Adolescent ; Basal Ganglia Diseases ; Calcinosis ; Child ; Female ; Humans ; Pedigree

Objective To explore the expression of ATP-binding cassette transporter C2(ABCG2) in adriamycin(ADM)-resistant human esophageal cancer cells.Methods The ADM-resistant human esophageal cancer cell(Eca109/ADM) was induced by gradually increasing the ADM concentration in the culture medium of human esophageal cancer cell line(Eca109) and long time screening culture.ABCG2 mRNA and protein of ADM-resistant cells was detected by RT-PCR,flow cytometry(FCM) and Western blot.Drug excretion of Eca109/ADM cells was examined by FCM.The drug resistance index to ADM was detected by MTT.Results The expression of ABCG2 in Eca109/ADM cells was higher than that in Eca109 cells.The drug excretion of Eca109/ADM cells was stronger than Eca109 cells.The Ecal09/ADM cells drug resistance index to ADM was 3.29.Conclusion The ADM-resistant cell line Eca109/ADM was established successfully as an ideal chemoresistant cell model.ABCG2 might be involved in the drug resistance of esophageal cancer cell.

OBJECTIVE Topreparealipidderivativeofgemcitabine(Gem)anditspolymericmi-celles to overcome the disadvantages of Gem.METHODS N-benzyl-3′-acetyl-gemcitabine(BAG)was synthesized.A BAG-loaded poloxamer polymeric micelle (BAG∶poloxamer 188 =10∶1 ,mol/mol)was prepared using an injection method.The micelles were characterized with a laser particle size and elec-tric charge instru ment and negatively-stained trans mission electron microscopy.Hu man breast cancer cells MCF-7 were cultured with Gem or BAG polymeric micelles of 5,10,20,30,50,70,90 μmol·L-1 for 24,48 and 72 h,respectively.The inhibitory rate of cells was measured with an MTT method.The MCF-7 cytotoxicity of BAG polymeric micelles was investigated.A pharmacodynamic study was per-formed on the mice bearing mouse hepatocellular cancer cells H22.Intravenous (iv)and oral (ig)ad-ministration was used at the dose of Ge m 40 mg·kg -1 or BAG polymeric micelles 62 mg·kg -1 .The mice were administered on the 1 st,4th and 7th day and sacrificed on the 8th day.Tumor inhibitory rates were measured.RESULTS TheBAGstructurewasidentifiedbythinlayerchromatograph,1Hand13C NMR,infrared ray chromatograph and mass spectrum.The appearance of BAG micelles was a slightly blue suspension.The micelles were spheres according to the electron microscopic observation.Their size was 62.82 nm and the zeta potential was -18.8 mV.The half inhibition concentration (IC50)of Gem and BAG polymeric micelles was 40.6 and 90.0 μmol·L-1 ,5.0 and 14.9 μmol·L-1 ,5.0 and 1 3.6 μmol·L-1 at 24,48 and 72 h,respectively according to the MTT results.According to the in vivo results,compared with the tumor model group,Gem (ig),Gem (iv)and BAG polymeric micelles (iv and ig)had significant effect on the tumor weight of H22 cell xenograft mice (P<0.01 ).As for anti-tumor efficiency,BAG polymeric micelles (ig)were better than Gem (ig)(P<0.05);BAG polymeric micelles (iv)were better than BAG polymeric micelles (ig)(P<0.05),and BAG polymeric micelles (iv)were almostequaltoGem(iv).CONCLUSION ThelipidderivativeofGemcanbeloadedinthepoloxamer 1 88 polymeric micelles.BAG polymeric micelles show in vitro MCF-7 cell inhibition and in vivo inhibition of mouse H22 xerografts;iv or ig.BAG polymeric micelles (ig)show better anti-tumor effect than Gem (ig),indicating that BAG polymeric micelles are a promising novel anti-tumor oral preparation.

Objective To investigate the effects of substrate stiffness on the proliferation of human umbilical vein endothelial cells ( HUVEC) during dengue virus infection.Methods Polyacrylamide gels were prepared for cell culture [(0±4) kPa].The proliferation of HUVEC cultured on substrates with differ-ent stiffness was determined by using 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfo-phenyl)-2H-etrazolium,inner salt ( MTS) assay.The cycle and apoptosis of HUVEC were determined by flow cytometry analysis.Dengue virus serotype 2 (DENV-2) strains were propagated and identified by con-ventional assays.The HUVEC were infected with DENV-2 strains at a MOI of 10 and cultured on traditional plastic and hydrogel substrates, respectively.The levels of nitric oxide (NO) and endothelin-1 (ET-1) were detected by nitric acid reductase assay and double antibody sandwich ELISA.Results Young′s modulus E value of the hydrogels was (3030 ±0.44) Pa.The proliferation of HUVEC and the expression of NO and ET-1 were enhanced along the increased substrate stiffness.However, no significant differences with the cell cycle and apoptosis were observed between cells cultured on different substrates.Conclusion The stiffness of substrates affected not only the proliferation of HUVEC, but also the release of cytokines during DENV-2 infection.The development of dengue fever was associated with the decreased secretion of vascular active substances as a result of blood vessel injury.The establishment of hydrogel substrates as the model of vascu-lar basement membranes might provide a new way for the in vitro investigation of the pathogenesis of DENV infection.

Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Hippocampus ; cytology ; Neurons ; cytology ; drug effects ; Propofol ; pharmacology ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats

Objective:To study the interaction of the inflammatory cytokines expression between CD4+ T cells and primary human umbilical vein endothelial cells (HUVECs) infected by dengue virus (DENV-2).Methods:PBMC was extracted from white blood cells by density gradient centrifugation,CD4+ T cells were sorted by immunomagnetic beads.The expression of CD3 and CD4 molecules on the surface of cells was detected by flow cytometry to identify the purity of CD4+ T cells.First,HUVECs were pretreated by specific-S1 P1 receptor agonist CYM-5442 for 24 h,second,infected by DENV-2 on the titer of 103 TCID50,then co-culturing with CD4+ T cells,The relative expression of NS1 partial sequence and IL-6,L-8 mRNA of HUVECs,and IL-4,IL-17,TNF-α,IFN-γ of CD4+ T cells detected by Real-time RT-PCR.IL-6 and IL-8 secreted in cultured supematant analyzed by ELISA.Results:The purity of CD4+ T cells was (98.02±0.32) ％.The expression of NS1 gradually increased to 24 h (3.03±0.26,P＜0.001),decreased after reaching the peak.The relative expression of NS1 in the group of co-cultured with CD4+ T cells was lower than other groups.After infection,the expression of IL-6 and IL-8 were up-regulated,and the expression of IL-6 and IL-8 at each time point was significantly increased after co-culturing with CD4+ T (P＜0.01).IL-6 of CYM-5442 pretreatment group,in 24 h (28.91±2.34,P＜0.05),36 h(19.36±0.1,P＜ 0.05) and 72 h(13.84±0.82,P＜0.05) was significantly decreased,the expression of IL-8 also decreased significantly.The mRNA expression of IL-4,IL-17,TNF-α and IFN-γ in CD4+ T cells was significantly increased after co-culturing with HUVECs.After the treatment with CYM-5442 group,the expression was decreased.Conclusion:DENV-2 could infect the primary HUVECs,and the expression of NS1 was inhibited after co-culturing with CD4+ T cells.CD4+ T cells can not only enhance the activation of HUVECs infected by DENV-2,but also can be activated by the infected HUVECs infected with DENV-2.

Objective To analyze the effects of dengue virus 2 ( DENV-2 ) infection on the ex-pression of IL-29 in primary human umbilical vein endothelial cells ( HUVECs) cultured on hydrogel sub-strates .Methods Primary HUVECs were isolated and cultured on hydrogel substrates .DENV-2 stains were used to infect the primary HUVECs at a multiplicity of infection( MOI) of 10.Flow cytometry analysis was performed to detect the apoptosis and infection rate of HUVECs after 48 hours of culturing .The gene chip profiling was performed to analyze mRNA expression .The expression of IL-29 at mRNA and protein levels were measured by real-time fluorescent quantitative PCR analysis and double antibody sandwich ELISA as -say, respectively.Results Compared with 96.36%of baby hamster kidney (BHK) cells that were infected with DENV-2 stains, only 4.71%primary HUVECs cultured on hydrogel substrates were infected .The pri-mary HUVECs cultured on hydrogel substrates with or without DENV-2 infection showed no significant differ-ences with the rates of cell apoptosis and infection (P>0.05).A significant difference was observed with the expression of IL-29 at mRNA level between primary HUVECs cultured on hydrogel substrates and the cells cultured in plastic bottles (P<0.05).The results of the real-time quantitative PCR analysis and ELISA as-say showed that IL-29 was highly expressed in DENV-2 infected primary HUVECs cultured on hydrogel sub-strates as compared with those in control groups (P<0.05).Conclusion The expression of IL-29 was de-tected in DENV-2 infected primary HUVECs cultured on hydrogel substrates , which was significantly differ-ent from that in DENV-2 infected primary HUVECs cultured in plastic bottles .The successful establishment of hydrogel substrates as the model of vascular basement membranes might provide a new way for the investi -gation of the pathogenesis of DENV infection .

Objective To explore the effect evaluation of choking risk assessment and intervention on the choking prevention of homeless inpatients with psychiatricc. Methods Totally 163 homeless inpatients with psychiatric disorder were selected from July 2012 to June 2016, 56 cases from July 2012 to December 2013 were assigned to the control group, another 107 cases from January 2014 to June 2016 was set as the intervention group. The control group received routine nursing safety management, on the basis of it, the intervention group received choking risk assessment through choking risk assessment scales. To take choking risk intervention on the patients of reached choking danger threshold. The incidence of choking and choking ending were compared between the two groups and the choking risk levels and scores in the intervention group before and after intervention were also compared. Results The incidence of choking in the intervention group and the control group was 0.93%(1/107)and 10.71%(6/56)respectively. Choking ending: 2 heal, 3 aspiration pneumonia, 1 death of 6 cases in the control group and 1 case was cured, no death and aspiration pneumonia occurred in the intervention group, the statistical differences were significant (χ2=9.399,18.554, P < 0.01 or 0.05). There were 28.0% (30/107) reached level Ⅰ, Ⅱ and Ⅲ choking threshold in the intervention group and rank mean of choking risk scores were 41.29. After interventions were 8.4% (9/107) and 25.50 respectively, the statistical differences were significant (χ2=10.231, P<0.05;Z=7.511, P<0.01). Conclusions The corresponding nursing intervention on the patients of choking risk can reduce and avoid choking accidents and protect the safety of homeless inpatients with psychiatric disorder effectively and improve the quality of care.

Objective:To investigate the value of magnetic resonance cholangiopancreatography(MRCP)in the diagnosis of pancreas divisum by comparing with endoscopic retrograde cholangiopancreatography(ERCP).Methods:The MRCP and ERCP images of 8 patients with pancreas divisum were retrospectively analyzed.The diagnostic accuracy and findings by MRCP were compared with those by ERCP.Results:MRCP had a diagnostic accuracy of 87.5%(7/8)based on the result of ERCP.ERCP displayed the dominant dorsal pancreatic ducts in all 8 cases and ventral pancreatic ducts in 6 cases;MRCP also displayed the dominant dorsal pancreatic ducts in all 8 cases,but the ventral pancreatic ducts only in 3 cases.Conclusion:As a non-invasive technique,MRCP has important clinical value in the diagnosis of pancreas divisum.

Objective To investigate the prevalence of primary HIV-1 genotype drug-resistance and viral subtype from 237 treatment-naive patients in China.nethods CD4+ T cell counts,plasma HIV-1 viral load and HIV-1 gene sequencing in total 237 treatment-naive patients enrolled from 20 provinces/regions were detected for the evaluation of primary HIV genotypic drug-resistance.Results The survey of 237 treatment-naive patients from muhicenter areas including Henan Province,Yunan Province,and Shanghai showed that 9 subtypes of HIV-1 strains were finally identified.Most of patients were infected before 2003.Only 3 cases had genotypic mutations associated resistance to antiretroviral drugs,with high resistance to nucleoside reverse transcriptase inhibitors(NRTIs),moderate resistance to protease inhibitors(PIs)and high resistance to non-nucleoside reverse transcriptase inhibitors(NNRTIs).respectively.The prevalence of primary genotypic drug resistance was 1.3%(3/237)in this study.Conclusions The rate of HIV-1 primary genotypic drug-resistance is still relatively low in treatment-naive HIV/AIDS patients while 9 subtypes of HIV-1 strain was diseovered.