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.

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

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.

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 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.

AIM:To study the anti-tumor effect and mechanism of fusion vaccine on DCs and C6 glioma cells.METHODS: PEG was used to fuse DCs with C6 glioma cells.Immunofluorescence with GFAP-FITC was used to identify the DC/C6 fusion cells.Rat brain glioma models were made by stereotactic technique.After 5 days of inoculation of C6,107 fusion cells were injected through tail vein in group A.The same number of DCs and the same volume of PBS were used in group B and group C.The survival time of rats in these three groups was analyzed by Log-rank survival analysis.Tumor samples were checked by HE staining and immunohistochemical staining with CD8Mcab.RESULTS: Positive result of GFAP-FITC immunofluorescence was observed in DC/C6 fusion cells.The Log-rank survival analysis showed that statistically significant difference in group A was observed compared to that in group B and group C(P

Objective:To analyze the prevalence of epilepsy and its clinical features in patients with Alzheimer's disease(AD).Methods:Clinical data of patients with AD from a separate database of Tianjin Huanhu Hospital for dementia and degenerative diseases from January 2010 to January 2019 were retrospectively examined.The prevalence of AD combined with epilepsy was analyzed.The characteristics from neuropsychological assessment, clinical manifestations of epileptic attacks, electroencephalogic patterns and other features were analyzed and compared with those of AD patients without epilepsy.Results:Of 376 AD patients, 11.4% had epilepsy, with 4.0% showing non-induced seizures before AD diagnosis and 7.5% after AD diagnosis.The average age of AD patients at first onset of epilepsy was 59.8±19.9 years, and the average age of patients at initial diagnosis with AD was 62.8±6.6 years.The most common type was focal seizures(65.1%), and EEG showed mostly focal slow waves(69.8%)and focal epileptiform discharges(23.3%). The AD diagnosis for patients with epilepsy was 8 years earlier than those without epilepsy.Conclusions:Epilepsy has a high prevalence in AD patients, and seizures may be a risk factor for progression from cognitive impairment to AD.Early identification of seizures and cognitive impairment may have a positive effect on delaying the progression of dementia.

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.