The therapeutic response of anti-vascular endothelial growth factor (VEGF) differs among individuals.According to the changes of central retinal thickness,intraretinal fluid,subretinal fluid,best corrected visual acuity and other morphological or functional manifestations after treatment,the performance of the treated eyes can be classified as optimal response,poor response and non-response.A variety of factors could account for poor or non-response to anti-VEGF,such as genomic polymorphism and specific genomic risk alleles,lesion characteristics,vitreous and macular structural abnormalities,resistance to anti-VEGF drug,and the role of pericytes and others.The common counter measures include increasing the dosage,shortening the injection interval and replacing with another alternative drug,inhibition of pericytes,relieving vitreomacular anatomical abnormalities.It is still worthy of further exploration that how to assess individual reasons for non-response,so that we can give proper treatment to reduce the excessive use of anti-VEGF drugs and improve the clinical management of ocular neovascularization diseases.

Objective To investigate the risk factors of renal damage in children with Henoch-Schonlein purpura(HSP).Methods Two hundred and ninety-two children with HSP admitted to the First Affiliated Hospital of Anhui Medical University from Jan.2007 to Mar.2008 were reviewed.According to the findings urinary examination,children were divided into non-renal damage group(183 cases)and renal damage group(109 cases).Clinical data(gender,age,predisposing factors,rash repeated frequency,system damage)and laboratory examination [blood WBC,PLT,IgA,fibrinogen(FIB),TT,ASO,C3,C4] of 2 groups were compared,and then multiple regression analysis were performed to evaluate the risk factors of renal damage in children with HSP.Results 1.Among them,37.33% cases had renal da-mage,and hematuria and proteinuria manifestations were common.2.Simple Logistic regression showed that age was at least 8 years old and the number of purpura relapses was at least 4 times,increased serum FIB levels had significant difference between the 2 groups(P=0.030,0.000 and 0.011,respectively).There was no significant difference between the 2 groups in other clinical data and laboratory examinations.3.Mutiple Logistic regression showed that when the number of purpura relapses was at least 4 times,increased serum FIB levels were the risk factors of the renal damage(P=0.000 and 0.029,respectively);while there was no significant difference for the children who were at least 8 years old between the 2 groups(P=0.175).Conclusion When the number of purpura relapses is at least 4 times,increased serum FIB levels are the risk factors of the renal damage in children with HSP.

Objective To explore the possible association of oxidative stress reaction with the mRNA expression of podocin in rats with adriarnycin(ADR)-induced nephrotic syndrome.Methods mRNA expression of podocin in renal cortex were investigated by in situ hybridization staining and semi-quantitative RT-PCR in ADR-induced nephrotic rats,the level of malondialdehyde(MDA), superoxide dismutase(SOD)and total antioxidative capacity(T-AOC) in renal cortex were measured, then the relationship between them was evaluated.Results In ADR rats, MDA increased at d7 and reached a significant higher level at d14 and d21;SOD decreased at d14 and persisted to d21; T-AOC decreased at d21.The podocin mRNA mostly expressed in cytoplasm of glomerular cells. Both the number and the intensity of positive cells increased notably as time progressed in ADR rats.The level of podocin mRNA expression showed no obvious changes at d7, while prominently increased at d14 and pesisted to d21 in ADR rats.There was significant correlativity between the mRNA expression of podocin and the level of MDA,SOD,T-AOC.Conclusion The mRNA expression of podocin is correlated with oxidative stress reaction in rats with ADR-induced nephrotic syndrome.

Objective To investigate the alteration of protein kinase C (PKC) and endothelin system in early diabetic rats, and the effect of specific PKC inhibitor on the expression of retinal endothelin-1 (ET-1). Methods The rats model with streptozotocin(STZ)-induced diabetes were set up. The expression of retinal PKC was detected by enzyme-linked immunoabsorbent assay (ELISA). The expression of retinal ET-1, ET-3, ET-A and ET-B receptor mRNA was determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The alteration of retinal ET-1 mRNA after intravitreal injection of PKC inhibitor GF109203X in diabetic rats was also observed. Results The activities of membranous PKC were significantly increased in 2-week diabetic rats compared with that in normal rats(t=3.296, P=0 008), while activities of cytosolic PKC were unchangeable(t=0 138, P=0 894). The expression of retinal ET-1 mRNA was significantly increased(P=0 008), while no change was found in expression of ET-3, ET-A and ET-B mRNA(P=0 918,P=0 889,P=0 500). After intravitreal injection of 10 -5、10 -6、10 -7 mol/L PKC inhibitor GF109203X in diabetic rats, the expression of retinal ET-1 mRNA was decreased in a dose-dependent manner compared with the control rats. Conclusion Activation of PKC and increased expression of ET-1 could be found in the retina of early diabetic rats, and PKC inhibitor could inhibit the expression of retinal ET-1.

Objective To evaluate the effect of single hand pipe method on venous indwelling needle puncture. Methods Literature about the impact of single hand pipe method on venous indwelling needle puncture was retrieved from digital databases of PubMed, Medline, EMbase, The Cochrane Library, SinoMed, VIP database, China National Knowledge Infrastructure(CNKI), and WanFang Data. The quality of literature was evaluated by the Cochrane Handbook (5.1.0). The Meta data was analyzed by RevMan 5.3. Results Ten random control trials were included . Ten studies indicated that single hand pipe method increased success rate of puncture[related to the risk(RR)=1.16, 95%confidence interval(CI) 1.11-1.22]. And, it decreased the complication rate (3 studies) (RR=0.32, 95%CI 0.13-0.79) and pain rate(2 studies) (RR=0.25, 95%CI 0.13-0.47) as well as shorten the time of tube(2 studies) (WMD:-1.68, 95%CI-2.44--0.93). Conclusions Single hand pipe method can improve a puncture success rate and reduce complications and pain rate and shorten the time of tube, so that get more time to rescue the patient, and improve the level of the working efficiency and nursing for the nursing staff. In addition, to some extent, save manpower. It is worth being popularized.

Objective To probe a selective cultural method for bovine retinal endothelial cells (BREC) and pericytes (BRP) in vitro. Methods With the isolation of active retinal blood vessels, BREC were cultured in a fibronectin coated substrate and Dulbecco′s Modified Eagle′s Medium (DMEM) supplemented with 10% human serum and 100 ?g/ml heparin, while homogeneous cultures of retinal pericytes were obtained when isolated microvessels were seeded to uncoated dishes and grown in DMEM supplemented with 20% fetal bovine serum. BREC were identified by acetylated-low density lipoprotein (Dil-Ac-LDL) incorporation and immunohistochemical method of Von Willebrand factor, while BRP were identified by immunohistochemical method of ?-isoform of smooth-muscle actin. Results The purity of selectively cultured BREC and BRP was more than 98%, being reproducible. BREC got together around the microvessel fragments with the small-cyprinoid-like configuration at first,and could phagocytize Dil-Ac-LDL with the expression of fluorescence in cytoplasm. The expressions of Von Wllebrand factor and ?-isoform of smooth-muscle actin were positive and negative respectively in BREC, while were negative and positive respectively in BRP. Conclusion BREC and BRP with high purity can be obtained by using selective culture and coated-dishes respectively which are simple and repeatable methods.

Objective To study the effects of advanced glycation end (AGEs) products induced by bovine serum albumin (BSA) on the survival and the morphology of bovine retinal endothelial cells (BREC) and pericytes (BRP). Methods BSA with the final concentration of 50 mg/ml was incubated in PBS, containing 500 mmol/L D-glucose, for 12 weeks under 37℃. AGEs-BSA was purified by Sephacryl S-300 column chromatography and was confirmed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of AGEs-BSA was determined by the method of commassie protein assay. In order to detect the toxic effects of AGEs-BSA on cultured BREC and BRP, groups of AGEs-BSA and BSA with different concentration and untreated control were set up. Phase contrast microscope was used to observe the effect of AGEs-BSA and BSA (with the concentration of 500 ?g/ml and actuation duration of 48 hours) on morphology of BREC and BRP. Results[WTBZ] As the dosage of AGEs-BSA increased, the number of inhibited cells increased. When the concentration of AGEs-BSA was 500 ?g/ml, the inhibited BREC in AGEs-BSA group was (72.8?15.9)% of which in untreated control group, and the inhibited BRP was (64.8?9) % of which in untreated control group. AGEs-BSA with low concentration promoted the proliferation of endothelial cells, but there was no significant difference between AGEs-BSA and the control group (P=0.231). Inhibited proliferation and abnormal morphology were seen under the phase contrast microscope while the normal morphology of cells was found in BSA and control group. Conclusion AGEs-BSA with the high concentration may inhibit the growth of both BREC and BRP, which leads the loss of BRP and damage of vascular function. These results suggest that nonenzymatic glycosylation plays a major role in diabetic complications.

Objective To investigate if insulin can affect the expression of vascular endothelial growth factor (VEGF) in the retina of streptozotocin-induced diabetic rats. Methods A total of 60 male Sprague-Dawley rats were randomly divided into sodium citrate buffer control group (CIT-CON, n= 30) and STZ-induced diabetic group (STZ-DM, n=30). At the 16th week, 24 rats from CIT-CON group at random were randomly divided to group A (sodium citrate buffer control group, n = 12) and group B (sodium citrate buffer plus insulin group, n= 12). The remaining 6 rats from as CIT-CON group served as negative control. At the same time, 24 rats from STZ-DM group at random were randomly divided to group C (STZ-induced diabetic group, n= 12) and group D (STZ-induced diabetic plus insulin group, n= 12). The remaining 6 rats from STZ-DM group also served as negative control. 4 IU of insulin was injected subcutaneously to rats of group B and D. Immunohistochemistry, Western blot and Real-time polymerase chain reaction (RT-PCR) were used to measure the expression level of VEGF protein and mRNA respectively. RESULTS Insulin significantly increased the VEGF mRNA (7.71 ± 0.25 vs 5.36 ±0. 37, t test P< 0. 05) and protein expression (0. 4925 ± 0. 0122 vs 0. 4272 ± 0. 0110, t test P< 0. 05) in the retina of CIT-CON rats.However, in retina of STZ-DM rats, insulin had no effect on VEGF mRNA (8. 92±0. 27 vs 9. 05±0. 28, t test, P>0. 05) and protein expression (0. 5152±0. 0109 vs 0. 5099±0. 0100, t test P>0.05). Conclusions Insulin had no effect on VEGF expression in the retina of STZ-DM rats.

Objective To research the relationship between quality of life among the elderly with chronic periodontitis (CP) and the severity of CP.Methods A total of 471 elderly patients seeking treatment in the affiliated Stomatology Hospital of Kunming Medical University from September 2013 to January 2014 and their companions were investigated using the questionnaire according to the Chinese version of the Oral health Impact Profile-14.Those who diagnosed with CP were separated into two groups according to Community periodontal index (CPI) through periodontal examination.Group A was diagnosed with the early CP,and Group B was diagnosed with the late CP.Database was established by Epidata and statistical analysis was performed by SPSS.Results A total of 237 elderly patients with early CP and 139 with late CP were recorded.The mean OHIP-14 scores of the former were 14.36 ± 6.67,and the score of the later were 15.65 ± 7.12 relatively.The two groups who lived through negative effects were compared.The results suggested in the 14 items of OHIP-14.The statistical significance existed in the following 4 items,obvious pain appeared,uncomfortable when eating anything,unsatisfied with own diet,become easy to lose one's temper to others (P＜0.05).Conclusion OHR-QOL of elderly patients with the late CP is lower than that of those with the early CP.

Background Most anti-inflammation eyedrops are limited in clinical application owing to multiple adverse effects.A novel peptide GC31 derived from human thrombomodulin has a natural anti-inflammatory activity.Compared with conventional anti-inflammatory eyedrops,GC31 possesses more advantages and potential clinical transforming value.However,relevant study is still lack.Objective The purpose of this study was to evaluate the anti-inflammatory effect of GC31 and the possible mechanisms.Methods Sixty SPF male Wistar rats aged 8-10 weeks were randomized into 6 groups using randomized number table.Non-specific keratitis models were established in 40 rats by intrastromal injection of 10 μl of lipopolysaccharide (LPS) dissolved in PBS.Different doses of GC31 (125 μg or 250 μg) or dexamethason soluble in PBS were sunconjunctically injected in the experimental eyes respectively in the low dose GC31 group,high dose of GC31 group and the dexamethason group,and 10 μl of PBS was used in the same way in the PBS control group.No drug was injected in the model group,and the normal rats were employed as the blank control group.The corneas were examined by slit lamp microscope and were scored based on the criteria of Anand 24 hours after injection.Then the corneas were collected for histopathological examination.Expression of nuclear factor-κB (NF-κB) p65 in the corneas was detected using immunochemistry.Expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) proteins were assayed using ELISA.Real-time PCR was used to detect the expressions of IL-6 mRNA and TNF-α mRNA.The use and care of the experimental animals followed Regulation for the Administration of Affair Concerning Experiment animals by State Science and Techonology Commission.Results A significant difference was seen in the ocular inflammatory scores among the six groups (F =301.238,P =0.000).The inflammatory scores were significantly lower in the high dose of GC31 group than those in the model group (1.85 ± 0.36 versus 2.90± 0.43) (t' =-5.144,P =0.000) ; and the scores in the dexamethason group was lower than those in the high dose of GC31 group(t' =-3.931,P=0.000).Infiltration of inflammatory cells in corneal tissue was milder in the high dose of GC31 and the dexamethason group compared with the model group.The positive response for NF-κB p65 was obviously weaker in the rat corneas in the low and high dose of GC31 groups and the dexamethason group in comparison with the model group.The contents of IL-6 and TNF-α proteins in the corneas were significantly reduced in the low and high dose of GC31 group and the dexamethason group compared with the model group (low dose group:t=-2.626,P=0.009;t'=-2.310,P=0.017.high dose group:t =-3.361,P=0.001 ;t'=-3.151,P=0.002),and the contents of IL-6 and TNF-α proteins in the dexamethason group were lower than those in the high dose of GC31 group (t=-3.361,P=0.001;t'=-3.360,P=0.000).In addition,the expression trend and compared results of IL-6 mRNA and TNF-α mRNA among the groups were similar to those of the IL-6 and TNF-α proteins (all at P＜0.01).Conclusions GC31 suppresses LPS-induced corneal inflammation response by downregulating the expression of inflammatory eytokines.The effect is more dominant in the doses of 250 μg than that in the doses of 125 μg.