Objective To study the lab-scale simulated method of three-effect dynamic countercurrent extraction from Hedyotis diffusa by multi-index optimization and SPSS software.Methods Polyphenol content,flavonoids content,scavenging activity of samples on DPPH radicals,and extracting rate were considered as the optimization indexes.Orthogonal design method was used to optimize the four main factors of alcohol concentration,ratio of solid to solution,extracting temperature,and extracting times.Results The optimum condition is 70 min extraction time,solid to solution ratio of 1∶14,50% alcohol,extracting temperature 85 ℃.Conclusion The condition of large-scale industrialized production might be explored by studying the lab-scale simulated method of dynamic countercurrent extraction from H.diffusa.

Objective To investigate the immune response of mucosal immunization of new chitosan(CS) nanoparticles coating DNA vaccine. Methods The chitosan nanoparticles containing plasmid DNA encoding H. pylori lipoprotein Lpp20 gene were prepared using a complex coacervation method and then its speciality were analyzed. We then administered the naked plasmid DNA and chitosan-DNA nanoparticles to 6-week-old female BALB/c mice by intranasal or oral mucosal routes to observe the humoral and cellular immune responses. Results Naked plasmid pcDNA3.1 ( + )/Lpp20 and chitosan-pcDNA3. 1 ( + )/Lpp20 nanoparticles both induced effective immune response in mice through mucosal vaccination. Specific IgG and sIgA antibodies of chitosan-pcDNA3. 1 ( + )/Lpp20 nanoparticles groups were higher than that of naked pcDNA3.1 ( + )/Lpp20 group. The concentration of cytokines IFN-γ and IL-4 in cultural supernatant of T lymphocytes from chitosan-pcDNA3. 1 ( + )/Lpp20 nanoparticles immunized mice increased greatly than that of control groups. After stimulated by corresponding antigen, the stimulation index of intranasal or oral delivery of chitosan-pcDNA3. 1 ( + )/Lpp20 nanoparticles group was significantly higher than that of pcDNA3.1( + )/Lpp20 group, CS group and PBS control group. Moreover, systemic and mucosal immune responses in mice induced by intranasal immunization were stronger than that of oral immunization. Conclusion Chitosan nanoparticles enhanced the immune response of pcDNA3.1 ( + )/Lpp20 DNA vaccine by intranasal or oral administration in BALB/c mice. Compared to oral administration, intranasal delivery of chitosan-pcDNA3.1 ( + )/Lpp20 DNA nanoparticles could induce stronger cellular and humoral immune responses in BALB/c mice.

Objective To investigate the effects of chrysin(ChR) on the induction of differentiation and apoptosis-promoting of HepG 2 human primary hepatocacinoma cells. Methods The HepG 2 cells were cultured in vitro, and then treated with ChR and all-trans retinotic Acid (RA), respectively, the alterations of nucleocytoplasm and tubulin arrangement after Gimsa staining and Coomassie brilliant blue staining were observed. The survival rate and the inhibitory rates of HepG 2 cells were determine by trypan blue counting method and MTT assay. The Alpha-fetoprotein(AFP) secretory amounts of the cells were detected by radioimmunoassay(RIA). The activities of alkaline phosphatase(ALP) andγ-glutamyltranspeptidase(γ-GT) were assayed by enzymatic reaction kit. The synthesis of tyrosine-α-ketoglutaric acid transaminase(TAT) in cells were investigated by Diamondstone spectrophotometry. Results After treatment with ChR or RA at 1.0~100μmol/L for 48 h, the proliferation of HepG 2 cells were inhibited significantly, compared with vehicle group (P<0.05 or P<0.01), the inhibitory potency of both ChR and RA on HepG 2 cells was equivalent and indicated in dose-dependent manner. After treatment with 10μmol/L ChR or RA for 48 h, HepG 2 cells disaggregated and grew to spindle-shape, their nuclei became smaller and the number of nucleolus were fewer. Furthermore, tubulin arrangement of cells tended to be more ordered and the tubulin synthesis increased significantly. At 24~96 hours treated with 10μmol/L ChR, the activities of TAT and ALP in cells were all increased distinctly (P<0.05, P<0.01), and the secretory amounts of AFP and the specific activities ofγ-GT were decreased significantly (P<0.05, P<0.01). Conclusion Chrysin can inhibit the proliferation of HepG 2 cells and induce them to differentiate to mature cells.

To observe different expression patterns of β-catenin and its clinical significance in colorectal cancer (CRC).Methods A total of 181 cases of CRC tissues and 30 cases of normal colorectal tissue were investigated by immunohistochemistry for the expression of β-catenin.Results The expression rate of β-catenin was 56.9％ (103/181) in CRC,and higher than that in normal colorectal tissue (P ＜ 0.05).The overexpression of nuclear β-catenin was significantly correlated with histological differentiation,lymph node metastasis and Dukes' stage in CRC (P ＜ 0.05),and no relationship with other pathological parameters,such as age,gender and the depth of infiltration.The incomplete membranous expression of β-catenin was significantly correlated with histological differentiation,the depth of infiltration,lymph node metastasis and Dukes' stage in CRC (P ＜ 0.05).The high expression of nuclear β-catenin related to histological differentiation and Dukes' stage in CRC (P ＜ 0.05).In the follow-up data of 82 cases of CRC,the expression of nuclear β-catenin was associated with poor prognosis,and the 5-year survival rate was significantly lower than that of self-control groups (P ＜ 0.05).Conclusion β-catenin plays important roles in colorectal carcinogenesis.Abnormal expression of β-catenin was related to the aggressive progression of CRC and may be helpful for evaluating the prognosis of patients with CRC.β-catenin is expected to become a new target for diagnosis and treatment of CRC in future.

Objective To evaluate the clinical application and the therapeutic effect of the Fresnel press-on prism for children ocular torticollis.Design Retrospective case series.Participants 64 patients with ocular torticollis.Methods All the patients with ocular torticollis wore Fresnel press-on prism on glasses for 6-12 months to correct small angle vertical paralytic heterotropia.Main Outcome Measures Changes of abnormal head position and stereoscopic vision when naked and glasses-wearing.Results Fresnel press-on prism could provide an improvement in the correction of symptoms in children ocular torticollis.58 patients(90.6%) had improvement of abnormal head position and three-grade binocular vision.All 55 cases(100.0%)matching examination after wearing prism showed simultaneous perception,52 eases(94.5%)had fusion,and 50 cases(90.9%)had stereoscopic vision.Chi square test showed significant difference from pre-wearing prism(P

Juvenile idiopathic arthritis(JIA)is the most common rheumatology disease in childhood period with poor prognosis.The biological agents are newly developed drugs with features of clear therapeutic targets and fast effects.But its use in JIA is still limited,so this article focuses on the clinical use experience,timming and sideffects of the biological agents on JIA.

The structural protein VP6 of rotavirus form the middle layer of the triple-layered viral capsid, playing a key role in the organization of the virion. The gene of structural protein 6 of rotavirus strain TB-Chen isolated from a clinic sample was amplified using PCR from the reverse transcription product of RV genome RNA, using pET as expression vector, a recombinant plasmid pET-VP6 containing coding sequence of VP6 protein was constructed. The results showed that the VP6 was highly efficiently expressed in E. coli BL21(DE3) cells which were transformed with the recombinant plasmid pET-VP6.The expressed VP6 protein possessed 27.4% of total cells protein, with an approximately 45kDa of molecular weight, and could be recognized by guinea pig anti-SA11 antibody on Western blot. The results obtained provide important basis for further study on structure and function of the VP6 protein.

Objective:To investigate frequencies of microsatellite instability(MSI)and loss of heterozygosity(LOH)in renal ceil carcinoma(RCC),and to discuss the relationship of clinicopathological characteristics of RCC with MSI and LOH. Methods:Twelve microsatellite markers located at chromosomes 3p,9p and 14q were selected to investigate microsatellite alterations(MSI and LOH)in 31 RCC specimens and their paired metastasis specimens by polymerase chain reaction- polyacrylamide gel elect rophoresis-ethylene dibromide(PCR-PAGE-EB)staining and sequencing.Results:The frequency of MSI could reached 61.3% and that of LOH could reach 54.8%.The highest frequency of MSI was at locus of D9S168(32.3%);the highest frequency of LOH was at locus of D3S1289(21.4%).No correlation was found between MSI or LOH and the patients' age,sex,pathology type and metastastis,except that MSI was correlated with TNM stage of RCC(P

The spores suspension of Streptomyces roseosporus D-38 irritated with 20mW He-Ne laser for 20 min were incubated on G1 medium plates containing 1. 9?g/mL of streptomycin. Ten percent of mutants increased the potency of daptomycin by streptomycin-resistance method, including the mutant LC-54, which could produce daptomycin 81. 2 mg/L, which was 39% higher than that of the beginning strain by flask fermentation.

Background There is multi-dimensional order of spatial stereopsis perception in human,however,current stereopsis examination is zero-order of position disparity.A multi-dimensional space perception model is very important for the detection of stereoacuity.Objective This study was to screen the deficit of zero-order,first-order,second-order multi-dimensional spatial stereopsis perception in amblyopia and strabismus children and to explore the association of zero-order,first-order,second-order spatial perception deficit.Methods Multidimensional spacial perception was examined in 79 children aged 4-14 years in Beijing Children' s Hospital.Nineteen normal children,19 children with ametropia amblyopia,12 children with anisometropic amblyopia,18 children with strabismus and 11 children with strabismus combined amblyopia were included this study.The random-dot and line spatial stereopsis perception in zero-order,first-order and second-order were examined with a new system of multidimensional space perception screening.Written informed consent was obtained from each subject or custodian before any ocular examination associated with this study.Results Absence of zero-order,first-order,second-order random-dot channel was found in 24 children (24/79,30.4％),18 children (18/79,22.8％) and 24 children (24/79,30.4％) respectively,with an average percentage of 27.9％.Absence of zero-order,first-order,second-order line channel was examined in 37 children (37/79,46.8％),37 children (37/79,46.8％),32 children (32/79,40.5％),with an average percentage of 44.7％.In the children with a deficiency of the zero-order spatial perception,the children who still remained the first-order or/and second-order spatial perception of random-dot accounted for 41.6％ and that of lines accounted for 43.2％.In children without deficiency of zero-order random-dot or lines spatial space perception,deficiency of first-order and/or second-order spatial perception was in 37.5％ children.Various order spatial perception deficiency was seen in children suffering from amblyopia or strabismus compared with normal group(P ＜ 0.05).Conclusions There exists spatial perception deficiency in children with amblyopia or strabismus.The patients with zero-order spatial perception absence partially remain a first-order or/and second-order spatial perception;while the patients with normal zero-order spatial perception might have first-order or second-order spatial perception deficiency.The multi-dimensional space perception model has a directive role for the training of visual information process and the treatment of spatial perceptual learning in children with amblyopia or strabismus.