Four major envelope proteins of white spot syndrome virus bind to form a complex.

Early events in white spot syndrome virus (WSSV) morphogenesis, particularly the formation of viral membranes, are poorly understood. The major envelope proteins of WSSV are VP28, VP26, VP24, and VP19. Our previous results indicated that VP28 interacts with VP26 and VP24.

Lipid of white-spot syndrome virus originating from host-cell nuclei.

The hypothesis that white-spot syndrome virus (WSSV) generates its envelope in the nucleoplasm is based on electron microscopy observations; however, as yet there is no direct evidence for this. In the present study, the lipids of WSSV and the nuclei of its host, the crayfish Procambarus clarkii, were extracted and the neutral lipid and phospholipid contents were analysed by high-performance liquid chromatography, thin-layer chromatography and gas chromatography/mass spectrometry. Phosphatidylcholine (PC) and phosphatidylethanolamine comprised 62.

[Scanning electron microscopic observation of erythrocytes and endothelial cells of electrified death rabbits].

Objective: To identify electrified death of animals without electric marks.

Methods: Two animal models (model A and model B) of electrified death without electric marks were established. Model A: right anterior limb and left hind limb of rabbits were soddened by 0.

Application of spectrophotometry to evaluate the concentration of purified White Spot Syndrome Virus.

White Spot Syndrome Virus (WSSV) is a highly virulent pathogen of shrimp. In previous work, a simple and efficient method has been established in our laboratory to purify intact WSSV virions from infected crayfish tissues. To perform studies of WSSV infection mechanism, pathogenesis and gene function by using this purified virion, quantitative assay for the virus becomes increasingly important.

Identification and functional analysis of LsMNPV anti-apoptosis genes.

Three anti-apoptosis genes, Ls-iap2, iap3 and p49 were found in Leucania separata multiple nuclear polyhedrovirus. Amino acid sequence homology of Ls-IAP2 and Ls-IAP3 with Op-IAP2 and Op-IAP3 from Orgyia pseddotsugata MNPV were 20% and 42%, while that of Ls-P49 is 28% with Sl-P49 from Spodoptera littorolis MNPV. Ls-IAP2 contains one baculoviral IAP repeat (BIR) domain followed by a RING domain, while Ls-IAP3 contains two BIRs and a RING.

[Postmortem changes of liver phosphofructokinase-2 level in rats following different causes of death].

Objective: To study the changes of liver phosphofructokinase-2 (PFK-2) level at different postmortem intervals as well as due to different causes of death.

Methods: One hundred and five rats were randomly divided into 3 groups and the rats were sacrificed by either bleeding, suffocating, and neck breaking, respectively. The content of liver PFK-2 at 0, 1, 2, 4, 8, 12 and 24 hours following death were studied using immunohistochemishtry and image analysis.

Comparison between gene expression of conidia and germinating phase in Trichophyton rubrum.

Trichophyton rubrum is a dominating superficial dermatophyte, whose conidial germination is correlated to pathopoiesis and a highly important developmental process. To investigate the changes of physiology, biochemistry and cytology during the germination, we selected 3364 function identified ESTs from T. rubrum cDNA library to construct cDNA microarrays, and compared the gene expression levels of conidia and germinating phase.

Analysis of proteins that interact with nucleocapsid protein of SARS-CoV using 15-mer phage-displayed library.

Analysis of proteins that interact with N protein of SARS-CoV using 15-mer phage-displayed library will help to explore the virus pathogenesis and to develop new drugs and vaccines against SARS. In this study, we cloned, expressed and purified N protein of SARS-CoV. This 46-kD N protein was verified by SDS-PAGE and Western-blot.

[Improvement of baculovirus expression system and purification of IL-6 protein expressed in insect cells].

Based on site-specific transposition of an expression cassette into a baculovirus shuttle vector (Bacmid) which propagated in Escherichia coli, the Bac-to-Bac System provides a rapid and efficient method to generate recombinant baculoviruses and is widely used for high level expression of heterologous proteins. And the efficiency of recombinant baculovirus infecting cells plays an important role on the protein expression. In this study, we introduced an EGFP expression cassette driven by polyhedrin promoter into the p74 locus of Bacmid by homologous recombination.

[Expression of c-fos protein after muscle contusion in experimental rats].

Objective: To study the relationship between the expression of c-Fos protooncogene and skeleton muscle contusion of rats, and to search for a sensitive marker of timing for skeleton muscle contusion.

Methods: Fifty Sprague-Dawley rats were randomly distributed into control group and experimental groups. The expression of c-fos was microscopically observed by immunohistochemical method.

Co-involvement of the mitochondria and endoplasmic reticulum in cell death induced by the novel ER-targeted protein HAP.

HAP (a homologue of the ASY/Nogo-B protein), a novel human apoptosis-inducing protein, was found to be identical to RTN3. In an earlier study, we demonstrated that HAP localized exclusively to the endoplasmic reticulum (ER) and that its overexpression could induce cell apoptosis via a depletion of endoplasmic reticulum (ER) Ca(2+) stores. In this study, we show that overexpression of HAP causes the activation of caspase-12 and caspase-3.

[A study on expression of caspase-8 in organs of rats after electrical injury at antemortem or postmortem].

Objective: This study was conducted to detect the expression of caspase-8 in organs of rats after The electrical injury so as to elucidate whether caspase-8 is useful in identifying electrical lesion.

Methods: experiment included two parts. In the first part (the antemortem electrical injury part), thirty-five healthy male SD rats were randomly divided into seven groups (n = 5 per group), i.

[Expression of caspase-8 after brain injury from fluid percussion in rats].

Objective: To provide the evidence of the relationship between brain injury and the time of injury.

Methods: Rats were contused on brain by fluid percussion, then were killed after injury for 15 min, 30 min, 1,3,6,12 h, and 1,4,7,14 d respectively. The expression of caspase-8 were detected by immunohistochemical technology on rat brain section and the results were assessed by image analysis system in the cerebral cortex, thalamus, and hippocampus.

Tumor-targeted therapy with a conditionally replicating mutant of HSV-1 induces regression of xenografted human hepatomas.

The selectively oncolytic effects of mtHSV, a HSV icp34.5 mutant with lacz gene insertion, on several tumor cells in vitro and its antitumor effects by the intratumoral (IT) route to nude mice loaded the human hepatoma xenografts were explored. The mtHSV could conditionally replicate in and lyse Hep-3B (human hepatoma cells), Hep-2 (human larynx cancer cells) and SPC-A1 (human lung cancer cells), but not MRC-5 (human fibroblast cells).

[The expression of Fas after fluid percussion brain injury in rats].

Objective: To inquire into the pathologic diagnosis and the dating of injuries of light closed encephalon injury.

Methods: Wistar rats were hurt by fluid percussion, and were killed respectively at 15 min, 30 min, 1 h, 3 h, 6 h, 12 h, 1 d, 4 d, 7 d, and 14 d after injury. The expression of Fas in the cerebral cortex, thalamus, and hippocampi was detected by immunohistochemistry and the results were assessed by image analysis system.

Effective gene-virotherapy for complete eradication of tumor mediated by the combination of hTRAIL (TNFSF10) and plasminogen k5.

Virotherapy with oncolytic viruses is a highly promising approach for cancer therapy. To improve further the therapeutic effect of oncolytic viruses, therapeutic genes have been incorporated into these types of vectors. In this study, we have inserted hTRAIL (approved gene symbol TNFSF10) into the ZD55 vector, which was based on deletion of the adenoviral E1B 55-kDa gene and could replicate in and lyse p53-deficient tumors.

Identification of single-chain antibody fragments specific against SARS-associated coronavirus from phage-displayed antibody library.

To develop early diagnostic reagents, effective vaccines, and even drugs against SARS-associated coronavirus (SARS-CoV), the human single fold single-chain antibody fragments, (scFv) libraries I+J (Tomlinson I+J) were used to identify novel scFvs, which can specifically bind to SARS-CoV. Interestingly, two scFvs (B5 and B9) exhibited higher binding specificity to SARS-CoV with the OD(450) value 0.608 and 0.

Construction of a transposon-mediated baculovirus vector Hanpvid and a new cell line for expressing barnase.

In this study we developed the transposon-mediated shuttle vector 'Hanpvid', which composed of HaNPV (Heliothis armigera nuclear polyhedrosis virus) genomic DNA and a transposon cassette from Bacmid of Bac-to-Bac system. Hanpvid replicates in E. coli in the same way as Bacmid and retains infective function in cotton bollworm cells (Hz-AM1).

[A study on expression of hsp70 mRNA in the early stage of diffuse brain injury in rats].

Objective: To evaluate the changes of hsp70 mRNA in neuron and glia of rat suffering early diffuse brain injury.

Methods: SD rats were used in the replication of the animal pattern of diffuse brain injury. In situ hybridization technique and image analysis technique were applied in detecting the hsp70 mRNA in the rat's cerebral cortex, thalamus and brain stem.

Generation and characterization of a novel single-chain antibody fragment specific against human fibrin clots from phage display antibody library.

A novel single-chain fragment variable (scFv) antibody was developed directed against human fibrin clots by using the human single fold scFv libraries I+J (Tomlinson I+J). Three positively binding scFvs were evaluated by scFv-phage enzyme-linked immunosorbent assay (ELISA) and DNA sequencing. Then the positive scFv was expressed in soluble form in Escherichia coli HB2151 and purified by immobilized metal affinity chromatography (IMAC) with a yield of about 1 mg/l, the expression of soluble scFv was verified by Western blot analysis.

[Establishment of stable expression cell lines for HBsAg variants and analysis of antigenicity].

Objective: To study the mechanism of hepatitis B virus infected patients who is negative for HbsAg.

Methods: DNA sequences of 46 patients were analyzed. In these patients, HBsAg was negative but HBV DNA was positive and six new HBsAg variants were identified.

[Changes of connexin 43 in rabbit with early myocardial ischemia].

Objective: To reveal gap junction connexin 43 (Cx43) in rabbit with early myocardial ischemia.

Methods: The left coronary arteries were ligated to create early myocardial ischemia. Cx43 was visualized by use of SP immunohistochemical method (IHC) and Colloidal gold of the electron microscopy, and a comparison was made between the experiment group and the control group.

Identification of two novel fibrinolytic enzymes from Bacillus subtilis QK02.

Two fibrinolytic enzymes (QK-1 and QK-2) purified from the supernatant of Bacillus subtilis QK02 culture broth had molecular masses of 42,000 Da and 28,000 Da, respectively. The first 20 amino acids of the N-terminal sequence are AQSVPYGISQ IKAPALHSQG. The deduced protein sequence and its restriction enzyme map of the enzyme QK-2 are different from those of other proteases.

Bcl10 protein can act as a transcription activator in yeast.

The Bcl10 gene has recently been cloned from the chromosomal translocation t(1:14) (p22; q32) in a low-grade mucosa-associated lymphoid tissue (MALT) lymphoma, and was implicated in the pathogenesis of this and several other tumor types. In yeast two hybrid systems, when it was fused to Gal4 DNA-binding domain of pGBT9, this fusion protein can activate the expression of reporter genes without Gal4-AD domain. Through deletion assay and secondary structure prediction, we found that the N-terminal of Bcl10 contributed more to activation than the C-terminal.

Conditional replication of a recombinant adenovirus studied using neomycin as a selective marker.

An E1B-defective adenovirus, named r2/Ad carrying the neo expression cassette, was constructed by homologous recombination. The construction, selection (using neomycin as a selective marker), and propagation of the recombinant virus was performed in human embryonic kidney 293 cells (HEK 293). An in vitro study demonstrated that this recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as human glioma tumor cells (U251) and human bladder cells (EJ), but not in some cells with functional p53, such as human adenocarcinoma cells (A549) and human fibroblast cells (MRC-5).