1.Method performance verification of the detection of interleukin-6 by using IMMUNITE1000 chemiluminescence detection system
Fei SUN ; Tianpen CUI ; Lan ZHAO ; Jiayi CHEN ; Yanyan WANG
International Journal of Laboratory Medicine 2015;(13):1858-1860
Objective To verify the performance of quantitative detection of interleukin-6 by using IMMUNITE1000 chemilumi-nescence analyzer.Methods According to the requirements of International Organization for Standardization(ISO)1 5 189,serum specimen were collected and levels of IL-6 were detected.The precision,accuracy,analytical measurement range,reportable range amd normal reference range of quantitative detection of interleukin-6 by using IMMUNITE1000 chemiluminescence analyzer were verified,and its performance was evaluated.Results The coefficient variation(CV)of between-day precision of high and low value was 6.42% and 1.97% respectively,and that of within-run precision was 3.40% and 3.82% respectively.Compared the test re-sults with the target values,the bias % was 0.91%.The regression equation:Y =0.986X - 7.1 (r 2 = 0.999,P < 0.05 ).With 27 times diluted,the recovery rate was from 97% to 100%,and the clinical reportable range was 2 to 27 000 pg/mL.The 95% refer-ence interval ranged from 0 to 5.3 pg/mL.Conclusion The performance of this system meets the manufacturer′s declaration,and could satisfy the quality requirements of clinical laboratory.
2. HPTLC fingerprint analysis of Aconitum herbals in Xinjiang
Chinese Pharmaceutical Journal 2013;48(7):512-517
OBJECTIVE: To establish the HPTLC fingerprint of Aconitum plants in Xinjiang and to carry out similarity comparison, cluster analysis and principal component analysis. METHODS: Using high performance thin layer silica gel G plate, n-hexane-ethyl acetate-methanol (6.4:3.6:1) as developer, diluted bismuth potassium iodide solution as colour-developing agent, of the HPTLC fingerprints were obtained. The common pattern was explored by chemical fingerprint analysis system software. Similarity analysis, cluster analysis and principal component analysis were carried out. RESULTS: The conditions for thin layer chromatography were optimized, the spots were clear, and the separation was good; similarity analysis showed that Xinjiang Aconitum had some differences in chemical composition between different plant species, which can be broadly divided into three categories. Aconitum leucostomum Wo-rosch. and Aconitum apetalum (Huth) B. Fedtsch. had high similarity with Aconitum carmichaelii Debx. and Aconitum kusnezojfii Reichb. CONCLUSION: The established thin-layer chromatographic fingerprints have laid the methodological foundation for the study of the fingerprint of Xinjiang Aconitum plants. Aconitum leucostomum Worosch, which is a widely distributed and abundantly reserved Aconitum species in Xinjiang, has potential medicinal value and similarity with Aconitum carmichaelii Debx. and Aconitum kus-nezoffii Reichb. Which have national quality standard, and is worthy of further studies.
3.Chondrocyte differentiation of dog bone marrow mesenchymal stem cells co-cultured with Bio-gide collagen membrane in vitro
Hui XIE ; Fei YANG ; Dewei ZHAO ; Benjie WANG ; Daping CUI ; Wei WANG ; Shibo HUANG
Chinese Journal of Tissue Engineering Research 2013;(29):5282-5289
BACKGROUND:Seed cells and scaffold are two key factors for cartilage defects after osteonecrosis of femoral head using tissue-engineered method. OBJECTIVE:To explore the feasibility of Bio-gide col agen membrane combined with dog bone marrow mesenchymal stem cells into chondrocytes. METHODS:Bone marrow mesenchymal stem cells were isolated from beagle dogs by whole bone marrow blood centrifugation method and adherence screening method in vitro and cultured. Morphological changes in cells were observed, and identification was done using cellsurface antigens. Bone marrow mesenchymal stem cells of passage 3 were induced by chondrocyte induction medium to differentiate into chondrocytes (experimental group). cells cultured in normal medium were considered as control group. 3-(4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide assay was used to measure growth curve of chondrocytes. cells underwent typeⅡcol agen immunohistochemistry and toluidine blue staining. The coculture of bone marrow mesenchymal stem cells at passage 3 and Bio-gide col agen membrane were observed under an inverted phase contrast microscope and scanning electron microscope. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells with high purity and high viability were obtained by whole bone marrow blood centrifugation method and adherence screening method. cells grew wel and had strong amplified ability, and successful y differentiated into chondrocytes. Numerous bone marrow mesenchymal stem cells adhered on the Bio-gide col agen membrane, showing a tendency of multi-layer growth. cells and Bio-gide col agen membrane seem to blend into an integrant part. Cel processes appeared and connected each other and gradual y wrapped the Bio-gide col agen membrane, with the presence of obvious cel matrix secretion. These results suggested that bone marrow mesenchymal stem cells can grow and differentiate into chondrocytes on the Bio-gide col agen membrane.
4.Mechanism of metformin decreasing the cholesterol and triglyceride contents in the liver of high-fat diet fed rats
Fei WANG ; Yi LIU ; Guanqi GAO ; Hua GUO ; Bin CUI ; Ling GAO ; Jiajun ZHAO
Chinese Journal of Endocrinology and Metabolism 2008;24(2):204-207
Objective To detect the expression and activity of AMP-activated protein kinase α subunit (AMPKα) and peroxisome proliferator-activated receptor-α (PPARα) in liver of high-fat fed rats treated with metformin, and to investigate the mechanisms underlying metformin decreasing the total cholesterol (TC) and triglyceride (TG) contents of the liver. Methods Total 30 male Wistar rats were randomly divided into control group (group C), high-fat diet fed group (group HF) and high-fat diet feeding plus metformin treatment group (group Met,metformin was administered orally at the last month of high-fat diet feeding). After feeding for 5 months, TC and TG in liver and sera were determined, respectively. The mRNA and protein levels and activity of AMPKα and PPARα in the liver were determined by real-time PCR and Western blotting. The activity of PPARα transcriptor binding to DNA was detected by ELISA. Results Five months of high-fat diet feeding induced a significant decrease in AMPKα and phosphorylated-AMPKα protein expression as well as AMPKα2 and PPARα mRNA levels in the liver of rats (all P<0.05), while it did not alter PPARα protein expresssion and the PPARα activity binding to DNA as well as AMPKα1 mRNA levels. The TC and TG contents in the liver (P<0.05) and serum (P<0.05) were sharply increased in group HF than those in group C. Treatment with metformin for 1 month led to a marked increase of AMPKα2 mRNA level, AMPKα and phosphorylated-AMPKα protein expression as well as the PPARα activity in group Met compared with group HF(all P<0.05), while the PPARα protein expression and the PPARα mRNA level did not show significant change. Consistent with these findings, the TC and TG contents in rat liver as well as sera were strikingly decreased (all P<0.05). Conclusion The activations of AMPKα and PPARα induced by metformin may contribute to the decrease of TC and TG content in liver and sera of the high-fat fed rats.
5.Electrospun materials applied in drug delivery system
Zhifeng ZHAO ; Zhixun YANG ; Feng LI ; Hamushan MUSHA ; Yifan SHI ; Jian FEI ; Jun ZHANG ; Wenguo CUI
Chinese Journal of Tissue Engineering Research 2017;21(2):303-308
BACKGROUND:Recently, electrospun materials have been extensively applied in the drug delivery system. OBJECTIVE:To overview the application prospect of electrospun materials in drug delivery systems. METHODS:A computer-based search of PubMed and NCBI databases was performed for literatures about the research progress of electrospinning in tissue engineering and chemotherapy published within the past 10 years using the keywords of“electrospinning, drug delivery system, nanofibers, electrospun materials”.RESULTS AND CONCLUSION:Compared with traditional materials, electrospun stents hold good versatility and control able parameters, thus granting its unique advantage under various physiological conditions. Current drug-loaded materials composed of natural products, synthetic polymers and blended materials;as to drugs, there are antibiotics, chemotherapy medication, DNA and protein. Electrospun materials have been used in tissue engineering, cancer chemotherapy and wound healing. We focus on not only the application progress of electrospun materials in traditional treatments, but also its usage, condition-control ed drug release and living-cel carrying. Electrospun materials combined with various drug-loaded present a broad prospect.
6.Influence of intensity-modulated radiation therapy on parotid function in nasopharyngeal carcinoma
Tiantian CUI ; Shaoxiong WU ; Fei HAN ; Lixia LU ; Shaomin HUANG ; Xiaowu DENG ; Taixiang LU ; Chong ZHAO
Chinese Journal of Radiation Oncology 2009;18(3):167-169
Objective To evaluate the effect of intensity-modulated radiation therapy(IMRT) on parotid function in nasopharyngeal carcinoma(NPC). Methods Eighty-three NPC patients received prima-ry IMRT between 2001 and 2003. Xerostomia before radiotherapy, at the end of radiotherapy, at 6-month, 1-,2-,3-,4- and 5-year after radiotherapy were investigated, respectively. The relation between xerostomia and parotid dose distribution was analyzed. Results Of all the patients,4,31,31 and 17 had stage Ⅰ,Ⅱ,Ⅲ and ⅣA disease, respectively. Sixteen patients received chemo-radiotherapy. The median followed-up time was 65 months. The 5-year local control and regional control rate were 96% and 95% ,respectively. The 5-year overall survival rate was 80%. The mild xerostomia rate at the seven time points was 42%, 51%, 71%, 77%, 58%, 38% and 26%. The corresponding moderate xerostomia rate was 52%, 53%, 21%,8%, 3%, 2% and 2%, respectively. No serious xerostomia was observed. The mean dose of the bilateral parotid glands was 34.34 Gy. Xerostomia at 6-month after radiotherapy was positively correlated with the mean dose of the parotid glands, and D50 was the independent factor in predicting the xerostomia. Parotid function was well protected when the mean dose and D50 were no more than 33 Gy and 29 Gy,respectively. Conclusions IMRT can improve the local-regional control of NPC and protect the parotid glands from radiation-induced in-jury.
7. Chemical relationship study of aconitum soongaricum stapf based on HPLC fingerprint
Chinese Pharmaceutical Journal 2014;49(24):2159-2164
OBJECTIVE: To compare the HPLC fingerprints Aconitum soongaricum Stapf. with Aconitum carmichaelii Debx., Aconitum kusnezoffii Reichb., and another common radix aconite plant in Xinjiang, Aconitum leucostomum Worosch., and analyze the degree similarity between different fingerprints. METHODS: The HPLC fingerprint Aconitum soongaricum Stapf was established, and similarity analysis, cluster analysis and principal component analysis were carried out for the common patterns, Aconitum soongaricum Stapf and Aconitum carmichaelii Debx., Aconitum kusnezoffii Reichb., and Aconitum leucostomum Worosch. via the ChemPattem chemical fingerprint analysis system software solutions. RESULTS: The analysis showed that the similarities the mutual model reference fingerprints were higher between Aconitum soongaricum Stapf and Aconitum kusnezoffii Reichb. and between Aconitum leucostomum Worosch. and Aconitum carmichaelii Debx. CONCLUSION: It is preliminarily determined that Aconitum soongaricum Stapf in Xinjiang had higher similarity with the legal standard Aconitum kusnezoffii Reichb., which deserves further research and development.
8.Effect of moxibustion at Shenshu (BL 23) on the ethology, corticosterone and glucocorticoid receptor in aging rats
Fei LLEI ; Wenbin SHEN ; Yunhua CUI ; Huangan WU ; Luyi WU ; Huirong LIU ; Chunhui BAO ; Jimeng ZHAO ; Cili ZHOU ; Chen ZHAO
Journal of Acupuncture and Tuina Science 2016;14(4):250-256
Objective:To observe the effect of moxibustion on learning and memory abilities, corticosterone and glucocorticoid receptor (GR) in subacute aging rats. Methods:Twenty four Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group and a moxibustion group, 8 rats in each group. Rats in the model group and the moxibustion group were subcutaneously injected with 25% D-galactose [125 mg/(kg·bw)] for 40 d continuous; rats in the normal group were injected with saline at the same position for 40 d continuous. Rats in the moxibustion group were given mild moxibustion at bilateral Shenshu (BL 23) at the same time of modeling; rats in the normal group and the model group were only identically grabbed without moxibustion for 40 d. The learning and memory abilities of rats were observed using the Morris water maze at the end of the experiment. Abdominal aorta blood and thymus were collected after water maze experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum corticosterone level, and immunohistochemical method was used to detect the expression of thymus GR. Results:Compared with the normal group, rats in the model group showed that a significantly longer escape latency time (P<0.01) on the third and the fourth days; number of times crossing the platform in 70 s significantly reduced (P<0.01); activity times in the fourth quadrant significantly decreased (P<0.05); serum corticosterone levels increased (P<0.01); thymus GR expression decreased (P<0.05). Compared with the model group, rats in the moxibustion group showed that the escape latency times were significantly shorter on the third, the fourth and the fifth days (P<0.01,P<0.05); number of times crossing the platform in 70 s significantly increased (P<0.05); activity times in the fourth quadrant significantly increased (P<0.05); serum corticosterone levels decreased (P<0.05); thymus GR expression increased (P<0.05). Conclusion:Moxibustion could improve the learning and memory abilities of subacute aging rats, down-regulate serum corticosterone levels, and increase thymus GR content.
9.Detection and clinical significance of EGFR and KRAS mutation in peripheral blood from tumor patients by REDE-DHPLC
Zhuo YANG ; Meijuan LONG ; Fei WANG ; Qian CHEN ; Baojian ZHAO ; Ye GUO ; Yuan HUANG ; Xiulan SU ; Xu ZHANG ; Wei CUI
Chinese Journal of Laboratory Medicine 2011;34(4):327-332
Objective To establish a REDE-DHPLC method for detecting the EGFR and KRAS mutations in plasma DNA from tumor patients, and investigate its clinical significance. Methods Restriction endonucleases Mse Ⅰ , Msc Ⅰ , BstN Ⅰ and Bgl Ⅰ were used to digest the wild type fragments of exon 19,exon 21 of EGFR gene and coden 12, 13 of KRAS gene for enriching the mutation fragments, and REDE-DHPLC method was established to detect EGFR and KRAS mutations. The sensitivities of REDE-DHPLC and conventional DHPLC were analyzed by using a series of plasmids containing 50%, 10%, 5%, 1% and 0. 1% mutation genes. Then, Plasma samples and paraffin-embedded tissue samples of 120 NSCLC patients and 120 colorectal cancer patients were detected by REDE-DHPLC. Compared with conventional DHPLC and sequencing, the diagnostic efficiency of REDE-DHPLC method was evaluated by detecting the mutation status of 2 genes in plasma of NSCLC and colorectal cancer patients. Results The sensitivity values of REDE-DHPLC and conventional DHPLC for detecting mutations in 4 loci were 0. 1% and 1%respectively. Plasmid DNA containing 0.1% mutation gene was detected to be positive continually for 2 to 3 times by REDE-DHPLC. EGFR mutation rates of 120 plasma from NSCLC patients detected by REDE-DHPLC, conventional DHPLC and sequencing methods were 27. 5%, 16. 7% and 12.5% respectively, and KRAS mutation rates of 120 plasma from colorectal cancer patients were 38. 3%, 25. 8% and 16. 7%,respectively. The positive rates of EGFR and KRAS mutation detected by REDE-DHPLC were significantly higher than conventional DHPLC(x2 = 4. 092, 4. 301, all P < 0. 05 ) and sequencing method (x2= 8. 438,14. 127,all P < 0. 05 ). In comparison with conventional DHPLC, the sensitivities of REDE-DHPLC for detecting EGFR and KRAS mutation were 100% (20/20,31/31), the specificities were 87. 0% (87/100)and 83. 2% (74/89). In comparison with sequencing method, the sensitivities of REDE-DHPLC were 100%( 15/15,20/20), the specificities were 82.9% (87/105)and 74. 0% (74/100). The coincidence rate of the two methods for detecting EGFR and KRAS mutation were 89. 2% ( 107/120, Kappa = 0. 690, P < 0. 05 ) and 87.5% ( 105/120, Kappa= 0. 718, P < 0. 05 ). The Consistency of EGFR and KRAS mutation status in plasma and tissues detected by REDE-DHPLC were 91.7% (33/36, Kappa =0. 939,P <0. 05)and 90. 2 %(46/51, Kappa = 0. 914, P < 0. 05 ), respectively. Conclusions The REDE-DHPLC method is highly sensitive and specific for detecting EGFR and KRAS mutations in plasma DNA from tumor patients. The results are easy to be interpreted without missing homozygous point mutation, which indicate that the detection of EGFR and KRAS mutations in plasma DNA by REDE-DHPLC could therefore extend to be usedin clinical laboratory.
10.Magnetic resonance imaging of eyes filled with silicone oil
Qing-Hua CHEN ; Xun YANG ; Fei YAN ; Zhen-Chang WANG ; Bo ZHAO ; Ya-Jian TONG ; Ying CUI ;
Ophthalmology in China 2006;0(05):-
Objective To evaluate values of magnetic resonance imaging(MRI)in eyes filled with silicone oil.Design Prospective cases series.Participants 40 eyes of 40 patients were filled with silicone oil after ocular injury.Methods MRI was performed in the 40 patients,including axial FSE T_1WI,T_2WI,coronal T_2WI with fat saturation,oblique sagittal T_1WI and axial T_2FLAIR.MRI findings,in- cluding morpbous,signal and complications were analyzed.Oculi axes were measured.Main Outcome Measures Morphous,signal, complications and oculi axes of the eyes filled with silicone oil.Results Affected oculi axis was 2.18cm?0.21cm,normal oculi axis was 2.48cm?0.16cm.The silicone oil in eyes demonstrated isointense signal or slightly hyperintense signal on T_1WI and T_2WI,hypointense signal after fat saturation.Hydrops was found in vitreous cavity in 33 patients,including simple hydrops in 17 patients and complicated other abnormality in 16 patients.Choroidal detachment was found in 11 patients,complicating vitreous hydrops in 5 patients and lo- calized bulge of eyeball wall.Retinal detachments were found in 4 patients,of whom 3 patients complicated with vitreous hydrops.Per- fluorocarbon liquid residual in vitreous cavity,foreign body in anterior chamber,localized thickness of the wall of the globe and meagre- mean of silicone oil in vitreous cavity were found respectively in one patient complicating vitreous hydrops.Except for eye changes, fracture of orbital wall and foreign body in orbit were found in one patient.Conclusions MRI can display the changes of eyes filled with silicone oil,and measure oculi axes biologically and accurately offering important clinical application value.(Ophthalmol CHN,2007,16: 312-315)