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Objective To establish a UPLC-MS/MS method for the determination of dexmedetomidine in human plasma and investigate the effect of obstructive jaundice on pharmacokinetics of dexmedetomidine in vivo. Methods Samples were obtained by liquid-liquid extraction. Agilent Eclipse Plus C18 column was used for chromatograph with methanol and 0.1% formic acid-water solution as mobile phase. Flow rate was 0.2 ml/min. The column temperature was 35 ℃, and the MS detection was selected in MRM mode. Results The calibration curves of dexmedetomidine showed good linearity in the ranges of 0.01−10.00 ng/ml. The results of intra and inter-day precisions were both within 15%. The recovery rate was 85.5%−93.1%. Matrix effect was 91.2%−95.6%. Samples remained stable during analysis. Compared with the control group, cmax、AUC(0−t)、AUC(0−∞) and Vz of dexmedetomidine in the patients with obstructive jaundice were increased by 63.4%, 78.9, 66.4%, 82.5%, respectively (P<0.01). CLz was decreased by 42.1%. Conclusion This method is accurate, sensitive and reproducible. It is suitable for dexmedetomidine assay in human plasma. The elimination rate of dexmedetomidine is slower in obstructive jaundice.
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Objective To evaluate the influence of atopy on the prognosis of juvenile-onset systemic lupus erythematosus (JSLE).Methods The study was performed on 60 cases with JSLE diagnosed at the Department of Pediatrics,Renji Hospital Affiliated to School of Medicine of Shanghai Jiaotong University from October 2005 to April 2015.These patients were enrolled by mixed cohort study and subdivided into atopic group(26 cases) or non-atopic group(34 cases).The clinical and laboratory data of the disease onset,disease assessment scores,medications during follow-ups and remission/flare of the disease were recorded and analyzed to compare the difference between 2 groups.Results (1) The systemic lupus erythematosus disease activity index (SLEDAI) score [(17.080 ± 5.628) scores vs (12.590 ± 4.856) scores],anti-double-stranded DNA (anti-dsDNA) [(62.590 ± 43.602) IU/mL vs (40.230 ±30.189) IU/mL],erythrocyte sedimentation rate (ESR) [(59.150 ± 40.315) mm/1 h vs (40,350 ± 31.865)mm/1 h] were significantly elevated at onset in the atopic group compared with non-atopic controls (all P ＜ 0.05),while the complement C3[(0.450 ±0.218) g/L vs(0.640 ±0.333) g/L],C4 [(0.047 ±0.024) g/L vs(0.116 ±0.172) g/L] in atopic group was lower than those in the non-atopic group (all P ＜ 0.05).(2)During the follow ups of 1 and 6 months to 1 year,the JSLE patients with atopy always had higher SLEDA1 score compared with the non atopic controls(all P ＜ 0.05).(3)For medications,the daily cumulative glucocorticoid dose received by patients in the atopic group were larger than that of the non-atopic group,and the number of immunosuppressive agents used in the atopic group was more than that in the non-atopic controls (P ＜ 0.05).(4) During the 1-year follow-up,the rate of disease relapse in the atopic group was higher than that in the non-atopic group and the atopic group also needed much more time to reach disease remission (P ＜ 0.05).Conclusion JSLE patients combined with atopy may have an adverse influence on the prognosis of JSLE.
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Systemic lupus erythematosus is a kind of autoimmtne disease which may lead to multisystem involvement,with high mortality.As is well known,the prognosis of children is worse than that of adults.Many factors such as genetic factor,environmental factor and the self factors affect the survival of children with systemic lupus erythematosus.In order to improve the diagnosis,treatment and prognosis,this article reviews the recent research of the main factors affecting the prognosis of children with systemic lupus erythematosus.
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Objective To evaluate the expression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT-1) in ovarian cancer cell lines, and to investigate the biological effects of down-regulated MALAT-1 on OVCAR3 cells.Methods qRT-PCR analysis was used to examine the expression level of MALAT-1 gene in ovarian cancer cells, including ES-2, A2780, SKOV3 and OVCAR3 cell lines.For functional research, four shRNA oligos specially targeting MALAT-1 and a empty vector were designed and constructed into pGPU6/GFP/Neo, then transfected into OVCAR3 cells.qRT-PCR was used to confirm the effective suppression of MALAT-1.Changes of proliferation and adhesion of cells were analyzed by CCK-8 and adhesion assays.Wound-healing, transwell migration and invasion assays were used to examine migration and invasion of MALAT-l-silencing cells in vitro.Results The expression of MALAT-1 gene in OVCAR3 cells was high, and qRT-PCR results confirmed successfully the knockdown of MALAT-1 after transient transfection.After successful suppression of MALAT-1, the proliferation, wound-healing and adhesion ability in vitro were inhibited to some degree.In transwell migration assay, the number of migration cells in MALAT-1-silencing group was 52.17±4.48, which is much less than that in the negative and control groups (286.50± 12.23 and 295.67±6.96, respectively).In invasion assay, the number of invasion cells passing the transwell membrane in MALAT-1-silencing group (37.33±2.40) was also decreased significantly, compared to that in the negative and control groups (239.00±15.72 and 222.67±20.85, P ＜ 0.05).Conclusions shRNA-mediated silence of MALAT-1 can effectively inhibit the proliferation, adhesion, migration and invasion abilities of ovarian cancer cell line OVCAR3 in vitro, indicating MALAT-1 is expected to be a target gene for the treatment of ovarian cancer.
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<p><b>OBJECTIVE</b>To investigate the reversal effects of different concentrations of DNA methylation inhibitor, 5-aza-2-deoxycytidine, on the hypermethylation of maternally expressed gene 3 (MEG3) gene promoter, and then the inhibitory effect of restoration of MEG3 expression on the proliferation of ovarian cancer cells.</p><p><b>METHODS</b>Human ovarian cancer OVCAR3 cells were treated with various concentration of 5-aza-2-deoxycytidine (0, 1, 5, 10, 20 µmol/L, respectively) for 6 days. Then the methylation status of MEG3 promoter was detected by methylation specific PCR (MSP). The alteration of MEG3 gene expression was detected by RT-PCR. Cell proliferation was determined by MTT assay and EdU incorporation assay.</p><p><b>RESULTS</b>After treated with 5-aza-2-deoxycytidine, the methylation status of MEG3 in the 0, 1, 5, 10, 20 µmol/L 5-aza-2-deoxycytidine groups were 1.00 ± 0.00, 0.79 ± 0.00, 0.67 ± 0.00, 0.65 ± 0.03 and 0.61 ± 0.01 folds, respectively (P < 0.05 for all). The relative expressions of MEG3 mRNA in the 0, 1, 5, 10, 20 µmol/L 5-aza-2-deoxycytidine groups were 1.00 ± 0.00, 2.04 ± 0.16, 2.44 ± 0.17, 3.19 ± 0.34 and 5.34 ± 0.39, respectively (P < 0.05 for all). In contrast to the negative control, the inhibition rates of the OVCAR3 cell growth were increased significantly when treated with 1, 5, 10, 20 µmol/L 5-aza-2-deoxycytidine in 2, 4 and 6 days. There were (40.78 ± 0.80)%, (35.65 ± 0.33)%, (31.81 ± 0.66)%, (27.33 ± 1.27)% and (17.75 ± 1.85)% of EdU-positive cells in the 0, 1, 5, 10 and 20 µmol/L 5-aza-2-deoxycytidine groups (P < 0.01 for all).</p><p><b>CONCLUSIONS</b>Maternally expressed gene 3 promoter hypermethylation is reversed by 5-aza-2-deoxycytidine in ovarian cancer cells. The downregulation of MEG3 gene might be resulted from the methylation, and the re-expression of MEG3 partly contribute to the growth inhibition of epithelial ovarian cancer cells.</p>
Subject(s)
Female , Humans , Antimetabolites, Antineoplastic , Pharmacology , Azacitidine , Pharmacology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , DNA Methylation , Neoplasms, Glandular and Epithelial , Ovarian Neoplasms , Promoter Regions, Genetic , RNA, MessengerABSTRACT
Objective To study the relationship between the methylation status of CPG islands in MEG3 gene promoter region of epithelial ovarian cancer and its clinical and pathological features. Methods The promoter methylation status was evaluated by MSP (methylation-specific polymerase chain reaction ) in 47 cases of ovarian cancer tissue and 15 cases of normal control. Results The methylation ratio (42.6%) of the MEG3 genes in the ovarian cancer was statistically significantly higher (P = 0.035 ) than that (13.3%) in the normal control. The methyation rate of the group with an age > 60 years old was slightly higher than that of the group with an age≤60 years old, without statistically significant (P > 0.05), so was observed in ovarian cancers of stage Ⅰ andⅡ than that in stage Ⅲ and Ⅳ. There were also no significant differences in MEG3 gene methylation positive rate neither in different pathological grading nor in various ovarian cancer tissues (P > 0.05). Conclusion Abnormal methylation in MEG3 gene may be associated with epithelial ovarian cancer , but no relation to its clinical pathology.
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<p><b>OBJECTIVE</b>To provide theoretical evidences for the identification and selective breeding of different major varieties of Rehmannia glutinosa by comparing their leaf characters such as the shapes, non-glandular hairs and the size and density of stomata.</p><p><b>METHOD</b>The length, width and the ratio of the length to the width of leaves were measured, and the density and length of the non-glandular hair, the density, size and density of the stomata in the epidermis were measured by scanning electron microscope.</p><p><b>RESULT</b>The results showed that there were obvious differences in length, width and the ratio of the length to the width of leaves, the density and length of the non-glandular hair, the density and size of the stomata in the epidermis.</p><p><b>CONCLUSION</b>The morphological characters of the leaves mentioned above may serve as the identification standards of major varieties of R. glutinosa.</p>
Subject(s)
Plant Leaves , RehmanniaABSTRACT
Objective Survey the classification on diseases of patients in hospital. Discuss personnel arrange-ment of nursing staff. Methods Survey and star sickbed number,CD rate/month,nurse number accounted on nursing level and sickbed-nurse ratio in 2007, discuss personnel arrangement of nursing staff. Results It is different that the nurse number accounted by two means, Z=2.234,P=0.025. The correlation about CD rate and nurse number in theories: r=0.782,p=0.004, nurse number in theories= CD ratex0.51-17.11, F=16.543,p=0.003.Conclusion CD rate should be reasonable personnel arrangement of nursing staff.