1.Quantum Dots Labeled Lateral Flow Strip for Determination of Procalcitonin in Blood
Hao YU ; Liang XU ; Xiaoping QI
Chinese Journal of Analytical Chemistry 2014;(11):1592-1597
CdSe/ZnSquantumdots(QDs)werepreparedandcovalentlylinkedtoanti-katacalcinmonoclonal antibodies. After modification, the QDs' maximum emission wavelength was shifted to 625 nm from 620 nm while maintaining the spectral properties. Then the QDs labeled lateral flow strip and corresponding fluorescence measuring instrument were designed and fabricated. To reduce the cost of strip by reducing the amounts of monoclonal antibodies, appropriate amounts of QDs labeled monoclonal antibodies were sprayed on the conjugation pad, with just one test line on the strip but without the control line. Parameters of the strip were optimized by measuring the signal to noise ratio. By using the strip and fluorescence measuring instrument, procalcitonin (PCT) could be detected in 20 min, and the quantitative detection range was 0. 2-100 μg/L with sensitivity of 0. 1 μg/L. A total of 22 blood samples were measured by both our method and the commercial instrument used in the hoptital. The results were consistent for their Pearson correlation coefficient (0. 9995) and Kolmogorov-Smirnov test (Sig=1. 0). The rapid quantitative detection method for PCT is of great importance to quantitative detection of bacterial infection and rational usage of antibiotics clinically.
2.The expression and meaning of liver X receptor in obstructive sleep apnea-hypopnea syndrome patients
Yongjie QI ; Meijuan XU ; Qin YU
Chinese Journal of Practical Internal Medicine 2001;0(07):-
Objective To study the liver X receptor(LXR)and the gene expression of cholesterol ester transfer protein(CETP),cyclooxygenase-2(COX2)in obstructive sleep apnea hypopnea syndrome(OSAHS)patients.Methods Macrophages were separated from the blood drawn in 30 cases of OSAHS patients and 30 healthy control subjects in the morning,RT-PCR gel eletrophoresis was used to determine the patterns of gene expression.Results OSAHS patients had higher LXR,COX2,CETP expression(P
3.Application of spinal navigation with the intra-operative 3D-imaging modality in vertebroplasty to treat osteoporotic vertebral compression fracture
Orthopedic Journal of China 2006;0(12):-
[Objective]To study the effect of intra-operative three-dimensional fluoroscopy-based spinal navigation guidance for vertebroplasty to treat osteoporotic vertebral compression fracture(OVCF).[Method]A clinical study was performed in which 41 patients suffering painful osteoporotic vertebral compression fractures underwent vertebroplasty procedure.During this procedure,cannulation of the pedicle and vertebral body was performed with the aid of isocentric 3D fluoroscopy-based spinal navigation(group A) and biplanar fluoroscopy(group B).RDQ and VAS of each group was compared between preoperation and postoperation.Total operating time,intra-operative fluoroscopy time and pedicle puncturation accuracy were compared between group A and group B.Possible complications such as cement extravasations were evaluated with X-ray fluoroscopy.[Result]Forty-six vertebrae were successfully injected with polymethyl methacrylate.The two groups had statistically significant difference in RDQ and VAS between preoperation and postoperation,but no statistically significant difference between the two groups.Mean operating time and intra-operative fluoroscopy times of group A were shorter than those in group B.There was no case of broken pedicle in group A.One vertebra was found to have bone cement leakage to soft tissue in group B,but the vertebral canal and pedicle intact.The author followed up for an average of 21 months(range 8~38 months) and found no severe complication or collapse of vertebra.[Conclusion]With the aid of intra-operative three-dimensional fluoroscopy-based spinal navigation,vertebroplasty can be performed more accuractly and securely.The shorter total operating time,intra-operative fluoroscopy times are convincing and less possible complication can be expected.
4.Different active ingredients of medicinal plant based on function differentiation of homologous gene.
Yuan YUAN ; Jun YU ; Lu-qi HUANG ; Xu-min WANG
China Journal of Chinese Materia Medica 2015;40(6):1023-1026
In the research field of quality control in Chinese medicinal materials, variation in active ingredients of medicinal plant is always the key and hot issues. With the development of high-throughput sequencing technologies and reducing cost, a large numbers of genes from medicinal plant were cloning and provide a solid foundation for further research of gene structure and its biological function, and also provides conditions for explore active ingredient variation and its quality control from the perspective of molecular pharmacognosy. This paper introduces the concept of homologous gene, gene duplication and classification. We prospect the function of duplicated genes in the role of molecular mechanism research about variation in active ingredients, aiming at providing a new way for medicinal materials quality control.
Drugs, Chinese Herbal
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analysis
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Gene Duplication
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Plant Proteins
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genetics
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Plants, Medicinal
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chemistry
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genetics
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Quality Control
5.Effects of transforming growth factor β1 and β3 gene transfer on MMP-2,MMP-9 and TIMP-1 expression in hepatic stellate cells in rats
Jiao YU ; Xia ZHOU ; Qi LI ; Wei QIAN ; Keshu XU
Chinese Journal of Clinical Infectious Diseases 2008;1(3):159-162
Objective To investigate the effects of transforming growth factorβ1(TGFβ1)and β3 (TGFβ3)gene transfer on MMP-2,MMP-9 and TIMP-1 expression in hepatic stellate cells(HSC-T6).Methods TGFβ1 and TGFβ3 expression plagmids were constructed.The recombinant expression plasmid pcDNA3.1(+)-=TGFβ1 and pcDNA3.1(+).TGFβ3 were transfected or cotransfected into HSC-T6.At 24,48 and 72 h after transfection,the expression of MMP-2,MMP-9 and TIMP-1 mRNA were detected by real-time quantitative PCR,and the expression of MMP-2,MMP-9 and TIMP-1 protein were detected by Western blot.The recombinant expression plasmid pcDNA3.1(+).TGFβ1 was transfected into HSC-T6,and positive clones were selected by G418.The positive clones were transfected by the recombinant expression plasmid pcDNA3.1(+).TGFβ1,and the expression of MMP-2,MMP-9 and TIMP-1 were detected at 48 h after transfection.Results After transfection with peDNA3.1-TGFβ1,MMP-2 and TIMP-1 increaged remarkably in HSC-T6 cells(P<0.05),but MMP-9 remained at the sanle level;After transfection with pcDNA3.1-TGFβ3,expression levels of MMP-2,MMP-9 and TIMP-1 mRNA were not changed,but TIMP-1 protein increased remarkably(P<0.05);in cotransfection group,the expression of MMP-2 was higher than that in the blank and the control groups(P<0.05),but MMP-9 level was not changed and TIMP-1was decreased compared with that in the TGF-β1 transfection group(P<0.05).After TGFβ3was transfected into positive clones,the change of MMP-2 wag not significant(P>0.05).but MMP-9 increaged and TIMP-1 decreased significantly at 48 h after transfection(P<0.05).Conclusions TGFB3 may inhibit liver fibrosis by increase the activity of MMP-2 and MMP-9,and decrease the activity of TIMP-1.
6.Dynamic contrast-enhanced MRI in serological negative early stage rheumatoid arthritis compared with osteoarthritis in hand
Yu WANG ; Ji QI ; Xinwei LEI ; Huiping XU ; Peng LI
Chinese Journal of Radiology 2010;44(7):721-725
Objective To investigate the value of dynamic contrast-enhanced magnetic resonance imaging ( DCE-MRI) in differentiating serological negative early stage rheumatoid arthritis (RA) from osteoarthritis (OA)in hand. Methods Plain MRI, enhanced MRI of whole hand and DCE-MRI of single slice were performed in 53 patients suspected of RA or OA and 18 healthy volunteers. Twenty-three of them were diagnosed as RA,including 18 cases of serological negative RA and 18 of them were diagnosed as OA after 3 to 6 months follow-up. The contrast rate,slope, thickening of synovial membrane were measured on DCE-MRI in three groups and the MRI findings were also were detected in both RA and OA groups. The contrast rate and slope of synovial membrane were compared among three groups using rank sum test. The thickening of synovial membrane were compared among three groups using variance analysis. MRI signs of RA and OA group were evaluated with rank sum test Results The dynamic contrast rate of synovial membrane in RA group,OA group and control group was(100.78±61.96)%,(40.44±15.43)% and (23.56±9.14)%,respectively.Individually,RA group to OA group,u=3.101,P=0.002;RA group to control group,u=4.669,P=0.000;OA group to control group,u=3.482,P=0.000.The slope of contrast curve of synodal membrane of RA group,OA group and control group was 72.50°±13.34°,45.39°±9.94°,14.56°±5.75°,respectively.Individually,RA group to OA group,u=8.002,P=0.000;RA group to control group,u=17.102,P=0.000;OA group to control group,u=9.100,P=0.000.The synovial membrane thickening of RA group,OA group and control group was(3.3±0.5),(2.8±0.7)and (1.4±0.6)mm,respectively.Individually,RA group to OA group,q=2.622,P=0.011;RA group to control group,q=9.583,P=0.000;OA group to control group,q=6.961,P=0.000.Conclusion The quantitative index of DCE-MRI,including contrast rate and synovial membrane thickening may provide useful information for differentiating OA from semlogical negative early stage RA.The contrast rate and thickening of synovial membrane in RA group are higher than those in OA group.Many signs of MRI Can help differentiate OA from RA.
7.Synthesis of Novel Hapten and Development of Monoclonal Antibody-based Enzyme-Linked Immunosorbent Assay for Malachite Green in Fish Samples
Yu WANG ; Jinyi YANG ; Zhenlin XU ; Ping QI ; Yudong SHEN
Chinese Journal of Analytical Chemistry 2016;44(9):1385-1393
To produce specific antibodies against malachite green ( MG) , one special hapten was synthesized and characterized, and conjugated to carrier protein as immunogen. The immunogen showed excellent reactogenicity and immunogenicity. One specific monoclonal antibody (mAb, named MG-DA4-C7) with high sensitivity and specificity for MG in indirect competitive enzyme-linked immunoassay ( icELISA ) was screened. The isotype was IgG1 and the light chain was κ type. After optimization of ELISA conditions, the proposed icELISA showed a 50% inhibition value ( IC50 ) of 0. 96 μg/L, a linear range ( IC20-IC80 ) of 0. 1-8. 1 μg/L and a limit of detection ( LOD, IC10 ) of 0. 05 μg/L for determination of MG. The assay showed cross-reactivity of 18. 1%, 26. 5% with crystal violet and brilliant green, respectively, and negligible cross-reactivity with other metabolites of MG (<0 . 1%) . The average recoveries of MG from spiked fish samples were from 87. 3% to 107. 3%. Good correlation (R2=0. 999) was obtained between the results of icELISA and those of liquid chromatography-tandem mass spectrometry analysis. The proposed icELISA is suitable for the determination of MG in fish samples in a simple and sensitive manner.
8.Effects of epigallocatechin gallate on reactive oxygen species in mouse podocytes exposed to high glucose
Yan DAI ; Qing YU ; Qi XU ; Jian YAO ; Weijie YUAN
Chinese Journal of Nephrology 2009;25(1):31-35
Objective To investigate the influence of epigallocatechin gallate ( EGCG)on reactive oxygen species (ROS) in mouse podocytes exposed to high glucose. Methods Mouse podocytes cultured in high glucose were exposed to different concentrations of EGCG (0.2, 10, 100 μmol/L) or α-tocopherol (0.2 μmol/L) for 6, 12, 24 hours. The viability of podocytes was detected by MTT. The intracellular formation of ROS was detected by confocal microscopy with fluorescent probe CM-H2DCFDA and was measured by fluorescence microscopy. RT-PCR was used to examine the expression of p22phox, p47phox and p67phox mRNA in cultured podocytes exposed to different concentrations of EGCG. Results Intracellular ROS generation was significantly higher in high glucose than that in control conditions (P<0.01). EGCG could significantly inhibit ROS induced by high glucose significantly (P<0.01). EGCG (100 μmol/L) led to an inhibition of the increased production of NADPH oxidase components of p22phox and p67phox mRNA in high glucose (P<0.05). The expression of p47phox mRNA in high glucose was inhibited by EGCG(0.2 μmol/L) and ot-tocopherol(0.2 μmol/L) (P<0.05). Conclusion EGCG can protect cultured mouse podocytes from injury of high glucose by inhibiting ROS formation.
9.Comparative study of 18F-FDG coincidence SPECT and 9gTcm-MDP bone scan for detecting malignancy
Yu-bo, MA ; Qi, YUAV ; Feng, XU ; Ai-chun, GU
Chinese Journal of Nuclear Medicine 2011;31(6):390-393
ObjectiveTo compare the diagnostic efficiency of 18F- FDG coincidence SPECT/CT and 99Tcm- MDP whole body bone scan (WBBS) in detecting malignancy.MethodsA total of 71 cases (male 45,female 26,mean age 59.2 ± 15.4 years) with clinically confirmed malignancy underwent both 99TcmMDP WBBS and 18F-FDG coincidence imaging within three weeks.The sensitivity,specificity,accuracy,positive and negative prediction value of these two imaging methods in detecting bone metastases were compared based on the results from pathology or clinical follow-up.x2test was used for data analysis.ResultsA total of 350 lesions (including primary,second malignancy and benign disease) in 71 patients were eval-uated.81.7% (286/350) malignant lesions were identified by either 99Tcm-MDP WBBS (209/350,59.7% ) or 18F-FDG coincidence imaging ( 141/350,40.3% ) (x2 =25.65,P < 0.01 ).The imaging findings of osteoblastic,osteolytic,mixed types of bone metastases by99Tcm-MDP WBBS and 18F-FDG coincidence imaging were significantly different (x2 =20.78,2.89 and 9.94,all P < 0.05 ).The sensitivity,specificity,accuracy,false-positive,false-negative,positive and negative predictive values for detecting bone metastases by 18 F-FDG coincidence study and 99Tcm-MDP WBBS were as follows:11.72% ( 15/128),91.67%(22/24),24.34% (37/152),8.33% (2/24),88.28% (113/128),88.24% (15/17),16.30% (22/135) ; and 53.91% (69/128),75.00% ( 18/24),57.24% (87/152),25.00% (6/24),46.09% (59/128),92.00% (69/75),23.38% ( 18/77 ).The sensitivity,accuracy,false-negative,positive-predicting value of the two methods had been significant different (x2 =32.70- 46.21,all P < 0.01 ).When two methods were combined,the diagnostic efficiency could been improved.ConclusionThe 99Tcm-MDP WBBS and 18F-FDG coincidence imaging has a complementary role in detecting bone metastases.
10.The accuracy of intravesical prostatic protrusion for diagnosing bladder outlet obstruction: A meta-analysis
Ding XU ; Tao HUANG ; Yongjiang YU ; Yang JIAO ; Jun QI
Chinese Journal of Geriatrics 2013;(1):99-102
protrusion measured by ultrasound for diagnosing bladder outlet obstruction.Methods A literature search of medline (1966.1-2011.6),embase(1984.1-2011.6),CNKI (1994.1-2011.6) and WEIPU Data (1989.1-2011.6) from 1999 to 2009 was performed by two reviewers independently.QUADAS items was applicated to assess trial quality.Golden standard was BOOI measured by urodynamics (BOOI more than 40 indicates bladder outlet obstruction).Heterogenous studies and meta-analysis were conducted by Meta-Disc 1.4 software.Results Totally 6 studies was included at last,involving 682 subjects.No threshold effect was found,but there was heterogeneity due to other factors.The meta-analysis showed that the sensitivity was 70.8 %,specificity was 87.6 %,positive LR was 5.132,negative LR was 0.303,the diagnostic OR was 22.18,the area under SROC curve was 0.8723 and Q index was 0.8028.Conclusions Intravesical prostatic protrusion measured by ultrasound is a good index for diagnosing bladder outlet obstruction in patients with benign prostate hyperplasia when intravesical prostatic protrusion is equal or more than 10mm.