1.Rapid simulation of electrode surface treatment based on Monte-Carlo model.
Zhengtian HU ; Ying XU ; Miao GUO ; Zhitong SUN ; Yan LI
Journal of Biomedical Engineering 2014;31(6):1361-1367
Micro- and integrated biosensor provides a powerful means for cell electrophysiology research. The technology of electroplating platinum black on the electrode can uprate signal-to-noise ratio and sensitivity of the sensor. For quantifying analysis of the processing method of electroplating process, this paper proposes a grid search algorithm based on the Monte-Carlo model. The paper also puts forward the operational optimization strategy, which can rapidly implement the process of large-scale nanoparticles with different particle size of dispersion (20-200 nm) attac- hing to the electrode and shortening a simulation time from average 20 hours to 0.5 hour when the test number is 10 and electrode radius is 100 microm. When the nanoparticle was in a single layer or multiple layers, the treatment uniformity and attachment rate was analyzed by using the grid search algorithm with different sizes and shapes of electrode. Simulation results showed that under ideal conditions, when the electrode radius is less than 100 /m, with the electrode size increasing, it has an obvious effect for the effective attachment and the homogeneity of nanoparticle, which is advantageous to the quantitative evaluation of electrode array's repeatability. Under the condition of the same electrode area, the best attachment is on the circular electrode compared to the attachments on the square and rectangular ones.
Algorithms
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Biosensing Techniques
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Electrodes
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Models, Theoretical
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Monte Carlo Method
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Nanoparticles
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Particle Size
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Platinum
2.Monitoring the migration of bone marrow derived mesenchymal stem cells to intracranial glioma by sodium iodide sympoter
Shuo SHI ; Min ZHANG ; Rui GUO ; Ying MIAO ; Biao LI
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;35(5):346-350
Objective To construct a recombinant lentiviral expression vector containing NIS and EGFP gene,and to explore the feasibility of NIS gene for monitoring the bone marrow derived mesenchymal stem cells (BMSCs) migration to the intracranial glioma.Methods The NIS and EGFP gene fragments were subcloned into lentiviral vector pLVX-puro,then packaged and amplified in HEK293T cells to obtain recombinant lentivirus pLVX-CMV-NIS-EGFP.pLVX-CMV-0-EGFP was constructed as control.BMSCs were isolated,cultured,and transfected by lentivirus.The antibiotic-resistant transfected BMSCs (BMSCs-NIS-EGFP and BMSCs-EGFP) were selected.The expression of NIS gene was examined by Western blot.Functional NIS activity was confirmed by the uptake of 125I and the inhibition effect of NaClO4.The nude mice intracranial glioma models were established.MicroSPECT was performed at 24 h post BMSCs-NIS-EGFP injection via the tail vein.Results pLVX-CMV-NIS-EGFP and pLVX-CMV-0-EGFP were successfully constructed and packaged.BMSCs were successfully isolated and cultured.Stable cell lines BMSCs-NIS-EGFP and BMSCs-EGFP were constructed after lentivirus transfection and puromycin selection.The expression of NIS gene was detected by Western blot in BMSCs-NIS-EGFP,but not in BMSCs-EGFP.BMSCs-NIS-EGFP showed significantly more uptake of 125I (nearly 10 times than the uptake in BMSCs-EGFP) and the uptake could be significantly inhibited by NaClO4.The nude mice intracranial glioma models were successfully established and the BMSCs-NIS-EGFP in glioma foci could be visualized by microSPECT imaging at 24 h post injection.Conclusions A recombinant lentivirus containing NIS gene could be successfully constructed for monitoring BMSCs migration towards intracranial glioma.It might provide evidence on the research of BMSCs and NIS gene mediated therapy for glioma.
3.Effect of Acupuncture at Point Neiguan on Myocardial Ultramicrostructure in Myocardial Hypertrophy Mice
Miao ZHANG ; Wenjun WANG ; Pengyu ZHU ; Ying GUO ; Shulin LI
Shanghai Journal of Acupuncture and Moxibustion 2016;35(5):588-591
Objective To investigate the effect of acupuncture at point Neiguan on mouse myocardial hypertrophy.Methods Thirty-sixty healthy C57BL6 mice were randomized into blank, model and treatment groups, 12 rats each. In the treatment group, an acupuncture needle was inserted into point Neiguan and retained for 15 min, once daily. After nine days of treatment, the ratios of cardiac weight/tibial length were compared between the groups. Left ventricular myocardial tissues were processed by Masson trichrome staining in every group of mice. The differences in myocardial fiber arrangement and collagen fiber distribution between the groups were observed under the ordinary optical microscope in 100 times of field of vision. Sarcomere structure, myofibril arrangement, collagen fibers and mitochondria in injured myocardial tissues were observed by transmission electron microscopy. Results Heart rate increased significantly in the model group and decreased significantly in the acupuncture group compared with the blank group. Cardiac weight and the ratios of cardiac weight/tibial length increased significantly in the model and acupuncture groups compared with the blank group (P<0.01) and decreased significantly in the acupuncture group compared with the model group (P<0.01). In the model group, myocardial tissues had compensatory hypertrophy with incontinuous status, and myocardial transverse striations had severe disorder/disruption, karyopyknosis appeared and cytoplasmic eosinophilic change became obvious in few of the rats. The results were better in the acupuncture group than in the model group. In the model group, part of myofibrils had focal fusion, interstitial collagen fibers proliferated and mitochondria had swelling and compensatory increase. The results were also better in the acupuncture group than in the model group.Conclusions Acupuncture at point Neiguan can reduce the production of collagen fibers and mitochondria, improve the arrangement of sarcomere structure and delay the progression of myocardial hypertrophy.
4.Construction of recombinant HIF-1α and NIS lentiviral expression plasmid and its functional identification
Shuo SHI ; Rui GUO ; Lihua WANG ; Min ZHANG ; Miao ZHANG ; Ying MIAO ; Biao LI
Chinese Journal of Nuclear Medicine and Molecular Imaging 2014;34(2):130-135
Objective To construct a recombinant lentivirus vector containing the human NIS gene and HIF-1α with the myosin light chain-2v(MLC-2v) as a promoter and to investigate the specific expression and feasibility of NIS as a reporter gene in cardiomyocytes.Methods The target gene HIF-1α and NIS were subcloned into the lentivirus (Lv)-elongation factor (EF)1-HIF-1α-internal ribosome entry site (IRES)-NIS and Lv-MLC-HIF-1α-IRES-NIS lentivirus vectors.The recombinated vectors were transfected into Hela cells by lipofectamine 2000.The expression of HIF-1α and NIS in the transfected Hela cells was detected by indirect immunofluorescence and Western blot.The H9C2 cells were exposed to different multiplicities of infection (MOI; 5,10,20,40) with packaged virus particles.The infection efficiency was detected by Western blot.MOI 20 was used for H9C2,NIH-3T3 and L6 cell lines and the specificity of the MLC-2v promoter was detected by the count of NIS protein in the 3 different cell lines with Western blot.The function and features of NIS protein were evaluated by dynamic iodine uptake and NaClO4 iodine uptake inhibition tests in vitro.Two-sample t test was used to analyze the data.Results The two recombinant lentivirus vectors were constructed successfully.The HIF-1α protein was expressed in the cytoplasm and the NIS protein was expressed on the cell membrane in Hela cells.The grey levels of NIS and HIF-1α proteins in the positive control were 69.8 and 71.9,respectively,which were 109.4 and 92.7 after being prompted by EF1,and 141.9 and 132.4 by MLC-2v.The expression of these proteins was much higher by EF1 promoter than that by MLC-2v promoter.The optimal MOI for the Lv-MLC-HIF-1α-IRES-NIS virus to infect H9C2 cells was 20.With the MOI of 20,the grey levels of NIS protein promoted by EF1 were 23.4,29.8 and 28.6 for H9C2,NIH-3T3 and L6 cells infected with Lv-EF1-HIF-1α-IRES-NIS virus,respectively.The expression of NIS protein promoted by MLC-2v was much higher in H9C2 cells than the other two cell lines.The grey level of NIS protein was 157.9 in H9C2 cells,178.8 in L6 cells and 217.3 in NIH-3T3 cells.The NIS protein expressed in infected H9C2 cells showed high radioiodine uptake.The peak of iodine uptake was 4 287.2 counts · min-1 at 40 min which was 16.85 times of the control group (254.4 counts · min-1) (t=5.34,P< 0.01).The inhibition rate of iodine uptake was up to 85.5% (3 666.4/4 287.2,t=21.3,P<0.01) by NaClO4.Conclusions MLC-2v promoter allows specific expression of the external gene HIF-1α and NIS in myocardium.The cardiomyocytes transfected with NIS gene acquires the function of iodine uptake.Therefore,NIS may have a potential to be the reporter gene to monitor the external gene therapy in ischemic cardiomyopathy.
5.Health Conditions and Health Service for People with Intellectual Disability
Miao-Miao WANG ; Guo-Hua LU ; An-Qiao LI ; Zhuo-Ying QIU
Chinese Journal of Rehabilitation Theory and Practice 2019;25(1):15-21
Objective To analyze the health conditions, related factors and development of health services for people with intellectual disability, to provide evidence-based support for the development of health services.Methods The functioning of people with intellectual disability had been analyzed using ICF. Health conditions and related factors for people with intellectual disability had been analyzed using epidemiologic approach.Results The prevalence of intellectual disability is about 1.1% reported by US and 0.75% by China. The main health conditions for people with intellectual disability were overweight, obesity, malnutrition, oral problems and digestive diseases. The main functioning included cognitive impairment, autism spectrum disorder, visual or auditory impairment, learning disability, movement limitation, generalized developmental disorder, etc. Secondary disorders caused by intellectual disability worsened the functioning of conceptual skills, social skills and practical skills.People with intellectual disability faced health risks from biological and environmental domains.Conclusion It is important to tailor to the unmet needs and health behaviors of people with intellectual disability. It is proposed to conduct capacity building for health professionals and institutions and improve the health of people with intellectual disability.
6.First trimester combined screening for Down's Syndrome with NT, Free-βhCG and PAPP-A
Zhengyou MIAO ; Tongkun SHI ; Yanjun GUO ; Qinhao SONG ; Huaxiang SHEN ; Ying XU
Chinese Journal of Laboratory Medicine 2012;35(10):932-935
Objective To explore the sensitivity of using NT,combined with serum biochemical markers (Free-βhCG,PAPP-A) for Down's Syndrome screening in early stage of pregnancy.Methods Collect pregnant women aged 17-45 years old voluntary antenatal screening in our hospital from March 2009to October 2010,a total of 11882 cases.Serum Free-βhCG and PAPP-A were measured NT value was determined by ultrasound at 11-13w+64 of gestation.Calculating combined screening (NT,Free-βhCG,PAPP-A),and serum integrated screening (Free-beta hCG,PAPP-A) risk,respectively,using the risk calculation software for the same person.Results Early pregnancy screening was performed in 11 882patients,18 had a fetus with Down's syndrome,a rate of 0.15%.The detection rates of Down's syndrome in combined screening and serum integrated screening were 83.3% and 72.2% respectively.The specificities were 98.4% and 97.3% and detection efficiency were 7.18%,3.90% respectively.Areas under the curve (AUCs) of fhst-trimester combined screening and serum integrated screening were 0.975 (95% CI:0.943,1.007),0.901 (95% CI:0.789,1.013) respectively.Conclusion In early stage of pregnancy,combined screening for Down's syndrome has higher sensitivity and specificity than serological screening,has higher detection rate in the same false-positive rate case,which can effectively reduce the pregnant women to receive invasive puncture.
7.Comparison of in vivo and in vitro Permeation Behaviors of the Ethosome Gels of Testosterone and Its Es-ters
Shu MENG ; Yang LI ; Cong ZHANG ; Jing QU ; Wei ZHANG ; Ying JIN ; Jing GUO ; Miao LI
China Pharmacy 2015;(22):3072-3074
OBJECTIVE:To compare in vivo and in vitro permeation behaviors of the ethosome gels of testosterone,testoster-one propionate and testosterone undecanoate. METHODS:The ethosome gels of testosterone,testosterone propionate and testoster-one undecanoate were prepared. With cumulative permeating amount and permeation rate as the indexes,Franz diffusion cell and HPLC were employed to compare in vitro permeation behaviors of 3 kinds of ethosome gels in mouse skin. With testosterone patch as the positive control drug, electrochemistry method was adopted to detect the concentration of testosterone in plasma 0,3,6, 9,12,24,36 and 48 h after applying such 3 kinds of ethosome gels on the back of rats,and then pharmacokinetic parameters were calculated with DAS 2.0 software. RESULTS:24 h cumulative permeating amounts of the ethosome gels of testosterone,tes-tosterone propionate and testosterone undecanoate were(234.31±13.8),(175.63±41.1)and(72.60±15.3)μg/cm2,and the per-meation rates were(10.25±1.9),(7.64±1.4)and(2.96±0.8)μg/(cm2·h),respectively. The pharmacokinetic parameters of the above-mentioned three kinds of ethosome gels and the positive control drug were respectively as follows as cmax of(20.19±2.57), (17.50±2.91),(0.23±0.04),(14.97±2.12)ng/ml,t1/2Ka of(2.80±0.45),(3.36±0.59),(4.02±0.62),(4.20±0.71)h,AUC0-48 h of(13.85±1.96),(13.93±2.13),(0.35±0.07),(11.76±2.31)ng·h/ml. CONCLUSIONS:in vivo and in vitro permeation behav-iors of the ethosome gels of testosterone and testosterone propionate are fairly good.
8.Multiple of the median correction of Down's syndrome serum markers with pregnant body weight
Zhengyou MIAO ; Yanjun GUO ; Tongkun SHI ; Qinhao SONG ; Xia LIU ; Ying XU
Chinese Journal of Laboratory Medicine 2014;37(1):46-49
Objective With Pregnant women weight correction for serum marker Multiple of Median (MoM) of First trimester and second-trimester,integrated screen-ing for Down's syndrome (DS),can reduce the false positive rate.Methods The same pregnant woman were taken venous blood vessels with sterile vacuum during the first trimester (11-13 W(+ 6) d) and the second trimester (15-20 W(+ 6) d),Alpha-fetoprotein (AFP),serum free beta-human chorionic gonadotrophin (Free beta hCG) and pregnancyassociated protein-A (PAPP-A) of three kinds of serum marker screening indicators were assayed by Using Time-resolved fluoroimmunoassay (TRFIA).Screening for risk assessment software was used to calculate serum marker Multiple of Median,To assess the risks of 7 997 cases of local pregnant women DS,To construct the weight equation of local population using nonlinear weighted regression method,With maternal weight correction for serum marker Multiple of Median (MoM) of local pregnant women,Comparing the changes of screening index MoM before and after correction,chi square test to compare the detection rate and false positive rate.Results MoM values of three kinds of serum markers (al-pha-fetoprotein,free beta subunit of human chorionic gonadotropin,pregnan-cy-associated plasma protein A) decreased with the weight increasing,Screening index MoM after correction weight equation,the screening of false positives for crowd from 4.12% down to 3.86% (x2 =0.021,P > 0.05).Setting threshold (cut-off) at 1/270,and no change detection rates were 71.4% the local population before and after correction weight equation.Conclusion Maternal weight may affect the results of Down's syn-drome sereening.When screening proposal to set up,it is worth making weight cor-rections for serum maker multiple of median in order to get accurate risk calculation results.
9.Experimental study of dual promoter baculovirus-mediated tumor-targeting radioiodine therapy
Min ZHANG ; Rui GUO ; Shuo SHI ; Ying MIAO ; Haoping XUN ; Biao LI
Chinese Journal of Nuclear Medicine and Molecular Imaging 2014;34(6):484-489
Objective To construct a recombinant baculovirus dual expression vector containing NIS gene under the control of human telomerase reverse transcriptase (hTERT) promoter and plasminogen kringle 5 (K5) gene driven by early growth response 1 (Egr1) promoter,and to explore the feasibility of targeting both tumor and tumor vessel with combination of radioiodide and antiangiogenic therapy.Methods The hTERT-NIS gene and Egr1-K5 gene fragments were subcloned into baculovirus vector,then packaged and amplified in the sf9 cells to obtain recombinant baculovirus Bac-hTERT-NIS-Egr1-K5.Bac-CMV-NISEgr1-K5,Bac-hTERT-0-Egr1-K5 and Bac-hTERT-NIS-Egr1-0 were constructed as controls.The expression of NIS and K5 genes in human cervix cancers cells (HeLa) was examined by Western blot and quantitative real-time PCR.Functional NIS activity was confirmed by the uptake of 125I,the inhibition of NaClO4 and the cytotoxicity of 131I.The apoptotic effect of 131I-inducedK5 on human umbilical veins endothelial cells (HUVEC)was analyzed by an apoptosis assay using flow cytometry.Statistical analysis was performed using the analysis of variance.Results The recombinant baculovirus Bac-hTERT-NIS-Egr1-K5 was successfully constructed.The NIS gene under the control of hTERT promoter was specifically expressed in HeLa cells.The baculovirusinfected HeLa cells showed a significant increase of 125I uptake,which was significantly inhibited by NaClO4(F199.296,P<0.05).Furthermore,a notable decreased cell survival rate (38.3%) was found after 131I treatment.The expression of K5 gene induced by 131I was elevated in a dose or time dependent manner and resulted in obvious inhibition with cell survival rate of 30.8% in baculovirus-infected HUVEC cells,which was significantly higher than that in the control groups (11.2% and 10.9% respectively,F=19.926,45.409;both P<0.05).Conclusions A recombinant baculovirus dual expression vector containing the NIS and K5 genes has been successfully constructed.This study suggests the feasibility of a synergistic strategy of NISbased raidoiodide therapy and K5-based antiangiogenic therapy in vitro,and make it possible to perform in vivo study in the near future.
10.Restaurant emissions removal by a biofilter with immobilized bacteria.
Jian-Yu MIAO ; Lian-Ying ZHENG ; Xiao-Fen GUO
Journal of Zhejiang University. Science. B 2005;6(5):433-437
Pseudomonas sp. ZD8 isolated from contaminated soil was immobilized with platane wood chips to produce packing materials for a novel biofilter system utilized to control restaurant emissions. The effects of operational parameters including retention time, temperature, and inlet gas concentration on the removal efficiency and elimination capacity were evaluated. Criteria necessary for a scale-up design of the biofilter was established. High and satisfactory level of rapeseed oil smoke removal efficiency was maintained during operation and the optimal retention time was found to be 18 s corresponding to smoke removal efficiency greater than 97%. The optimal inlet rapeseed oil smoke loading was 120 mg/(m(3) x h) at the upper end of the linear correlation between inlet loading and elimination capacity.
Air Pollutants
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isolation & purification
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Biodegradation, Environmental
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Cells, Immobilized
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Filtration
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instrumentation
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methods
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Microscopy, Electron, Scanning
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Oils
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metabolism
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Pseudomonas
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metabolism
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ultrastructure
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Restaurants
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Smoke
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Temperature
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Time Factors
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Waste Management
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instrumentation
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methods