Preparation and application of monoclonal antibodies against Herpes simplex virus-1 / 中国免疫学杂志

Objective:To prepare and screen monoclonal antibodies against Herpes simplex virus-1(HSV-1),and develop a double antibody sandwich quantitative enzyme-linked immunosorbent assay( Q-ELISA) for detection of HSV-1 particle. This method was used to control the quality of viral particle in the developing and manufacturing process of HSV-1. Methods: BALB/c mice was immunized with HSV-1 to prepare monoclonal antibodies. A double antibody sandwich Q-ELISA was developed to determine concentration of HSV-1 particle,which was based on the neutralizing monoclonal antibody 1F6 as capture antibody,and 2B1 as HRP-conjugated antibody. The performance of the reagent was evaluated,including specificity,sensitivity,precision,accuracy and linear. And the relation between the amount of virus detected by this method and the virus titer was analyzed by regression analysis method. Results: The Q-ELISA for HSV-1 particle was developed. The quantitation scope was 0. 125-2 μg/ml, the coefficient correlation was 0. 995 5, the limit of detection was 0. 125 μg/ml, the recovery was between 85. 6% and 107. 1%, the variation coefficient was lower than 10%, and the reagent does not react with other samples except HSV-1 antigen. This method has a good correlation with virus titer. Conclusion:The Q-ELISA for HSV-1 particle was successfully developed,which provide a new approach for rapid and quantitative detection of HSV-1 antigen.

Detection of mast cell degranulation by fluorescent molecular diffusion dynamic measurement / 中国病理生理杂志

AIM:To establish a rapid in vitro method for mast cell degranulation tracing by raster image corre-lation spectroscopy (RICS).METHODS:RBL-2H3, a basophilic granulocyte mast cell line transfected with CD 63-GFP plasmid, was used for evaluating the methods , including β-hexosaminidase ( HEX) colorimetric assay, scanning electron microscopy (SEM) and RICS in the detection of mast cell degranulation .The sensibilities of these methods were com-pared.RESULTS:The sensitivities of β-HEX colorimetric assay and SEM were 5 mg/L and 3.9 ×10 -2 mg/L, respec-tively.RICS detection showed obvious decrease in the diffusion coefficient at dose of 3.9 ×10 -2 mg/L.CONCLUSION:Fluorescent molecular diffusion dynamic measurement can be used for rapid tracing of allergic substances in vitro.Accord-ing to the results, RICS can achieve nearly the same extent of sensitivity as the SEM does and is far more sensitive than β-HEX colorimetric assay.Compared with SEM, RICS has several advantages: it is faster, simpler and cheaper; it can be used in living cells;it is more suitable for rapid in vitro allergenic compounds tracing .Therefore, RICS is applicable in clinic allergic antigen screening and may also be used in pharmaceutical quality control .

A novel pattern recognition protein of the Chinese oak silkmoth, Antheraea pernyi, is involved in the pro-PO activating system.

In this paper, we firstly reported a C-type lectin cDNA clone of 1029 bps from the larvae of A. Pernyi (Ap-CTL) using PCR and RACE techniques. The full-length cDNA contains an open reading frame encoding 308 amino acid residues which has two different carbohydrate-recognition domains (CRDs) arranged in tandem. To investigate the biological activities in the innate immunity, recombinant Ap-CTL was expressed in E. coli with a 6-histidine at the amino-terminus (Ap-rCTL). Besides acted as a broad-spectrum recognition protein binding to a wide range of PAMPs and microorganisms, Ap-rCTL also had the ability to recognize and trigger the agglutination of bacteria and fungi. In the proPO activation assay, Ap-rCTL specifically restored the PO activity of hemolymph blocked by anti- Ap-rCTL antibody in the presence of different PAMPs or microorganisms. In summary, Ap-rCTL plays an important role in insect innate immunity as an pattern recognition protein.

Purification and characterization of a 1,3-ß-D-glucan recognition protein from Antheraea pernyi larve that is regulated after a specific immune challenge.

Pattern recognition receptors are known to participate in the activation of Prophenoloxidase system. In this study, a 1,3-ß-D-glucan recognition protein was detected for the first time in Antheraea pernyi larvae (Ap-ßGRP). Ap-ßGRP was purified to 99.9% homogeneity from the hemolymph using traditional chromatographic methods. Ap-ßGRP specifically bind 1,3-ß-D-glucan and yeast, but not E. coli or M. luteus. The 1,3-ß-D-glucan dependent phenoloxidase (PO) activity of the hemolymph inhibited by anti-Ap-ßGRP antibody could be recovered by addition of purified Ap-ßGRP. These results demonstrate that Ap-ßGRP acts as a biosensor of 1,3-ß-Dglucan to trigger the Prophenoloxidase system. A trace mount of 1,3-ß-D-glucan or Ap-ßGRP alone was unable to trigger the proPO system, but they both did. Ap-ßGRP was specifically degraded following the activation of proPO with 1,3-ß-Dglucan. These results indicate the variation in the amount of Ap-ßGRP after specific immune challenge in A. pernyi hemolymph is an important regulation mechanism to immune response.

Analysis of citation rate of scientific papers published in Medical Journal of Chinese People's Liberation Army / 解放军医学杂志

Objective To evaluate the academic level and the popularity of Medical Journal of Chinese People's Liberation Army. Method According to the information of Chinese Medical Citation Index(CMCI), The amount and distribution of the original in Medical Journal of Chinese People's Liberation Army cited by the journal included by CMCI were analyzed. Result The citation rate of published articles was 36.54%, and the average of original articles cited by other researchers was 2.70. The distribution of the most frequently cited authors covered 24 provinces, with Beijing, Shanghai and Guangdong Province in the lead in research work relevant to military medicine. The published papers were cited by 451 journals, and self-citing rate was 0.20. Conclusion Original medical scientific articles of high quality have been published in the Medical Journal of Chinese People′s Liberation Army. It have become one of the most important information resource for the medical researchers in the army and one of the important medical journal in the country.

Comparison of Ability of BACTEC 9120 Blood Culture System to Detect Fungemia with Four Kinds of Blood Culture Bottles / 中华医院感染学杂志

OBJECTIVE To discuss the best scheme for fungemia detection by analyzing and comparing the ability and performance characteristics of the BACTEC 9120 automated blood culture system and 4 kinds of blood culture bottles in the detection of simulated fungemia.METHODS Simulated blood culture was produced using 65 fungi isolates from clinical specimens and BACTEC Plus Aerobic/F,Plus Anaerobic/F,Peds Plus/F and Myco/F Lytic blood culture bottles and detected by BACTEC 9120 automated blood culture system.The final inoculum densities were 1-5 CFU/ml blood.The(time to detection TTD) of simulated blood culture with different concentrations of suspension produced using 2 kinds of standard strains and 4 kinds of blood culture bottles was compared.RESULTS From the 260 bottles in this study 216 had growth detected by the BACTEC 9120 blood culture system.The positive rates of BACTEC Plus Aerobic/F and Anaerobic/F,which were 90.77%and 41.54%,respectively,were significantly(P