I/R) injury by sufentanil postconditioning in rats.Methods Sixty male Sprague-Dawley rats,aged 14-15 weeks,weighing 350-420 g,were randomly divided into 4 groups (n =15 each):sham operation group (group S),group I/R,cyclosporin A group (group CP) and sufentanil postconditioning group (group SP).Myocardial I/R was produced by occlusion of left anterior descending branch of coronary artcry for 30 min followed by reperfusion.In groups CP and SP,cyclosporin A 5 mg/kg and sufentanil 1 μg/kg were injected via the jugular vein at 5 min before reperfusion respectively,while the equal volume of normal saline was injected in group I/R.At 10 min of reperfusion,hearts were excised,the myocardial mitochondria were immediately isolated and the activity of mPTP was measured by spectrophotometry.MAP and HR were recorded at 30 min of equilibration,at 30 min of ischemia and at 120 min of reperfusion and rate-pressure product (RPP) was calculated.Arterial blood samples were obtained at 120 min of reperfusion for determination of the plasma cardiac troponin Ⅰ (cTnⅠ) concentration.The animals were then sacrificed for determination of infarct size (IS) and area at risk (AAR),and IS/AAR was calculated.The mitochondrial ultra-structure was examined with electron microscope.Results Compared with group S,the mPTP activity and plasma cTnI concentration were significantly increased,and MAP and RPP were significantly decreased in the other three groups (P ＜ 0.05).Compared with group I/R,the mPTP activity,plasma cTnI concentration and IS/ARR were significantly decreased in groups CP and SP,and MAP was significantly increased in group CP (P ＜ 0.05).Compared with group CP,MAP was significantly decreased (P ＜ 0.05),while no significant change was found in the other indexes in group SP (P ＞0.05).Significant mitochondrial swelling,and disruption and disappearance of cristae were showed in I/R group.The mitochondrial structure was more complete in CP and SP groups than that in group I/R,and the disrupted cristae were found in a small number of mitochondria in CP and SP groups.Conclusion The mechanism by which sufentanil postconditioning reduces myocardial I/R injury is related to inhibition of mPTP opening in rats.

Objective To investigate the effect of type 2 diabetes mellitus (DM) on the attenuation of myocardial ischemia-reperfusion (I/R) injury by sufentanil postconditioning in rats.Methods Male pathogen-free Sprague-Dawley rats,weighing 160-180 g,were used in the study.A model for type 2 DM was established by the feeding of high-fat diet-induced insulin resistance and intraperitoneal streptozocin 35 mg/kg.DM was confirmed by blood glucose level ≥ 16.7 mmol/L one week after injection.Eighteen type 2 diabetic rats were randomly divided into 3 groups (n =6 each):DM sham operation group (DM-S group) ; DM-I/R group; DM sufentanil postconditioning group (DM-SP group).Another 18 healthy nondiabetic rats were chosen and randomly divided into 3 groups (n =6 each):nondiabetes mellitus sham operation group (NDM-S group) ; nondiabetes mellitus I/R group (NDM-I/R group) ; nondiabetes mellitus sufentanil postconditioning group (NDM-SP group).Myocardial I/R was induced by 30 min occlusion of the left anterior descending branch of coronary artery (after 30 min of equilibration) followed by 120 min of reperfusion.Sufentanil 1.0 μg/kg was injected via the right jugular vein 5 min before reperfusion in NDM-SP and DM-SP groups.MAP,SP and HR were recorded immediately before ischemia,at 30 min of ischemia and at 120 min of reperfusion and rate-pressure product (RPP) was calculated.Arterial blood samples were collected at 120 min of reperfusion for measurement of plasma cardiac troponin Ⅰ (cTnⅠ) concentration.The rats were then sacrificed for determination of the myocardial infract size.Results MAP and RPP were decreased,while the plasma cTnl concentration was increased during reperfusion in diabetic and nondiabetic rats.Sufentanil postconditioning decreased the myocardial infract size and plasma concentrations of cTnⅠ,and increased MAP and RPP during reperfusion in nondiabetic rats,but had no effect on the parameters in diabetic rats.Conclusion Type 2 DM interferes with sufentanil postconditioning-induced myocardial protection in rats.

Objective To investigate the association between endothelial dysfunction and arterial stiffness in continuous ambulatory peritoneal dialysis (CAPD) patients.Methods Ninetyfour stable CAPD patients from a single center were enrolled in this cross-sectional study.Ultrasound evaluation was conducted on brachial artery to estimate endothelial-dependent flow-mediated dilation (FMD).Automatice pulse wave velocity (PWV) measuring system was applied to examine the carotidfemoral PWV.Blood pressure and biochemical parameters were detected.Pearson's correlation and Stepwise multiple regression analysis were performed to explore the relationship between FMD and PWV.Results PWV was significantly higher in patients with diabetes as compared to those without diabetes[(13.25± 1.66) m/s vs (11.24±1.92) m/s,P ＜ 0.01].Furthermore,PWV was positively correlated with age(r=0.319,P=0.002),SBP (r=0.289,P=0.005) and C-reactive protein (r=0.211,P=0.041),was negatively correlated with albumin (r =-0.429,P =0.001) and FMD (r=-0.466,P=0.001).In multivariate regression analysis,diabetes mellitus,albumin,FMD,age and SBP were independently associated with PWV after adjustment.Conclusion Endothelial dysfunction is associated with greater arterial stiffness in CAPD patients.

Objective To explore the clinical characteristics,therapy reactions and prognosis of the elderly patients with idiopathic thrombocytopenic purpura(ITP). Methods A total of 43elderly ITP patients(age≥60 years old)including 16 men and 27 women were reviewed and further followed up for 1 month to 15 years. Results Until June 2007,35 elderly ITP patients survived,platelet counts were sustained(30-50)×109/L in 7 cases,but no significant bleeding was found.Thirty-six patients had adrenocorticosteroid therapy first, 25 patients were sensitive to adrenocorticosteroid therapy,4 patients underwent splenectomy,and 3 patients achieved a normal platelet count. Immunosuppressive agents(vinscristine,cyclophosphamide, azathioprine and Cyclosporin A)treatments were held in 5 6 case-times,Cyclosporin A and azathioprine were more effective than vinscristine and cyclophosphamide.The refractory rate was 13.9%.One patient progressed to monoclonal gammopathy of unknown significance and 1 to lymphoma.Eight patients died.1 patient died of brain bleeding after trauma,3 patients died of malignant neoplasm,4 patients died of heart failure induced by infection. Conclusions The clinical features of elderly ITP patients are atypical.the mortal bleeding in them was rare,treatment strategy should be individualized tO each elderly patient.

Objective To investigate the brain metabolic changes and evaluate their spatial distributions after traumatic axonal injury (TAI)in rats by using proton magnetic resonance spectroscopy(1H-MRS).Methods The TAI model was made by subjecting the head of the rats to the linear and angular accelerations.The multi-voxel MRS was employed to detect the tissue metabolic state at the levels of hippocampus-caudate and pons prior to injury and at 24 hours after injury.The alterations of NAA/Cr,NAA/Cho and Cho/Cr values as well as the spatial distribution of NAA/Cr reduction were accessed. Immunohistochemical staining for β-APP was used to observe the injured axons. Results A siguificantdecrease in NAA/Cr and NAA/Cho(P<0.05)and subtle increase in Cho/Cr(P>0.05)were observed in rats at 24 hours after TAI in comparison to the pre-injury levels.Notable decrease in NAA/Cr value was observed in the areas including the brain stem,hippocampus,internal capsule,corpus callosum and thalamus,where axonal injuries were confirmed by the histological examination. Conclusion Metabolic imbalances Occur in the brains of rats with TAI.with notable changes in the brain stem and the hippocampus.

Objective To study the effectiveness and feasibility of low intensity of aerobic bicycle ergometer training on hemiplegic stroke patients unable to perform assistive or independent ambulation.Methods Thirty stroke patients unable of assistive or independent ambulation were randomly divided into a control group and a aerobic training group.Patients in the aerobic training group performed low intensity of aerobic bicycle ergometer training for 30 min,3 times a week for 6 weeks in addition to a routine rehabilitation regime.The training duration and course of routine rehabilitation training were the same in both groups.The related cardiovascular response in aerobic training and changes in motor performance as well as the risk factors for stroke were observed in both groups.Results Twenty-four patients(12 patients in each group)completed the experiment.Patients in aerobic training group performed all training regime safely without any adverse response.Their motor function(including FMA score,Rivermead index,BI ADL ability),exercise endurance duration in exercise test,glucose level at 2 h in oral glucose tolerance test,fasting insulin level and HOMA-IR index all improved greatly as compared to the control group(P ＜ 0.05).Conclusion Low intensity of aerobic bicycle ergometer training can be safely and effectively administered in hemiplegic stroke patients unable of assistive ambulation or independent ambulation.

Objective:To explore the role of hippocampal γ oscillation abnormality in sepsis-associated encephalopathy (SAE).Methods:Seventy male Sprague-Dawley rats (2-3 months) were randomly (random number) divided into three groups according to the random digital table method: sham, CLP, and CLP + dopamine 4 (D4) receptor agonists RO-10-5824 group. The SAE animal model was established by cecal ligation and puncture (CLP). On day 10-14 after surgery, the open field, novel object recognition, and fear conditioning tests were performed. After that, the hippocampus was collected to measure expressions of parvalbumin (PV) and D4 receptor. In another set of experiment, CA1 local field potential (LFP) were recorded, and the relationship between LFP and time with novel object was analyzed. Independent sample t-test was used for pairwise comparisons, and multiple comparisons were performed by one-way ANOVA, followed by the Tukey multiple comparisons test. Correlation was analyzed using Pearson correlation. Statistical significance was assumed when P<0.05. Results:Compared with the sham group, hippocampal PV (77.54±4.61)%, D4 expression (56.36±3.88)% and γ oscillation power (41.1±8.62)%, object exposure time (36±3) s, new object recognition rate (49±4)%, and scene stiffness time (56±7) s were decreased significantly ( P<0.05). However, RO-10-5824 treatment could increase hippocaml γ oscillation power (92.3±6.7)%, and reverse the decreased new object exposure time (44±3) s and new object recognition rate (63±4)%. Correlation analysis showed that hippocampal γ oscillation power was positively associated with new object exposure time ( r=0.609 2, P=0.015 9). There was no difference in total distance traveled or time spent in the center among groups ( P>0.05). Conclusion:Hippocampal γ oscillation abnormality might play a key role in cognitive impairment associated with SAE.

Objective To investigate the expression and clinical significance of CXCR3/CXCL10in human cancer.Methods CXCR3 and CXCL10 were detected in 60 paraffinic tissues of patients with primary breast cancer,20 of mammary fibroma and 20 of mastopathy by immunohistochemistry S-P method and two stage method.Results The expression of CXCR3 (40/60,66.7％ ) and CXCL10 (45/60,75％)in breast cancer was higher than that in mastopathy [CXCR3(8/20,40％ )x2 =4.44,P =0.035 ;CXCL10( 10/20,50％ )x2 =4.36,P =0.037)].The expression of CXCR3 was related to status of axillary lymph node metastasis,clinical stage and the expression of HER-2 (x2 =4.15,P =0.042; x2 =7.74,P =0.021 ;x2 =4.27,P =0.039).The expression of CXCR3 had positive relationship to the number axillary lymph node metastasis( rs =0.375,P =0.003 ),clinical stage ( rs =0.451,P =0.000).Conclusions CXCR3 may be related to the progression and metastasis of breast cancer,and it may be used as a marker of breast cancer prognosis.

OBJECTIVETo report the results of clinical characteristics, enzyme activity determination and mutation analysis of GLB1 gene in a Chinese patient with mucopolysaccharidosis (MPS) type IVB (Morquio B disease).

METHODA 14-year-old Chinese boy with MPS type IVB was firstly diagnosed by blood leucocytes galactosamine-6-sulfate sulfatase (GALNS) and β-galactosidase (GLB1) determination, who was characterized by short stature, multiplex skeletal abnormalities, difficulty in walking. PCR-sequencing analysis was applied to detect the mutations in GLB1 of the patient.

RESULTThe patient was characterized by dwarfism, pectus carinatum, kyphosis, normal intelligence, and no neurologic damage of spasms, linguistic capacity and so on. The patient had normal GALNS enzyme activity and very low GLB1 enzyme activity [5.03 nmol/(h·mg) vs. normal value 118 - 413 nmol/(h·mg) ] in leukocytes. A compound heterozygous missense mutations c.442C > T(p.R148C)/c.1454A > G(p.Y485C) in GLB1 gene were detected in this patient. The mutation p.Y485C is a novel variant. With the method of gene analysis of new variant, the mutation p.Y485C was considered to be a pathogenic mutation.

CONCLUSIONThe MPS IVB patient showed severe multiple skeletal deformities, normal intelligence, no neurologic damage and very low GLB1 enzyme activity, who carries compound heterozygous mutations p.R148C/p.Y485C. The mutation p.Y485C in GLB1 gene may be a novel pathologic mutation of MPS type IVB.

Adolescent ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Chondroitinsulfatases ; genetics ; metabolism ; DNA Mutational Analysis ; Humans ; Joints ; pathology ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis IV ; enzymology ; genetics ; pathology ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Radiography ; Spine ; diagnostic imaging ; pathology ; beta-Galactosidase ; genetics ; metabolism