Diagnostic Techniques and Procedures ; instrumentation ; Humans ; Pathology, Clinical ; instrumentation ; methods ; trends ; Referral and Consultation ; Reproducibility of Results ; Sensitivity and Specificity ; Telepathology ; instrumentation ; methods ; trends

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 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 evaluate the value of different methods of hormone measurement and image study before surgery.Methods We retro spectively reviewed 99 patients with insulinoma confirmed by pathological report after surgery in the PUMC Hospital from December 1989 to August 2005.Results The main clinical manifestations were hypoglycemia coma,weight gain,perspiration,disorientation,hypomnesis,and dizziness.The sensitivity of diagnosis of insulinoma with fasting blood-glucose less than 2.78 mmol/L was 66.6%.The ratio of serum insulin to blood-glucose greater than 0.3 when hypoglycemia attacks was 90% sensitive.The sensitivity of perfusion CT was highest which was 100%.Conclusion Typical “Whipple's triad” and the ratio of serum insulin to blood-glucose greater than 0.3 when hypoglycemia attacks are best for the qualitative diagnosis of insulinoma.Perfusion CT has the best sensitivity and specificity among image studies currently used.

Objective To investigate the expression of IL-10 and IL-6 in tissue and plasma of subtypes lymphoma and its significance. Methods Immunohistochemical technique and enzyme-linked immunosorbent assay (ELISA) were used to assess the expression of IL-10 and IL-6 in 97 cases paraffin tissue and plasma samples from different subtypes of lymphoma.Results Expression of IL-10 and IL-6 was positive in all lymphoma tissue samples, but there is no significant difference among different subtypes lymphoma (x2 =0.815, x2 =0.542, P ＞ 0.05). Tumor samples were found positive IL-10 and IL-6 expression in macrophages and vascular endothelial cells by immunohistochemistry respectively. Expression of IL-10 and IL-6 in plasma of lymphoma were higher than that in control respectively [(232.57±191.59) pg/ml vs (59.12±68.35) pg/ml,(80.70±89.68) pg/ml vs (45.68±33.66) pg/ml],with significant difference (t =6.968,t =2.896,P ＜0.05).IL-10 and IL-6 plasma levels increased with enhanced expression in lymphoma (x2 =0.815,x2 =0.542,P ＜ 0.05).Which was found to be correlated with each other (rs =0.394,P ＜ 0.05). Conclusion The expression of IL-10 and IL-6 had been found at various degrees in tissues and plasma of lymphoma.IL-10 and IL-6 in plasma of lymphoma were higher than that in controls. These results showed that IL-10 and IL-6 may cooperate in vivo with lymphoma cell proliferation and infiltrate. IL-6 and IL-10 maybe served as useful indicators for diagnosis of lymphoma.

Objective To investigate if insulin can affect the expression of vascular endothelial growth factor (VEGF) in the retina of streptozotocin-induced diabetic rats. Methods A total of 60 male Sprague-Dawley rats were randomly divided into sodium citrate buffer control group (CIT-CON, n= 30) and STZ-induced diabetic group (STZ-DM, n=30). At the 16th week, 24 rats from CIT-CON group at random were randomly divided to group A (sodium citrate buffer control group, n = 12) and group B (sodium citrate buffer plus insulin group, n= 12). The remaining 6 rats from as CIT-CON group served as negative control. At the same time, 24 rats from STZ-DM group at random were randomly divided to group C (STZ-induced diabetic group, n= 12) and group D (STZ-induced diabetic plus insulin group, n= 12). The remaining 6 rats from STZ-DM group also served as negative control. 4 IU of insulin was injected subcutaneously to rats of group B and D. Immunohistochemistry, Western blot and Real-time polymerase chain reaction (RT-PCR) were used to measure the expression level of VEGF protein and mRNA respectively. RESULTS Insulin significantly increased the VEGF mRNA (7.71 ± 0.25 vs 5.36 ±0. 37, t test P< 0. 05) and protein expression (0. 4925 ± 0. 0122 vs 0. 4272 ± 0. 0110, t test P< 0. 05) in the retina of CIT-CON rats.However, in retina of STZ-DM rats, insulin had no effect on VEGF mRNA (8. 92±0. 27 vs 9. 05±0. 28, t test, P>0. 05) and protein expression (0. 5152±0. 0109 vs 0. 5099±0. 0100, t test P>0.05). Conclusions Insulin had no effect on VEGF expression in the retina of STZ-DM rats.

ObjectiveTo investigate the clinicopathologic characteristics of different nation nonHodgkin lymphoma (NHL)in Urumqi of Xinjiang Uygur Autonomous Region.MethodsFour hundred and sixty-six cases of NHL were collected.Their sections including HE and immunohistochemical staining were examined again for diagnosis and classification.ResultsAmong 466 cases NHL,B cell neoplasm was 369cases (79.2 ％) and T cell lymphoma was 97 cases (20.8 ％).193 cases (41.4 ％) occured in the lymph node and 273 cases (58.6 ％) extranodal lymphoma.The most common subtypes were diffuse large B-cell lymphoma,small lymphocytic lymphoma/chronic lymphocytic leukaemia,extranodal marginal zone lymphoma of mucosaassociated lymphoid tissue,NK/T cell lymphoma,peripheral T-cell lymphoma,follicular lymphoma.Tlymphoblastic lymphoma (T-LBL) and anaplastic large cell lymphoma (ALCL) in the Uygur were higher than the Han [7.5 ％ (9/120) vs 1.3 ％ (4/308),x2 =11.276,P=0.001; 4.2 ％ (5/120) vs 2.3 ％ (7/308),x2 =1.137,P =0.286],and T/NK cell lymphoma in the Han was higher than the Uygur [7.1 (22/308) vs 3.3 ％ (4/120),x2 =2.196,P =0.138].There was no statistically significance between the Uygur and Han in different subtypes of B-NHL (P ＞0.05).ConclusionIn Urumqi aera,the incidence is higher in extranodal lymphoma than that in nodal.B-NHL is more in Urumqi area than that in other area in our country.There is no difference obviously for mobility B-NHL in the Uygur and Han,but T-LBL and ALCL in the Uygur are higher than those in the Han,and T/NK cell lymphoma in the Han is higher than that in the Uygur.It is suggested to confirm further whether the different associated with different nation and region.

Objective To explore the influence of coal-arsenic exposure on human T cells proliferation and its mechanism.Methods Blood samples colleoted from individuals which lived in arsenism area of coal-burning type and non-arsenism area in Guizhou Province were divided into exposed group(17),mild(35),moderate(38) and severe arsenism group(19)and control group(35)according to Diagnosis Smndard for Endemic Arsenism (WS/T 211-2001).T cell stimulation index wag determined by methyl thiazolyl tetrazolium(MTT)colorimetric method.The intracellular Ca2+ exponential(IECa2+)in peripheral blood mononuclear cell(PBMC)was analyzed by Fho-3/AM dye and flow cytometry.DNA binding activity of actively T cells nuclear factor(NF-AT)in PBMC was evaluated by electrophoretie mobility shift assay(EMSA).Results Concanavalin A(ConA)stimulation decreased the T cells stimulation indexes in exposed group,mild,moderate and severe arsenism groups(1.315±0.962, 1.611±1.224,1.114±0.545,1.289±0.875)compared with control group(2.322±1.241),all the differences being statistically significant(P＜0.01).After stimulated by anti-CD3 monoclonal antibody(McAb),the T cells stimulation index in exposed group,mild,moderate and severe arsenism group(0.997±0.177,1.103±0.291,1.007±0.221, 0.957±0.205) were lower than that of control group(1.842±0.429,P ＜ 0.01 ). IECa2+ of PBMC after treated by anti-CD3 McAb in mild,moderate and severe arsenism group( 110.130±49.637,92.429±31.191,77.640± 35.372) were lower compared with control group(145.986±59.450,P ＜0.01 ). Moreover,IECa2+ in moderat and severe arsenism group were lower than exposed group(121.337±46.410,P ＜ 0.05). DNA binding activity of PBMC NF-AT in mild,moderate and severe arsenism group(1.354±0.446,1.290±0.291,1.159±0.411 ) were lowered than that of control group(1.722±0.291,P ＜ 0.01) and exposed group(1.611±0.294,P ＜ 0.05). Conclusions The coal-arsenic exposure can reduce the human T cells stimulation indexes,IECa2+ in PBMC and the DNA binding activity of NF-AT. It suggest that arsenic may suppress the proliferation ability of human T cells,which may be partly related to the influence of arsenic on T cell receptor(TCR)/CD3 signal transduetion pathway.

Objective To investigate the activation of T lymphocytes in human peripheral blood and the signaling molecules in protein kinase C/nuclear factor KB(PKC/NF-κB) pathway expressivity or activity changes in human peripheral blood mononuclear cells(PBMCs) exposed to coal-arsenic,to explore the role of PKC/NF-κB signal pathway in activation of T cells in human exposed to coal-arsenic. Methods Blood samples were collected from individuals who lived in arsenism area of coal-burning in Guizhou province, and were divided into asymptomatically exposed group (12),mild arsenism group (33),moderate arsenism group (34) and severe arscnism group (15) according to Diagnosis Standard for Endemic Arsenism (WS/T 211-2001). The individuals who lived in non-arsenism area were control group(27). The ratio of activated T ceils was analyzed by flow cytometry. DNA binding activity of NF-κB in PBMCs was evaluated by electrophoretic mobility shift assay(EMSA). The expression of PKCθ and phospho-PKCθ(pPKCθ) in PBMCs were detected with western blotting analysis. Results The ratio of activating T cells in asymptomatically exposed group[(21.76±15.31)%],mild arsenism group[(18.41±11.36)%],moderate arsenism group[(17.78±11.93)%]and severe arsenism group[(18.79±13.38)%]were all higher than that of control group[(3.19±2.12)%],the difference among all groups being statistically significant(F = 7.893,P < 0.05). DNA binding activity of PBMCs NF-κB in asymptomatically exposed group,mild arsenism group,moderate arsenism group and severe arsenism group(1.49±0.24,1.58±0.30,1.57±0.34,1.51±0.16) were higher than that of the control group(1.30±0.17),the difference being statistically sign/ficant(P < 0.05 or < 0.01). The expression of PBMCs pPKCθ in mild arsenism group,moderate arsenism group and severe arsenism group(0.64± 0.14,0.64±0.27,0.62±0.12) were all lower than that of the control group(0.93±0.20),the difference being statistically significant(P < 0.05). There were significant negative correlations between the expression of pPKCθ and the activity of NF-κB(r =-0.565,P < 0.01). There were significant positive correlations between the activity of NF-κB and the ratio of activating T cells(r = 0.546,P < 0.01). Conclusion Coal-arsenic enhances the DNA binding activity of NF-κB,reduces the expression of PBMCs pPKCθ in human PBMCs and up-regulates the activity of T cells. It suggests that the PKC/NF-κB signal might be one of transduction pathway via activating of T cells by coal-arsenic.

Animals ; Gene Expression Profiling ; Mitochondria, Heart ; genetics ; metabolism ; Myocardial Reperfusion Injury ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley