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Z Xia

Publications and source records attributed to Z Xia.

At least 37 records · Page 2Linked to original sources
PubMed2001

Isolation and expression of the ecdysteroid-inducible angiotensin-converting enzyme-related gene in wing discs of Bombyx mori.

We isolated a clone encoding a putative angiotensin-converting enzyme-related gene from the wing disc cDNA library of the silkworm, Bombyx mori (refer to as BmAcer). The predicted open reading frame encoded 648 amino acids with about 50% identities with the Drosophila melanogaster angiotensin-converting enzyme Ance and Acer. Northern analysis identified a 2.2-kilobase mRNA which was abundant in wing discs two days after the beginning of wandering. An accumulation of the transcript was observed approximately 2 h after 20-hydroxyecdysone (20E) exposure in vitro and was blocked slightly by a protein synthetic inhibitor. These data suggest that the transcription of the BmAcer gene is directly 20E-inducible.

Amino Acid Sequence↗
PubMed2001

Quantitative (13)C NMR analysis of lignins with internal standards.

Novel protocols for acquiring quantitative (13)C NMR spectra of lignins have been developed using the internal reference compounds 1,3,5-trioxane and pentafluorobenzene. Trioxane offers a convenient internal standard for collecting inverse gated proton decoupled (13)C NMR spectra for lignins, whereas pentafluorobenzene can be used to provide information on the amount of methine carbon using the DEPT experiment. In each case, the internal reference compounds provide single, un-overlapped sharp signals in the middle of the spectral region, permitting facile integration. These integrals could be used to determine the amounts of different structural features of lignins, expressed in absolute units of millimoles per gram. The optimum parameters for these experiments were validated for a variety of spectrometer platforms, and standard errors were determined for different spectral areas using lignin model compounds and "standard" lignins. In addition, the data derived for the International Round Robin "standard" lignins showed good agreement with the data from quantitative (31)P NMR spectroscopy and published data, obtained by independent laboratories using independent methods of analysis.

Carbon Isotopes↗
PubMed2001

New spirostanol saponins from Chinese chives (Allium tuberosum).

Three new spirostanol saponins have been isolated from the seeds of Allium tuberosum. On the basis of acid hydrolysis and comprehensive spectroscopic analysis, their structures were established as tuberoside J, (25R)-5alpha-spirostan-2alpha,3beta,27-triol 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranoside; tuberoside K, (25R)-5alpha-spirostan-2alpha,3beta,27-triol 3-O-alpha-L-rhamnopyranosyl-(1-->2)-[alpha-L-rhamnopyranosyl-(1-->4)]-beta-D-glucopyranoside; and tuberoside L, 27-O-beta-D-glucopyranosyl-(25R)-5alpha-spirostan-2alpha,3beta,27-triol 3-O-alpha-D-rhamnopyranosyl-(1-->2)-[alpha-L-rhamnopyranosyl-(1-->4)]-beta-D-glucopyranoside.

Allium↗
PubMed2001

Blocking protein geranylgeranylation is essential for lovastatin-induced apoptosis of human acute myeloid leukemia cells.

Lovastatin is an inhibitor of the enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the major regulatory enzyme of the mevalonate pathway. We have previously reported that lovastatin induces a significant apoptotic response in human acute myeloid leukemia (AML) cells. To identify the critical biochemical mechanism(s) essential for lovastatin-induced apoptosis, add-back experiments were conducted to determine which downstream product(s) of the mevalonate pathway could suppress this apoptotic response. Apoptosis induced by lovastatin was abrogated by mevalonate (MVA) and geranylgeranyl pyrophosphate (GGPP), and was partially inhibited by farnesyl pyrophosphate (FPP). Other products of the mevalonate pathway including cholesterol, squalene, lanosterol, desmosterol, dolichol, dolichol phosphate, ubiquinone, and isopentenyladenine did not affect lovastatin-induced apoptosis in AML cells. Our results suggest that inhibiting geranylgeranylation of target proteins is the predominant mechanism of lovastatin-induced apoptosis in AML cells. In support of this hypothesis, the geranylgeranyl transferase inhibitor (GGTI-298) mimicked the effect of lovastatin, whereas the farnesyl transferase inhibitor (FTI-277) was much less effective at triggering apoptosis in AML cells. Inhibition of geranylgeranylation was monitored and associated with the apoptotic response induced by lovastatin and GGTI-298 in the AML cells. We conclude that blockage of the mevalonate pathway, particularly inhibition of protein geranylgeranylation holds a critical role in the mechanism of lovastatin-induced apoptosis in AML cells.

Acute Disease↗
PubMed2001

DNA elements of the type 1 adenylyl cyclase gene locus enhance reporter gene expression in neurons and pinealocytes.

The Ca2+-stimulated type 1 adenylyl cyclase (AC1) contributes to several forms of synaptic plasticity and is the only known neurospecific adenylyl cyclase. Furthermore, the protein and mRNA levels of AC1 undergo a circadian oscillation in the pineal gland, and AC1 may play a pivotal role in regulating nocturnal melatonin synthesis. To better understand the expression of AC1, we isolated mouse genomic DNA clones of AC1. The transcription and translation start regions of mouse AC1 share extensive homologies with the bovine counterpart. The upstream proximal region has potential binding sites for transcription factors, including the steroid receptor family, the E-box factors, and Sp1. A 280-bp fragment that contains the transcription start site directed reporter gene expression in cultured cortical neurons and pinealocytes functioning as a basal neuro- and pineal-directed promoter. Interestingly, pinealocyte expression of the reporter gene was inhibited by increases in cAMP. This cAMP sensitivity may explain why AC1 mRNA in the pineal is low at night when cAMP is elevated and high during the day when cAMP signals drop. An adjacent 330-bp fragment interacted specifically with nuclear factor(s) that we designate binary E-box factor (BEF). Methylation interference and DNase I footprinting identified the BEF-binding site sequence as 5'-CCAAGGTCACGTGGC-3'. When linked to the basal tissue-directed promoter, this 15-bp sequence further enhanced reporter expression in neurons and pinealocytes. We propose that this 15-bp sequence may contribute to increased expression of AC1 in neurons and pinealocytes relative to other cells.

Adenylyl Cyclases↗
PubMed2001

Osteogenic stem-cell characterization and development: potentials for cytotherapy.

BACKGROUND: Primitive progenitors of bone tissue exist postnatally and exhibit stem-cell characteristics, as shown by extensive renewal potential, and capacity to differentiate into all characteristic connective tissue types, including bone, cartilage, fat, fibrous tissue, muscle and hemopoietic stroma. METHODS: A wide variety of investigative techniques have been applied to characterize and assess differentiation of the normally non-cycling osteogenic stem cells. These include methods to assess in vitro and in vivo differentiation potentials, the production and use of Abs to identify surface markers, the expression of specific genes and, more recently, incorporation of marker genes (beta-galactosidase, green fluorescent protein) to study cell fate after implantation at tissue sites. RESULTS: Some antigenic cell-surface molecules reactive with MAbs generated by a number of laboratories have been identified. For cell-fate studies, retroviral insertion of beta-galactosidase or green fluorescent protein genes into human marrow stromal progenitors has been accomplished with high efficiency. The stromal cell phenotype and cellular functions in vitro are not significantly altered by these genetic modifications. In vivo transplantation in immunodeficient animals demonstrates migration and persistence of marrow stromal cells to skeletal and other tissue sites. DISCUSSION: None of the Abs generated against surface markers of early progenitors are absolutely lineage and cell-stage specific, but the respective Ags appear to participate in cell adhesion and cell-signalling mechanisms. These may be important in stem-cell activation and subsequent early osteogenic development. Studies of cell fate indicate feasibility for future uses in therapy of bone deficiency diseases and the potential for development of gene therapy procedures in these and other conditions.

Antigens, Surface↗
PubMed2001

Molecular and biochemical analysis of two beta-galactosidases from Bifidobacterium infantis HL96.

Two genes encoding beta-galactosidase isoenzymes, beta-galI and beta-galIII, from Bifidobacterium infantis HL96 were revealed on 3.6- and 2.4-kb DNA fragments, respectively, by nucleotide sequence analysis of the two fragments. beta-galI (3,069 bp) encodes a 1,022-amino-acid (aa) polypeptide with a predicted molecular mass of 113 kDa. A putative ribosome binding site and a promoter sequence were recognized at the 5' flanking region of beta-galI. Further upstream a partial sequence of an open reading frame revealed a putative lactose permease gene transcribing divergently from beta-galI. The beta-galIII gene (2,076 bp) encodes a 691-aa polypeptide with a calculated molecular mass of 76 kDa. A rho-independent transcription terminator-like sequence was found 25 bp downstream of the termination codon. The amino acid sequences of beta-GalI and beta-GalIII are homologous to those found in the LacZ and the LacG families, respectively. The acid-base, nucleophilic, and substrate recognition sites conserved in the LacZ family were found in beta-GalI, and a possible acid-base site proposed for the LacG family was located in beta-GalIII, which featured a glutamate at residue 160. The coding regions of the beta-galI and beta-galIII genes were each cloned downstream of a T7 promoter for overexpression in Escherichia coli. The molecular masses of the overexpressed proteins, as estimated by polyacrylamide gel electrophoresis on sodium dodecyl sulfate-polyacrylamide gels, agree with their predicted molecular weights. beta-GalI and beta-GalIII were specific for beta-D-anomer-linked galactoside substrates. Both are more active in response to ONPG (o-nitrophenyl-beta-D-galactopyranoside) than in response to lactose, particularly beta-GalIII. The galacto-oligosaccharide yield in the reaction catalyzed by beta-GalI at 37 degrees C in 20% (wt/vol) lactose solution was 130 mg/ml, which is more than six times higher than the maximum yield obtained with beta-GalIII. The structure of the major trisaccharide produced by beta-GalI catalysis was characterized as O-beta-D-galactopyranosyl-(1-3)-O-beta-D-galactopyranosyl-(1-4)-D-glucopyranose (3'-galactosyl-lactose).

Amino Acid Sequence↗
PubMed2001

Construction and analysis of mouse strains lacking the ubiquitin ligase UBR1 (E3alpha) of the N-end rule pathway.

The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. In the yeast Saccharomyces cerevisiae, the UBR1-encoded ubiquitin ligase (E3) of the N-end rule pathway mediates the targeting of substrate proteins in part through binding to their destabilizing N-terminal residues. The functions of the yeast N-end rule pathway include fidelity of chromosome segregation and the regulation of peptide import. Our previous work described the cloning of cDNA and a gene encoding the 200-kDa mouse UBR1 (E3alpha). Here we show that mouse UBR1, in the presence of a cognate mouse ubiquitin-conjugating (E2) enzyme, can rescue the N-end rule pathway in ubr1Delta S. cerevisiae. We also constructed UBR1(-/-) mouse strains that lacked the UBR1 protein. UBR1(-/-) mice were viable and fertile but weighed significantly less than congenic +/+ mice. The decreased mass of UBR1(-/-) mice stemmed at least in part from smaller amounts of the skeletal muscle and adipose tissues. The skeletal muscle of UBR1(-/-) mice apparently lacked the N-end rule pathway and exhibited abnormal regulation of fatty acid synthase upon starvation. By contrast, and despite the absence of the UBR1 protein, UBR1(-/-) fibroblasts contained the N-end rule pathway. Thus, UBR1(-/-) mice are mosaics in regard to the activity of this pathway, owing to differential expression of proteins that can substitute for the ubiquitin ligase UBR1 (E3alpha). We consider these UBR1-like proteins and discuss the functions of the mammalian N-end rule pathway.

Adipose Tissue↗
PubMed2001

[Modeling research about bioremediation of organic pollutants in soil-water-microbes system].

A new Theory, organic pollutant sequestration inside soil particles, was applied in the research in order to explain the persistence of residual chemicals in remediation sites. Based on this theory, a mathematical model which simulates organic pollutant bioremediation process in soil-water-microbes system was developed. In the model, diffusion is represented by Fick's second law, reversible sorption-desorption by a linear isotherm, irreversible sequestration by a pseudo-first order kinetics, and biodegradation by Monod kinetics. Model results match successfully with experimental data. Model simulations are performed in the study. It is noteworthy that the mathematical model will be useful in quantitatively predicting the time and degradation extend of organic pollutant in remediation sites.

Biodegradation, Environmental↗
PubMed2001

Cerivastatin triggers tumor-specific apoptosis with higher efficacy than lovastatin.

The statin family of drugs inhibits 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, the rate-limiting enzyme of the mevalonate pathway, and is used clinically as a safe and effective approach in the control of hypercholesterolemia. We have shown previously (Dimitroulakos, J., Nohynek, D., Backway, K. L., Hedley, D. W., Yeger, H., Freedman, M. H., Minden, M D., and Penn, L. Z. Increased sensitivity of acute myelogenous leukemias to lovastatin-induced apoptosis: a potential therapeutic approach. Blood, 93: 1308-1318, 1999) that lovastatin, a prototypic member of the statin family, can induce apoptosis of human acute myeloid leukemia (AML) cells in a sensitive and specific manner. In the present study, we evaluated the relative potency and mechanism of action of the newer synthetic statins, fluvastatin, atorvastatin, and cerivastatin, to trigger tumor-specific apoptosis. Cerivastatin is at least 10 times more potent than the other statins at inducing apoptosis in AML cell lines. Cerivastatin-induced apoptosis is reversible with the addition of the immediate product of the HMG-CoA reductase reaction, mevalonate, or with a distal product of the pathway, geranylgeranyl pyrophosphate. This suggests protein geranylgeranylation is an essential downstream component of the mevalonate pathway for cerivastatin similar to lovastatin-induced apoptosis. The enhanced potency of cerivastatin expands the number of AML patient samples as well as the types of malignancies, which respond to statin-induced apoptosis with acute sensitivity. Cells derived from acute lymphocytic leukemia are only weakly sensitive to lovastatin cytotoxicity but show robust response to cerivastatin. Importantly, cerivastatin is not cytotoxic to nontransformed human bone marrow progenitors. These results strongly support the further testing of cerivastatin as a novel anticancer therapeutic alone and in combination with other agents in vivo.

Acute Disease↗
PubMed2001

[The effects of ZMS on learning and memory ability and brain choline acetyltransferase in scopolamine-induced mouse model].

OBJECTIVE: To observe the effects of ZMS on learning and memory ability and on brain choline acetyltransferase activity. METHODS: A single intraperitoneal injection of scopolamine was used to establish a dementia mouse model. The learning and memory ability was detected by step-through and sted-down tests. And the choline acetyltransferase activity in brain was determined by 3H-acetyl-CoA incorporation analysis. RESULTS: ZMS could significantly reduce wrongness frequences, prolong incubation period of scopolamine-induced dementia mice in step-through and step-down tests (P < 0.01). ZMS could also significantly improve the activity of brain choline acetyltransferase (ChAT) (P < 0.01). CONCLUSION: ZMS could improve the learning and memory ability and the activity of brain ChAT in scopolamine-induced dementia mice. However, ZMS had no significant dose-response effect on improving the activity of brain ChAT, and the effect of Tacrine was similar to that of ZMS, it seemed unlikely that ZMS had a direct action on the ChAT. More probably, ZMS exerted its effect on ChAT activity and on learning and memory ability via elevating brain M receptor density.

Animals↗
PubMed2001

[Screening of aberrant genes of immunocytes in severely scalded rats by inhibition of subtractive hybridization].

OBJECTIVE: To explore the relationship between the postburn changes of gene expression of immunocytes and postburn immunological functional disturbance in severely scalded rats. METHODS: Immunocytes of female F344 rats with severe scalding were harvested and the mRNA was extracted for the inhibition of the expression of aberrant gene selected by subtractive hybridization in contrast to that before scalding. The aberrant genes were cloned and sequenced and compared to those in Genebank for homology. RESULTS: The harvested immunocytes were identified by LM to be monocytes and lymphocytes. Multiple strips located at 200 approximately 400 bp were obtained by inhibiting subtractive hybridization and were cloned into PGEM-T easy plasmid vector. Thus the genebank of aberrant expression was constructed. Twenty clones were sequenced randomly and partial obtained known genes were in accord with postburn systemic changes. Several novel genes were accepted and registered by Genebank. CONCLUSION: There exhibited change in the gene expression of peripheral immunocytes at early postburn stage in rats inflicted by severe burn.

Animals↗
PubMed2001

The preparation of rat heme oxygenase-1 mutant to reduce the level of bilirubin.

OBJECTIVE: To prepare rat heme oxygenase-1 (HO-1) mutants and to determine the activity and inhibition of this mutated enzyme. METHODS: pcDNA3HO1 containing truncated native rat HO-1 cDNA and pcDNA3HO1 delta 25 carrying mutated rat HO-1 cDNA (His25Ala) were constructed, respectively. COS-1 cells transfected with pcDNA3HO1 and pcDNA3HO1 delta 25 were collected and their activities were analyzed. RESULTS: Native rat HO-1 was highly expressed in transfected cells and its activity was 13,688-15,600 U/mg protein per hour. However, the enzyme activity of mutated HO-1 declined and the value was 1948-2160 U/mg protein per hour. When an equal amount of mutant was added to the enzyme reaction system, the level of bilirubin decreased by 42%. CONCLUSION: The His25Ala mutant reduced the formation of bilirubin, suggesting that the mutant could completely bind the heme with native enzyme.

Animals↗
PubMed2001

Alpha 1-antitrypsin and alpha 1-acid glycoprotein reduce the sensitivity of human dermal fibroblast to endotoxin.

OBJECTIVE: To test the hypothesis that acute phase reactants, such as alpha 1-antitrypsin and alpha 1-acid glycoprotein, could protect mammalian cells from further damage. METHODS: Human dermal fibroblasts (5 x 10(4)) were cultured with DMEM plus 10% FBS at 37 degrees C in a 5% CO(2) incubator. Different doses of LPS (lipo polysaccharide) and/or acute phase reactants were added. After 24 hours, the cultured supernatant was aspirated, the cells were washed, fixed and stained by methylene blue. The unbound stain was washed off. The stained cells were solubilize d in 0.1 ml of 1% Triton X-100. The absorbance of each well was measured us ing an ELISA spectrophotometer. The concentration of LPS which decreased the absorbance to 70% of the control (LPS-free) cultures was defined as LD(30). RESULTS: In order to achieve LD(30) in the presence of acute phase proteins, it was necessary to alter the LPS concentrations. The LD(3 0) of LPS treated with 0, 0.5, 2, 10 mg/ml antitrypsin and 0, 0.5, 2, 10 mg/ml glycoprotein was 5.4, 6.5, 7.6, 14.2 mg/ml and 5.2, 5.9, 6.9, 10.5 mg/ml, respectively. Statistically, with the treatment of more than 2 mg/ml antitrypsin or glycoprotein, LD(30) increased significantly. CONCLUSIONS: Our data show that fibroblasts are susceptible to the direct toxicity of LPS. Alpha 1-antitrypsin and alpha 1-acid glycoprotein can reduce the toxicity and/or increase the tolerance of mammalian cells to LP S.

Acute-Phase Reaction↗
PubMed2001

[A study on the myocardial contractile function and intracellular free calcium in scalded rats].

OBJECTIVE: To explore the relationship between postburn myocardial function and intracellular free calcium concentration. METHODS: Sprague-Dawley (SD) rats were employed as the model in which 7 were inflicted with 43% TBSA degree scalding (S group) and 7 as sham scalding control group (C group). The hearts of the injured rats were isolated and perfused in vitro by Langendorff method after 24 postburn hours (PBHs). The hearts were connected to a cardiac function monitor, and the dynamic changes in left ventricular develop pressure (LVDP) were continuously monitored. The cardiac 19F and 31P NMR spectroscopy was determined before and after and TF-BAPTA was added to the perfusate. RESULTS: When compared with those in C group, the LVDP of scalded rat heart decreased by 40% (P < 0.01), and the myocardial cytoplasmic free calcium concentration was four times above that in C group (P < 0.01). After TF-BAPTA was taken by myocytes, cardiac LVDP only decreased by 15% to 20%, and the PCr/Pi ratio decreased, while there was no change in ATP. CONCLUSION: Myocardial contractile function could be inhibited after burn injury, which might be related to the increase of myocardial cytoplasmic free calcium concentration.

Animals↗
PubMed2001

[Effect of transfected Cx43 gene on the gap junction intercellular communication and the human glioma cells proliferation].

OBJECTIVE: To investigate the role of connexin 43 (Cx43) gene in the gap junction intercellular communication (GJIC) and the development of glioma so as to demonstrate the feasibility of using Cx43cDNA as the target of gene therapy for gliomas. METHODS: TJ905 human glioblastoma cell line without Cx43 gene expression was transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunocytochemistry were used for examination of Cx43mRNA and its protein examination; MTT assay and silver colloid stain were used to detect cell proliferation. TUNEL method was used to determine the cell apoptosis. Scrape loading dye transfer (SLDT) was used to monitor GJIC. RESULTS: The TJ905 transfectants resulted in dramatic upregulation of Cx43mRNA and its protein, with the GJIC of transfected glioma cells restored. Clones with high Cx43 expression also showed reduced proliferation in vitro. However, the cell apoptosis did not increase following the transfection with Cx43 gene. CONCLUSION: These results indicate that Cx gene and gap junction intercellular communication are of crucial importance in malignant glioma cell growth and development. Cx43 gene may be a target for gene therapy of gliomas.

Blotting, Northern↗
PubMed2001

[Bioactivity and test grafting of acellular dermal matrix containing fibroblasts].

OBJECTIVE: To investigate the bioactivity of acellular dermal matrix with fibroblasts and its role as dermal skeleton. METHODS: Human fibroblasts (HFs) were planted onto the surface of acellular dermal matrix (ADM) to form living dermal substitute. The IL -- 6, IL -- 8 and TGF contents in the supernatant of the culture of HF -- ADM were determined with ELISA method, and the secretion of hyaluronic acid and laminin from extracellular matrix was measured with RIA method. The speed of vascularization and the wound contracture rate were observed after the dermal substitute was grafted on the full skin loss wound of Balb/c-nu mice (nude mice). RESULTS: HFs grew very well after being planted onto ADM so as to form a single layer of cellular membrane. Many kinds of cytokines and extra cellular matrix components were secreted. Compared with simple acellular dermal grafting, the vascularization was accelerated, and the wound contracture rate decreased, after the living dermal substitute being grafted on the wound. CONCLUSION: The ADM seeded with HFs exhibited excellent bioactivity and might be an optimal dermal substitute.

Animals↗