1.Engineered Protein Nanopore for Real-time Monitoring Single-molecule Reaction between Cadmium Ion and Glutathione
Sha ZHUO ; Ying WANG ; Xiaofeng KANG
Chinese Journal of Analytical Chemistry 2017;45(8):1172-1179
The chelating reaction between glutathione peptides and divalent cadmium ion was used as a typical model for investigating the coordination chemistry of SH-containing peptides and heavy metal ions, which was essential to understand the mechanism of intracellular cadmium detoxification.Here, the mutant (CM113R)7 αHL protein nanopore equipped with a first synthesized per-6-quaternary ammonium-β-cyclodextrin (p-QABCD) was used as nanoreactor and detector to investigate the single-molecule reaction of GSH molecules and Cd2+ ions.Different reaction pathways, intermediates, and products could be recognized.Cd2+-GSH reaction was highly dependent on the solution pH.The Cd(GSH)2 was formed at pH 7.4, while the Cd(GSH)2 and Cd2(GSH)2 were formed at pH 9.0.Cd2(GSH)2 was formed by two possible pathways: (1) A Cd2+ ion primarily coordinated with the thiol group of two GSH molecules to form Cd(GSH)2, and then the second Cd2+ ion quickly incorporated with the deprotonated amino group of Cd(GSH)2 to form Cd2(GSH)2;(2) Two Cd2+ ions separately coordinated with the thiol and deprotonated amino group of one GSH molecule to form Cd2(GSH)1, and the second GSH molecule quickly bounded to Cd2+ ions to form Cd2(GSH)2.This work studied the chelating reaction between metal ions and bioactive glutathione at single-molecule level without labeling and chemical modification, and would be important to further understand intracellular mechanisms of detoxification of heavy metals and greatly expand the research field of nanopore single-molecule technique.
2.Assessment of the Function of Disabled Athletes Using Disability Assessment Schedule Ⅱ(WHO-DAS Ⅱ)
Chinese Journal of Rehabilitation Theory and Practice 2006;12(3):268-269
ObjectiveTo evaluate the reliability and validity of the of Disability Assessment Schedule Ⅱ of World Health Organization(WHO-DASⅡ) applied in assessment of the function of disabled athletes.Methods56 disabled people,26 wheelchair basketball athletes and 30 patients,were sampled and completed WHO-DASⅡ questionnaire.The test and re-test had been administrated in 10 days.Barthel Index(BI) and Sports Classification Test had been implemented for the functioning assessment.ResultsThe test-retest reliability for WHO-DASⅡ in six dimensions were: 0.978,0.807,0.938,0.877,0.998,0.935,0.986 and all were in very significant level(P<0.01).There were negative correlation between WHO-DASⅡ scores and BI in total and in every dimension,the correlation coefficients were:-0.77,-0.17,-0.71,-0.82,-0.33,-0.73,-0.61.The correlation coefficients between WHO-DASⅡ total and six dimensional scores and the grade of sports classification were:-0.741,-0.378,-0.806,(-0.541),-0.293,-0.510,-0.677.ConclusionWHO-DASⅡ can be used in assessing the function of disabled athletes.
3.Electrochemiluminescence Biosensor Based on DNAzyme and 3,4,9,10-Perylenetetracarboxylic Dianhydride Derivative Functionalized Hollow Gold Nanoparticles for Detection of Lead Ions
Xue LI ; Anyi CHEN ; Ying ZHUO ; Ruo YUAN
Chinese Journal of Analytical Chemistry 2015;(11):1701-1707
Based on target cycling amplification and 3 ,4 ,9 ,10-perylenetetracarboxylic dianhydride derivative functionalized singal probe, an ultrasensitive electrochemiluminescence ( ECL) sensor was designed for the detection of lead ions. The hairpin substrate DNA was immobilized on the electrode through molecular self-assembly. In the presence of Pb2+and DNAzyme, the substrate was cleaved with single strand DNA fragments left on the electrode surface. Meanwhile, the target and DNAzyme was released for another cleaving circularly. As a result, the single strand DNA fragments hybridized with the assist hairpin probe H1, which leaded to the fabrication of H2 labeled with the 3 , 4 , 9 , 10-perylenetetracarboxylic dianhydride derivative functionalized hollow gold nanoparticles. With the increasing concentration of Pb2+, much more signal probe was been captured and the ECL signal of the biosensor in peroxydisulfate ( S2 O2-8 ) solution would increase. An ECL assay demonstrates that the sensor has a good linear response to Pb2+ concentration in the range of 1í10-12 mol/L-1í10-6 mol/L, with a detection limit of 1í10-12 mol/L. The fabricated sensor shows good selectivity toward Pb2+against other common metal ions.
4.Comparison of bolus infusion and replenishment in contrast-enhanced ultrasound in assessing renal cortical blood perfusion
Fen HE ; Zhongxiong ZHUO ; Ying HE ; Gong WAN ; Weihua TAN
Chinese Journal of Ultrasonography 2012;21(3):249-252
Objective To compare bolus infusion and replenishment using real low mechanical index contrast enhanced ultrasound in assessing the change of renal cortical perfusion.Methods Using dopamine (i.v.) at the dose of 0.5,2.0,5.0μg · kg- 1 · min- 1 to change renal blood perfusion of 20 rabbits,then during bolus or contant injection of SonoVue,at coded pulse inversion mode,real-time contrast ultrasound was performed,the latter method needed destroying microbubble at a high MI when amplitude reach a steady state,then recording the replenishment,peak intensity(A) and time to peak (PPT) were obtained through raw time-intensity curve,and slope rate of TIC(k) was acquired by curve fitting,standard effective renal plasma flow(ERPF) was measured through 4-aminohippuric acid clearancerate method,meanwhile correlations between ERPF and parameters were analyzed,as well as the paired samples t test for each parameter before and after dopamine administration.Results The ascending branchs of raw TIC of bolus infusion increased sharply and were approximately straight,then descended gradually,while that of replenishment looked like two straightlines with different slopes,then stayed horizontal Both the value of A of two methods were positively correlated with ERPF ( r b =0.85,r re =0.66),and were different at the same ERPF,meanwhile the value of TTP were negatively correlated with ERPF( r b =-0.92,r re =- 0.76),and there were no statistically difference between the two methods.k from Gamma fitting was far from correct,while k from exponential fitting was apparently correlated with ERPF ( r re =0.77 ).Conclusions Both bolus injection and constant injection-replenishment method can assess renal cortical blood perfusion,TIC parameters A and TTP represent regional blood volume fraction and microbubble velocity respectively.Bolus-infusion with real low mechanical index is more precise and available.Comparing with k,TTP is more appropriate to reflect perfusion velocity.
5.Parameters optimization of ultrasound mediated gene transfection in HepG2
Ying HE ; Zhongxiong ZHUO ; Gong WANG ; Weihua TAN ; Fen HE
Chinese Journal of Ultrasonography 2013;(4):344-348
Objective To investigate the feasibility of transfecting gene into HepG2 by therapeutic ultrasound combined with microbubble,and to explore the optimal ultrasonic parameters for high transfection efficiency.Methods HepG2 cells were cross-interval planted in 24-well plates.EGFP plasmid DNA and microbubbles were added to the cultured HepG2 and three parameters of the therapeutic ultrasound were optimized.Firstly,set ultrasonic intensity gradient (group A,from A1 to A5),which were 0.4 W/cm,0.8 W/cm2,1.2 W/cm2,1.6 W/cm2 and 2.0 W/cm2 respectively.Secondly,set the duty cycle gradient group (group B,from B1 to B3),which were 10%,20% and 50% respectively.Lastly,set the exposure time group(group C,from C1 to C3),which were 30 s,90 s and 180 s.After 24 hours,the expression of GFP was observed by fluorescence microscopy,the transfection rates was detected by flow cytometry,the cell death were assessed by trypan blue exclusion.Results The plasmid transfection efficiency was varied under different ultrasonic parameters.Within a certain range,the increasing of parameters can improve the transfection efficiency,but when the parameters were too large (eg.the intensity>1.6 W/cm2 or 50% duty cycle),the actual transfection efficiency decreased due to the increased cell death.When the ultrasonic intensity was 1.2 W/cm2,the transfection efficiency and mortality were better than the other subgroups of group A.When the duty cycle was 20%,the transfection efficiency and mortality rate were better than the other subgroups of group B.Continue to increase the exposure time over 90 s was not statistically significant (P >0.05) for transfection efficiency and cell mortality.Conclusions The ultrasound-targeted microbubble destruction is an efficient method for gene delivery in vitro.The transfection efficiency vary greatly under different parameters,so optimization parameters is conducive to the promotion of gene transfection.
6.HPLC fingerprint of Stellera chamaejasme of Inner Mongolia Region
Zhaolian ZHUO ; Ying GAO ; Weimin LI ; Weidong GAO ; Xiaohua YAO
Chinese Traditional Patent Medicine 1992;0(06):-
AIM:To establish a HPLC fingerprint of Stellera chamaejasme L. from Inner Mongolia Region. METHODS: The RP-HPLC method was used with Akzonobel Kromasil C_ 18 (250 mm?4.6 mm, 5 ?m) the acetonitrile-0.5% phosphoric acid (gradient elution) was used as mobile phase, analytic time was 60 min, and detective wavelength was at 297 nm, the column temperature of 15℃ were adopted. RESULTS: The HPLC fingerprint of Stellera chamaejasme L. set up showed that 14 peaks were co-possessing in different sources. The results of method validation met technical standard of fingerprint, the similarities of Stellera chamaejasme L. were 0.9 to 1.0. CONCLUSION: The method is stable and reliable with a good reproducibility and provides a reference standard for the quality control of Stellera chamaejasme L. from Inner Mongolia Region.
7.Development of Medical Information Platform Based on B/W/S Structure in Clinical Department
Jun YING ; Guangfei CHEN ; Shilin HE ; Huichang ZHUO
Chinese Medical Equipment Journal 1993;0(06):-
Objective To study a feasible technology plan for information communication between doctors and patients.Methods Based on three-layer B/W/S and Windows2003 Server,MS SQL Server was used for database administration.The Apache Tomcat 5.5 was adopted as Web server and Web services were realized by using JSP(Java Server Page).Results Health care information was collected and integrated in medical information platform of clinical department which was connected into Internet directly.The information platform established communication way between doctors and patients and it had functions for chief complaint,diagnosis,reservation and revisit.Conclusion Communication between doctors and patients are realized.
8.Clinical and Pathological Analysis on 66 Cases of Neonatal Pulmonary Hemorrhage
xue, XIAO ; mei-ying, ZHUO ; you-xiang, ZHANG ; li, YU
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To explore the higher dangerous factors,the early clinical performances and its contents of neonatal pulmonary hemorrhage(NPH).Methods The clinical performances,chest radiograms and autoptical pathological materials of 66 cases of newborns who died of NPH at our neonatal department during 1993 to 2003 were reviewed and analyzed.Results The higher dangerous factors of NPH were premature delivery/low birth weight,serious diseases lead to hypoxia and severe infections.The early clinical performances of NPH were the suddenly aggravation of dyspnea and the increasing of moist sounds.The early X-ray performances were lower penetrance of lung fields extensively and well-distributly with path clouds,the intercostals space usually increased.According to the autoptical(patho)-logy,this X-ray perfomance indicated the edema of the pulmonary with small amount of hemorrhage.Conclusion The patients with the higher dangerous factors and the early clinical performances of NPH,must be diagnosed and interfered it as early as possible to reduce the mortality of NPH.
9.Advances in Application of Blue Laser Imaging in Diagnosis of Early Upper Gastrointestinal Cancer
Ying ZHUO ; Haifeng KANG ; Junbo QIAN ; Jianwei QIU ; Hongbin LIU
Chinese Journal of Gastroenterology 2017;22(6):377-380
Blue laser imaging (BLI) is a new endoscopic system equipped with the laser beam emitting two different wavelengths.It produces bright and high resolution images for observation of microvascular and microsurface patterns of esophageal and gastric mucosa, helping the diagnosis of early upper gastrointestinal cancer.Compared with the existed endoscopic techniques, BLI shows its unique advantages.In this article, advances in application of BLI in diagnosis of early upper gastrointestinal cancer were reviewed.
10.Dose-effect research using nanopatch to deliver siRNA in the inhibition of HPV gene expression.
Zhuo XIONG ; Xiaojing DONG ; Peiwen SUN ; Ying ZHANG
Journal of Biomedical Engineering 2013;30(6):1283-1289
Hela is the cell line of adenocarcinoma of the uterine cervix, and human papillomavirus (HPV) 18 shows positive. We delivered siRNA with target specifically to HPV18 E7 mRNA into nude mice Hela tumor xenografts by nanopatch to inhibit the HPV gene expression, and further to study the superiority, the best action time and concentration of siRNA of using nanopatch to transfer siRNA in vivo. We designed siRNA that target specifically to HPV18 E7 mRNA (siE7) and checked the effect of siE7 in vitro. Tumor xenografts were transfected with siE7 and GenEscort III by nanopatch. Expression of HPV18 E7 mRNA and protein were detected 0 hours, 24 hours, 48 hours, 72 hours after transfection with PT-PCR and Western blot, and the best action time was analyzed using nanopatch to thansfect siRNA in vivo. We transfected GenEscort III and siE7 of Different concentration into tumor xenografts respectively by nanopatch and intraperitoneal injection. Expression of HPV18 E7 mRNA and protein was detected 72 hours after transfection by PT-PCR and Western blot, to analyze the best action concentration of siRNA and the superiority of using nanopatch to thansfect siRNA in vivo. The results proved that SiE7was efficient to inhibit expression of HPV18 E7 mRNA and to advance Hela apoptosis in vitro. SiE7 transfected by nanopatch into xenografts could inhibit effectively expression of HPV18 E7 mRNA and protein. The best action time and concentration of siRNA of using nanopatch to thansfect siRNA in vivo are 72 hour post-transfection and 2 micromol/L siE7. To compare intraperitoneal injection in delivering siRNA in vivo, the effect of nanopatch is very predominant. It can be well concluded that Nanopatch can effectively transfer siRNA in vivo, which can effectively inhibit the HPV gene expression.
Animals
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Apoptosis
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DNA-Binding Proteins
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genetics
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Female
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Gene Expression Regulation, Viral
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HeLa Cells
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Humans
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Mice
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Mice, Nude
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Nanostructures
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administration & dosage
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Oncogene Proteins, Viral
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genetics
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Papillomaviridae
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genetics
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RNA, Small Interfering
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administration & dosage
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Transfection
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Uterine Cervical Neoplasms
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therapy
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Xenograft Model Antitumor Assays