Biomedical subjects
Publications and source records attributed to Q Guo.
In the film sample analysis, the consstence within standard sample and analytical sample is discussed by emission spectral method. Using spark light source and moving electrode, the content of Sn/In in the ITO film is obtained by repeatedly exposing four samples. The accuracy and precision are satisfying.
OBJECTIVE: To make a hereditary analysis of varicose vein of lower extremities(VVLE), including hereditary pattern, sex ratio and the transmitted expression male and female of varicose vein of lower extremities. METHODS: The ascertainment probability was estimated according to Weinberg's proband method. The value of segregation ratio P and proportion of sporadic cases X were estimated by the maximum likelihood score method. The calculations were performed using computer program of segregation analysis. The VVLE cases were random samples from inpatients and outpatients of the authors' hospital from 1991 to 1996. RESULTS: There were 68 pedigrees including 80 nuclear families in 3 different mating types:UAU, UAA and AAA. UAU, UAA and AAA were analysed together. UAA and UAU (multiplex families) were analysed respectively. All the values were compatible with autosomal dominant inheritance of incomplete penetrance; the penetrance frequencies were 91.94%, 78.14% and 69. 92% respectively. The proportion of X was 24.32%. The UAU was compatible with autosomal recessive inheritance; the proportion of sporadic cases was 36.75%. CONCLUSION: VVLE is compatible with autosomal dominant and recessive inheritance. The sex ratio of male:female is 2.39:1. The transmission is possible from male and female to male and female both.
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Mutations in presenilin-1 (PS-1) account for approximately half the cases of autosomal dominant early-onset Alzheimer's disease (AD). Recent data indicate that PS-1 mutations may render neurons vulnerable to apoptosis induced by various insults. We now report that 17beta-estradiol, which appears to reduce the risk of sporadic AD, protects cultured PC12 cells expressing mutant PS-1 against apoptosis induced by trophic factor withdrawal (TFW) and exposure to amyloid beta-peptide 25-35 (Abeta). Estriol also provided significant protection against apoptosis induced by TFW and Abeta, whereas corticosterone was ineffective. 17beta-Estradiol prevented decreases in mitochondrial transmembrane potential and energy charge/redox state following exposure of cells to TFW and Abeta in control cell lines and lines expressing mutant PS-1, suggesting an action in the apoptotic pathway upstream of mitochondrial alterations. Abeta caused an increase in mitochondrial reactive oxygen species which was enhanced by mutant PS-1, and suppressed by 17beta-estradiol. The ability of 17beta-estradiol to preserve mitochondrial function, suppress oxidative stress, and counteract the pro-apoptotic actions of mutant PS-1 suggests a generalized neuroprotective action of estrogens in both sporadic and inherited forms of AD.
Mutations in genes encoding presenilin-1 (PS-1) and presenilin-2 (PS-2) cause many cases of autosomal dominant inherited forms of early-onset Alzheimer's disease (AD). PSs are expressed in neurons throughout the nervous system, with differences in abundance among cell populations. PS-1 and PS-2 each have six to eight transmembrane domains and are localized mainly in the endoplasmic reticulum (ER). PSs may interact with cytoskeletal proteins and beta-amyloid precursor protein (APP) in ways consistent with roles in membrane trafficking and APP processing. Expression of mutant PSs in cultured cells and transgenic mice results in increased production of an amyloidogenic-cytotoxic form of amyloid beta-peptide (Abeta). Neural cells expressing mutant PSs exhibit increased sensitivity to apoptosis induced by trophic factor withdrawal and Abeta. The proapoptotic action of mutant PSs involves perturbed calcium release from ER stores and increased levels of oxidative stress. PS mutations may also suppress neurotransmitter synthesis in cholinergic neurons, suggesting a role in regulation of neuronal phenotype. Homology of PSs with the C. elegans gene sel-12 and phenotypic similarities of PS-1 and Notch knockout mice suggest a developmental role for PSs in somitogenesis. Collectively, the emerging data suggest intriguing roles of PSs in neuronal plasticity and cell death and highlight the importance of the ER as a regulatory site involved in the pathogenesis of neuronal degeneration in AD.
The human blood platelet fibrinogen receptor, integrin alphaIIbbeta3 (glycoprotein IIb-IIIa) is an archetypal member of the integrin family of adhesive molecules and is the only integrin encoded by genes physically linked in the genome. Because studies on the normal and abnormal expression of any gene require a thorough understanding of its organization, the initial goals of the current study were to determine the size and complete the genomic organization for the beta3 gene. We now report the isolation of the entire beta3 gene in a single P1 plasmid and for the first time have linked the first and second exons on a contiguous fragment of DNA. Using pulsed-field gel analysis, we determined the full size of the beta3 gene to be 63 kb and show a large (16.7 kb) first intron; based on this information, we propose a uniform numbering system for the beta3 exons. We have completed the 5' genomic structure and generated a long-range restriction map. The promoter and the 5' end of the first intron were found to have approximately 50% sequence identity with a region of the avian beta3 gene known to possess functional transcriptional activity. Analysis of three different homologous regions led to the identification of a sequence in the 5'-UTR of the human gene, CCGCGGGAGG, which shares 90% identity with the avian gene and which bound nuclear proteins in DNaseI and electrophoretic mobility shift assay studies. Mutating this sequence caused a 2.6-fold reduction in reporter gene activity. In these studies we have (1) determined the full length and 5' organization of the beta3 gene, (2) identified a large region of homology between the 5' regions of the avian and human genes, and (3) identified a sequence in the 5'-UTR that augments gene expression. Knowing the genomic structure of beta3 has permitted the uncovering of new mechanisms of mutagenesis causing Glanzmann thrombasthenia (Jin et al, J Clin Invest 98:1745, 1996), and our findings will be valuable for such genetic analyses as well as for studies on the transcriptional regulation of beta3 and other integrin genes.
The cerebellum is essential for fine motor control of movement and posture, and its dysfunction disrupts balance and impairs control of speech, limb and eye movements. The developing cerebellum consists mainly of three types of neuronal cells: granule cells in the external germinal layer, Purkinje cells, and neurons of the deep nuclei. The molecular mechanisms that underlie the specific determination and the differentiation of each of these neuronal subtypes are unknown. Math1, the mouse homologue of the Drosophila gene atonal, encodes a basic helix-loop-helix transcription factor that is specifically expressed in the precursors of the external germinal layer and their derivatives. Here we report that mice lacking Math1 fail to form granule cells and are born with a cerebellum that is devoid of an external germinal layer. To our knowledge, Math1 is the first gene to be shown to be required in vivo for the genesis of granule cells, and hence the predominant neuronal population in the cerebellum.
Mutations in the presenilin genes (PS-1 and PS-2) are linked to early onset familial Alzheimer's disease (AD), but the mechanisms by which these mutations cause the cognitive impairment characteristic of AD are unknown. Basal forebrain cholinergic neurons are involved in learning and memory processes, and reductions in choline acetyl-transferase (ChAT) activity are a characteristic feature of AD brain. We therefore hypothesized that presenilin mutations suppress expression of the cholinergic phenotype. In rat PC12 cells stably transfected with the human PS-1 gene containing the Leu --> Val mutation at codon 286 (L286V), we observed a drastic reduction (>90%) in basal ChAT activity compared with cells transfected with vector alone. By immunocytochemistry, a similar decrease in ChAT protein levels was found in the mutant transfectants. In cells differentiated with nerve growth factor, ChAT activity was again markedly lower in L286V-expressing cells than in control cells. We also observed reductions in ChAT activity in PC12 cells expressing the wild-type human PS-1 gene but to a lesser extent than in L286V-expressing cells. The viability of cells transfected with either the wild-type or the mutant PS-1 gene was not compromised. Our results suggest that PS-1 mutations may contribute to the cognitive impairment in AD by causing a nontoxic suppression of the cholinergic phenotype.
Prostaglandin H synthase (PGHS) catalyzes a key step in the biosynthesis of a variety of bioactive lipid mediators. The two known isoforms (PGHS-1 and -2) share about 60% amino acid identity, but exhibit distinct interactions with substrates, activators, and inhibitors. Ovine PGHS-1 has previously been shown to have a distinctive protease-sensitive site near Arg277; cleavage by trypsin, chymotrypsin, or proteinase K produces fragments of 33 and 38 kDa and loss of activity. The ovine PGHS-1 crystal structure shows Arg277 located in an exposed loop structure; homology modeling predicts similar loop structures for both human isoforms (hPGHS-1 and -2). We have used limited proteolytic digestion of recombinant hPGHS-1 and hPGHS-2 to probe their structures. Incubation of hPGHS-1 with either trypsin or proteinase K produced 33- and 38-kDa fragments and loss of activity. In contrast, incubation of hPGHS-2 with the same proteases led to cleavage of only a 2- to 3-kDa fragment, with no decrease in activity. Immunoblotting with site-specific antibodies demonstrated that the cleaved fragment originated from the hPGHS-2 C-terminus. Similar immunoblotting experiments indicated that trypsin did not attack the ovine PGHS-1 C-terminus. Mutagenesis was used to replace Pro263 of hPGHS-2 (corresponds to Arg277 of ovine PGHS-1) with arginine, inserting a potential trypsin site. Incubation of this P263R hPGHS-2 mutant with either trypsin or proteinase K resulted in cleavage near the C-terminus and retention of activity, just as with wild-type hPGHS-2. A peptide containing residues 259-268 of the P263R mutant was cleaved by trypsin at the same rate as a peptide corresponding to hPGHS-1 residues 272-281, demonstrating that the sequence differences were not responsible for the lack of tryptic cleavage at residue 263 in the hPGHS-2 mutant. Preincubation of hPGHS-2 with graded levels of guanidinium HCl before incubation with proteinase K did not produce large proteolytic fragments, indicating that the hPGHS-2 loop region was not selectively unfolding. The results point to two regions of significant structural difference between PGHS-1 and -2: the Arg277 loop, which is protease-sensitive in PGHS-1 but protease-resistant in PGHS-2, and the C-terminus, which is protease-sensitive in PGHS-2 but not in PGHS-1.
Most autosomal dominant inherited forms of early onset Alzheimer's disease (AD) are caused by mutations in the presenilin-1 (PS-1) gene on chromosome 14. PS-1 is an integral membrane protein with six to nine membrane-spanning domains and is expressed in neurons throughout the brain wherein it is localized mainly in endoplasmic reticulum (ER). The mechanism or mechanisms whereby PS-1 mutations promote neuron degeneration in AD are unknown. Recent findings suggest links among deposition of amyloid beta-peptide (Abeta), oxidative stress, disruption of ion homeostasis, and an apoptotic form of neuron death in AD. We now report that expression of the human PS-1 L286V mutation in PC12 cells increases their susceptibility to apoptosis induced by trophic factor withdrawal and Abeta. Increases in oxidative stress and intracellular calcium levels induced by the apoptotic stimuli were exacerbated greatly in cells expressing the PS-1 mutation, as compared with control cell lines and lines overexpressing wild-type PS-1. The antiapoptotic gene product Bcl-2 prevented apoptosis after NGF withdrawal from differentiated PC12 cells expressing mutant PS-1. Elevations of [Ca2+]i in response to thapsigargin, an inhibitor of the ER Ca2+-ATPase, were increased in cells expressing mutant PS-1, and this adverse effect was abolished in cells expressing Bcl-2. Antioxidants and blockers of calcium influx and release from ER protected cells against the adverse consequences of the PS-1 mutation. By perturbing cellular calcium regulation and promoting oxidative stress, PS-1 mutations may sensitize neurons to apoptotic death in AD.
Transgenic technology affords exciting new opportunities in the field of behavioral neuroendocrinology. We have extended our research into the behavioral function of oxytocin in maternal and social behavior using two transgenic approaches: (i) targeted deletion of the oxytocin gene in mice and (ii) augmented oxytocin receptor expression in the brain. Mice genetically deficient in oxytocin can mate, give birth, and display normal maternal behavior; however, milk ejection and certain aspects of social behavior are affected. Comparative studies of oxytocin receptors have led to the observation that species differences in social organization are associated with differences in receptor distribution. Specifically, monogamous prairie voles and nonmonogamous, asocial montane voles exhibit different patterns of OT receptor expression in the brain. Transgenic mice have been created with a reporter gene driven by the prairie vole oxytocin receptor gene promoter. Analysis of the expression pattern suggests that it should be possible to manipulate receptor expression in the vole brain in order to examine the effects of receptor distribution on behavior.
A simple, rapid method to isolate cardiac myocytes from rat and guinea-pig was described by using this method. Morphologically intact and calcium tolerant single myocytes were obtained. The myocytes were suited well for various electrophysiological recording especially in patch clamp study.
Monolayer culture of spontaneously contracting rat heart cells was obtained from newborn SD rats and seeded into culture plates. 100TCID50 of coxsackie virus B3(CVB3) (Nancy strain) was added as the infected group. The dynamics of taurine transport across membrane of normal heart cells and changes after infection were examined by using radioactive isotope tracing techniques. The results demonstrated that: (1) two uptake systems of taurine with different affinities and capacities were present in the plasma membrane of cultured rat cardiomyocytes, the Km of high and low affinity were 9.5 x 10(-5) mol/L and 1.3 x 10(-2) mol/L, respectively; (2) taurine transport across membrane was sodium-dependent with a positive linear relationship between amount of taurine influx and extracellular concentration of sodium; in addition, it was also regulated by osmolarity; (3) beta-alanine, a carboxylic analog of taurine, could inhibit the uptake of taurine in dose-dependently; (4) taurine influx was decreased in cultured rat cardiomyocytes after infected with CVB3. (5) the Inhibition ratio of taurine transport was enhanced as the infected time prolonged. This experiment provides a basis for further investigation of kinetic test of taurine transport and more study should be carried out to learn the effect of taurine in viral myocarditis.
Enhancement effect of easily volatile elements I (boiling point = 184.35 degrees C) and S (boiling point = 444.67 degrees C) on spectral line intensities of various trace elements in laser source plasma were investigated by experiments. In order to discuss mechanism of enhancement effect, some of the plasma parameters (Texc,ne) were measured.
Quantitative determination of the nickel, tin, copper, scandium, zirconium and hafnium in silicate sample were carried out by laser microspectrochemical analysis, results show that the method has good precision and accuracy. The stability of the laser output and the selection of auxiliary spark excitation condition in experimental process were discussed.
To ascertain whether complete cerebral ischemia-reperfusion activate L-Arg: NO pathway in canine brain, we anestherized nine adult dogs with ketamine and fentayle and randomly divided into two groups. Four dogs were nonischemic control group. Five dogs were complete cerebral ischemia-reperfusion group, they underwent a 18-minute cardiac arrest, and were resusciatation by standard CPR, supported by intensive care for 8 hours. At the end of each experiment, the parietal cortex was assayed for content of Nitrite and NADPH-positive neurons. Compared with the control group, the contents of Nitrite and NADPH-positive neurons of coxtex in complete cerebral ischemia-reperfusion group increased significantly (P < 0.01). The results suggest that complete cerebral ischemia-reperfusion activate the L-Arg: NO pathway in canine brain, and NO may play an important role in cerebral ischemia-reperfusion injury.
Mutations in the presenilin-1 (PS-1) gene on chromosome 14 are linked to autosomal dominant early-onset Alzheimer's disease. The amino acid sequence of PS-1 predicts an integral membrane protein and immunocytochemical studies indicate that PS-1 is localized to endoplasmic reticulum (ER). We report that expression of PS-1 mutation L286V in cultured PC12 cells exaggerates Ca2+ responses to agonists (carbachol and bradykinin) that induce Ca2+ release from ER. Cells expressing L286V exhibit enhanced elevations of [Ca2+]i following exposure to amyloid beta-peptide (A beta) and increased vulnerability to A beta toxicity. An antagonist of voltage-dependent calcium channels (nifedipine), and a blocker of Ca2+ release from ER (dantrolene), counteract the adverse consequences of the PS-1 mutation. By perturbing Ca2+ homeostasis, PS-1 mutations may sensitize neurons to A beta-induced apoptosis.
The comparison of the protective effects of four components of "green tea polyphenols' (GTP) - (-)-epigallocatechin gallate, EGCG; (-)-epicatechin gallate, ECG; (-)epigallocatechin, EGC; and (-)epicatechin, EC - against iron-induced lipid peroxidation in synaptosomes showed that: (1) the inhibitory effects of those compounds on TBA reactive materials from lipid peroxidation decreased in the order of EGCG > ECG > EGC > EC; (2) the scavenging effects of those compounds on lipid free radicals produced by lipid peroxidation could be classified as follows: ECG > EGCG > EC > EGC. Furthermore, we investigated the iron-chelating activity and the free radical scavenging activity of those compounds as their protective mechanisms against lipid peroxidation in synaptosomes. As for the iron-chelating activity, the ratio of EGC, EGCG, ECG or EC to iron(III) was 3:2, 2:1, 2:1 and 3:1, respectively. The hydroxyl radical (HO) scavenging activity of those compounds was investigated in a photolysis of the H2O2 system. It was found that their ability to scavenge hydroxyl radicals decreased in the order of ECG > EC > EGCG >> EGC. It was also found that they could scavenge lipid free radicals in the lecithin/lipoxidase system and their scavenging activity was classified as follows: ECG > EGCG >> EGC > EC. Moreover, we found that their antioxidant active positions were different from each other and the stability of the semiquinone free radicals produced by those compounds in NaOH solution decreased in the order of EGCG > ECG >> EC. The results indicated that the ability of those compounds to protect synaptosomes from the damage of lipid peroxidation initiated by Fe2+/Fe3+ was dependent not only on their iron-chelating activity and free-radical scavenging activity, but also on the stability of their semiquinone free radicals.