BackgroundHypoxia and hyperglycemia are the common causes of retinal neovascularization.In these states,H+ accumulates because of the elevated glycolysis and failure of retinal circulation,thus the retinas readily acidified. ObjectiveThe present study was to explore whether retinal acidosis independently regulates the production of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) and whether the regulation is related to oxidative stress.Methods The retinas from 2-week-old male SD rats were cultured with explant method in DMEM modulated by NaHCO3,and culture retinas were randomly divided into pH 7.2,6.8 and 6.5 groups for 24 hours.In addition,after 24 hours of culture as above described,retinas were washed using PBS two times and then followed by again culture in DMEM with pH 7.2 for another 24 hours.Also,antioxidant was added in different pH values of DMEM for culture as above described.The retinal samples were prepared for histopathological examination.The expressions of VEGF and PEDF proteins and their mRNA in retina tissue were detected by Western blot and fluorescence quantitative polymerase chain reaction (PCR) respectively.Results The retina showed the clear structure and morphology in pH 7.2 group and pH 6.8 group,but retinal vacuoles change was seen in pH 6.5 group after culture for 24 hours.No significant difference was seen in the expressing level of VEGF mRNA in retina between normal group and pH 7.2 group( 112％ ±11％ vs 100％ ±7％ ) (P=0.55),but those in pH 6.8 group and pH 6.5 group were significant increased in comparison with pH 7.2 group( 196％ ±43％ vs 100％ ±7％ ;251％ ±29％vs 100％ ±7％ )( P＜0.05 ).The expressing level of PEDF mRNA in retina in normal group was similar to that of pH7.2 group(86％ ±19％ vs 100％ ±33％) (P=0.64),but that in pH 6.5 group was significantly higher than pH 7.2 group( 230％ ±66％ vs 100％ ±33％ ) ( P＜0.05 ).The resemble results were found in the expressions of VEGF and PEDF protein.After pH reversion,the expressing levels of VEGF mRNA were 100％ ±13％,111％ ±9％,113％ ±9％ in pH 7.2 group,pH 6.8 group and pH 6.5 group respectively without significant difference among them (F=2.51,P=0.16).The expressing levels of PEDF mRNA were 100％ ±13％,110％ ±9％,108％ ±11％in different groups ( F =0.98,P =0.43 ).Under the presence of antioxidant,the expressing level of VEGF mRNA in pH 6.5 group increased in comparison with pH 7.2 group and pH 6.8 group ( P ＜ 0.05 ).The expressing levels of PEDF mRNA were significant different among pH 7.2 group( 100±31 )％,pH 6.8 group( 282±45 )％ and pH 6.5 group(480±117)％ (F=20.73,P=0.00). Conclusions VEGF can be induced by retinal acidification alone,which may be regulated by oxidative stress.Under the retinal acidification,antioxidants promote the expression of PEDF,suggesting that oxidative stress inhibits the production of PEDF.

Child ; Humans ; Plasma Exchange ; Thrombotic Microangiopathies ; therapy

OBJECTIVE To determine the functional role of hydrogen sulfide (H2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca2 +/calmodulin-dependent protein kinaseⅡ (CaMKⅡ) using wild type and CSE knockout mouse models. METHODS Continuous subcutaneous injection isoprenaline (7.5 mg·kg-1·d-1), once a day for 4 weeks to induce heart failure in Male C57BL/6 (6-8 weeks old) mice and CSE-/- mice. 150 μmol·L-1 H2O2 was used to induce oxidative stress in H9c2 cells. Echocardiograph was used to detect cardiac parameters. H&E stain and Masson stain was to observation histopathological changes. Western blot was used to detect protein expression and activity. The siRNA was used to silence protein expression. HPLC was used to detect H2S level. Biotin assay was used to detect the level of S- sulfhydration protein. RESULTS Treatment with S-propyl-L-cysteine (SPRC) or sodium hydrosulfide (NaHS), modulators of blood H2S levels, attenuated the development of heart failure in animals, reduced lipid peroxidation, and preserved mitochondrial function. The inhibition CaMKⅡ phosphorylation by SPRC and NaHS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds. Interestingly, CaMKⅡ activity was found to be elevated in CSE-/- mice as compared to wild type animals and the phosphorylation status of CaMKⅡ appeared to relate to the severity of heart failure. Importantly, in wild type mice SPRC was found to promote S-sulfhydration of CaMKII leading to reduced activity of this protein however, in CSE-/- mice S-sulfhydration was abolished following SPRC treatment. CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of CaMKⅡ is presented. SPRC mediated S-sulfhydration of CaMKII was found to inhibit CaMKⅡ activity and to preserve cardiovascular homeostasis.

OBJECTIVE To determine the functional role of hydrogen sulfide (H2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca2 +/calmodulin-dependent protein kinaseⅡ (CaMKⅡ) using wild type and CSE knockout mouse models. METHODS Continuous subcutaneous injection isoprenaline (7.5 mg·kg-1 per day), once a day for 4 weeks to induce heart failure in male C57BL/6 (6-8 weeks old) mice and CSE-/- mice. 150 μmol·L-1 H2O2 was used to induce oxidative stress in H9c2 cells. Echocardiograph was used to detect cardiac parameters. H&E stain and Masson stain was to observation histopathological changes. Western blot was used to detect protein expression and activity. The siRNA was used to silence protein expression. HPLC was used to detect H2S level. Biotin assay was used to detect the level of S-sulfhydration protein. RESULTS Treatment with S-propyl-L-cysteine (SPRC) or sodium hydrosulfide (NaHS), modulators of blood H2S levels, attenuated the development of heart failure in animals, reduced lipid peroxidation, and preserved mitochondrial function. The inhibition CaMKⅡ phosphorylation by SPRC and NaHS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds. Interestingly, CaMKⅡ activity was found to be elevated in CSE-/- mice as compared to wild type animals and the phosphorylation status of CaMK Ⅱ appeared to relate to the severity of heart failure. Importantly, in wild type mice SPRC was found to promote S-sulfhydration of CaMKⅡ leading to reduced activity of this protein however, in CSE-/- mice S-sulfhydration was abolished following SPRC treatment. CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of CaMKⅡ is presented. SPRC mediated S-sulfhydration of CaMKⅡ was found to inhibit CaMKⅡ activity and to preserve cardiovascular homeostasis.

AIM: To evaluate the efficacy and tolerability of one-site versus two-site phacotrabeculectomy in the treatment of patients with coexisting cataract and glaucoma. METHODS: A comprehensive literature meta-analysis was performed according to the Cochrane Collaboration methodology to identify controlled clinical trials comparing one-site with two-site phacotrabeculectomy. The studies meeting the predefined criteria were reviewed systematically by meta-analysis. Efficacy estimates were measured by standardised mean difference (SMD) for the percentage intraocular pressure (IOP) reduction from baseline to end point, odds ratio (OR) for the percentage having a best-corrected visual acuity (BCVA) of 0.5 or better after surgery and relative risk (RR) for complete success rates. Tolerability estimates were measured by RR for adverse events. All of outcomes were reported with 95% confidence interval (CI). Data were synthesised by Stata 10.1 for Windows. RESULTS: Two-site phacotrabeculectomy was associated with numerically greater, and significant efficacy than one-site in lowering IOP(SMD,-0.19;95% CI, -0.33 to -0.04; P=0.01). Numerically greater, but nonsignificant proportions of two-site patients than one-site patients had a BCVA of 0.5 or better (OR, 0.65; 95% CI, 0.30 to 1.39; P=0.26).Numerically greater, but nonsignificant proportions of two-site patients than one-site patients achieved the target IOP without anti-glaucoma medication at the end point (RR, 0.94; 95% CI, 0.84 to 1.04; P=0.22). Furthermore, there was no significant difference in adverse events between two surgical procedures.CONCLUSION: The efficacy of two-site phacotrabeculectomy appears to be superior to one-site phacotrabeculectomy. One-site and two-site phacotrabeculectomy are similarly tolerable in postoperative adverse events.

AIM:To compare one-site vs two-site phacotrabe-culectomy in chronic angle-closure glaucoma (CACG) coexisting with cataract.METHODS:This prospective, randomized study included 41 eyes with CACG. One-site approach was performed in 21 eyes and two-site procedure in 20 eyes. Intraocular pressure (IOP), best-corrected visual acuity (BCVA), the number of antiglaucoma medications and complications were observed. All patients were followed up for 9 months.RESULTS:There were no significant differences between the two groups preoperatively. IOP decreased from 22.7±4.9mmHg and 23.7±4.7mmHg preoperatively in one-and two-site groups to 18.0±1.2mmHg and 16.7±1.1mmHg 9 months after operation respectively(P<0.05). There were no significant differences in mean IOP between the two groups at any time (P>0.05). Decrease of the number of antiglaucoma medications and BCVA improvement were similar in both groups 9 months after surgery (P>0.05).There were no significant differences in complications between the two surgical procedures.CONCLUSION:There were no significant differences between the two groups in clinical efficacy and complications.

Objective To explore the expression and mutual relationship between transforming growth factor-?(TGF-?) and hepatocyte growth factor(HGF) in children's renal tissues,as well as the effect of the 2 indexes on the renal pathological change.Methods According to the severity of renal pathological change under light microscope,61 cases were divided into 3 groups:named control group(26 cases,clinically diagnosed as thin basement membrane nephropathy),test group Ⅰ [22 cases,clinically diagnosed as less evident focal segmental glomerulosclerosi(FSGS) nephropathy] and test group Ⅱ(13 cases,clinically diagnosed as evident FSGS nephropathy).Immunity class test(SP method:streptavidinbiotin peroxidase method) was used to detect the representation of HGF and TGF-?.Semi-quantitative analysis had been carried out in all cases.Film reading of cell was viewed by Olympus microscope,brown yellow from cytoplase as the positive signal,10 high power microscope visions were randomly selected from renal glomerali area and 10 from renal interstitium area.Medical image analysis software was used to determine the masculine area of HGF or TGF-? and image intensity;then the immunity class index was defined as masculine area ? image intensity.Results 1.HGF and TGF-? existed in all renal tissues;2.Expressions of HGF and TGF-? increased obviously along with FSGS pathology alteration(Pa

Background Antagonists against vascular endothelial growth factor (VECF) play key roles in treating and preventing neovascular ophthalmopathy. As a novel anti-angiogenic factor, insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) might be an antagonist against VEGF in eye. Objective This study was to explore the inhibitory effect of IGFBP-rP1, a novel anti-angiogenic factor, on VEGF-induced retinal angiogenesis in vitro. Methods The retina-choroid endothelial cell line ( RF/6A ) was cultured in DMEM containing 10% fetal bovine serum. Culture cells were divided into control group(free-serum culture group) ,10mg/L VEGF culture group and different concentrations of IGFBP-rP1 (50,100,200 mg/L) +10 mg/L VEGF group. The expression of IGFBP-rP1 in the cells was detected by immunofluorescence assay. The proliferation and migration of RF/6A cells were evaluated using MTS colorimetric assay and the chemotactic motility assay, respectively. Flow cytometry was used to detect the apoptosis of RF/6A cells. Results The immunofluorescence assay RF/6A cells showed the green fluorescence in cytoplasm and red fluorescence in nuclei after cells were exposed to any concentration of IGFBP-rP1 ,but only red fluorescence was seen in nuclei in control cells. After stimulation of 10 mg/L VEGF,the proliferation value (A490) was elevated and the numbers of cell migration were increased in comparison with control group (t = -15. 191, P = 0. 000; t = -21. 274, P = 0. 000 ) , but the cellular apoptosis rate was lower than the control group (t - 10. 228, P = 0. 000 ) . After treated with various concentrations of IGFBP-rP +10% VEGF, the proliferation and migration of RF/6A cells were significantly decreased in comparison with only 10% VEGF group (F = 534. 158,P = 0. 000;F = 2742. 323,P = 0.000,respectively) ,and the inhibitory effects were gradually enhanced with the increase of IGFBP-rP1 levels (P<0. 05). The apoptosis rate of RF/6A cells in 50,100 and 200 mg/L + 10 mg/L VEGF groups increased by ( 1. 26±0. 04)% ,( 1. 50±0. 07)% and ( 1. 93±0. 27)% respectively,showing significant differences among different groups ( F = 274. 273, P = 0. 000). Conclusion IGFBP-rP1 inhibits the proliferation and activity of retina and choroid endothelial cells induced by VEGF at a concentration-independent manner. It appears to be as a novel endogenous inhibitory factor in retinal angiogenesis.

DNA microarrays have been acknowledged to represent a promising approach for the detection of viral pathogens. However, the probes designed for current arrays could cover only part of the given viral variants, that could result in false-negative or ambiguous data. If all the variants are to be covered, the requirement for more probes would render much higher spot density and thus higher cost of the arrays. Here we have developed a new strategy for oligonucleotide probe design. Using type I human immunodeficiency virus (HIV-1) tat gene as an example, we designed the array probes and validated the optimized parameters in silico. Results show that the oligo number is significantly reduced comparing with the existing methods, while specificity and hybridization efficiency remain intact. The adoption of this method in reducing the oligo numbers could increase the detection capacity for DNA microarrays, and would significantly lower the manufacturing cost for making array chips.

BACKGROUNDPhacotrabeculectomy can be performed using one-site or two-site incisions. This meta-analysis evaluated the efficacy and tolerability of one-site versus two-site phacotrabeculectomy in the treatment of patients with coexisting cataract and glaucoma.

METHODSA comprehensive literature search was performed according to the Cochrane Collaboration methodology to identify randomized controlled clinical trials comparing one-site with two-site phacotrabeculectomy. Studies meeting our predefined criteria were included in the meta-analysis. Efficacy estimates were measured by weighted mean difference (WMD) for the percentage intraocular pressure (IOP) reduction from baseline to end point, relative risk (RR) for the proportion of patients with a best-corrected visual acuity (BCVA) of 0.5 or better after surgery and complete success rates. Tolerability estimates were measured by RR for adverse events. All of outcomes were reported with 95% confidence interval (95%CI). Data were synthesised by Stata 10.1 for Windows.

RESULTSTwo-site phacotrabeculectomy was associated with greater reductions in IOP than the one-site procedure (WMD: -5.99, 95%CI: -10.74 - -1.24, P = 0.01). A greater proportion of patients also achieved a BCVA of 0.5 or better (RR: 0.91, 95%CI: 0.74 - 1.12, P = 0.36) and the target IOP without anti-glaucoma medication at the study end point (RR: 0.94, 95%CI: 0.83 - 1.07, P = 0.34) after two-site than one-site phacotrabeculectomy, but the differences were not significant. There were no significant differences in adverse events between two surgical procedures.

CONCLUSIONSTwo-site phacotrabeculectomy is superior to one-site phacotrabeculectomy in reducing IOP, but other post-operative effects are similar. One-site and two-site phacotrabeculectomies have similar adverse event rates.