1.The impacts of epirubicin and liposome doxorubicin on human breast cancer cell lines and dentritic cells
Fang YU ; Ling WANG ; Qing YAO
China Oncology 2001;0(05):-
Background and purpose:Epirubicin is one of the fi rst line chemotherapy drugs in the treatment of breast cancer, and liposome doxorubicin is a new antitumor drug that has been reported to have less cardiotoxicity and myelosuppression compared to free doxorubicin. Dentritic cells (DC) play important roles in tumor immunity. Our experiment investigated the impacts of epirubicin and liposome doxorubicin on different human breast cancer cell lines and dentritic cells, and evaluated their roles in the treatment of breast cancer. Methods:Human breast cancer cell lines, Bcap37 and MDA-MB-231, along with human dentritic cells isolated and induced into maturation, were cultured with epirubicin and liposome adriamycin at different concentrations (0, 0.25, 0.5, 1.0, 2.0, 4.0, 10.0 ?g/ml), respectively. The inhibitory effects were detected by MTT method after 24, 48, 72 h. Results:Epirubicin and liposome adriamycin could inhibit the proliferation of Bcap37 cells, MDA-MB-231 cells, and human dentritic cells. Liposome adriamycin exhibited a lighter inhibition on dentritic cells than on human breast cancer cell lines (Bcap37 and MDA- MB-231) (F=22.208, P
2.Determination of Perfluorinated Compounds in Water by Electrospray Ionization Mass Spectrometry Combined with Hollow Fiber Membrane Extraction
Yao YAO ; Jiewei DENG ; Yunyun YANG ; Ling FANG ; Haiyun ZHOU ; Hongtao LIU ; Xiaowei WANG ; Tiangang LUAN
Chinese Journal of Analytical Chemistry 2015;(7):1053-1057
In this study, hollow fiber membrane extraction combined with ambient ionization mass spectrometry ( AMS) was developed for the simultaneous determination of 7 perfluorinated compounds ( PFCs) in aqueous solution, including perfluoroheptanoic acid ( PFHpA ) , perfluorooctanoic acid ( PFOA ) , perfluorooctane sulfonate acid ( PFOS ) , perfluorononanoic acid ( PFNA ) , perfluorodecanoic acid ( PFDA ) , perfluoroundecanoic acid ( PFUdA) , and perfluorododecanoic acid ( PFDoA) . PFCs were detected in negative ion mode using selective reaction monitoring ( SRM) mode. The extraction time and the pH value of extraction solution were optimized. 13 C4-PFOS and 13 C4-PFOA were used as internal standards for quantitative analysis. The method showed good linearity with correlation coefficient values ( r2 ) greater than 0. 991 for the seven target PFCs. With the exception of PFHpA, the limit of detection ( LOD) for other six PFCs was within ranges from 0. 8 to 2. 7 ng/L while the limit of quantitative (LOQ) was from 2. 7 ng/L to 8. 9 ng/L. The enrichment factor of five PFCs was more than two hundred. The developed method was applied to detect the seven PFCs in tap water and Pearl River water, and they were all not detected. The recoveries were within the ranges of 88. 5%-108. 3% and 94. 2%-116. 7% when 40 ng/L and 400 ng/L PFCs were spiked into tap water, respectively. In terms of the Pearl River water, the recoveries were within the ranges of 75. 0%-102. 6% and 81. 2%-97. 6% when 40 ng/L and 400 ng/L PFCs were spiked, respectively.
3.Effects of Honokiol on Proliferation and Apoptosis on U937 Cells
Fang XUE ; Zhiyong CHENG ; Lin YANG ; Shihui LI ; Jingyu ZHANG ; Li YAO ; Ling PAN
Journal of Sun Yat-sen University(Medical Sciences) 2009;30(4):408-412
[Objective] To investigate the anti-proliferative and apoptosis effect induced by Honokiol (HNK) on human myeloid leukemia cell line U937 cells in vitro.[Methods] After treated with different concentration of HNK,Hoechst33342 fluorescent staining was used to detect cell apoptosis;the growth inhibition ration of U937 cells and PBMCs were analyzed by MTT assay;the apoptosis ration was detected by flow cytometry;mitochondrial membrane potential was explored by rhodamine 123 stain;Caspase3/7 protein activity kit was used to test the Caspase3/7 activity;the Caspase-3 and Caspase-7 mRNA levels were detected by real-time fluorescent relative-quantification reverse transcriptional PCR (FQ-PCR).[Results] Honokiol could significantly inhibit the proliferation of U937 cells in terms of the indexes of IC50/U937 11.8 μg/mL and IC50/PBMCs 40.3 μg/mL,and the anti-proliferative effect was in a time and concentration dependent manner;Flow cytometry analysis manifested that Honokiol could induce U937cells apoptosis by Annexin V/PI double Annexin V/PI fluorescein stain;Honokiol significantly inhibited the mitochondrial membrane potential of U937 cells and enhanced the ability of Caspase3/7 and the mRNA expression levels,but not the PBMCs.[Conclusion] HNK can inhibit U937 cells proliferation and induce cells apoptosis via activating Caspase 3/7.
4.Clinical characteristics and antimicrobial resistance of Burkholderia cepa-cia causing infection in intensive care unit patients
Ling NAN ; Ding LIU ; Hao WANG ; Ping CHEN ; Maoyuan LI ; Qingning HUANG ; Qingyong FANG ; Yao CHENG
Chinese Journal of Infection Control 2015;(11):772-775
Objective To understand the specimen sources,clinical characteristics,and antimicrobial resistance of Burkholderia cepacia (B .cepacia )isolated from infected patients in intensive care unit(ICU),so as to provide reference for guiding rational use of antimicrobial agents.Methods Clinical data of patients with B .cepacia infec-tion in an ICU between 2011 and 2014 were analyzed retrospectively,antimicrobial resistance of strains was ana-lyzed.Results A total of 267 B .cepacia strains were isolated,the major specimen sources were sputum (80.15%, n=214),blood(14.23%,n =38),and urine(3.37%,n =9).Antimicrobial susceptibility testing results revealed that B .cepacia had multiple resistance,and was naturally resistant to multiple clinically used antimicrobial agents, such as ampicillin,cefazolin,ampicillin/sulbactam,nitrofurantoin,and cefuroxime,resistant rates were all 100%;resistant rates to ceftazidime and levofloxacin were 4.12% and 3.00% respectively;resistant rate to compound sulfa-methoxazole had increased tendency(χ2 =5.885,P =0.015).Conclusion Isolation of B .cepacia in ICU increased year by year,antimicrobial resistance is serious,management and targeted monitoring of prevention and control of healthcare-associated infection should be strengthened,antimicrobial agents should be chosen according to antimi-crobial susceptibility testing results.
5.Colonization and antimicrobial resistance of pathogens in nasal vestibular of health care workers in intensive care unit
Ling NAN ; Ding LIU ; Maoyuan LI ; Qingyong FANG ; Hao WANG ; Ping CHEN ; Qingning HUANG ; Yao CHENG
Chinese Journal of Infection Control 2016;15(8):608-611
Objective To understand colonization of pathogens in nasal vestibular of health care workers (HCWs) in intensive care unit (ICU),and provide evidence for strengthening the prevention and control of healthcare-associated infection (HAI)in ICU.Methods On may 2015,colonization status of pathogens in nasal vestibular of uninfected HCWs in ICU were actively screened,bacterial culture,isolation and identification were performed.The surveyed results were analyzed and compared with antimicrobial resistance of pathogens from patients at the same stage.Results A total of 96 HCWs were surveyed,43 pathogenic strains were isolated from different HCWs’na-sal vestibular,isolation rate and carriage rate were both 44.79%.The main pathogenic bacteria was Staphylococcus aureus(n=15,34.88%),followed by Enterobacter aerogenes (n =9,20.93%)and Klebsiella pneumoniae (K . pneumoniae ,n=7,16.28%).There was a high detection rate of pathogens from nasal vestibular of doctors,HCWs who smoked frequently and those who never exercised (all P <0.05).There were 1 strain of imipenem-resistant K . pneumoniae among 43 pathogenic strains.Resistance rate of 7 K .pneumoniae from HCWs to ampicillin/sulbactam, cefazolin,and furantoin were all >50.00%,resistance rates to cefotaxime and imipenem were 28.57% and 14.29%respectively;resistance rates of 11 strains of K .pneumoniae from patients to furantoin was 100.00% during the same stage,but were sensitive to other commonly used antimicrobial agents.Resistance rate of 4 strains of Esche-richia coli (E.coli)to ampicillin was 75.00%,to gentamicin,tobramycin,levofloxacin,ciprofloxacin,and com-pound sulfamethoxazole were all 50.00%,6 strains of E.coli isolated from patients during the same period were found to be resistant to most commonly used antimicrobial agents.Conclusion Colonization rate of pathogens is high in nasal vestibular of HCWs in ICU,active screening and monitoring on colonization of pathogens in HCWs’ nasal vestibular is significant for preventing the occurrence and cross transmission of HAI among HCWs and pa-tients.
6.Cloning, Expression and Property Analysis of Arabinosidase in Pichia pastoris
Dong-Sheng YAO ; Hui-Mei TAN ; Hui HUANG ; Da-Ling LIU ; Chun-Fang XIE ;
China Biotechnology 2006;0(04):-
SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.
7.Inducement, Purification and Characterization of?-mannanase from Armillariella tabescens EJLY2098
Dong-Sheng YAO ; Xiao-Kui HUANG ; Da-Ling LIU ; Chun-Fang XIE ; Rong HU ;
China Biotechnology 2006;0(07):-
Armillariella tabescens EJLY2098 was capable of secreting p-mannanase by konjac inducement. A 34 orthogonal design was applied to determine the optimum medium of inducing mannanase by Armillariella tabescens EJLY2098. The results suggested that Armillariella tabescens EJLY2098 secreted the high-activity enzyme in the optimum medium, which was composed of 2% konjac, 1% peptone, 25% potato juice,0.3% KH2PO4,15% MgSO4?7H2O, 0.01% VitB1. Purified by DEAE-anion exchange chromatography, two eluting peaks (P1 and P2) with the p-mannanase activity were obtained, and one of them (named?-mannanase P2) was a single band by the SDS-PAGE, and the molecular weight of?-mannanase P2 was 78. 9kDa. The isoelectric point of?-mannanase P2 was estimated to be 4.0-4. 1. The optimum activity for the enzyme was found at 60℃and pH4. 0 - 6. 0, and the enzyme was stable between pH4. 5 - 6. 0. The activity of?-mannanase P2 were enhanced by Na+ and Ba2+ . This?-mannanase can be used in feed industy. a new fungi secreting?-mannanase was obtained, providing an important base for cloning mannanase gene and constructing recombin microbe expressing?-mannanase .
8.Role of Overexpressed Proinflammatory Cytokines in the Pathogenesis of Cerebral Palsy
Yang SUN ; Fang WEN ; Han-yun YAO ; Yan GUO ; Ling XIAO ; Zili CHENG ; Juncheng WAN
Chinese Journal of Rehabilitation Theory and Practice 2006;12(2):93-94
ObjectiveTo investigate the role of overexpressed proinflammatory cytokines in the pathogenesis of cerebral palsy (CP).MethodsLevels of tumor necrotic factor-α (TNF-α) and interleukin-6 (IL-6) in serum of 31 CP children, 20 healthy children (as controls), 37 neonates with CP risk factors such as hypoxic-ischemic injury and/or perinatal infection, and 20 healthy neonates (as controls) were measured by enzyme-linked immunosorbent assays (ELISA) retrospectively.ResultsLevels of TNF-α and IL-6 of CP children and neonates with CP risk factors were significantly higher than that of healthy controls (P<0.05). TNF-α level of CP children was significantly higher than that of neonates with CP risk factors (P<0.05), but there was no significant difference in IL-6 level between two groups.ConclusionOverexpressed proinflammatory cytokines play an important role in the pathogenesis of CP and may be an independent risk factor of CP.
9.The role of arecoline on hepatic insulin resistance in type 2 diabetes rats.
Hong-Yan LING ; Qi-Xin YAO ; Zhu-Qing QI ; Si-Si YANG ; Jian-Qin HE ; Kai-Fang ZHANG ; Bi HU
Chinese Journal of Applied Physiology 2014;30(3):208-212
OBJECTIVETo explore the effects of arecoline on hepatic insulin resistance in type 2 diabetes rats and to elucidate its possible mechanism.
METHODSForty five Wistar rats were fed with high fructose diet for 12 weeks to induce type 2 diabetic rat model. rats were randomly divided into 5 groups (n = 8): control group, model group and model group were treated with different dose (0, 0.5, 1, 5 mg/kg) of arecoline. After 4 weeks, the fasting blood glucose, blood lipid and insulin level measured , mRNA expression of liver constitutive androstane receptor (CAR), pregnane X receptor (PXR), glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected by reverse transcription polymerase chain reaction (RT-PCR), the protein expression of p-AKT and glucose transporter4 (GLUT4) were detected by Western blot.
RESULTS1.5 mg/kg arecoline could significantly decrease the level of fasting blood glucose, blood lipid, blood insulin level and liver G6Pase, PEPCK, IL-6, TNF-alpha mRNA level in type 2 diabetes rats. 1.5 mg/kg arecoline also could significantly increase CAR, PXR mRNA level and p-AKT and GLUT4 protein expression.
CONCLUSIONArecoline improved hepatic insulin resistance in type 2 diabetes rats by increasing the mRNA levels of CAR and PXR leading to the creased glucose metabolism and inflammation related genes expression.
Animals ; Arecoline ; pharmacology ; Diabetes Mellitus, Experimental ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; Glucose Transporter Type 4 ; metabolism ; Glucose-6-Phosphatase ; metabolism ; Insulin Resistance ; Interleukin-6 ; metabolism ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Phosphoenolpyruvate Carboxykinase (GTP) ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Wistar ; Receptors, Cytoplasmic and Nuclear ; metabolism ; Receptors, Steroid ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism
10.The correlation analysis of the physical and mental status of organ transplanted patients' family members during the patients' rehabilitation period and the long term survival of the transplanted organ
Guirong YE ; Lin YAO ; Yulan ZHANG ; Meini FANG ; Wanzhen HU ; Ling BA ; Lisha SHI ; Fen LEI ; Hongling LEI
Chinese Journal of Practical Nursing 2008;24(31):51-53
ObjectiveTo know about the correlation analysis of the physical and mental status of organ transplanted patients' family members during the patients' rehabilitation period and the long term survival of the transplanted organs.MethodsA total of 600 organ transplanted patients' family members were investigated by Zung self-rating anxiety scale (SAS) and simple coping style questionnaire.The results underwent subsequent analysis.ResultsAnxious state of organ transplanted patients' family members was more severe than that of domestic norm (P<0.01).There were some differences in terms of anxious state of family members with different genders,education backgrounds,income and the resource of medical expenditure (P<0.05).Family members most took positive coping styles,whereas fewer adopted negative coping styles (P<0.01).Positive coping styles were negatively correlated with the anxiety of family members (P<0.01) and positively correlated with long term survival of transplanted organ(P<0.05).However,negative coping styles of family members were significantly positively correlated with their anxiety (P<0.05) and were negatively correlated with long term survival of transplanted organ.ConclusionsThe anxiety generally exists in organ transplanted patients' family members.There are differences in terms of anxious state among family members of different genders,different education backgrounds,income or with the resource of medical expenditure.The more they adopt positive coping styles,the lower anxiety level they show and the longer the transplanted organ survive.Conversely,the more they adopt negative coping styles,the higher anxious level they show and the shorter the transplanted organ survive.