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 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 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.

Twelve compounds were isolated from the aerial parts of Achillea alpina by column chromatographies on silica gel, Sephadex LH-20, and semi-preparative HPLC. The structures were elucidated on the basis of spectral analysis. The compounds were identified as pellitorine(1), 8,9-dehydropellitorine(2), (E,E)-2,4-undecadien-8, 10-diynoic acid isobutylamide(3), (E,E)-2,4-tetradecadien-8,10-diynoic acid isobutylamide(4),sintenin(5), 4',5,7,8-tetramethoxyflavone(6), chrysoplenetin(7), formononetin(8), aurantiamide(9), asperglaucide(10), artemetin(11), and eupatorin(12). compounds 1-5 were isolated from this plant for the first time, and compounds 6-10 were isolated from the genus Achillea for the first time.
Achillea ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Enhancing medical students' humane quality education is an urgent requirement for modem medical mode transformation for medical education.The pharmacology teachers of Chongqing Medical University follow the modem education concepts and fully search the human spirit materials hidden in pharmacology,then actively explore how to integrate the humanity spirit education into the pharmacology teaching to achieve the changes of from exam-oriented education to quality education.

AIMTo investigate the protective effects and mechanism of IPC on myocardial ischemia/reperfusion injury.

METHODSEffects of IPC on arrhythmia and coronary blood flow and the release of AST, CPK, LDH, SOD and LFO at different time after ischemia/reperfusion injury in rat Langendorff hearts were studied.

RESULTSIPC decreased the release of AST, CPK and LDH and increased myocardial SOD activity and decreased LPO level. IPC also inhibited ischemia/reperfusion arrhythmias and increased coronary blood flow.

CONCLUSIONThe results showed that IPC had well protective effects on myocardial ischemia/reperfusion injury.

Animals ; Female ; Heart ; physiopathology ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Myocardial Reperfusion Injury ; prevention & control ; Rats ; Rats, Wistar

OBJECTIVETo study whether Langchuangping granule (LG) could exert its renal protection by down-regulating monocyte chemoattractant protein-1 (MCP-1) via suppressing nuclear factor kappa B (NF-kappaB) signaling pathway in BXSB lupus nephritis (LN) mice. Methods Eighteen male 11-week-old BXSB LN mice were randomly divided into three groups, i.e., the model group, the hormone group, and the Chinese medicine group, 6 in each. They were administered by gastrogavage with normal saline, methylprednisolone, and LG, respectively. Another six C57BL/6 male mice of the same age was taken as the normal control group, which was administered with normal saline by gastrogavage. All mice were treated once daily, for 4 successive weeks. The 24-h urine protein was determined. The mRNA and protein expressions of MCP-1 in the renal tissue were detected using RT-PCR and Western blot. The expression of NF-kappaB p65 in the renal tissue was detected using immunohistochemical assay. Activity index (AI) of the renal tissue was counted using PAS stain. The content of ds-DNA antibody was detected using ELISA. The correlations of the aforesaid indices were analyzed.

RESULTSThe 24-h urine protein level, serum ds-DNA antibody content, protein and mRNA expressions of MCP-1, NF-kappaB p65 expression level, and AI count were obviously higher in the model group than in the normal control group (P < 0.01). The aforesaid indices all obviously decreased after medication in the Chinese medicine group and the hormone group (P < 0.05). MCP-1 protein expression level was positively correlated with MCP-1 mRNA, NF-kappaB p65, AI, 24-h urine protein, and ds-DNA antibody of all LN mice (r= 0.984, 0.936, 0.887, 0.698, 0.679, all P < 0.01).

CONCLUSIONSLG possibly played renal protection by down-regulating NF-kappaB-mediated MCP-1 expression levels. MCP-1 played important roles in the occurrence and development of LN, being one of ideal targets for LN treatment.

Animals ; Chemokine CCL2 ; metabolism ; Disease Models, Animal ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; drug effects ; metabolism ; Lupus Nephritis ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; metabolism ; Signal Transduction ; drug effects

Actins ; immunology ; metabolism ; Antibodies, Monoclonal ; metabolism ; Child ; Diagnosis, Differential ; Fibroma ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Gastrointestinal Neoplasms ; metabolism ; pathology ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; Humans ; Leiomyoma ; metabolism ; pathology ; Male ; Pyloric Antrum ; pathology ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Objective To explore whether the degree of 99Tcm-methoxyisobutylisonitrile (MIBI) uptake in non-small cell lung cancer (NSCLC) could be correlated with the treatment response to three-dimensional conformal radiotherapy.Methods A total of 102 patients with NSCLC were studied with 99Tcm-MIBI SPECT before radiotherapy.The patients were classified by a follow-up CT as responders (complete or partial remission) and non-responders (stable or progressive disease).After intravenous administration of 740 MBq 99Tcm-MIBI, SPECT imaging at 10-30 min (early) and 2-3 h (delayed) were performed.Region of interest (ROI) was placed over the tumors and contralateral normal lung tissue.The uptake ratio of tumor to contralateral normal lung (T/N) was obtained from both early (ER) and delayed (DR) SPECT images.The retention index (RI) was measured as:RI = (DR - ER)/ER×100%.Statistical analysis was performed by two independent-sample t-test and Mann-Whitney U test using software SPSS 13.0.Results 99Tcm-MIBI uptake was significantly higher in responders than in non-responders:2.36 ±0.17 vs 1.82 ±0.14 (ER) and 2.48 ± 0.20 vs 1.94 ± 0.16 (DR), respectively (t = - 13.1,- 12.7, both P＜ 0.05).The median RI in the responders group was also significantly higher than that in the non-responders group (6.60% vs 5.13%, z = - 6.83, P ＜ 0.05).Conclusion ER, DR and RI of 99Tcm-MIBI SPECT might be useful to estimate the treatment response to three-dimensional conformal radiotherapy in patients with NSCLC.

OBJECTIVETo study the correlation between loss of heterozygosity (LOH) on chromosome 10q and pathologic features, pathogenesis, prognosis of astrocytic tumors.

METHODSLOH on 10q was studied by interphase fluorescence in-situ hybridization (FISH) in 85 cases of astrocytic tumor, including 35 cases of WHO grade II tumors and 50 cases of WHO grade IV tumors.

RESULTSLOH on 10q was detected in 6 cases (17.1%) of diffuse astrocytoma (WHO grade II) and 34 cases (68.0%) of glioblastoma (WHO grade IV). 10q polysomy was detected in 7 cases (20.0%) of diffuse astrocytoma and 11 cases (22.0%) of glioblastoma. The rates of LOH on 10q in young age group and elderly group were 36.4% (12/33) and 82.4% (28/34), respectively. The difference was of statistical significance (P < 0.05). The rates of LOH on 10q in the diffuse astrocytoma and glioblastoma were 21.4% (6/28) and 87.2% (34/39), respectively. The difference was also statistically significant (P < 0.05). Univariate survival analysis showed that patient age, pathologic grade and 10q on LOH correlated with duration of survival (P < 0.05).

CONCLUSIONSThere are correlation between 10q LOH, patient age and pathologic grade of astrocytic tumors. LOH on 10q is also related to the pathogenesis of astrocytic tumors and is helpful in predicting prognosis.

Adolescent ; Adult ; Age Factors ; Astrocytoma ; genetics ; pathology ; surgery ; Brain Neoplasms ; genetics ; pathology ; surgery ; Child ; Chromosomes, Human, Pair 10 ; genetics ; Female ; Follow-Up Studies ; Glioblastoma ; genetics ; pathology ; surgery ; Humans ; Loss of Heterozygosity ; Male ; Middle Aged ; Neoplasm Grading ; Survival Rate ; Young Adult