1.Interaction between mouse retinal astrocyte and antigen specific Th1 and Th17 Cells
Yan, CUI ; Hong-sheng, BI ; Sun DEMING
Chinese Journal of Experimental Ophthalmology 2012;30(1):25-28
Background C57BL/6andB10R Ⅲareroutinemurinespeciesusedinexperimental autoimmune uveitis (EAU).The inflammation is light for mouse after immunization whereas it is prominent for B10R Ⅲ.ObjectiveThis study was to observe the killing effect of interphotoreceptor retinoid binding protein (IRBP) 1-20-specific T cells on mouse retinal astrocyte.Th1 and Th17 cells effect in the EAU mechanism was discussed.MethodsB10RllⅢ mice and C57BL/6 mice were immunized with IRBP 161-180 and IRBP 1-20 in complete Freund adjuvant (CFA).The infiltrating cells of diseased B10R Ⅲ eyes were analyzed by flow cytometry.IRBP 1-20-specific T cells were isolated from the drainage lymph node and spleen and cultured in IL-2 or IL-23 for Th1 and Th17 cells polarization,respectively.Th1 and Th17 cells cultured for 5 days were seeded on the mouse retinal astrocyte monolayer pretreated with gamma interferon.Cell interaction was observed and the quantity of TNF-α was tested by ELISA.Every test was repeated 6 times and the mean was calculated.The maintenance of experimental animals complied with the Statement of ARVO.ResultsThere were lots of infiltrating cells in the eyes of B10Rm mice after immunization,including 9.5% IFNγ+ cells,5.1% IL-17+cells and 41.4% CD45+ cells.Six days after IRBP1-20 stimulation and cultured by IL-2 and IL-23,44.0% and 8.0% cells were IFNγ+,and 1.0% and 26.0% cells were IL17+.Twentyfour hours after the interaction between Th1 or Th17 and retinal astrocyte,retinal astrocyte died and detached.The killing effect of Th17 was stronger than Th1.48 hours after co-culture of Th1 or Th17T cells with astrocytes,the concentrations of TNF-α were ( 500± 10 ) and ( 801 ±24 μg/L) μg/L,respectively,with a significant statistical difference (t =-20.36,P =0.00).ConclusionsBoth Th1 and Th17 can kill retinal astrocyte,but Th17 plays a key role in the EAU pathogenesis process.The killing effect is caused by intercellular contact and interaction under the induction of cytokines.
4.Promoting proliferation and inhibiting apoptosis effects of sphingosine-1-phospate on human retinal pigment epithelium cells under the hypoxic condition
Yan, FAN ; Hong, LU ; Dingshan, HOU ; Wenjiao, BI ; Xiaomei, ZHANG
Chinese Journal of Experimental Ophthalmology 2015;33(1):33-37
Background Sphingosine-l-phospate (S1P) is a bioactive lipid and important messenger molecule in cells.It participates in the regulation of many biological processes,such as cell proliferation,migration,survival,differentiation,apoptosis,etc.Hypoxia is a trigger factor of choriod neovascularization (CNV) and pathological basis of many diseases,and retinal pigment epithelial (RPE) cells are involved in formation of CNV.However,the effects of S1P on proliferation and apoptosis of RPE cells are below understood.Objective This study was to investigate the influence of S1P on proliferation and apoptosis of human RPE cells under hypoxic conditions.Methods Human RPE cells line-D407 cells were cultured and passaged and generation 3-5 cells were used and divided into 6 groups.The cells were regularly cultured in the blank control group using DMEM containing 10% fetal bovine serum.CoCl2(200.00 μmol/L) was added into the colture medium for 2 hours in the hypooxic group.S1P of different concentrations (0.01,0.10,1.00,10.00 μmol/L) were added in culture medium 2 hours after the affection of 200.00 μmol/L CoCl2.The proliferative values of the cells were detected using WST-1 method as the absorbance (A value) and the proliferative rate of different groups were calculated.The apoptosis of the cells was assayed by Hoechst staining.The results were compared among different groups.Results Cultured cells showed the round-like in shape with clear nuclei and pigment.The proliferative values (A value) was 0.91 ±0.08,0.37±0.09,0.46±0.08,0.52±0.09,0.61 ±0.06,0.70±0.10 in the blank control group,hypoxic group and 0.01,0.10,1.00,10.00 μmol/L S1P groups,respectively,with a significant difference among the groups (F=21.104,P=0.000),and A values in various S1P groups were higher than those in the hypoxiac group (all at P<0.05).The proliferative rate was gradually raised with the increase of dose of S1P.Hoechst staining exhibited a few apoptosis cells in the blank control group,but in the hypoxic group,a lots of apoptosis cells were seen with the light-blue nuclei and condensable chromatin.However,the number of apoptosis cells was significantly decreased in various concentrations of S 1 P groups.The apoptosis rates were (1.21 ±0.08) %,(8.99 ±0.09) %,(6.60 ±0.08) %,(5.95 ±0.09) %,(4.81 ± 0.06)% and (3.96±0.10)% in the blank control group,hypoxic group and the 0.01,0.10,1.00,10.00 μmol/L S1P groups,respectively,with a significant difference among the groups (F =25.070,P =0.000).Compared with the hypoxia group,the cellular apoptosis rates of various S1P groups were lower (all at P<0.05).Conclusions Under the hypoxia condition,S1P can promote the proliferation of human RPE cells and inhibit apoptosis.
5.Autosomal dominant progressive external ophthalmoplegia,development of clinical symptoms in a Chinese family
Dao-Jun HONG ; Hong-Yan BI ; Ri-Liang ZHENG ; Xing-Hua LUAN ; Sheng YAO ; Yun YUAN ;
Chinese Journal of Neurology 2001;0(03):-
Objective To report the development of clinical symptoms in a Chinese family with autosomal dominant progressive external ophthalmoplegia(adPEO).Methods Electromyologram and muscle biopsy were performed in the proband and 4 family members with the disease.Results The proband was a 57 year-old woman,who developed bilateral ptosis after the age of 30,external ophthalmoplegia after the age of 35 years old,weakness of extremities at the age of 37 years old and bulb palsy with palmus at the age of 47 years old.In the family there were 20 male and female members from five generations.All of them complained about bilateral ptosis between 26—33 years old,external ophthalmoplegia(12/15)and weakness of all extremities(14/15)between 35—45,facial and masticatory weakness(9/9)as well as dysphagia(8/9)between 44—60,accompanied with heart lesions(4/7)after 50 years old.Some patients died due to cardiac impairment.Electromyologram showed myopathic abnormalities in the examined patients. The main myopathological changes were ragged red fibers,cytochrome c oxidase negative fibers and ragged blue fibers in succinate dehydrogenase staining.Conclusions The adPEO started from extra-ocular muscles to limbs,finally facial and bulbar muscles.Heart lesions were presented in late stage and lead to death in some members.The developing process of symptoms suggested that we should pay more attention to cardiac manifestations in this disease.
6.Role of plasma (1-->3)-beta-D-glucan in nephrotic syndrome complicated by fungous infection.
Xuan ZHANG ; Bi-Li ZHANG ; Wen-Hong WANG ; Yan LIU
Chinese Journal of Contemporary Pediatrics 2008;10(2):249-250
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Child, Preschool
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Female
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Humans
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Infant
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Male
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Mycoses
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blood
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diagnosis
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drug therapy
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Nephrotic Syndrome
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blood
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complications
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beta-Glucans
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blood
7.Self-assembly tissue engineering fibrocartilage model of goat temporomandibular joint disc.
Hong KANG ; Zhen-qiang LI ; Yan-da BI
West China Journal of Stomatology 2011;29(3):314-317
OBJECTIVETo construct self-assembly fibrocartilage model of goat temporomandibular joint disc and observe the biological characteristics of the self-assembled fibrocartilage constructs, further to provide a basis for tissue engineering of the temporomandibular joint disc and other fibrocartilage.
METHODSCells from temporomandibular joint discs of goats were harvested and cultured. 5.5 x 10(6) cells were seeded in each agarose well with diameter 5 mm x depth 10 mm, daily replace of medium, cultured for 2 weeks.
RESULTSOne day after seeding, goat temporomandibular joint disc cells in agarose wells were gathered and began to self-assemble into a disc-shaped base, then gradually turned into a round shape. When cultured for 2 weeks, hematoxylin-eosin staining was conducted and observed that cells were round and wrapped around by the matrix. Positive Safranin-O/fast green staining for glycosaminoglycans was observed throughout the entire constructs, and picro-sirius red staining was examined and distribution of numerous type I collagen was found. Immunohistochemistry staining demonstrated brown yellow particles in cytoplasm and around extracellular matrix, which showed self-assembly construct can produce type I collagen as native temporomandibular joint disc tissue.
CONCLUSIONProduction of extracellular matrix in self-assembly construct as native temporomandibular joint disc tissue indicates that the use of agarose wells to construct engineered temporomandibular joint disc will be possible and practicable.
Animals ; Cells, Cultured ; Collagen Type I ; Fibrocartilage ; Glycosaminoglycans ; Goats ; Temporomandibular Joint Disc ; Tissue Engineering
8.Synthesis and tumor resistance reverse activities ofα-hederagenin-type ring-A fused pyrazine derivatives
Xiao WANG ; Xian-Xuan LIU ; Yan-Ting YANG ; Hong-Bo WANG ; Yi BI
Chinese Journal of Pharmacology and Toxicology 2018;32(4):308-308
α-Hederagenin (H), derived from Hedera nepalensis var.sinensis, is a pentacyclic oleane-type triterpenoid that exhibits clear cytotoxicity to different tumor cell lines.In this study,a series of novel C-28 derivatives of hederagenin (H) were designed and synthesized in attempt to develop potent tumor resistance reverse activities agents. Previous research showed that H6 displayed robust reverse activity for paclitaxel resistance in KBV cells. Importantly, Co-treatment of paclitaxel with H6 significantly reduced the tumor weight to 42%. Pleasingly, H6 enhanced the efficacy of paclitaxel against KBV cancer cell-derived xenograft tumors in nude mice.Mechanism studies had found that H6 activated permeability glycoprotein(P-gp)ATPase,reduced intracellular ATP levels and inhibited efflux of P-gp substrates,thus enhancing the antitumor activity of paclitaxel on KBV cells.Molecular docking analysis of homology P-gp and H6 then conducted using the Surflex-Dock module.H6 showed a high binding affinity docking score with a total score of 5.4148,much higher than that of H(0.1414).The nov-el C-28 derivatives of H was synthesized from H6 via three-step reaction. The reversal activity of all synthesized H derivatives were tested using the MTT assay.The results showed that the derivatives of nitrogen groups at C-28 displayed same even potent activity than parent compound H6.In addition,its underlying mechanism of action and in vivo activity are in explore.
9.Pathological characteristics of skeletal muscle in patients with polymyalgia rheumatica
Hong-Yan BI ; Wei ZHANG ; Guang-Li SHEN ; He LV ; Sheng YAO ; Yun YUAN
Chinese Journal of Geriatrics 2003;0(11):-
Objective To study the pathological characteristics of skeletal muscle in patients with polymyalgia rheumatica(PMR).Methods Thirteen patients were diagnosed PMR according to the diagnostic criteria described by Bird in 1979.The onset of disease was between 49-78 years of age (median 60.3 years).All of them showed continuous symmetric myalgia with markedly elevated erythrocyte sedimentation rate(ESR).Five patients had mild fever and 8 patients had elevated C-reactive protein(CRP).Electromyogram(EMG)showed myogenic pattern in 2 patients.Only 1 patient showed anemia and 2 patients had mild elevated creatinine phosphokinase.None of them showed evidence of temporal arteritis.Biceps brachii muscle biopsies were performed in all patients before corticosteroid therapy.Results The typeⅡmuscle fiber atrophy was observed in all patients.Moth-eaten changes appeared in 8 cases,increased intramuscular lipid drops in 8 cases and ragged red fibers(RRF)in 3 cases of them.After steroid therapy,all patients showed improvement of myalgia and normal ESR as well as CRP.Conclusions The main symptoms of the patients are myalgia and abnormal ESR.Some patients have high level of CRP and abnormal EMG.Anemia and temporal arteritis are less frequent.The main myopathological changes were typeⅡmuscle fiber atrophy.Some patients have moth-eaten changes.More lipid drops and RRF indicate abnormal metabolism of skeletal muscle.
10.Clinical features of mtDNA A3243G mutation in adult patients
Sheng YAO ; Ri-Liang ZHENG ; Hong-Yan BI ; Zhao-Xia WANG ; Yun YUAN ;
Chinese Journal of Neurology 2000;0(04):-
Objective To analysis the clinical manifestations of mtDNA A3243G mutation in adulthood.Methods The clinical features were investigated in 36 cases (28 patients from 5 families with the mutation and 8 sporadic cases),in whom mtDNA A3243G mutation was confirmed genetically in 23 cases (15 cases from 5 mutation families and 8 sporadic cases).Cranium radiology was performed in 14 cases.Muscal biopsies were performed in l0 cases.Results Among 28 cases in the 5 family,there were 9 cases (32.1%) with stroke like episodes,17 cases (60.7%) with diabetic mellitus and 16 cases (57.1%) with deafness.Such symptoms usually combined with each other and rarely existed alone. Cardiomyopathy and renal failure were uncommon.In the 23 cases with mtDNA A3243G mutation,14 cases (61.0%) had mitochondria] myopathy,encephalopathy,lactic acidosis,and stroke-like episodes (MELAS),mostly presenting cognitive abnormalities,dysarthria or aphasia and headache,3 cases (13.0%) were asymptomatic carriers,2 cases (8.7%) had autonomic dysfunction,2 cases (8.7%) had diabetic mellitus with or without nerve deafness,1 case (4.3%) had diabetic mellitus with infertilitas and cardiomyopathy,respectively.Cranial radiological images revealed the changes more commonly in the temporal and occipital lobes and less frequently in the frontal lobes.Ragged red fibers were confirmed in 9 of 10 cases with muscle biopsies.The proportion of mutant mtDNA A3243C was not significantly different between MEALS (28.75%?13.69%) and non-MELAS (25.08%?11.54%).Conclusions mtDNA A3243G mutation mainly results in the lesions in the central nerve system,pancreatic island and acoustic nerve in adulthood.Heart and kidney are less frequently involved.Cognitive abnormalities,aphasia and headache are the major symptoms of adult MELAS.Families have with more than 1 patient with diabetic mellitus and deafness,indicating that the mutation is other than MELAS mutation.We should pay more attention to the non-MELAS symptoms in the families with mtDNA A3243G mutation.