Objective To explore the effects of different extraction methods on yield of Smilax glabra polysaccharides and antioxidant activities. Methods Crude polysaccharides were extracted by using cold water, hot water, NaOH aqueous solution, and EDTA?2Na aqueous solution . Antioxidant activities of Smilax glabra polysaccharides were compared by the methods of scavenging superoxide anion radical , scavenging hydroxyl radical or reducing power. Results Yield of crude cold water-soluble polysaccharides, crude boiling water-soluble polysaccharides, crude alkali-soluble polysaccharides and crude acid-soluble polysaccharides were 0.31%, 1.1%, 10.8 %, 2.0%, respectively. Polysaccharides by using four kinds of different extraction methods had strong scavenging superoxide anion radical power; crude cold water-soluble polysaccharides, crude boiling water-soluble polysaccharides , crude acid-soluble polysaccharides had strong scavenging hydroxyl radical and reducing power , the capacity increased with the increasing concentration of Smilax glabra polysaccharides; the scavenging hydroxyl radical and reducing power capacity of crude alkali-soluble polysaccharides is very poor. Conclusion Polysaccharides by using four kinds of different extraction methods have certain antioxidant activity, the antioxidant capacity to scavenge free radical in vitro showed positive correlation with the concentration of Smilax glabra polysaccharides, considering the yield of polysaccharides, EDTA?2Na aqueous solution extraction method is the best.

Objective To identify the distribution feature of methylenetetrahydrofolate reductase (M T HFR) gene polymorphism of Buyi ,Dong ,Miao nationality in Guizhou .Methods The MTHFR(677 and 1 298) genotypes of Buyi ,Miao and Dong healthy indi-viduals were determined by TaqMan-MGB probe genotyping method and constructed haplotypes .Results There were significant difference of MTHFR 677C/T genotype and allele frequencies among 3 groups(P<0 .05) ,There was significant difference of geno-type between Buyi and Miao nationality ,and there were significant differences of genotype frequencies in Buyi nationality and Dong and Miao nationality(P<0 .01) .There were no differences of MTHFR 1298A/C genotype frequencies among Buyi ,Dong and Miao nationality(P> 0 .05) .Buyi nationality had the lowest frequency in double wild homozygous type (677CC/1298AA) ,677TT/1298CC double mutation homozygous and 677TT/1298AC combination in above three minorities was not found .There were linkage disequilibrium between 677C/T and 1298A/C in Buyi and Miao nationality .Conclusion The genotypes frequencies of MTHFR 677T T/1298AC are significant differences among different regions and different ethnic groups .

Objective To investigate the roles of microRNA-382 (miR-382) in the pathogenesis of renal tubulointerstitial fibrosis (TIF).Methods Human kidney epithelial cells (HK2)transfected with miR-382 inhibitor (antagomiR-382) were used to examine the effect of miR-382 abundance on cell polarity,as well as to test the complementary relationship between miR-382 and its predicted target gene heat shock protein 60 (HSPD1),which was further verified by 3'-untranslated region luciferase assay and site-directed mutagenesis.The role of miR-382 played in the development of renal interstitial fibrosis and redox regulation was examined in a mouse unilateral ureteral obstruction (UUO) model.Locked nucleic acid (LAN)-modified anti-miR-382 was intravenous delivered via tail vein 30 min prior to UUO,and repeated the dosage 24 h after the surgery.For clinical verification,renal biopsy specimens from 12 IgA nephropathy (IgAN) patients were collected,6 patients with moderate to severe TIF and 6 patients without TIF.The relative abundance of miR-382 and HSPD1 protein was analyzed by using in situ hybridization and immunohistochemistry.Results HSPD1 was confirmed to be a new,direct target gene of miR-382 by in vitro 3'-untranslated region luciferase assay and sitedirected mutagenesis.The development of epithelial transition in HK2 cells was accompanied with upregulation of miR-382 [(6.54±0.96) vs (1.12±0.26),P ＜ 0.05].Blocking the expression of miR-382 could reversed the progression of epithelial transition partially.In UUO mice the abundance of miR-382 was up-regulated [(6.89 ± 2.47) vs (1.00±0.42),P ＜ 0.01] while HSPD1 and Trx were downregulated compared with the sham group.Down-regulation of miR-382 was associated with significant decrease in TIF,but increase in HSPD1 and thioredoxin protein compared with UUO group [HSPD1:(0.34±0.10) vs (0.14±0.05);Trx:(0.79±0.18) vs (0.36±0.16);all P ＜ 0.05].The expression of miR-382 was up-regulated and HSPD1 was significantly down-regulated in IgAN patients with TIF.Conclusions miR-382 play an important role in renal tubulointerstitial fibrosis in human and mice.HSPD1 is one of the target genes of miR-382.The down-regulation of HSPD1 and the decrease ability of anti-oxidative stress may be the important mechanism of miR-382 involved in renal tubulointerstitial fibrosis.

Objective To understand how Vibrio vulnificus hemolysin (VvhA) affects the viability of murine liver CD4+ T cells as well as its effects on the numbers of mitochondria and the expression of CD62L. Methods The primary murine liver monocytes (MNs) were isolated from C57BL/ 6 mice and then treated with recombinant VvhA (rVvhA) for 6 hours in vitro. The viability of murine liver CD4+T cells and the expression of CD62L were measured by staining with anti-mouse CD4, CD8, CD44, CD62L and cell via-bility fluorescent dye or fluorescent antibody. Moreover, the cells were simply incubated with MitoTracker or JC-1 probes to label mitochondria and mitochondrial membrane potential, which were further analyzed by using flow cytometry analysis. Results With the increase in the doses of rVvhA, the viability of murine liv-er CD4+T cells was decreased from 81. 5% to 15. 8% . The expression of CD62L on the surface of murine liver CD4+T cells was dramatically decreased. Both the murine liver na?ve and effector CD4+ T cells were sensitive to the cytotoxicity of rVvhA. Moreover, treating murine liver CD4+ T cells with rVvhA resulted in significantly decreased numbers of mitochondria and lower mitochondrial membrane potential. Conclusion The cytotoxicity of rVvhA to murine liver CD4+T cells might be achieved through inhibiting the expression of CD62L, decreasing the numbers of mitochondria and lowering mitochondrial membrane potential.

Objective To investigate the mechanism of BVT. 2733 on insulin resistance, by using diet-induced obese (DIO) mice model. Methods After having been balanced for 3 days, the C57BL/6J mice were randomly divided into a normal diet group and a high-fat diet (HFD) group. After 20 weeks, the obese mice were further randomly divided into an obese control group, a BVT. 2733 group and a pioglltazone (PGZ) group and they were orally administered with placebo, BVT. 2733 and PGZ separately for two weeks.Adiponectin and leptin mRNA expression levels from adipose tissue were analyzed with real-time quantitative PCR. The levels of plasma glucose, serum insulin and adiponectin were measured with biochemical technology, radioimmunoassay and ELISA. Adipocyte sizes were observed with immunohistocbemistry.Results The body weight, plasma glucose and serum insulin levels raised(P<0.05)in the HFD group and the adipocyte sizes were bigger. Serum insulin levels significantly reduced (P<0.05) and adipocyte sizes reduced, while plasma adiponectin level raised (P<0.01)in the two treatment groups as compared with those in obese controls. Both the mRNA expressions of adiponectin and leptin upregulated(P<0.05)in the PGZ group, but their expressions in the BVT. 2733 group did not alter significantly. The body weight of the mice reduced significantly in the BVT. 2733 group. Conclusion BVT. 2733 can reduce body weight significantly and improve insulin resistance, but cannot influence the expression of adipocytokines.

Objective To study the techniques of the treatment for epidermoid with endoscope-as- sisted microneurosurgery.Methods The suboccipital,infratemporal transtentorial approach and endoscope- assisted microneurosurgery were used.Results Total resection was achieved in 10 cases,and subtotal resec- tion was made only in 2, and had no complications of all.Conclusion Endoscope-assisted microneuro- surgery can increase the total-resection rate for tumors,and reduce complications.

BACKGROUNDProbucol is known to reduce the development of atherosclerotic lesions, but its impact on vascular remodeling associated with de novo atherosclerosis is incompletely understood. We therefore examined the effect of probucol on vascular remodeling in a rabbit model of established atherosclerosis.

METHODSAortic atherosclerosis was induced by a combination of endothelial injury and 10 weeks' atherogenic diet. Animals were then randomized to receive the foregoing diet without or with 1% (wt/wt) probucol for 16 weeks. At the end of week 26, in vivo intravascular ultrasound, pathological, immunohistochemical and gene expression studies were performed.

RESULTSProbucol significantly decreased vessel cross-sectional area, plaque area and plaque burden without effect on lumen area. More negative remodeling and less positive remodeling occurred in the abdominal aortas of probucol group than the control group (56% vs. 21%, 18% vs. 54%, respectively, both P < 0.01). In addition, the probucol group showed a smaller mean remodeling index relative to the control group (0.93 ± 0.13 vs. 1.05 ± 0.16, P < 0.01). Furthermore, probucol treatment decreased macrophage infiltration, inhibited apoptosis of cells within plaques, and reduced the production of matrix metalloproteinases-2, -9, cathepsin K and cathepsin S (all P < 0.01).

CONCLUSIONSThese findings suggest that probucol may attenuate the enlargement of atherosclerotic vessel walls and be associated with a negative remodeling pattern without affecting the lumen size. This effect may involve inhibition of extracellular matrix degradation and prevention of apoptosis in atherosclerotic plaques.

Animals ; Anticholesteremic Agents ; pharmacology ; Aorta ; pathology ; Apoptosis ; drug effects ; Atherosclerosis ; drug therapy ; metabolism ; pathology ; Lipids ; blood ; Macrophages ; drug effects ; physiology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Oxidative Stress ; Probucol ; pharmacology ; Rabbits ; Ultrasonography, Interventional ; methods

OBJECTIVETo investigate the effect of advanced glycosylation end products (AGE) on cell cycle of epidermal keratinocyte and its possible signal pathway.

METHODS150 mg/L AGE-human serum albumin (AGE-HSA) was prepared in vitro. Primary cultured keratinocytes in logarithmic growth phase were harvested and divided randomly into: A group [with treatment of defined keratinocyte-SFM (DK-SFM) serum-free medium], B group (with treatment of DK-SFM medium including 150 mg/L AGE-HSA), C group (with DK-SFM medium after treatment of U0126) and group D (with D K-SFM medium including 150 mg/L AGE-HSA after treatment of U0126). Cell cycle distributions were analyzed by flow cytometer. The protein levels of cyclin D1, cyclin B1, CDK4 and p44/42 MAPK were measured by Western blot.

RESULTSCompared with those of A group, the percentage of S-phase and G2/M-phase keratinocytes were decreased obviously in B group, the percentages of G2/M -phase keratinocytes showed the same tendency in C and D groups [(9.7 +/- 1.1)% , (9.8 +/- 0.7)%, respectively, P <0.05]. Compared with that of A group, the expression of cyclin D1 were decreased significantly in other groups, among which a weak expression was showed in D group. There was no obvious difference between A and B groups in CDK4, or cyclin B1 and p44/42 MAPK protein levels ,which were significantly higher than those in C and D groups.

CONCLUSIONAGEs inhibit the progress of cell cycle of keratinocytes by downregulation of cyclin D1 expression.

Animals ; Cell Cycle ; Cyclin D1 ; metabolism ; Epidermis ; cytology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Glycation End Products, Advanced ; metabolism ; pharmacology ; Keratinocytes ; cytology ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

Background Glaucoma is one of the leading causes of blindness worldwide;while the different types of glaucoma is vary from different region. Objective Present study was to survey the prevalence and types of glaucoma among peasants of Uigur adults in Kuche county and offer the basis for the prevent and treatment of glaucoma in Uigur nationanlity. Methods 4191 Uigur peasants aged 40 years or above were collected in Kuche county for the survey of prevalence and types of glaucoma by randomized cluster sampling in March and April of 2009. The subjects were grouped into 40-,50-,60- and ≥70 years groups according to the distribution of age. The disease history of glaucoma, regular eye examination, funds examination and measurement of the anterior chamber depth, gonioscope were performed in the all subjects. Darkroom prone test and mydriasis test were carried out in suspicious glaucomous patients. The depth of periphery anterior chamber was assessed based on van Herick' s criteria, and the width of chamber angle was graded based on Scheie' s method. The standardized training was performed. This survey approved by Xinjiang Medical Ethics Committee, all subjects signed the informed consent before the examination. Results 4191 of 4873 subjects finished all the examinations with the response rate 86%. All the subjects showed a good compliance. The prevalence of glaucoma was 3. 79% , and the prevalence of primary angle-closed glaucoma(PACG) .primary angle-open glaucoma (PAOG) and secondary glaucoma was 2.22% ,0.26% and 1. 31% respectively, showing a significant difference among the different types of glaucoma( P<0. 05). The prevalence of glaucoma was elevated with aging (χ2 - 116. 69 ,P<0. 05) and presented a high rate in male subjects compared with female ones(χ2 = 7. 34, P<0. 05 ). Bilateral blindness was found in 19.75% glaucoma peasants, in which 25.3% glaucoma peasants received anti-glaucoma surgery. The distribution of visual acuity of patients was of significant difference among different age groups(χ2 = 37. 69 ,P<0. 05 ) . Conclusions The prevalence of the glaucoma among Uigur peasants in Kuche county was higher than most area no matter inland or overseas. PACG still is the common type in those people.

OBJECTIVETo evaluate the effect and complication of surgical treatment for Pilon fracture using the posterolateral approach.

METHODSFrom August 2009 to March 2011, 15 patients with Pilon fractures (2 in B3,13 in C) and with a separate displaced posterior malleolar fragment was treated in two-stage: the first stage management was on stabiliztion of the soft tissue envelope with temporary external fixator of spanning arthritis, and the second stage management was open reduction and internal fixation with posterolateral approach and anteromedial or anteralateral approach.

RESULTSAll patients were followed-up for 12 to 17 months (14.2 months in average). Thirteen of the 15 fractures healed, but 2 fractures needed autologous bone graft procedure duo to nonuion. There was no wound complication related to poterolateral incion. Fourteen fractures had less than 2 mm of incongruity of distal tibia joint. According to Baired-Jackson criteria, the results were excellent in 2 cases, good in 7, fair in 4, and poor in 2.

CONCLUSIONThe posterolateral approach offers direct visualization for the reduction and fixation of the fibula and posterior distal fragment of the tibia Pilon fractures, faciliate the management of this difficult fracture pattern.

Adult ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; surgery