2.The establishment of a high resolution melting analysis based rapid screening system for UGT1A1 gene mutation
Hui YANG ; Liye YANG ; Lei ZHENG ; Zhen CAI
Chinese Journal of Laboratory Medicine 2017;40(2):101-104
Objective To develop a high-resolution melting ( HRM ) assay for rapidly screening Gilbert syndrome ( GS) and Crigler-Najjar syndrome ( CNS) associated with UGT1A1 defects.Method Methodology was developed .Then, we applied the established method to analyze 61 clinical samples from neonatal patients with severe unexplained unconjugated hyperbilirubinemia .Neonates with known risk factors for developing hyperbilirubinemia , such as ABO hemolysis, G6PD deficiency, sepsis, hypoxic ischemic encephalopathy were excluded .Five pairs of PCR primers were designed to detect the five common mutations (G211A, C686A, C1091T, C1352T and T1456G) in Asia population.PCR and HRM Assay conditions were optimized.UGT1A1 genotyping in clinical samples was performed by using the established HRM analysis , and all results were subsequently confirmed by direct DNA sequencing .Results The mutants were readily differentiated by using HRM analysis .In this study, 42 neonates were identified with UGT1A1 mutation, and 4 different known variants were detected .Conclusion HRM analysis in this study was economical, convenient, rapid, effective for screening UGT1A1 gene mutations, which can serve as an reliable method for the clinical diagnosis of GS and CNS and the large-scale molecular epidemiological research of UGT1A1 gene-related diseases.
3.Analysis of Hemorheological Examination of 3483 Healthy Adults in Xi'an
Yong-ying LIU ; Ji-qing YANG ; Hui WANG ; Zhen XIANG
Chinese Journal of Rehabilitation Theory and Practice 2006;12(8):721-722
ObjectiveTo analyze the relationship between result of hemorheology and non-pathological factors such as sex, age and life habit.Methods3483 healthy adults who had health examination were divided into different groups according to sex and age, and results of hemorheological test of them were analyzed and compared with reference values.ResultsAll hemorheological indexes of men were higher than that of women. The whole blood viscosity of female had an increasing trend along with the age increasing. However, the result of hemorheology of male showed that the index of the age of 30~49 was higher than the age of more than 50, and had a decreasing trend along with the age increasing after the age of 50. The index of high shear viscosity, low shear viscosity and hematocrit of both male and female were all higher than the reference values offered by apparatus.ConclusionEffect of non-pathological factors such as age, sex and life habit on index of hemorheology should be considered.
4.Advances and Perspectives on Genetic Modification of Hevea brasiliensis
Zhi ZOU ; Li-Fu YANG ; Zhen-Hui WANG ; Kun YUAN ;
China Biotechnology 2006;0(01):-
As the major commercial source of natural rubber,Hevea brasiliensis attracts much attention.However,the heterozygous nature,long breeding cycle are strong limitations for conventional breeding.While genetic engineering,which can be used to widen the germplasm base and produce desirable agronomic traits quickly and efficiently,offers a viable alternative approach to complement traditional breeding.Comprehensive analysis indicates that in the past two decades,with calli derived from immature anther or integumental tissues of immature fruit as receptors,both biolistic and Agrobacterium-mediated transformation methods were employed for developping rubber genotypes with improved latex yield,tolerance to tapping panel dryness syndrome,producing high-value recombinant proteins,etc.Being recalcitrant to tissue culture,the transformation efficiency of Hevea is comparatively low,and the procedures are still needed to optimize.Finally,breeding objectives and strategies to improve transformation efficiency were also proposed in the review.
5.UPLC-MS/MS determination of content of three iridoids of xingnaojing oral preparation in rat brains and study on their brain pharmacokinetics.
Pan XU ; Shou-Ying DU ; Yang LU ; Jie BAI ; Hui-Min LIU ; Qiu DU ; Zhen-Zhen CHEN ; Zhen WANG
China Journal of Chinese Materia Medica 2014;39(12):2351-2355
To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.
Animals
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Brain
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metabolism
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Brain Chemistry
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Chromatography, High Pressure Liquid
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methods
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Drug Stability
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Drugs, Chinese Herbal
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chemistry
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pharmacokinetics
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Iridoids
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chemistry
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pharmacokinetics
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Male
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Rats
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Rats, Sprague-Dawley
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Tandem Mass Spectrometry
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methods
6.Predictive value of (99)Tc(m)-MIBI scintimammography in evaluation of the efficacy of neoadjuvant chemotherapy in patients with operable breast cancer.
Zhen-zhen LIU ; Zhen-duo LU ; Heng-wei ZHANG ; Hui YANG ; Hui LIU ; Lian-fang LI ; Wen-Liang LI ; Shu-de CUI
Chinese Journal of Oncology 2011;33(7):544-546
OBJECTIVETo investigate the value of technetium-99m methoxyisobutylisonitrile ((99)Tc(m)-MIBI) imaging in predicting the efficacy of neoadjuvant chemotherapy (NCT) and prognosis in patients with operable breast cancer.
METHODSSixty five patients with breast cancer underwent (99)Tc(m)-MIBI scintimammography before NCT, and static planar images were taken at 10 min and 180 min after scintimammography. The clearance rate was calculated in each patient, correlation between the clearance rate and efficacy of NCT, and the disease free survival rate were analyzed.
RESULTSThe mean clearance rate of 65 patients was (17.4 ± 6.8)%. The efficacy of NCT was 86.2% (CR 4 cases, PR 52 cases, SD 8 cases, and PD 1 case), and the mean clearance rate of patients with good response or poor response of chemotherapy were (15.5 ± 5.0)% and (29.2 ± 3.2)%, respectively. There was a significant difference between the two groups. The average disease free survival rate in the group with low clearance rate was (75.8%, P = 0.046), significantly higher than that in the group with high clearance rate (53.1%).
CONCLUSIONScintimammography of (99)Tc(m)-MIBI may be used to evaluate the efficacy and prognosis of NCT for patients with operable breast cancer.
Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; diagnostic imaging ; drug therapy ; Carcinoma, Ductal, Breast ; diagnostic imaging ; drug therapy ; Carcinoma, Lobular ; diagnostic imaging ; drug therapy ; Chemotherapy, Adjuvant ; Cyclophosphamide ; therapeutic use ; Disease-Free Survival ; Epirubicin ; therapeutic use ; Etoposide ; therapeutic use ; Female ; Fluorouracil ; therapeutic use ; Follow-Up Studies ; Humans ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Predictive Value of Tests ; Radionuclide Imaging ; Radiopharmaceuticals ; Remission Induction ; Taxoids ; therapeutic use ; Technetium Tc 99m Sestamibi
7.Expression of stanniocalcin 1 in papillary thyroid carcinoma and its clinical significances in gerontal patients
Hui FANG ; Ying YANG ; Gang XU ; Ruhua GUAN ; Meiliu YANG ; Yanfeng ZHEN
Chinese Journal of Geriatrics 2016;35(3):301-304
Objective To investigate the protein expression of stanniocalcin 1 (STC1) in papillary thyroid carcinoma (PTC) in gerontal patients versus non-gerontal subjects,and its relationship with clinicopathological features.Methods The protein expression levels of STC1 in PTC,nodular goiter and normal thyroid tissues in the gerontal patients versus non-gerontal subjects were detected by immunohistochemistry.The protein expressions of STC1 were detected by Western blotting.Results The protein expression of STC1 in gerontal patients was higher in PTC than in nodular goiter and normal thyroid tissues (60.9% vs.30.0%,15.0%,P<0.05 or 0.01).The relative expression of STC1 protein in gerontal patients was higher in PTC than in nodular goiter and normal thyroid tissues [(0.647 ± 0.076) vs.(0.280 ± 0.039),(0.248 ± 0.065),F =9.965 and 1.143,both P<0.01].STC1 protein expression had no correlations with age,gender,tumor diameter and tumor position in patients (P<0.05),while it was associated with tumor lymph node metastasis and clinical stage (P<0.05).There was no significant difference in above indexes between the elderly and non-elderly patients with papillary thyroid carcinoma (F=0.007,P=0.934).Conclusions STC1 protein may be associated with the development of papillary thyroid carcinoma,and it may has a some reference value in differentiating benign from malignant thyroid neoplasms and in predicting the prognosis of thyroid carcinomas.
8.Rapid detection of extrinsic harmful contaminants using quantum dots-coated probes and their application prospects in traditional Chinese medicine.
Xi-Hui YANG ; Wei-Jun KONG ; Mei-Hua YANG ; Shi-Lin CHEN ; Ming ZHAO ; Zhen OUYANG
China Journal of Chinese Materia Medica 2013;38(23):3997-4005
The contamination of extrinsic harmful contaminants including mycotoxins, heavy metals and pesticides, etc, brings serious risks to traditional Chinese medicines (TCMs), further to human health. Due to their unique photoluminescence, chemiluminescence, electrochemical and electrochemiluminescence properties, semiconductor quantum dots (QDs) nanoparticles are widely used to immobilize bioprobes and biosensors, etc. In this review, the luminescence characteristics and specific ligands of QDs probles which are used to determine contaminants were summed up. Then, the applications of QDs-coated novel probes in the determination of mycotoxins, heavy metals and pesticides were discussed in detail. In addition, the contamination levels and characteristics of extrinsic harmful residues in TCMs were investigated. Further, the maximum levels of those contaminants in TCMs were compared with those set by various countries. Finally, the future development trends and problems of QDs-coated probes in the determination of those extrinsic residues in TCMs were prospected.
Drug Contamination
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prevention & control
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Humans
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Medicine, Chinese Traditional
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methods
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Nanotechnology
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instrumentation
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methods
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Quantum Dots
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Safety
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Time Factors
9.Staurospolin promotes the apoptosis of multidrug resistance tumor cell lines induced by adriamycin
Shi-Yong WANG ; Wei-Li DU ; Zhen-Jun YANG ; Huan YU ; Yun-Feng YANG ; Hui ZHANG ;
Cancer Research and Clinic 2006;0(12):-
Objective To investigate whether the protein kinase C inhibitor can promote the apopto- sis of multidrug resistance tumor cell lines which are induced by chemotherapy drugs.Methods Choose the KB/S(oral squamous cancer cell line)and KB/VCR(its multidrug resistant cell line)to compare the Adri- amycin-induced apoptosis with or without staurospolin(protein kinase C inhibitor).The apoptosis is stained with acridine orange,tested by flow cytometry,and approved by electron microscope.Results 36 hours after the treatment with 0.04 ?g/ml adriamycin,apoptotic cells of KB/S are 96.68%,and after 48 hours,the apop- totic cells of KB/VCR are 64.99%.When the concentration of adriamycin are augmented to 0.4?g/ml and 2.0?g/ml,the apoptotic cells of KB/VCR are 69.74% and 37.18% respectively.When treated with stau- rospolin together,the apoptotic cells of KB/VCR increased to 72.58%(?~2=4.5,P0.05)respectively.These results were testified by electron microscope and acridine orange-stain.Conclu- sion The resistance to apoptosis may be one of the mechanisms of multidrug resistance and the protein ki- nase C inhibitor may reverse this resistance by promoting the apoptosis of multidrug resistance tumor cells.
10.The relationship between 99mTc-MIBI scintimammography of breast cancer and multidrug-resistant proteins.
Shu-de CUI ; Zhen-zhen LIU ; Hui LIU ; Lian-fang LI ; Hui YANG ; Wen-liang LI
Chinese Journal of Oncology 2005;27(10):606-608
OBJECTIVETo evaluate the relationship between the clearance rate of technetium-99m methoxyisobutylisonitrile ((99m)Tc-MIBI) in scintimammography and multidrug-resistant proteins expression in breast cancer tissues.
METHODSSeventy-six patients with breast cancer underwent (99m)Tc-MIBI scintimammography before treatment, and static planar images were taken at 10 min and 180 min after scintimammography. The clearance rate of (99m)Tc-MIBI was calculated in each patient. Immunohistochemical analysis was performed on pathological specimens of the 76 breast tumors to determine the expression of P-glycoprotein (P-gp), glutathione S-transferase Pi (GST-pi) and topoisomerase II (Topo II).
RESULTSThe clearance rate was significantly higher in 36 patients with positive P-gp expression when compared with that in 40 patients with negative P-gp expression. There was no significant relationship between GST-pi, Topo II and the clearance rate of (99m)Tc-MIBI.
CONCLUSIONThe clearance rate of (99m)Tc-MIBI in breast imaging may be used to evaluate the P-gp level in breast cancers.
ATP Binding Cassette Transporter, Sub-Family B ; analysis ; Adult ; Aged ; Breast Neoplasms ; chemistry ; diagnostic imaging ; Carcinoma, Ductal, Breast ; chemistry ; diagnostic imaging ; DNA Topoisomerases, Type II ; analysis ; Drug Resistance, Multiple ; Female ; Glutathione S-Transferase pi ; analysis ; Humans ; Immunohistochemistry ; Middle Aged ; Multidrug Resistance-Associated Proteins ; analysis ; Radionuclide Imaging ; Radiopharmaceuticals ; Technetium Tc 99m Sestamibi