Objective To investigate the expression and significance of T-bet in experimental autoimmune uveoretinitis (EAU). Methods EAU was induced in 24 Lewis rats by immunization with retinal S-antigen (50 ?g) and complete Freund′s adjuvant, and another 4 rats were the healthy control. The rats with EAU were executed 7, 12, 15, 21 days after immunization, respectively. Immunohistochemical single and double staining were performed using monoclonal antibodies of T-bet or CD4 on the ocular wholemounts and the consecutive sections of the eye and spleen from both 24 immunized Lewis rats and 4 normal controls. The positive cells were counted under the optic microscope and the data were analyzed by SPSS 11.0 statistic software. Results A few T-bet positive cells were observed in the normal ocular tissues and spleen. The expressions of T-bet in the iris, retina, and spleen increased 7 days after immunization, reached the peak at the 15th day, and decreased at the 21st day, which were higher than those in the control. Double staining on the consecutive sections revealed that most of the T-bet positive cells were positive for CD4 monoclonal antibody. Conclusion T-bet may affect the occurrence of EAU by activating Th1 cells.

ObjectiveTo study the effects of perindopril on myocardial energy metabolism and ultrastructural changes in rats with chronic heart failure (CHF) induce by isoproterenol. MethodsTotally 55 male SD rats were randomly (random number) divided into two groups, namely control group (Group C) and CHF model group. The CHF rat models were made by subcutaneous injection of isopreteronol (ISO) in doses of 20 mg · kg-1 · d-1, 10mg · kg-1 · d-1 and 5 mg/kg/d for successive 3 days and then 3 mg· kg-1· d-1 for9 days. Four weeks later, the rats in CHF model group were randomly further divided into two subgroups, namely untreated subgroup (group M ) and perindopril treated subgroup (group P). After treatment for five weeks in average, echocardiography and myocardial pathology examination carried out to assess the cardiac function and structure changes of these rats. The levels of ATP, ADP, AMP, lactic acid (LA) and sarcoplasmic reticulum Ca2+ -ATPase (SERCA) activity in myocardium were determined by enzymatic reaction. ResultsCompared with rats in group M, the ejection fraction of left ventricle (EF) and fractional shortening of short axis of left ventricle (FS) of the rats in group P increased by 3.25％ and 7. 33％, respectively. Compared with rats in group C, the myocardial ATP, AMP, TAN (total adenosine) and LA significantly decreased in rats of group M. There were no significant differences in the levels of ADP, AMP, ATP/ADP and TAN between group C and group P (P ＞0. 05). Compared with rats in group M, the myocardial SERCA activity increased by 16. 41％ in rats of group P. The myocardial injury found under microscope and electronic microscope was ameliorated by treatment with peidolapril in rats of group P in comparison with rats of group M. ConclusionsPerindopril can improve myocardial energy metabolism,and lessen the pathological changes of ultrastructure, enhancing the cardiac function of rats with CHF induced by ISO.

Aim To investigate the influence and molecular mechanism of C-phycocyanin(CPC) from Spirulina platensis on apoptosis of HeLa cells in vitro.Methods Firstly,the effect of purified CPC on proliferation of HeLa cells in vitro was determined by MTT assay,and then electron microscope was exploited to observe the characteristic apoptotic features of cells treated with CPC.Subsequently,genomic DNA changes of HeLa cells were observed by agarose electrophoresis.Flow cytometric analysis revealed the influence of different concentrations of CPC on cell cycle of HeLa cells.The expressions of apoptosis related genes of CPC treated HeLa cells were determined by immunohistochemistry analysis.In addition,the activities of caspases and the release of cytochrome c from mitochondria into the cytosol were detected.Results Compared with control cells untreated with CPC,a significant decreased in the numbers of HeLa cells in survival treated with CPC and concentration dose effects existed.CPC could induce characteristic apoptotic features including cell shrinkage,membrane blebbing,microvilli loss,chromatin margination and condensation into dense granules or blocks.DNA of HeLa cells treated with CPC showed fragmentation pattern(DNA ladder of oligomers of 180~200 bp) typical for apoptotic cells.HeLa cells treated with different concentrations of CPC demonstrated an increasing percentage of cells in sub-G0/G1 phase.In addition,CPC could promote the expression of pro-apoptotic gene(Fas and ICAM-1);meanwhile,held back the anti-apoptotic gene(Bcl-2) expression,and then facilitated the transduction of tumoral apoptosis signals that resulted in the apoptosis of HeLa cells in vitro.In CPC treated HeLa cells,CPC treatment of HeLa cells also resulted in activation of caspases and release of cytochrome C from mitochondria into the cytosol.Conclusion C-phycocyanin from Spirulina platensis can induce the apoptosis of HeLa cells in vitro.By virtue of the promotion of the apoptosis signals transduction in HeLa,CPC realizes its antitumor activities.

BackgroundMonocyte chemotactic protein-1 (MCP-1)plays an important role in the tumor,inflammation,diabetic retinopathy and other neovascular disease,but the expression and the role of MCP-1 in the oxygen induced retinopathy(OIR) model have rarely been reported. Objective This study was to investigate the expression of MCP-1 in the retina development of newborn mouse and in mouse models with OIR.Methods C57BL/6J newborn mice were divided into two groups and 60 mice in each group.Mice in OIR group were exposed to 75％ oxygen for 5 days and then to room air.All mice in normal control group exposed to room air only.Ten mice in each group were randomly chosen and sacrificed at postnatal 5,7,12,14,17,21 days.The expression of MCP-1 in mouse retina was detected with the method of immunohistoehemistry and reverse transcription polymerase chain reaction(RT-PCR).Results MCP-1 positive cells were seen in normal mouse retina.Up-regulation of MCP-1 positive cells was detected both in 12 days in normal control group and in 14 days in OIR group.MCP-1 mRNA was detected in mouse retina at 5 days,and a transient up-regulation of MCP-1 mRNA was observed in 12 days in normal control group.MCP-1 mRNA in OIR group significantly increased in 14 days in comparison with the normal control group( P =0.028,P =0.001 ). Conclusions Expression of MCP-1 is detectable in whole retinal development procession of mice.A transient up-regulation of MCP-1 expression is detected in the critical period of retinal vascular development in mice models with OIR,which is closely related to the retinal vascular development and progression of retinal new vessels.

Background Various studies have suggested that inflammatory factors such as leucocytes and macrophages are involved in the occurrence and development of diabetic retinopathy (DR),and many cytokines promote the occurrence of DR.However,the relationship of aqueous and serum monocyte chemotactic protein-1 (MCP-1) and macrophage migration inhibitory factor (MIF) change with DR is unclear.Objective This study was to investigate the effects of MCP-1 and MIF in aqueous and serum during DR development.Methods Eighty patients with type 2 diabetes were enrolled from Beijing Shijitan Hospital.These patients received phacoemulsification or phacoemulsification and vitrectomy from September,2010 to June,2011.Twenty-six cataract patients in the same stage (without diabetes) who underwent phacoemulsification surgery served as controls.According to the clinical stage of the DR,the diabetic patients were classified as the non-DR group (NDR) (20 eyes),non-proliferative DR group (NPDR) (38 eyes) and proliferative DR group (PDR) (22 eyes).Aqueous humour and periphery blood samples were collected during the operation to detect MCP-1 and MIF using enzyme-linked immnunosorbent assay (ELISA).Written informed consent was obtained from each subject before any relevant medical examination.Results The average aqueous MCP-1 levels were(1660.78±562.98),(1463.26± 623.41),(686.76±186.16) and(494.35±148.59) ng/L in the PDR group,NPDR group,NDR group and control group,respectively,showing a significant difference among the 4 groups (F=37.968,P=0.000).No significant differences were found in the aqueous MCP-1 levels between the control group and NDR group (P=0.169),or between the NPDR group and PDR group (P=0.117).However,the aqueous MCP-1 levels were significantly elevated in the PDR group,NPDR group and NDR group compared with the control group (P=0.000).The average aqueous MIF levels were (6.85±1.99),(3.56±0.90),(1.10±0.48) and (0.86 ± 0.46) μg/L,respectively,with significant differences among them (F =144.502,P =0.000).Multiple comparisons between groups were found to be significantly different (P =0.000) according to the LSD-t test,except between the control group and NDR group (P =0.475).A significant positive correlation was seen between the aqueous MCP-1 level and MCP-1 level in all study participants (r =0.564,P =0.000).However,serum levels of MCP-1 and MIF were not statistically significantly different among the 4 groups (F =2.158,P＞0.05;F =0.813,P＞0.05).Conclusions The increase of the aqueous MIF and MCP-1 levels is associated with the progression of diabetic retinopathy.The results suggest that MIF and MCP-1 promote the occurrence of DR.

ObjectiveTo evaluate the therapeutic effect ofZishen decoction combined with standard treatment of western medicine for for stage IV diabetic nephropathy with Qi and Yin deficiency.MethodsA total of 112 patients with stage IV diabetic nephropathy and Qi and Yin deficiency were randomized to thestandard treatment and the combined treatment groups, 56 in each. The standard treatment group received conventional treatment, including blood glucose controlling, antihypertensive, blood lipid regulating and diet controlling. The combined treatment group receivedZishen decoction on the basis of conventional treatment. All the patients were treated for 3 months. The blood urea nitrogen (BUN), serum creatinine (SCr), total cholesterol (TG), triacylglycerol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and glycated hemoglobin (HbA1c) were measured by an automatic chemistry analyzer. The urinary albumin excretion rate (UAER) was measured by the enzyme-linked immunosorbent assay.ResultsCompared with the standard treatment group, the SCr (53.51 ± 18.12μmol/Lvs. 62.66 ± 21.14μmol/L;t=2.459,P<0.05), UAER(100.73±84.24μg/minvs. 156.24 ± 96.38μg/min;t=3.245,P<0.05), TG(1.73±0.22 mmol/Lvs. 2.06 ± 0.21 mmol/L;t=8.112,P<0.01), TC(4.56 ± 0.62 mmol/Lvs. 5.10 ±0.31 mmol/L;t=5.830, P<0.01), LDL-C (2.42 ± 1.05 mmol/Lvs. 3.31 ± 0.81 mmol/L;t=5.022,P<0.01) in the combined treatment group decreased significantly, and the HDL-C (1.67 ± 0.33 mmol/Lvs. 1.36 ± 0.41 mmol/L;t=4.460,P<0.01) increased significantly. ConclusionZishen decoction on the basis of conventional treatment can improve the SCr and UAER, and regulate the blood lipid in the patients with stage IV diabetic nephropathy and Qi-Yin deficiency.

Objective To evaluate the effect of the self-made Zishen decoction combined with conventional western medicine therapy and health education on the patients with early diabetic nephropathy. Methods The 112 patients with DN were randomly assigned to 2 groups (each group 56 patients) at a ratio of 1:1. The control group was treated with control of glucose, blood pressure, lipid, and diet therapy, and the treatment group was with self-made prescription of invigorating the kidney and health education based on the control group treatment. All patients were treated for 6 months. The SF-36 scale was used to assess the quality of life, and the clinical effect was determined based on the blood biochemical indexes. Results Total clinical effect of the treatment group was 85.7%(48/56), and the control group was 67.9%(38/56) (χ2=4.057, P=0.044). Compared with the control group after treatment for 6 months, the physical condition (72.17 ± 13.41 vs. 64.59 ± 11.83, t=3.172), social function (64.58 ± 14.54 vs. 58.94 ± 14.62, t=2.047), physical role function (55.82 ± 10.11 vs. 47.46 ± 10.18, t=4.360), emotional role function (60.43 ± 10.20 vs. 56.04 ± 11.44, t=2.143), energy (69.86 ± 11.43 vs. 62.47 ± 11.12, t=3.468), general health status (68.57 ± 11.25 vs. 62.45 ± 11.78, t=2.812) of the treatment group were significantly improved (P<0.01 or P<0.05). Conclusion The self-made Zishen decoction and health education combined with conventional western medicine can improve the clinical effect and quality of life of patients with DN.

OBJECTIVETo evaluate the accuracy of 3 impression materials in reproductions of simian dental arches using a 3-dimensional optical digitizer.

METHODSTwo simian dental arches were prepared as the master models. Impressions were made for stone casts using three impression materials, including alginate impression materials, C-silicone materials and Impregum-Penta polyether rubber. Five plaster replication models for each master model, as well as for each impression materials were made. The master models and the casts were digitized using a 3-dimensional optical scanner and digitizer. The images of each plaster cast and its original master model were superimposed to obtain the setting cross-section volume of the dental crown. The ratios of the plaster cast volume change and discrepancy distribution patterns were analyzed.

RESULTSCompared with the volume of the master models, the discrepancies of the plaster casts volume were -5.84%, -3.21%, and -0.63% for alginate impression materials, silicone materials and Impregum-Penta polyether rubber, respectively. The discrepancy between the master models and casts from alginate material was statistically significant (P<0.05), but not for silicone materials or Impregum-Penta polyether rubber. Maximal deviation of image fitting was located in the cervix and the gingival areas.

CONCLUSIONThe volumes of all the plaster casts from the 3 impression material are smaller than that of the master model. Impregum-Penta polyether rubber allows the most precise and silicone material the relatively accurate reproduction of the denture model, while alginate can not. The major error areas are in the dental cervix and gingival region.

Dental Casting Technique ; Dental Impression Materials ; chemistry ; Dental Impression Technique ; Dental Models ; Humans ; Imaging, Three-Dimensional ; methods

OBJECTIVETo study the effect of Buyang Huanwu decoction (BYHWD) inducing angiogenesis on the neuroblast migration from the subventricular zone and its mechanisms after focal cerebral ischemia.

METHODThe middle cerebral artery occlusion (MCAO) was performed to mice for 30 minutes to establish the model. The rats were divided into sham group, model group, BYHWD group and endostatin group. BYHWD (20 g x kg(-1), ig) and endostatin (10 μg, sc) were administered 24 h after ischemia once a day for consecutively 14 days. At 14 d after ischemia, the density of micro-vessel and the number of neuroblasts in the ischemia border zone were determined by immunofluorescence staining. The mRNA and protein expression of cell-derived factor-1 (SDF-1) and brain-derived neurotrophic (BDNF) were examined by real-time PCR and Western blot.

RESULTCompared with the model group, BYHWD significantly increased the density of micro-vessel and the number of DCX positive cells in the ischemia border zone (P < 0.01), and significantly increased the SDF-1 and BDNF mRNA and protein expression (P < 0.01). Compared with BYHWD group, endostatin significantly reduced the density of micro-vessel and the number of DCX positive cells in the ischemia border zone (P < 0.01), as well as the SDF-1, BDNF mRNA and protein expression (P < 0.01).

CONCLUSIONBYHWD could promote the neuroblast migration from the subventricular zone via inducing angiogenesis after cerebral ischemia, the mechanism may be correlated with up-regulating the expression of SDF-1 and BDNF.

Angiogenesis Inducing Agents ; pharmacology ; Animals ; Brain Ischemia ; pathology ; physiopathology ; Brain-Derived Neurotrophic Factor ; analysis ; genetics ; Cell Movement ; drug effects ; Cerebral Ventricles ; pathology ; Chemokine CXCL12 ; analysis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Neurons ; drug effects ; physiology

OBJECTIVETo explore the effect of ligustrazine on the migration of bone marrow mesenchymal stem cells (BMSCs) and protein expressions of matrix metalloproteinase-2 and-9 (MMP-2 and MMP-9) in vitro.

METHODSBMSCs were in vitro isolated and cultured using whole bone marrow adherent method, and phenotypes [surface positive antigens (CD29 and CD90) and negative antigens (CD34 and CD45)] identified using flow cytometry. BMSCs were divided into the blank control group, 25, 50, 100 µmol/L ligustrazine group, and the GM6001 group (100 µmol/L ligustrazine +MMPs inhibitor GM6001 ). The migration of BMSCs was tested by Transwell chamber test and wound healing assay after treated with ligustrazine for 24 h. The protein expressions of MMP-2 and MMP-9 were detected by Western blot.

RESULTSThe third passage BMSCs grew well in uniform morphology. The expression rate of CD29, CD90, CD34, and CD45 was 96.9%, 97.3%, 0.2%, and 3.0%, respectively. Compared with the blank control group, the number of migrated cells and relative distance of cell invasion increased, and the protein expressions of MMP-2 and MMP-9 were elevated in each ligustrazine group (P < 0.05, P < 0.01). Compared with 100 µmol/L ligustrazine group, the number of migrated cells and relative distance of cell invasion decreased in 25 and 50 µmol/L ligustrazine groups and the GM6001 group (P < 0.01). Protein expression of MMP-2 decreased in 25 and 50 µmol/L ligustrazine groups (P < 0.01).

CONCLUSIONLigustrazine could promote the migration of BMSCs in vitro, and its mechanism might be related to up-regulating expression levels of MMP-2 and MMP-9 protein.

Cell Movement ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Pyrazines ; pharmacology ; Up-Regulation