Background To establish the ideal animal model of retinitis pigmentosa (RP) is very important for onward relevant study.Previous research determined that N-methyl-N-nitrosourea (MNU) can selectively damage photoreceptors via intravenous injection in mammal.However,whether MNU can be used to create an RP model needs to be investigated.Objective This experiment was designed to evaluate the toxic effect of MNU on photoreceptor cells of cats.Methods MNU was injected into 20 2-year-old cats via femoral vein and randomized into 20mg/kg, 25mg/kg, 30mg/kg, 35mg/kg and 40mg/kg MNU groups,and equal amount of normal saline solution was used in the same way in 4 normal cats as the control group.The activity,pupil size and light reflex were observed after injection of MNU.The cats were sacrificed and eyeballs were enucleated for histological examination to evaluate the structural and morphological changes of photoreceptors at 24 hours,72 hours,7 days and 14 days after the administration of MNU.This experimental study complied with the Statement for the Use of Animals in Ophthalmic and Vision Research.Results Dilated pupil and inertia of light reaction were found in experimental cats on the 7th days in the various groups.In 24 hours after MNU injection,the damage of photoreceptors was primarily characterized by pyknosis and disorder.In 72 hours after MNU injection,attenuation of the outer nuclear layer and disruption of cells were seen.Loss of photoreceptors and disappearance of the outer nuclear layer were observed on the 7th and 14th day.The extent of retinal photoreceptor cell damage was dependent on the dose of MNU.Conclusion MNU can selectively induce serious damage of the photoreceptor cells in cats retina in a time- and dose-dependent manner.

Background Many studies and clinical trials of pharmacologic vitreolysis are already under way to try to improve vitreo-retinal surgery and to liquefy and detach the vitreous from the retina ultimately, including chondroitinase,hyaluronidase,dispase and plasmin. However, there has not been any report on purification of human plasminogen from cord blood plasma and inducing posterior vitreous detachment of the animal eye at present.Objective This study was designed to isolate and purify the production of human plasminogen (Plg) from cord blood plasma with ethanol precipitation and evaluate the efficacy of Plg in inducing posterior vitreous detachment (PVD).Methods Human Plg was Separated and purified from cord blood plasma by ethanol precipitation method. The protein band corresponding to Plg with molecular mass of 92 000 was revealed in SDS-PAGE and confirmed by MALDI-TOF and Mascot database. Anion-exchange chromatography and plasminogen activity assay kit were used to obtain purified Plg with biological activity. Twenty-five fresh pig eyes were enucleated and assigned to 5 groups and 5 eyes for each group. The normal eyes were used as control group. Balanced salt solution(BSS)of 0.1 ml was intravitreally group and standard substance group. All of the eyes were then incubatedfor 60 minutes under the 37 ℃. Retinal histopathology and ultrastructure were examined under the light microscopy, scanning electron microscopy ( SEM ) and transmission electron microscopy (TEM). Results The Plg with potential fibrinolytic activity was successfully extracted and purified from cord blood plasma by ethanol precipitation method. No posterior vitreous detachment (PVD) was seen in normal control group, BSS group and r-SK group following the intravitreal injection under the sem. However,PVD was demonstrated in r-SK+ Plg group and standard substance group under the SEM. The inner limiting membrane ( ILM ) and the retina were well preserved in all of the experimental eyes. No retinal morphology and ultrastructural abnormality were found under the light and SEM and TEM. Conclusion Ethanol precipitation is a feasible way to isolate and purify Plg from human cord blood plasma. Extracted Plg shows potential fibrinolytic intravitreal injection of Plg.

Accidents, Occupational ; Adolescent ; Adult ; Ammonia ; poisoning ; Female ; Humans ; Male ; Young Adult

Objective To establish a method for rapidly detecting the G-protein ?3 subunit (GNB3) 825 site single nucleotide polymorphism (SNP) and to analyse the relationship between GNB3 825 site gene SNP and obesity. Methods The real-time fluorescent PCR was employed to analyse the GNB3 825 site gene SNP of 420 samples from 21 provinces and the the frequencies of genotypes were compared with those detected by gene sequencing. GNB3 825 site genotype, body weight, body mass index (BMI) and fat content were examined from 207 subjects and the correlation between GNB3 825 site gene SNP and obesity was analysed. Results The result by real-time fluorescent PCR showed that the frequencies of 825T and 825C haploid were 46.90% and 53.10%, respectively, and the frequencies of 825TT, 825TC and 825CC genotype were 22.38%, 51.42% and 28.10%, respectively, with no other genotype detected, which was consistent with the result by gene sequencing. BMI and fat content were significantly higher in subjects with GNB3 825TT than in subjects with other genotypes. Body weight was much higher in subjects with GNB3 825TT genotype than in subjects with 825CC genotype, but not significantly different with 825CT genotype. Conclusion A new rapid method for the detection of GNB3 825 site SNP has been successfully established. There existed significant correlation between GNB3 825TT genotype and obesity.

Diagnosis, Differential ; Female ; Humans ; Knee Joint ; pathology ; Middle Aged ; Sarcoma, Synovial ; pathology ; Tomography, X-Ray Computed

To develop gastric floating erodible plug pulse capsules with compound Danshen as the model drug, in order to realize the pulse release of traditional Chinese medicines. Through the study on impermeable capsules, optimized prescriptions, drug-containing rapid-release tablets and prescription screening, and erodible plug prescription and process, we successfully prepared compounded Danshen pulse capsule, so as to provide a new dosage form for controlling and treating heart disease to better cater to clinical demands.
Capsules ; Delayed-Action Preparations ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Heart Diseases ; drug therapy ; Permeability ; Salvia miltiorrhiza ; chemistry

Child ; Contracture ; congenital ; therapy ; Humans ; Male ; Neck Muscles ; pathology ; Torticollis ; congenital

Background Bevacizumab is primarily aimed at pathologic angiogenesis for off-label uses such as the treatment of ocular neovascular disorders.However,as a new anti-fibrotic and anti-angiogenic agent following trabeculectomy,the safety and efficacy of bevacizumab by multiple-time and high-dose subconjunctival injection are still under study.Objective This study was to assess the safety and efficacy of bevacizumab after multiple-time and high-dose subconjunctival injections.Methods Regular trabeculectomy filtration surgery was performed on both eyes of 18 clean New Zealand White rabbits 0.1ml of bevacizumab(25g/L) was subconjunctivally injected intraoperatively and 3,5,7 days postoperatively in the left eyes of rabbits,and no any intervene in the right eyes were as normal controls.Bleb morphology was examined every 2 days and graded based on Moorefield's criteria and compared between the bevacizumab-treated eyes and normal saline(NS) eyes.The animals were sacrificed at 10,20 and 30 days after surgery respectively.The histopathological changes of the blebs were detected by hematoxylin and eosin stain to evaluate the cellular element around the bleb,and Masson stain was used to assess the degree of fibroblast proliferation.The degree of vascularity of bleb was identified by anti-vWf stain.Approval of this protocol was obtained and permitted from People's Hospital Institutional Animal Care and Use Committee of Peking University.The use of experimental animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Compared to the NS-treated eyes,bevacizumab-treated eyes showed the larger and more diffusely elevated blebs with the significant difference(2.48±0.22cm2 versus 1.73±0.27cm2,t=5.194,P＜0.05).The survival time of the filtration bleb in bevacizumab panel was longer in bevacizumab-treated eyes compared to control eyes,showing a significantly difference between them(21.0±1.56 days versus 12.5±1.97 days,t=3.830,P=0.005).Histological and immunohistochemical analysis confirmed that bleb and adjacent conjunctiva vascularity(A value) was significantly less in bevacizumab-treated eyes than that in control eyes at 20 days after surgery with the difference value 14320.7±4134.9(t=12.275,P＜0.05),and fibroblast deposition value was evidently diminished after bevacizumab treatment at 30 days following surgery in comparison with control eyes with the mean difference 0.27±0.03(t=15.980,P＜0.05＝.Conclusion Repeated subconjunctival injection of bevacizumab can effectively prolong the survival time of bleb in a rabbit model of trabeculectomy and limit the degree and area of vascularization in 30 days following surgery.Bevacizumab inhibit fibroblast-meditated tissue formation significantly in the later phase of vascularization after trabeculectomy.

Background Macular hole in high myopia has been paid tremendous attention in clinical research due to its high recurrence rate and unpredictable prognosis.Spectral domain optical coherence tomography (SD-OCT) is considered to be of a good evaluating value for macular hole,yet its application is compromised in patient with high myopia.High myopia might lead to retinal detachment which consequently prevents an accurate OCT.Therefore,it is important to assess the effectiveness of OCT on prognosis in these patients after surgical restoration of the detached retina.Objective The aim of this study was to evaluate the risk of reopening of a macular hole in highly myopic patient after removing silicone oil by SD-OCT.Methods A case-observational study was designed.Twenty-five highly myopic patients with monocular macular holes who underwent vitrectomy combined with internal limiting membrane peeling and silicone oil tamponade were included in this study.Four patients were male and 21 patients were female,with the average age (61.4±9.0) years old and diopter (-14.14 ±6.86)D.Regular ocular examination,axial length measurement,fundus photography and OCT were performed at the day before removing silicone oil and every month till the 6 months after operation.Retinal thickness,macular height index (MHI) and choroidal thickness were measured by OCT.Written informed consent was obtained from each patient.Results All 25 patients finished the follow-up of 6 months.Macular holes remained closure 6 months after operation in 19 patients (76％) and reopened in 6 patients macular holes were in 1-5 months (24％),with a mean time at (3.3± 1.4) months.The average foveal retinal thickness,MHI and the choroidal thickness at the middle of the macular hole were (216.5±95.6) μm,0.30 ± 0.09 and (122.9 ± 20.5) μm in the closed group,respectively,and those in the unclosed group were (113.5±28.7) μm,0.58±0.27 and (96.8±22.9) μm,with significant differences between the two groups (t=2.577,-4.143,2.669,P ＜ 0.05).The percentage of macular hole closure was 85.7％ in the MHI＜0.5 group and 25.0％ in the MHI ≥ 0.5 group,showing a significant difference (P =0.031).The base diameter in the unclosed hole group was significantly larger than that in the closed hole group (1070.2±393.6 μm versus 533.3±277.7 μm) (t =-3.700,P =0.001).Conclusions The measurements of the retinal thickness,MHI and the choroidal thickness at the central area of the macular hole are helpful for the evaluation of reopening risk of macular hole after removing silicone oil.

Objective To explore the role of CD 30 on CD4 + T lymphocyte and T lymphocyte subset activation in asthma pathogenesis.Methods Twenty seven asthma active attack patients,16 acute upper respiratory infection patients and 19 control children were randomly selected. Direct immunofluorescence flow cytometry was used to detect the CD 30 positive percentage on CD4 +cell; ELISA was used to detect the levels of interleukin(IL) 4 and IL 13 in culture supernatants of peripheral blood mononuclear cells(PBMC) and plasma total immunoglobulin E(IgE) level.Results Compared with control group and acute upper respiratory infection group, in asthmatic children the CD4 +CD 30 + percentage on CD4 + cell was elevated significantly; 2.The levels of IL 4 and IL 13 in supernatants of cultured lymphocyte were increased significantly, the plasma total IgE level was increased significantly;3.There was a significant positive correlation of CD4 +CD 30 + cell percentage with the levels of IL 4 and IL 13 in culture supernatants and plasma total IgE.Conclusions The cell that could produce Th2 derived cytokine may consist of CD 30 +cell;When CD 30 L combined with CD 30 on CD4 + cells ,it might result in Th2 cell proliferation ,develop and release Th2 derived cytokine; IL 4 and IL 13 could induce B cell production and secrete more IgE. It is suggested that CD 30 and imbalance of Th2 subset may play an important role in asthma pathogenesis.