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Objective To establish an evaluation system about animals infected with hantavirus,an observation of the BALB/c mice infected with hantavirus was made.Methods BALB/c mice were infected with hantavirus by intramuscular injection with stock solution.The specific antigen from BALB/c mice tissues after 3 days was detected with enzyme-linked immunosorbent assay (ELISA) and viral RNA with real-time polymerase chain reaction (RT-PCR).Results Within a short term,the specific antigen and viral RNA were detected from the brain and liver at day 3 after infection,but not be detected from the heart,spleen,lung,and kidney samples.Conclusions The results provided ones with some information on animals infected with hantavirus.
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Objective To develop a database management system specially applied for medical equipment maintenance administration, Methods Under PowerBuiider/SQL Server development environment, the C/S mode system structure was confirmed and the system function modules were designed. Results The application of C/S mode structure was coincident with the division-cooperation work flow in large and medium hospitals, and the design of function modules met the demand of information collection and utilization of medical equipment maintenance.Conclusion The database management system can preserve the information of medical equipment maintenance effectively and facilitate its usage. Moreover, many statistical indexes about the maintenance administration are available in this system.
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AIM: To analyze secondary structure and physico-chemical property of the fusion protein with bioinformatics network resource, and explore the expression of a secretory anti-osteoblastic carcinoma single-chain bi-functional antibody gene. METHODS: ①The single-chain variable fragment (ScFv) antibody gene and interleukin-2 (IL-2) gene were subcloned into corresponding restriction sites of retrovirus expression vector PLxSN. Mediated by liposome, the recombinant plasmid pL(ScFv-IL-2)SN was packaged with PA317 and selected in G418 to obtain the positive clones, which were able to produce stable retrovirus, and then osteosarcoma (OSC) cells were infected by the recombinant retrovirus, terming OSC/ScFv-IL-2. The virus titer was detected by using NIH3T3.②The transfected OSC9901 cells by ScFv-IL-2 gene were identified by polymerase chain reaction (PCR), reversed transcription-PCR and Western blotting. After the fusion protein was constructed, DNAssist and ANTHEPROT V5 softwares were used to analyze the amino acid sequence, the secondary structure, and the physico-chemical property of fusion protein. RESULTS: ①After the restriction enzyme and PCR identification, the pL(ScFv-IL-2)SN as a fusion protein expression vector, was constructed successfully, and high titer C26 cells were obtained; the expression of recombinant protein was confirmed by Western blotting.②On the fusion genes, the DNA sequence was analyzed with DNAssist nucleic acid sequence analysis software, and their secondary structure and physico-chemical property were analyzed with ANTHEPROT V5. CONCLUSION: The property of fusion protein can be analyzed and forecasted by means of bioinformatics network resources, and the approach may provide evidences for investigating single-chain bi-functional antibody gene.
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The signification and status quo of value engineering are introduced. Its application and significance in clinical medical engineering especially in the quality-control of medical instrument are also discussed.
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<p><b>OBJECTIVES</b>To identify differences in gene expression in renal and visceral adipose tissue in type 2 diabetic rats using cDNA representational difference analysis (RDA) and to explore the molecular pathogenesis of type 2 diabetes and its chronic vascular complications.</p><p><b>METHODS</b>A rat model of type 2 diabetes was generated by administration of a high fat and calorie diet combined with a low dose of streptozocin (STZ) injected into the tail vein. The difference bands were generated by cDNA representational difference analysis (cDNA RDA). The final difference products were ligated into the pUC-18 vector and sequenced. A bioformatics analysis was performed on the obtained expressed sequence tags (ESTs), and then the expression levels of known and novel genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). At the same time, full-length cDNA of a novel gene was cloned in silico.</p><p><b>RESULTS</b>The type 2 diabetic rats in this experiment experienced hyperglycemia, lipidemia, lower insulin sensitivity and normal body weight. We obtained 9 novel ESTs and 2 novel genes from renal tissue of rats and 6 novel ESTs and 1 known gene, the rat lipoprotein lipase (LPL) gene from their visceral adipose tissue. The 2 novel genes (RS91 and RS2) from the renal tissue were both very similar to serine (or cysteine) proteinase inhibitor, clade F and eukaryotic translation initiation factor 3 and subunit 5 (EIF-3 epsilon). The expression of both novel genes and the LPL gene were upregulated in renal and visceral adipose tissue of type 2 diabetic and fat-enriched rats. Full-length cDNA of the novel gene RS91 was cloned in silico.</p><p><b>CONCLUSIONS</b>(1) The rat model of type 2 diabetes generated in this study was ideal because the disease in the animals closely mimicked type 2 diabetic patients. (2) cDNA RDA is a flexible, inexpensive, more accurate, sensitive and highly effective technique for identifying differences in gene expression. (3) Six novel ESTs and 1 known gene were obtained from rat visceral adipose tissue. The LPL gene was upregulated in adipose tissue of type 2 diabetic and fat-enriched rats, a result possibly related to the diabetic animals' high fat and calorie diet, lipidemia, insulin resistance (RI) and molecular pathogenesis. (4) Nine novel ESTs and 2 novel genes were obtained from the renal tissue of rat. We believe the 2 novel genes to be the serine proteinase inhibitors clade F and EIF-3 epsilon in rats. The upregulation of the 2 novel genes in renal tissue of type 2 diabetic rats and may have been related to their renal impairment.</p>
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Animals , Male , Rats , Adipose Tissue , Metabolism , Cloning, Molecular , Diabetes Mellitus, Type 2 , Metabolism , Expressed Sequence Tags , Gene Expression Profiling , Kidney , Metabolism , Oligonucleotide Array Sequence Analysis , Plasmids , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Visceraالملخص
Ultrasonic estimation of fetal weight is important in the management o f labor and delivery. Lots of charts and formulas have been put forward in virtu e of regression methods. Artificial neural network, a kind of computer artificia l intelligence technology, can simulate human thinking based on neural structure and physiology. With the excellence in complex and non-linear information proce ssing, artificial neural network is fitter for the forecasting of fetal weight t han traditional regression methods.
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Objective To develop medical equipment management information system for medical engineering department. Methods Under PowerBuilder9.0/SQL and Server2000 development environment, the C/S mode system structure was adopted, and the system function modules, such as data input, browsing, querying, statistical analyzing, report generating, were designed. Results Data exchange between client and server was realized. Meanwhile, the designed function modules achieved normal operation. Conclusion The system structure and functions meet the requirements of working practice in medical engineering department.
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<p><b>OBJECTIVE</b>To purify recombinant nuclear protein of Hantaan virus.</p><p><b>METHODS</b>The recombinant plasmid was transformed into E.coli and induced by IPTG. The expressed protein is a fusion protein with GST and existed in inclusion bodies. The inclusion bodies were processed through denaturation and renaturation and were purified with Glutathione Sepharose 4B affinity chromatography column. Then the purified fusion protein and nuclear protein were examined by sandwich ELISA and Western blot.</p><p><b>RESULTS</b>The expressed fusion protein was separated from the mixture proteins by the first affinity chromatography. GST was cut from the purified fusion protein with thrombin. The nuclear protein was separated from GST by the second affinity chromatography. The crossed peak represents nuclear protein and the elute peak represents GST. The purified fusion protein and nuclear protein were single band by SDS-PAGE. Both of them had available antigen activity.</p><p><b>CONCLUSIONS</b>Purification of nuclear protein of Hantaan virus with Glutathione Sepharose 4B affinity chromatography is an effective method.</p>
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Blotting, Western , Chromatography, Affinity , Enzyme-Linked Immunosorbent Assay , Hantaan virus , Nuclear Proteins , Plasmids , Recombinant Fusion Proteins , Viral Proteinsالملخص
Objective To construct radiation inducible-specific hytk gene therapy vector using radiation inducible gene Egr-1 promoter,and to determine its feasibility in bladder carcinoma gene therapy. Methods A radiation inducible-specific hygromycin phosphotransferase-thymidine kinase fusion gene(hytk)vector was constructed by subcloning hytk gene into pCIneo/Egr-1 vector, which was carrying Egr-1 promoter.Then the vector was transferred into bladder carcinoma cells lines EJ with FuGENE TM 6.Antitumor effects were observed after irradiation with 60 Co-rays as well as gancyclovir(GCV) treatment. Results The new cell line,EJ/Egr-1-hytk,incorporated and expressed the hytk gene,which was proved by RT-PCR and Western blotting.Egr-1 promoter actively drived hytk gene expression in the presence of ?-rays in the EJ cells, while lower hytk expression was detected without ?-rays.Antitumor effects were observed after ?-rays irradiation with 60 Co rays and GCV treatment. Conclusions Hytk-based vector harboring Egr-1 promoter is a novel ideal candidate vector for bladder carcinoma gene therapy.
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AIM: To investigate the effects of dexamethasone(DEX) on the pancreatic endocrine function when steroid diabetes mellitus(SDM) occurred. METHODS: 140 rats were randomly assigned to 4 groups according to the clinic doses of DEX administration, Radioimmunoassay and in situ hybridization were applied to evaluate islet hormone changes in pancreas and blood.RESULTS: Changes in fasting blood glucose(FBG) ?fasting serum insulin(FINS) caused by DEX were dose and time-dependent. Changes in somatostatin(SS)mRNA and protein in islet as well as FINS occurred earlier than that of FBG. CONCLUSION: The super-physiological DEX influenced the expression SS and insulin secretion in islet, which may play an important role in SDM.
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AIM: To facilitate the suicide gene delivery into neoplasm, a chimeric gene of HSV-tk and green fluorescent protein (gfp) was constructed. METHODS: Molecular cloning technique was used to construct this kind of eukaryotic vector. The internal ribosome entry site (IRES) of encephalomyocarditis virus (EMCV), which could coordinate expression of two genes in a single vector, was optioned. By using liposome-mediated transfection, eukaryotic expression vector tgCMV/hytk-IRES-gfp was transfected into human bladder carcinoma cells EJ. RESULTS: A bicistronic eukaryotic vector carrying gfp and hygromycin phosphotransferase-thymidine kinase fusion (hytk) gene was constructed. The results of PCR and microscopy detection show that the hytk-IRES-gfp gene was successfully transferred into EJ cells. There were no differences in the growth pattern or the morphology between EJ and EJ/hytk-GFP cells. In vitro experiments demonstrated dose- and time-dependent cell killing by transduction of the hytk-IRES-gfp gene followed by GCV treatment. The IC50 (the concentration required to elicit 50% growth inhibition) was 2.16 mg/L in treatment with GCV for 72 hours. CONCLUSION: These results suggest that this new kind of eukaryotic vector could serves as a new tool and method for neoplasm gene therapy.
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This paper introduces the applications of network technology, wire and wireless technologies to designs of medical equipments. The theories and applications of such technologies are presented as the ones of serial communication, concurrent communication, USB interface, Bluetooth HomeRF, wireless network communication and GPRS.
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HIFU treatment allows the destruction of a precise volume of tissue within an tumour without harming surrounding structures.The FEP-BY02 model is one of commercially available machines based on this technique,which is proved effectively in clinical practice.It's basic architecture,functional principle and some suggestions for safety use are introduced in this paper.
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Biomedical sensor is a key component to the modern medical instrument,which can convert physiological value to electrical signal.After introducing and analyzing the definition,main uses and present application of biomedical sensor,this paper lays emphasis on its developing trend.
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Abstract PCB faults on X-ray machine occur frequently in our hospital due to humid climate, which lead to many malfunctions such as potentiometer failure, blank screen, lacking KV display, unavailable parameter choosing and disappearing of Er1～Er7 fault code. These malfunctions could be diagnosed and repaired quickly based on analyzing the status of the machine's workflow indicator lights, as well as studying circuit and function principle of the equipment.
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Objective To investigate the difference of gene expression in the visceral adipose tissue of the lean type 2 diabetic rats. Methods The difference bands were generated by the representational difference analysis ( cDNA RDA) ; the final difference products was ligated into the pUC-18 vector, subcloned and sequenced, and the obtained expressed sequence tags ( ESTs) were given bioinformatics analysis, and then the expression level of known gene was verified by semi-quantitative RT-PCR preliminarily. Results Six novel ESTs and 1 known gene [ rat lipoprotein lipase ( LPL) gene) were obtained in visceral adipose tissue of rat. One of the ESTs was partially similar to mouse musculus ERA-like GTPase (Era) gene. LPL gene was up-regulated in the visceral adipose tissue of the lean type 2 diabetic rats and those fed with fat-enriched food. Conclusion Six novel ESTs and 1 known gene in high expression are obtained from visceral adipose tissue of rat. LPL gene among them is up-regulated and may be related to the high fat and high calorie diet, hyperlipidemia, insulin resistance and molecular pathogenesis of the lean type 2 diabetic rat.
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Using the technique of fluorescent-labled mRNA differential display, new apoptosis related gene 2ass-bnip3 of type 2 diabetic cardiomyopathy was found, the sequence of 1594 bp with coding 187 amino acids was obtained by the full-length clone, and its structural and functional predictions were performed. 2ass-bnip3 may play an important role in the development of diabetic cardiomyopathy via a regulatory pathway of calcium regulation and apoptosis.