Humans ; Root Resorption

Objective To observe the effects of grape seed proanthocyanidin extract(GSPE) plus atorvastatin treatment on matrix metalloproteinase-9 ( MMP-9) in experimental atherosclerosis rabbits and to explore its mechanism. Methods Thirty male New Zealand rabbits were randomized into five groups. The normal control group were fed with standard diet for 24 weeks. And the other groups were fed with standard diet containing 1 % cholesterol for 12 weeks. In the sequential 12 weeks, the model control group was fed with standard diet. The GSPE group was fed with standard diet containing 1% grape seed proanthocyanidin(GSP). The atorvastatin group was fed with standard diet containing atorvastatin(2. 5 mg · kg~(-1) · d~(-1)). The GSPE plus atorvastatin group was fed with standard diet containing 1% GSP and atorvastatin (2. 5 mg~(-1) · kg~(-1) · d~(-1)). Blood samples were drawn from ear middle arteries of rabbits just before the experiment and at the 12th and the 24th weekend of the experiment. All the rabbits were fasted for at least eight hours before the blood was drawn. The blood samples were analyzed for the matrix metalloproteinase-9 (MMP-9). All the rabbits were sacrificed at the 24th weekend, and the expression of MMP-9 was observed in the thoracic aortic tissue using immunohistochemistry technique. Results The serum level and aorta expression of MMP-9 were increased in model group compared to control group (all P< 0. 05). The severity of atherosclerosis was less in three drug groups than that in model control group. The GSPE,atorvastatin and GSPE plus atorvastatin groups versus model group showed less atherosclerotic lession, the decreased expression of aorta MMP-9 and the decreased serum level of MMP-9 C(1. 06±0. 21), ( 1. 07 ±0.20), (0.81 + 0.16) vs. (1. 32±0. 24)ng/ml,all P<0. 05]. The effect in the GSPE and atorvastatin group was most obvious. Conclusions GSPE plus atorvastatin group has the most efficacy of anti-atherogenesis, which is associated with its reducing the expression of matrix metalloproteinase.

Objective: To evaluate the quality consistency of the original product ( DIOVAN ) and generic valsartan capsules ( VSC) . Methods:The dissolution of the two capsules in four media was monitored by the UV method described in Chinese Pharmaco-poeia ( ChP 2010 edition) . The f2 similarity factor approach was used to evaluate the dissolution similarity between DIOVAN and VSC and the production quality of the enterprises. Meanwhile, the homogeneity in the same batch was studied by dissolution precision and the reproducibility in the different batches were also evaluated by a similar equivalent limit method to show the production quality of the manufacturers. Results:The dissolution of VSC and Diovan in pH 6. 8 phosphate buffer medium was both above 85% in 15 min. The f2 similarity factors in the other three media ( water, pH 4. 5 acetate buffer and 0. 1 mol·L-1 HCl) were all above 50. The relative standard deviation ( RSD) of precision in the same batch was less than 10%. Among the different batches, dissolution limit value ( Q) was within the range of the upper and lower limit value of probability levels (δ) . Conclusion:The f2 similarity factor results indicate the in vitro dissolution of the reference drug ( DIOVAN) and generic drug ( VSC) is consistent in the four media. The production quali-ty of generic manufacturer is also good with promising homogeneity and reproducibility evaluation results.

Ranibizumab is a recombinant rat monoclonal antibody fragment of the second generation humanized VEGF. Ranibi-zumab can bind all of active VEGF-A to inhibit the bond between VEGF-A and VEGFR1 or VEGFR2. As the results, the vascular en-dothelial proliferation and vascular permeability can be reduced, and the new vessels angiogenesis can be prevented. The article sys-tematically searched and summarized the relative literatures to offer high-quality evidences for the recent clinical application of ranibi-zumab in ophthalmology.

Objective To observe the subfoveal choroidal thickness (SFCT) and choriocapillary blood flow area (CBFA) in the patients with idiopathic macular hole (IMH).Methods This is a prospective clinical study.Thirty-two patients with unilateral IMH (4 in stage 2,17 in stage 3,11 in stage 4) and 32 age-and sexmatched normal controls were enrolled in this study.All eyes were divided into three groups,including group A (32 affected eyes),group B (32 fellow eyes) and group C (32 normal eyes of controls).There was no significant difference in age (t=0.865) and gender (x2=0.000) in IMH patients versus normal control subjects (P＞0.05).There was no significant difference in refraction (F=0.957) and ocular axial length (F=0.562) between group A,B and C.The SFCT was detected by enhanced depth imaging of spectral-domain optical coherence tomography (OCT).The CBFA was detected by OCT angiography.The differences of SFCT and CBFA in three groups were analyzed by Kruskal-Wallis and non-parametric test.Results The mean SFCT was (182.53 ±64.52) μtm in group A,(199.21 t73.07) μtm in group B and (254.21 ±56.85) μtm in group C respectively.The SFCT was thinner in group A and B than that in group C (Z=-4.362,-3.190;P＜0.05),but was the same in group A and B (Z=-1.171,P＞0.05).The mean CBFA was (5.09±0.31) mm2 in group A,(5.41 ±0.20) mm2 in group B and (5.39±0.15) mm2 in group C respectively.The CBFA was reduced in group A than that in group B and C (Z=-4.467,-4.048;P＜0.05),but was samc in group B and C (Z=0.420,P＞0.05).Conclusion SFCT and CBFA are both reduced in IMH eyes.

Objective To study the effect of astragaloside on TGF-β1,SMAD2/3,and α-SMA expression in the kidney tissue of diabetic KKAy mice,and evaluate its potential role in renal interstitial fibrosis.Methods 20 type 2 diabetic KKAy mice were randomly divided into model group and astragaloside group,while 10 male C57BL/6J mice were selected as the control.Astragaloside at 40 mg · kg-1 · d-1 was given when the KKAy mice fed with high-fat diet to 14 weeks old.The mice in the control and model group received normal saline at 40 mg · kg-1 · d-1.Blood glucose meter was used to detect the blood glucose value of each mice at 16th,20th and 24th week.The mice were killed at 24 weeks old and the kidney tissue samples were collected.Pathology morphological changes were observed.Results (1) blood glucose value:cmpared with the control group,the blood glucose value of KKAy mice at 14 week increased significantly,and that of model group also increased significantly at 16th,20th and 24th week (P＜0.05);the blood glucose value of astragaloside group decreased compared with control group (P＜0.05).(2) Morphology of kidney:in the control group,the glomerular and tubular had clear structure,there was no renal interstitial fibrosis;in the model group,the renal glomerular mesangial matrix had broaden,mesangial cell had increased,renal tubular epithelial cell cytoplasm showed vacuole degeneration,renal interstitial inflammatory cell had increaised.In astragaloside group,there were few renal tubular epithelial cell cytoplasm,and there was no obvious fibrosis.(3)TGF-β1,SMAD2/3,and α-SMA expression levels of the kidney issuse:compared with control group,mice in model group up-regulated TGF-β1,SMAD2/3 and α-SMA expression (P＜ 0.05).TGF-β1,SMAD2/3,and α-SMA expression levels in astragaloside group were significantly lower than those in the model group (P＜0.05).There was few phosphorylated SMAD2/3 expression in renal tubular and glomerular nuclei,while that of model group increased (P＜0.01),and compared with model group,that of the astragaloside group decreased (P＜0.05).Conclusion Astragaloside can delay the renal fibrosis process in diabetic mice by influencing the TGF-β/SMADS signaling pathway and down-regulating TGF-β1 and α-SMA expression,thus to relieve renal fibrosis in diabetic mice.

Objective To study the expression of signaling lymphocytic activation molecule (SLAM)CD150 in peripheral blood mononuclear cells(PBMCs)isolated from adult non-responders to recombined yeast gene hepatitis B vaccine.Methods A total of 202 cases were recruited.All these subjects had been immunized with recombined yeast gene hepatitis B vaccine for more than one standard scheme in two years(from Sep 2007 to Dec 2009)and remained negative for hepatitis B markers(HBsAg,anti-HBs,HBeAg,anti-HBe and anti-HBc).After recruitment,all 202 subjects received another standard scheme(0,1 and 6 month)revaccination.The blood samples were collected 7 months later after the first injection of revaccination to detect anti-HBs titer.The PBMCs were isolated from 18 adult non-responders(anti-HBs titer<10 mIU/mL)and 18 adult responders(antiHBs titer≥100 mIU/mL).CD150 expression on cell surface was analyzed by flow cytometry.SAS package was used for t test and spearman rank correlation analysis.Results After rHBsAg stimulation,the percentage of PBMCs expressed CD150 was significantly higher in non-responders (39.20%±10.66%)than responders(23.73%±12.41%)(t=2.1947,P<0.05).The same trend was also observed in rHBsAg stimulated C133+CD4+T cells,but the difference was not statistically significant(49.64%±11.94%vs 37.73%±11.02%)(t=1. 7175,P>0.05).After phytohaemagglutinin (PHA)stimulation,the percentage of CD150-positive PBMCs was also significantly higher in non-responders (39.21%±7.37%)than responders(23.18%±12.68%)(t=2.2835,P<0.05).CD150 expressions in both PBMCs and CD3+CD4+T cells were negatively correlated with anti-HBs titer after rHBsAg stimulation (r=-0.726,P<0.05).Conclusion Activation of CD150 may contribute to the non-response to hepatitis B vaccine.

Angiotensin II ; pharmacology ; Calcium ; metabolism ; Cell Hypoxia ; drug effects ; physiology ; Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; physiology ; Humans ; Membrane Potential, Mitochondrial ; drug effects ; physiology ; Oxygen ; metabolism

Objective To evaluate the effect of dexmedetomidine and small dose of ketamine on the expression of P2X4 receptor (P2X4 R) mRNA and P2X7 receptor (P2X7R) mRNA in the dorsal root ganglion of rats with neuropathic pain.Methods Ninety male Sprague-Dawley rats,aged 6-9 weeks,weighing 180-220 g,were randomly divided into 5 groups (n =18 each):sham group (group S),chronic constrictive injury group (group CCI),dexmedetomidine group (group D),ketamine group (group K) and dexmedetomidine + ketamine group (group DK).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 400 mg/kg.Neuropathic pain was induced by CCI in CCI,D,K and DK groups.The sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1mmintervals with 4-0 silk thread.In group S,the sciatic nerves were only exposed but not ligated.In D,K and DK groups,dexmedetomidine 50μg/kg,ketamine 10 mg/kg and dexmedetomidine 25μg/kg + ketamine 5 mg/kg were injected intraperitoneally,respectively,while the equal volume of normal saline was injected in S and CCI groups,once a day for 14 consecutive days after CCI.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CCI,and 3,7 and 14 days after CCI.Six animals were sacrificed after measurement of pain threshold at 3,7 and 14 days after CCI and the lumbar segments (L4-6) of the dorsal root ganglion were removed for determination of P2X4 R mRNA and P2X7 R mRNA expression by RT-PCR.Results Compared with group S,MWT and TWL were significantly decreased at 3,7 and 14 days after CCI in groups CCI,D,K and DK,the expression of P2X4R mRNA and P2X7R mRNA was up-regulated at 3,7 and 14 days after CCI in groups CCI,D and K,and the expression of P2X4 R mRNA and P2X7 R mRNA was up-regulated at 3 and 7 days after CCI in group DK (P ＜ 0.05).Compared with group CCI,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in groups D,K and DK (P ＜ 0.05).Compared with D and K groups,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in group DK (P ＜ 0.05).Conclusion The mechanism by which the combination of dexmedetomidine and small dose of ketamine produces a synergistic antinociception in rats with neuropathic pain may be related to down-regulation of the expression of P2X4 R mRNA and P2X7 R mRNA.

BACKGROUND: How to effectively harvest bone marrow mesenchymal stem cells with homogeneity is crucial for cartilage tissue engineering research. OBJECTIVE: To isolate and culture rat bone marrow mesenchymal stem cells and to observe biological characteristics of the cells. METHODS: Bone marrow mesenchymal stem cells were isolated from the femur and tibia of 30 Sprague-Dawley rats using the whole bone marrow adhesion method. Cell surface markers were identified using flow cytometry, cell proliferation ability was observed through cell counting, and cell survival rate was determined by MTT assay. RESULTS AND CONCLUSION: The harvested cells were mostly round and oval, which partially showed a triangular shape. Cell fusion appeared after 12-15 days of inoculation. The proliferation of passaged cells was accelerated. Two hours after passage, the cells evenly covered the bottom of the culture dish. With the increasing of cell generations, the cell proliferation ability was gradually decreased. The survival rates of cells at passages 1-5 were over 95%, significantly higher than those at passages 6 and 7 (P < 0.05). The harvested cells were negative for CD34, but positive for CD44. To conclude, the whole bone marrow adhesion method is effective to isolate rat bone marrow mesenchymal stem cells with stable growth and strong proliferation ability. Passage 5 cells can be selected as seed cells for cartilage tissue engineering.