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Biomedical subjects

Bao Zhang

Publications and source records attributed to Bao Zhang.

At least 37 records · Page 2Linked to original sources
PubMed2004

[Relationship of serum levels of HGF and MMP-9 with disease activity of patients with systemic lupus erythematosus].

OBJECTIVE: To determine the relationship of serum hepatocyte growth factor (HGF) and matrix metalloproteinase-9 (MMP-9) levels with the disease activity of systemic lupus erythematosus (SLE). METHODS: Serum levels of HGF and MMP-9 were measured by ELISA in 36 patients with SLE and 30 healthy subjects as controls. RESULT: (1)Significantly increased serum level of HGF was found in SLE patients as compared with that in healthy controls (P<0.001), but serum level of MMP-9 in SLE patients decreased (P<0.001). Serum level of HGF was significantly decreased after treatment in SLE patients (P<0.05), but serum level of MMP-9 was increased (P<0.05). (2)Serum level of HGF was markedly higher in patients with active disease (24 cases) than those with inactive disease (P<0.05), but serum level of MMP-9 was lower (P<0.05). (3)Significantly increased serum level of HGF was found in patients with renal damage (16 cases) than those without (P<0.001), but serum level of MMP-9 was lower in patients with renal damage than those without (P<0.005). (4)Serum level of HGF was higher in patients with arthritis (23 cases) than those without (P<0.01), but serum level of MMP-9 had no significant difference in two groups (P>0.05). (5)The area of ROC curve was 0.707 and the sensitivity was 66.7% if serum level of HGF was served as diagnostic standard. The area of ROC curve was 0.984 and the sensitivity was 97.2% if serum level of MMP-9 was served as diagnostic standard. The sensitivity was 66.7% (24/36) if the two markers were examined simultaneously. CONCLUSION: The data suggest that HGF and MMP-9 could be involved in the pathogenesis of SLE, and serum levels of HGF and MMP-9 be used as markers to monitor disease activity, renal damage, disease progression and amelioration in SLE.

Adolescent↗
PubMed2004

[Amplification and cloning of the N gene of SARS-associated coronavirus].

OBJECTIVE: To amplify and clone the N gene of severe acute respiratory syndrome-associated coronavirus. METHOD: Using primer Premier 5.0 software, two pairs of nested PCR primers were designed to amplify the N gene. After purification, the amplified products were cloned into pMD18-T vectors, and the positive clones with the inserted fragments were identified by sequence analysis. RESULTS: The amplified products was about 1 375 bp in length, and sequence analysis demonstrated that the N gene fragments had been successfully inserted into pMD18-T vectors. CONCLUSION: The successful amplification and cloning of N gene facilitates further investigation of the expression of the N protein and study of its structure and functions.

Base Sequence↗
PubMed2004

[Preparation of the microarray for detecting hepatitis D virus].

OBJECTIVE: To prepare the microarray for detection of hepatitis D virus (HDV). METHOD: Several pairs of specific PCR primers were designed according to the conserved region of HDV genome. The DNA microarray were prepared by blotting the PCR products onto the surface of glass slides with the use of robotics, and restriction display PCR (RD-PCR) was employed to label the samples. RESULT: Sequences analysis showed that the products of PCR amplification were the specific gene fragments of HDV. Hybridization signals on the gene chip demonstrated good specificity and sensitivity of the microarray for HDV detection. CONCLUSION: Microarray-based clinical HDV detection can be sensitive and effective.

Base Sequence↗
PubMed2004

[DNA microarray for the detection of Yersinia pesits].

OBJECTIVE: To develop a DNA microarray technique for fast diagnosis of plague. METHODS: Restriction display polymerase chain reaction (RD-PCR) and hybridization with fluorescently labeled cy5 were employed for detecting the sample DNA of Yersinia pestis, Yersinia pseudotuberculosis and Yersinia enterocolitica. RESULTS: The prepared microarray is capable of distinguishing Yersinia pestis from Yersinia pseudotuberculosis and Yersinia enterocolitica in the same genus. CONCLUSION: The constructed DNA microarray is an effective diagnostic tool for the detection of Yersinia pestis.

Fluorescence↗
PubMed2004

[Co-expression of two spliceosomes of cyclin-dependent kinase-2 in SH-SY5Y cells].

OBJECTIVE: To investigate the expression of cyclin-dependent kinase-2 (CDK-2) gene in SH-SY5Y cells. METHODS: The expression of CDK-2 gene was examined with reverse transcriptional (RT)-PCR, and the PCR products underwent electrophoresis on non-denaturing poly-acrylamide gel (PAG) followed by silver staining. The separated and purified DNAs were ligated into pMD18-T vector, and the positive clones identified by sequence analysis. RESULTS: Two DNA bands were displayed on PAG, and the one with smaller molecular weight was less intensively stained. The sequences of the two clones indicated that both were products of CDK-2 gene. CONCLUSION: Two kinds of CDK-2 gene products are co-expressed in the SH-SY5Y cells, one of which lacks the fifth exon and is expressed at a low level.

CDC2-CDC28 Kinases↗
PubMed2004

Detection of hepatitis D virus by cDNA microarray method.

BACKGROUND: Viral hepatitis is considered a major public health problem in most areas of the world. In acute and chronic infections, hepatitis D virus (HDV) infection often leads to a more severe disease. This study was designed to prepare microarrays for HDV detection. METHODS: The specific primers of PCR were designed according to the conserved region of HDV. The cDNA microarrays were prepared by spotting PCR products onto the surface of glass slides by robotics. Restriction display PCR (RD-PCR) was used to label the samples. RESULTS: Sequences were aligned, and the results showed that the products of PCR amplification were the specific gene fragments of HDV. Hybridizing signals on gene chip showed the specificity and sensitivity in detecting HDV were satisfactory. CONCLUSION: Using PCR amplified products to construct gene chips for clinical diagnosis of HDV is a quick, simple and effective method.

Base Sequence↗
PubMed2004

[Effect of astrocyte-conditioned medium on tert-butyl hydroperoxide-induced apoptosis of PC12 cells].

OBJECTIVE: To investigate the effect of astrocyte-conditioned medium (ACM) on PC12 cell apoptosis induced by reactive oxygen species tert-butyl hydroperoxide (tbOOH). METHODS: PC12 cells attacked by tbOOH were cultured with Sprague Dawley rat cerebral cortex ACM, and the cell apoptosis was observed by fluorescent microscopy and transmission electron microscopy. The cell apoptosis rate was evaluated by flow cytometry and malondialdehyde (MDA) concentration measured using thiobarbituric acid (TBA)-reacting substances (TRABS). RESULT: TbOOH induced PC12 cell apoptosis, and ACM significantly decreased the apoptosis rate and MDA concentration in tbOOH-treated cells. CONCLUSION: ACM may increase the anti-oxidation capacity of PC12 cells and inhibit cell apoptosis induced by tbOOH.

Animals↗
PubMed2004

[Oligonucleotide microarray preparation using enhanced poly-L-lysine glass slides].

OBJECTIVE: To modify conventional poly-L-lysine coating for oligonucleotide microarray preparation so as to enhance the sensitivity of the microarray. METHOD: The proposed chemical approaches included silanizing the slides with 3-glycid-oxypropyltrimethoxysilane (GOPS) after cleaning, followed by slide coating with polymers (poly-L-lysine) that was covalently bound to the modified glass. Subsequent attachment of the oligonucleotide to the modified slide surface was achieved after 1,4-phenylene diisothiocyanate (PDITC) activation of the surface. Various experiments were carried out, such as the immobilization efficiency and hybridization assays to test the modified slides, which were then used tentatively in the preparation of microarrays for SARS coronavirus detection. RESULTS: The improved surface had high immobilization efficiency, good uniformity and satisfactory hybridization efficiency, better than those slides with conventional poly-L-lysine coating. In addition, such modified slides rendered the microarrays more resistant to consecutive probing/stripping cycles. CONCLUSION: The modified slide surface is satisfactory to immobilize unmodified oligonucleotide by covalent binding, which enhances not only the sensitivity of the prepared oligonucleotide microarray but also the binding of the oligonucleotide to the slide surface.

DNA, Viral↗
PubMed2003

Possibility of using DNA chip technology for diagnosis of human papillomavirus.

To explore the application of DNA chip technology for the detection and typing of Human Papillomavirus (HPV), the HPV6, 11, 16 and 18 gene fragments were isolated and printed onto aminosilane-coated glass slides by a PixSys 5500 microarrayer as probes to prepare the HPV gene chips. HPV samples, after being labeled with fluorescent dye by restriction display PCR (RD-PCR) technology, were hybridized with the microarray, which was followed by scanning and analysis. The experimental condition for preparing the HPV gene chips was investigated, and the possibility of HPV genotyping using gene chips was discussed. The technique that was established in this study for preparing HPV gene chips is practical. The results of the present study demonstrated the versatility and inspiring prospect of using this technology to detect and genotype HPV.

DNA Probes↗
PubMed2003

Gene expression study of Saccharomyces cerevisiae with the Agilent 2100 bioanalyser.

This study explores the restriction display-polymerase chain reaction (RD-PCR) application of a new chip-based nucleic acid analysis system (Agilent 2100 bioanalyser) in a gene differential expression study. Total RNAs is extracted from Saccharomyces cerevisiae, double-stranded complementary DNA (cDNA) is synthesised by reverse transcription from the purified messenger RNA (mRNA), RD-PCR conducted to obtain the cDNA fragments and bioanalyser and agarose gel electrophoresis compared for the analysis of RD-PCR products. The bioanalyser proved to be faster and more sensitive in separating and detecting gene fragments, and was also able to compare different gene fragments quantitatively. Using this technology, comparison of several differential gene fragments is performed.

DNA↗
PubMed2003

Cloning and sequence analysis of Bacillus thuringiensis gene fragments isolated by restriction digest PCR.

OBJECTIVE: To clone and analyze Bacillus thuringiensis gene fragments isolated by restriction digest PCR (RD-PCR). METHOD: Specific primers were designed to amplify the genes of Bacillus thuringiensis israelensis (Bti), and the PCR products were classified and re-amplified by RD-PCR to obtain the fragments for subsequent purification and cloning into the pMD18-T vectors, followed by rapid identification. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. RESULTS: Sequence analysis showed that all the fragments amplified were Bti genes. CONCLUSION: RD-PCR is reliable in breaking down large gene fragments into confined and shorter gene fragments for preparing microarray probes.

Bacillus thuringiensis↗
PubMed2003

[Gene sequence analysis of SARS-associated coronavirus by nested RT-PCR].

OBJECTIVE: To explore an effective means for the detection of Severe Acute Respiratory Syndrome (SARS)- associated coronavirus. METHODS: The RNAs of the virus contained in the sputum samples from established SARS patients were extracted and reversely transcripted, followed by nested PCR using the reversely transcripted cDNA as the template. The PCR products were cloned then into the pMD18-T vectors, followed by sequence analysis. RESULTS: Specific fragments were amplified from the sputum samples of SARS patients, which were confirmed by DNA cloning and sequencing to belong to SARS-associated coronavirus. The Result of Blast shows only the difference in one nucleic acid from the TOR2 strain of SARS-associated coronavirus. CONCLUSION: Sequence analysis has confirmed the existence of SARS-associated coronavirus in the sputum samples of SARS patients, and nested RT-PCR is a quick, easy, and convenient way for the detection of the virus.

Base Sequence↗
PubMed2003

Rapid preparation of DNA microarray using PCR for hepatitis B and D virus detection.

OBJECTIVE: To prepare DNA microarray for detecting both hepatitis B and D virus as (HBV and HDV). METHODS: With the assistance of Oligo6.4 software, specific PCR primers targeting the conserved region of HBV and HDV were designed. The PCR products were purified and cloned into the pMD18-T vectors, followed by rapid identification. The recombinant plasmids were then extracted from positive clones and the target gene fragments underwent sequence analysis. RESULTS: The gene fragments of HBV and HDV were obtained by PCR, which were confirmed by sequence analysis to be specific gene fragments of HBV and HDV. CONCLUSION: Using PCR amplification products to prepare the DNA microarray is quick, simple and effective.

Hepatitis B virus↗
PubMed2003

[Tumor infiltrating dendritic cells and Mucin1 gene expression in benign prostatic hyperplasia and prostate cancer].

OBJECTIVE: To study the expression of Mucin1 gene and tumor infiltrating dendritic cells(TIDC) in the tissues of benign prostatic hyperplasia (BPH) and prostate cancer. METHODS: Mucin1 and TIDC were detected in 20 specimens of BPH and 30 specimens of prostate cancer by immunohistochemistry SP method. RESULTS: MUC1 expressed in both prostate cancer and BPH. The staining patterns were significantly associated with tumor pathological grade (P < 0.001). The number of TIDC was negatively correlated with tumor pathological grade, the higher the grade, the smaller the number of TIDC (P < 0.001). CONCLUSIONS: The expression pattern of MUC1 and the number of TIDC could be considered as useful markers to evaluate the malignant degree and prognosis of prostate cancer. The decrease of TIDC plays an important role in tumor immune evasion and immune tolerance. Highly expressed MUC1 could lead to the failure of hormonal treatment for prostate cancer, and contribute much to tumor infiltration and metastasis.

Antigens, Neoplasm↗
PubMed2003

[Study on development of DNA microarrays for human papillomavirus (HPV) diagnosis].

To study the technology for establishing DNA microarrays for the diagnosis of HPV. HPV6, 11, 16 and 18 gene fragments were isolated and printed onto aminosilane-coated glass slides by PixSys 5500 microarray printer as probes to prepare the HPV. HPV samples, after labeled with Cy3 or Cy5, were hybridaized with the microarray followed by scanning for analysis. The experimental condition for preparing the HPV gene chips was investigated and the possibility of HPV genotying using DNA microarrays was discussed. The technique established in this study for preparing HPV DNA microarrays is applicable and has potential clinical application significance.

Alphapapillomavirus↗
PubMed2002

[Purification of PCR products with cetyltrimethylammonium bromide].

OBJECTIVE: To establish a method for purifying PCR products with cetyltrimethylammonium bromide (CTAB). METHOD: Selective precipitation of the PCR product was performed using CTAB, which forms compound with DNA fragment in salt solution of appropriate concentration, but not with single strain oligonucleotide or dNTPs. The precipitation could be dissolved in 1.2 mol/L NaCl while the addition of ethanol caused the desired PCR product to precipitate so as to be recovered. RESULT: The primers and small-molecule dNTP could be effectively eliminated after this procedure. Although the output of the purification process reached only 80% that by current reagent kit, it reduces the cost to as low as 1/8 of that normally required by the kit. CONCLUSION: CTAB is applicable for purification of the PCR product.

Cetrimonium↗
PubMed2002

[Cloning of an expressed sequence tag with restriction display polymerase chain reaction].

OBJECTIVE: To isolate gene fragments from SH-SY5Y cells by way of restriction display polymerase chain reaction (RD-PCR). METHODS: Total mRNA was extracted from SH-SY5Y cells followed by synthesis of the single-strand cDNA with Oligo (dT18) as the anchored primer, and the second strand was synthesized by nick translation. The double strands were cleft with restriction enzyme Sau3A I and the fragments ligated with a universal adapter to be amplified with the universal primers and selected primers. The products were then ligated into the pMD18-T vector and sequenced. RESULTS: One of the sequenced clones was retrieved in the National Center for Biotechnology Information (NCBI) databases with Blast program. The results showed that the sequence possessed great similarity to one fragment of the 17th chromosome in the genome. Sequence analysis with GenScan software indicated that the EST might be one section of an unknown gene. CONCLUSION: RD-PCR provides simple and efficient approach for isolating EST from cells, and cDNA clone sequencing combined with bioinformatics analysis may be helpful in identifying new genes.

Base Sequence↗