Search PubMed⌕ Search

Biomedical subjects

Z Shao

Publications and source records attributed to Z Shao.

At least 127 records · Page 7Linked to original sources

[HLA-DR-positive T lymphocytes in the bone marrow and peripheral blood of patients with severe aplastic anemia and their in vitro inhibitory effecton BFU-E and CFU-GM].

OBJECTIVE: [corrected] To elucidate the role of immunodysfunction in the pathogenesis of severe aplastic anemia (SAA). METHODS: HLA-DR+ T cells were detected in bone marrow (BM) and peripheral blood (PB) of 20 SAA patients and controls (9 for BM and 11 for PB). PHA-LYCM conditioned medium was prepared, and its in vitro effect on normal bone marrow BFU-E and CFU-GM was studied. RESULTS: The percentages of HLA-DR+ T lymphocytes in BM and PB of newly diagnosed SAA patients were significantly higher than that of controls (P<0.001), and that of recovered SAA (rSAA) patients after ALG therapy (P<0.05). Compared with controls, the PBA-LYCMs from newly diagnosed SAA patients showed significant inhibitory effect on normal BFU-E and CFU-GM (P<0.001). CONCLUSION: The activated T lymphocyte may play an important role in the pathogenesis of SAA.

Adolescent↗

[Immunophenotyping study on the blood cells of the patients with paroxysmal nocturnal hemoglobinuria].

OBJECTIVE: To detect the expression of glycosyl-phosphatidyl-inositol (GPI) anchored protein on the blood cell membrane and its implication in the diagnosis of paroxysmal nocturnal hemoglobinuria (PNH). METHODS: The GPI anchored protein was detected by indirect immunofluorescence method with a panel of monoclonal antibodies (CD55, CD59, CD16, CD58, HIR2, CD15, CD2) in 22 PNH patients. RESULTS: The abnormal granulocytes and erythrocytes with deficient GPI anchored proteins were identified in all PNH patients whether the routine hemolysis tests for PNH, including Ham test and mCLST, were positive or negative. However, lymphocytes with PNH-defect were found in only 14 patients. CONCLUSION: GPI anchored protein assay provided sensitive and specific method for the diagnosis of PNH.

Adult↗

[Study on the immunophenotype of myeloid cells in myelodysplastic syndromes and its clinical implications].

OBJECTIVE: To explore the immunophenotype of myeloid cells in myelodysplastic syndromes and its clinical implications. METHODS: A panel of monoclonal antibodies was used to detect CD13, CD33, CD15 and CD14 on the membrane surfaces of myeloid cells in the bone marrow from 51 patients with myelodysplastic syndromes (MDS), 21 with aplastic anemia (AA), 21 with paroxysmal nocturnal hemoglobinuria (PNH), and 15 normal subjects, by immunoenzymatic assay. The morphology and chromosome karyotype of bone marrow cells of MDS patients were also examined. RESULTS: CD14+ cells, CD13+ cells and CD33+ cells in the bone marrow were more in MDS patients than in normal controls, AA patients and PNH patients. CD15+ cells in the bone marrow were less in MDS patients than in normal controls. The percentages of CD14, CD13 and CD33 positive cells in the bone marrow of MDS patients were related to the percentage of myeloblast, the chromosomal aberrations and the response to treatment. CONCLUSION: There was an immunophenotypic misexpression of myeloid cells in MDS patients. Immunophenotype analysis of myeloid cells might be useful for the diagnosis and directing treatment in MDS patients.

Adolescent↗

[Colony formation of bone marrow hematopoietic progenitor cells in patients with severe aplastic anemia and its relation to the results of immunosuppressive therapy].

OBJECTIVE: To explore the defect of hematopoietic stem cell and analyze the relationship between the colony formation capacity of bone marrow hematopoietic progenitor cells and the results of immunosuppressive therapy (IST) in severe aplastic anemia (SAA) patients. METHODS: Methylcellulose semisolid culture was used. RESULTS: Thirty patients with SAA at diagnosis were studied. In 90% of the patients, the CFU-E and CFU-GM yields were strikingly decreased and in 56.7% of the patients there was no colony formation at all. Nevertheless, there was still 10% of the patients having normal CFU-E, BFU-E or/ and CFU-GM yields. After IST, 59.1% of the patients showed colony formation improvement. The difference between the results obtained pre- and post-IST(for CFU-E and BFU-E, P < 0.01, for CFU-GM, P < 0.05) was significant. In the majority of the patients, the colony formation capacities were still under the normal post-IST. Moreover, 22.7% of the patients remained no colony formation post-IST. The IST response rates between the patients with or without colony formation had no statistically difference (80% versus 50%, P < 0.05). The increment of colony formation appeared later than the therapeutic effect did, but frequently concurred with the appearance of magkaryocytes in the bone marrow smear. CONCLUSION: In the majority of SAA patients, the hematopoietic stem cells or progenitors were defective perhaps caused by immune damage; IST can improve the hematopoiesis in vitro and in vivo of SAA patients.

Adolescent↗

[Determination of berberine in human serum by reversed-phase high performance liquid chromatography].

Berberine is a safe and effective anti-arrhythmic drug. We do not clearly know about the dose-effect relationship. Besides, berberine is not fit for administration by i.m. and i.v.. Therefore, it is necessary to build a PD model of berberine, study its pharmacodynamics parameters in vivo, and provide a concrete criteria in the clinical usage. In this article, we set up a method for determination of concentration of berberine in human serum by RP-HPLC with Lichrosorb RP-18 column (250 mm x 4 mm) and mobile phase of MeOH containing 0.15% triethylamine. The content of berberine was determined by the external standard method with UV detector at 347 nm. The linear range was 0.2-2 mg/L (r = 0.9996). The detectable limit was 2.55 ng. The average recovery was 90.73% and the coefficients of variation were below 8.0%. The method is sensitive and accurate.

Berberine↗

Different balance of excitation and inhibition in forward and feedback circuits of rat visual cortex.

Different cortical areas are linked reciprocally via forward and feedback connections. Forward connections are involved in the representation of retinal images, whereas feedback pathways may play a role in the selection and interpretation of visual information. To examine the synaptic mechanisms of forward and feedback connections between primary and secondary visual cortical areas directly, we have performed intracellular recordings in slices of rat visual cortex. Irrespective of stimulus intensity and membrane potential, 78% (45/58) of the cells in striate cortex activated by feedback input showed monosynaptic responses that were depolarizing only, and inhibitory inputs were evident merely as a slight acceleration in the decay of EPSPs. In contrast, in 89% (17/19) of the cells, stimulation of forward input evoked monosynaptic excitatory postsynaptic potentials (EPSPs), followed by disynaptic, hyperpolarizing inhibitory postsynaptic potentials (IPSPs). EPSPs followed by IPSPs also were recorded after stimulation of local connections within primary visual cortex (92%, 12/13) and after activation of thalamocortical input (91%, 10/11). These results suggest that the synaptic organization of feedback connections are distinct from forward, local, and thalamocortical circuits. The findings further indicate that intracortical back projections exert modulatory influences via synaptic mechanisms in which weak inhibitory input is strongly dominated by excitation.

Animals↗

Gramicidin A aggregation in supported gel state phosphatidylcholine bilayers.

Using an atomic force microscope, supported bilayers of saturated phosphatidylcholine (in the gel state) containing various amounts of gramicidin A (gA) were imaged in aqueous solutions and at room temperature. gA clusters were directly observed for the first time under these conditions. It was found that, at a lower gA concentration, gA aggregated into domains, composed of small clusters along with a considerable amount of lipids. This basic aggregation unit, most likely a hexamer, remained the same for acyl chain lengths from 14 to 18 carbons. These small clusters were observed to form elongated aggregates (line type) but never into extended pure gA domains. When gA concentrations were increased, for bilayers with 16 carbons or less, gA aggregated into larger domains but the basic unit remained separated by lipid molecules. At about 5 mol % gA, a percolation-like transition occurred at which the line type aggregates were connected to each other. However, for bilayers with more than 16 carbons, multiple lamellar structures were formed at higher gA fractions and the top layer had a ripple-like surface morphology. The molecular mechanism for the formation of these peculiar structures remains to be elucidated.

1,2-Dipalmitoylphosphatidylcholine↗

High resolution surface structure of E. coli GroES oligomer by atomic force microscopy.

Using atomic force microscopy (AFM) in aqueous solution, we show that the surface structure of the oligomeric GroES can be obtained up to 10 angstroms resolution. The seven subunits of the heptamer were well resolved without image averaging. The overall dimension of the GroES heptamer was 8.4 +/- 0.4 nm in diameter and 3.0 +/- 0.3 nm high. However, the AFM images further suggest that there is a central protrusion of 0.8 +/- 0.2 nm high and 4.5 +/- 0.4 nm in diameter on one side of GroES which displays a profound seven-fold symmetry. It was found that GroEL could not bind to the adsorbed GroES in the presence of AMP-PNP and Mg2+, suggesting that the side of GroES with the central protrusion faces away from the GroEL lumen, because only one side of GroES was observed under these conditions. Based on the results from both electron and atomic force microscopy, a surface model for the GroES is proposed.

Bacterial Proteins↗

Imaging biological structures with the cryo atomic force microscope.

It has long been recognized that one of the major limitations in biological atomic force microscopy (AFM) is the softness of most biological samples, which are easily deformed or damaged by the AFM tip, because of the high pressure in the contact area, especially from the very sharp tips required for high resolution. Another is the molecular motion present at room temperature due to thermal fluctuation. Using an AFM operated in liquid nitrogen vapor (cryo-AFM), we demonstrate that cryo-AFM can be applied to a large variety of biological samples, from immunoglobulins to DNA to cell surfaces. The resolution achieved with cryo-AFM is much improved when compared with AFM at room temperature with similar specimens, and is comparable to that of cryo-electron microscopy on randomly oriented macromolecules. We will also discuss the technical problems that remain to be solved for achieving even higher resolution with cryo-AFM and other possible applications of this novel technique.

Animals↗

Chaperonins GroEL and GroES: views from atomic force microscopy.

The Escherichia coli chaperonins, GroEL and GroES, as well as their complexes in the presence of a nonhydrolyzable nucleotide AMP-PNP, have been imaged with the atomic force microscope (AFM). We demonstrate that both GroEL and GroES that have been adsorbed to a mica surface can be resolved directly by the AFM in aqueous solution at room temperature. However, with glutaraldehyde fixation of already adsorbed molecules, the resolution of both GroEL and GroES was further improved, as all seven subunits were well resolved without any image processing. We also found that chemical fixation was necessary for the contact mode AFM to image GroEL/ES complexes, and in the AFM images. GroEL with GroES bound can be clearly distinguished from those without. The GroEL/ES complex was about 5 nm higher than GroEL alone, indicating a 2 nm upward movement of the apical domains of GroEL. Using a slightly larger probe force, unfixed GroEL could be dissected: the upper heptamer was removed to expose the contact surface of the two heptamers. These results clearly demonstrate the usefulness of cross-linking agents for the determination of molecular structures with the AFM. They also pave the way for using the AFM to study the structural basis for the function of GroE system and other molecular chaperones.

Adenylyl Imidodiphosphate↗

Biological cryo atomic force microscopy: a brief review.

Despite many successes, atomic force microscopy (AFM) of biological specimens at room temperature is still severely limited by at least two factors: the softness and the thermal motion of flexible multi-domain/subunit molecules. Both problems can be overcome by imaging biological structures at cryogenic temperatures. Even though the instrumentation is considerably more complex and earlier attempts were largely unsuccessful, cryo-AFM has recently been demonstrated on a number of biological specimens, using an AFM operated in liquid nitrogen vapor under ambient pressure. In this brief review, both the method of instrumentation and the latest biological applications are discussed. Not only has the cryo-AFM attained high resolution on those specimens that could not be well imaged at room temperature, but it has also produced potentially important information on several specimens. These results firmly establish the cryo-AFM as a useful and versatile structural probe in biology with its own unique capabilities.

Animals↗

Engineering new functions and altering existing functions.

Structural and mechanistic information, sequence comparisons, and site-directed mutagenesis data continue to provide a basis for the rational design of new protein functions and the alteration of existing functions. Random mutagenesis and 'directed evolution' approaches, however, are making significant headway in solving protein engineering problems, proving highly practical for tuning properties such as enzyme substrate specificity.

Binding Sites↗

The effect of deformation on the lateral resolution of atomic force microscopy.

A computer model based on the elastic properties of rubber is introduced for the evaluation of the lateral resolution in atomic force microscopy of deformable specimens. The computational results show that, if the full width at half-height can be defined as the lateral resolution, it is continuously improved at greater probe forces, at the expense of a reduced molecular height, In fact, even for a probe that is bigger than the molecule, the real size of the molecule can be 'recovered' at about 25% compression. This result demonstrates that for a better lateral resolution, a greater probe force can be beneficial, provided that the molecule is not moved or damaged and the response remains elastic. Measurements on isolated low-density lipoproteins (LDL) show that with 26% vertical compression, the lateral size measured in atomic force microscopy is only 72% of the value predicted by a simple convolution, and is only slightly larger (approximately 13%) than the known size of LDL. Therefore, the results on LDL provide a direct support for the conclusions of the computational model.

Computer Simulation↗

Reperfusion injury on cardiac myocytes after simulated ischemia.

The extent of cardiac injury incurred during reperfusion as opposed to that occurring during ischemia is unclear. This study tested the hypothesis that simulated ischemia followed by simulated reperfusion causes significant "reperfusion injury" in isolated chick cardiomyocytes. Cells were exposed to hypoxia, hypercarbic acidosis, hyperkalemia, and substrate deprivation for 1 h followed by 3 h of reperfusion. Irreversible cell membrane injury, measured by propidium iodide uptake, increased from 4% of cells at the end of ischemia to 73% after reperfusion; death occurred in only 17% of cells kept ischemic for 4 h. Lactate dehydrogenase release was consistent with these changes. Lengthening ischemia from 30 to 90 min increased cell injury as expected, but of the total cell death, > 90% occurred during reperfusion. "Chemical hypoxia" composed of cyanide (2.5 mM) plus 2-deoxyglucose augmented injury before reperfusion compared with simulated ischemia. Inhibition of oxygen radical generation by use of metal chelator 1,10-phenanthroline reduced cell death from 73% to 40% after reperfusion (P = 0.001). We conclude that simulated reperfusion significantly augments the cellular membrane damage elicited by simulated ischemia in isolated cardiomyocytes devoid of other factors and suggest that reactive oxygen species, perhaps from the mitochondria, participate in this injury.

Animals↗

[Retinoic acid nuclear receptor beta (RAR beta) inhibits breast carcinoma growth].

Retinoids are capable of modulating cellular differentiation and proliferation. Retinoids mediate gene function through a series of nuclear receptors. The retinoic acid receptor beta (RAR beta) has been shown to play an important role in the differentiation of a number of cell types. RAR beta is either absent or expressed at extremely low levels in a number of tumor types including breast carcinoma. It was demonstrated that transfection of RAR beta gene in breast carcinoma cell with its subsequent expression resulted in inhibition of cell growth. Retinoic acid significantly inhibited monolayer growth of the breast carcinoma cells expressing RAR beta, while it had no effect on the growth of the control cells. The RAR beta expressing cells formed much smaller and fewer colonies in soft agar and were significantly less tumorigenic in nude mice than the controls. These results suggest that RAR beta may function as a tumor suppressor in breast carcinoma cells.

Animals↗

[Growth inhibitory effect of retinoic acid in human breast cancer cells correlates with retinoic acid receptor alpha (RAR alpha) expression].

The relationship between the effect of retinoic acid (RA) on the growth of breast cancer cell and their estrogen receptor (ER) status as well as the relationship between RA effect and the expression of retinoic acid receptorsd (RAR alpha) were studied by cell growth assay, Northern Blot and gene transfection. It was found that RA could only inhibit the growth of ER-positive but not ER-negative breast cancer cells. RAR alpha mRNA level was significantly higher in ER-positive breast cancer cell lines than that in ER-negative breast cancer cell line. The expressions of other subtypes of RAR in ER-positive cells were not significantly different from those in ER-negative ones. When, RAR alpha cDNA was introduced and ixpressed in RA-resistant, ER-negative MDA-MB-231 breast cancer cell line, its growth was strongly inhibited by RA. These results indicate that RAR alpha plays a major role in the retinoi-dmediated inhibition of growth in human breast cancer cells.

Antineoplastic Agents↗

Inhibition of spontaneous apoptosis in human breast cancer.

Breast tumorigenesis proceeds through an accumulation of specific genetic alteration. Breast malignant transformation is dependent on not only the rate of cell production but also on apoptosis, a genetically programed process of autonomous cell death. We investigated whether breast tumorigenesis involved an altered susceptibility to apoptosis and proliferation by examining normal breast epithelium and breast cancer samples. We found there is a great inhibition of spontaneous apoptosis in breast cancer cells compared with normal breast epithelium. The inhibition of apoptosis in breast cancer may contribute to neoplastic transformation.

Apoptosis↗