Objective To investigate the regulation of blood-testis barrier by Rab13-PKA pathway in rats.Meth-od First, shRNA vector targeting at Rab13 was constructed and then the Rab13 shRNA was transfected into the rat testis by injection.Western blot was used to detect the knock-down effect of Rab13 and the expression of blood-testis barrier ( BTB) constituent proteins.PKA activity was detected by autoradiography and scintillation counting.Further, immunoflu-orescence analysis and phalloidin staining were applied to observe the distribution of occludin and F-actin, respectively. Results The expression level of Rab13 in the testis was reduced by approximately 70%after transfection of Rab13 shRNA as compared with the non-targeted control group ( P<0.01 ) , while the expression of BTB constituent proteins remained unchanged.PKA activity was significantly increased after Rab13 RNAi transfection (P<0.01).The distribution of occlu-din at BTB was remarkably increased after Rab13 RNAi silencing around stage VIII but not at other stages of the seminifer-ous epithelial cycle.The assembly of F-actin at BTB was also intensified in Rab13-silenced testis.Both the changes of dis-tribution of occludin and F-actin induced by Rab13 shRNA were found to be antagonized by the PKA specific inhibitor H89.Conclusions Rab13 can modulate the distribution of occludin and F-actin at the blood-testis barrier in rats by regu-lating PKA activity, which may participate in the regulation of BTB function.

OBJECTIVE:To explore the trend of cooper enrichment in hirudinidae influenced by culture environment. MET-HODS:The soil containing low-content,medium-content and high-content of cooper groups(30.00,60.00,90.00 μg/g)and water culture control group were set up. Hirudinidae leech were culture for 60 d,and sampled every 15 days. ICP-MS techniques was used to determined and compared the contents of cooper in W. pigra and soil. RESULTS:In first 15 days,the contents of cooper in leech from 4 groups increased greatly,compared with before;in the following 15 days,the content of cooper kept stable in high-content group while decreased in other 3 groups;in the 30-45 day,the contents of cooper increased rapidly in 4 groups,and those of low-content group and high-content group reached the peak in this experiment;in the last 15 days,the contents of cooper in control group and high-content group increased continuously,while those of low-content and medium-content groups decreased to some extent. Compared with before,the contents of cooper in leech from control group,low-content,medium-content and high-content groups increased by 292%,186%,293%,464% respectively;those of soil from latter 3 groups increased by 81.12%,35.98% and 21.28% respectively. CONCLUSIONS:The content of cooper in leech increase with time,and is positively correlated with the content of cooper in soil. It is suggested to control the content of cooper in hirudinidae through controlling cul-ture environment when hirudinidae are cultured as medicinal material,in order to meet the quality standard of heavy metal in medic-inal material.

This study was aimed to detect the heavy metal residues of medicinal leeches, in order to understand the current market circulation situation of medicinal leeches, and to analyze possible factors which may cause heavy metal pollution, and to provide references for the standardized safe cultivation of medicinal leeches. Inductively coupled plasma mass spectrometry (ICP-MS) technique was applied to detect heavy metal residues in medicinal leeches. The results showed that medicinal leech samples tested for heavy metal content severely exceeded the standard, which caused a great threat to the safety of medication. It was conclude that more attentions should be paid on factors for causing heavy metal accumulation within medicinal leeches during the breeding process. The related department should also list safety limits explicitly and separately for animal drugs such as medicinal leech during the development of quality standards of Chinese materia medica.

In this study cationic and heparinized polyurethanes (PUs) were synthesized by a two-step solution polymerized method. Cationic and heparinized PUs were investigated by infrared spectroscopy, electron spectroscopy for chemical analysis (ESCA) and turbidity method. At the same time, the PUs proved of good biocompatibility through the laboratory tests, including blood coagulation time (CT), activated partial thromb plastin time (APTT) and fibroblast culture. These materials have good biocompatible function.
Blood Coagulation Tests ; Coated Materials, Biocompatible ; chemistry ; Heparin ; pharmacology ; Humans ; Materials Testing ; Partial Thromboplastin Time ; Polyurethanes ; chemical synthesis ; chemistry ; Surface Properties

BACKGROUND:Reports have demonstrated that cytotoxicity produced by ferric oxide (Fe2O3) nanoparticles is associated with cellular lipid peroxidation. Whether Fe2O3 nanoparticles have toxicity to macrophages, and what is the association of toxic mechanism and oxidization?OBJECTIVE: To observe the effects of different concentrations of Fe2O3 nanoparticles on the oxidative damage of macrophages.DESIGN: A controlled observation experiment.SETTING: School of Public Health, Southeast University.MATERIALS: RAW264.7 cells were peritoneal macrophages of mouse and purchased from Shanghai Institute of cells, Chinese Academy of Sciences. Fe2O3 nanoparticles (30 nm) suspension was provided by Department of Biomedical Engineering, Southeast University). Fe2O3 nanoparticle suspension was placed in 60 ℃ water for 10 hours,then in 37 ℃ water overnight. This procedure was repeated 3 times for germicidal treatment. Then, the suspension was packed into small bottles and stored at 4 ℃ for later use. DMEM high glucose culture fluid (Gibco Company,USA); trypsinase (Difco Company, USA, imported); new-bom calf serum(Sijiqing Company, Hangzhou); hydrogen dioxide (H2O2, Gibco Company); Kits for measuring hydrogen dioxide(H2O2), hydroxy radical (·OH), superoxide anion radical (O2·-), lactic acid dehydrogenase, ultramicro ATP enzyme and Coomassie brilliant blue protein levels (Jiancheng Biotechnique Co., Ltd.,Nanjing).METHODS: This experiment was carried out in the laboratory of Department of Labor and Environmental Health, School of Public Health, Dongnan University between March 2006 and July 2006. RAW264.7 cells (Abelson murine leukemia virus-induced tumor) were cultured in DMEM (Gibco Company) containing 100 g/L fetal bovineserum, 100 000 U/L penicillin and 100 mg/L streptomycin in the environment of 5% CO2. Cell growth was observed under an inverted radical in the cells: 1.5×108 L-1 macrophages were inoculated to 24-well plate, 1 mLa well. After the macrophages were cultured for 24 hours in incubation at 37 ℃ in a humidified atmosphere containing 5% CO2. 1.070 0, 0.5350 and 0.2675 g/L Fe2O3 nanoparticles (30 nm) suspension-intervened macrophages were set as Fe2O3 nanoparticle group, and normal saline group was set as control group. Following the intervention of nanoparticles, macrophages were disrupted with Determination of the activities of lactate dehydrogenase (LDH), Na+-K+-ATPase and Ca2+-Mg2+-ATPase: Macrophages in the Fe2O3 nanoparticle group and control group were treated as above. The activities of LDH in culture medium were determined according to the instruction of reagent kit (Nanjing Jiancheng Bioengineering Co., Ltd). And the activities of Na+-K+-ATPase and Ca+-Mg2+-ATPase were also determined according to the instruction of reagent kit (Nanjing Jiancheng Bioengineering Co., Ltd) at low temperature. MAIN OUTCOME MEASURES: ①Effects of different concentrations of Fe2O3 nanoparticles on the production of H2O2, ·OH and O2·- in RAW264.7 cells.②Effects of different concentrations of Fe2O3 nanoparticles on the activities of LDH ,Na+-K+-ATPase and Ca2+-Mg2+-ATPase in RAW264.7 cell culture fluid.RESULTS: ① Level of ·OH free radical in Fe2O3 nanoparticle 0.267 5, 0.535 0, 1.070 0 g/L groups was higher than that in control group, respectively [(0.605±0.066), (0.410±0.080), (0.764±0.051), (0.285±0.057)mkat/g, P ＜ 0.05]; Level of respectively [(9.935±1.159), (8.912±0.131), (13.479±0.752), (5.635±0.475)μkat/g,P ＜ 0.05]; Level of H2O2 in Fe2O3 nanoparticle 1.070 0 g/L group was higher than that in the control group [(14.695±2.815), (2.397±0.399) mmol/L, P ＜increased (P ＜ 0.05). Fe2O3 nanoparticles had effects on the activities of Na+,K+-ATPase and Ca2+,Mg2+-ATPase. With the increase of dose of Fe2O3 nanoparticles, the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase were gradually decreased. There were significant differences as compared with control group (P ＜ 0.05)CONCLUSION:Increasing dose of Fe2O3 nanoparticles wouldcause more H2O2,·OH and O2·- free radicals in the cells, increase cell membrane permeability and inhibit the activities of LDH, Na+-K+-ATPase and Ca2+-Mg2+-ATPase.

Objective To prepare a kind of Gelsolin-targeted ultrasound contrast agent (GSN-PLGA) and to explore its targeting and imaging effection in vitro.Methods The high molecular PLGA-COOH ultrasound contrast agents were prepared by a modified double emulsion technique and then conjugated with Gelsolin monoclonal antibody by carbodiimide technique.The physical property of contrast agent was determined.And the connectivity condition of ultrasound contrast agent with Gelsolin monoclonal antibody was estimated.The targeting ability and the effect of enhancing ultrasound imaging in vitro were explored.Results The average diameter of GSN-PLGA was (575.67 ± 4.71) nm.The potential was (-11.46±1.19) mV.And the binding rate of Gelsolin monoclonal antibody was 96.93％.In vitro experimentshowed more GSN-PLGA could be intaked by Hca-F cells and the ultrasound imaging cloud be enhanced greatly.Conclusion The GSN-PLGA nanoparticle can bind to Hca-F cells specifically and can enhance the ultrasound imaging greatly.

It has been previously thought that calcification is a feature of the stability of atherosclerotic plaques. However, recent studies have shown that microcalcification in atherosclerotic plaques is significantly associated with plaque vulnerability. The relationship between atherosclerotic plaques and calcification is unclear, and the specific role of calcification in atherosclerotic plaques remains controversial.

Objective To measure the number of lymphocytes, B lymphocytes, CD5+B lymphocytes and level of IL-10 in peripheral blood of patients with systemic lupus erythematosus (SLE), and analyze their effects in the disease. Methods In this study, 84 cases of patients with SLE were randomly selected and evaluated according to the activity index (SLEDAI). These cases were divided into low activity group (SLEDAI<9) and high activity group (SLEDAI≥9). Ten healthy individuals were selected as the control group at the same time. The number of peripheral blood lymphocytes, B lymphocytes, CD5 + B lymphocytes, erythrocyte sedimentation rate (ESR), C3, C4 and interleukin (IL)-10 levels in serum were measured respectively and the correlation between the above indexes and SLEDAI and complement levels were analyzed. Pair-wise comparison of means of groups was conducted with one-way ANOVA. Comparison between the two groups was conducted by LSD-t test. Correlations between variables were carried out using Spearman's rank correlation test. Results The total number of lymphocytes in SLE group was lower than that in normal control group ( F=7.216, P<0.001); The number of CD19+ B lymphocytes in SLE group was higher than that in normal control group (F=3.589, P=0.036). The number of CD5+B lymphocytes of peripheral blood [(2.5±0.6)%] in patients with systemic lupus erythematosus was significantly lower than that in the normal control group [(3.2 ±0.8)%], but the difference was not statistically significant (t=3.412, P=0.698). The number of CD5+B lymphocytes in the high activity group was significantly lower than that in the low activity group (t=7.365, P=0.027)and the normal control group (t=5.649, P=0.002). The number of CD5+ B lymphocytes was negatively correlated with SLEDAI score (r=-0.692, P=0.001) and positively associated with the level of complement 3 (r=0.305, P=0.038), but not with complement 4 and ESR (P>0.05). In addition, the level of serum IL-10 in whether the low activity group (t=1.935, P=0.031) or the high activity group (t=3.048, P=0.012) was all higher than the normal control group. The level of serum IL-10 in patients with systemic lupus erythematosus was positively associated with SLEDAI score (r=0.425, P=0.024) and ESR (r=0.479, P=0.008), but was negatively correlated with complement 4 (r=-0.359, P=0.031). Conclusion The total number of lymphocytes in patients with SLE decreases significantly, while B lymphocytes increases significantly. The number of CD5+ B lymphocytes and the serum IL-10 level are also changed. It maybe related to the patient's inflammatory environment, and the number of CD5+B lymphocytes and the serum IL-10 level may be associated with disease activity.

Objective:To observe any stimulatory effect of intermittent theta burst stimulation (iTBS) on the cerebral swallowing cortex and the cerebellar swallowing motor area and to explore the related mechanisms.Methods:Forty-four healthy right-handed subjects were divided at random into a dominant cerebellum group ( n=15), a non-dominant cerebellum group ( n=15) and a control group ( n=14). In the dominant cerebellum group, iTBS was administered to the cerebellum of the dominant hemisphere, and the other hemisphere was given sham stimulation. In the non-dominant cerebellum group, it was the opposite. The dominant cerebellum received the sham stimulation. In the control group both hemispheres received sham stimulation. Before and after the stimulation, single-pulse transcranial magnetic stimulation (TMS) was applied to the representative regions of suprahyoid muscles in bilateral brain and cerebellum to observe changes of the latency and amplitude of motor evoked potentials (MEPs). Results:After the intervention the MEP amplitude of the bilateral swallowing cortex and the stimulated cerebellum had increased in the non-dominant cerebellum group, with increased MEP amplitude only from the stimulated cerebellum of the dominant cerebellum group. Compared with the control group, the non-dominant cerebellum group showed the greatest improvement in MEP amplitude of the stimulated bilateral cerebral cortex and cerebellum. Improvement in the dominant cerebellum group was significantly smaller. However, there were no significant differences in MEP latency or the percentage change in MEP latency from baseline among the three groups.Conclusions:Applying iTBS to either the non-dominant or the dominant cerebellum excites the brain areas related to swallowing, but in different ways.

Objective To investigate the structure of deoxyhypusine synthase(DHS)in Saccharomyces cerevisiae(Dys1)and unravel the molecular mechanism of hypusine lysine modification,providing a theoretical basis for the treatment of highly proliferative diseases such as human immunodeficiency virus type 1(HIV-1)replication.Meth-ods Using the E.coli BL21 expression system,an in vitro expression vector was constructed and used to express the protein of Dys1.Dys1 protein samples were purified using methods such as affinity chromatography and molecu-lar sieving to achieve protein purification and isolation.The crystals of Dys1 were obtained using the crystallized so-lution containing 6％Polyethylene Glycol(PEG)8000,0.1 mol/L N-2-hydroxyethylpiperazine-N-ethane-sulphoni-cacid(Hepes)pH 6.5,and 8％ethylene glycol.The crystal structure of Dys1 was resolved at a resolution of 2.8 ? using X-ray crystallography.The structural analysis was performed with CCP4i and Coot software.Results The overall structure of Dys1 was a tetramer,each monomer containing a catalytic site and a cofactor NAD+binding site.The core region of the monomer adopted a Rossmann fold.The amino acid residues involved in the substrate binding sites were highly conserved among eukaryotes.Conclusion The crystal structure of Dys1 is being resolved for the first time.It reveals the binding mode of the cofactor NAD+to the enzyme and confirms that the enzyme functions as a tetramer,with the N-terminus serving as an essential modulator for its catalytic activity.