Search PubMed⌕ Search

Biomedical subjects

Z Xia

Publications and source records attributed to Z Xia.

At least 91 records · Page 5Linked to original sources
PubMed1999

Effect of lidocaine on improving cerebral protection provided by retrograde cerebral perfusion: a neuropathologic study.

OBJECTIVE: To determine whether lidocaine can improve the neuropathologic results in canine brains after retrograde cerebral perfusion (RCP). DESIGN: Randomized, blinded, experimental study. SETTING: University animal laboratory. PARTICIPANTS: Mongrel dogs. INTERVENTIONS: Fourteen mongrel dogs were placed on 120 minutes of hypothermic (20 degrees C) RCP. Following the RPC, they then resumed cardiopulmonary bypass and rewarming for 60 minutes. In the lidocaine group (n = 8), lidocaine was administered continuously; in the control group (n = 6), normal saline was administered. Cerebral perfusion fixation was performed at the end of the experiment. MEASUREMENTS AND MAIN RESULTS: The number of ischemic cells in 200 neurons was counted in the parietal cortex, CA1 sector of the hippocampus, CA3 sector of the hippocampus, ventral posterolateral nucleus of the thalamus, and Purkinje cells of the cerebellar cortex. Those in the parietal cortex, CA1 sector of the hippocampus, and ventral posterolateral nucleus of the thalamus were significantly less in the lidocaine group than in the control group (25.8+/-17.3 v 53.7+/-12.0; p < 0.01; 17.0+/-8.5 v 54.7+/-22.1; p < 0.01; and 16.9+/-17.8 v 49.7+/-28.4; p < 0.05, respectively). The total number of ischemic cells in the five examined regions was also significantly less in the lidocaine group than in the control group (89.5+/-19.4 v 219.5+/-45.5; p < 0.01). CONCLUSION: Continuous lidocaine significantly alleviated the ischemic neuropathologic injury after RCP and thus possibly improved cerebral protection.

Anesthetics, Local↗
PubMed1999

Granzyme A loading induces rapid cytolysis and a novel form of DNA damage independently of caspase activation.

Cytotoxic lymphocytes trigger apoptosis by releasing perforin and granzymes (Grn). GrnB activates the caspase apoptotic pathway, but little is known about GrnA-induced cell death. Perforin was used to load recombinant GrnA and GrnB and enzymatically inactive variants into target cells. GrnA induces single-strand DNA breaks that can be labeled with Klenow polymerase and visualized on alkaline gels. GrnA-induced DNA damage but not cytolysis requires GrnA proteolysis. GrnA-induced membrane perturbation, nuclear condensation, and DNA damage are unimpaired by caspase blockade. GrnA fails to induce cleavage of caspase-3, lamin B, rho-GTPase, or PARP. GrnA-induced cytotoxicity and cleavage of PHAP II, a previously identified GrnA substrate, are unimpaired in Jurkat cells that overexpress bcl-2. Therefore, GrnA activates a novel apoptotic pathway.

Animals↗
PubMed1999

Potential effect of authorization bias on medical record research.

OBJECTIVE: To analyze the influence of recent changes in Minnesota statutes that generally require prior authorization for use of medical records for research from patients who received medical care after Jan. 1, 1997. MATERIAL AND METHODS: In this Mayo Clinic Institutional Review Board-approved study, we obtained a stratified random sample of patients encountered at Mayo Clinic Rochester during the period 1994 through 1996 and estimated the proportion willing to provide the general authorization. On the basis of data from administrative files, we then compared demographic, diagnostic, and utilization characteristics for patients who provided authorization and those who did not. RESULTS: Overall, 3.2% (95% confidence interval, 2.4 to 4.0%) of the study subjects declined authorization. If patients not responding to requests for authorization were also considered to have refused, the overall refusal rate would be 20.7% (95% confidence interval, 18.5 to 22.9%). Women were somewhat more likely to refuse authorization than were men (4.0% versus 2.4%; P = 0.067), and patients younger than 60 years were more likely to refuse than were older patients (5.4% versus 1.2%; P<0.001). Patients residing more than 120 miles from Rochester were much less likely to decline authorization than were local residents (2.1% versus 5.8%; P = 0.001). Patients with prior diagnoses that might be considered more sensitive such as mental disorders, infectious diseases, and reproductive problems also were more likely to refuse authorization. CONCLUSION: These data demonstrate that laws requiring written authorization for research use of the medical record could result in substantial biases in etiologic and outcome studies, the direction and magnitude of which may vary from topic to topic. Clinicians should be prepared to enter the discussion to help inform patients and legislators of the potential hazards of laws that restrict access to medical records for research purposes.

Adult↗
PubMed1999

[Study on antifungal mechanism of alpha-pinene].

The antifungal mechanism of alpha-pinene was studied using electronic microscopy and incorporation of isotope-labelled precursor. The results demonstrated that alpha-pinene had significant roles in inhibiting and killing Candida albicans. After treatment with alpha-pinene the fungal morphology and ultrastructure showed obvious changes: their cell wall and cytoplasmic membrane ruptured; intracellular components released out and the cell residue fused to form irregular masses. In addition, the synthesis of DNA, RNA, polysaccharide of cell wall and ergosterol of cytoplasmic membrane was inhibited. It is indicated that these changes are related to antifungal mechanism of alpha-pinene.

Antifungal Agents↗
PubMed1998

Expression of signal transducers and activators of transcription proteins in acute myeloid leukemia blasts.

Hematopoietic cytokine receptor signaling pathways involve activation of signal transducers and activators of transcription (STAT) proteins, which are postulated to be involved in cellular differentiation. Aberrant STAT isoforms (beta forms rather than the normal alpha forms) have been described and have been found to block the normal signaling pathway from the receptor. Bcr/Abl proteins have been suggested to directly activate STATs, without exposure to growth factors. We asked whether STATs play a role in leukemogenesis. We analyzed constitutive and induced patterns of STAT activity in pretreatment blasts from 36 newly diagnosed acute myeloid leukemia (AML) patients and studied protein tyrosine kinases (PTKs) that may be involved in STAT activity, using in vitro and in-gel kinase assays. The beta forms were expressed in 21 of 27 samples (78%). Constitutive STAT3 and STAT5 activity was found in samples from 28 and 22% of patients, respectively. Response to exogenous cytokines identified two groups. STAT activity in one group was modulated by exogenous cytokines: constitutive STAT activity increased in some patients but decreased or disappeared in response to cytokines in others. The second group was cytokine insensitive. Additionally, we found constitutive PTK activity in two patients whose blasts demonstrated constitutive STAT activity, suggesting that PTKs use cytokine receptor signal pathways to activate STATs in AML blasts without exposure to exogenous cytokines. Our data suggest that (a) constitutive expression of aberrant STATs may be involved in blocking differentiation of AML blasts, (b) exogenous cytokines may activate STAT-inhibitory pathways, and (c) STATs may be activated by PTKs in some AML blasts.

Acute Disease↗
PubMed1998

Structure and properties in solution of PsaD, an extrinsic polypeptide of photosystem I.

PsaD is a small, extrinsic polypeptide located on the stromal side (cytoplasmic side in cyanobacteria) of the photosystem I reaction centre complex. The gene from the cyanobacterium Nostoc sp. PCC 8009 was expressed in Escherichia coli and the structure of the recovered protein in solution investigated. Size-exclusion chromatography, dynamic light scattering and measurement of 15N transverse relaxation times showed that the protein is a stable dimer in solution, whereas in the reaction centre complex it is a monomer. NMR experiments showed that the dimer is symmetrical and that there are at least two domains, one structured and the remainder unstructured. The structured domain contains a small amount of beta-sheet. Three-dimensional heteronuclear NMR spectra of [13C, 15N]PsaD showed that the structured domain is associated with the central part of the sequence while the N- and C-terminal regions are mobile. Evidence was obtained for a shift in equilibrium between two slightly different conformational states at about pH 6, and the protein was shown to bind to PsaE preferentially at neutral pH. Addition of trifluoroethanol was shown to induce the formation of a small amount of alpha-helix, and the form present in 30% trifluoroethanol appears to be more closely related to the in situ structure, which has been reported to contain one short helix in crystals [Schubert, W.-D., Klukas, O., Krauss, N., Saenger, W., Fromme, P. & Witt, H. T. (1997) J. Mol. Biol. 272, 741-769]. The significance of these findings for the assembly of the complex is discussed.

Amino Acid Sequence↗
PubMed1998

Expression and purification of enzymatically active recombinant granzyme B in a baculovirus system.

Granzyme B (GranB), a serine protease stored in the granules of cytotoxic T lymphocytes and natural killer cells, can initiate target cell apoptosis. To produce large amounts of purified active enzyme, recombinant murine granzyme B (rGranB) was expressed from baculovirus in insect cells. The expressed rGranB is secreted into the culture medium and can be readily purified to homogeneity by one-step affinity chromatography to yield 1.5 mg enzyme per liter insect cell medium. RGranB is recognized by a GranB-specific anti-peptide antibody and is active against synthetic substrate Boc-Ala-Ala-Asp-SBzl with kinetic constant (kcat/Km 45,000 M-1s-1) comparable to purified human GranB, RGranB processes the caspase pro-CPP32 into its enzymatically active form and induces DNA fragmentation in isolated nuclei in the presence of cytosolic factors. The ability to express enzymatically active rGranB using the baculovirus system will help elucidate the role of this granzyme in the immune response.

Animals↗
PubMed1998

Modulation of apoptosis induced by tricyclic antidepressants in human peripheral lymphocytes.

We have recently reported that the tricyclic antidepressants (TCAs) imipramine, clomipramine, and citalopram induce apoptosis in human peripheral lymphocytes. This system is well suited for studies on the pathophysiology/physiology of apoptosis regulation. Apoptosis was determined using both DNA gel electrophoresis and flow cytometric analysis. TCA-induced apoptosis in lymphocytes was monitored in the presence of the protein synthesis inhibitor cycloheximide (CHX), the RNA synthesis inhibitor actinomycin D (Act D), the antioxidant reduced glutathione (GSH), the nuclease inhibitor aurintricarboxylic acid (ATA), the cytokine interleukin-2 (IL-2), and the immunostimulator linomide. CHX and Act D failed to prevent and actually enhanced TCA-induced apoptosis in lymphocytes, indicating that protein and RNA syntheses are not required for this process. Exogenous IL-2, GSH, and ATA protected the lymphocytes from apoptosis induced by TCAs in a dose-dependent manner, whereas linomide had no effect on TCA-induced apoptosis under our in vitro conditions. Our data demonstrate that TCA-induced apoptosis in human lymphocytes shares many common features with other stimuli-induced apoptotic processes.

Antidepressive Agents, Tricyclic↗
PubMed1998

Recovering and reamplifying of the differentially expressed cDNA bands isolated from mRNA differential display. A modified method.

Methods for retrieving and reamplifying the differentially expressed cDNA bands have been modified. Direct reamplification of differentially expressed bands after cutting from a polyacrylamide gel (PAG) followed by a simple rinse and crush step has proved to be more convenient and effective than the traditional glycogen-precipitation method. Combination of 30 cycles of differential display (DD) polymerase chain reaction (PCR) and 20 cycles of standard PCR reaction also yielded higher reamplification rates.

DNA, Complementary↗
PubMed1998

Biodegradation of tricalcium phosphate ceramics by osteoclasts.

Biodegradation of tricalcium phosphate (TCP) ceramics was observed through mixed culture of osteoclasts and TCP discs in vitro in this study. Osteoclasts were isolated from newborn SD rat's marrow of long bone and cultured on TCP discs. The culture terminated at the 48th h and 96th h respectively. Under an inverted microscope, the osteoclasts imparted round or oval body with multinuclear and many thin processes. These cells were positively stained for tartrate-resistance acid phosphatase (TRAP). Scanning electron microscope showed that many resorption lacunae on TCP disc surface and their diameters were smaller than 20 microns. Osteoclasts were located in the lacunae. At the 96th h, the resorption lacunae become larger and osteoclasts showed degeneration. It is suggested that osteoclasts possess ability to re-absorb TCP ceramics under in vitro culturing condition.

Animals↗
PubMed1998

Development and characterization of monoclonal antibody to T-antigen: (gal beta1-3GalNAc-alpha-O).

The saccharide antigen, Gal beta1-3GalNAc or T antigen, is of biologic importance in many systems. It is a tumor-associated carbohydrate antigen, a temporally expressed antigen in germinal center B cells and cortical T cells, a parasite-associated antigen, a spermatozoa vitality marker and an antigen on aged red blood cells. It may play a role in normal cellular adhesion and in tumor cell metastasis. Well characterized monoclonal antibodies (MAb) to Gal beta1-3GalNAc will be useful for further studies in these areas. We developed an IgG3 MAb to Gal beta1-3GalNAc by immunizing the mice with a synthetic Gal beta1-3GalNAc-BSA conjugate. The MAb was analyzed using inhibition enzyme immunoassays with related synthetically prepared sugars to determine the restrictions involved in the antibody binding. Use of synthetic sugars as competitors enabled us to delineate the epitope restrictions on the binding activity of this monoclonal and will enable use of this MAb in studies concerning the biologic importance of this disaccharide.

Adenocarcinoma↗
PubMed1998

Nerve growth factor activates extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways to stimulate CREB serine 133 phosphorylation.

The mechanisms by which growth factor-induced signals are propagated to the nucleus, leading to the activation of the transcription factor CREB, have been characterized. Nerve growth factor (NGF) was found to activate multiple signaling pathways that mediate the phosphorylation of CREB at the critical regulatory site, serine 133 (Ser-133). NGF activates the extracellular signal-regulated kinase (ERK) mitogen-activated protein kinases (MAPKs), which in turn activate the pp90 ribosomal S6 kinase (RSK) family of Ser/Thr kinases, all three members of which were found to catalyze CREB Ser-133 phosphorylation in vitro and in vivo. In addition to the ERK/RSK pathway, we found that NGF activated the p38 MAPK and its downstream effector, MAPK-activated protein kinase 2 (MAPKAP kinase 2), resulting in phosphorylation of CREB at Ser-133. Inhibition of either the ERK/RSK or the p38/MAPKAP kinase 2 pathway only partially blocked NGF-induced CREB Ser-133 phosphorylation, suggesting that either pathway alone is sufficient for coupling the NGF signal to CREB activation. However, inhibition of both the ERK/RSK and the p38/MAPKAP kinase 2 pathways completely abolished NGF-induced CREB Ser-133 phosphorylation. These findings indicate that NGF activates two distinct MAPK pathways, both of which contribute to the phosphorylation of the transcription factor CREB and the activation of immediate-early genes.

Animals↗
PubMed1998

Microencapsulation of rat islets prolongs xenograft survival in diabetic mice.

OBJECTIVE: To protect the transplanted islets from the host's immune system by means of immunoexclusion membranes. METHODS: Rat islets were isolated from Wistar rat pancreas by ductal collagenase distention, stationary digestion, and finally with the aid of dextran gradient separation. Then the islets were encapsulated in alginate-polylysine-alginate (APA) semipermeable membranes. RESULTS: In vitro studies demonstrated that encapsulated islets secreted insulin in response to glucose challenge for at least 8 weeks, which was similar to free islets. In vivo studies showed that 15 streptozotocin (STZ)-induced diabetic mice were transplanted intraperitoneally with 1000 encapsulated islets without immunosuppression. Diabetes was reversed within 3 days, and the mice remained normoglycemic for up to 160 days, with a mean xenograft survival time of 126 days. The encapsulated islets had a significantly greater effect than unencapsulated islets, which functioned for less than 8 days. CONCLUSIONS: Encapsulation of pancreatic islets in semipermeable membranes can effectively prolong xenograft survival without immunosuppression in an animal model.

Animals↗
PubMed1998

[Detection of human cytomegalovirus by PCR in patients with gastrointestinal disease].

Human cytomegalovirus (HCMV) genomes was detected by PCR in 51 patients' biopsy or operation specimens with gastrointestinal disease. The positive rate was 9.80%. At the same time, 41 patients' serum HCMV-IgM was detected by indirect immunofluorescence assay (IFA); the positive rate was 46.34%. Patients with cancer and colitis exhibited a higher HCMV infection rate than those without cancer and colitis. The results suggest that PCR is sensitive and specific in detection of gastrointestinal HCMV infection, and the examination of biopsy or surgical specimen is superior than that of IFA of serum HMCV-IgG.

Adult↗
PubMed1998

The purification and identification of heme oxygenase isoforms from spleen tissue of rat and the expression of heme oxygenase-1 cDNA in COS-1 cells.

OBJECTIVE: To demonstrate that heme oxygenase (HO) isoforms exist in rat spleen treated with hematin and phenylhydrazine and to confirm that the isolated cDNA actually encodes HO-1 by expressing cDNA in monkey kidney cells (COS-1 cells) in order to prepare HO-1 mutant for inhibiting the natural enzyme. METHODS: The rat spleen microsomal fractions were first purified by diethylaminoethyl (DEAE)-Sephacel and hydroxylapatite. The activity of two isoforms (HO-1 and HO-2) of enzyme and their apparent molecular weight on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (SDS-PAGE) were measured. Secondly, using the isolated HO-1 cDNA clone, the expression plasmid pcDNA3HO1 was constructed and transfected into cultured COS-1 cells. The transfected cells were collected and disrupted by sonication, and the microsomes were prepared by ultracentrifugation. The activity of HO-1 was measured. RESULTS: Two isoforms were purified and identified in treated rat spleen and HO-1 was the predominant form. The ratio of HO-1 to HO-2 was 3.2:1. The apparent molecular weights of HO-1 and HO-2 were about 30 kD and 36 kD under reducing conditions, respectively. The HO-1 was highly expressed in endoplasmic reticulum of transfected cells. The specific band was located in molecular weight of 30 kD. The specific activity was five times higher than that of the control. CONCLUSION: The activity of expressed HO-1 in COS-1 cells is higher than that of purified enzyme from rat spleen tissue. It is suggested that this clone having an insert of 1030 base-pairs encodes HO-1 and that we can prepare HO-1 mutant by site-directed mutagenesis of HO-1 cDNA to prevent and treat hyperbilirubinemia.

Animals↗
PubMed1998

[DPOAE in high-risk neonatal screening for hearing].

64 neonates under intensive care have been tested by distortion product otoacoustic emission (DPOAE) and the auditory brain stem response (ABR) to investigate the value of DPOAE as a test for hearing impairment. The DPOAE test has been found to be practical and quick to perform. The proportion of NICU infants producing a recordable DPOAE is 83.6%, and the sensitivity and specificity to the ABR result is 92.4% and 92.2%, false positive is 7.8%, false negative is 7.6% respectively. These figures are high enough and the reduction in time compared to ABR is sufficient for the DPOAE to be considered as the primary screen, firm conclusions on the sensitivity of DPOAE to long term hearing impairment await the results from larger numbers of infants and further follow up data.

Deafness↗
PubMed1998

Lidocaine improving the cerebral protection by retrograde cerebral perfusion.

OBJECTIVE: To determine whether lidocaine can improve the cerebral protection provided by retrograde cerebral perfusion. METHODS: Sixteen mongrel dogs were placed on cardiopulmonary bypass and cooled to 20 degrees C. Retrograde cerebral perfusion was then carried out for 120 minutes, with the external jugular venous pressure kept at 3.33 kPa. Cardiopulmonary bypass was resumed, and the animals were rewarmed to 36 degrees C. The animals were divided into two groups. In the lidocaine group (n = 8), lidocaine was administrated continuously throughout the experiment. In the control group (n = 8), normal saline was given at the same rate. RESULTS: In both groups, cerebral tissue creatine phosphate and adenosine triphosphate concentrations and energy charge increased by the end of hypothermic cardiopulmonary bypass, decreased continuously during retrograde cerebral perfusion, and recovered gradually after resuming cardiopulmonary bypass. Nevertheless, they recovered to significantly higher levels in the lidocaine group than in the control group (creatine phosphate: 2.44 +/- 0.53 versus 1.61 +/- 0.49 mumol/g wet weight, P = 0.006; adenosine triphosphate: 0.71 +/- 0.18 versus 0.50 +/- 0.17 mumol/g wet weight, P = 0.029; energy charge: 0.59 +/- 0.10 versus 0.48 +/- 0.09, P = 0.044) by the end of the experiment. There was no significant difference between the two groups in the cerebral tissue water content (control group: 77.6% +/- 1.9%; lidocaine group: 77.6% +/- 1.3%). CONCLUSION: Continuous lidocaine infusion accelerates the recovery of cerebral tissue high energy phosphate contents after resuming cardiopulmonary bypass, but it has no effect on the formation of cerebral edema after retrograde cerebral perfusion.

Adenosine Diphosphate↗