1.Analysis on the effect factors of vitamin D level of preschoolers
International Journal of Laboratory Medicine 2016;37(6):756-757
Objective To analysis on the 25-hydroxyvitamin D[25-(OH)D] level of preschoolers in Yangzhou ,evaluating their nutrition conditions ,to discuss the influence factors .Methods The 25-(OH) D in plasma of peripheral vein was measured in the preschoolers who came to Maternal and Child Care Service Hospital of Yangzhou receiving healthy examinations in 2014 all over the year ,SPSS17 .0 software was used to analyze the data .Results The vitamin D level of preschoolers had statistical significance be-tween different age group .Season and vitamin D level of pregnant women also contributed to the statistical significance .Conclusion The vitamin D level of preschooler children shows low ,the supplement of vitamin D of pregnant women could improve the new -borns ,reduce the rate of illness .
2.Expression of metallothionein 1H in children and adolescents osteosarcoma and effect on cell proliferation
Xinfang HOU ; Shuai LI ; Chen WU ; Shuning XU ; Ke LI ; Jufeng WANG
Chinese Journal of Applied Clinical Pediatrics 2017;32(3):208-211
Objective To detect the expression levels of metallothionein1 H(MT1 H)in children and adoles-cents osteosarcoma serums,and to analyze its relationship with clinicopathological features,and to explore the effect of MT1 H on cell proliferation of osteosarcoma cells and its mechanism.Methods Enzyme -linked immuno sorbent assay (ELISA)was performed to detect the expression of MT1 H in children and adolescents osteosarcoma serums and non-neoplastic disease serums.MT1 H vector was transfected into the osteosarcoma U2OS cells.Reverse transcription -poly-merase chain reaction(RT -PCR)and Western blot were used to detect the expression of the mRNA and protein of MT1 H,respectively.Methylthiazolyldiphenyl -tetrazolium bromide(MTT)was used to detect the cell growth.Western blot was performed to detect the expression of nuclear factor(NF)-κB,and inhibitor of κB (IκB)-αprotein. Results The expressions of MT1 H in osteosarcoma serums and nonneoplastic disease serums was (0.51 ± 0.52)μg/L and (2.17 ±0.78)μg/L,respectively,with a significant difference between the 2 groups(t =-8.966, P <0.05).The expression of MT1 H in stage Ⅰ -ⅡA andⅡB -Ⅲ was (1 .98 ±0.69)μg/L and (2.45 ±0.82)μg/L,respectively,showing a gradual increase depending on clinical staging(t =-2.343,P <0.05).The expressions of MT1 H mRNA and protein were elevated in osteosarcoma U2OS cells after MT1 H vector transfection(all P <0.05). MTT assay showed that,the A value in blank control group,blank vector group,MT1 H vector group were 0.38 ±0.03, 0.36 ±0.03,0.42 ±0.03,respectively,the cell proliferation in the MT1 H vector group was significantly promoted when compared with these in the blank vector group and blank control group(F =4.213,P <0.05)from the third day.West-ern blot showed that,the relative expression of NF -κB in blank control group,blank vector group,MT1 H vector group were 0.56 ±0.05,0.53 ±0.05,0.92 ±0.07,respectively,the relative expression of IκB -αprotein were 0.64 ± 0.06,0.62 ±0.09,0.34 ±0.08,respectively,the expression of NF -κB protein was up -regulated and the expression of IκB -αprotein was down -regulated in the MT1 H vector group when compared with those in the blank vector group and blank control group(F =44.581 ,14.927,all P <0.05).Conclusions The expression of MT1 H is increased in children and adolescents osteosarcoma serums compared with that in nonneoplastic disease serums.The clinical stage is later,the expression of MT1 H is higher.MT1 H promotes cell proliferation through regulating the NF -κB pathway.
3.Regulation of swimming motility by H-NS in Vibrio parahaemolyticus
Jie WANG ; Lei LIN ; Fengjun SUN ; Xinbo DONG ; Shuning HOU ; Dongsheng ZHOU ; Zhe YIN ; Yiquan ZHANG
Military Medical Sciences 2015;(9):694-697
Objective To investigate the regulation of swimming motility by H-NS in Vibrio parahaemolyticus(VP). Methods VP was inoculated into the semi-solid swimming agar plate containing 1% Oxoid tryptone, 2% NaCl, 0.5%Difco Noble Agar, and 0.1% arabinose followed by incubation at 37℃ for 4.5 h before the diameters of bacterial lawns were measured.Total RNAs were extracted from the wild-type (WT) strains and the hns null mutant (Δhns), and the quantitative real-time( RT)-PCR( qRT-PCR) was carried out to calculate the transcriptional variation of flaA between WT andΔhns strains.The entire promoter DNA region of flaA was amplified and cloned into the lacZ fusion vector pHRP309 containing a promoterless lacZ gene. The recombinant lacZ reporter plasmid was transformed into WT and Δhns, respectively, to measure the β-galactosidase activities in cellular extracts using the β-galactosidase enzyme assay system. Results and Conclusion The phenotype results showed that swimming motility of VP was enhanced by H-NS.The qRT-PCR and LacZ fusion results indicated that the transcription of flaA was positively regulated by H-NS.Collectively, H-NS promotes the swimming motility of VP, at least partly, by activating the transcription of flaA.
4.Determination of Valepotriates and Their Degradation Products in Valeriana Jatamansi Jones by HPLC
Wenhui HOU ; Yong LIU ; Chunguo WANG ; Guolin LIU ; Qiantong LIU ; Guoping CHANG ; Jie LIU ; Shuning SHI ; Jinli SHI
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(12):2658-2663
This study was aimed to establish an HPLC method for simultaneous content determination of valtrate, acevaltrate, and their degradation products, which were baldrinal and 11-ethoxyviburtinal, in Valeriana jatamansi Jones. The separation and quantification of 4 constituents mentioned above were performed on an Agilent ZORBAX SB-C18 (4.6 mm×250 mm, 5 μm). The mobile phase consisted of water (A) - acetonitrile (B) with an optimized gradient program. The flow rate was 1 mL·min-1. The column temperature was set at 25℃. The wavelength was set at 241 nm. And the injection volume was 10μL. The results showed that among 14 different places of V. jatamansi, the 4 contents determined were different. The contents of valtrate, acevaltrate, and baldrinal in the Yunnan Baoshan Mount were the highest. And the content of 11-ethoxyviburtinal was the highest in Yunnan Dali. It was concluded that the method was with good precision, reproducibility and stability. And it was suitable for the determination of 4 valepotriates ingredients in V. jatamansi. It also provided references for the quality control and exploitation of V. jatamansi.
5.The inhibition effects of curcumin on EMT of PTX-resistant esophageal cancer cell line and its mechanism
Ke LI ; Shuning XU ; Ying LIU ; Shuai LI ; Lei QIAO ; Xingfang HOU ; Jufeng WANG
Chongqing Medicine 2017;46(34):4753-4756,4762
Objective To establish the effect of curcumin on PTX-resistant esophageal cancer cell line EC9706/PTX and to investigate the mechanism of curcumin on the epithelial stromalization (EMT) of EC9706/PTX cells.Methods EC9706/PTX cells were established by medium concentration intermittent method.The drug resistance index and cross resistance were measured by MTT assay.The inhibitory effects of different concentrations of curcumin on EC9706/PTX cell proliferation were detected.The effects of curcumin on the morphological changes,migration and invasion of EC9706/PTX cells were examined by cytostatic staining,scratching and transwell invasion assay.The effects of curcumin on the expression of E-cadherin,N-cadherin,vimentin and fibronectin in EC9706/PTX cells at mRNA and protein levels were detected by fluorescence quantitative PCR and Western blot.The effect of curcumin on the expressions of NF-κB p65 and Snail in EC9706/PTX cells were detected by Western blot.Results The drug resistance index of EC9706/PTX was 28.4,which was cross-resistant to cisplatin and doxorubicin.Curcumin could inhibit the proliferation of EC9706/PTX cells.The migration and invasion of EC9706/PTX cells were significantly decreased under the action of curcumin at 20 μmol/L concentration.Fluorescence quantitative PCR and Western blot analysis showed that the expression of Ecadherin was down-regulated and the expression of N-cadherin was up-regulated,and curcumin reversed this phenomenon.Western blot analysis showed that the expression of NF-κB p65 and Snail protein was enhanced after PTX-resistant generated in EC cell,but curcumin reversed this phenomenon.Conclusion Curcumin can inhibit the proliferation,migration and invasion of EC9706/PTX cells.The mechanism maybe that curcumin inhibits the NF-κB-Snail pathway.
6.Evolution of chemistry and selection technology for DNA-encoded library.
Peixiang MA ; Shuning ZHANG ; Qianping HUANG ; Yuang GU ; Zhi ZHOU ; Wei HOU ; Wei YI ; Hongtao XU
Acta Pharmaceutica Sinica B 2024;14(2):492-516
DNA-encoded chemical library (DEL) links the power of amplifiable genetics and the non-self-replicating chemical phenotypes, generating a diverse chemical world. In analogy with the biological world, the DEL world can evolve by using a chemical central dogma, wherein DNA replicates using the PCR reactions to amplify the genetic codes, DNA sequencing transcripts the genetic information, and DNA-compatible synthesis translates into chemical phenotypes. Importantly, DNA-compatible synthesis is the key to expanding the DEL chemical space. Besides, the evolution-driven selection system pushes the chemicals to evolve under the selective pressure, i.e., desired selection strategies. In this perspective, we summarized recent advances in expanding DEL synthetic toolbox and panning strategies, which will shed light on the drug discovery harnessing in vitro evolution of chemicals via DEL.