4.Characterization and comparison of interferon reference standards using UPLC-MS.
Lei TAO ; De-ning PEI ; Chun-mei HAN ; Wei CHEN ; Chun-ming RAO ; Jun-zhi WANG
Acta Pharmaceutica Sinica 2015;50(1):75-80
The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.
Amino Acid Sequence
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Chromatography, High Pressure Liquid
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methods
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Interferons
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standards
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Mass Spectrometry
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methods
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Molecular Weight
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Oxidation-Reduction
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Peptide Mapping
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Protein Processing, Post-Translational
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Reference Standards
5.Hemorrhage after duodenopancreatectomy
Cai-De LU ; Dan WU ; Feng QIU ; Zhi-Long YAN ; Han-Ting LING ; Ying-Chun SHENG ; Sheng-Dong WU
Chinese Journal of General Surgery 2000;0(12):-
Objective To study the sources and the relationship between the management and the outcome of hemorrhage after cephalic pancreatoduodenectomy.Methods The clinical data of 370 patients who underwent pancreatic resection at the Lihuili Hospital and the Second Affiliated Hospital of Zhejiang University were retrospectively analyzed.Results Postoperative bleeding occurred in 35 patients with 11 deaths.Among those intraabominal bleeding occurred in 14 cases and gastrointestinal hemorrhage occurred in 22,with one case suffering from both.Bleediug developing within 72 hours after operation in 12 cases (early-stage group),which was caused by improper intraoperative homeostasis.In other 23 cases,bleeding 72 hours after operation(later stage group)was caused by the erosion following pancreatic and/or bile leakage.Relaparotomy was performed in 13 cases and endoscopic homeostasis was performed in 3. Relaparotomy or endoscopic homeostasis was superior to that of conservative therapy in the early-stage group (P0.05).Pancreatic or bile leakage was identified as the significant risk factors for the postoperative bleeding.Conclusions In order to prevent the postoperative hemorrhage and to reduce the mortality of pancreatic resection,skillful techniques,expeditious homeostasis,proper management of stump pancreas and the prevention of pancreatic and bile leakage are essential.
6.Evaluation of reverse transcription loop-mediated isothermal amplification for detection of avian influenza A H5N1 virus.
Qi-Ming LI ; Xue-Jun MA ; Han-Chun GAO ; Rui ZHOU ; Zhi-Zhou KUANG ; Yun-De HOU
Chinese Journal of Virology 2008;24(3):178-184
A simple and sensitive Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) method was established to provide a new alternative for clinical diagnosis of Avian influenza A H5N1 virus. The method employed a set of six specially designed primers that recognized eight distinct sequences of the target for amplification of nucleic acid under isothermal conditions. In current study, fifty-one experimentally infected animal specimens and viral cultures that had been tested were analyzed by RT-LAMP for NA gene and HA gene, respectively. The amplification process of LAMP was monitored in real-time by the addition of SYBR Green dye. Meanwhile, the result showed high correlation between nested PCR and RT-LAMP. The specificity of the RT-LAMP assay was confirmed by restriction enzyme digestion analysis and sequencing of the amplified product. When the sensitivity of this assay was tested by serial 10-fold dilutions of RNA molecules from specimens, it was found that the RT-LAMP method achieved theoretically a sensitivity of 10 RNA molecules. Thus, we concluded that the RT-LAMP assay has potential usefulness for rapid detection of the Avian influenza A H5N1 virus.
Animals
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Birds
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Influenza A Virus, H5N1 Subtype
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genetics
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isolation & purification
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Influenza in Birds
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diagnosis
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virology
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Nucleic Acid Amplification Techniques
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methods
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Reproducibility of Results
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Reverse Transcriptase Polymerase Chain Reaction
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methods
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Sensitivity and Specificity
7.Effect of early enteral feeding on the expression of aquaporin 1 in edematous small intestinal tissues of rats after severe burn.
Xiao-zi XULI ; Jia-han WANG ; Zhi-qing LI ; Zhao-zhui YI ; De-quan XIAO
Journal of Southern Medical University 2010;30(4):727-730
OBJECTIVETo investigate the changes in the expression of aquaporin 1 (AQP-1) in edematous small intestinal tissues of rats after severe burn and the effect of early enteral feeding on its expression.
METHODSNinety normal adult Wistar rats were randomly divided into normal control group (n=6), burn model group (n=42, with 30% TBSA III degrees) and early feeding group (n=42). Dry weight method, ELSIA and immunohistochemistry were used to observe and detect the water content and expression of AQP-1 in the intestinal tissue at 1, 4, 8, 12, 24, 48, and 72 h after the burns.
RESULTSIn the burn model group, the water content in the intestinal tissue increased at 4 h after the injury, reaching the peak level at 48 h; AQP-1 expression decreased at 8 h after severe burn and reached the lowest level at 48 h. AQP-1 expression level showed a significant inverse correlation to the water content (P<0.01). Compared with the burn model group, the rats in the early feeding group showed increased AQP-l expression and lessened edema in the small intestines, also demonstrating an inverse correlation between water content and AQP-l expression (P<0.01).
CONCLUSIONIntestinal AQP-1 expression gradually decreased and edema worsened in rats early after severe burn, reaching the lowest or the peak levels 48 h after the injury with an inverse correlation between them. Early enteral feeding can increase the expression of AQP-l in the small intestine to ameliorate the intestinal edema in rats with severe burn injury.
Animals ; Aquaporin 1 ; metabolism ; Burns ; diet therapy ; metabolism ; Edema ; metabolism ; Enteral Nutrition ; Female ; Intestine, Small ; metabolism ; pathology ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Time Factors
8.Reliability and validity of Chinese version of the Addiction Severity Index among drug users in the community.
Wei LUO ; Cai Xia GUO ; De Lin HAN ; Zhi Jun LI
Biomedical and Environmental Sciences 2012;25(6):684-689
OBJECTIVETo assess the reliability and validity of a new Chinese version of the Addiction Severity Index (ASI-C) in drug users in the community.
METHODSThree hundred and eighty-one drug users in the community in Chengdu, Sichuan province were recruited. They were interviewed with a questionnaire consisting of the ASI-C revised on the basis of the previous Chinese version and 38 were interviewed for the second time at an interval of 7 days to evaluate test-retest reliability.
RESULTSCronbach's α coefficients for the internal consistency of the scale varied from 0.49 to 0.86. Test-retest correlation coefficients ranged from 0.50 to 0.93. Criterion validity was found acceptable, as compared with the Symptom Checklist 90 (SCL-90).
CONCLUSIONThe ASI-C presented acceptable reliability and validity in a sample of drug users in the community.
Adult ; Behavior, Addictive ; psychology ; China ; Data Collection ; Drug Users ; psychology ; Female ; Humans ; Male ; Middle Aged ; Reproducibility of Results ; Surveys and Questionnaires
9.Expression of aquaporin-1 in myocardium of rats in the early stage after scald and its relationship with myocardial edema.
Zhi-qing LI ; De-quan XIAO ; Jia-han WANG ; Yue-sheng HUANG
Chinese Journal of Burns 2013;29(3):245-248
OBJECTIVETo investigate the changes in aquaporin-1 (AQP-1) expression in myocardium of scalded rats in early stage of a burn injury, and to analyze its relationship with myocardial edema.
METHODSThirty-six healthy Wistar rats were divided into normal control (n = 6, without scald) and scald (n = 30) groups according to the random number table. Rats in scald group were inflicted with 30%TBSA full-thickness scald on the back, and intraperitoneally injected with Ringer's solution for antishock treatment. Myocardium tissue from left ventricle and serum specimen in rats of scald group were collected at post scald hours (PSH) 2, 8, 12, 24, and 48 (with 6 rats at each time point). Myocardial water ratio was determined by dry-wet weight method. The distribution of AQP-1 protein in myocardium was observed with immunohistochemical staining. The expression of myocardial AQP-1 mRNA was assessed with quantitative real-time PCR. The serum content of cardiac troponin-I (cTnI) was determined with ELISA. The rats in normal control group were detected with above-mentioned method. Data were processed with one way analysis of variance and LSD test. Correlation analysis was performed between AQP-1 mRNA and myocardial water ratio, AQP-1 mRNA and the serum content of cTnI in scald group at each time point.
RESULTSCompared with that in normal control group, the myocardial water ratio in scald group was markedly increased during PSH 8-48 (P values all below 0.01), and it peaked at PSH 12 [(80.79 ± 0.12)%]. In both groups, AQP-1 was mainly expressed in endothelial cells of capillaries and pericellular membrane of myocardial cells. The expression of AQP-1 in scald group was markedly increased from PSH 2 to PSH 48. The expression of myocardial AQP-1 mRNA in scald group was markedly higher from PSH 2 to PSH 48 than that in normal control group (P values all below 0.01), and it peaked at PSH 12 [(6.2 ± 0.7)%]. The serum content of cTnI in scald group was obviously higher from PSH 2 to PSH 48 than that in normal control group (P values all below 0.01), and it peaked at PSH 12 [(5.83 ± 0.51) µg/L]. There were statistically positive correlations between AQP-1 mRNA expression and myocardial water ratio (r = 0.849, P < 0.01), AQP-1 mRNA expression and the serum content of cTnI (r = 0.973, P < 0.01) in scald group.
CONCLUSIONSAQP-1 may play a key role in the development of myocardial edema in rats with scald.
Animals ; Aquaporin 1 ; metabolism ; Burns ; metabolism ; pathology ; Cardiomyopathies ; metabolism ; Disease Models, Animal ; Edema ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Wistar ; Troponin I ; blood
10.Effect of 5-aza-2'-deoxycytidine on growth and methylation of RUNX3 gene in human pancreatic cancer cell line MiaPaca2.
Xu HAN ; Zhi-Jun TAN ; Ren-de GUO ; Zhao-Jin LI ; Yu-Liang WANG
Chinese Journal of Oncology 2013;35(1):17-21
OBJECTIVETo investigate the effect of demethylating agent 5-aza-2'-deoxycytidine (5-Aza-CdR) on the growth of human pancreatic cancer cell line MiaPaca2 and the expression and methylation of tumor suppressor gene RUNX3.
METHODSHuman pancreatic cancer cell line MiaPaca2 cells were treated with different concentrations of 5-Aza-CdR. Morphological changes of MiaPaca2 cells were observed by light microscopy. The activity of cell proliferation was analyzed by MTT assay. The changes of RUNX3 mRNA expression were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Changes of RUNX3 gene methylation was detected by methylation-specific polymerase chain reaction.
RESULTSMiaPaca2 cells were treated with 2.5, 5, 10 and 20 µmo1/L 5-Aza-CdR, respectively. The inhibition rates of MiaPaca2 cells treated for 24 h were (9.17 ± 2.15)%, (10.75 ± 2.04)%, (12.57 ± 1.64)% and (18.70 ± 1.51)%, respectively. The inhibition rates were (14.94 ± 1.68)%, (18.60 ± 1.57)%, (22.84 ± 1.58)% and (33.24 ± 1.53)%, respectively, after 48 h treatment; (21.46 ± 1.60)%, (28.62 ± 1.72)%, (35.14 ± 1.64)% and (45.06 ± 1.47)%, respectively, after 72 h treatment; and (26.35 ± 1.71)%, (34.48 ± 1.69)%, (40.05 ± 1.60)% and (49.99 ± 1.61)%, respectively, after 96 h treatment. The differences between inhibition rates of each experimental and control groups (0.00 ± 0.00)% were statistically significant (P < 0.05). At the same time, the inhibition rates of different concentration groups showed significant differences (P < 0.05). At 48 h, 72 h and 96 h, the inhibition rates of each pair concentration groups showed significant differences (P < 0.05). 5-Aza- CdR inhibited the growth of MiaPaca2 cells, and the higher the concentration, the stronger the inhibition after 24 h. 5-Aza-CdR also reversed the methylation status of RUNX3 gene, and restored the expression of RUNX3 mRNA with a dose-effect relationship.
CONCLUSIONSThe methylation of RUNX3 gene is significantly related with the occurrence and development of pancreatic cancer, and abnormal methylation of RUNX3 gene may contribute to the loss of RUNX3 mRNA expression. 5-Aza-CdR may effectively cause reversion of RUNX3 methylation, and treatment with 5-Aza-CdR can reactivate the gene expression and inhibit the cell growth. This may provide a new way for diagnosis and treatment of pancreatic cancer.
Antimetabolites, Antineoplastic ; administration & dosage ; pharmacology ; Azacitidine ; administration & dosage ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; DNA Methylation ; drug effects ; Dose-Response Relationship, Drug ; Gene Expression Regulation, Neoplastic ; Humans ; Pancreatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism