Background Trans-arachidonic acids (TAAs) are specific lipid markers of nitrative stress and the major products of NO2·-mediated ismoerization of aracchidonic acid(AA),and they represent a possible new mechanism of NO2·-induced toxicity in ischemic retinopathy.Our previous study demonstrated that high glucose induce nitrative stress response. Objective This study aimed to evaluate the effect of nitrative stress on retinal capillary in diabetic rats by detecting the TAAs with gas chromatography and mass spectrometry(GC/MS).Methods One hundred 2-week-old clean male SD rats were randomly divided into control group and diabetic group.Diabetic rat models were induced by intraperitoneal injection of streptozoticin of 60 mg/kg,and the successful models were identified as the blood glucose level>16.7 mmol/L.The synthesis method was modified in this study andaraehidonic acid w8$utilized as starting material.14E-AA was obtained as a standard sample via expoxidation and deoxidation in a safe and practical pattern.Serum TAAs and AA in diabetic rats were detected by GC/MS in 2,4,8,12 and 16 weeks after injection of streptozoticin.The ratio of peak area of TAAs to AA with selected ion of 79 was estimated and compared with normal rats. Results Fifty rats had been in Diabetic models were established in 50 rats with the successful rate 100%,and the high blood glucose level in model rats retained throughout the experimental duration.GC/MS results showed that the 14E-AA was obtained with the purity of 97.3%,which provided a standard sample to nitrative stress-related research as a premise. No significant differences were found in serum TAAs/AA values at 2 weeks and 4 weeks between model group and control group (t =-0. 376, t =-0. 642,P>0. 05). However,serum TAAs/AA values in at 8,12 and 16 weeks after streptozoticin injection were considerably elevated in comparison with those of control group (0. 0832 vs 0. 1042,0. 0910 vs 0. 1568, 0. 1100 vs 0. 1895;t=-36.409,t=-166.714,t=-148.212,P<0. 05). Conclusion Elevation of serum TAAs/AA is associated with diabetic duration,implying that damage of nitrative stress to retinal capillary is a parallel procedure to diabetic course.Establishment of GC/MS detection system offers a new evaluating indicator in the research on microvascular ischemic disease and could be implemented in clinical testing.

Objective To assess the effect of case-based learning (CBL) in ophthalmology teaching.Methods Lecture based learning combined with CBL was applied in ophthalmology teaching for 19 undergraduates majored in clinical medicine of 2007 grade from Shanghai Jiao Tong University School of Medicine.Questionnaire survey was conducted for students after the course ended to understand their evaluation on CBL.Proportion of selected data was analyzed comparatively.Results 100％ (19/19) students believed that CBL was essential in ophthalmology teaching and rational at the end of course.84.2％ (16/19) students accounted that CBL could only be assistant teaching method.94.7％ (18/19) and 84.2％ (16/19) students considered that CBL was beneficial for cognition of eye diseases completely and clinical thinking formation.4-6 class hours for CBL were approved by 73.7％ (14/19) students.Conclusions CBL is essential in ophthalmology teaching and is accepted and supported by students,therefore it should be the main assistant teaching method.

Sailuotong (SLT) is a compound preparation composed of ginseng, ginkgo and saffron for the treatment of vascular dementia In order to identify its material foundation and provide evidence for therapeutic regimen, the pharmacokinetics and brain distribution of ginsenosides were investigated after intragastric administration of SLT. An LC-MS/MS method was developed for the determination of 7 ginsenosides in rat plasma simultaneously. Statistical analysis of obtained data demonstrated that the method has achieved the desired linearity, precision, accuracy and sensitivity. After administration of SLT at 60 mg x kg(-1) dose, 7 ginsengosides were all absorbed into systematic circulation. The quantitative and statistical analysis of gensenosides in plasma showed that protopanaxdiol saponins exhibited higher concentration and longer half life than protopanaxatriol saponins. The mean value of half life of ginsenosides Rg1, Re, Rb1, Rb2/b3, Rc and Rd were 15.26, 2.46, 18.41, 27.70, 21.86 and 61.58 h respectively. The peak concentration of them were 7.15, 2.83, 55.32, 30.22, 21.42, 8.81 microg x L(-1) respectively. The determination of brain distribution at different time after dosing revealed ginsenosides entered into brain promptly but the concentration declined along with time rapidly. The ginsenosides with higher concentration in brain were Rg1, Re, Rb1 and Rc. These findings demonstrated ginsenosides could be absorbed in blood and penetrated into brain rapidly. Some ginsenosides, especially Rg1 and Re, might be the main components directly effecting neurocyte in brain taking advantage of their better brain distribution. While ginsenosides of mostly protopanaxdiol saponins might protect brain mainly depending on peripheral efficacy in virtue of their long residence in blood, by which higher concentration could be reached after multiple dosing.
Animals ; Brain ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Ginsenosides ; pharmacokinetics ; Male ; Rats ; Rats, Wistar ; Time Factors

Unicellular green alga,Dunaliella salina(D.salina),is a biflagellar alga without cell wall,which is a kind of very important eukaryotic microalga.In the previous study,the research of D.salina focus on the morphology,the mechanism of salt tolerance and ?-carotene,however,with the rapid development of microalgal biotechnology,a lot of work about D.salina was reported in recent years.In the area of molecular biology,the studies of D.salina mainly place emphasis on the cloning and analysis of important functional genes,regulatory sequences,and the expression of foreign genes using D.salina as host.The research advance in these aspects were reviewed.

The human canstatin cDNA was amplified by RT-PCR and then directionally cloned into pU? expression vector. The recombinant pU?-Can vector was connected with the screening marker (bar box), to construct a eukaryotic expression vector called pU?-Can-Bar. This expression vector was introduced into the D.salina by glass beads method. The screening culture of transformants of D.salina was performed in solid media containing 5 ?g/ml PPT, and the analyses of transformants were carried out through PCR and Southern blot. PCR results revealed that specific 700 bp products were detected in the different transformants of D.salina but not in negative control. Southern blot analysis further demonstrated that human canstatin gene was integrated into the D.salina genome. Moreover, the results of genetic stability analyses of transformants demonstrated that canstatin gene was stably inherited in the D.salina transformants. The successful preparation of the D.salina transformants will provide the experimentation evidence for producing canstatin protein cosmically by using the D.salina bioreactor and give a better prophase work basis for clinic application of canstatin protein early.

BACKGROUND:Stem cel s can be induced to differentiate into dopaminergic neurons in vivo and in vitro, which provides a theoretical basis for stem cel transplantation in the treatment of Parkinson’s disease
OBJECTIVE:To explore the feasibility and mechanism of intracerebral transplantation of umbilical cord blood mesenchymal stem cel s for treatment of Parkinson’s disease rats.
METHODS:Intracerebral injection of 6-hydroxydopamine was used to make Parkinson’s disease models in SD rats. Twenty-two model rats were randomized into cel transplantation group (n=12) and control group (n=10) and respectively injected intracerebral y with umbilical cord blood mesenchymal stem cel suspension and PBS. At 1-8 weeks after cel transplantation, intra-abdominal injection of apomorphine was performed every week to observe the rotation behaviors of rats;at the 2nd and 8th weeks, rat’s striatum and substantia nigra were taken for immunohistochemistry staining.
RESULTS AND CONCLUSION:The rotation behaviors were gradual y decreased with time in the cel transplantation, but had no changes in the control group. At 3-8 weeks after transplantation, there were significant differences in the rotation behaviors between the two groups (P<0.05). At 2 weeks after transplantation, tyrosine hydroxylase-positive cel s were found within and around the striatum of the cel transplantation group;but there were no exogenous cel s in the control group. At 8 weeks after transplantation, there were stil active cel s and tyrosine hydroxylase-positive cel s in the striatum of cel transplantation group, and there was no tyrosine hydroxylase expression in the striatum of the control group. These findings suggest that transplanted umbilical cord blood mesenchymal stem cel s can survive in the brain that are positive for tyrosine hydroxylase, which can improve the behavior abnormalities of Parkinson’s disease rats.

Objective To analyze and compare the clinical and laboratory features between patients with ankylosing spondylitis (AS) and nonradiographic axial spondyloarthritis (nr-axSpA).Methods One hundred and forty-one patients with AS and 73 cases with nr-axSpA were recruited.Clinical and laboratory indexes of individuals were recorded in detail,Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Ankylosing Spondylitis Disease Activity Score (ASDAS) crp were calculated.Spondyloarthritis research consortium of Canada (SPARCC) score standard was used to evaluate the degree of bone marrow edema in sacr-oiliac joint under magnetic resonance imaging scanning.T test,rank test and x2 test were used for statistical analysis.Results The average age of patients with AS was obviously higher than that of patients with nr-axSpA (t=4.962,P＜0.01).Patients with AS were more often male,and those with nr-axSpA were more often female (82.0％ of the AS patients were men and 49.3％ of the nr-axSpA patients were men (x2=24.079,P＜0.01).Disease duration of AS was significantly longer than that of the nr-axSpA (Z=6.396,P＜0.01).The incidence of human leukocyte antigen (HLA)-B27 positive in AS was 89.4％,which was similar to that in patients with nr-axSpA (84.9％) (x2=0.884,P＞0.05).21.6％ (21 cases) of patients with AS had peripheral swollen joints,which was higher than that in nr-axSpA (2.2％,x2=8.861,P=0.003).Forty cases in AS had tender joints (41.2％),only 6 patients in nr-axSpA had tender joints (13.3％,x2=11.458,P＜0.01).Serum erythrocyte sedimentation rate (ESR) and C reactive protein (CRP) levels in patients with AS were clearly higher than those in nr-axSpA (P＜0.01) patients.In AS,the patient global assessment (PGA),BASDAI and ASDAScrp value was significantly higher than that in nr-axSpA (P＜0.01).There were no differences in SPARCC score or incidence of bone marrow edema in sacroiliac joint in magnetic resonance imaging scanning between AS and nr-axSpA (P＞0.05).Percentage of patients with occipit-to-wall distance higher than 0 cm in AS was higher than that in nr-axSpA,and the mean distance of fingers to ground in AS was also higher than that in nr-axSpA (x2=19.844,P＜0.01;Z=5.724,P＜0.01).Chest expansion degree and Schboer's test in AS was much lower than that in nr-axSpA,respectively (Z=3.083,P=0.002;Z=5.103,P＜0.01).BASFI in AS was higher than that in nr-axSpA (Z=5.840,P＜0.01).The ratio of joint function in AS was obviously worse than that in nr-axSpA (x2=1 1.369,P=0.01).Conclusion Compared to patients with nr-axSpA,AS patients are male predominant,and have severer inflammation in clinical and laboratory findings and are worse in functional status.

AIM: To investigate the effects of Astragalus polysacharin(APS) on human fibroblast and human umbilical vein endothelia cell (HUVEC) proliferation, as well as its acts on adhesion between white cells and HUVECs. METHODS: Human fibroblasts from distal and proximal skin away the ulcer were cultured as normal fibroblasts(NF) and wounded fibroblasts(WF). MTT assay was used for detecting cell proliferation, Rose Bengal staining and fluorescence immunohistology assay were used for examining the adhesion of human polymorpho-nuclear cell(PMN) and TPH-1 to HUVECs. RESULTS: 2 44-156 mg/L APS promoted WF proliferation, and 2 44-39 mg/L APS also promoted NF proliferation, but it did not show any proliferating effect on HUVECs. APS inhibited the adhesion of PMN or TPH-1 to HUVECs induced by tumor necrosis factor(TNF). At 25-100 mg/L, it also inhibited both VCAM-1 and ICAM-1 expression in HUVECs induced by TNF. Treatment with APS for 12 h also inhibited CD44 expression in HUVECs. CONCLUSION: APS shows mitogenic activity on both human normal and wounded fibroblasts. It also exerts anti-inflammation effects by inhibiting adhesion molecule expression and adhesion of white cells to HUVECs. [

Objective To investigate the role of microRNA-129-5p (miR-129-5p) in the regulation of epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs) isolated from peritoneal dialysate effluents and TGF-β1 induced HPMCs line.Methods The isolated cells were cultured from peritoneal dialysate effluents overnight of 10 patients just started PD and 12 patients with PD over 6 months.Taqman PCR assay was used to determine the expression of miR-129-5p in the HPMCs.Moreover,the expression of miR-129-5p in HPMCs induced by 5 μg/L TGF-β1 for 0-72 h was also detected by Taqman PCR.HPMCs were pre-transfected with miR-129-5p precursor (pre-mir-129-5p) to overexpress miR-129-5p,then incubated with TGF-β1 for 48 h,and the expression of EMT associated gene and protein was detected by real-time PCR,Western blotting and immunofluorescence,respectively.Furthermore,the effect of TGF-β1 on the expression of Smad interacting protein-1 (SIP1) and the regulation of pre-miR-129-5p on the SIP1 expression also were investigated.Results MiR-129-5p expression significantly down-regulated in the HPMCs isolated from PD patients over 6 months than from PD start patients(P ＜ 0.01).Similarly,TGF-β1 remarkably decreased miR-129-5p in HPMCs lines on time-dependent manner (P ＜ 0.01).Pre-mir-129-5p dramatically restored the expression of epithelial marker E-cadherin,while inhibited the expression of Vimentin,a mesenchymal marker,in HPMCs induced by TGF-β1 (all P ＜ 0.01).In addition,TGF-β1 increased SIP1 expression in HPMCs time dependently,while the high level of SIP1 protein was obviously repressed after transfected of pre-miR-129-5p (P ＜ 0.01),but there was no obvious change of its mRNA expression.Conclusion MiR-129-5p modulates EMT formation of HPMCs in PD process,possibly by posttranscriptional inhibition of SIP1.Targeting miR-129-5p/SIP1 may provide a new approach for the prevention and treatment of peritoneal fibrosis during PD.

ObjectiveTo evaluate the applicability of modified formulas based on plasma creatinine levels in Chinese patients with chronic kidney disease (CKD). MethodsA total of 327 CKD patients were investigated. Glomerular filtration rate (GFR) was estimated with Chinese equations and Ruijin equation. The accuracy of estimated GFR was compared with 99mTc-DTPA-GFR (sGFR) in CKD patients. ResultsBland-Ahman analysis demonstrated that Ruijin equation was more consistent with sGFR than the other equations. But all the equations were not well consistent with sGFR. Linear regression showed that the slopes of Ruijin equation and MDRD-1 equation were closer to the identical line. 15%, 30% and 50% accuracy of Ruijin equation were higher than the other equations. But 30% accuracy of Ruijin equation was still less than 70%. When the accuracy of estimated GFRs was compared with sGFR in different stages of CKD, GFR estimated by Ruijin equation showed good results. ConclusionsWhen plasma creatinine is checked with enzymatic method, modified GFR estimation equations may show great bias in Chinese CKD patients. More clinical trails should be carried out to evaluate and identify the application of modified GFR estimation equations in Chinese patients with CKD.