Objective To investigate the influence of lentivirus mediated short hairpin RNA(shRNA)target to human papilloma virus(HPV)16 E6 on invasive ability of cervical cancer Caski cells.Methods Lentivirus was produced after shRNA target to human papilloma virus(HPV)16 E6 and to nonsense was cloned to lentivirus work vector.Infection ratio was assessed by assay of EGFP positive cells of Caski.Total mRNA of E6 was determined by RT-PCR after Caski cells were infected by lentivirus.The change of E6 protein expression was analyzed by Western blot.The invasive ability of Caski cells was assayed employing Transwell.Results The optimal MOI (Multiplicity of infection)of lentivirus to Caski was 2.5.Total mRNA and protein of E6 were decreased (by 70%and 63%)in interfering group compared with control group.The invasive ability of Caski cells also reduced after infected by lentivirus.Conclusion shRNA mediated by lentivirus can inhibit expression of HPV16 E6 and invasive ability of cervical cancer cells.

Hematologic Diseases ; Hemolysis ; Humans ; Infant, Newborn

Adolescent ; Adult ; Amino Acid Sequence ; Genotype ; Homozygote ; Humans ; Hypoprothrombinemias ; etiology ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; Pedigree ; Phenotype ; Young Adult

Objective The population structure and ecological distribution of endophytic fungi in Changium smyrnioides from the different habitats and growing phases,and the effects of fungal elicitor on the cell biomass and polysaccharide accumulation were studied in this paper.Methods The isolation,culture,and identification of microorganism,and plant cell suspension culture technology were adopted;And relative data were analyzed by the statistical methods.Results In four producing areas,116 strains were isolated and classified into eight genera.The dominant populations were Fusarium LK.ex FR.,Geotrichum LK.,and Alternaria Nees.The population structure of endophytic fungi obviously changed at the different growing phases.Species and quantity of endophytic fungi were plentiful at the seedling stage and bud stage,and especially at the bud stage the isolation rate and isolation frequency were more than 30% and 19%,respectively.Some endophytic fungi had the obvious area and tissue specificity.Compared with the control by adding the elicitor of Fusarium sp.3,the yields of cell biomass and polysaccharide were increased to 31.86% and 38.01%,respectively.Conclusion Endophytic fungi in C.smyrnioides have abundant biodiversity.And there is close relationship between the population structure and distribution of endophytic fungi with ecological conditions.And fungal elicitors could obviously enhance the yields of cell biomass and polysaccharide of C.smyrnioides.

OBJECTIVE: To determine the chlorpyrifos in human blood by liquid chromatography-tandem mass spectrometry and to validate its application in poisoning cases. METHODS: The samples were extracted by a simple one-step protein precipitation procedure. Chromatography was performed on a Capcell Pack C18 MGII column (250 mm x 2.0 mm, 5 μm) using an isocratic elution of solvent A (0.1% formic acid-water with 2 mmol/L ammonium acetate) and solvent B (methanol with 2 mmol/L ammonium acetate) at 5:95 V:V). RESULTS: The linear ranged from 5 to 500 ng/mL (r = 0.998 7). The limit of detection (LOD) and the lower limit of quantification (LLOQ) were 2 ng/mL and 4 ng/mL, respectively. For this method, the precision and accuracy of intra-day and inter-day were < 10% and 97.44%-101.10%, respectively. The results in stability test of long-term frozen were satisfied. The matrix effect, recovery and process efficiency were 64.97%-86.81%, 76.70%-85.52%, and 55.57%-66.58%, respectively. CONCLUSION This method can provide a rapid approach to chlorpyrifos extraction and determination in toxicological analysis of forensic and clinical treatment.
Chlorpyrifos/blood* ; Chromatography, High Pressure Liquid/methods* ; Chromatography, Liquid/methods* ; Humans ; Limit of Detection ; Poisoning ; Reproducibility of Results ; Tandem Mass Spectrometry/methods*

Objective To investigate the incidence and influencing factors of healthcare-associated infection (HAI) in inpatients with nasopharyngeal carcinoma during radiotherapy period.Methods The occurrence of HAI among patients with nasopharyngeal carcinoma in a tertiary first-class cancer hospital in Jiangsu Province between July 2012 and June 2014 were analyzed retrospectively.Results A total of 1 396 patients were investigated,the incidence of HAI was 2.29%,case incidence of HAI was 2.44%.The most common infection site was oral mucosa (n =24, 70.59%),and most infection occurred 2-4 weeks after the start of the radiotherapy.A total of 38 pathogenic iso-lates were isolated,24 (63.16%)were gram-positive bacteria,12 (31 .58%)were gram-negative bacteria,and 2 (5.26%)were fungi.Incidences of HAI were high in patients >50 years old,with chemotherapy,length of hospital stay>60 days,and used at least 2 kinds of antimicrobial agents (all P <0.05).Conclusion Prevention and control of HAI in patients with nasopharyngeal carcinoma during radiotherapy period should be strengthened,especially for the elderly,patients with chemotherapy,long length of hospital stay,and extensive use of antimicrobial agents.

Objective To determine the chlorpyrifos in human blood by liquid chromatography-tandemmass spectrometry and to validate its application in poisoning cases. Methods The samples were extracted by a simple one-step protein precipitation procedure. Chromatography was performed on a Capcell Pack C18 mG II column (250 mm×2.0 mm, 5μm) using an isocratic elution of solvent A (0.1% formic acid-water with 2 mmol/L ammoniumacetate) and solvent B (methanol with 2 mmol/L ammoniumacetate) at 5∶95 (V∶V).Results The linearranged from5 to 500ng/mL (r=0.9987).Thelimitofdetection (LOD) and the lower limit of quantification (LLOQ ) were 2 ng/mL and 4 ng/mL , respectively. For this method, the precision and accuracy of intra-day and inter-day were <10% and 97.44%-101.10%, respectively. The re-sults in stability test of long-termfrozen were satisfied. The matrix effect, recovery and process efficien-cy were 64.97%-86.81%, 76.70%-85.52%, and 55.57%-66.58%, respectively. Conclusion This method can provide a rapid approach to chlorpyrifos extraction and determination in toxicological analysis of forensic and clinical treatment.

Objective To investigate the expression and correlation of hypoxia-inducible factor-1?(HIF-1?),vascular endothelial growth factor(VEGF)and sFlt-1 in the preeclampsia placenta,and discuss their significance in the pathogenesis of preeclampsia.Methods Placentas were collected from 20 pregnant women with preeclampsia as study group and 15 normal pregnant women as control group.The expressions of HIF-1?,VEGF and sFlt-1 protein were semi-quantitatively analyzed with immunohistochemical assay and mRNA level was determined using reverse transcription polymerasc chain reaction(RT-PCR)technique. Results(1)the expression of HIF-1?and sFlt-1 protein in preeclampsia group obviously increased.Strong (+++)positive expression was observed in 9 and 11 cases respectively,significantly higher than in control group(2 and 3 cases)(P＜0.05),however,VEGF expression obviously reduced in preeclampsia group(P＜0.01).(2)the level of HIF-1?and sFlt-1 mRNA in preeclamptic placenta was 0.604?0.013, 0.898?0.041,significantly higher than 0.208?0.007 and 0.559?0.244 in normal placenta(P＜0.05). Although the level of VEGF mRNA increased in preeclampsia placenta,it was not significantly different from that in normal placenta(P＞0.05).The ratio of VEGF mRNA/sFlt-1 mRNA obviously reduced in preeclampsia group and was significantly lower than in control group(P＜0.05).(3)in preeclampsia group,HIF-1?mRNA expression was positively correlated with the expression of sFlt-1 mRNA(r=0.577, P＜0.05),and negatively correlated with the ratio of VEGF mRNA/sFlt-1 mRNA(r=-0.376,P＜0.05).Conclusion Abnormal high HIF-1?expression in preeclampsia placenta indicates that HIF-1?might play an important role in the pathogenesis of preeclampsia,possibly through affecting the cytotrophoblastic invasion and placental vascular reconstruction via the modulation of VEGF and sFlt-1 gene transcription.

Humans ; Infant ; Lead Poisoning, Nervous System, Childhood ; diagnosis ; therapy ; Male

Animals ; Carcinogenesis ; Cell Cycle Proteins ; genetics ; metabolism ; Cyclin E ; metabolism ; F-Box Proteins ; genetics ; metabolism ; F-Box-WD Repeat-Containing Protein 7 ; Gene Silencing ; Genes, Tumor Suppressor ; Humans ; Mutation ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Neoplasms ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins c-myc ; metabolism ; Receptors, Notch ; metabolism ; Signal Transduction ; TOR Serine-Threonine Kinases ; metabolism ; Ubiquitin-Protein Ligases ; genetics ; metabolism