Background Glucocorticoid drugs have been used increasingly in ophthalmology.It has been established that glucocorticoid are associated with a rise in intraocular pressure (IOP) and the development of glaucoma.Selective laser trabeculoplasty (SLT) is thought to be effective in treating patients with glucocorticoidinduced elevated IOP.Objective This study was to assess the efficacy of SLT in lowering IOP in patients with glucocorticoid-induced ocular hypertension.Methods A retrospective case series study was adopted.SLT around 360° chamber angle was performed in 9 eyes of 9 patients with glucocorticoid-induced glaucoma in Eye & ENT Hospital of Fudan University,including 5 eyes of 5 patients owing to use of glucocorticoid eye drops for long-term after laser in situ keratomileusis (LASIK) and 4 eyes of 4 patients who received intravitreal injection of 0.1 ml triamcinolone acetonide (TA) (4.0 mg) for macular edema induced by central retinal vein occlusion(CRVO).All of the patients were lack of preexisting glaucoma or ocular hypertension and underwent unsuccessful maximum tolerated medical therapy before SLT treatment.The base IOP was 35-44 mmHg and glucocorticoid drugs were ceased for 2-12 months prior to the SLT.The patients were followed-up for 6 months.IOP was measured and recorded before and 1 hour,1 week,1 month and 3 months,6 months after SLT.The difference of IOP was compared by repeated measures analysis of variance and multiple comparison analysis.Results All the patients received single SLT operation.The mean IOP was (40.0±2.9) mmHg before operation,but the IOP was (37.9±8.1),(34.9±5.9),(27.6±6.7),(21.6±6.9) and (17.9±2.9)mmHg 1 hour,1 week,1 month,3 months and 6 months after SLT.The IOP at 1 month,3 months and 6 months after SLT were significantly lower than that before operation (all at P＜0.05).Two patients received a filtration surgery for uncontrolled IOP at 1 month and 3 months after SLT respectively,and another patient still used 2 kinds of lowing-IOP eye drops until the end of following-up duration.Conclusions SLT reduce IOP in 6 eyes of 9 patients with glucocorticoid-induced increased IOP from the initial 1 month through 6 months after SLT.

Objective To investigate the protective effects of low-molecular heparin and urokinase on glomerular inflammation. Methods Forty 3-month-old female Wistar rats were randomly divided into five groups with 8 rats in each group: normal control (NC) group; lipopolysaccharide (LPS) group; LPS and tranexamic acid (LT) group; LPS, tranexamic acid and low-molecular heparin (LTH) group; and LPS, tranexamic acid and urokinase (LTU) group. Fibrin deposition and CD11b positive cells were identified by immunohistochemical staining. Western blotting was used to determine the ICAM-1 expression. Results No fibrin depositions and CD11b positive cells were found in glomeruli of rats in NC group. Compared with NC group, fibrin deposit (9.1%?1.6%), CD11b positive cell (11.2?2.1) and ICAM-1 expressions (0.23?0.09) were significantly increased in L group (P0.05). Conclusion Both low-molecular heparin and urokinase can effectively decrease fibrin deposits and alleviate inflammation.

Objective To prepare mouse polyclonal antibody against human Nanog by genetic immunization and to identify this antibody by Western blot and immunofluorescence. Method The antigenicity fragment (A16-V101) of human Nanog (hNanog) was chosen by analysis of Accelrys software, and its cDNA (258bp) was amplified from plasmid containing full-length cDNA of hNanog, then it was cloned into pBQAP-TT to construct recombinant plasmid pBQAP-TT-hNanog for genetic immunization. Mice were immunized with this recombinant plasmid and two other adjuvant plasmids-pCMVi-GMCSF and pCMVi-FIT3L, which help to enhance the antibody's generation. After 12 weeks, we obtained mouse anti-hNanog antibody from mice blood serum. The antibody titer was determined by enzyme-linked immunoadsordent assay (ELISA), and its specificity was identified by Western blot in human renal protein. Using this antibody, we detected hNanog expression in HKC cells of hNanog-AAV2 transfection. Results Recombinant plasmid pBQAP-TT-hNanog for genetic immunization was confirmed to be correct by restriction digestion and sequencing. The result of ELISA showed that the antibody titer was 1∶3 200. This antibody recognized a band of 34kD hNanog protein in human renal protein by Western blot. Immunofluorescence showed that Nanog protein was mainly located in the nuclei in hNanog transgene HKC cells. Conclusion Genetic immunization can offer mouse anti-hNanog polyclonal antibody of high titer and high specificity.

Objective To explore the changes of E-cadherin expression in renal ischemia-reperfusion injury.Methods For the in vitro analysis of epithelial ischemia,confluent monolayers of MDCK cells growing in DMEM were depleted of ATP for 4 h by incubation in PBS (supplemented with 1.5 mM CaCl2 and 2 mM MgCl2) containing 10 ?M antimycinA.For the in vivo studies of epithelial ischemic injury,adult Sprague-Dawley rats were subjected to bilateral renal artery ligation.Renal pathological changes were measured by PAS stain.Location and expression of E-cadherin were detected by immunohistochemistry and western blot respectively.Results E-cadherin were primarily found in a linear pattern at the lateral portions of the plasma membrane in normal MDCK.After ATP depletion for 4 hours,the linear pattern altered and manifested by the appearance of intracellular staining.In invivo ischemia-reperfusion model rats,E-cadherin expression was changed from normal tubular epithelial cell basal membrane to cytoplasma.Western blot suggested that in sham-operated rats,E-cadherin was 120 ku lane vs 80 ku lane in ischemia for 60 min rats,while in ischemia for 45 min rats,both the 120 ku and 80 ku lanes were detected.Conclusion In renal ischemia-reperfusion,the location and expression of E-cadherin are obviously altered in vivo and in vitro study and E-cadherin are degradated as ischemia time prolongs.These changes may be the reason why tubular epithelial cell exfoliated from TBM in ischemia-reperfusion injury.

Objective To analyze mRNA and protein expressions of plasminogen activator and plasminogen activator inhibitor-1 using microdissection and quantitative real-time PCR,and to analyse their associations with microvascular diseases in IgA nephropathy.Methods Twenty-four IgA nephropathy patients treated in Kidney Center and Kidney Laboratory of PLA from 2000 to 2004 were admitted into the study;20 renal glomeruli or 200 renal tubules and peritubular interstitial per patient were captured by Laser Microdissection System from IgA nephropathy renal biopsy slides(8 ?m),and mRNA was extracted.PA mRNA and PAI-1 mRNA levels in glomerulus and tubulointerstitial area were measured by Taqman quantitative real-time PCR.Capillaries densities in glomerulus and tubulointerstitial area were measured using immunohistological staining method and computer image analysis system.Results With the aggravation of lesions,the glomeruli and peritubular capillaries densities of IgA nephropathy decreased.The glomerular and peritubular capillaries densities were negatively associated with the level of serum creatinine(R2=0.6946 and R2=0.6271,P

Objective To investigate the characteristics and proper use of anticoagulants in hemodialysis (HD). Methods Thirty-one HD patients were enrolled in the study. Unfractionated heparins (UFH), dalteparin sodium or argatroban were used for HD anticoagulation respectively. Blood specimens were taken from the arterial line at the beginning (0 h) and at the end of HD (4 h), and from the arterial (2a) and the venous (2v) line respectively at 2 h of the HD session. Glass bead activated clotting time (gbACT), clot rate (CR) and platelet function (PF) were examined by Sonoclot analyzer. Prothrombin fragment 1+2 (PF1+2) and granule membrane protein-140 (GMP-140) were assayed by ELISA. Meanwhile, blood was taken from 8 healthy volunteers to examine the above parameters as control. Results (1) Compared with the control group, CR, PF1+2, PF, GMP-140 were increased significantly in all the patients (P<0.05). (2) UFH group:Compared with the 0 h point, gbACT of other time points increased significantly (P<0.05), CR, PF, and PF1+2 decreased significantly (P<0.05). Compared with the control group, gbACT increased (P<0.05) and CR decreased (P<0.05) significantly at the end of the sessions. (3) Daheparin sodium group: Compared with the 0 h point, gbACT of 2a point increased significantly (P<0.05), CR and PF1+2 of 2a, 2v and 4 h points decreased significantly (P<0.05), meanwhile, the extents of increased gbACT and decreased CR from the arterial line were greater than those from the venous line. Compared with the control group, gbACT increased significantly at the end of HD session (P<0.05), but CR was not significantly different. (4) Argatroban group: There were no significant differences of gbACT between 0 h and other time points. CR of 2a, 2v points decreased obviously than that of 0 h point, and CR of 2v decreased more significantly. CR of 2a point was not different from the control group, while CR of 4 h point was greater as compared to control group. During the monitoring, PF1 +2 tended to increase. Conclusions With intensive anticoagulant effect, UFH may induce the risk of hemorrhage not only during but also after the dialysis sessions. Dalteparin sodium, a good anticoagulant, is stir related with the risk of hemorrhage during HD. Argatroban is an ideal anticoagulant for patients with the risk of hemorrhage.

Objective To explore the clinicopathological features of IgA nephrolpathy associated with malignant hypertension (IgAN-MHT) and to analyze their correlation with renal vascular lesions. Methods Twenty-nine patients of IgAN-MHT were screened from 2000 biopsy-proven eases with primary IgA nephropathy (IgAN) in our department from April 1997 to May 2007. Data of clinicopathology and follow-up of these 29 patients were collected. Semi- quantitative analysis was performed to evaluate the pathological changes. Inner lumen, outer lumen, intimal thickness, tunica media-to-internal lumen ratio of 436 arterioles, 124 interlobular arteries and 5 arcuate arteries were measured. The primary endpeint was the composite of a doubling of serum creatinine level and ESRD. Correlations of renal vascular lesions with clinical manifestation, pathological change and prognosis were examined by Spearman and Cox methods. Results 1.5% of all the IgAN patients presented malignant hypertension. The common clinical features were renal failure (100%), hyperurieacidemia (62.7%) and hypertriglyceridemia (51.7%). The average amount of urine protein excretion was 2.8 g/d. The common pathological changes were moderate mesangial proliferation, severe global sclerosis, severe interstitial inflammation and severe interstitial- tubular fibrosis. The small arteries (arcuate arteries and interlobular arteries) and arterioles (afferent arterioles) were both involved in IgAN-MHT. The characteristic lesions of intrarenal arteries included vascular occlusion, media thickening, proliferative endarteritis (onionskin lesion, musculomucoid intimal hyperplasia), hyaline arteriosclerosis, but mainly vascular occlusion (86.2%). The arteriole lesion was negatively correlated with age and total protein level; vascular occlusion was positively correlated with uric acid level. The average foUow-up period was 21.1 months. Forteen patients reached the endpoint. The arteriole lesion was the main independent risk factor for the progression of IgAN-MHT (RR=10.21, 95%CI=1.16～89.67). Conclusions The main clinical feature of IgAN-MHT is renal failure. The main histological feature of intrarenal vascular lesions is occludes arterioles. Arteriole lesion is the main independent risk factor for the progression of IgAN-MHT.

This study is designed to investigate the effects of chinfloxacin hydrochloride (CFX) on the kinetics of HERG K+ channel. Whole cell patch clamp technique was used to record HERG K+ currents from HEK293 cells transiently transfected with cgi-HERG-GFP plasmids and channel kinetics were assessed in the absence and presence of CFX and moxifloxacin hydrochloride (MOX). Results demonstrated that the open state of HERG K+ channel was inhibited by CFX in a concentration- and time-dependent manner, with an IC50 of 162.1 +/- 14.2 micromol x L(-1), two folds higher than its positive control MOX. But there were no significant effects on channel kinetics. In addition, the inhibitory effect of CFX on HERG was enhanced when cells were subjected to altered extracellular K+ concentration.

Objective:To prepare anti-human IgE nanobody by phage display technology,and to establish a method for hyper-sensitivity detection of cat dander specific IgE antibody.Methods:Allergen bio-information of cat was searched in WHO/IUIS Allergen Database.After synthesizing sequence,recombinant cat dander allergenic protein Fel d 1 was expressed and purified in prokaryotic ex-pression system.Human IgE was used to immunize Bactrian camel and RNA were extracted from lymphocyte to construct phage dis-play library.Library capacity,diversity and insertion rate were analyzed,anti-human IgE nanobody were obtained by screening and identification.A magnetic particle chemical method for cat dander specific IgE antibody detection was established using recombinant allergen-coupled magnetic particles and acridine ester-labeled nanobodies.Results:Capacity of phage display library was 1.88×108 cfu/ml,insertion rate was 93.6%,and purity of nanobody was>95%.Linear range of the method based on nanobody was 0.1～100 U/ml,who was consistent with ImmunoCAP detection system by clinical data.Conclusion:Nanobody-based cat dander specific IgE antibody hypersensitivity assay is successfully prepared,providing a technical basis for auxiliary diagnosis of cat allergic diseases.

Histone acetylation is one of the most important reactions of post-translational modification of histones, which plays an important role in the regulation of epigenetic processes. Histone deacetylase 2 as a member of type I histone deacetylases,involved in the catalytic regulation of histone and a variety of non-histone deacetylation,regulates a variety of life processes. This paper summarizes the basic structure of histone deacetylase 2 and the role of histone deacetylase 2 in various diseases,and provides a theoretical basis for conducting related studies.