Objective To study the relationship between fucoidans structure and their antioxidant activities. Method Fucoidans (HF) were extracted from Laminaria japonica and was separated by anion exchange chromatography on DEAE-Cellulose 52 column (2.5?40 cm), and two fractions HF1 and HF2 were obtained. Fucoidans of low molecular weight (LF) was made after hydrolysis of HF by hydrochloric acid and purification by ultrafiltration. Chemical methods and gas chromatography were used to analyse the sugar composition of these polysaccharides; and infrared spectrum was utilized to determine their structure. The activity of inhibiting hydroxyl radical (?O H ) was evaluated by means of Fenton reaction. Results In HF1, 18.834% uronic acid, 44.197% L-fucose, 31.193% D-galactose and 24.612% D-mannose were detected. As to HF2, 5.878% uronic acid, 81.119% L-fucose and 18.881% D-mannose were found. L-fucose content in LF was 13.033%, equal to 32.084% of that in HF 40.621%. Both in HF1 and LF, sulfate groups were at the C-2 and C-4 positions of L-fucose, while in HF2, sulfate groups only at C-4 position. The concentration of fucoidans inhibiting 50% ?O H (IC50) was 7.6, 5.2 and 8.9 mg/ml for HF, HF1 and HF2 respectively. Nevertheless, LF approximately lost this activity. Conclusion The molecular weight, content and composition of sugar and the combining sites of sulfate groups were found to be related with the activity of inhibiting ?O H radical in fucoidans.

Objective To study the clinical characteristics,treatment and prognosis of neonatal VACTERL association.Method The clinical data of newborns diagnosed with VACTERL association from January 2010 to December 2015 were collected and retrospectively analyzed.Result A total of 33 patients diagnosed with VACTERL association were included,including 23 males and 10 females.Among them,17 cases were term infants,15 cases premature infants and 1 case of overdue birth,with an admission age of 1 to 24 days.The most common deformities were cardiac anomalies (C) in 27 cases (81.8％),followed by anal atresia/anorectal malformation (A) in 25 cases (75.8％),renal deformity (R) in 24 cases (72.7％),limb abnormalities (L) in 20 cases (60.6％),Tracheoesophageal fistula (TEF) in 8 cases (24.2％) and vertebral abnormalities (V) in 3 cases (9.1％).11 cases (33.3％) had other deformities.Among these 33 patients,24 cases had 3 types of malformations and 9 cases had 4 types of malformations.The most common combination was ACR (n =8).20 patients had no abnormalites on chromosome karyotype test including 2 patients had normal gene microarray results.16 patients received surgical treatment during neonatal period and 13 of them recovered and discharged.Among the other 17 cases received no surgery,only 1 patient improved and discharged.A telephone follow-up was proceeded in 14 discharged cases at 1 year old.Among them,13 cases had good prognosis,however,the remaining one was dead.Conclusion VACTERL association is a rare non-random combination of multiple malformations.The early discovery and appropriately treatment after diagnosis will improve the prognosis and prevent death.Doctors should reinforce the ability to detect various types of deformities and examine the chromosome and gene properly.

Objective To evaluate the alteration of CD4+ regulatory T cells in peripheral blood from patients with ankylosing spondylitis(AS). Methods Seventy-eight AS patients and 50 healthy individuals were included in this study. The proportion of CD4+CD25+CD127lo/- T cell population in CD4+ T cells as well as that cytotoxic T lymphocytes(CTL) and NK cells in lymphocytes was evaluated by flow cytometry. Serum TGF-β and TNF-α were measured by ELISA. The inhibitory function of CD4+CD25+ regulatory T cells was measured by mixed lymphocyte culture. Results The population of CD4+CD25+CD127lo/- T cells in peripheral blood of AS patients accounted for (4.36±1.21)% of CD4+ T lymphocytes, which was significantly lower than that in healthy individuals (P<0.05). The CD4+CD25+CD127lo/- T cells population in AS patients was positively correlated with TGF-β level, but negatively with TNF-α. Compared with healthy individuals, the function of CD4+CD25+ regulatory T cells in the inhibition of alloreactive T cells was lower in AS patients, which was related to the decreased secretion of TGF-β. Conclusion The CD4+ regulatory T cells in peripheral blood of AS patients are significantly decreased and its function is defective, which leads to immune regulatory dysfunction in vivo. It may be one of immune pathogenesis mechanisms of AS.

Background Mutant C57BL/6 mouse with corneal opacity (B6-Co) appears eye open at birth (EOB) phenotype,which is a good animal model in the study of developmental mechanism of eyelid.Investigating the relationship between serum response factor (SRF) and EOB phenotype can provide theoretical support for the research on the mechanism of innate defects in eyelid development in humans.Objective This study was to assess the dynamic expressions of SRF in eyelid of embryonic B6-Co mouse.Methods Total RNA was extracted from B6 and B6-Co mice eyelid tissue at embryonic day 16.5 (E16.5 d),E17.5 d and E18.5 d.The relative expression levels of SRF mRNA and protein in the eyelid tissue of B6 and B6-Co embryonic mice were assayed by real-time quantitative PCR and Western blot,respectively.In situ expressions of SRF protein in eyelid of B6-Co mice and B6 mice were detected using immunofluorescence technique.The use and care of the animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals of Nantong University.Results The relative expression levels ofSRF mRNA in the eyelids were 0.41±0.06 and 0.24±0.17 in E16.5 d and E17.5 d of B6-Co mice,showing a significant decline in comparison with 1.03 ±0.17 and 1.01 ±0.09 in the B6 mice (P =0.025,0.017).The expression levels of SRF protein in the eyelids of E16.5 d and E17.5 d B6-Co mice were 0.08±0.01 and 0.08± 0.01,which were significantly lower than 0.12 ±0.03 and 0.13 ± 0.02 of B6 mice (P =0.036,0.024).However,there were no significant differences in the expression levels of SRF mRNA and protein in E18.5 d between the B6-Co mice and B6 mice (P =0.387,0.774).Immunofluorescence assay displayed that SRF was expressed in the keratinocytes of eyelids in both mice,but the fluorescence intensity was weaker in the B6-Co mice.Conclusions SRF probably interrupts the developing process of eyelid in early embryo of B6-Co mice.

Objective To evaluate the effects of soy isoflavones on bone density (BMD) in women in randomized clinical trials by meta-analysis. Method We searched the databases the Medline, Pubmed, and CNKI from January 1990 to October 2007 using the keywords, phytoestrogen, isoflavone, soy, genistein in combination with bone. We only included the studies of randomized clinical trial, in which the data of BMDs at lumbar spine, total hip or femoral neck prior to and post isoflavone intervention or their relevant changes and their standard deviation or 95% CI in women were provided. Results Sixteen papers (1304 women, 91% postmenopausal) were included and a mean daily dose of 73 mg supplemental soy isoflavones resulted in weighted mean (%)(95%CI) difference in yearly BMD changes of 18.3 (2.0%,CI 6.0~30.6) and 3.3(0.40%,CI 0.5~6.1) mg/cm2 at the lumber spine and total hip, respectively. Subgroup analyses showed that the effects were more pronounced in those with the isoflavone dose ≥80 mg/d than those of

ObjectiveTo assess gene chip application value in detecting pathogenic bacteria in intracranial infection cases.MethodsPrimers and probes aiming at the specific DNA sequences of 4 kinds of common pathogenic bacteria and 6 kinds of common drug resistance genes (DRGs) were designed and used to identify the bacteria and DRGs among 30 cerebrospinal fluid (CSF) specimens (12 positive,18negative in CSF culture) from patients with intracranial infection using multiplex polymerase chain reaction (mPCR) and gene chip.The results of gene detection were compared with those of CSF culture and drug sensitivity testing.ResultsBacteria were identified and DRGs were detected in 15 specimens; DRGs and 16S gene were detected in 8 specimens; neither bacterium nor DRG was detected in 7 specimens.ConclusionGene chip technique is characterized by its relative sensitivity and rapidity of detecting the pathogenic bacteria in CSF of intraeranial infection cases.

Objective To study the clone sequencing and expression of fibroblast growth factor 10(Fgf10) gene in corneal opaci-ty (B6-Co) mouse .Methods Normal mice mate with B6-Co mice ,the skin tissue separation from B6 and B6-Co mice at embryo 16 . 5 d ,total RNA extraction and reverse transcription ,the target gene was fragment amplification by RT-PCR ,connection with T vec-tor ,transformed to competent cells ,selection positive clone ,sequencing analysis .The gene expression in B6-Co mice was detected by real-time PCR .Results The base A inserted between 1 914 and 1 915 in Fgf10 gene by sequencing .The expression of Fgf10 was significant down regulation in B6-Co mice by real-time PCR(P< 0 .05) .Conclusion Fgf10 is relevant with phenotype of B6-Co mouse ,and the regulation mechanism was expected further study .

Objective:To study the molecular mechanism for momordin in inducing apoptosis of multidrug-resistant human chronic leukemia K562/A02 cells. Methods:The growth inhibition value of K562/A02 cells was detected by CCK-8 method. Cell apoptosis was analyzed by Annexin Ⅴ flow cytometry (FCM) and cell morphological examination. FCM was also used in determining expression of P-glycoprotein, p53 protein, bcl-2 protein and caspase activity. Results:Momordin inhibited the proliferation of K562/A02 cells in a dose-dependent manner. It also induced cell apoptosis, reduced the expression of P-glycoprotein, p53 protein and bcl-2 protein, and increased caspase-3 and caspase-8 activity.Conclusion:Momordin reversed the inhibition of apoptosis in multidrug-resistant K562/A02 cells. The molecular mechanism may be related with down-regulation of expression of p53 protein, P-glycoprotein, and bcl-2 protein and up-regulation of caspase-3 and caspase-8 activities.

Objective To explore the relationship between the distribution of infected snails and transmission of acute schistosomiasis in hilly regions.Methods The data concerning the distribution of the infected snails and acute schistosomiasis in Shitai County,Anhui Province from 1999 to 2008 were collected and analyzed.Results The sehistosome infection rate of human increased as the distance between the settings with infected snails and activity sites of humans shortened.Conclusions Acute infection of schistosome of human is associated with the distance between the settings with infected snails and activity sites of them.Strengthening the measures of snail control in key regions,protecting key populations and carrying out health education for schistosomiasis control are important approaches to control the transmission of acute schistosomiasis in hilly regions.

Background Pluripotent stem cell-derived retinal pigment epithelial (RPE) cells holds great promise for the treatment of age-related macular degeneration (AMD) and retinitis pigmentosa (RP),but the poor induction efficiency and the according high cost of RPE differentiation hindere its clinical applications.Curcumin is proved to have a promoting effect on the induced differentiation of embryonic stem cells (ESCs).However,the mechanism of curcumin on differentiation of human ESCs into RPE-like cells remains unclear.Objective This study aimed to explore the underlying molecular mechanism of curcumin on directed differentiation of human ESCs into RPE-like cells.Methods Human ESCs strains were cultured in the Matrigel-coated 6-well plate with mTeSRTM 1 medium until over-confluence,and basic fibroblast growth factor was withdrawn there after to induce automatic differentiation.Curcumin at the final concentration 1 μmol/L was added in the first day of differentiation for 24 hours,and the cells without curcumin in the medium served as the control group.Total RNA and protein were extracted at 3 weeks and 5 weeks after induction.RT-PCR,Western blot and immunofluorescence were performed to examine the expressions of the biomarks of stem cells and RPE cells as well as Wnt/β-catenin signaling pathway components.The endocytosis of polystyrene microsphere by induced RPE (iRPE) cells was investigated to verify their function of phagocytosis which features RPE cells.Results Pigmented cells were found from 3 weeks through 5 weeks after induction in the curcumin group,but only less pigmented cells were seen in the fifth week after induction in the control group.In the third and fifth week after induction,the relative expression levels of NANOG mRNA in the iRPE cells were significantly lower than those in the control group (t =13.086,P =0.022;t =34.186,P =0.004),and the relative expression levels of Pax6,RX,CRALBP and RPE65 mRNA were higher in the curcumin group than those of the control group (all at P＜0.01).Western blot assay showed that the expressing bands for CRALBP,RPE65 and MITF enhanced in iRPE cells with a similar appearance in human RPE cells.However,these expressions were all absent in human ESCs.Immunofluorescence staining showed the positive expressions of Pax6,MITF and ZO-1 in cytoplasm of iRPE cells in the curcumin group with a purified efficacy 100％.The fluorescence dye-doped polystyrene microspheres in cytoplasm were obvious in the iRPE cells like positive controls,but the polystyrene microsphere was absent in the negative controls.From 3 weeks through 5 weeks after induced,the relative expression levels of Lef1,MYC and TCF7 mRNA (the dwnstream target genes of Wnt signaling pathway),FZD3 mRNA (Wnt receptor),Wnt2B mRNA (Wnt ligand) and Wnt7B mRNA were significantly reduced in the curcumin group compared with the control group (all at P＜0.01).Conclusions Curcumin promotes the differentiation of human ESCs into RPE-like cells by stimulating the activation of Wnt signaling pathway,and therefore accelerate the differentiation and mature of iRPE cells.