AIM: To develop a rat model of insulin resistance and observe the effect of puerarin injection on expression of GSK-3 in insulin resistance rats.METHEDS: 30 Wistar rats were divided into three groups randomly: control group,model group and puerarin group.The model group and puerarin group were given high fat diet(20% surcose,10% ard,(2.5)% chlesterol,1% cholic acid) for 4 weeks.Then the puerarin group was given abdominal injection puerarin 100(mg?kg~(-1)?d~(-1)) for six weeks.At the end of this experiment,we detect GSK-3 protein by western-blot analysis.Then computer image pattern analysis system was used to analyze the expression of GSK-3 in the hepatacyte.RESULTS: Tthe model group showed a hyperglycemia,hyperinsulinism,the ISI decreased and HOMA-IR increased.GSK-3 expression enhanced in the model group,but the expression of GSK-3 in puerarin group decreased compared to the model group.CONCLUSION: Puerarin injection can significantly decreased the expression of GSK-3 and improve insulin resistance.

Objective To study the relationship between GAT-1 and neuron injury in focal cerebral ischemia/infusion model.Methods Focal cerebral ischemia/reperfusion models were established in rats by thread embolic, then the brain tissues were taken at 3 h, 6 h,12 h, 24 h and 3 d after cerebral ischemia/reperfusion.The number of GAT-1 positive neurons were calculated and the average optical density were detected by immunohistochemical technique in different time points after cerebral ischemia/reperfusion, compared with the normal group and the sham operated group.Results The expression of GAT-1 was up-regulation obviously at 3 h after cerebral ischemia/reperfusion( P

Objective To investigate the relationship between COX-2 and Ki-67 proliferation index, bax, microvascular density(MVD)and CD44v6 respectively. Methods The expression of COX-2,Ki-67, bax, MVD (labelled by CD34) and CD44v6 were examined by immunohistochemistry PV method and tissue chip method in 101 cases of cervical carcinomas epithelium. Results The expression of COX-2 in cervical carcinomas has positive correlation with Ki-67, MVD and CD44v6, but has no correlation with bax. Conclusion The probable mechanisms are related with neovascularization, cell proliferation and cell adherence, but not related with the expression of bax.

Objectives To investigate the proliferation and apoptosis of smooth muscle cells(SMCs) after implantation of NiTi radioactive stents. Methods Radioactive and nonradioactive stents were implanted in abdominal aortas of 76 rabbits.The proliferation and apoptosis of SMCs were observed using immunohistochemistry and TUNEL.Results (1) Neointima areas on radioactive stents were less than those on nonradioacvtive stents at 1-month and 3-month respectively (P0 05).(2)The expressions of PCNA were lower in radioactive stent groups at different times compared with those of controls(P0.05).Conclusions (1) Radioactive stents can reduce the neointima areas on the implanted stents.(2)Radioactive stents can inhibit the proliferation of SMCs.(3)Radioactive stents promote the apoptosis of SMCs.

Objective To construct an immortalized rat astrocyte strain expressing enkephalin which can be regulated by doxycycline.Methods pRevTet-On and recombinant plasmid pRevTRE/hPPE were transfected into packaging cell PT67 respectively by standard lipofectamine methods.The supernatant containing RevTet-On virus was used to infect immortalized rat astrocyte strain(IAST)and RevTRE/hPPE virus was used to infect the IAST/Tet-On cell.Immortalized rat astrocyte strain that stably expressed Tet-regulated enkephalin was established. hPPE gene expression level of IAST/Tet-On/hPPE cell strain was detected by real time-PCR,immuno- cytochemistry and radio-immunoassay.Results Real time-PCR,immuno-cytochemistry and radio-immunoassay confirmed the level of enkephalin expression was regulated by doxycycline in a dose-dependent manner.Conclusion An immortalized rat astrocyte strain which secrets enkephalin as controlled by doxycycline has been constructed successfully.

Objective To investigate the effect of hyperuricemia(HUA) on inflammation and endothelin-1 (ET-1) production and treatment of Benzbnomanone in hypertensive patients.Methods 90 initial hypertensive patients were enrolled from the inpatient division and clinic of our hospital,60 patients of them were identified HUA,and 30 patients were normal in uric acid as control.All these hypertensive patients with HUA were treated with basic anti-hypertensive drugs,of them 30 patients were additionally treated with Benzbromarone table 50mg for 8 weeks.The levels of inflammation indices and ET-1 were compared between these hypertensive patients with HUA and hypertensive patients with normal serum uric acid,also hypertensive patients with HUA treated with or without Benzbromarone for 8 weeks.Results Compared with the hypertensive patients with normal serum uric acid,levels of ET-1,interleukin-1 (IL-1) and high-sensitive C reactive protein (hsCRP) were higher in the hypertensive patients with HUA.Also,the levels of these indices were positively correlated with the level of serum uric acid [(86.6 ± 4.8) pg/ml vs (82.4 ±6.9)pg/ml; (47.6 ±6.2)mg/L vs (19.1 ±4.1) mg/L; (3.4 ±0.8)mg/L vs (2.9 ± 1.1)mg/L,r =0.81,0.74,0.83,all P ＜ 0.05].Benzbromarone could effectively decrease the levels of ET-1,IL-1and hsCRP in the hypertensive patients with HUA [(49.8 ± 5.0) pg/ml vs (87.5 ± 5.9) pg/ml ; (17.6 ±8.8) mg/L vs (48.2 ± 7.0) mg/L; (1.7 ± 0.7) mg/L vs (3.5 ± 0.9) mg/L,all P ＜ 0.05].Conclusions HUA could increase the levels of inflammation and ET-1,while Benzbromarone effectivelv decreased these changes.Decreasing the level of serum uric acid would retard the process of atherosclerosis in the hypertensive patients with HUA.

Background Previous studies showed that the pathogenesis of uveitis is related to γδ T cells.However,it remains unclear that how these cells are involved in experimental autoimmune uveitis (EAU).Objective This study aimed to observe the dynamic changes of γδ T cells in EAU and explore the role of γδ T cells in the pathological process of EAU.Methods Forty-five C57BL/6(B6) mice were assigned to the normal control group (six mice) and EAU model group (thirty-nine mice).The mice were immunized subcutaneously at 6 spots on the footpads,tail base,and flank with emulsion containing human interphotoreceptor retinoid binding protein1-20 (IRBP1-20) emulsified in complete Freund's adjuvant.After immunization,the mice were examined for clinical signs of EAU by using a Genesis-D camera.The changes of histopathology were compared by hematoxylin and eosin staining.Mouse lymphocytes were isolated and purified from the spleens of IRBP1-20-immunized or normal B6 mice by using a γδ T-cell isolation kit.Flow cytometry was used to detect the changes of intracellular expression of interleukin-17A (IL-17A),and then transferred the activated γδ T cells into EAU models to analyze the changes of clinical signs and histopathology of EAU.Experimental study program as well as the use and feeding of the animals were authorized by the Animal Management and Use Committee of Shandong Traditional Chinese Medicine University.Results The inflammatory symptoms in mouse eyes appeared on day 12 after modeling.The initial changes were fundal blood vessel thickening and minimal inflammatory cell infiltration.Then,multifocal chorioretinal lesions,serious vasculitis and linear lesions were observed on days 16-20,along with abundant lymphocyte infiltration in the vitreous and retinal disorganization.The inflammation symptom scores and the pathological inflammation scores at different time points after modeling had statistically significant differences (F =51.399,P =0.000;F =47.342,P =0.000).The inflammatory symptoms in the eyes began to abate from day 28 onwards.The number of γδ T cells was obviously increased during the inflammation phase of EAU at day 16-20 after modeling,with the number of γδ T cells was (5.67 ±-0.49) ％ and (5.78 ±±0.55) ％,respectively,which was significantly higher than (1.53 ± 0.14) ％ before modeling,with significant differences between them (both at P＜0.05),meanwhile CD69 levels and the integrin lymphocyte function-associated antigen-1 (LFA-1) and secreted IL-17A were elavated.The secretion level of IL-17A was (13.40±0.50)％ and (17.80±2.37)％ on day 16 and day 20 after modeling,respectively,which was significantly higher than (1.53 ± 0.19) ％ before modeling,with significant differences between them (P =0.000,0.001).The activated γδ T cells were transferred into EAU model,the inflammation symptom scores were 1.00 (1.00,2.00) after activated γδ T cells were transferred into EAU model,which was significantly higher than 0.75 (0.05,1.00) of the untransferred group (Z =27.00,P =0.03),and the symptoms of EAU were aggravated.Conclusions The proportion of γδ T cells reaches peak in inflammation of EAU,and the cells are activated.The activated γδ T cells in the EAU model play a immune regulation role by secreting IL-17A.

Objective To detect the expression of β-catenin in the tissues of primary central nervous system lymphoma (PCNSL), and to discuss its function in PCNSL.Methods The paraffin embedded tissues from 10 patients diagnosed as PCNSL from October 2010 to April 2012 were collected as the experimental group.The paraffin embedded tissues from 10 patients with lymphadenitis were collected as the control group.Quantitative real-time PCR and immunohistochemical method were used to detect the expression of β-catenin in these tissues, and the relationships between β-catenin and clinical data were analyzed.Results Immunohistochemistry results showed that β-catenin protein was localized in the cytoplasm and (or) nucleus.Among 10 PCNSL patients, β-catenin protein was positive in 4 patients, while it was no positive in all of 10 lymphadenitis patients, with the significant differences between both groups (P ＜ 0.05).The β-catenin gene relative expression level was 4.70±0.57 and 1.00±0.27 in the experimental group and the control group, respectively.β-catenin expression was no correlation to age, PS score, cerebrospinal fluid protein level and serum lactate dehydrogenase level of patients with PCNSL.Conclusions Whether in mRNA level or in protein level, β-catenin expression is always high in PCNSL tissues, and its protein is expressed in the cytoplasm, however, this phenomenon was not observed in the tissue of lymphadenitis.

Objective To investigate the effect of the exogenous fragile hisdidine triad(FHIT) gene on the proliferation and the apoptosis of cutaneous carcinoma cell line A431,and to explore the mechanism of tumor suppression by the FHIT gene.Methods The plasmids pcDNA3-FHIT and pcDNA3-vector were transfected into the cutaneous carcinoma cell line A431 without FHIT gene expression,and then the transfected cells were screened by G418 and the expression of FHIT was determined by the immunocytochemical staining technique.The effect of FHIT on the growth characteristics of cutaneous carcinoma cell line A431 was observed by MTT,colony forming test and flow cytometry.Results Stable FHIT gene expressing A431 cells were produced,the proliferation activity and colony forming capability of A431FHIT were suppressed,whereas the apoptosis was increased.All these differences between A431-FHIT cells and the two control groups of cutaneous carcinoma cells had statistical significance.Conclusion Transfecting the exogenous FHIT gene into cutaneous carcinoma cells line A431can suppress the proliferation of tumor cells,and can also induce apoptosis and cell cycle arrest.

Objective To study the constituents in water-extracts from Flos Lonicerae. Methods Compounds were isolated and purified by using various column chromatography such as D101, Sephadex LH-20, and silica gel, etc. Their structures were identified on the basis of physical and spectral data. Results Seven secoiridoid glycosides were obtained and identified as vogeloside (Ⅰ), 7-epi-vogeloside (Ⅱ), secologanic acid (Ⅲ), sweroside (Ⅳ), secoxyloganin (Ⅴ), secologanoside (Ⅵ), (E)-aldosecologanin (Ⅶ). Conclusion Among them, compounds Ⅲ and Ⅵ are firstly obtained from the plants in Lonicera L. The structure of compound Ⅶ is rare in nature so far.