1.Cellular growth under hydrostatic pressure using bovine aortic EC-SMC co-cultured ePTFE vascular graft.
Lei SUN ; Koichi NIWA ; Jian-zhong LIN ; Takeshi KARINO
Journal of Zhejiang University. Science. B 2005;6(2):79-82
High blood pressure (hypertension) is implicated in the development of atherosclerosis. Blood vessels are constantly subjected to stretch due to blood pressure and changes in stretch usually instigate adaptive vascular remodeling, including abnormal growth and proliferation of vascular smooth muscle cells (VSMCs) as well as extracellular matrix (ECM). In this experiment, we used bovine aortic endothelial cells and smooth muscle cells (EC-SMC) co-cultured ePTFE vascular grafts subjected to normal atmospheric pressure (as a control), and 100 mmHg hydrostatic pressure for 7 d. The increase of cell layer thickness was observed. When measured, the cell layer thickness increased by 116.2%. The increase of collagen (Type IV) synthesis was also observed in the immunohistochemistry assay. When stained with toluidine blue, the cells showed metachromatic phenomenon.
Animals
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Blood Vessel Prosthesis
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Cattle
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Cell Culture Techniques
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methods
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Cell Proliferation
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Cells, Cultured
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Collagen Type IV
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metabolism
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Hydrostatic Pressure
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Mechanotransduction, Cellular
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physiology
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Muscle, Smooth, Vascular
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cytology
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physiology
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Polytetrafluoroethylene
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chemistry
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Tissue Engineering
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methods
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Transplants
2.Early signs of cognitive impairment in patients with obstructive sleep apnea hypopnea syndrome: an event-related potential study.
Ke ZOU ; Yuanfeng SUN ; Xiangdong TANG ; Fei LEI ; Lina DU ; Zhesi CHEN ; Tingting YAN ; Zhong ZHENG
Journal of Biomedical Engineering 2014;31(4):870-874
This study seeks to explore the early signs of cognitive impairment in patients with obstructive sleep apnea hypopnea syndrome (OSAHS). According to polysomnography, twenty patients diagnosed with OSAHS and twenty normal controls underwent event-related potential (ERP) examination including mismatch negativity (MMN) and P300. Compared with normal controls, OSAHS patients showed significantly prolonged latency of MMN and P300 at Cz. After controlling age and body mass index (BMI), MMN latency positively correlated with apnea hypopnea index (AHI), oxygen reduction index, stage N1 sleep and arousal index, while MMN latency negatively correlated with stage N3 sleep and mean blood oxygen saturation; and P300 latency positively related to AHI and oxygen reduction index; no relationships were found among MMN latency, MMN amplitude, P300 latency and P300 amplitude. These results suggest that the brain function of automatic processing and controlled processing aere impaired in OSAHS patients, and these dysfunction are correlated with nocturnal repeatedly hypoxemia and sleep structure disturbance.
Case-Control Studies
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Cognition Disorders
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complications
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physiopathology
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Event-Related Potentials, P300
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Humans
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Hypoxia
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physiopathology
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Oximetry
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Polysomnography
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Sleep Apnea, Obstructive
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complications
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physiopathology
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Sleep Stages
3.HPLC fingerprint of glycyrrhizea radix et rhizoma praeparata cum melle.
Lei SUN ; Yong JIN ; Xiao-Qing LIU ; Shan-Yi QIAO ; Song GAO ; Yan-Zhong CHE
China Journal of Chinese Materia Medica 2014;39(11):2056-2059
The chromatographic fingerprint was established by eluting with the mobile phase consisted of acetonitrile and 0.2% formic acid water on an Agilent TC-C18 (2) column (4.6 mm x 250 mm, 5 microm). Six chromatographic peaks were identified by HPLC-MS/MS method. Ten batches of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle were determined, and the similarity was arranged from 0.72 to 0.99. Good precision, stability and repeatability were obtained, and this study provides a reference for the quality control of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle.
China
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Chromatography, High Pressure Liquid
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methods
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Drugs, Chinese Herbal
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chemistry
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Glycyrrhiza uralensis
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chemistry
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Mass Spectrometry
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Quality Control
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Rhizome
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chemistry
4.Pizhen therapy for the treatment of neck pain caused by cervical spondylosis: a report of 61 cases.
China Journal of Orthopaedics and Traumatology 2008;21(6):459-460
Acupuncture Therapy
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methods
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Adult
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Aged
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Cervical Vertebrae
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Female
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Humans
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Male
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Middle Aged
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Neck Pain
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therapy
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Spondylosis
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therapy
5.Investigation on the correlation between incidence rate of high uric acid and biochemical indicators of organ lesions among hospitalized children
Ling WU ; Yazhen DI ; Yuanling CHEN ; Shiling ZHONG ; Lei HE ; Nan SUN
Chinese Journal of Rheumatology 2014;18(9):632-636
Objective To explore the correlation between the distribution of uric acid (UA) level and the biochemical indicators which reflect the degree of organ lesions among hospitalized children.Methods Patients who were hospitalized to the Department of Pediatrics and received the blood UA test from June 2012 to October 2013 were included,23 217 cases in total.The biochemical analyzer-Japan's Olympus AU 2700 was used to detect blood biochemistry; and uricase-peroxidase coupling method was used to detect blood UA.Among these patients,2 099 cases whose UA level exceeded the normal level.Then the patients' gender,age,primary diagnosis and UA level were recorded; and the distribution of their UA level was described.The Chi-square or Fisher test was used to assess the incidence rate.At the same time,each blood biochemical indicators of the patients with high UA level were recorded ; and the relationships between the blood UA of the 1 650 patients with complete records and each blood biochemical indicator were analyzed with Pearson correlation analysis.Results The incidence rate of hyperuricemia among the hospitalized children was 9.04%(2 099/23 217).In particular,the incidence rate among boys and girls was 6.5t%(890/13 657) and 12.65% (1 209/9 560) respectively (x2 =256.9,P<0.05).The incidence rate of hyperuricemiin different diseases was as follows:in the critical illness 36.93% (113/306),neonatal disease 20.34% (922/4 533),urinary system diseases 12.08% (47/389),circulatory system diseases 11.67% (21/180),nervous system diseases 11.05%(112/1 014),digestive diseases 10.50%(190/1 810),infectious diseases 10.18%(120/1 179),blood diseases 7.58% (55/726),endocrine system diseases 5.74% (17/296),autoimmune diseases 4.24% (48/1 131),respiratory diseases 3.90% (454/11 653) respectively (x2=1423.0,P<0.05).The incidence of hyperuricemia at younger than one month was 18.31%(929/5 075),younger than one year old was 4.22% (359/8 501),younger than six years old was 10.68%(600/5 618),younger than 15 years old was 5.24% (211/4 023) respectively (x2=858.5,P<0.05).Blood UA was positively correlated to urea nitrogen,creatinine,lacticdehydrogenase,α-hydroxy-butyrate dehydrogenase,creatine kinase and creatine kinase-MB (r=0.426,0.44,0.324,0.367,0.413,0.431,P<0.05).Blood UA was not correlated to fructosamine,blood glucose,triglycerides,total cholesterol,low-density lipoprotein and high density lipoprotein.Conclusion The incidence of hyper-uricemia among hospitalized children is high; and the incidence among children with severe diseases and newborn babies is high; followed by in children with urinary system and circulatory system diseases.The blood UA level is closely related to the blood biochemical indicators which reflect the lesions of heart and kidney.
6.Research on diagnosis and surgical treatment of congenital coronary artery fistula
Chunsheng LI ; Zhongya YAN ; Zhong LU ; Zhengyan ZHU ; Hong LEI ; Yun SUN ; Guangcun CHENG
Chinese Journal of Postgraduates of Medicine 2016;39(3):217-220
Objective To summarize and analyze clinical diagnosis and surgical treatment methods of 11 cases with congenital coronary arterial fistula (CCAF). Methods The clinical data of 11 patients who were definited by ultrasonic cardiogram, CT angiography (CTA) and coronary angiography were analyzed retrospectively. Four cases were simple CCAF, 7 cases coexisted with other heart abnormalities. Six cases were given surgical closure of fistula without cardiopulmonary bypass. One of the cases adopted coronary artery under the tangent cotton-padded mattress suture, and 4 cases fistula arterial were ligatured directly. Six cases were given surgical closure of fistula under cardiopulmonary bypass. The right coronary arterial was opened in three of the cases with right coronary artery aneurysm to close fistula. The chambers of heart in the others were opened to close fistula. Results All patients received surgical treatment successfully, and no death happened during the operation. The ultrasonic cardiography showed that all patients recovered well. Follow-up was conducted on 10 patients with the time period ranging from 3 months to 5 years. There was no death and no complication. Conclusions Combined application of ultrasonic cardiogram, coronary angiography and CTA increases the accuracy rate of diagnosis greatly and offers visual bases to formulate operation plan. Surgical operation is quite effective for congenital coronary arterial fistula after definite diagnosis. Operator should try to reserve the expanded coronary arterial, strengthen the anticoagulant after opeation to prevent thrombosis.
7.Experimental study on the eyelid reconstruction in situ with the silk fibroin membrane
Lei, ZHONG ; Xiao-feng, ZHANG ; Zheng-tai, SUN ; Jing, XIA ; Ying-ming, WANG
Chinese Journal of Experimental Ophthalmology 2011;29(11):968-972
Background Autologous and allograft renal transplantation exist some disadvantages of less donor source and rejection.As a scaffold of cell in tissue engineering,fibroin was determined to have a good biocompatibility.But whether the fibroin membrane can become a substitution for tissue defect is seldom reported.Objective This experiment aimed to investigate the feasibility of silk fibroin membrane in the rabbit eyelid reconstruction in situ.Methods A 4 mmx3 mm tarsi defect model was created on the upper eyelids of 18 healthy New Zealand white rabbits.The eyelid reconstruction in situ was performed with regenerated silk fibroin membrane material in the right upper eyelids (silk fibroin group ) and allogenic sclera material (sclera group ) on the upper eyelids of fellow eyes.The grafts were clinically examined for the evaluation of inflammation and implant exposure at the first,second and forth week after operation.The inflammation response and collagen distribution were examined by hematoxylin & eosin staining and Masson staining.Expression of basic fibroblast growth factor(bFGF) in the grafts was detected by immunohistochemistry,and ImagePro Plus software was used for statistical analysis.Results All eyelid defects showed a primary healing.The surface of palpebral conjunctival was smooth and the inflammation of ocular surface was mild.The eyelid margin in the sclera group was more notch than that in the silk fibroin group.Results of pathological examination revealed that the arrangement of collagen fibers in the sclera group was more disordered,but that in the silk fibroin group was regular.The expression level(A value) of b-FGF in the operative area in silk fibroin group were 0.027 67±0.004 69,0.051 73±0.008 72,0.058 72±0.006 88,and those in the sclera group were 0.056 48±0.009 14,0.072 83 ± 0.009 17 and 0.078 73 ±0.010 84 in 1,2,4 weeks after operation,showing statistically significant differences between two groups in various time points ( t =- 6.38,t =- 4.99,t =- 2.87,P <0.05 ).Conclusions Silk fibroin membrane can reconstruct the eyelid shape in situ with the less inflammation response and good biocompatibility.Silk fibroin membrane could be used to support the eyelid as a new tarsal repairing materials.
8.Influence of monochromatic light exposure on the growth of Müller cell and the expression of myopia-related cell factors in vitro
Jing, XIA ; Xiao-feng, ZHANG ; Wei, XIA ; Lei, ZHONG ; Zheng-tai, SUN ; Ying-ming, WANG
Chinese Journal of Experimental Ophthalmology 2013;(5):446-451
Background Research showed that exposure of 530 nm monochromatic light can induce myopia in animal,and retinal Müller cells participate in the formation of myopia.However,the effect and mechanism of retinal Müller cells during the formation of monochromatic light induced-myopia is below understood.Objective This study was to investigate biologic characteristics of rat retina Müller cells and the expression of cell factors in Müller cells after being illuminated by the 530 nm monochromatic light,and discuss the role of the retina Müller cells in myopia induced by monochromatic light.Methods Immortalized rat retinal Müller cells were cultured with DMEM containing 10% fetal bovine serum in a self-made cell incubator with monochromatic light by adjusting luminance of 530 nm LED source.The cells were exposed to 125,250 and 500 lx luminance respectively for 6,12 and 24 hours,and the cells without light-irradiation were used as control.The growth of the cells under the different light time and different illuminations was described by MTT as the absorbance at the wavelength 570 nm (A570),and cell cycle analysis of Müller cells was performed by flow cytometry 48 hours after cultured,and the expression of transforming growth factor-beta 1 (TGF-β1),tyrosine hydroxylase(TH),inducible nitric oxide synthase(iNOS) and basic fibroblast growth factor(bFGF)in the cells were detected by reverse transcription PCR(RT-PCR),respectively.Results The Müller cells were uniform in size with polygonal shape and defined edges.No statistically significant difference was found in the A570 value in the cells of the 125 lx and 250 lx illuminated groups compared with the control group in various time points(P>0.05).However,significant lowing was seen in the A570 value in the cells of the 500 lx illuminating for 12 hours and 24 hours in comparison with the control group (P =0.013,0.001).Compared with the control group,the ratio of the number between G2 and G1 phase was not significantly declined in 125 lx,250 lx illuminating for 48 hours (P =0.073,0.330),and the ratio in the 500 lx illuminating group was significantly lower than those in the 250 lx illuminated group and the control group (P =0.028,0.038).RT-PCR revealed that the expression of TGF-β1 mRNA in the cells was higher in the 250 lx illuminated group than that of the 500 lx illuminated group (P=0.006).The expression of iNOS mRNA was gradually upregulated in the 250 lx illuminated group compared with the control group (P =0.001),but that in the 500 lx illuminated group was downregulated (P =0.000).The expression of bFGF mRNA was raised in the 125 lx and 250 lx groups but reduced in the 500 lx group when compared with the control group(P=0.002,0.000,0.005).Also,the expression of TH mRNA was significantly increased in the 250 lx group(P=0.000),but decreased in the 500 lx group(P=0.000,P=0.001).Conclusions The monochromatic light of 530 nm can inhibit the growth of rat Müller cells and downregulate the expression of myopia-related cell factors and therefore exert effect in the formation of myopia.
9.Expression of inflammatory factors in conjunctiva and assessment of dry-eye associated examination in ovariectomized rat
Lei, ZHONG ; Xiaofeng, ZHANG ; Jing, XIA ; Zhengtai, SUN ; Yingming, WANG ; Wei, XIA
Chinese Journal of Experimental Ophthalmology 2014;32(9):807-812
Background Dry eye is increasing gradually recently,but its etiology and manifestation are very diverse.Studies showed that menopause of adult females was one of the risk factors of dry eye.In addition,some inflammatory factors also participate in the pathogenesis and development.But the study on the relationship of sex hormone with inflammation and ocular surface damage is still below understanding.Objective This study was to investigate the expressing changes of interleukin (IL) and tumor necrosis factor-α (TNF-α) in conjunctiva and the manifestation of ocular surface in ovariectomized rat model.Methods Twenty clear female SD rats were randomized into the ovariectomized group and the sham operative group according to randomized number table.Ovariectomy was performed in the ovariectomized group,and abdominal myotomy without ovariectomy was performed in the sham operative group.Serum estrogen and androgen levels were detected by radiation immunoassay 3 months after operation.Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescence staining were carried out in the rats before operation and 1 month,2 and 3 months after operation.The morphology of conjunctival epithelial cells was examined by hematoxylin & eosin staining at the 3rd month after operation.The expressions of IL-17A,IL-1 β,IL-6 and TNF-α in conjunctiva were semi-quantitative analyzed by immunohistochemistry and Western blot.The use and care of the animals complied with State Science and Technology Commission Regulations for the Administration of Affair Concerning Experimental Animals.Results Serum estrogen levels were (23.53 ± 1.65) pg/L and (47.89 ± 1.05) pg/L 3 months after surgery in the ovariectomized group and the sham operative group,respectively; the serum androgen levels were (1.84±0.09) ng/L and (2.47±0.12)ng/L in the ovariectomized group and the sham operative group,respectively,showing a significant decline of serum estrogen and androgen levels in the ovariectomized group compared with the sham operative group (t=-35.37,-12.13,both at P<0.01).No significant differences were seen in S Ⅰ t between the two groups among various time points (Fgroup =0.38,P =0.55 ; Ftime =0.13,P =0.72 ; Finteraction =0.39,P =0.76).No obvious fluorescence staining was seen in the cornea of both the ovariectomized group and the sham operative group.The histopathological examination showed that the layers of rat conjunctival epithelial cells increased with the disordered arrangement in the ovariectomized group.Immunochemistry showed that the expressions of IL-17A,IL-1β,IL-6 and TNF-α (A values) were significantly higher in the ovariectomized group than those in the sham operative group (IL-17A:t=8.22,P<0.01 ;IL-1β:t=16.43,P<0.01 ;IL-6:t=13.44,P<0.01 ;TNF-α:t=16.26,P<0.01).Western blot assay showed the similar results (IL-17A:t=19.41,P<0.01 ;IL-1β:t=12.63,P<0.01 ;IL-6:t=17.17,P<0.01 ;TNF-α:t=15.19,P<0.01).Conclusions Serum estrogen and androgen levels drop obviously,and there is an up-regulation of IL and TNF-α expression in conjunctiva tissue in the ovariectromized SD rats.However,no obvious dry eye-related sign occurs.
10.Inhibition of FK506 on the expression of vascular endothelial growth factor in retinal Müller cells cultured by high concentration glucose
Wei, XIA ; Jing, XIA ; Xiaofeng, ZHANG ; Lei, ZHONG ; Zhengtai, SUN ; Yingming, WANG
Chinese Journal of Experimental Ophthalmology 2013;32(11):998-1003
Background Retinal Muller cells participate in the pathological process of diabetic retinopathy (DR) through expressing vascular endothelial growth factor (VEGF).It is reported that FK506 inhibits the expression of VEGF in solid tumors and experimental corneal neovascularization,but whether FK506 exerts its role on retinal Müller cells or not is still unclear.Objective This study aimed to investigate how FK506 affects the expression vascular endothelial growth factor (VEGF) in rat retinal Müller cells under the condition of high glucose.Methods Immortalized rat retinal Müller cell line was regularly cultivated and logarithmic phase of cells were incubated in 96-well plate with the cell density of 1 × 104/ml.Different concentrations of FK506 (800.00,400.00,200.00,100.00,75.00,50.00,25.00,12.50 and 6.25 pg/ml) (100 μl/well) were added into the culture medium to determine the half maximal inhibitory concentration (IC50) of FK506 by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT).The cell lines were cultured with DMEM medium (containing D-glucose of 5.5 mmol/L) or high glucose DMEM (containing D-glucose of 50 mmol/L),and 75 pg/ml FK506 were added into DMEM,respectively,and the cells were divided into the normal control group,FK506 group,high glucose culture group and high glucose + FK506 group.ELISA was employed to assay the content of VEGF protein in the cell supernatant.The expressions of VEGF mRNA and protein in the cells were detected by reverse transcription PCR (RT-PCR) and Western blot,respectively.Results The cells grew well in the normal control group,FK506 group,high glucose culture group and high glucose+FK506 group in 12,24 and 48 hours after culture with the polygon-like shape.The IC50 of FK506 was 75 pg/ml.The contents of FK506 in the cell supernatant were (966.46± 13.59) pg/ml,(1 059.42±67.43) pg/ml,(16 243.11 ±3 926.38) pg/ml and (9 467.25± 1 525.56) pg/ml in the normal control group,FK506 group,high glucose culture group and high glucose+FK506 group,respectively,showing a significant difference among the four groups (F =20.51,P =0.00).The VEGF levels in cell supernatant were significantly higher in the high glucose group than those of the normal group and the high FK506 group (P =0.00,P =0.02),but no significant difference was found in the VEGF level in cell supernatant between the control group and FK506 group (P =0.08).The expressions of VEGF mRNA and protein in the cells were significantly different among the four groups (F=126.06,P=0.00;F=5.44,P=0.01),and the relative expressing values of VEGF mRNA and protein in the cells of the high glucose group were significantly higher than those of the control group and the high+ FK506 group (all at P<0.01).The relative expressing values of VEGF mRNA and protein were 0.64±0.09 and 0.68±0.18 in the FK506 group,which were lower than those of the normal control group (0.84±0.07 and 0.75± 0.03).However,no significant differences were seen between the two groups (P =0.05,0.07).Conclusions The expression of VEGF in rat retinal Müller cells up-regulates under the high glucose condition.FK506 plays inhibitory effects on VEGF expression to certain extent in vitro.