1.Trans-scleral ocular drug delivery for the treatment of posterior segment eye diseases
Chinese Journal of Ocular Fundus Diseases 2017;33(2):213-217
The human sclera accounts for 95% of the surface of the eyeball,providing ample contact area which is suitable for targeted trans-scleral ocular drug delivery.Currently there are several tans-scleral sustained-release strategies,including intra-scleral delivery,episcleral delivery,as well as tans-scleral iontophoresis.Different devices and methods have their own advantages and disadvantages,for example,intrascleral delivery is somehow invasive,and episcleral delivery device needs to be made thin to prevent erosion of conjunctiva,iontophoresis needs to be frequently repeated as of its short-term effect.With the development of bio-material engineering technology,episcleral microfilm could become an ideal drug delivery route for posterior segment ocular diseases.
2.Suprachoroidal space drug delivery C
Chinese Journal of Ocular Fundus Diseases 2017;33(2):209-213
The suprachoroidal space (SCS) is the potential space between the sclera and choroid.Drugs delivered through SCS can bypass the sclera,avoiding clearance by conjunctival and scleral blood vessels and lymphatic circulation,so that more drugs can reach the disease tissues such as choroid and retina.SCS drug delivery does not disrupt the ocular integrity,is safer than the intravitreal drug injection and more effective than trans-scleral drug delivery.In addition,SCS delivery only needs a very small volume of drug,which makes it possible to be carried out in multiple parts of the sclera,and the specific disease area can be more precisely targeted.SCS drug delivery is suitable for the treatment of choroidal and retinal diseases.However,currently SCS drug delivery is still a novel field and many aspects need to be more in-depth studied,including its safety,delivery methods,drug formulation and effectiveness.
3.Diagnosis and management of complications in nephrotic syndrome
Liping ZHAO ; Lingyun YANG ; Yun CHENG
Chinese Journal of Applied Clinical Pediatrics 2016;31(5):333-336
Nephrotic syndrome(NS)is the most frequent cause of glomerular disease in children,it can deve-lop many complications,including infection,thrombosis and acute kidney injury,while acute adrenocortical suppression is a complication of some drugs administration. This review focuses on the pathophysiology and management of common complications in patients with NS.
4.Heterochromia in rabbit-heterochromia in a dutch pigmented rabbit
Mitsuko TOYOGUCHI ; Lingyun CHENG ; William R FREEMAN
International Eye Science 2007;7(4):932-933
· Heterochromia was observed in a six-month-old Dutch pigmented rabbit and the rabbit was examined for general and eye anomalies. The rabbit showed a blue eye with fundus hypochromia on the right and a brown eye with partial fundus hypochromia on the left. White fur (white forelock) was present, but deafness was not apparent although no objective audiologic examination was performed. Histology studies of both eyes revealed that significantly fewer pigment cells in iris stroma and less pigmentation in retinal pigment epithelium and choroid in the right eye than in the left eye.
5.The use and future of transgenic animals in pharmacological research
Lingyun LI ; Ting WANG ; Guoxiang CHENG ;
Chinese Pharmacological Bulletin 1987;0(03):-
Transgenic animals are those in which recombinant DNA technology is used to remove a gene or a specific part of DNA with exogenous DNA, which is hereditable and can be expressed stably. Transgenic rodent technology, first demonstrated by Gordon in 1980,opened the door to the next step in biotechnology and gene regulation, which has been used to reproduce or mimic a part of a human disease in a complex cellular environment by creating in vivo models with germ line transmission of the introduced genetic regulatory elements. Animal models for drug discovery are developing increasingly, which is promoting the research and development new drugs greatly. The use and development of transgenic animals in pharmacological research is reviewed in this article.
6.Cytotoxicity of dimethyl sulphoxide on ocular cells in vitro
Chunhui, ZHAO ; Bifei, LAN ; Jiangping, HOU ; Lingyun, CHENG
Chinese Journal of Experimental Ophthalmology 2015;33(3):216-220
Background Dimethyl sulfoxide(DMSO) is a commonly used adjuvant to promote testing drug solubility to prepare multi-levels testing drug concentrations.DMSO is cell type-dependent cytotoxic and its toxicity can interfere the testing drug evaluation.Determining its safe concentration on commonly used cell types is important for ocular drug development.Objective This study was to determine the minimal toxic concentration of DMSO for in vitro ocular cell lines in a simulated drug screening setting.Methods Retinal pigment epithelial (RPE) cells were isolated from one pigmented rabbit and primarily cultured.Human RPE cell strain (ARPE19),scleral fibroblasts line (S75-Fron),human Müller cell line (MIO-M1),human lens epithelial cell line (HLEC),human choroidal melanoma cell line (OCM-1),human umbilical endothelial cell (HUVEC) and human HeLa cell line (HELA) were cultured.Different concentrations of DMSO (1.6%,1.0%,0.8%,0.4%,0.2% and 0.1%) were prepared with 160 μl DMSO solution and 9.84 ml RPMI1640 (or DMEM/F12 or DMEM) containing 2% fetal bovine serum.Different concentrations of DMSO were added in medium for 96 hours,and the and viability (absorbance) of the cells was detected using MTS to evaluate the cytotoxicity of DMSO.Results Rabbit primary RPE cells showed the yellow-green fluorescence for cytokeratin(CK) and HMB45 red fluorescence for S100.The viability of the cells was gradually declined as the increase of DMSO dose,showing significant differences in ARPE19,S75-Fron,HLEC,OCM-1,HUVEC and primary RPE cells (all at P<0.05),and when DMSO concentrations were ≥ 0.8%,the cell viabilities were significantly lower.But no significant difference was found in MIO-M1 cells among different doses of DMSO (F=0.830,P=0.547).The minimal toxic concentration of DMSO for ARPE19,HUVEC,HELA,HLEC,MIO-M1,OCM-1,primary RPE cells and S75-Fron was 0.8%,0.1%,0.8%,>1.6%,>1.6%,0.2%,0.2%,0.2%,respectively,and HUVEC was more sensitive to the cytotoxicity of DMSO (P=0.02),and MIO-M1 was the least sensitive to DMSO (P =0.39).The viability of HUVEC and primary RPE cells went down with the increase of DMSO dose,and S75-Fron viability started to decline in 0.1% DMSO and then stabilize with the higher concentrations until 1.6% DMSO at which the viability showed further decline.Conclusions The tolerability of ocular cells in vitro to DMSO varies depending on the cell types.The minimal toxic concentration ranged from 0.1% to 1.6%.The result suggests that a concurrent DMSO control should be set up along with the testing compound.
7.Effect of human hypoxia inducible factor-1? gene transfection on apoptosis of A?25-35 induced hippocampal neurons of primary culture
Lingyun LIU ; Xiqing CHAI ; Qingjun LI ; Guihua CHENG
Journal of Third Military Medical University 2003;0(07):-
Objective To construct the eukaryotic expression vector for human hypoxia inducible factor-1? gene (pSNAV-HIF-1?), and to investigate the apoptosis and intracellular calcium concentration of the transfected A?25-35 induced hippocampal neurons of primary culture. Methods Human hypoxia inducible factor-1? gene from pBSKhHIF1?T7 was inserted into pSNAV2.0 by the method of gene recombination, then the constructed vector was transfected into hippocampal neurons of primary culture and followed by A?25-35 for treatment. Empty pSNAV2.0 vector was treated the same way as pSNAV-HIF-1? as a control. The expression level of HIF-1? protein was assayed by Western blotting. Apoptosis was detected by flow cytometry. Intracellular calcium concentration was determined by laser scanning confocal microscopy with Fluo-3/AM as the fluorescent dye. Results It was shown that pSNAV-HIF-1? was successfully constructed by restriction enzyme digestion, PCR and DNA sequencing. The expression level of HIF-1? protein was significantly increased in transfected hippocampal neurons of primary culture (P
8.Echocardiographic diagnosis of parachute mitral valve
Cheng YU ; Mingxing XIE ; Lingyun FANG ; Ke LI ; Yali YANG ; Lin HE
Chinese Journal of Ultrasonography 2014;23(1):16-19
Objective To explore the value of echocardiography in the diagnosis of the parachute mitral valve(PMV).Methods The echocardiographic characteristics of 12 patients with PMV between 2005 and 2012 were reviewed retrospectively,including the morphology,movement and blood flow of mitral valve apparatus,and other associated cardiac abnormalities.Results Of the mitral valve leaflets in these 12 cases,all were thickened and opened restricted in diastole.Axial view showed that in all cases mitral valves opened eccentrically and in 2 cases they didn't close tightly.Each patient only had single papillary muscle in left ventricle,which received all chords of mitral valves.Of papillary muscles in these patients,4 were located at posteromedial wall,3 at anterolateral wall,2 at middle posterior wall,2 at apical wall,and 1 at posterolateral wall.Of the 12 patients,only one were simple PMV,11 had associated cardiovascular anomalies,2 had mild regurgitation in systolic period and 11 had increased trans-valve peak flow velocity.Conclusions Echocardiography is reliable in the diagnosis of PMV,which could provide a comprehensive evaluation of valvular lesions and other associated cardiovascular anomalies.
9.Effects of umbilical cord-derived mesenchymal stem cells on matrix metalloproteinases secreted by fibroblast-like synoviocytes from rheumatoid arthritis
Lu ZHANG ; Wei KONG ; Yile REN ; Xuebing FENG ; Cheng ZHAO ; Lingyun SUN
Chinese Journal of Rheumatology 2014;(10):665-669
Objective To explore the effects of umbilical cord-derived mesenchymal stem cells (UCMSCs) on fibroblast-like synoviocytes(FLSs) from patients with rheumatoid arthritis(RA). Methods collagen-induced arthritis(CIA) models were developed on Wistar rats and 1 ×106 UCMSCs were given by intravenous injection from tail vein on the 17th day. On day 42, rats were sacrificed and synovial tissues were obtained to detect matrix metalloproteinase (MMP)-1, MMP-3 and MMP-13. Synovial tissues from patients with RA and osteoarthritis(OA) treated by knee arthroplasty were used to isolate FLSs. RNA of FLSs were extracted to compare MMP-1, MMP-3 and MMP-13 expression. FLSs and UCMSCs were cocultured through transwell for 72 hours. Then levels of MMPs were compared in the supernatants by Luminex. The MMPs expressed by FLSs and soluble factors expressed by MSCs were detected by real time-polymerase chain reaction(PCR). After adding antibodies to soluble factors, the MMPs expressions of RA FLSs were compared. Results MMP-1 (6.9±5.4, 1.3±1.4, P<0.05), MMP-3 (6.0±6.5, 1.4±1.0, P<0.05) and MMP-13 (21.8± 20.8, 1.5±1.6, P<0.05) expression were much higher in CIA rats compared with healthy controls. MMP-1 (1.3±1.4, 6.9±5.4,8.7±6.8, P<0.05), MMP-3(1.4±1.5, 6.0±6.5, 6.0±5.7, P<0.05) and MMP-13(3.0±3.2,
22±21, 22±26, P<0.05) expression were inhibited by UCMSCs in vivo. In vitro, MMP-1 (1.8±0.9, 0.9±0.7, t=2.44, P<0.05), MMP-3(2.6±1.7, 1.1±1.0, t=2.25, P<0.05) and MMP-13(2.4±2.3, 0.6±0.7, t=2.37, P<0.05) levels were higher in RA than OA FLSs. After coculture, MMP-13(1.3±1.2, 0.9±1.2, t=3.63, P<0.05) expressed by FLSs were down-regulated, however MMP-1 (1.5±1.4, 6.6±6.0, t=3.90, P<0.05) and MMP-3 (7±17, 22±35, t=2.86, P<0.05) were up-regulated when analyzed by paired t-test. Soluble factors such as I-DO, HGF and IL-10 were elevated. Anti-IL-10 antibody could decrease the function of UCMSCs by inhibiting MMP-13 expression in RA FLSs. Conclusion UCMSCs ameliorates RA by secreting soluble factor IL-10, which may inhibit MMPs expressed by FLSs.
10.Establishment of a visualized detection method of Sendai virus by reverse transcription loop-mediated isothermal amplification
Jie ZHOU ; Lijuan ZHAO ; Lingyun TAO ; Liju NI ; Cheng GAO ; Hongyan CHEN
Acta Laboratorium Animalis Scientia Sinica 2016;24(3):293-298
Objective To establish a simple and sensitive detection method of Sendai virus ( SeV ) by reverse transcription loop-mediated isothermal amplification ( RT-LAMP) technique. Methods According to the published Gen-Bank sequences (DQ219803. 1), six pairs of primers were designed targeting the conserved region of SeV. The amplifica-tion products were detected with a LAMP real-time Turbidimeter. (LA-302). Through optimizing the LAMP primers and re-action conditions, a rapid and specific detection method of SeV was established. Meanwhile, the amplified products were colored by fluorescence detection reagent after completion of the reaction, so that the amplification could be visualized and detected by naked eyes. Then, methodological evaluation of the RT-LAMP was tested. Results The method of RT-LAMP showed a highly efficient amplification for SeV viral target gene which was performed at 63℃ for 60 min with the LAMP re-al-time Turbidimeter (LA-302). The detection limit was 2. 1 TCID50, 100 times higher than that of RT-PCR, and no cross-reaction with other RNA and DNA viruses of mice was observed. The results of SeV LAMP reaction was visualized and the tube could be directly observed by naked eyes with the addition of fluorescence detection reagent. The results were consist-ent with the results detected by real-time tubidimeter. 92 clinical samples were detected byRT- LAMP, RT-PCR and indi- rect ELISA, and the coincidence rate was 100%. Conclusions This established SeV RT-LAMP detection method is fast, specific, highly sensitive,easy to perform under simple conditions, and is suitable for rapid detection of Sendai viirus.