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Biomedical subjects

F Zhou

Publications and source records attributed to F Zhou.

At least 19 recordsLinked to original sources
PubMed2000

Distinctness of spore and vegetative cellular fatty acid profiles of some aerobic endospore-forming bacilli.

A gas chromatographic analysis method was employed to determine the cellular fatty acid (CFA) profiles of spores and vegetative cells of some aerobic endospore-forming bacilli. The harvests of experimental strains were processed to obtain pure spores and acquire whole cell fatty acid methyl esters for the subsequent gas chromatographic analysis, and the corresponding vegetative cells were set as control. Evaluation of reproducibility of spore CFA components revealed that, provided under standardized experimental procedure, spore CFA composition was stable enough for research purposes. Fatty acids recovered in spores in greater quantities were saturated branched-chain acids containing 15 and 17 carbon atoms, similar to the vegetative cells. Commonly, the proportions of saturated branched-chain acids in spores were greater than in vegetative cells. The dendrograms obtained by cluster analysis provided some meaningful taxonomic information of the experimental strains. The fatty acids analysis of spores seems to be a promising supplementary tool for the chemotaxonomic research of aerobic endospore-forming bacilli.

Bacillaceae↗
PubMed1999

A novel class of eukaryotic zinc-binding proteins is required for disease resistance signaling in barley and development in C. elegans.

Barley Rar1 is a convergence point in the signaling of resistance to powdery mildew, triggered by multiple race-specific resistance (R) genes. Rar1 is shown to function upstream of H2O2 accumulation in attacked host cells, which precedes localized host cell death. We isolated Rar1 by map-based cloning. The sequence of the deduced 25.5 kDa protein reveals two copies of a 60-amino acid domain, CHORD, conserved in tandem organization in protozoa, plants, and metazoa. CHORD defines a novel eukaryotic Zn2+-binding domain. Silencing of the C. elegans CHORD-containing gene, chp, results in semisterility and embryo lethality, suggesting an essential function of the wild-type gene in nematode development. Our findings indicate that plant R genes have recruited a fundamental cellular control element for signaling of disease resistance and cell death.

Alleles↗
PubMed1999

Apoptosis induced by oxysterol in CEM cells is associated with negative regulation of c-myc.

Previously we have demonstrated that treatment of the human lymphoblastic leukemic CEM cells with 25-hydroxycholesterol (25OHC) induces apoptosis. In the present study, we show that both c-myc mRNA and c-Myc protein levels are reduced only in oxysterol-sensitive and not in oxysterol-resistant cells after treatment with concentrations of 25OHC that kill the sensitive CEM cells. The repression of c-Myc protein precedes c-myc mRNA reduction, and both events occur before the onset of cell death. Our data suggest that 25OHC-induced suppression of c-myc gene expression in CEM cells results from posttranscriptional regulation. These results demonstrate the regulation by an oxysterol of a gene/gene product important for cell growth and viability and an association between oxysterol-induced apoptosis of CEM cells and the negative regulation of c-myc.

Apoptosis↗
PubMed1999

Increased injury following intermittent fetal hypoxia-reoxygenation is associated with increased free radical production in fetal rabbit brain.

Hypoxia associated with perinatal events can result in brain damage in the neonate. In labor and eclampsia, hypoxia can be intermittent, which may result in more severe damage than sustained hypoxia. The pathogenesis of brain injury in sustained ischemia involves free radical production; therefore, we investigated whether higher levels of free radicals contribute to the greater injury induced by repetitive ischemia. Brains were obtained from fetuses of near-term, pregnant rabbits subjected to repetitive ischemia-reperfusion (RIR), sustained uterine ischemia-reperfusion (IR), or a control protocol. Compared with controls, fetal brains from RIR or IR groups had more brain edema. Brains from RIR fetuses exhibited higher levels of lipid peroxidation, 3-nitrotyrosine, and nitrogen oxides, and lower total antioxidant capacity and cortical cellular viability than those of IR or control fetuses. Maternal administration of antioxidants following RIR and fetal bradycardia resulted in lower levels of fetal cortical and hippocampal cell death. Coadministration of Trolox and ascorbic acid resulted in less brain edema and liquefaction, and fewer hippocampal ischemic nuclei as compared with the saline control. Higher free radical production may be responsible for the greater fetal brain injury following repetitive hypoxia-reoxygenation. Maternal antioxidant treatment resulted in transplacental passage of antioxidants and amelioration of brain injury, and may be a viable clinical option following diagnosis of fetal distress.

Animals↗
PubMed1998

Tetranucleotide GGGA motif in primary RNA transcripts. Novel target site for antisense design.

Selecting effective antisense target sites on a given mRNA molecule constitutes a major problem in antisense therapeutics. By trial-and-error, only 1 in 18 (6%) of antisense oligonucleotides designed to target the primary RNA transcript of tumor necrosis factor-alpha (TNF-alpha) strongly inhibited TNF-alpha synthesis. Subsequent studies showed that the area in RNA targeted by antisense oligonucleotides could be moved effectively 10-15 bases in either direction from the original area. We observed that only molecules that incorporated a tetranucleotide motif TCCC (complementary to GGGA on RNA) yielded potent antisense oligonucleotides against TNF-alpha. A comprehensive literature survey showed that this motif is unwittingly present in 48% of the most potent antisense oligonucleotides reported in the literature. This finding was prospectively used to predict the sequences of additional antisense oligonucleotides for the rat TNF-alpha primary RNA transcript. Over 50% of antisense constructs (13 of 22) containing the TCCC motif were found to effectively inhibit TNF-alpha synthesis. Marked reductions in mRNA were also observed. This motif was found to be most effective when targeting introns in the primary RNA transcript, suggesting a nuclear localization for the antisense action. Predicting target sites based on the presence of this motif in primary RNA transcripts should be of value in the development on new antisense pharmacotherapy.

Animals↗
PubMed1998

Hypoxia-reoxygenation is as damaging as ischemia-reperfusion in the rat liver.

OBJECTIVE: We hypothesized that the extent of injury and release of xanthine oxidase, an oxidant generator, into the circulation would be less in normal-flow hypoxia-reoxygenation than in equal duration no-flow ischemia-reperfusion. DESIGN: Randomized study. SETTING: University-based animal research facility. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: The livers were isolated, perfused, and then randomly subjected to 2 hrs of hypoxia (normal flow, low oxygen) or ischemia (no flow, no oxygen), and 2 hrs of reperfusion. Hepatocytes were also isolated, and were subjected to either: a) hypoxia (0, 2, 4, and 6 hrs); or b) hypoxia (2 and 4 hrs) with reoxygenation (2 hrs). MEASUREMENTS AND MAIN RESULTS: The extent of liver injury (as assessed by release of hepatocellular enzymes) and the release of xanthine oxidase were measured from isolated-perfused rat livers and cultured hepatocytes. The pattern of release of xanthine oxidase in isolated-perfused liver effluent was different in hypoxia-reoxygenation compared with ischemia-reperfusion. During hypoxia, xanthine oxidase gradually increased in the effluent; then, the xanthine oxidase decreased to low concentrations during reoxygenation. After ischemia, there was a sharp spike in xanthine oxidase at 1 min of reperfusion, with a rapid decrease to low concentrations. The total release of xanthine oxidase during hypoxia-reoxygenation was similar to that during ischemia-reperfusion. Lactate dehydrogenase and other markers of liver injury showed a pattern of release that was similar to that of xanthine oxidase, but the total release of markers was not different between the two groups. In hepatocytes, most of the release of enzymes occurred in hypoxia, and the rate of release was not different between hypoxia and hypoxia-reoxygenation. CONCLUSIONS: Hypoxia-reoxygenation results in as much damage to the liver as ischemia-reperfusion, and results in the release of a similar amount of oxidant-producing xanthine oxidase into the circulation.

Alanine Transaminase↗
PubMed1998

Sustained hypoxia-ischemia results in reactive nitrogen and oxygen species production and injury in the premature fetal rabbit brain.

Free radical-mediated injury is implicated in hypoxic-ischemic encephalopathy observed in neonates. We investigated in utero free radical production and injury following hypoxia-ischemia to premature fetal brain utilizing a rabbit model of acute placental insufficiency. Pregnant rabbits at 29 days gestation were randomized to uterine ischemia for 50 minutes (min) (hypoxia) or nonischemic controls. Fetal brains were obtained immediately after ischemia for oxidative and acute-injury markers or 24 hours (h) post-ischemia for histopathology. Nitrotyrosine formation, a marker of NO-derived species such as peroxynitrite, was observed only in hypoxic brains. Hypoxia resulted in a significant increase in nitrogen oxides, lipid peroxidation, and protein oxidation, with a concomitant decrease in total antioxidant capacity, compared with controls. Peroxynitrite addition to brain homogenate increased nitrogen oxides linearly (1:1), although protein carbonyls were unchanged. Concomitantly, in vitro cortical and hippocampal cell viability and ATP levels decreased, with an increase in brain edema in hypoxic brains. Fetuses delivered 24 h post-ischemia had increased hippocampal nuclear karyorrhexis on histology compared with controls. Antioxidant administration (ascorbic acid and Trolox) intraperitoneally ameliorated changes in cellular viability and brain edema. Acute fetal hypoxia-ischemia without reoxygenation results in increased nitrogen and oxygen free radical production that may cause brain injury. The merits of the described model are discussed.

Animals↗
PubMed1998

Molecular characterization of the oxalate oxidase involved in the response of barley to the powdery mildew fungus.

Previously we reported that oxalate oxidase activity increases in extracts of barley (Hordeum vulgare) leaves in response to the powdery mildew fungus (Blumeria [syn. Erysiphe] graminis f.sp. hordei) and proposed this as a source of H2O2 during plant-pathogen interactions. In this paper we show that the N terminus of the major pathogen-response oxalate oxidase has a high degree of sequence identity to previously characterized germin-like oxalate oxidases. Two cDNAs were isolated, pHvOxOa, which represents this major enzyme, and pHvOxOb', representing a closely related enzyme. Our data suggest the presence of only two oxalate oxidase genes in the barley genome, i.e. a gene encoding HvOxOa, which possibly exists in several copies, and a single-copy gene encoding HvOxOb. The use of 3' end gene-specific probes has allowed us to demonstrate that the HvOxOa transcript accumulates to 6 times the level of the HvOxOb transcript in response to the powdery mildew fungus. The transcripts were detected in both compatible and incompatible interactions with a similar accumulation pattern. The oxalate oxidase is found exclusively in the leaf mesophyll, where it is cell wall located. A model for a signal transduction pathway in which oxalate oxidase plays a central role is proposed for the regulation of the hypersensitive response.

Amino Acid Sequence↗
PubMed1998

Optimization of the processing of matrix pellets based on the combination of waxes and starch using experimental design.

An experimental design was used in order to optimize the one-step production process of matrix pellets based on the combination of waxes and starch. The parameters tested were the impeller speed (x1) and the mixing time (x2). Ibuprofen and theophylline were used as model drugs at a concentration of 60 and 70% (w/w), respectively. The 0.8-1.25 mm yield fraction of the matrix pellets was evaluated as the response factor Y. A quadratic equation was fitted to the experimental data and used to predict the response factor Y of the theophylline and the ibuprofen. The contour plots of both formulations revealed a flat and therefore rugged region from the upper left to the lower right of the domain investigated. The energy input into the system during the production process controlled the pellet growth, the impeller speed having a greater impact on the energy input compared to the mixing time.

Delayed-Action Preparations↗
PubMed1998

[Influences of sperm quality and quantity on fertilization, cleavage rates and quality of embryos in in-vitro fertilization].

OBJECTIVE: To examine the influences of the quality and quantity of seminal sperm and inseminational sperm on in-vitro fertilization (IVF) outcomes. METHODS: chi-square tests were used to analyze the impacts of both semen and inseminated sperm concentrations and percentages of different grade motile sperm on the fertilization rates in 481 matured and 273 immatured oocytes and cleavage rate percentage of embryo with low quality in embryos obtained from 92 IVF-embryo transfer cycles. RESULTS: The fertilization rates of both kinds of oocytes and cleavage rate of fertilizaed matured oocyte significantly decreased (P < 0.05) as the concentrations and percentages of grade a and b sperms in semen reduced. The rates of embryos with abnormal morphology were significantly dropped when the above sperm parameters increased (P < 0.05). Meanwhile, it was also found that the concentration and motility of inseminated sperm could also affect the above three IVF parameters significantly (P < 0.05). CONCLUSION: Highly active semen and inseminated sperm play important roles not only in fertilization, but also in cleavage and embryo development during IVF procedure.

Cleavage Stage, Ovum↗
PubMed1998

Identification of the cornea-specific keratin 12 promoter by in vivo particle-mediated gene transfer.

PURPOSE: Keratin 12 (K12) is a cornea epithelial cell-specific intermediate filament component. To provide a better understanding of its expression, it is necessary to identify and characterize the promoter of Krt1.12 gene. METHODS: The 2.5-kb DNA 5' to Krt1.12 gene was sequenced. Krt1.12 promoter-beta-gal DNA constructs were prepared and used in vivo to transfect rabbit corneas, conjunctivas, and skin by particle-mediated gene transfer (Gene Gun). In vitro, the DNA constructs were transfected into cultured T-antigen-transformed rabbit corneal epithelial (RCE-T) cells and human fibrosarcoma HT-1080 fibroblasts with lipofectamine. The promoter activity was assessed by measuring beta-gal (beta-galactosidase) activity using histochemical staining with 5-Bromo-4-chloro-3-indolyl-beta-D-galactoside and enzyme assay with o-nitrophenyl beta-D-galactopyranoside. RESULTS: There are four Pax-6 pair box binding elements found between -910 and -2000 bp 5'-flanking the transcription initiation site of the Krt1.12 gene. None of promoter constricts can be expressed by HT-1080 cells. Cotransfection of Pax-6 cDNA with K12 promoter-beta-gal constructs containing Pax-6 elements results in a fourfold increase of beta-gal activities in RCE-T cells but not HT-1080 fibroblasts. The data of in vivo transfection in the rabbit by Gene Gun indicate that reporter gene constructs containing 0.6-kb and longer DNA fragments 5'-flanking Krt1.12 gene are effectively expressed in corneal, but not conjunctival or epidermal epithelial cells. CONCLUSIONS: The particle-mediated gene transfer is a suitable technique for in vivo delivery of transgenes to corneal epithelial cells. The 2.5-kb DNA fragment 5'-flanking Krt1.12 contains corneal epithelial cell-specific regulatory cis-DNA elements. Pax-6 is a positive transcription factor essential for keratin 12 expression.

Animals↗
PubMed1997

A heparin-binding synthetic peptide of heparin/heparan sulfate-interacting protein modulates blood coagulation activities.

We have previously identified and characterized a heparin-binding cell surface protein (heparin/heparan sulfate-interacting protein, or HIP) present on epithelial and endothelial cells. A synthetic peptide mimicking a heparin-binding domain of HIP is now shown to bind a small subset of heparin molecules with high affinity and, therefore, presumably recognizes a specific structural motif in the heparin molecule. Further analyses revealed that the heparin molecules exhibiting a high affinity for the HIP peptide also show an extremely high affinity for antithrombin III (AT-III), a cofactor required for heparin's anticoagulant activity. The HIP peptide was shown to compete with AT-III for binding to heparin and to neutralize the anticoagulant activity of heparin in blood plasma assays. Furthermore, the heparin subfraction that binds to the HIP peptide with high affinity exhibits an extremely high anticoagulant activity. We conclude that although the HIP peptide shows no sequence similarity with AT-III, the two proteins recognize the same or similar structural motifs in heparin.

Amino Acid Sequence↗
PubMed1997

[Idiopathic rapidly progressive glomerulonephritis and small vessel vasculitis].

We analyzed the clinical and pathological characteristics of patients with idiopathic rapidly progressive glomerulonephritis (idiopathic RPGN), who had been hospitalized in recent three years. Five out of 8 patients with idiopathic RPGN were positive for anti-neutrophil cytoplasmic autoantibodies (ANCA), most of them showed pANCA by indirect immunofluorescence technique. Compared with those who were ANCA negative, ANCA positive patients had an elder onset age; most of them had fever, arthragia, myalgia, skin rash, and some patients also had deafness, eye and nasal disorders. ANCA positive patients had a positive CRP, elevated gammaglobulins and accelerated ESR (> 100 mm/1 h). Typical pathological findings for ANCA positive patients were focal segmental fibroid necrosis of the glomerular capillary tufts and diffuse crescents formation. After an intensive therapy, the renal functions of most ANCA positive patients were obviously improved and most of them became hemodialysis independent. The results suggested that idiopathic RPGN might be a renal limited form of small vessel vasculitis and the patients with idiopathic RPGN had a better prognosis. Early diagnosis and appropriate intensive therapy are critical for patients with idiopathic RPGN.

Adult↗
PubMed1997

[Detection of p53 gene mutations in hepatocellular carcinoma].

This study screened 32 cases of hepatocellular carcinoma (HCC) from Chengdu detecting HBV DNA. HBsAg and p53 mutations by using Southern blot hybridization, immunohistochemistry and polymerase chain reaction/restriction enzyme digest methods, respectively. The results revealed all the cases had been infected by HBV; the frequency of HBV DNA integration into HCC cell was 72%, and the positive staining for HBsAg 96.3%(26/27). Of the 32 cases, 8 showed nuclear staining of p53 protein (25%), no mutation of p53 gene at all in 27 nontumorours liver tissues was identified. The resluts suggest the inactivation of p53 function may play a significant role in the genesis of HBV-associated HCC; however, the largely negative p53 mutation results in the current study conversely indicate that hepatocarcinogenesis may even involve other comprehensive mechanisms. Further studies are worth doing to evaluate the possible contribution of HBV to the p53 mutation in HCC.

Adult↗
PubMed1997

Isolation and identification of berberine from cell cultures of Coptis chinensis.

Calli were formed by the young leaves of Coptis chinensis inoculated onto a 6,7-V solid medium containing 1.0-2.0 mg/L of 2,4-D and 0.1 mg/L of kinetin. The loose calli were chosen and transferred into a liquid medium and free cells and cell aggregates were obtained. Cell lines with a higher content of berberine were selected by irradiation of suspension culture and a plate-screening technique. After 35 subcultures, the selected cell lines were cultured in a larger quantity for the extraction of alkaloids. Yellow crystals were obtained from the extracts and identified as berberine by TCL, UV and IR absorption, and mass spectrography. They have the same molecular structure and anti-bacterium activities as the berberine obtained from natural plants.

Berberine↗
PubMed1997

Familial occurrence of hemangioblastoma of central nervous system.

OBJECTIVE: To study the familial occurrence of hemangioblastoma of the central nervous system and to search for an effective way to diagnose and manage the disease. METHODS: From 1974 to 1993, six patients with hemangioblastoma with family history underwent surgical treatment at our hospital. Computerized tomography (CT) or magnetic resonance imaging (MRI) technique was used for the diagnoses before operation. RESULTS: Each patient had more than two relatives with hemangioblastoma. Multiple lesions were found in two of the six familial hemangioblastoma cases. All the patients recovered well after operation. So far, no victims have been discovered in the third generation. CONCLUSIONS: The high occurrence rate of familial hemangioblastoma cases suggests an involvement of hereditary factors. We need a long follow-up study on the descendants of familial hemangioblastoma patients. MRI is of diagnostic value in determining the location and nature of hemangioblastoma in both brain and spinal cord. A thorough examination is necessary for hemangioblastoma patients, and microsurgery is important in removing the tumor.

Adult↗