Expression of NDV HN protein in rice and development of a semi-quantitative rapid method for detection of antibodies / 生物工程学报

The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log2, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.

Development of Colloidal Gold Detector Instrument Based on CMOS Image Processing / 中国医疗器械杂志

This paper introduces a kind of immune colloidal gold detector instrument from the aspects of machinery,hardware and software.The instrument first collects one image through a CMOS sensor and then analyzes the image with image processing algorithm on Linux platform.Firstly,the instrument sets and stores the parameters separately for each test item,and then calls the saved item parameters when testing the item sample.So,the instrument can be used in a variety of fields and items.In this paper,a quantitative experimental test on C-reactive protein sample was performed,and the results indicate the coefficient of determination what denoted equal to 0.99,and the repeatability is greater than 93%.

Development of Semi-quantitative Solid Sampling Mercury Analyzer for Cosmetics / 分析化学

On the basis of the chromogenic reaction between Hg and CuI, a semi-quantitative solid sampling Hg analyzer comprising the catalytic furance, Hg testing tube, air pump and smart cellphone was developed. White carrier 101 was chosen as the adsorbent for CuI to react with Hg from the catalytic furnace. The established Hg analyzer can not only visually recognize the coloration when Hg exceeding the limit standard, but also semi-quantitatively detect the Hg content in cosmetics fast using a smart cellphone and RGB analysis software, after direct solid sampling introduction of cosmetics sample. The instrumental detection limit ( LOD) of mercury was 50 ng, the linearity ranged from 50 ng to 2500 ng, the linear regression coefficient ( R2) was higher than 0. 97, and the RSD of the corresponding RGB values was 6% ( n=11 ) . Nine real cosmetics samples were measured by the established method, whose relative differences of Hg contents with that by the standard method (Safety Technical Specification for Cosmetics, 2015 edition) were less than 10% . The whole analytical time can be controlled within 5 min. The established instrumental method is simple, fast, accurate and visual, and extremely suitable to fast and on-site monitoring of Hg in cosmetics samples.

Evaluation and development of preoperative liver reserve function evaluation methods / 中华消化外科杂志

Liver resection is one of the important treatments of liver diseases,especially hepatocellular carcinoma.In China,the vast majority of liver cancer patients suffer from generalized damage of the liver parenchyma such as cirrhosis,lead to liver reserve function reducing in various degrees.Liver dysfunction or even liver failure after liver resection becomes an important reason of perioperative death and influences the patients' long-term survival.Therefore,accurate preoperative evaluation of liver reserve function is very important.Though there are seveal kinds of assessment of liver reserve function in recent years,it still lack of a clinically recognized,comprehensive assessment method.This paper reviewed the clinical commonly used preoperative liver reserve function evaluation methods,summarizes and analyzes the value and the insufficiency of several important methods,and prospects the development of evaluation methods about preoperative liver reserve function.

CMV Disease / 대한이식학회지

Cytomegalovirus, one of beta human herpes virus, cause significant morbidity and mortality in the renal transplant patients via direct and indirect effects of viral infection, which is defined as the presence of CMV in the host. CMV disease in renal transplant recipients is the result of direct cytopathic effects of proliferating virus and manifest as CMV syndrome and/or tissue-invasive disease. Indirect effects of CMV infection are caused by the long-term immuological responses of host to existing CMV virus and manifest as increased occurrence of acute rejection, predisposition to opportunistic bacterial, viral or fungal infection, development of lymphoproliferative disease, atheroscrelosis and posttransplantation diabets mellitus and increase in all-cause mortality in renal transplant recipients. Because the development and severity of CMV disease depends on the amount of virus present in renal transplant recipients, the quantitative tests for viral load such as antigenemia assay and molecular amplification of CMV are useful for diagnosis of CMV disease and serve as a guidance in prophylaxis and treatment of CMV disease in renal transplant recipients. CMV prophylaxis is indicated in high-risk patients such as CMV negative recipients of CMV positive donor or CMV positive recipients who have received anti-lymphocyte antibodies as induction therapy or treatment of acute rejection. Universal prophylaxis and pre-emptive treatment using mainly ganciclovir are main strategies and have their own advantages and disadvantages. Recently long-term prophylaxis up to 24 weeks is favored to prevent the occurrence of CMV disease after discontinuation of prophylaxis. Although intravenous ganciclovir is effective standard treatment of tissue-invasive CMV disease in renal transplant recipients, emergence of ganciclovir-resistant CMV strain as a result of mutation of CMV genes involved in viral metabolism of CMV such as UL97 or UL54 is reported in renal transplant recipients.

Comparison and evaluation study on two hepatitis B virus DNA quantitative tests / 中华检验医学杂志

Objective To evaluate the two methods for quantitative hepatitis B virus (HBV) DNA test. Methods The Hybrid Capture II system from Digene Co. and Real Time PCR fluorimetry quantitative HBV DNA test kit from PIJI Bio Technical Development Company Ltd. were used to detect sera HBV DNA in sera of patients with chronic hepatitis B. Results With the two quantitative methods, the positive rates of HBV DNA were 98 6% and 94.6% respectively. The concordant rate of these two methods was 93.2%. In assessing the sensitivity of the two kits with a serial of 10 fold diluted patients′s sera, PG kit could test 10 -7 of diluted serum, and HC II could test 10 -6 . The test value of HC II was more accurate in quantitation than PG kit especially in higher titer of HBV DNA. Whereas in the case of low titer of HBV DNA, the deviation of test value increased in both two methods. For monitoring the anti virus effect with quantitative HBV DNA in CHB patients, these two methods had same sensitivity, and the test value of HBV DNA had the same trend of change. Conclusion HC II system and PG test kit were sensitive and reliable, and they were worthy of monitoring the anti virus efficacy and of clinical practice.