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Biomedical subjects

W Z Jiang

Publications and source records attributed to W Z Jiang.

6 recordsLinked to original sources

Engineering and functional evaluation of a single-chain antibody against HIV-1 external glycoprotein gp120.

The HIV-1 envelope glycoprotein surface subunit gp120 is an attractive target for molecular intervention. This is because anti-HIV-1 gp120 neutralizing antibodies display the potential ability to inhibit HIV-1 infection. The present investigation describes the construction of a genetically engineered single chain antibody (scFv102) against HIV-1 gp120, its expression and functional evaluation. The parental hybridoma cell line (102) produces an immunoglobulin directed against the conserved CD4-binding region of gp120. cDNAs encoding the variable regions of the heavy (V(H)) and light (V(L)) chains were prepared by reverse transcription PCR and linked together with an oligonucleotide encoding a linker peptide (Gly(4)Ser)(3) to produce a single chain antibody gene. The resulting DNA construct was cloned into a prokaryotic expression vector (pET28) and recombinant scFv102 was expressed in Eserichia coli as an insoluble protein. The denatured scFv102 was refolded and purified by immobilized metal ion affinity chromatography. Purified scFv102 had the same specificity as the intact IgG in immuno-blotting assays and immuno-fluorescence (IF) detection, but ELISA analyses demonstrated the affinity of scFv102 to be 5-fold lower than that of the parental monoclonal antibody. In neutralization assays, scFv102 at concentrations lower than 40 microg/ml exhibited efficient interference with viral replication and inhibition of viral infection (90%) across a range of primary isolates of subtype B HIV-1. These results suggest that the constructed anti-HIV-1 gp120 scFv102 has good biological activity and can potentially be used for in vitro diagnostic and in vivo therapeutic applications.

Amino Acid Sequence↗

Cloning and expression of the external-glycoprotein gene mutant from HIV-2 in the methylotrophic yeast Pichia pastoris and identification of the glycoprotein.

To achieve high-level expression of HIV-2(ROD) external glycoprotein gp105 in Pichia pastoris, the gp105 gene mutant tP1, with the 5' non-functional region of the gp105 gene removed, was obtained by PCR amplification and was cloned into secreted expression vector pHILS1. The His(+)Mut(s) recombinant P. pastoris strain was screened by PCR and induced by methanol. SDS/PAGE and Western-blot analyses showed that mutation of the low-usage codon AGG into synonymous codon CGA and the introduction of the optimal codon TTC made P. pastoris overexpress tP1, an 85 kDa heterologous glycoprotein that was secreted into the medium and recognized specifically by HIV-2 polyclonal antibody. The recombinant strain GS115/tP1 had excellent genetic stability in terms of the properties of growth and expression of gp105, and seven out of 58 recombinant stains with a yield of 29% were selected to be used for further purification of gp105.

Blotting, Western↗

Lung cancer and indoor air pollution in Xuan Wei, China.

In Xuan Wei County, Yunnan Province, lung cancer mortality is among China's highest and, especially in females, is more closely associated with indoor burning of "smoky" coal, as opposed to wood or "smokeless" coal, than with tobacco smoking. Indoor air samples were collected during the burning of all three fuels. In contrast to wood and smokeless coal emissions, smoky coal emission has high concentrations of submicron particles containing mutagenic organics, especially in aromatic and polar fractions. These studies suggested an etiologic link between domestic smoky coal burning and lung cancer in Xuan Wei.

China↗

The epidemiology of lung cancer in Xuan Wei, China: current progress, issues, and research strategies.

In Xuan Wei, a rural Chinese county of about one million people, females' annual lung cancer mortality is China's highest, and males' is among China's highest. Xuan Wei's very high indoor air pollution levels (sometimes exceeding 20 mg/m3), residentially stable population, relatively uncomplicated lifestyle, and wide geographic variation in lung cancer mortality render it highly amenable to quantitative, interdisciplinary investigation of chemical carcinogens due to indoor air pollution. To date, epidemiologic findings reveal a closer association of lung cancer with the indoor burning of "smoky" coal (as opposed to "smokeless" coal or wood) than with tobacco use or occupation. Current aerometric, chemical, and toxicologic findings tend to confirm this association, though the specific carcinogenic constituents of Xuan Wei indoor air pollution have not yet been determined. Chinese and American investigators are conducting interdisciplinary field and laboratory investigations to quantify the lung cancer risk attendant on indoor air pollution relative to other factors, to measure and compare the characteristics of pollution from different Xuan Wei fuels, to determine the relative etiologic importance of pollution composition and concentration, and to develop quantitative relationships between air pollution dose and lung cancer risk.

Adult↗

Consulting with radiologists outside the hospital by using Java.

A Java-based teleradiology system that makes use of the Internet has been developed. Using this system, an on-call, off-duty radiologist can make diagnoses and perform consultations easily by reviewing the transferred images at home. The image accessibility of the system allows a hospital with such a system to assist an affiliated rural hospital without a full-time radiologist. The system consists of three components: the image server subsystem, the database server subsystem, and the client subsystem. All client subsystems of the system are designed to be symmetric. Users may retrieve images, manipulate them, and perform remote consultations. In addition, a user may add annotations to an image area of interest. Screen synchronization is achieved by means of the command-passing technique and local command execution to reduce the network bandwidth and transmission demands; user interaction is achieved by means of a synchronized indicator for pointing out areas of interest and dialog windows for conversation. Because Java programs can run on heterogeneous platforms, the need for system maintenance and user training is minimized. Although the drawback of Internet bandwidth still exists, the system enables platform-independent teleradiology via the Internet and allows easy and cost-effective remote consultation.

Humans↗