Search PubMed⌕ Search

Biomedical subjects

Z Deng

Publications and source records attributed to Z Deng.

At least 109 records · Page 6Linked to original sources
PubMed1996

Partial purification of smooth muscle cell derived growth factor.

The serum free medium conditioned by cultured rabbit aortic smooth muscle cells was partially purified using ultrafiltration and heparin affinity chromatography. Incorporation of [3H]-thymidine (3H-TdR) into cell DNA was used to measure the mitogenic activity of the fractions from chromatography for NIH 3T3 fibroblasts. The molecular weight and the iso-electric point of these fractions were determined by NaDodSO4-polyacrylamide gel electrophoresis (SDS-PAGE) and iso-electric focusing, respectively. The results showed that the protein eluted in 1.0-1.6 mol/L NaCl from the heparin-Sepharose was mitogenic for 3T3 cells, and this protein had a molecular weight of 22.8-26.7 ku and an iso-electric point of about 4.6. The fact that the above-mentioned biochemical properties differed from that of PDGF, IGF and FGF suggests that this mitogenic protein may be a separate growth factor.

3T3 Cells↗
PubMed1996

Very low density lipoprotein and oxidized very low density lipoprotein induce monocyte chemotactic protein 1 in rabbit aortic smooth muscle cells.

OBJECTIVE: To understand whether very low density lipoprotein (VLDL) and oxidatively modified very low density lipoprotein (OX-VLDL) can stimulate vascular smooth muscle cells (SMCs) to express high level of monocyte chemotactic protein (MCP-1) mRNA. METHODS: LDL and VLDL were isolated from normal blood donors by density gradient ultracentrifugation and oxidatively modified by adding CuCl2 and incubation at 37 degrees C for 24 hours. Rabbit aortic SMCs were cultured by a substrate-attached explant method, and their total RNA was extracted by guanidinium isothiocyanate method. The expression of MCP-1 mRNA in SMCs was examined by dot blotting analysis using a probe of gamma-22P-end-labeled 35-mer oligonucleotide. RESULTS: Dot blotting analysis showed that cultured SMCs can express MCP-1 mRNA and the level of MCP-1 mRNA was increased significantly in SMCs after 24-hour coculture with VLDL (6-fold) and OX-VLDL (20-fold). CONCLUSIONS: VLDL and OX-VLDL both can induce MCP-1 mRNA expression in cultured rabbit aortic SMCs and the latter is more effective than the former.

Animals↗
PubMed1996

[Effects of oxidized low density lipoprotein and very low density lipoprotein on the expression of MCP-1 by monocytes].

Monocyte chemoattractant protein-1 (MCP-1) is a potent chemotatic factor for monocytes. This study aimed at examination of the effects of oxidized low density lipoprotein (OX-LDL) and oxidized very low density lipoprotein (OX-VLDL) on the expression of MCP-1 mRNA and protein by monocytes. The guanidinium isothiocyanate method was used to extract total RNA from monocytes which were exposed to LDL, OX-LDL, VLDL and OX-VLDL for 24 hours respectively and the monocyte derived conditioned media collected. MCP-1 mRNA expression in monocytes was examined by slot blot and Northern blot analysis using a gamma 32P-end-labelled 35 mer oligonucleotide probe of MCP-1. Meanwhile, MCP-1 protein in the conditioned media was examined by use of sandwich ELISA. It was found that monocytes from human peripheral blood can express MCP-1 mRNA and protein after 24 hours incubation at 37 degrees C and the expression of MCP-1 mRNA and protein was markedly increased by a 24 hour exposure to OX-LDL and OX-VLDL, whereas the expression of MCP-1 mRNA and protein was only sightly increased when exposed to LDL and VLDL. It is concluded that monocytes can express MCP-1 that induce additional monocyte migration into the subendothelial space, and that OX-LDL and OX-VLDL enhance this function by inducing stronger MCP-1 expression in monocytes.

Cells, Cultured↗
PubMed1995

Detection of Borrelia burgdorferi-specific antigen in antibody-negative cerebrospinal fluid in neurologic Lyme disease.

OBJECTIVE: To determine the potential of detection in CSF of specific Borrelia burgdorferi antigen, OspA, as a marker of infection in neurologic Lyme disease and compare this with the detection of antibody. DESIGN: CSF from 83 neurologic patients in an area highly endemic for Lyme disease was examined prospectively for (1) OspA by antigen capture ELISA and Western blot employing monoclonal antibodies, and for (2) B burgdorferi antibodies by ELISA. RESULTS: Of the 35 of 83 (42%) patients who were positive for OspA antigen in their CSF, 15 (43%) were antigen positive despite being antibody-negative in CSF. Seven of these 15 (47%) had otherwise normal routine CSF analyses. Six of these 15 (40%) patients met strict CDC surveillance criteria for Lyme disease; four (27%) patients had seroconversion coincident with new neurologic problems; and three (20%) with characteristic syndromes for Lyme disease were seronegative, but had complexed antibody to B burgdorferi. The final two patients (13%) were seropositive and had unexplained neurologic problems not characteristic of Lyme disease. CONCLUSIONS: B burgdorferi antigen can be detected in CSF that is otherwise normal by conventional methodology, and can be present without positive CSF antibody. Since CSF antigen implies intrathecal seeding of the infection, the diagnosis of neurologic infection by B burgdorferi should not be excluded solely on the basis of normal routine CSF or negative CSF antibody analyses.

Adult↗
PubMed1995

[Change of the guaning nucleotide-regulating protein in substantia gelatinosa of spinal cord of rats after unilateral transection of dorsal roots and electroacupuncture: an immunohistochemical observation].

The Subunit alpha o of guanine nucleotide-binding protein (rabbit polyclonal antiserum 9120, K,J. Chang. USA), in the areas of rat spinal cord was localized by immunohistochemical PAP methods, and the changes of alpha o-immunoreactivity (alpha o-IR) in the substantia gelatinosa (SG) were observed after unilateral transection of dorsal roots and electroacupuncture. Intense alpha o-IR was presented in Rexed lamina I to III of the dorsal horn, and the highest alpha o-IR in lamina (SG), which was found than that in the normal and control animals. Iateral spinal nucleus of rat revealed higher density of alpha o-IR containing fiber networks. Following unilateral transection of dorsal roots and pain threshold increased significantly after electroacupuncture applied at the point "Huantiao", alpha o-IR was markedly decreased than that in the normal and control group in those areas. The optic density (OD) of alpha o-IR were measured, which decreased more markedly in opration and electroacupuncture group than that normal and control group. The results mentioned above indicate that Go-protein in terminals of primary nociceptive neurons of SG decreases markedly after dorsal roots transected and electroacupuncture. It was inferenced that partial Go-protein probably originates from the primary sensory neurons, and the release of "inhibitory" of Go may increase (with weaker alpha o-IR). In a work, Go might play important roles on the primary afferents informational regulation and electroacupuncture analgesia of the regulation.

Animals↗
PubMed1994

Mapping of two phenol sulphotransferase genes, STP and STM, to 16p: candidate genes for Batten disease.

The cytosolic phenol sulphotransferase gene (STP) was mapped to a region of chromosome 16, within the interval defined by human-rodent somatic cell hybrid breakpoints CY160(D) and CY12, which contains FRA16E. YAC and cosmid clones from this 16p interval were screened for the presence of STP. Two non-overlapping cosmid contigs were identified which contain STP-like sequences. Sequencing of these STP-like sequences confirmed that STP is contained within contig 343.1 and maps proximal to FRA16E, and that a related sulphotransferase STM, encoding the catecholamine-sulphating enzyme, is contained within contig 55.4 and maps to the adjacent hybrid interval CY12-CY180A. Thus two phenol sulphotransferase genes (STP and STM) have been finely localised to chromosome 16p12.1-p11.2, to the same region as CLN3, the gene for Batten disease. Both genes are therefore candidate genes for Batten disease.

Animals↗
PubMed1994

Molecular cloning of the human nucleotide-excision-repair gene ERCC4.

ERCC4 was previously identified in somatic cell hybrids as a human gene that corrects the nucleotide-excision-repair deficiency in mutant hamster cells. The cloning strategy for ERCC4 involved transfection of the repair-deficient hamster cell line UV41 with a human sCos-1 cosmid library derived from chromosome 16. Enhanced UV resistance was seen with one cosmid-library transformant and two secondary transformants of UV41. Cosmid clones carrying a functional ERCC4 gene were isolated from a library of a secondary transformant by selecting in Escherichia coli for expression of a linked neomycin-resistance gene that was present in the sCos-1 vector. The cosmids mapped to 16p13.13-p13.2, the location assigned to ERCC4 by using somatic cell hybrids. Upon transfection into UV41, six cosmid clones gave partial correction ranging from 30% to 64%, although all appeared to contain the complete gene. The capacity for in vitro excision of thymine dimers from a plasmid by transformant cell extracts correlated qualitatively with enhanced UV resistance.

Animals↗
PubMed1994

Streptomyces lividans 66 contains a gene for phage resistance which is similar to the phage lambda ea59 endonuclease gene.

The DNA of wild-type Streptomyces lividans 66 is degraded during electrophoresis in buffers containing traces of ferrous iron. S. lividans ZX1, a mutant selected for resistance to DNA degradation, simultaneously became sensitive to phi HAU3, a wide-host-range temperate bacteriophage. A DNA fragment conferring phi HAU3 resistance was cloned; it contains a phage resistance gene whose deduced amino acid sequence is similar to the phage lambda Ea59 endonuclease. The S. lividans phi HAU3 resistance does not seem to be a classical restriction-modification system, because no host-modified phages able to propagate on the wild-type strain could be isolated. The cloned fragment did not make the host DNA prone to degradation during electrophoresis, indicating that the two phenotypes are controlled by different genes which were deleted together from the chromosome of ZX1.

Amino Acid Sequence↗
PubMed1994

Repeated polyketide synthase modules involved in the biosynthesis of a heptaene macrolide by Streptomyces sp. FR-008.

Genes for biosynthesis of a Streptomyces sp. FR-008 heptaene macrolide antibiotic with antifungal and mosquito larvicidal activity were cloned in Escherichia coli using heterologous DNA probes. The cloned genes were implicated in heptaene biosynthesis by gene replacement. The FR-008 antibiotic contains a 38-membered, polyketide-derived macrolide ring. Southern hybridization using probes encoding domains of the type I modular erythromycin polyketide synthase (PKS) showed that the Streptomyces sp. FR-008 PKS gene cluster contains repeated sequences spanning c. 105kb of contiguous DNA; assuming c. 5 kb for each PKS module, this is in striking agreement with the expectation for the 21-step condensation process required for synthesis of the FR-008 carbon chain. The methods developed for transformation and gene replacement in Streptomyces sp. FR-008 make it possible to genetically manipulate polyene macrolide production, and may later lead to the biosynthesis of novel polyene macrolides.

Anti-Bacterial Agents↗
PubMed1994

Development of a gene cloning system for Streptomyces hygroscopicus subsp. yingchengensis, a producer of three useful antifungal compounds, by elimination of three barriers to DNA transfer.

Streptomyces hygroscopicus 10-22 could not be transformed with any of the commonly used Streptomyces plasmid vectors and was resistant to plaque formation by the Streptomyces phages phi C31 and R4. Repeated selection resulted in the isolation of derivatives of S. hygroscopicus 10-22 that could be transformed with pIJ101- and pJV1-derived cloning vectors and of restriction-deficient derivatives that could accept DNA propagated in Streptomyces lividans 66. These new strains, which include three that still produce the original antibiotics, can be used as hosts for gene cloning. Insertion of nonreplicating vectors by homologous recombination and transposition of Tn4560 were demonstrated in S. hygroscopicus 10-22.

Antifungal Agents↗
PubMed1993

Smooth muscle myosin heavy chain locus (MYH11) maps to 16p13.13-p13.12 and establishes a new region of conserved synteny between human 16p and mouse 16.

The human smooth muscle myosin heavy chain locus (MYH11) was mapped by fluorescence in situ hybridization to the middle of the p arm of chromosome 16 using a genomic cosmid clone containing coding sequences of the gene as probe. Probe from coding sequence, when applied to Southern blots of a panel of hybrids containing different portions of human chromosome 16, localized the gene to 16p13.13-13.12. Coding sequence PCR primers, when used on the DNA from a CHO-mouse hybrid clone mapping panel informative for mouse chromosomes, showed that the gene was located on mouse chromosome 16. These results correct a recent assignment of MYH11 from 16q12.2 to the region of the 16p-arm inversion breakpoint seen in acute myelomonocytic leukemia (AMML) M4Eo and demonstrate that the conflicting data do not result from the presence of additional MYH genes on the q arm of the chromosome. Also, a new region of conserved synteny between human 16p and mouse 16 is established.

Animals↗
PubMed1993

Mapping of the phenol sulfotransferase gene (STP) to human chromosome 16p12.1-p11.2 and to mouse chromosome 7.

We have recently cloned a cDNA encoding the human phenol-preferring phenol sulfotransferase (P-PST) enzyme. An oligonucleotide primer pair based on the human STP (representing sulfotransferase, phenol-preferring) cDNA sequence was synthesized and was employed in polymerase chain reaction (PCR) amplification of human genomic DNA to identify a 525-bp DNA fragment. The DNA sequence of this portion of the STP gene, near the 5' end of the coding region, was determined. The amplified genomic fragment contained two small introns of 104 and 89 bp. When DNA samples from a human-hamster somatic cell hybrid panel were screened by PCR using these primers, only those hybrids that contained human chromosome 16 were positive for the 525-bp genomic fragment. To identify the specific region on chromosome 16 that contained the STP gene, PCR amplification reactions were performed on a human-mouse somatic cell hybrid panel containing defined portions of human chromosome 16. The results indicated that STP is localized proximal to the gene for protein kinase C, beta 1 polypeptide (PRKCB1), in the region from the distal portion of 16p11.2 to p12.1. The human STP gene maps near the locus for Batten disease (CLN3). Furthermore, we have determined by genotyping of murine interspecific backcross progeny that the homologous gene in mouse (Stp) localizes to the syntenic region of mouse chromosome 7 near the D7Mit8 (at 54 cM) and D7Bir1 markers.

Amino Acid Sequence↗
PubMed1993

Cloning and characterization of a mouse 3-methyladenine/7-methyl-guanine/3-methylguanine DNA glycosylase cDNA whose gene maps to chromosome 11.

In Escherichia coli, the repair of 3-methyladenine (3MeA) DNA lesions by DNA glycosylases prevents alkylation induced cell death. We described previously the isolation of a human 3MeA DNA glycosylase (AAG) cDNA that maps to chromosome 16 and hybridizes to specific genomic DNA fragments from a number of mammals, including mouse. As a first step in the generation of a 3MeA DNA glycosylase deficient mouse by homologous replacement in embryonic stem cells, we have cloned the mouse 3MeA DNA glycosylase cDNA. The cloned 1095 base pair cDNA contains a complete 333 amino acid open reading frame that predicts a 36.5 kDa protein and hybridizes to a 1.5 kb mRNA transcript. Mouse 3MeA DNA glycosylase (Aag) transcript levels vary by up to 21 fold among tissues, being highest in the testes and lowest in the heart. The Aag cDNA encodes a glycosylase able to release 3MeA, 7-methylguanine (7MeG) and 3-methylguanine (3MeG) from alkylated DNA. The expression of Aag in E. coli provides substantial resistance against killing by methylating agents, but, unlike its E. coli counterparts, the Aag glycosylase fails to protect against killing by ethylating and propylating agents. A 232 amino acid stretch of the predicted mouse protein shares extensive amino acid identity with rat (93%) and human (83%) 3MeA DNA glycosylases and we observe that all three mammalian glycosylases have a bipartite nuclear localization signal. The Aag gene maps to mouse chromosome 11, suggesting a segment of conserved synteny between mouse chromosome 11 and human chromosome 16, which bears the human 3MeA DNA glycosylase gene. Cloning the mouse 3MeA DNA glycosylase cDNA is a step toward understanding the role of this DNA repair enzyme in mammals.

Adenine↗
PubMed1993

Detection of Borrelia burgdorferi antigens in cerebrospinal fluid.

We examined CSF for Borrelia burgdorferi antigens using antigen-capture ELISA and Western (immuno) blot. Antigen-capture ELISA was positive in 38 of 77 (49%) CSF samples obtained from neurologic patients with presumed B burgdorferi infection, compared with one of 34 (3%) CSF samples obtained from other neurologic disease controls who came from a region endemic for Lyme disease. Western immunoblot was positive for B burgdorferi antigens in 12 of 22 (55%) CSF samples from the B burgdorferi infected groups, compared with none of 11 CSF samples from the control group. CSF antigen detection should prove helpful in evaluating patients for suspected neurologic Lyme disease.

Adolescent↗
PubMed1993

Interspecific somatic hybrid of Ichang papeda with Valencia orange.

Protoplasts from the leaf of Ichang papeda (Citrus ichangensis Swingle) were fused with the protoplasts of embryogenic suspension culture of Valencia orange (Citrus sinensis Osbeck) in vitro by polyethylene glycol (PEG)-induced fusion. The regenerated embryoids were malformed and were transferred onto shoot induction medium. The shoots were then grafted on 15-day-old seedlings of trifoliate orange in vitro. Chromosome counts of the young leaves showed that the parents were diploids, 2n = 2x = 18, and the regenerated plants were tetraploids, 2n = 4x = 36. Peroxidase and glutamate oxaloacetate transaminase isozyme analysis confirmed that these tetraploids were somatic hybrids. They have the bands of both parents. The hybrid plants grew vigorously after transplanted into soil. Leaf morphology of the hybrid was similar to that of sweet orange.

Cells, Cultured↗