Objective The adsorption effects of four sorts of dietary fibres from seaweeds on exogenous NO~-_(2) were studied in vitro and in vivo.Methods By simulating the environment of stomach and small intestine in vitro,and feeding rat in vivo.At the same time,the adsorption effect of wheat bran was studied comparatively.Results The results showed that:(1) the adsorption effect of dietary fibres on NO~-_(2) were related with the pH and the species of dietary fibres.Under the simulated stomach environment,the adsorption capacity of Laminaria japonica was the strongest with 1.19 mg?g~(-1),the next was Eucheuma with 0.94 mg?g~(-1),and Sargassum was the weakest with 0.60 mg?g~(-1).The adsorption ratio of Laminaria japonica and Eucheuma was significantly higher than that of wheat bran(P

Diabetic nephropathy is a complication of diabete and its incidence is increasing obviously these years. The pathogenic mechanisms of DN are complex, with multiple factors involved such as the glycometabolism disorders , hemodynamic changes, aldose reductase pathway activation, alterations in cytokines, oxidative stress, protein kinase C activation, lipid metabolism disorders and genetic susceptibility etc, which are not completely clear yet. Thus, the research progress of DN pathogenesis is to be reviewed.

Objective To observe the expression of leptin(Lep) receptor (LepR) and aquaporin 2 (AQP2) in the kidneys of obesity-related hypertensive rats ( OHR ) and to explore the mechanism of Lep resistance and water metabolic disorders in them.Methods OHR( model group) were induced by high-fat diet.Normal Wistar rats were chosen as normal control and hypertensive rats(SHR) as positive control.The serum level of triglycerides(TG), total cholesterol(TC), Lep, vasopressin ( AVP ) , angiotensinⅡ( AngⅡ) and β2-microglobulin (β2-MG ) was measured by enzyme linked immunosorbent assays ( ELISA) and renal morphology was observed by HE staining.The density of LepR and AngⅡtype 1( AT1) in the kidney was observed by immunohistochemistry.mRNA And protein expression of LepR and AQP2 in the kidney was assayed by real-time polymerase chain reaction and Western blotting.Results Compared with normal rats,the TG, TC, Lep, AVP, AngⅡand β2-MG of the model group were significantly increased (P<0.05), and protein and mRNA expression of LepR and AQP2 in the kidney were up-regulated (P<0.05).Compared with SHR group, TG, TC and Lep in serum of the model group were significantly increased (P<0.05).The concentrations of AVP,β2-MG and Lep was linearly related(R2 =0.87,R2 =0.95).Conclusion Water metabolic disorder and Lep resistance may be involved in the kidney injury of OHR, which may be one of the important pathogeneses of obesity-related hypertension.

AIM: To investigate the relationship betw een ADRP and the development of atherosclerosis. METHODS: Antisense oligodeoxynucleotide of mouse ADRP was constr ucted. The mouse peritoneum macrophages were cultured with Ox-LDL or Ox-LDL plus the antisense fragment. The cellular cholesterol was measured and the expressio n of ADRP was observed with RT-PCR and western blotting. New Zealand white rabbi ts were fed with high cholesterol chow for 12 weeks. The levels of serum lipid a nd cholesterol content of aortic wall were investigated. The areas of fatty stre ak of the aortas was measured after staining with Sudan Ⅳ. The aortic, and live r specimens with HE and immunohistochemistry staining were observed under light microscopes. RESULTS: Antisense oligodeoxynucleotides of mouse ADRP decreased cellular cholesterol ester, induced cellular lipid droplets and the expression of ADRP. The expression of ADRP was induced by high-cholesterol-diet feeding in rabbit atherosclerotic lesions. The fatty streak of the aorta with immunohistoch emistry staining was strongly positive for ADRP in animals fed with high cholest erol chow, and the liver was negative with or without high cholesterol chow. CONCLUSIONS: The expression of ADRP in vessel walls is related t o the atherosclerosis, and has a potential role in lipid accumulation in macroph ages and pathogenesis of atherosclerosis.

Objective To establish a method for the isolation and identification of platelets.Methods 10 healthy volunteers were selected to collect the EDTA anticoagulant venous blood of 3 tubes,each tube was 2 ml,which was divided into the whole blood cell tube,platelet rich plasma (control group),and stepped centrifugal platelet extract (experiment group).Platelet was isolated by simple centrifugation method(PRP) and stepped centrifugal method.The two groups were full blood count and analyzed by microscopic morphology and platelet activity test.Leukocyte specific HGB gene and platelet mitochondrial ND1 gene content was analyzed by real time PCR.Results Platelets were extracted and detected in control group and experimental group.Platelets were found and white blood cells and red blood cells were not remained in experimental group.Platelets and sporadic white blood cells were found in control group.The platelet pick up rate of experiment group was significantly higher than control group,the difference was statistically significant.Experimental gene content HGB of experiment group was significantly lower than control group,the difference was statistically significant (t=-3.281,-2.865,P＜0.05).ND1 gene content of experiment group higher than the control group,the difference was not statistically significant.There was no significant difference for platelet activity test between experimental group and control group (t=-0.046,-0.799,P＞ 0.05).Conclusion A isolation and identification method of stepped centrifugal platelet was established.The method can be used for the study of platelet gene and the functional analysis of platelets.

Objective Adsorption effect of dietary fibers from seaweeds on extrinsic estrogens was studied.Methods The methods included the tests in vivo and in vitro which simulated environments of stomach and small intestine of rat. Results The adsorption effect of four kinds of dietary fibers on estradiol in simulated environments of small intestine was better than that in simulated environments of stomach. The effect of Laminaria japonica was the best. And the rate of adsorption on estradiol was 108 g?g~ -1.It is 1.5,1.8, 3.1, and 3.0 times that of Sargassum,wheat blan,Gracilaria and Eucheuma,respectively. The rates of adsorption on estradiol of four kinds of dietary fibers from Gracilaria, Eucheuma, Sargassum and laminaria japonica was 11.1%,13.6%,27.5% and 38.7%, respectively. Adsorption rate of Sargassum and laminaria japonica was 1.9 and 1.3 times than that of wheat bran, respectiveiy which was 20.4%. While for Gracilaria and Eucheuma, adsorption rate was lower than that of wheat bran. Conclusion The dietary fibers from four seaweeds showed adsorption effect on extrinsic estrogens. The dietary fibers from brown seaweeds (Sargassum and laminaria japonica) had more adsorption effect than that of red seaweeds (Gracilaria and Eucheuma).

AIM: Based on the finding of adipophilin expression with the increase in cellular cholesterol, the aim of the present study was to look for the active site of adipophilin in cellular cholesteryl metabolism. METHODS: Mouse peritoneal macrophages were incubated with 80 mg/L Ox-LDL (Ox-LDL group) or 80 mg/L Ox-LDL plus 1 mmol/L adipophilin antisense oligonucleotides (Ox-LDL+antisense group), respectively. At the various time points, the incubated cell samples were observed with adipophilin immunofluorescence staining, flow cytometric analysis and cellular cholesterol analysis. RESULTS: The Ox-LDL+antisense group cells contained significantly lower cholesteryl ester (19.9?1.9) mg/g (protein) than that of cells in Ox-LDL group (46.6?3.4) mg/g (protein) at 4 days. From 12 h, expression of adipophilin in Ox-LDL group increased more quickly than that of the cells in Ox-LDL+antisense group. At day 4, the level of adipophilin expression in Ox-LDL group was significantly higher than that in Ox-LDL+antisense group. During the observation, the amount of Ox-r[CL-3H] LDL taking up increased gradually in both groups, however, from day 1 the taking up amount in Ox-LDL+antisense group was less than that in Ox-LDL group. There was a statistical difference between the two groups from day 2 to day 4. From 6 h to day 2, the relative ACAT activity increased in both groups. The relative ACAT activity kept unchanged from day 2 to day 4 in the two groups. At day 2, the relative ACAT activity in Ox-LDL+antisense group was significantly lower than that in Ox-LDL group. Correlative analysis between activity of ACAT and adipophilin expression showed than R2 were 0.6176 and 0.8212 (P

Objective To assess the validity of the Oxford classification for pediatric patients with primary IgA nephropathy (IgAN) and to analyze the correlations between clinical characteristics at time of biopsy and the Oxford classification,which identified four definitive histological features:mesangial hypereellularity,endocapillary proliferation,segmental sclerosis and tubular atrophy/interstitial fibrosis.Methods Clinical and pathological characteristics of 35 children with primary IgAN were analyzed.The scoring sheet was based on the Oxford classification of IgAN,and four pathological variables,namely mesangial hypercellularity (M),endocapillary proliferation (E),segmental sclerosis (S),and tubular atrophy/interstitial fibrosis (T) were assessed.A total of 35 children with IgAN were grouped according to the scores(M,E,S,T):the M0 and M1 group,E0 and E1 group,S0 and S1 group,T0 and T1/T2 group.These groups were compared in terms of estimated glomeralar filtration rate (eGFR),mean artery pressure (MAP) and proteinuria at time of biopsy.Results We found that the Oxford classification was significantly negatively correlated with eGFR (Pearson's correlation coefficient r =-0.48).However,the Oxford classification was shown to be positively associated with initial proteinuria per day(Pearson's correlation coefficient r =0.35).The M,E,S,T scores were strongly associated with proteinuria at biopsy (P ＜ 0.05),and the lesion S was not correlated with eGFR (P ＞ 0.05).The lesion T was significantly associated with eGFR (P =0.001) and MAP (P =0.03) at biopsy.Conclusion We confirmed that the Oxford classification of IgA nephropathy was valid for children.In addition,our study indicated that the four histological lesions M,E,S and T were significantly associated with clinical features.

Objective:The nutritional value and edible safety were evaluated for fastfood Eucheuma. Methods:The nutritional composition and the contents of heavy metal element and microbiology index were tested, and the test of acute toxicity and mutation were performed to evaluate the nutritional value and edible safety. Results:The main components of fastfood Eucheuma were dietary fibre and mineral components, and the contents of protein and fat were very low. The gross content of dietary fibre was 88.6%. The contents of Zn and Ca were the highest among the mineral components.The index of heavy metal elements and microbes(cfu) were lower than that of the national standard. It was non-toxic and non-mutagenic. Conclusion:The fastfood Eucheuma is a functional food, and also a safe food of new resource.

Objective To construct a recombinant Escherichia coli ( E. coli) with surface-dis-played lead specific binding protein PbrR and to further study intestinal colonization by the recombinant bac-teria in mice and gastrointestinal tolerance of the bacterial surface-displayed PbrR. Methods Chimeric pro-tein Lpp-OmpA coding sequence was chemically synthesized and inserted into the expression vector pET-21a to construct the outer membrane display vector pLOA. PbrR coding sequence was also obtained by chemical-ly synthesis and inserted into pLOA to generate the outer membrane display plasmid pLOA-pbrr. E. coli BL21 (DE3)pLysS was transformed with pLOA-pbrr and induced by IPTG. The expressed recombinant proteins were analyzed by 15% SDS-PAGE and Western blot assay. Lead adsorption capacity of the cell surface-dis-played PbrR in the simulated intestinal juice and tolerance of the recombinant E. coli to simulated gastric juice were analyzed, respectively. KM mice were orally given the induced recombinant bacteria by gastric lavage for 7 consecutive days and then were continually fed until day 30. The contents of recombinant bacte-ria in stool samples were detected by dilution plate method on day 7, 15 and 30. The recombinant protein with His tag was detected by immunoblotting on day 7 and 15. Results Based on Lpp-OmpA, the PbrR outer membrane display vector was successfully constructed. The recombinant fusion protein Lpp-OmpA-PbrR-His tag was highly expressed in E. coli. The recombinant E. coli strains displaying PbrR on their outer membrane accumulated a significant level of Pb2+ in simulated intestinal juice. Moreover, those strains showed a tolerance to gastric acid in vitro and could colonize in the intestinal tracts of mice via oral infection. The surface-displayed recombinant fusion protein showed a better tolerance to the environment of digestive tract. Conclusion The recombinant E. coli strain displaying PbrR on its surface showed a stronger capabili-ty of lead accumulation from simulated intestinal environment and could colonize in the intestinal tracts of mice. The surface-displayed recombinant PbrR also showed a good tolerance to digestive juice. This study paved the way for further researches on the selective elimination of lead by biosorption based on animal mod-els.