Biomedical subjects

T Zhao

Publications and source records attributed to T Zhao.

At least 37 records · Page 2Linked to original sources

PubMed2001

Chemiluminescence flow sensor for berberine with immobilized reagents.

A sensitive chemiluminescence (CL) sensor for berberine combined with flow-injection (FI) technology is described. The analytical reagents involved in the CL reaction, including luminol and potassium ferricyanide, were both immobilized on an anion-exchange resin column. While a volume of sodium phosphate was passed through the column, the two CL reagents were eluted from the resin and then mixed with a berberine stream under alkaline conditions. By means of the fast oxidation reaction between berberine and potassium ferricyanide, potassium ferrocyanide was generated, which then inhibited the CL reaction of luminol and potassium ferricyanide. The decreased CL intensity was correlated with the berberine concentration in the range from 0.05 to 300 ng mL(-1 )with a relative standard deviation of less than 4.3%, and the limit of detection was 0.02 ng mL(-1) at a flow rate of 2.0 mL min(-1). It was shown that the flow sensor could greatly improve the selectivity and sensitivity for determination of berberine. The determination of analyte could be performed in 2 min, including sampling and washing and be stable for over 300 analyses. Some interference and additives in preparations were tested. Two herbal medicines commonly found in compound prescriptions, rutin and baicalin, were studied preliminarily. The method has been applied for the determination of berberine in pharmaceutical preparations successfully.

Berberine↗

PubMed2001

Improved typing procedure for the polymorphic single-copy RLA-DQA gene of the rabbit reveals a new allele.

The DQA gene of the rabbit major histocompatibility complex (MHC, RLA) is highly polymorphic and, in contrast to those reported for other mammalian species, is present as a single copy. These properties allow use of this gene in a method to type the class II locus of RLA by a combination of single-stranded conformational polymorphism (SSCP) and heteroduplex (HD) analysis. Familial segregation of RLA-DQA was shown and RLA class II types for rabbits of unknown pedigree were determined using migration patterns of amplified genomic DNA. Typing results were confirmed in experiments where unknown samples were mixed with products from rabbits of RLA types defined by sequence analysis. These analyses detected an RLA-DQA allele in addition to the five previously described; this new allele is designated RLA-DQA-F.

Alleles↗

PubMed2001

Overexpression of Pti5 in tomato potentiates pathogen-induced defense gene expression and enhances disease resistance to Pseudomonas syringae pv. tomato.

The tomato Pti5 gene encodes a pathogen-inducible ethylene response element-binding protein-like transcription factor that interacts with the disease resistance gene product Pto. Overexpression of Pti5 or Pti5-VP16, a translational fusion with a constitutive transcriptional activation domain, in tomato enhanced resistance to Pseudomonas syringae pv. tomato. Constitutive expression of Pti5 or Pti5-VP16 did not affect the basal level of pathogenesis-related gene expression, but it accelerated pathogen-induced expression of GluB and Catalase. The results demonstrate a positive role of Pti5 in defense gene regulation and disease resistance and suggest that a pathogen-activated posttranscriptional regulatory step is necessary for the pathogen induction of the defense gene expression.

DNA-Binding Proteins↗

PubMed2001

Experimental and field studies of Escherichia coli O157:H7 in white-tailed deer.

Studies were conducted to evaluate fecal shedding of Escherichia coli O157:H7 in a small group of inoculated deer, determine the prevalence of the bacterium in free-ranging white-tailed deer, and elucidate relationships between E. coli O157:H7 in wild deer and domestic cattle at the same site. Six young, white-tailed deer were orally administered 10(8) CFU of E. coli O157:H7. Inoculated deer were shedding E. coli O157:H7 by 1 day postinoculation (DPI) and continued to shed decreasing numbers of the bacteria throughout the 26-day trial. Horizontal transmission to an uninoculated deer was demonstrated. Although E. coli O157:H7 bacteria were recovered from the gastrointestinal tracts of deer necropsied from 4 to 26 DPI, attaching and effacing lesions were not apparent in any deer. Results are similar to those of inoculation studies in calves and sheep. In field studies, E. coli O157 was not detected in 310 fresh deer fecal samples collected from the ground. It was detected in feces, but not in meat, from 3 of 469 free-ranging deer in 1997. In 1998, E. coli O157 was not detected in 140 deer at the single positive site found in 1997; however, it was recovered from 13 of 305 dairy and beef cattle at the same location. Isolates of E. coli O157:H7 from deer and cattle at this site differed with respect to pulsed-field gel electrophoresis patterns and genes encoding Shiga toxins. The low overall prevalence of E. coli O157:H7 and the identification of only one site with positive deer suggest that wild deer are not a major reservoir of E. coli O157:H7 in the southeastern United States. However, there may be individual locations where deer sporadically harbor the bacterium, and venison should be handled with the same precautions recommended for beef, pork, and poultry.

Animals↗

PubMed2001

Iron-sulfur flavoprotein (Isf) from Methanosarcina thermophila is the prototype of a widely distributed family.

A total of 35 homologs of the iron-sulfur flavoprotein (Isf) from Methanosarcina thermophila were identified in databases. All three domains were represented, and multiple homologs were present in several species. An unusually compact cysteine motif ligating the 4Fe-4S cluster in Isf is conserved in all of the homologs except two, in which either an aspartate or a histidine has replaced the second cysteine in the motif. A phylogenetic analysis of Isf homologs identified four subgroups, two of which were supported by bootstrap data. Three homologs from metabolically and phylogenetically diverse species in the Bacteria and Archaea domains (Af3 from Archaeoglobus fulgidus, Cd1 from Clostridium difficile, and Mj2 from Methanococcus jannaschii) were overproduced in Escherichia coli. Each homolog purified as a homodimer, and the UV-visible absorption spectra were nearly identical to that of Isf. After reconstitution with iron, sulfide, and flavin mononucleotide (FMN) the homologs contained six to eight nonheme iron atoms and 1.6 to 1.7 FMN molecules per dimer, suggesting that two 4Fe-4S or 3Fe-4S clusters and two FMN cofactors were bound to each dimer, which is consistent with Isf data. Homologs Af3 and Mj2 were reduced by CO in reactions catalyzed by cell extract of acetate-grown M. thermophila, but Cd1 was not. Homologs Af3 and Mj2 were reduced by CO in reactions catalyzed by A. fulgidus and M. jannaschii cell extracts. Cell extract of Clostridium thermoaceticum catalyzed CO reduction of Cd1. Our database sequence analyses and biochemical characterizations indicate that Isf is the prototype of a family of iron-sulfur flavoproteins that occur in members of all three domains.

Amino Acid Motifs↗

PubMed2001

Mitogen-activated protein kinases mediate heat shock-induced delayed protection in mouse heart.

We determined the role of p38 mitogen-activated protein kinase (MAPK), 72-kDa heat shock protein (HSP72), and antioxidant enzymes in whole body heat stress (HS)-induced cardioprotection in mouse hearts. Adult male mice were treated with either HS or anesthesia only. At 0.5, 48, 72, or 120 h later, the hearts were subjected to 20 min of global ischemia and 30 min of reperfusion in Langendorff mode. A significant protection against ischemia-reperfusion injury was observed 48 h after HS as demonstrated by: 1) reduction in infarct size; 2) decrease in leakage of lactate dehydrogenase; and 3) enhanced postischemic ventricular contractile function. No such protection was observed at other post-HS time points. HS caused an ~25% increase in phosphorylated c-Jun NH2-terminal kinase (JNK) but not p38 MAPK in the heart during the first 2-h post-HS time period. Cardioprotection was abolished by the MAPK inhibitor SB-203580, which also partially suppressed the HS-induced JNK phosphorylation. The protective effect was associated with a two- to threefold increase in HSP72 protein accumulation, but not antioxidant enzyme activities (catalase and Cu/Zn and Mn SOD) in the myocardium. Although HSP72 levels remained high 72 h after HS, the cardioprotection had already disappeared. We conclude that HS induces a transient delayed cardioprotection at 48 h after thermal stress in mice which appears to be mediated via a MAPK-signaling pathway.

Animals↗

PubMed2001

Chlorine inactivation of Escherichia coli O157:H7 in water.

Six human isolates of Escherichia coli O157:H7 and E. coli (ATCC 11229) were used to determine the concentrations of free chlorine and exposure times required for inactivation. Free chlorine concentrations of 0.25, 0.5, 1.0, and 2.0 ppm at 23 degrees C were evaluated, with sampling times at 0, 0.5, 1.0, and 2.0 min. Results revealed that five of six E. coli O157:H7 isolates and the E. coli control strain were highly susceptible to chlorine, with >7 log10 CFU/ml reduction of each of these strains by 0.25 ppm free chlorine within 1 min. However, comparatively, one of the seven strains was unusually tolerant to chlorine at 23 degrees C for 1 min, with a 4-, 5.5-, 5.8-, and >5.8-log CFU/ml reduction at free chlorine concentrations (ppm) of 0.25, 0.5, 1.0, and 2.0. respectively. Based on these studies most isolates of E. coli O157:H7 have no unusual tolerance to chlorine; however, one strain was exceptional in being recovered after 1-min of exposure of 10(7) CFU/ml to 2.0 ppm of free chlorine. This isolate may be a useful reference strain for future studies on chlorine tolerance of E. coli O157:H7.

Chlorine↗

PubMed2001

Evaluation of universal preenrichment broth for growth of heat-injured pathogens.

Universal preenrichment broth (UPB) was developed to enable enrichment of injured foodborne pathogens of different genera simultaneously in lieu of having to undergo separate simultaneous enrichment cultures for subsequent detection or isolation of each pathogen. Enrichment conditions in UPB for growth of injured pathogens to populations that will enable pathogen detection by rapid immuno-based or polymerase chain reaction (PCR)-based assays have not been defined. Hence, studies were done to determine recovery and growth rates of heat-injured Escherichia coli O157:H7, Salmonella enterica ser. Typhimurium, Salmonella enterica ser. Enteritidis. and Listeria monocytogenes in UPB. Bacterial cells were heat injured in tryptic phosphate broth at 57.2 degrees C and inoculated at populations of ca. 0.17 to 63 injured cells per ml with raw ground beef, fresh chicken, lettuce, and environmental sponge samples. Enrichment cultures were sampled at 1, 2, 3, 4, 5, 6, and 24 h at 37 degrees C postinoculation, and pathogens were enumerated on appropriate selective media. Results revealed that recovery and growth of pathogens during the first 6 h of enrichment were not sufficient to ensure adequate numbers of bacteria (> 10(3) CFU/ ml) for detection by most immunoassays or PCR assays. Cells often required 3 to 4 h for recovery before growth was initiated. Salmonella Typhimurium, Salmonella Enteritidis, E. coli O157:H7, or L. monocytogenes cell populations in enrichment cultures with ground beef or lettuce at 6 h were 0.5 to 2.9 log10 CFU/ml. At 24 h of incubation, cell counts of enrichment samples for the three pathogens from all food and environmental sponge samples ranged from 4.0 to 8.3 log10 CFU/ml. Enrichment in UPB at 37 degrees C of foods or environmental sponge samples containing heat-injured cells of Salmonella Typhimurium, Salmonella Enteritidis, E. coli O157:H7, or L. monocytogenes reliably provides at 24 h of incubation-but not at 6 h-sufficient cell populations for detection by rapid immunoassay or PCR assay procedures that can detect at least 4 log10 CFU/ml. These results raise questions regarding the sensitivity of rapid detection methods that employ an abbreviated enrichment protocol of 6 h or less.

Animals↗

PubMed2001

[Methanol method for the preparation of template DNA from blood for polymerase chain reaction].

A methanol method for the preparation of template DNA for polymerase chain reaction (PCR) from peripheral blood specimens was established. A diameter of 3 mm blood spots on paper were punched and fixed in methanol for overnight. Deionized water was then added. After boiling and centrifuging, the supernatants reserved DNA. The blood blot of specimen is easy to be obtained and reserved in low cost. The experiment technique is high in efficiency.

DNA↗

PubMed2001

[Hypoxic preconditioning enhances hypoxic tolerance of hippocampal neurons and synaptic function of rat].

Hypoxic preconditioning with different simulated altitudes (3,000 m and 5,000 m) was performed on Wistar rats and the evoked population spikes were recorded from the hippocampal slices of these rats. The results showed that the appearance of hypoxic injury potential (HIP) and the disappearance of presynaptic volley (PV) were significantly delayed in response to acute lethal hypoxia. HIP and PV delay became more apparent when the hypoxic preconditioning altitude was increased from 3,000 m to 5,000 m. After reoxygenation, the recovery rate of PV in hypoxic preconditioning groups at 3,000 m and 5,000 m was apparently higher than that of control. The above results suggest that hypoxic preconditioning of animals in vivo increases hypoxic tolerance of hippocampal neurons.

Animals↗

PubMed2001

Effect of Fas and Fas ligand deficiency in resistance of C57BL/6 mice to HSV-1 keratitis and chorioretinitis.

PURPOSE: To investigate the effect of Fas and Fas ligand (FasL) deficiency on the development of herpes stromal keratitis and on the von Szily model of herpes retinitis in C57BL/6 mice, which are ordinarily resistant to development of both of these herpetic diseases. METHODS: Anterior chamber inoculation of the right eye of each mouse with various titers of HSV-1 (KOS strain) was performed. Both eyes of each mouse were enucleated on postinoculation day 15 and processed for histopathologic examination. HSV-1 was inoculated into one cornea of other mice, and the severity of stromal keratitis was scored. RESULTS: Contralateral destructive chorioretinitis developed in susceptible Balb/cByj mice (19/23); ipsilateral chorioretinitis did not occur (0/23). Stromal keratitis developed in susceptible C.AL-20 mice (15/16). None of the C57BL/6 (0/10 for keratitis or 0/20 for retinitis) developed inflammation. Neither did B6.SMN.C3H.gld (FasL deficient; 0/12 or 0/28) or B6.MRL.lpr (Fas deficient; 0/11 or 0/34) mice (keratitis or contralateral chorioretinitis). Minimal scattering of inflammatory cells in the contralateral retina but not destructive chorioretinitis was observed in two C57BL/6, three B6.SMN.C3H.gld, and five B6.MRL.lpr mice. Few inflammatory cells were also found in the ipsilateral vitreous and vitreoretinal interface (but not destructive chorioretinitis) of all C57BL/6, two gld, and three lpr mice. CONCLUSIONS: Immune dysregulation secondary to deficiency in Fas or FasL system does not influence the resistance of the C57BL/6 mice to develop herpes simplex keratitis or destructive herpes simplex chorioretinitis.

Animals↗

PubMed2001

[Vitreous surgery for proliferative maculopathy].

OBJECTIVE: To explore the indication of vitrectomy for the treatment of pre-macular proliferative disease, the surgical technique, results and the prevention and treatment of complications. METHODS: Forty-one eyes of 41 cases with different kinds of macular traction or macular pucker, including 12 cases of idiopathic epimacular membrane, 10 cases of macular hole complicated with epimacular membrane and 19 cases of macular pucker after reattachment surgery (no diabetic retinopathy or retinal detachment) were treated with vitreous surgery. The issue of this operation is to find out underlying fissures (an under-bridge) or weakly adherent sites between a membrane and the retina. Using flute-needle, intraocular pick, spade and vitreoretinal forceps, we can separate and peel the proliferative membrane and the posterior cortical vitreous from the retinal surface completely. RESULTS: In 41 cases, epimacular membrane was peeled successfully, and the normal anatomic structure of the macula was restored. Four cases had small amount of hemorrhage in the superficial layer of the retina in and after the operation, and all the other cases did not have any complications, such as retinal tear or detachment. Thirty-eight cases obtained higher visual acuity, and in 3 cases the visual acuity maintained at their pre-operative level. Metamorphopsia was significantly improved, even disappeared. The follow-up period was 3 months to 4 years. CONCLUSIONS: The therapeutic effects of vitreous surgery for treatment of various kinds of proliferative maculopathy are reliable. After the surgery, the visual acuity can be elevated, the symptoms can be ameliorated and the complications can be effectively controlled.

Adult↗

PubMed2001

[Gigantism with low serum level of growth hormone: a case report].

Gigantism with low or normal basal concentrations of growth hormone (GH) is a rare condition, possibly due to abnormal GH secretory patterns, enhanced tissue sensitivity to GH, or the existence of an unidentified growth promoting factor. Here we report an 11 year-old female case of gigantism with a normal pituitary gland. Her height was 181 cm, body weight 77 kg, and bone age 11.1 years. Her basal serum GH levels were lower than 1 ng/ml. The levels of T3, T4, FT3, FT4, TSH, E2, LH, FSH, PRL, PTC and ACTH were normal. Serum GH response to insulin-induced hypoglycemia or arginine stimulation tests was blunted. In this case, non-pulsatile GH secretion and enhanced tissue sensitivity to GH may induce hypersecretion of IGF-1 and the existence of an unidentified growth promoting factor or biologically active anti-GH receptor antibodies may cause clinical gigantism.

Child↗

PubMed2001

[The levels of angiotensin and aldosterone in gases of hyperthyroid Graves' disease with and without hypokalemic periodic paralysis].

OBJECTIVE: To investigate the relationships between the changes of plasma angiotensin I (AT I). Angiotensin II (AT II). serum aldosterone (Ald) levels and hypokalemic periodic paralysis (THPP) complicating hyperthyroid Graves' disease. METHODS: The levels of serum potassium, TT3, TT4, FT3, FT4, Ald and the plasma AT I, AT II and 24 hours urine potassium output were determined in 73 cases of hyperthroid Graves' disease, of which 43 were complicated by THPP and the other 30 were grouped as NTHPP cases. RESULTS: The mean level of serum potassium in THPP group was significantly lower than that in NTHPP group (P = 0.000), and the mean level of 24 hours urine potassium output in THPP group was significantly higher than that in NTHPP group(P = 0.000); The mean levels of plasma AT II and serum Ald in THPP group in upright position were significantly higher than those in NTHPP group (P = 0.008 and 0.013, respectively). The mean levels of 24 hours urine potassium output, plasma AT II and serum Ald in upright position were negatively correlated to serum potassium in both groups. The level of serum Ald in upright position was positively correlated to the level of 24 hours urine potassium out-put. The mean levels of plasma AT I and AT II in supine position were positively correlated to serum T3, and the level of plasma AT II in upright position was positively correlated to serum FT3. CONCLUSION: The level of 24 hours urine potassium output rises in THPP patients; the high levels of AT and Ald may contribute to the pathogenesis of the disease.

Adult↗

PubMed2001

[Research on the immunological cause of auricular pseudocyst].

OBJECTIVE: To elucidate the relationship between the auricular pseudocyst and the immunological function of patients. METHOD: The cyst fluid and blood sample had been detected for contents of IgG, IgA, IgM and complement C3 by radial immunodiffusion, immunocomplex (IC) contents by polyethylene glycol turbidimetry in 55 cases, anti-nuclear antibody (ANA) by immuno-fluorescent technique in 23 cases, extractable nuclear antibody (ENA) by immunotransfer technique in 24 cases. The frozen sections of cyst wall tissue of 24 cases had been detected for immunocomplex by immunoenzyme histochemistric method. The reactions were observed separately between the auricle tissue of healthy white rat, human embryo and cyst fluid, serum of patient and normal serum. RESULT: Contents of IgG, IgA, IgM and C3 in the cyst fluid were lower than the serum significantly (P < 0.01). No IC, ANA and ENA could been found both in serum and cyst fluid, but reactive immunocomplex was discovered on the auricle cartilagous tissue of patient, immunocomplex also be observed on human embryo aurical tissue when it reacted with cyst fluid, but complex hadn't be found when the auricle tissue of white rat and human embryo reacted with serum of patient and normal people. CONCLUSION: The reason of auricular pseudocyst may be related with the local autoimmune status of patient.

Adult↗

PubMed2000

Phosphorylation site mutations in heterochromatin protein 1 (HP1) reduce or eliminate silencing activity.

HP1 is an essential heterochromatin-associated protein in Drosophila. HP1 has dosage-dependent effects on the silencing of euchromatic genes that are mislocalized to heterochromatin and is required for the normal expression of at least two heterochromatic genes. HP1 is multiply phosphorylated in vivo, and HP1 hyperphosphorylation is correlated with heterochromatin assembly during development. The purpose of this study was to test whether HP1 phosphorylation modifies biological activity and biochemical properties of HP1. To determine sites of HP1 phosphorylation in vivo and whether phosphorylation affects any biochemical properties of HP1, we expressed Drosophila HP1 in lepidopteran cultured cells using a recombinant baculovirus vector. Phosphopeptides were identified by matrix-assisted laser desorption ionization/time of flight mass spectroscopy; these peptides contain target sites for casein kinase II, protein tyrosine kinase, and PIM-1 kinase. Purified HP1 from bacterial (unphosphorylated) and lepidopteran (phosphorylated) cells has similar secondary structure. Phosphorylation has no effect on HP1 self-association but alters the DNA binding properties of HP1, suggesting that phosphorylation could differentially regulate HP1-dependent interactions. Serine-to-alanine and serine-to-glutamate substitutions at consensus protein kinase motifs resulted in reduction or loss of silencing activity of mutant HP1 in transgenic flies. These results suggest that dynamic phosphorylation/dephosphorylation regulates HP1 activity in heterochromatic silencing.

Animals↗

PubMed2000

Heterochromatin protein 1 binds to nucleosomes and DNA in vitro.

Heterochromatin protein 1 (HP1) is a nonhistone chromosomal protein primarily associated with the pericentric heterochromatin and telomeres in Drosophila. The molecular mechanism by which HP1 specifically recognizes and binds to chromatin is unknown. The purpose of this study was to test whether HP1 can bind directly to nucleosomes. HP1 binds nucleosome core particles and naked DNA. HP1-DNA complex formation is length-dependent and cooperative but relatively sequence-independent. We show that histone H4 amino-terminal peptides bind to monomeric and dimeric HP1 in vitro. Acetylation of lysine residues had no significant effect on in vitro binding. The C-terminal chromo shadow domain of HP1 specifically binds H4 N-terminal peptide. Neither the chromo domain nor chromo shadow domain alone binds DNA; intact native HP1 is required for such interactions. Together, these observations suggest that HP1 may serve as a cross-linker in chromatin, linking nucleosomal DNA and nonhistone protein complexes to form higher order chromatin structures.

Acetylation↗

PubMed2000

Inducible nitric oxide synthase mediates delayed myocardial protection induced by activation of adenosine A(1) receptors: evidence from gene-knockout mice.

BACKGROUND: The mechanism of delayed preconditioning induced by activation of adenosine A(1) receptors (A(1)ARs) is not fully understood. We determined the role of inducible nitric oxide synthase (iNOS) in mediating adenosine-induced late cardioprotection using pharmacological inhibitors and iNOS gene-knockout mice. METHODS AND RESULTS: Adult male mice were treated with saline or an A(1)AR agonist, 2-chloro-N(6)-cyclopentyladenosine (CCPA). Twenty-four hours later, the hearts were perfused in Langendorff mode and subjected to 30 minutes of global ischemia followed by 30 minutes of reperfusion. 8-Cyclopentyl-1,3-dipropylxanthine (DPCPX; 0.1 mg/kg IP) and S-methylisothiourea (SMT; 3 mg/kg IP) were used to block A(1)ARs and iNOS, respectively. Infarct size (IS) was measured by triphenyltetrazolium chloride staining, and iNOS expression was measured by Western blots. Myocardial IS was reduced from 24.0+/-3. 2% in the saline group to 12.2+/-2.5% in CCPA-treated mice (P<0.05). The infarct-reducing effect of CCPA was abrogated by DPCPX (29.3+/-3. 4%) and SMT (32.3+/-2.6%) and was absent in mice with targeted ablation of iNOS (23.9+/-1.6%). CCPA produced improvement in postischemic end-diastolic pressure, developed pressure, and rate-pressure product, which was also blocked by DPCPX and SMT. Increased iNOS protein expression observed in CCPA-treated hearts was diminished by DPCPX. CONCLUSIONS: Selective activation of A(1)ARs produces delayed cardioprotection against ischemia/reperfusion injury in the mouse. Increased iNOS expression concomitant with the lack of protective effect of A(1)AR activation in iNOS gene-knockout mice suggests a direct cause-and-effect relationship of iNOS in adenosine-induced late cardioprotection.

Adenosine↗