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 study the effect of cognitive therapy in combination with electromyographic feedback therapy on diabetes patients with depression after cerebral infarction. Methods: 60 patients were randomly divided into study group (n=30) and control group (n=30) , The former received cognitive therapy and electromyographic feedback trainings in addition to regular medication. All patients were evaluated with Hamilton Depression Rating Scales (HAMD),Neurological Function Deficit scale(NFD)?Barthel Index(BI) and FPG (free plasma glucose), 2HPG (plasma glucose after 2 hours of a meal). Results: The improvement of study group was greater than the control group(P

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 explore a three-dimensional examination method for retinal vessels. Design Experimental study. Participants Retinal vascular digest preparations of rat. Methods After deep anaesthesia, rats were sacrificed and perfused till eyeballs pale. Then 4% paraformaldehyde was perfused for vascular internal fixation. Eyeballs were fixed in 4% paraformaldehyde for 24hr, and the retinal vessels were isolated with collagenase digest technique. After immunofluorescence staining, the retinal vessels were observed with laser scanning confocal microscope. Main Outcome Measures The digest state of retinal vascular digest preparations and three-dimensional observation of the preparations. Results Ideal complete retinal vascular digest preparations were obtained after collagenase digestion. Three-dimensional characteristics of retinal vessels can be observed with laser scanning confocal microscope. Conclusions Modified retinal vascular digest preparations combined with laser scanning confocal microscope provided us a method for retinal vascular three-dimensional structure observation.(phthalmol CHN,2006,15:138-141)

Objective To investigate the expression level of serum Golgi protein-73 (GP73) in patients with primary hepatocellular carcinoma (HCC), or accompanied with Budd-Chiari syndrome (BCS), and to discuss its clinical value in making early diagnosis. Methods The peripheral serum samples were collected in 490 individuals, including patients with HCC (n = 65), cholangiocarcinoma (CC, n = 35), Budd-Chiari syndrome accompanied with HCC (n = 40), type B virus hepatitis (n = 60), cirrhosis after B hepatitis (n = 60), Budd-Chiari syndrome complicated by cirrhosis (n = 60), other malignant lesions (n =50), benign hepatic lesions (n = 60) and healthy volunteers (n = 60). The expression levels of serum GP73 and AFP were determined by using Elisa and electrochemiluminescence methods separately. Results The expression level of serum GP73 in the patients with HCC , CC or BCS plus HCC were significantly higher than that in the other individuals (P < 0.05). The area under the receiver operating characteristic curve (ROC), the sensitivity, the specificity and the diagnostic accuracy of GP73 and AFP for HCC were 0.868, 90.8%, 75.5%, 77.6% and 0.739, 60%, 83.2%, 80.2% respectively. The cut-off values of GP73 and AFP were 43.40 ng/ml and 20ng/ml respectively. The area under ROC and the sensitivity of GP73 for the diagnosis of HCC were strikingly higher than those of AFP (P < 0.05), while no significant differences in the specificity and diagnostic accuracy existed between GP73 and AFP (P > 0.05). The area under ROC, sensitivity, specificity and the accuracy of GP73 for the diagnosis of CC were 0.774 , 88.6%, 72.7% and 73.9%respectively, and the optimal cut-off value was 45.40 ng/ml. Conclusion In diagnosing HCC, the expression level of serum GP73 is superior to AFP. It is hopeful that serum GP73 may be proved to be an early serum tumor marker for HCC.

Objective A group of patients who had undergone total hip arthroplasty were followed up radiologically to explore the effects of abduction angle, the anteversion angle and the height of the femoral head center on dislocation, because these measurements could be used as a basis to evaluate current practice and to seek improvements. Methods A series of 326 hip joints (318 patients) were followed up routinely. The average follow up period was 2.7 years. 10 was found to have dislocation. The abduction angle, the anteversion angle and the height of the femoral head center were measured. The results were statistically analyzed, with a P value less than 0.05 indicating significant difference. Results The abduction angle of 55?or larger was found to be associated with greater risk of dislocation, compared with the abduction angle of less than 55?. Meanwhile, cups with the femoral head center superior placement greater than 30 mm resulted in more dislocation, compared with those less than 30 mm. As an independent variable, the anteversion angle had no significant association with dislocation. Conclusions Based on the results of the current study, the abduction angle and the height of the femoral head center should be considered as prerequisites for maintaining hip stability and decreasing the risk of dislocation. We believe that hips with the abduction angle of less than 55?and the height of the femoral head center less than 30 mm can decrease the risk of dislocation.

Objective To investigate the clinical, hemorheology, transcranial Doppler (TCD) and auditory brainstem evoked response (ABR) features in senile sudden deafness.Methods The clinical manifestations, hemorheology, TCD, and ABR data in 82 patients with senile sudden deafness were analyzed retrospectively.Results (1) The primary symptoms mainly included tinnitus (65.9%) and vertigo (56.1%), which mostly occurred at daytime (53.7%).(2) Most of the patients had risk factors of arteriosclerosis such as hypertension (75.6%), diabetes (59.8%) and so on. These patients had more serious deafness.(3) There was no obvious relationship between high blood viscosity (61% of the patients) and the degree of deafness. (4) 85.4% of senile sudden deafness patients indicated vertebral-basilar artery insufficiency in TCD. There was a positive correlation between blood supply and degree of deafness. (5) The abnormal rate of ABR was 100%. WaveⅠdisappeared in most patients, which showed serious damage of auditory nerve. Disappearing of waveⅠ, Ⅲ, Ⅴ and Ⅰ～Ⅲ, Ⅲ～Ⅴ, Ⅰ～Ⅴ delitescence period delaying might be complicated with the damage of the structure of the brainstem and cochlea.Conclusions The morbidity and degree of senile sudden deafness are associated with insufficiency of blood supply of internal ears. TCD and ABR are sensitive and reliable methods in diagnosis, progression and prognosis of senile sudden deafness.

0.05).Conclusions The level of plasma Hcy in BD patients are higher than that of in the controls.Hyperhomocysteine may take part in the pathogenesis of BD, but the MTHFR gene C677T genotypes are not associated with BD.

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.

Based on untermolecular splut G-quardruplex-hemun DNAzymes, a buosensor for detectuon of sulver uons and cysteune was developed wuth magnetuc nanopartucles ( MNPs ) as carruer to ummobuluze the DNA probes. Sunce Ag+ chelates guanune bases at the bundung sutes whuch are unvolved un G-quadruplex formatuon, the presence of Ag+ may unhubut G-quartets connected by Hoogsteen-type base paurung and dusrupt G-quadruplexes structures, whuch decreases the peroxudase actuvuty of G-quadruplex-hemun DNAzymes that effucuently catalyze H2 O2-meduated reactuons, such as the oxudatuon of ABTS ( 2, 2'-azunobus ( 3-ethylbenzothuozolune)-6-sulfonuc acud) by H2 O2 . Moreover, un the presence of L-cysteune, ut was used as a competutor by the strongly Ag-S to release Ag+ from G-ruch olugonucleotudes, promotung the reformatuon of G-quadruplexes and uncreasung the peroxudase actuvuty, whuch catalyzes the ABTS-H2 O2 reactuon system. In thus experument, the effucuent separatuon from real sample was achueved usung magnetuc nanopartucles as a solud phase carruer to effectuvely uncrease the detectuon sensutuvuty and decrease the background sugnal. Under the optumum condutuons, a hugh lunear relatuonshup between the UV absorbance and the Ag+ concentratuon was establushed un the range of 0. 5-100 nmol/L wuth a detectuon lumut of 0. 2 nmol/L. The calubratuon curve of cysteune was udentufued un the range from 0. 1 to 80 nmol/L and the detectuon lumut was as low as 0. 04 nmol/L.