Biomedical subjects
Publications and source records attributed to B Zhao.
Cultured endothelial cells from bovine aorta were exposed to oxidized low density lipoprotein and examined by electron microscopy. The endothelial cells contracted slightly and the intercellular junctions became unclear. Some osmiophilic material increased in the cytoplasm. The oxidized low density lipoprotein appears to injure endothelial cells and thereby plays a causative role in atherogenesis and thrombogenesis.
High-density lipoprotein (HDL) of 100-400 micrograms/ml did not prevent morphological alterations of human blood platelets treated with serotonin (1-5 microM). Highly concentrated HDL (1,200 micrograms/ml) appeared to activate platelets in vitro. These findings indicate that whole HDL may not inhibit agonist-induced platelet activation.
Ornithine decarboxylase (ODC), the key regulatory enzyme in mammalian polyamine biosynthesis, is rapidly induced by mitogens and tumor promoters. We used transient expression assays and DNA-protein binding studies to examine the regulation of ODC promoter activity by phorbol esters and serum growth factors. A fragment of the ODC 5' flanking region (nt-1156 to +13) was sufficient to confer 12-O-tetradecanoylphorbol-13-acetate (TPA)-responsive expression to a luciferase reporter gene when transfected into H35 cells. However, induction by TPA was not observed in Rat2 fibroblasts, although refeeding of serum-starved Rat2 cells with fresh serum-containing medium rapidly induced a fivefold to sixfold increase in ODC promoter activity, maximal about 8 h after refeeding. Deletion analysis demonstrated that several sequences contributed to basal ODC promoter activity but that nt -92 to +13 was sufficient for induction by TPA or by serum. This sequence lacked canonical TPA-responsive elements, and an activator protein-1 (AP-1) consensus oligonucleotide failed to compete effectively for proteins binding to this region. Two of four protein complexes observed by gel-shift analysis of nt -92 to +13 were competitively inhibited by wild-type but not mutant oligonucleotides encompassing a variant cyclic AMP-response element (CRE) (ODC nt -50 to -42); however, a consensus CRE did not compete. Mutagenesis of this site demonstrated that it contributes to basal expression of the ODC promoter but not to TPA or serum responsiveness. Thus, we conclude that the proximal ODC promoter (nt -92 to +13) responds to TPA and serum stimulation in a cell-type-specific manner that is not mediated by canonical AP-1 elements.
The D-mef2 gene encodes a MADS domain transcription factor expressed in differentiated muscles and their precursors in the Drosophila embryo. Embryos deficient for D-MEF2 protein due to a deletion of upstream transcriptional control sequences fail to form muscle, suggesting that the gene is required for muscle cell differentiation. To directly demonstrate a role for D-mef2 in embryonic myogenesis, we isolated gene mutants containing EMS-induced point mutations, characterized the effects of these mutations on D-MEF2 protein stability and nuclear localization, and analyzed the resulting muscle phenotypes. Our results show that in the somatic muscle lineage, D-mef2 is required for both the formation and patterning of body wall muscle. In the absence of somatic myogenesis, there is extensive apoptosis among the myoblast cell population. In contrast, in the cardiac muscle lineage, morphogenesis of the dorsal vessel occurs normally but the three myosin subunit genes are not expressed. Mutant embryos also exhibit an abnormal midgut morphology, which correlates with the absence of alpha PS2 integrin gene expression and muscle-specific enhancer function, suggesting that D-mef2 regulates the inflated locus which encodes this integrin subunit. D-MEF2 is also expressed in adepithelial cells and rare D-mef2 transheterozygous mutant adults fail to fly, consistent with defects observed in the indirect flight muscles. These results demonstrate that the D-mef2 gene has multiple functions in myogenesis and tissue morphogenesis during Drosophila development.
The adhesion of human blood platelets is studied with an in vitro model using reflection contrast microscopy and an image analysis system. The adhesive feature is promoted by oxidatively modified low density lipoprotein, which also induces functional morphological changes of platelets. However, when washed platelets are pretreated with 0.05 mM cytochalasin D, oxidized low density lipoprotein (100 micrograms/ml) causes a slower increase of the adhesion area (11.6 microns 2/min) compared to untreated platelets (15.7 microns 2/min) or platelets treated by oxidized low density lipoprotein alone (20.5 microns 2/min, P < 0.01). These results are supported by light transmission analysis and by transmission electron microscopy. Our experiments suggest that cytochalasin D inhibits the change of platelets in shape induced by oxidized low density lipoprotein, hinders the adhesion, but does not prevent the adhesion entirely.
This paper introduces a program written on the image analysis system VIDAS 2.5. It enables the automatic quantification of high numbers of adhesion areas of vital human platelets, thus allowing statistical analysis. These adhesion areas were observed by reflection contrast microscopy (RCM), which generates images of an intense contrast and serves as a prerequisite for an evaluation by image analysis. However, RCM-photographs of the observed platelets have highly varying mean greyvalues and greyranges. These common problems for self-operating identification are excluded by two procedures within the program: 1. calibration of the scanning process for an optimal use of the available greyvalues provided by the negative, camera, and the image analysis system; and 2. relation of the threshold for discrimination of adhesion areas to the statistic parameters of the histogram within each individual digitized image. Images processed according to these prerequisites were transferred to the VIDAS implemented routines for identification and measurement of areas. Thus, image analysis combined with RCM offers a tool for basic and clinical platelet research, which is shown by an example of stimulation and inhibited stimulation of platelet activation.
In a Caucasian woman, a single vein was found which originated from the inferior vena cava and passed behind the abdominal aorta to join the left ovarian vein just before its entry into the left renal vein. An extra connection therefore existed between the left kidney and the inferior vena cava.
We describe a structural validation of the use of presentation scaffolds for control and elucidation of bioactive conformations of peptides. The protein REI-RGD34--produced by inserting the sequence RIPRGDMP into the CDR1 loop region of the immunoglobulin VL domain REI--strongly inhibits fibrinogen binding to the integrins alpha IIb beta 3 and alpha V beta 3. In the X-ray crystal structure of their protein at 2.4 A resolution, the RGD-containing loop exhibits defined electron density that is consistent with models for the bioactive conformations of ligands of these receptors based on previous small-molecule studies. Furthermore, a search of a small-molecule database with conformational information derived from the structure of REI-RGD34 identified constrained peptides and peptidomimetics known to be antagonists of the platelet receptor alpha IIb beta 3.
Several xenobiotic metabolizing enzymes, including CYP1A1, NAT2 and GSTM1, are subject to genetic polymorphisms. Because these enzymes are important for the detoxification and/or bioactivation of drugs and carcinogens, these polymorphisms have important implications in therapeutics and cancer susceptibility. The distributions of CYP1A1, NAT2 and GSTM1 genotype frequencies in unrelated individuals of the Indian (n = 139) and Malay (n = 146) populations were characterized by the polymerase chain reaction. The respective allelic frequencies of wild-type and mutant alleles of CYP1A1 were 0.82 and 0.18 for the Indians, and 0.69 and 0.31 for the Malays. The frequencies of wild-type, M1, M2 and M3 of NAT2 among Indians were 0.44, 0.20, 0.32 and 0.04 respectively. The corresponding NAT2 allelic frequencies in Malays were 0.41, 0.12, 0.38 and 0.09. The GSTM1*A allele could not be amplified in 33.1% of Indians and 61.6% of Malays. At least one GSTM1*B allele was detected in 7.2% and 7.5% of the respective populations. The allelic frequencies of CYP1A1, NAT2 and GSTM1 among Malays are similar to previously reported frequencies among Chinese in the region. These findings will be of importance in the determination of cancer risks in these populations.
Using electron spin resonance (ESR) spin trapping technique, we found that baicalin (B) could scavenge hydroxyl radicals generated from Fenton reaction. It also could scavenge superoxide radicals generated from the reaction system containing xanthine (X) and xanthine oxidase (XO), as was found by using chemiluminescence (CL) method. Kinetic studies on the competition between baicalin and a spin trap 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) in trapping OH showed that baicalin had a kinetic reactive rate constant of the order of 7.7 x 10(11) M-1 s-1 in its reaction with OH, and the studies on the competition between baicalin and ferricytochrome c (f.c.) in trapping O2- gave a kinetic reactive rate constant of 3.2 x 10(6) M-1 s-1 for baicalin in its reaction with O2-. Furthermore, we have investigated the protective effects of baicalin on erythrocyte membranes from hydroxyl free radical injuries. The results showed that baicalin could reduce hydrogen peroxide-induced hemolysis, protect the conformation of sulfhydryl groups (-SH) on membrane proteins and the membrane fluidity of erythrocytes incubated with hydrogen peroxide. The results indicated that baicalin could protect the membranes of erythrocytes from free radical injuries, and it was even more effective than alpha-tocopherol.
Through the screening on scoliosis among 24,130 school children in Hu Guan County, Shanxi Province, it showed that the forward-bending test was positive in 1,794 children (7.4 per cent), with male 3.64 per cent and female 3.8 per cent. The hump inclination angles that were more than 4 degrees were found in 665 children (2.75 per cent). The Cobb angles that were more than 10 degrees were found in 347 children. The incidence rate was 1.44 per cent, with a male to female ratio of 1:1.17. Most of them were idiopathic scoliosis.
Crown galls were induced by direct infection of sterile seedlings with Agrobacterium tumefaciens C58, and the transformation was proved by opine identification. The crown galls grew well in a hormone-free medium. B5 and MS basic media were good for the growth of crown galls (strain Ca), which increased up to 102 and 90 times in a month, respectively. However, 67-V and WP basic media are good for tanshinone production. It has been shown that crown galls can be utilized as a culture system to produce secondary metabolites.
The contents of panaxadiol and panaxatriol in Shihu Yeguang Pills manufactured by pharmaceutical works in Beijing, Jinan, Guangzhou and other places were determined by TLC-densitometry. A quantitative method has been developed to serve as a reference to quality control of these pills.
OBJECTIVE: For morphologic and functional investigations on the physiology and pathology of human blood platelets, their adhesion plays an important role. This feature is normally assessed concerning qualitative aspects, but there is a lack of quantitative investigations. STUDY DESIGN: We developed a model to quantify in vitro manipulations on thrombocytes by measuring their adhesion areas automatically. This was achieved by combining the VIDAS 2.5 image analysis system with reflection contrast microscopy (RCM). The evaluation was performed independent of the position of the grey value histogram by analyzing each digitized image corresponding to its individual mean grey value. RESULTS: The applicability and validity were verified from the example of a pharmacologic experiment administering the platelet stimulant oxidized low density lipoprotein (ox-LDL) to thrombocyte suspensions and confirming it by aggregometry. We assessed the temporal increase in adhesion area and mean total adhesion area of activated platelets, comparing unstimulated to stimulated ones. Ox-LDL accelerated the adhesion process and increased the adhesion area. CONCLUSION: The combination of RCM with consecutive image analysis provides useful parameters for in vitro examination on vital human platelets. Large amounts of data can be accumulated in a short time for an evaluation of influences of and on blood platelets.
The purpose of this study was to verify the role of somatostatin (Som) of brain in analgesia of electroacupuncture (EA). Intracerebroventricular (icv) injection of Som caused a more marked elevation of pain threshold and of EA analgesic effect, but the activity of Ca(2+)-APTase in hippocampus was significantly decreased; The Som and GABA levels in hippocampus and brain stem were decreased by EA analgesia. The Som in hippocampus and brain stem were obviously depleted by icv injection of cycteamine, but without any change of pain threshold and analgesic effect of EA. These results indicated that the exogenous Som of brain potentiated the analgesic effect of EA, however, the decrease of endogenous of some brain regions took part in the process of EA analgesia.
The rational design and synthesis of a highly potent inhibitor of HIV-1 protease have been accomplished. The inhibitor, SB 206343, is based on a model derived from the structure of the MVT-101/HIV-1 protease complex and contains a 4(5)-acylimidazole ring as an isosteric replacement for the P1'--P2' amide bond. It is a competitive inhibitor with an apparent inhibition constant of 0.6 nM at pH 6.0. The three-dimensional structure of SB 206343 bound in the active site of HIV-1 protease has been determined at 2.3 A resolution by X-ray diffraction techniques and refined to a crystallographic discrepancy factor, R (= sigma parallel Fo magnitude of/Fc parallel/sigma magnitude of), of 0.194. The inhibitor is held in the enzyme by a set of hydrophobic and polar interactions. N-3 of the imidazole ring participates in a novel hydrogen-bonding interaction with the bound water molecule, demonstrating the effectiveness of the imidazole ring as an isosteric replacement for the P1'--P2' amide bond in hydroxyethylene-based HIV-1 protease inhibitors. Also present are hydrogen-bonding interactions between N-1 of the imidazole ring and the carbonyl of Gly-127 as well as between the imidazole acyl carbonyl oxygen and the amide nitrogen of Asp-129, exemplifying the peptidomimetic nature of the 4(5)-acylimidazole isostere. All of these interactions are in qualitative agreement with those predicted by the model.
The effect of oxidized low density lipoprotein (LDL) on the functional morphology of human platelets in vitro was studied by means of transmission electron microscopy. The washed platelets, stimulated by oxidized LDL (50-300 micrograms protein/ml), showed disc-sphere transformation, centralization of granules and complete degranulation in a dose- and time-dependent manner. A cytodamage in platelet membrane was induced by oxidized LDL leading to a lower electron density of cytoplasm compared to control. The morphological observations were supported by an analysis of the platelet shape-change parameter. Since the shape change, induced by oxidized LDL (50 micrograms/ml), was inhibited by a preincubation of platelets with staurosporine (10 nM), the protein kinase C was probably involved in the platelet activation initiated by oxidized LDL. The present results suggest that oxidized LDL could contribute to pathological thrombosis and atherogenesis by activating platelets.
We report the first demonstration of EPR spectroscopy to study free radical reactions in live mice and excised muscle tissue resulting from the metabolism of nitrosobenzene. A broad three-line EPR spectrum (aN = 11.6 G) appeared in the buttocks region of a mouse place in an L-band loop gap resonator after intramuscular or intraperitoneal injection of 0.2 mmol/kg nitrosobenzene. The signal intensity reached a maximum at 20 to 30 min and remained constant well beyond 2 h. If muscle tissue was dosed with nitrosobenzene and excised within 5 min, a similar three-line X-band EPR spectrum was obtained which was preceded by the rapid growth and subsequent decay of an EPR spectrum identical with that of the phenylhydronitroxide radical, which was presumably generated from reactions between nitrosobenzene and reducing agents in the blood or tissue such as NADH or ascorbic acid. A model system containing nitrosobenzene and unsaturated fatty acids (olive oil or animal fat) yielded an identical three-line spectrum resulting from radical adducts of nitrosobenzene across the double bond. Overall, these results suggest that the most probable mechanism in vivo was nitrosobenzene covalently adding ("binding") to polyunsaturated fatty acid clusters in fat or membranes.