Objective To explore the effect on walking ability of the orthotic insole produced by the International College of Biomechanics (ICB) when it is combined with an ankle foot orthosis (AFO).Methods A total of 40 stroke survivors with hemiplegia were randomly divided into an observation group and a control group,each of 20.In addition to conventional medical treatment and rehabilitation,the control group received AFOs and walking rehabilitation training.The observation group accepted walking training with an ICB orthotic insole combined with an AFO.The experiment lasted 4 weeks.Timed up and go test (TUGT) times,10 m maximum walking speed (10 m MWS) and Berg balance scale (BBS) ratings were recorded before and after the treatment.Results Before the treatment there was no significant difference between the group averages in terms of any of the measures.After the treatment,significant improvement was observed in all of the measurements,with the improvements in the observation group significantly better than those in the control group,on average.Conclusion An ICB orthotic insole combined with an AFO results in better improvements in the walking ability of hemiplegic subjects than an AFO alone.

Objective To investigate the chemical constituents in the stems and leaves of Arachis hypogaea.Methods The isolated compounds Ⅰ—Ⅳ were obtained by the combination of silica gel,ODS-18,and Sephadex LH-20 column chromatographies.Structural elucidation was conducted by the moedrn spectral method.Results Seventeen compounds were isolated and identified from the water extraction,and there were four norsesquiterpene compounds from butanol extraction.They are arachiside A(Ⅰ),cucubalactone(Ⅱ),roseoside(Ⅲ),and citroside A(Ⅳ).Conclusion Compound Ⅰ is a new compound named arachiside A and compounds Ⅱ—Ⅳ are isolated from the plants of Arachis Linn.for the first time.

The animal models for pharmacologic assessment of drugs against premature delivery arereviewed, which include the measure of spontaneous delivery time between the first and the second pups in term pregnancy rats, the delay in the onset of labor in rats and premature delivery artificially induced by lipopolysaccharide, interleukin 1 ? and prostaglandin F 2? in mice.

OBJECTIVE:To establish the quality standard of Chushi pill. METHODS:Microscopic identification and TLC were adopted for the qualitative identification of Cortex moutan,C. dictamni,Angelica sinensis,Rubia cordifolia and Gardeniae fructus in Chushi pill;HPLC was performed to determine the contents of paeonol and baicalin. It was performed on column of Kro-masil 100-5 C18 with mobile phase of methanol-water-phosphoric acid(47∶53∶0.2,V/V/V)at the flow rate of 1.0 ml/min,the detec-tion wavelength was 280 nm,the temperature was 25 ℃ and the volume was 10 μl. RESULTS:The microscopic identification showed microscopic characteristics of C. moutan and C. dictamni,and characteristics of A. sinensis,R. cordifolia and G. fructus were identified by TLC;the linear range of paeonol was 0.106 24-2.124 8 μg(r=0.999 9)and baicalin was 0.059 04-1.180 8 μg (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 2.06%;average recoveries were respective-ly 101.56%(RSD=1.68%,n=9)and 100.16%(RSD=1.13%,n=9). CONCLUSIONS:The method is simple,accurate and re-producible,and can be used for the quantity control of Chushi pill.

Objective To evaluate the expression and relationship of miR-100 and FZD-8, one of the major compo?nents of Wnt signaling pathway, and the correlation of their expressions with lymph node metastasis in patients with breast cancer. Methods The expression of miR-100 was determined in 50 samples of human breast cancer tissues and adjacent normal tissues by in situ hybridization. The correlation of miR-100 expression with lymph node metastasis was analyzed by Mann-Whitney U test. The expression of FZD-8 was measured in 50 samples of human breast cancer tissues and adjacent normal tissues by immunohistochemistry. The correlation of the miR-100 expression with the protein expression of FZD-8 was evaluated by Pearson rank analysis. Results The expression of miR-100 was significantly lower in human breast can?cer tissues than that in adjacent normal breast tissues [2.00 (1.00, 3.00) vs. 6.00 (3.50, 8.00)]. The miR-100 expression was lower in patients with lymph node metastasis than that in patients without lymph node metastasis [1.50 (1.00, 2.75) vs. 3.00 (2.00, 4.00)]. The expression of FZD-8 was significantly higher in human breast cancer tissues than that in adjacent normal breast tissues [8.00 (6.00, 9.00) vs. 6.00 (3.75, 9.00)]. The miR-100 expression was negatively correlated with the FZD-8 pro?tein expression in human breast cancer tissues (rs=-0.592, P<0.001). Conclusion The miR-100, as an anti-metastasis-miRNA, may involve in the metastasis of breast cancer, which may be related with the regulation of the expression of FZD-8.

Objective To explore the demographic factors and the risk of the pedicle screw insertion of the narrow pedi?cles. Methods Thoracolumbar spine thin?section CT image data of 312 adults from September 2014 to September 2015 were ana?lyzed. The pedicle width,medial and lateral cortical thickness, spongy bone thickness, spongy bone thickness/cortical thickness, e angle and screw path length of each pedicle were measured. The incidence and the distribution characteristics of the narrow pedi?cle were analyzed. Anatomic parameters and age, gender and stature were compared between the narrow pedicle group and non?narrow pedicle. The risk of the pedicle screw insertion of the narrow pedicle was assessed. Results Among the 3 081 pedicles, 74 narrow pedicles were determined as their pedicles width were less than 5 mm, and the proportion of narrow pedicle was 2.40%. Among the 312 subjects, 26 subjects were found having narrow pedicles, and the proportion of individuals with narrow pedicles in the population was 8.33% (26/312). The incidences of narrow pedicle in thoracolumbar spine were T10 0.32%, T11 0.32%, T12 0.98%, L1 7.54%, L2 2.92%. The spongy bone thickness, spongy bone thickness/cortical thickness of narrow pedicle were lower than non?narrow pedicle. However, there were no significant differences of medial and lateral cortical thickness, e angle and screw path length between the narrow pedicle and non?narrow pedicle. Difference of the mean age between the two subjects groups had no statistical significance. The percentage of female in narrow pedicle subjects group was 84.6%(22/26), which was higher than that in non?narrow pedicle subjects group (49.7%, 142/286). The mean stature of the male and female of stenosis pedicle group subjects were 163.8±1.3 cm and 152.5±4.3 cm, which were shorter than those of non?narrow subjects pedicle group (169.5±5.6 cm, 160.1±6.6 cm). The percentage of the cortical bone breakthrough by the pedicle screws of narrow pedicle group was (84.6%, 27/32), which was higher than that of non?narrow pedicle group (14.7%, 33/224). Conclusion L1 is the most common segment of thoracolumbar spine that narrow pedicle exist, which is the result of reduction of the spongy bone thickness. Narrow pedicle mostly appears in short stature female. There is high risk of cortical bone breakthrough by insertion of the posterior pedicle screws in the narrow pedicle.

Objective:To analyze, at cellular level, whether the mouse B16-F1 melanoma cells with OAZI-1 overexpression could activate antigen-presenting cells and promote the phagocytotic and antigen-presenting efficiencies of mouse peritoneal macrophage and bone marrow derived DC on tumor cells. Methods:The plasmid pcDNA3. 1(+)/OAZI-1 was transfected into B16-F1 cells by Li-pofectamine2000 reagent. The positive clones with OAZI-1 overexpression ( B16/OAZI-1 ) were identified by Western blot assay and RT-PCR. Macrophages from abdominal cavity and DC from bone marrow were collected from BALB/c mouse. The B16-F1 cells transfected with the pcDNA3. 1(+) (B16/3. 1) were used as the control cells in this experiment. B16-F1 cells and macrophages were co-cultured for 4 h at a 1∶5 ratio and DC were co-cultured with B16-F1 cells at 1∶1 ratio for 4 h. And then the phagocytotic efficiencies were assayed by flow cytometry. DC were co-cultured with B16-F1 cells at 1∶1 ratio for 24 h and then the expression of mature DC surface marker molecules CD40,CD80,CD86 were determined by flow cytometry. The DC activated by the tumor cells were co-cultured with mouse spleen lymphocytes for 24 h, and then IFN-γ content in culture medium was analyzed by ELISA. Results: Phagocytotic assay showed that,compared to the control cells,the OAZI-1 overexpression in B16-F1 cells significantly enhanced the engulfment of B16-F1 cells by macrophages ( 24. 7% vs 53. 9% ) and DC ( 8. 2% vs 13. 8%) . When DC were co-incubated with OAZI-1 overexpressed B16-F1 for 24 h,the expression levels of CD40,CD80,CD86 on the DC surface,which were the molecular markers for matured DC,increased from 24. 2%,20. 8% and 16. 4% to 46. 8%,32. 5% and 36. 1% respectively. Co-culture of tumor-activated DC with the spleen lymphocytes resulted in an increased IFN-γcontent in the culture medium(32. 9 pg/ml vs 15. 1 pg/ml). Conclusion:The tumor cells with OAZI-1 overexpression can be engulfed more efficiently by macrophages and DC. And this process can induce the maturation and activation of DCs. Matured DC could induce T cell activation and then activate the anti-tumor immune response.

Abstract: Objective To construct a shuttle vector pHT315-AaCPR100A with two spore-producing-dependent promoters and the target gene AaCPR100A in Escherichia coli-Bacillus thuringiensis. Methods The forward promoter of Cry3A, named Pro-1 (+), was amplified by PCR using pSVP27A plasmid as the template, and the target gene AaCPR100A was amplified using Aedes aegypti RNA reverse conversion cDNA as the template. The plasmid pHT315 was linearized by digestion with Hind Ⅲ and Sal Ⅰ. The forward promoter and the target gene were inserted into the linearized vector pHT315 successively by in-fusion cloning according to the transcription direction. The synthesized plasmid containing the Cry3A reverse promoter sequence was used as the template, and the Pro-1 (-) reverse promoter was amplified by PCR. The intermediate vector containing the forward promoter and the target gene was linearized by EcoR I restriction enzyme, and the reverse promoter was inserted downstream of the target gene by in-fusion cloning in the direction of transcription. Results By agarose gel electrophoresis, the forward promoter, target gene AaCPR100A and reverse promoter bands were clear and of good quality, which could be used for in-fusion cloning experiments. The two spore-producing-dependent promoters and target gene fragments were connected by In-fusion cloning. The recombinant vector pHT315-AaCPR100A was verified by PCR. The forward promoter, target gene fragment and reverse promoter were successfully amplified in the recombinant vector. Nucleotide sequencing verified that the sequencing results of the bidirectional promoter sequence and the target gene sequence were basically consistent with the sequence alignment results, which met the requirements of the construction of vector elements and proved that the recombinant vector was successfully constructed. Conclusions Based on the above results, this study proves that the recombinant shuttle vector with two spore-producing-dependent promoters can be successfully constructed by in-fusion cloning technology, laying the foundation for the construction of engineered Bacillus thuringiensis expressing dsRNA of AaCPR100A.

Objective:To compare acetyltransferase MORF level in periodontal ligament stem cells( PDLSCs) derived from healthy individuals ( H-PDLSCs) with those derived from the individuals with periodontitis ( P-PDLSCs ) . And to determine the effect of MORF on the osteogenic differentiation potential of PDLSCs. Methods: Human H-PDLSCs and P-PDLSCs were cultured and cloned with limited dilution method. H-PDLSCs were stimulated by LPS, TNF-α, IL-β and the mix of the 3 inflammatory factors to imitate inflammatory environment ( IP-PDLSCs ) . Quantitative RT-PCR and Western Blot were applied to examine different expression of MORF in H-PDLSCs and P-PDLSCs. Western Blot was applied to detect expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red staining were applied to determine osteogenic differentiation potential of H-PDLSCs with MORF knock-down. Results:Quantitative RT-PCR and Western Blot showed lower expression of MORF in P-PDLSCs compared with H-PDLSCs( P<0. 05). Western Blot revealed lower expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red stai-ning indicated osteogenic differentiation potential was inhibited in H-PDLSCs with MORF knockdown(P<0. 05). Conclusion: Peri-odontitis can suppress the expression of MORF in PDLSCs and inhibite the osteogenic differentiation potential of PDLSCs.

ObjectiveTo optimize human Hexastatin gene,to express,purify protein and conduct activity experimental research,and to provide a theoretical basis for further study of Hexastatin.MethodsHuman Hexastatin gene was optimized and synthesized.It was connected to the pET28a expression vector,induced to express by isopropyl β-D-1-thiogalactopyranoside(IPTG),and optimized induction conditions.After the ultrasonication of bacterial cells and inclusion bodies,the recombinant fusion protein was purified with Ni-NTA chromatographic column,analyzed and identified by SDS-PAGE and Western Blot,and conduct activity experimental research in vitro by MTT.ResultsConstructed production was pET28a-Hexastatin expression plasmid.The human Hexastatin protein was expressed in E.coli BL21 the high level and accounted for 45.1％ of the total bacterial protein.The purification of recombinant protein purified with Ni-NTA chromatographic column reached 90％,and the concentration was 80 μg/ml.Human Hexastatin protein can restrain the growth of C6,MCF-7 and human vascular endothelial cell (HMEC) cells,and inhibition ratio reach to 72.9％±3.6％,48.8％±2.9％,52.7％±2.5％,respectively through MTT test.ConclusionThe optimized human Hexastatin protein was expressed successfully,which confirmed the inhibition to tumour cells.It is a new way for anti-angiogenesis therapy of tumour.