Objective To investigate the effect of lipoteichoic acid from Bifidobacterium loaded with magnetic particle nano-sized-Fe3O4(nano-sized-Fe3O4-BLTA)on several cytokines in vivo expression in beterologous graft model for human gastric cancer in nude mice.and to analyze the inhibition mechanism of nano-sized-Fe3 O4-BLTA on BGC-823 human gastric carcinoma transplanted tumor.Methods Forty male nude mice(BALB/e.nu/nu)were used for in vivo transplant tumor model.the experimental animals were randomly divided into five groups to administrated by several dosages of Hano-sized-Fe3 O4-BLTA,and were executed after 12 d.The tumors were collected and photographed.and the tumor tissues were used for differ-ent assays for measuring tumor inhibition.The peritoneal fluid was extracted to isolate the macrophages for cytokines assays.Using irmnunohistochemical staining for vascular endothelial growth factor(VEGF)and CD3 l in tumor to investigate the tumor inhibition rate.Double antibody sandwich ELISA was used to detect the level of cytokine change.Results The mice treated with low dosage(10 μg/d)of nano-sized-Fe3O4-BLTA.the growth inhibifion rate of tumor was 49%,and the levels of VEGF(0.0224±0.0763)and CD31 (57.000 4±6.790)were lower than other treated groups(P<0.01).The high dosage(100μg/d)and me-dium dosage(50μg/d)of nano-sized-Fe3 O4-BLTA groups were significant difference(P<0.01)on the content of cytokines excreted by macrophages.The level of TNF-α(39.4040 ±-1.5590)induced by the low dosage group was higher relatively(P<0.01).Conclusion Nano-sized-Fe3O4-BLTA exerts an inhibiting effect on the growth of human gastric cancer in nude mice.Using nano-sited-Fe3 O4-BLTA,LTA can get more permeability and improve the therapy effect.which will be a new drug on the stomach cancer targeted therapy.

Objective A kind of quantitative C reactive protein (CRP) test kit was developed with colloidal gold lateral flow method. Method The kit was prepared with double antibody sandwich technology, and by material optimization and strict process control to improve performance. Quantitative assay was realized by a specialized lateral flow reader. The kit performance was evaluated with series of tests and clinical trial. Results The kit was developed with functional sensitivity≤1 mg/L, linear range 1-200 mg/L, CV<15%and with stability of 12 months. 220 samples clinical trial showed 98.6%of coincidence rate. Pearson Correlation coefficient r is 0.987, which showed no significant difference in performance compare with control kit. Conclusion A quantitative CRP test kit was developed with easy to operating and good stability, Which can be used for point of care testing or laboratory testing.

To study the morphological characteristics of hepatic stem cells and the expression of HGF, IGF-I, TGFbeta1 and their receptors in human embryonic livers at 3-5 weeks of gestation. The SABC immunohistochemical method with HE counterstaining was employed. We found that the hepatic bud formed at the end of the 3rd week. At the 4th week, the cells of hepatic bud migrated into the septum transversum mesenchyme and formed the hepatic cords. The hepatic cells at 3-4 weeks displayed the typical characteristics of immature cells: small size, a round or ovoid nucleus with dark color, scant cytoplasm with slight blue and a high ratio of nuclei/cytoplasm. They were positive for alpha-Fetoprotein (AFP), c-Met and negative for cytokertin 19 (CK19), and proliferating cell nuclear antigen (PCNA). At the 5th week, compared to those at the 4th week, the number of cells within the hepatic cords increased. But the cells at the 5th week were homogeneous and displayed the typical characteristic of immature cells. Those cells began to express PCNA at the 5th week. The hepatic cells at the 5th week were positive for insulin-like growth factor I (IGF-I), transforming growth factor beta1 (TGFbeta1) and their receptors, and were negative for hepatocyte growth factor (HGF), while HGF were positive in the cardiac cells and septum transversum mesenchyme. The results indicated that the cells of hepatic bud and cords were the hepatic stem cells. The difference of morphology and proteins expression at 3-5 weeks of gestation inferred that those stem cells belong to different developmental stage. AFP and c-Met were the markers of hepatic stem cells at the early stage of human embryo. HGF, IGF-I, TGFbeta1 and their receptors may involve in regulating the development of early embryonic human liver.
Embryo, Mammalian ; Gestational Age ; Hepatocyte Growth Factor ; biosynthesis ; genetics ; Humans ; Insulin-Like Growth Factor I ; biosynthesis ; genetics ; Liver ; cytology ; metabolism ; Proto-Oncogene Proteins c-met ; biosynthesis ; genetics ; Receptor, IGF Type 1 ; biosynthesis ; genetics ; Stem Cells ; cytology ; Transforming Growth Factor beta ; biosynthesis ; genetics ; Transforming Growth Factor beta1

Objective To analyze the etiological characteristics of human Brucella strains isolated, and to improve the precision in control and prevention of brucellosis. Methods In 2016, blood samples were collected from patients in Jingyuan County Gansu Province, and tested via the Rose-Bengal Plate Agglutination Test (RBPT) and the tube agglutination test methods,and serological positive blood samples were inoculated to bidirectional blood culture bottle for culturing, and further identified by traditional biological classification method and the Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis species-specific PCR (AMOS-PCR). Multiple-locus variable number tandem repeat sequence analysis (MLVA) -16 was used to detect molecular typing and do cluster analysis. Results The isolated strain was identified by the traditional biological classification method, bacteria could grow in thionine and reddened dye, A and M factors agglutination tests were positive, Bk2phage treatment of bacterial strain cracking, but Tb, Wb phages were not cracked. AMOS-PCR amplification result showed a 731 bp band, which was a strain of Brucella melitensis. The results of MLVA-16 showed that there was a difference in the number of repeats on some Variaable Number of Tandem Repeat(VNTR)sites of the isolated strain. Clustering analysis showed that, the isolated strain was clusted into the same clade with the clustering of Brucella melitensis type 3 from GS-201605 in Gansu. And the clustering was similar compared with that of Zhejiang, Guangdong, Fujian and Yunnan. Conclusion Human brucellosis is a inputting transmission in Gansu Province, there is a genetic variation of genotype 3 sheep Brucella between Gansu Province and other domestic provinces.

Loofah seeds ribosome inactivating protein luffin-α was fused with a tumor-targeting peptide NGR to create a recombinant protein, and its inhibitory activity on tumor cells and angiogenesis were assessed. luffin-α-NGR fusion gene was obtained by PCR amplification. The fusion gene was ligated with pGEX-6p-1 vector to create a recombinant plasmid pGEX-6p-1/luffin-α-NGR. The plasmid was transformed into E. coli BL21, and the target protein was isolated and purified by GST affinity chromatography. The luffin-α-NGR fusion gene with a full length of 849 bp was successfully obtained, and the optimal soluble expression of the target protein was achieved under the conditions of 16 ℃, 0.5 mmol/L IPTG after 16 h induction. SDS-PAGE and Western blotting confirmed the recombinant protein has an expected molecular weight of 56.6 kDa. Subsequently, the recombinant protein was de-tagged by precision protease digestion. The inhibitory effects of the recombinant protein on liver tumor cells HepG2 and breast cancer cells MDA-MB-231 were significantly stronger than that of luffin-α. The Transwell and CAM experiment proved that the recombinant protein luffin-α-NGR also had a significant inhibitory effect on tumor cells migration and neovascularization. The inhibitory activity on tumor cells and angiogenesis of the recombinant luffin-α-NGR protein lays a foundation for the development of subsequent recombinant tumor-targeting drugs.
Electrophoresis, Polyacrylamide Gel ; Escherichia coli/metabolism* ; Plasmids ; Recombinant Proteins/pharmacology* ; Saporins/metabolism*

Long noncoding RNAs (lncRNAs) play a critical role in the regulation of atherosclerosis. Here, we investigated the role of the lncRNA growth arrest-specific 5 (lncR-GAS5) in atherogenesis. We found that the enforced expression of lncR-GAS5 contributed to the development of atherosclerosis, which presented as increased plaque size and reduced collagen content. Moreover, impaired autophagy was observed, as shown by a decreased LC3II/LC3I protein ratio and an elevated P62 level in lncR-GAS5-overexpressing human aortic endothelial cells. By contrast, lncR-GAS5 knockdown promoted autophagy. Moreover, serine/arginine-rich splicing factor 10 (SRSF10) knockdown increased the LC3II/LC3I ratio and decreased the P62 level, thus enhancing the formation of autophagic vacuoles, autolysosomes, and autophagosomes. Mechanistically, lncR-GAS5 regulated the downstream splicing factor SRSF10 to impair autophagy in the endothelium, which was reversed by the knockdown of SRSF10. Further results revealed that overexpression of the lncR-GAS5-targeted gene miR-193-5p promoted autophagy and autophagic vacuole accumulation by repressing its direct target gene, SRSF10. Notably, miR-193-5p overexpression decreased plaque size and increased collagen content. Altogether, these findings demonstrate that lncR-GAS5 partially contributes to atherogenesis and plaque instability by impairing endothelial autophagy. In conclusion, lncR-GAS5 overexpression arrested endothelial autophagy through the miR-193-5p/SRSF10 signaling pathway. Thus, miR-193-5p/SRSF10 may serve as a novel treatment target for atherosclerosis.
Humans ; Atherosclerosis/genetics* ; Autophagy/genetics* ; Cell Cycle Proteins/metabolism* ; Endothelial Cells/metabolism* ; Endothelium/metabolism* ; MicroRNAs/metabolism* ; Repressor Proteins/metabolism* ; RNA Splicing Factors ; Serine-Arginine Splicing Factors/genetics* ; RNA, Long Noncoding/metabolism*