Application of SPR protein chip in screening for imported malaria / 生物工程学报

Imported malaria has become a major risk factor for malaria prevention and control in China. How to screen malaria quickly for people entering China is an urgent problem to be solved. Protein microarrays are widely used in high-throughput screening and diagnosis. In this study, surface plasmon resonance (SPR) technique for malaria detection was established by using the specific adsorption surface treated by polyethylene glycol polymer, and the malaria specific antigen HRP2 was used as capture probe. The optimal concentration of antigen, sensitivity and specificity of detection, as well as anti-interference ability of the chip were analyzed. The SPR protein chip was applied to detect specific antibodies of malignant malaria in serum with the advantage of label-free, instant and fast. Compared with fluorescence quantitative PCR, there were no significant difference in sensitivity and specificity between the two methods. This study lays a foundation for further development of protein microarray for malaria typing identification, and it is conducive to the rapid screening of malaria for people entering.

Application of iPDMS protein microarray in screening of tumor-associated antigen autoantibodies / 生物工程学报

The rapid screening of tumor markers is a challenging task for early diagnosis of cancer. This study aims to use highly sensitive chemiluminescent protein microarray technology to efficiently screen a variety of low abundance tumor related markers. A new material, termed integrated polydimethylsiloxane modified silica gel (iPDMS), was obtained by adding a surface polymerization initiator with olefin end to the conventional polydimethylsiloxane, and fixing into the three-dimensional structure of polydimethylsiloxane by thermal crosslinking through silicon hydrogen bonding. In order to make the iPDMS material resistant to non-specific protein adsorption, a poly(OEGMA) polymer brush was synthesized by surface-initiated atom transfer radical polymerization at the active initiation site. Finally, 20 tumor-related antigens were printed into the specific areas of the microarray by high-throughput spray printing technology, and assembled into 48-well detection microtiterplates of the iPDMS microarray. It was found the VEGFR and VEGF121 autoantibodies that obtained from 8 common tumors (breast cancer, lung cancer, colon cancer, gastric cancer, liver cancer, leukemia, lymphoma and ovarian cancer) can be used as potential tumor markers. The chemiluminescence labeled iPDMS protein microarray can be used for the screening of tumor autoantibodies at early stage.

Clinical value of magnetic resonance diffusion-weighted imaging in early diagnosis and treatment of cervical lymph node recurrence after radiotherapy of nasopharyngeal carcinoma / 中华放射肿瘤学杂志

Objective To explore the clinical value of magnetic resonance diffusion-weighted imaging ( MR-DWI ) in the early diagnosis of cervical lymph node recurrence after radiotherapy of nasopharyngeal carcinoma, aiming to provide reference for targeted diagnosis and treatment of these patients. Methods The MR-DWI features of 17 patients with recurrent cervical lymph nodes after radiotherapy from 2005 to 2016 were retrospectively analyzed. The results of diagnosis and treatment after lymph node recurrence were summarized. Results The recurrent lymph nodes of 17 patients showed a high signal or mixed signal on MR-DWI images. The sensitivity of MR-DWI and T2WI fat suppression sequence was 100% and 60%. Positron emission tomography-computed tomography ( PET-CT) or biopsy was performed to validate the diagnosis in patients with highly suspected single cervical recurrence. Besides, surgical treatment yielded better clinical prognosis. Conclusions MR-DWI is highly sensitive to recurrent cervical lymph nodes of nasopharyngeal carcinoma after radiotherapy, especially for the small lymph nodes of 5-10 mm in diameter, which are easily ignored. PET-CT examination should be performed, the nature of the lymph nodes should be confirmed by multi-modality imaging diagnosis, and timely operation has important clinical significance in improving the therapeutic effect and quality of life for patients with cervical lymphnode recurrence.

The application of SPR technology in prenatal testing of fetal RHD blood type gene / 国际检验医学杂志

Objective To study the feasibility of detecting fetus RhD type gene by Surface Plasmon Reso-nance(SPR)technology,and to establish a new rapid diagnosis method for fetus RhD type gene.Methods The different types of DNA corresponding RNA probes were fixed on the surface of SPR chip by using amino cou-pling methods,and optimize the chip analysis condition,and then using the RNase H enzyme hydrolysis,signal amplification detection,at last the detection conditions were determined.We use the RhD type gene exon 5,7 of RNA probe to test its corresponding DNA molecules,and analyse the specificity and sensitivity of SPR chip detection signal.Results The SPR technique for detecting the exon 5,7 of RhD blood type gene shows good sensitivity and specificity in all,SPR technology can specifically detect the Exon 5,7 of RhD blood type gene, and the sensitivity of for detecting RhD gene exon 5,7 is 100 pmol/L by SPR.Conclusion The SPR technolo-gy can quickly detect RhD gene accordingly,SPR technology is simple,rapid,reliable and label-free,w hich can provide a new way predicting fetal RhD type for RhD negative prenatal.

Imaging diagnosis of the medial group of retropharyngeal lymph nodes and the exploration of the optimization of target volume for protecting the swallowing structure in nasopharyngeal carcinoma / 中华放射肿瘤学杂志

Objective To clarify the diagnostic value of diffusion-weighed imaging ( DWI) in the medial group of retropharyngeal lymph nodes in nasopharyngeal carcinoma, understand the clinical characteristics of retropharyngeal lymph nodes and explore the feasibility of optimizing the target volume of CT V60. Methods Clinical data of 437 patients with clinical stage Ⅰ-IVa nasopharyngeal carcinoma admitted to Jiangsu Cancer Hospital from 2011 to 2017 were retrospectively analyzed. All patients underwent magnetic resonance imaging (MRI),DWI (1 000 s/mm2) and enhanced CT scans to analyze the clinical characteristics of retropharyngeal lymph nodes and investigate the dosimetric advantage and safety of CT V60 lower margin on the upper margin of C2. Results The medial lymph nodes with a transverse diameter of 2. 0-19. 0 mm were detected 13 of 437 patients,and 53. 8% of the lymph nodes were measured 2-5 mm in transverse diameter. The medial lymph nodes were distributed between the superior margin of C1and 1/3 of C3.Its occurrence was related to N stage,double cervical lymph node metastases,especially the transverse diameter of cervical lymph node＞ 3 cm.The sensitivity of DWI,T2and enhanced CT were 100%,61. 5% and 23. 1%.After the special cases were excluded,the lower margin of CT V60on the superior margin of C2was separated. The radiation dose and volume of the swallowing structures were significantly decreased. The 5-year survival rate was 80% without recurrence in the optimized region. Conclusions The incidence of the medial group of retropharyngeal lymph nodes is low with a diameter of less than 5 mm. DWI possesses advantages in displaying the medial group of retropharyngeal lymph nodes. Isolating the lower margin of CT V60from the superior margin of C2is safe and feasible and has dosimetric advantages for protecting swallowing structure.

Study on high-throughput respiratory tract virus detection method based SPR / 国际检验医学杂志

Objective To develop a biosensor method with strong specificity and high‐throughput by combining with the surface plasmon resonance (SPR) and gene chip technique and aiming at 9 kinds of common respiratory tract viruses including influenza A and influenza B ,(Influ A ,B) ,H1N1 ,respiratory syncytial virus (RSV) ,parainfluenza virus 1 -3 (PIV1 -3) ,adenovirus (ADV) and coronavirus (SARS) leading to severe acute respiratory syndrome .Methods Firstly the software primer 5 was used to design the specific primer and probe of related viruses in the conserved sequence ;the designed nine kinds of corresponding respiratory virus probes were immobilized in the specific region of SPR chip after chemical modification .The SPR technique was applied to conduct the real time monitoring the hybridization process of the probe with the PCR products .Finally the signal amplification was realized by the biotin and streptavidin system .Results The designed gene chip could detect 9 kinds of respiratory tract viruses by high‐throughput with better detection specificity ;the chip surface could be reutilized after certain regeneration condition ,which avoided the influence of intra‐batch difference on the results ;the detection sensitivity reached the nanomole level .Conclusion The prelimi‐nary study results demonstrate that using the SPR biosensor technique to establish a high‐‐throughput detection of respiratory tract viruses has some practicability and feasibility ,and is expected to become a rapid ,large scale and high‐ throughput measure for screening respiratory tract viruses with good application prospect .

Surgical treatment of pancreatic sinistral portal hypertension and literature review / 中华肝胆外科杂志

Pancreatic sinistral portal hypertension is a localized kind of portal hypertension that usually occurs as a result of the splenic vein obstruction caused by pancreatic diseases.Furthermore,it is also an important cause of upper gastrointestinal hemorrhage.Management in clinical practice should be directed at the sinistral portal hypertension and primary pancreatic diseases.

Detection of platelet antibody by surface plasmon resonance technique and its preliminary application / 重庆医学

Objective To study the platelet antibody screening and crossing match by surface plasmon resonance(SPR) ,and to find a new way for platelet compatibility testing .Methods The corresponding universal platelet antigen was fixed on the SPR chip surface by the amino coupling method .Platelet antibody positive and negative control serum were analysed by SPR micro-ar-ray ,the stability ,sensitivity and specificity of this technique were discussed ,and compared with MAIPA assay .Finally we used the SPR technology to cross match for ten cases of the platelet antibody positive patients before infusion ,and to evaluate the effect of platelet infusion .Results For the SPR technology ,the stability ,sensitivity and specificity of platelet antibody detection were all better ,106 cases of the repeated platelet transfusion samples were tested by SPR assay and MAIPA method ,there was no signifi-cant difference between them(chi-square=0 .333 ,P>0 .05) ,the total consistency was 97 .2% .The 10 cases of platelet antibody positive patients were crossed match before platelet transfusion by SPR technology ,the good results of 8 cases of them were found by the clinical tracking evaluation ,1 h CCI>7 .5 ,24 h CCI>4 .5 .Conclusion SPR technology for screening platelet antibody mat-ches with MAIPA method in basic quality ,but SPR assay is simple ,rapid ,reliable and intuitive ,label free ,which can satisfy the re-quirements for clinical rapid detection of platelet antibody screening and crossing match .

Effects of alprostadil in the prevention of portal vein thrombosis after splenectomy and devascularization: a historical control study / 中华肝胆外科杂志

Objective To investigate the clinical outcomes of alprostadil in prevention of portal vein thrombosis after splenectomy and devascularization.Methods 113 patients with PHT who were treated with prophylactic alprostadil after splenectomy and devascularization procedures from May 2009 to Apr 2013 were included into the treatment group.112 conservative patients with PHT who were treated with traditional prophylactic anticoagulants after the same operations before May 2009 were included as the control group.The postoperative complication rates,mortality,postoperative drainage volume from the abdominal cavity,blood platelet counts,prothrombin time,liver function,Child-Pugh's scores and portal vein thrombosis rates between the two groups were compared.Results When compared with the control group,the postoperative complication rate and mortality in the alprostadil group were not increased,while the postoperative drainage volume from the abdominal cavity was significantly reduced.The increase in blood platelet counts and prothrombin time were similar in the 2 groups.Furthermore,the extent of hepatic dysfunction on the 3rd and 7th after operation was significantly decreased.On short term follow-up,color droppler ultrasonography showed the portal vein thrombosis rate of the treatment group was significantly lower than the control group,with less extensive degree of thrombosis in the treatment group.Conclusion Alprostadil is a safe and effective anticoagulant which provided better prevention of portal vein thrombosis after splenectomy combined with devascularization.

Preparation and performance assessment of Gamma-peptide nucleic acid gene chip detection system based on surface plasmon resonance / 生物医学工程学杂志

The aim of this study was to build a gene chip system with surface plasmon resonance (SPR) technique, for which Gamma-peptide nucleic acid (Gamma-PNA) functioned as a probe, in order to improve sensitivity and its specificity. With the use of self-assembled monolayer (SAM) technology, surface chemistry of two-dimensional structure was used. Gamma-PNA was designed according to the bioinformatics, and was plated on the SPR chip modified by SAM. Subsequently, relevant parameters of the experiment were ensured and optimized. The results showed that the performances of Gamma-PNA probe was little affected by the ion concentration of buffer, and it had a strong light signal in a stable state. As the ion concentration was 0, there were still good hybrid reactions; pH value had less influence upon Gamma-PNA probe, and acid environment of buffer could be better. Gamma-PNA probe combined with sensor technologies achieved made the probe with dispensable labels and real-time detection. It also improved the efficiency of the hybridization and the stability, providing the foundation for clinical application.

Study on morphology of surface atoms conformation of nanogold-based genechip / 生物医学工程学杂志

Conformations of surface atoms in various stages of nanogold-based genechip testing were scanned by the atomic force microscope based on the scanning tunneling microscope. The findings were: First, the surface atoms of genechip slide (formylphenyl glass) were in a regular porous-arrangement; Second, after combination with probe, the regular porous arrangement changed to be irregular; Third, after hybridization with the target nucleic acid, the surface atoms were once again in a cable-like arrangement which was relatively structured and intensively cross-parallel. However, after the silver staining, the surface atoms showed a larger block structure with serious unevenness. From these results we can intuitively know the process and differences in probe combination, nucleic acid hybridization, and silver staining. Moreover, the relevant experiment was verified at the micro-level.

The preparation and testing analysis basis of gene chip checking system with surface plasmon resonance imaging / 生物医学工程学杂志

The detection method of gene chip based on SPR principle is a potential high-throughput microanalysis method without labelling. With the use of Self-assembled monolayer (SAM) technology, the gene chip of Neisseria gonorrhoeae probe lattice has been prepared, detected and analyzed using the Surface plasmon resonance (SPR) and SPR imaging (SPRI) gene chip detection system here-in provided for research in the hybridizatin reaction on the probe lattice of gene chip. The result indicates that there is an obvious resonance assimilate peak on the SPR resonance curve. And after hybridization, the refractive index and resonance as well as molecular weight of the probe have increased. So whether a hybridization takes place or whether the wanted ingredient is in the sample under examination can be determined by using SPR to watch the detecting interface or the resonance curve. The SPRI detection system is available for observing the happening of a hybridization on the probe of gene chip in real-time and straighforwardly. The SPR and SPRI system can do analysis qualitatively and quantitatively.

A study on surface plasmon resonance-based gene chip / 生物医学工程学杂志

The surface plasmon resonance (SPR)-based gene chip was prepared according to the following processes: First, a film of nanogold, which was synthesized by using Frens' method, was plated on chip by Chlorauric acid/hydroxylamine method. Then probes were fixed on nanogold film by Self-assembled monolayer (SAM) technology. Subsequently, the fixing time and concentration of probes, the sensitivity and the specificity of the chip were optimized. Our results suggested that the chip plated with 2.5 nm nanogold film has a better SPR reflection, and when fixed by probes for 4.5 h at the concentration of 1 500 nmol/L, the gene chip also shows a fine performance of detection and can identify accurately the mismatch between bases in SPR detection system. The gene chip constructed in the research can be used for SPR sensor detection.

A study on surface plasmon resonance-based gene chip system for rapid pathogen detection / 中华检验医学杂志

Objective To study application of surface plasmon resonance(SIR)system in detection of clinical pathogen with a gene chip.Methods 27 clinical samples were detected by SPR-based gene chip system.These samples were composed by 8 positive blood samples,3 positive pyoid samples,9 positive leucorrhea samples and positive reproductive tract pyoid samples,1 positive biopsy sample and 6 negative biopsy samples.Specific primers and probes for target pathogens were designed by bioinformatics methods and validated by PCR and enzyme-labelled chemiluminescence,respectively.SPR-based gene chip was prepared and utilized to detect clinical samples by SPR system.Results The primers and probes showed good specificity and accuracy,which can be applied to perform PCR and application of the gene chip.Compared with the clinical analysis,gene chip analysis of 26 clinical samples showed the consistent results.Conclusions SPR detection system proved to be accurate and reliable.The chip will have a promising prospect in application.

Nanogold-based Gene Chip for Rapid Pathogen Detection / 中华医院感染学杂志

OBJECTIVE A practical gene chip which aimed to detect and identify pathogens rapidly and exactly is developed on the basis of patent technology of nano-enlargement-detection. METHODS Oligonucleotide probes for the specific gene fragments of target pathogens were designed and immobilized on gene chip.Target sequences were labeled by nanogold as reporter materials.After hybridization,its results were recorded by the interaction between nanogold and silver which amplified the hybridization signal to form brown particles,which could be detected by naked eyes. RESULTS The probes designed were all of strong specificity and great reliability possessing identity of hybridization conditions.The reaction time for marking could be decreased by properly raising the ratio of nanogold and nucleic acid and the speed of labeling reaction could be fastened significantly by gentle agitation.A better hybridization results could be obtained when the samples were hybridized for 8 hours at 45℃ with 0.8 mol/L ionic strength,and then strictly rinsed.Furthermore,the hybridization efficiency could be increased remarkably by slight circumgyratation.A better chromatic effect resulted from the reaction way in 3min?3 at 37℃.The sensitivity of gene chip assays in this test could reach to 100 fmol/L.Compared with traditional detection approach,detection by the chip displayed such advantages as speediness and simplicity and the detection results could be easily recognized by naked eyes. CONCLUSIONS The chip detection technology has met the demand of design exhibiting high sensitivity,strong specificity,and easy operation without special device and showing a promising prospect.

Coagulase-negative Staphylococci and ica Operon Examination:A Clinical Study / 中华医院感染学杂志

OBJECTIVE To investigate the significance of CNS in clinical infections.METHODS A total of 114 CNS strains isolated from our hospital were identified by conventional procedures and the icaD gene was amplified by PCR.RESULTS Of all CNS strains,the highest isolated rate was S.epidermidis(41.2%).CNS isolated from deep venous catheters,wound secretions and blood had a higher rate of carrying ica operon,accounted for 44.4%,42.1% and 36.8%,respectively,whereas 24.0% in respiratory secretions and 14.1% in urine.Among the ica operon positive CNS strains,the percentages of S.epidermidis and S.haemolyticus were 42.6% and 19.0%.CONCLUSIONS There is a wide range of CNS species carrying ica operon,especially in S.epidermidis.CNS isolated from different specimens might have different significances.It should be cautious to assess the results of isolated CNS from respiratory and urine specimens.The CNS isolates from blood specimens might be contaminated.The PCR method for the ica operon is simple and easy.

Preliminary clinical experience of laparoscopic repair of esophageal hiatal hernia / 中国微创外科杂志

Objective To summarize the clinical experience of laparoscopic repair of esophageal hiatal hernia.Methods A total of 15 cases of esophageal hiatal hernia underwent laparoscopic hernia repair and fundoplication from May 2004 to April 2005 in this division.There were 4 cases of type Ⅰ hernia(all of which presented severe gastroesophageal reflux disease),10 cases of type Ⅱ hernia,and 1 case of type Ⅲ.Surgical procedures included laparoscopic Nissen total fundoplication in 9 cases,Toupet partial fundoplicatin in 4 cases,and Dor partial fundoplication in 2.Symptoms of gastroesophageal reflux disease,including heart burn,dysphagia,regurgitation,chest pain,and belching,were evaluated by using the Visual Analogue Scales(VAS) at preoperative period,1 postoperative month,and 6 postoperative months,respectively.Results No conversion to open surgery was required in this study.The operative time ranged 100~187 min(mean,125 min).The postoperative hospital stay was 2~5 days(mean,2.8 days).All the patients were followed for 1~12 months(mean,8.5 months).No hernia recurrence was found.The VAS scores decreased from 5.0?3.9 preoperatively to 0.9?1.3 at 1 month postoperatively(t=3.823,P

Detection of Bacterial Pathogens by Nanogold-based Gene Chip Combining with One-time PCR with Common Primers / 中华医院感染学杂志

OBJECTIVE To develop a preparation technique of sample of one-time PCR with common primers based on ribotyping which was combined with the detection system of nanogold-based gene chip to detect clinical bacterial pathogens.METHODS According to the highly conserved regions of rDNA,the common primers were designed and used to amplify each target bacterial ISRs by one-time PCR,and the specific oligonucleotide probes for each target ISRs were designed,utilized to establish the new nanogold-based gene chips.After the characteristics of the chip such as sensitivity,specificity and reliability were determined,the chip was used to detect clinical samples.RESULTS The designed common primers could amplify the 12 target bacteria successfully by one-time PCR.All selected probes were of strong specificity and great reliability.The chip had high sensitivity,specificity and reliability,reaching 50 fmol/L of detection sensibility.Clinical detection results showed the chip had a great accuracy.CONCLUSIONS Compared to multi-PCR chip detection,the detection procedure and complexity of the chip are decreased significantly,and have more practical value in clinical pathogens detection.

Simultaneous detection of multiple pathogens by nano-gold-based gene chip combined with restrictive enzyme digestion without PCR / 解放军医学杂志

Objective For realizing the simultaneous detection of multiple pathogens,by looking for a method with a combination of the new gene chip detection system based on nano-gold with the technology of restriction endonuclease without PCR.Methods Helicobacter pylori,Mycoplasma pneumoniae,Chlamydia trachomatis,Candida albicans,Ureaplasma urealyticum,and EB virus were selected as the experimental targets.Endonuclease Hha Ⅰ was selected as tool enzyme.After bering digested by Hha Ⅰ,the digested fragments of samples were tailed with poly-A.The samples were then detected by the gene chip detection system based on nano-gold.Both specific and common probes were used in the hybridization.The coincidence rate of the detection results between the new constructed chip test and the fluorescence quantitative PCR test in 168 clinical samples was examined.The stability and sensitivity of chips detection were also checked.Results The new constructed nano-gold-baesd gene chip combined with restrictive enzyme digestion without PCR could be used to detect the target pathogens.The coincidence rate of the chip detection test and fluorescence quantitative PCR test in 168 clinical samples was 89.2%.Chip detection results showed that the stability of chips detection was 100% and the sensitivity was 50pmol/L.Conclusion The newly constructed nano-gold-baesd gene chip combined with restrictive enzyme digestion without PCR can be widely applied in the simultaneous detection of Mycoplasma,Chlamydia,fungus,virus and bacteria.It shows a bright prospect in increasing the throughput of identifieation of pathogene.