Objectives To evaluate the diagnostic value of real-time quantitative fluorescence polymerase chain reaction( Q-RT-PCR ) assay and immunofluorescence assay for diagnosis of hMPV. Methods Totally 1 283 children with acute respiratory infection admitted in Jiaxing Maternity and Child Health Care Hospital for treatment from November 2008 to May 2009 were recruited in this study. The hMPV positive stains were separated and sequenced in this area. The sequences between the local hMPV stains and Holland stains NLD00-1 were compared. The specific primers and fluorescent probe were designed according to the sequence of epidemic hMPV strain. The Taqman methodology was applied in Q-RT-PCR. Negative pressure suction was used to acquire nasopharyngeal secretions specimens. Both Q-RT-PCR and immunofluorescence with FITC labeled monoclonal antibody were used to analyze them, respectively. The McNemar, test was applied to analyze the correlation between the two methods. Results Totally 1 283 specimens were analyzed with Q-RT-PCR and immunofiuorescence simultaneously. Q-RT-PCR analysis showed there were 59 cases positive. Immunofluorescence analysis showed there were 55 cases positive. Fifty-two cases were positive in both assays. There were 7 cases positive in Q-RT-PCR assay but negative in immunofluorescence assay and 3 cases negative in Q-RT-PCR assay but positive in immunofluorescence assay. If Q-RT-PCR method was set as the golden standard, the sensitivity and specificity for immunofluorescence detection method were 88. 1%and 99. 8%, respectively. Positive predictive value and negative predictive value were 94. 5% and 99. 4%,respectively. There was no significant difference ( χ2= 0. 9, P ＞ 0. 05 ) by McNemar' test between the two methods. Conclusion The diagnostic value of immunofluorescence assay is close to Q-RT-PCR assay.

Objective To explore the effect of prenatal inflammation on vascular remolding and blood pressure in mid-aged rats.Methods Time-mated pregnant Sprague-Dawley(SD)rats were randomly divided into 2 groups,received peritoneal injection of 0.79 mg/kg LPS on the gestation days 8,10,and 12,or same volume of the sterile saline at the same time points.Nine pups were randomly selected from each group for the later experiments,and the offspring were named as LPS groups for those having prenatal LPS exposure and control group for those having not.Their blood pressure was determined with a rat tail non-invasive instrument by tail-cuff method from 6 weeks old to 35 weeks old,and then their aortas were taken out for media thickness(MT),diameter of lumen(LD),and the ratios of MT/LD.The expression of proliferating cell nuclear antigen(PCNA)on the vessel was detected by ELISA.The serum level of NO and plasma endothelin-1(ET-1)were measured by nitrate reductase and radioimmunoassay respectively.Results Compared with control group,LPS groups had significantly raised blood pressure,a significantly higher ratio of MT/LD,obviously increased expression of PCNA,and markedly elevated serum ET-1.Conclusion The offspring whose prenatal rats were exposured to LPS result in vascular remolding,vasofunctional disturbances and hypertension.

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 ; Biosensing Techniques ; Electrodes ; Models, Theoretical ; Monte Carlo Method ; Nanoparticles ; Particle Size ; Platinum

Humans ; Work

Private Sector ; Public Sector ; Social Responsibility

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.

Human proximal tubule epithelial cell line,HK-2 cells,were cultured with various concentrations of insulin for 48 h.Human urate transporter (hUAT) mRNA was detected by realtime quantitative PCR.hUAT mRNA levels were down-regulated by insulin (5,25,125,500 μIU/ml)in a dose-dependent manner (relative expression median were 0.95,0.40,0.24,and 0.23).In vitro,the expression of hUAT mRNA in HK-2 cells is associated with the concentration of insulin.

Objective To investigate the soft contact lens related knowledge and wearing behaviors.Method Two hundred outpatients in the ophthalmologic clinic wearing SCL involved in the survey by the questionnaire to investigate the SCL-related knowledge and the wearing behaviors.Results In the 200 SCL wearer,73.50% did not make any examinations by specialists and only 18.00% of them took regular examinations.The wearers were good at the knowledge of selecting SCL diopter and positive and negative lens.The wearers with a history of less than 6 months were better than those wearing SCL more than 6 months in respect of hand cleaning,lens check,lens wearing, and lens cleaning.The wears with the education of two-year vocational training were significantly better than those with the education of three-year or more collegiate training in respect of wearing duration,sleep with lens,hand cleaning and lens cleaning(all P<0.05).Conclusion Contact lens wearers know less about SCL related knowledge and their wearing behaviors needs improvement.

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.

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.