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Biomedical subjects

L Wu

Publications and source records attributed to L Wu.

At least 523 records · Page 29Linked to original sources
PubMed1996

Selective defects in the development of the fetal and adult lymphoid system in mice with an Ikaros null mutation.

Mice homozygous for an Ikaros null mutation display distinct defects in the development of fetal and adult lymphocytes. Fetal T lymphocytes, and fetal and adult B lymphocytes and their earliest progenitors are absent. Postnatally, hematopoietic stem cells give rise to thymocyte precursors that undergo aberrant differentiation into the CD4 lineage and clonal expansion. The lack of NK cells and some gamma delta T cell subsets and a large reduction in thymic dendritic APCs suggest that Ikaros is essential for establishing early branch points in the postnatal T cell pathway. The lymphoid defects detected in Ikaros null mice reveal critical molecular differences between fetal and postnatal hematopoietic progenitors that dictate their ability to give rise to T cells. These studies also establish Ikaros as a tumor suppressor gene acting during thymocyte differentiation. Phenotypic comparison of this null mutation with a severe dominant-negative Ikaros mutation identifies molecular redundancy in the postnatal hemolymphoid system.

Animals↗
PubMed1996

Spatial organization of the Nim1-Wee1-Cdc2 mitotic control network in Schizosaccharomyces pombe.

In Schizosaccharomyces pombe the onset of mitosis is regulated by a network of protein kinases and phosphatases. The M-phase inducing Cdc2-Cdc13 cyclin-dependent kinase is inhibited by Wee1 tyrosine kinase and activated by Cdc25 phosphatase. Wee1 is negatively regulated by Nim1 protein kinase. Here, we describe investigations aimed at better understanding the role of Nim1 in the mitotic control. The most important finding to emerge from these studies is that Wee1 and Nim1 have different patterns of intracellular localization. Immunofluorescence confocal microscopy has revealed that Nim1 is localized in the cytoplasm, whereas it substrate Wee1 is predominantly localized in the nucleus. Previous studies showed that the Cdc2-Cdc13 complex is located in the nucleus. Diversion of Nim1 to the nucleus, accomplished by addition of the SV40 nuclear localization signal, caused the advancement of M, confirming that Nim1 has restricted access to Wee1 in vivo. We propose that the intracellular distribution of Nim1 and Wee1 may serve to coordinate the regulation of nuclear Cdc2-Cdc13 with cytoplasmic growth.

CDC2 Protein Kinase↗
PubMed1996

Factor V R506Q gene mutation analysis by PCR-RFLP: optimization, comparison with functional testing for resistance to activated protein C, and establishment of cell line controls.

Resistance to activated protein C (APC) has been recently identified as a highly prevalent risk factor for the development of venous thrombosis. In the majority of cases, APC resistance correlates with the presence of a single point mutation in the factor V gene (FV R506Q). The mutation is present in 3% to 5% of the general population and in up to 50% of patients with a personal and family history of venous thrombosis. In the current study, the authors have optimized and implemented for clinical diagnosis a method for detection of FV R506Q using the polymerase chain reaction coupled with restriction fragment length polymorphism analysis (PCR-RFLP). Forty-one healthy adults and 139 patients referred for hypercoagulability testing were genotyped and their APC resistance ratios determined using commercially available reagents (COATEST APC Resistance Kit). Comparative analysis indicated that if functional APC resistance was defined as per manufacturer's guidelines, a significant number of individuals with a normal factor V genotype were categorized as APC resistant and conversely, a significant number of individuals heterozygous for FV R506Q were categorized as non-APC resistant. These results indicate that comparative functional and genotypic analyses in the individual clinical laboratory setting are critical for establishing normal ranges and cut-off values for functional APC resistance due to FV R506Q. To facilitate molecular evaluation of APC resistance, Epstein-Barr virus (EBV) immortalized B-lymphocyte cell lines were established from individuals heterozygous and homozygous for FV R506Q.

Adult↗
PubMed1996

The molecular basis of Boston-type craniosynostosis: the Pro148-->His mutation in the N-terminal arm of the MSX2 homeodomain stabilizes DNA binding without altering nucleotide sequence preferences.

Craniosynostosis, Boston type is an autosomal dominant disorder that results in the premature fusion of calvarial bones and ensuing abnormalities in skull shape. We showed previously that this disorder is tightly linked to the Msx2 homeobox gene on the long arm of chromosome 5, and that affected individuals bear a mutated copy of Msx2. In addition, transgenic mice in which either mutant or wild-type mouse Msx2 is overexpressed in the developing skull also exhibit craniosynostosis. That both mutant and wild-type Msx2 elicit craniosynostosis in transgenic mice and that the Boston type mutation is dominant led us to hypothesize that the mutation might enhance the normal function of Msx2. The mutation is located in position 7 of the N-terminal arm of the homeodomain, a region implicated in both target sequence recognition and protein-protein interactions. Here we test the hypothesis that the Pro148-->His mutation alters the DNA binding properties of Msx2. Using gel shift and binding site selection analyses, we show that the mutation enhances the affinity of Msx2 for a set of known Msx2 target sequences but has little or no effect on the site specificity of Msx2 binding. The enhancement of Msx2 binding is due largely if not entirely to an increased stability of the mutant Msx2-DNA complex. These data provide a molecular-level explanation of how the Pro148-->His mutation enhances Msx2 function and thus leads to the dominant craniosynostosis phenotype.

Animals↗
PubMed1996

Inhibition by cyclic AMP of basal and induced inositol phosphate production in cultured aortic smooth muscle cells from Wistar-Kyoto and spontaneously hypertensive rats.

OBJECTIVE: To investigate inositol phosphate formation and its modulation by the cyclic AMP (cAMP) pathway in cultured aortic smooth muscle cells from spontaneously hypertensive rats (SHR). METHODS: Phenylephrine was used to stimulate inositol phosphate formation in cultured aortic smooth muscle cells from SHR and Wistar-Kyoto (WKY) rats. The smooth muscle cells from passages 6-14 were prelabelled with myo-[2-3H]-inositol (1.9 x 10(5) Bq/ml for 24 h) and inositol phosphate formation was measured after exposure to agonist for 45 min. (-)isoproterenol or forskolin-induced cAMP formation was also evaluated using a radioimmunoassay method. RESULTS: The basal level of inositol phosphate formation in smooth muscle cells from SHR was higher than that observed in smooth muscle cells from WKY rats. Phenylephrine increased the formation of inositol phosphates in a concentration-dependent manner (0.1-100 mumol/l). In the presence of 100 mumol/l phenylephrine, the increase in inositol phosphate formation was significantly greater in smooth muscle cells from SHR (214 +/- 6%) than that observed in smooth muscle cells from WKY rats (156 +/- 8%). When the cells were pretreated with 1 mmol/l 8-bromoadenosine 3':5'-cyclic monophosphate or with 10 mumol/l forskolin for 45 min, the basal production of inositol phosphates in smooth muscle cells both from SHR and from WKY rats was significantly and similarly decreased by about 20%. In the presence of 1 mmol/l 8-bromoadenosine 3':5'-cyclic monophosphate, 100 mumol/l phenylephrine-induced inositol phosphate formation was similarly decreased by 33 +/- 4 and 30 +/- 3% in smooth muscle cells from SHR and from WKY rats, respectively, whereas, in the presence of 10 mumol/l forskolin, inositol phosphate formation was reduced by 25 +/- 3 and 27 +/- 5%, respectively, in those cells. In contrast, isoproterenol induced less inhibition of phenylephrine-induced inositol phosphate formation in smooth muscle cells from SHR (14 +/- 2%) than it did in those from WKY rats (25 +/- 4.5%). Although there was no significant difference in basal or forskolin-induced cAMP accumulation between smooth muscle cells from SHR and those from WKY rats. (-)isoproterenol-induced cAMP accumulation was significantly lower in smooth muscle cells from SHR. CONCLUSION: A marked inhibitory effect of cAMP on the alpha 1-adrenoceptor-mediated inositol phosphate signal transduction pathway was demonstrated in smooth muscle cells of SHR and of WKY rats. Decreased cAMP formation with beta-adrenergic stimulation and increased inositol phosphate formation with alpha-adrenergic stimulation in SHR smooth muscle cells may both contribute to the dominant alpha 1-adrenergic activity observed in SHR.

Animals↗
PubMed1996

Identification of a domain in Rck, a product of the Salmonella typhimurium virulence plasmid, required for both serum resistance and cell invasion.

Rck is encoded on the Salmonella typhimurium virulence plasmid and is a member of a family of related 17- to 19-kDa outer membrane proteins of Enterobacteriaceae, including Ail (Yersinia enterocolitica) and PagC (S. typhimurium). Structural models for these proteins predict eight membrane-spanning domains alternating with hydrophilic inner and outer loops. When expressed in Escherichia coli, Rck and Ail, but not PagC, confer high-level resistance to the bactericidal activity of complement as well as the ability to adhere to and invade mammalian cell lines. To identify functional domains of Rck, we made and screened random mutations in Rck for decreased bioactivity. We found that a single amino acid substitution (glycine to aspartic acid) in the putative third outer loop greatly reduced Rck-mediated serum resistance and eukaryotic cell invasion. We then constructed two chimeric proteins between Rck and PagC. Substitution of the C-terminal half of Rck with the corresponding PagC fragment containing both the third and the fourth outer loops abolishes the Rck-mediated serum resistance and invasion phenotypes. Substitution of Rck with a smaller C-terminal portion of PagC containing the fourth outer loop did not affect the invasive phenotype or serum resistance. These data reveal that the third putative outer membrane loop region is important for the virulence-associated properties of the Rck protein and suggest a similarity between the mechanism of serum resistance and epithelial cell invasion involving the same domain of Rck.

Amino Acid Sequence↗
PubMed1996

Early T lymphocyte progenitors.

The earliest steps along the pathway leading to T cells in mice and humans are reviewed. These are the steps between the multipotent hemopoietic stem cell (HSC) and the fully committed precursors undergoing T cell receptor (TCR) gene rearrangement. At this level significant differences between adult and fetal lymphopoiesis have been demonstrated. The extent of lymphoid commitment of precursors within bone marrow is still unresolved, although HSCs clearly undergo developmental changes before migration to the thymus. Both multipotent and T-restricted precursors have now been isolated from fetal blood, suggesting both may seed the thymus. Within the thymus, several minute but discrete populations of T precursors precede the stage of TCR gene rearrangement. They include precursors that are not exclusively T-lineage committed, although they are distinct from HSCs. These precursors have a potential to form NK cells, B cells, dendritic cells, and sometimes other myeloid cells. Some factors that control early lymphoid development are discussed, including IL-7 and the Ikaros transcription factors. These will eventually help to clarify the process of T-lineage commitment.

Animals↗
PubMed1996

Capillary-to-fiber surface ratio in rat fast-twitch hindlimb muscles after chronic electrical stimulation.

We examined the relative plasticity of capillaries and fiber mitochondria in rat fast-twitch hindlimb muscles in response to chronic electrical stimulation. Specifically we addressed whether the size of the capillary-fiber interface increases in proportion to fiber mitochondrial volume, inasmuch as fiber aerobic capacity increases severalfold with chronic stimulation. Tibialis anterior and extensor digitorum longus muscles of six rats [367 +/- 17 (SD) g body wt] were stimulated (10 Hz, 8 h/day, 7 days/wk) for 28 consecutive days. Subsequently they were perfusion fixed in situ and stimulated, and contralateral control samples from the midbelly were processed for electron microscopy and morphometry. Capillary length density, capillary-to-fiber ratio, and fiber mitochondrial volume density increased two- to threefold in stimulated muscles, with no change in fiber or capillary diameter. Capillary-to-fiber surface area ratio per fiber unit mitochondrial volume was unchanged in stimulated muscles compared with contralateral controls, indicating a proportional increase in the size of the capillary-fiber interface and fiber mitochondrial volume in the muscles after chronic electrical stimulation.

Animals↗
PubMed1996

Molecular genetics and pathophysiology of 17 beta-hydroxysteroid dehydrogenase 3 deficiency.

Autosomal recessive mutations in the 17 beta-hydroxysteroid dehydrogenase 3 gene impair the formation of testosterone in the fetal testis and give rise to genetic males with female external genitalia. Such individuals are usually raised as females, but virilize at the time of expected puberty as the result of increases in serum testosterone. Here we describe mutations in 12 additional subjects/families with this disorder. The 14 mutations characterized to date include 10 missense mutations, 3 splice junction abnormalities, and 1 small deletion that results in a frame shift. Three of these mutations have occurred in more than 1 family. Complementary DNAs incorporating 9 of the 10 missense mutations have been constructed and expressed in reporter cells; 8 of the 9 missense mutations cause almost complete loss of enzymatic activity. In 2 subjects with loss of function, missense mutations testosterone levels in testicular venous blood were very low. Considered together, these findings strongly suggest that the common mechanism for testosterone formation in postpubertal subjects with this disorder is the conversion of circulating androstenedione to testosterone by one or more of the unaffected 17 beta-hydroxysteroid dehydrogenase isoenzymes.

17-Hydroxysteroid Dehydrogenases↗
PubMed1996

[The effect of mitomycin C on filtration surgery of glaucoma with poor prognosis].

OBJECTIVE: To determine the anticicatrization effect of mytomycin C (MMC). METHODS: We randomly divided 30 cases (40 eyes) into two groups: 21 eyes in MMC group and 19 eyes in control group. Intra-operatively, 0.4 mg/ml MMC was used in the trabeculectomy in MMC group, and no MMC was used in the control group. The post-operative follow-up periods ranged from 6 to 25 months (mean, 10.0 months). RESULTS: The successful rate of the operation of MMC group was 90.4% and that of the control group, 26.3% (P < 0.0001). The rate of eyes with functional filtering blebs was 17/21 and that of the control group, 4/19 (P = 0.002). Macular edema occurred in 3 eyes of MMC group and none in the eyes in the control group. There were no corneal complication and leakage of the wound. CONCLUSION: The results show that MMC can promote the formation of functional filtering bleb and elevate the successful rate of filtration surgery.

Adolescent↗
PubMed1996

[Effect of antisense c-erbB2 on biologic behaviour and chemotherapeutic drug sensitivity in human ovarian cancer cells].

OBJECTIVE: To explore the effects of antisense erbB2 on the biological behaviours and on chemotherapeutic drug sensitivity in human ovarian cancer cells. METHODS: A recombinant retroviral expression vector: pDOR-erbB-neo was constructed which contain the neomycin resistant gene and the erbB2 cDNA 3.8kb antisense fragment. pDOR-erbB-neo was introduced into human ovarian cancer cell line SKOV3 by lipofectin. The cells were selected with geneticin (G418). The cells transfected with pDOR-erbB-neo were named SKOV3-A2. RESULTS: Southern blot analysis confirmed the presence of the antisense erbB2 in transfected cells. Transfected cells showed no obvious changes in morphology. Compared with parental cells and with pDOR-neo transfected cells used as controls, the growth and DNA synthesis of antisense transfected cells, SKOV3-A2, were inhibited. Transfection with pDOR-erbB-neo rendered the cells significantly, more sensitive to chemotherapeutic drugs (5-fluorouracil, cisplatinum) than the parental cells. CONCLUSIONS: The effectiveness and the potentiality of antisense erbB2 in ovarian cancer gene therapy is demonstrated. c-erbB2 oncogene may be related to drug resistance in ovarian cancer cell.

Antineoplastic Agents↗
PubMed1996

Further characterization of the metabolic properties of triglyceride-rich lipoproteins from human and mouse apoC-III transgenic mice.

We previously showed that human apoC-III expression in transgenic mice causes hypertriglyceridemia due to the accumulation of enlarged very low density lipoprotein (VLDL)-like particles, with increased triglycerides and apoC-III and decreased apoE. In vivo turnover studies indicated the metabolic basis was decreased particle fractional catabolic rate. The presence of enlarged triglyceride-rich particles with prolonged residence time in plasma implied defective lipolysis, but in vitro these particles were good substrates for purified lipoprotein lipase (LPL). In the current study we further characterize the metabolic properties of these particles. We show that expression of a mouse apoC-III transgene can also cause hypertriglyceridemia with a similar accumulation of a VLDL-like particle with increased apoC-III and decreased apoE. A vitamin A fat tolerance test was used to show that MoCIIITg and HuCIIITg mice had similarly delayed clearance of triglyceride-rich postprandial particles. Thus, the previously observed hypertriglyceridemia caused by human apoC-III transgene expression was not due interspecies incompatibility but a property of apoC-III. In further experiments we showed VLDL from apoC-III transgenic mice interacted poorly with fibroblast lipoprotein receptors and this could be corrected by adding exogenous apoE. In addition, control VLDL interaction could be decreased by exogenous apoC-III. Moreover, the hypertriglyceridemia of HuCIIITg mice could be normalized by crossbreeding with HuETg mice. Thus, a functionally significant reciprocal relationship of apoC-III and apoE exists, presumably due to competition for space on the surface of triglyceride-rich lipoproteins. Finally, VLDL from HuCIITg and MoCIIITg mice showed decreased binding to heparin-Sepharose. This suggests and additional locus of the defect in these mice could potentially be in the binding of triglyceride-rich lipoproteins to heparan sulfate proteoglycan matrix on the surface of endothelial cells in which LPL is embedded. This could explain the predicted functional lipase deficiency in apoC-III transgenic mice based on the observation of a prolonged residence time of enlarged triglyceride-rich lipoproteins.

Animals↗
PubMed1996

Studies on prophylactic effect of artesunate on schistosomiasis japonica.

OBJECTIVE: To examine the prophylactic effect of artesunate on schistosomiasis japonica by killing schistosomula. METHODS: Mice, rabbits and dogs after infection with cercariae of schistosoma japonicum (S. japonicum) were treated with artesunate on the 7th day at a dose of 300 mg/kg, 20-40 mg/kg and 30 mg/kg respectively once a week for 4-6 weeks. A double-blind test was used. A total of 864 persons in highly endemic areas for schistosomiasis were administered either artesunate at a dose of 6 mg/kg once a week for 8 weeks or identical placebo with the same dose-schedule during transmission season for S. japonicum. Four weeks after the last dosing, fecal examinations for S. japonicum eggs and miracidia were carried out to evaluate the prophylactic effect. RESULTS: Worm reduction rates in mice, rabbits and dogs were 77.50-90.66%, 99.53% and 97.10% respectively. All of the 467 residents in 2 trials were free from eggs or miracidia upon stool examinations whereas in the control groups with placebo, 15 out of 218 (6.9%) and 26 out of 179 (14.5%) were stool eggs and/or miracidia positive in the first and second trial, respectively. Side effects were mild. No significant changes in routine blood and urine tests, electrocardiogram, hepatic and renal functions were observed after artesunate administration. CONCLUSIONS: Animal experiments and field trials have demonstrated good efficacy of artesunate on killing schistosomula with little side effects. Thus, the drug is suggested to be used on the population in endemic areas for prevention of schistosomiasis.

Adolescent↗
PubMed1996

Role of intracardiac angiotensin II in cardiac dysfunction of rat during septic shock.

OBJECTIVE: To observe changes in the content of myocardial angiotensin II and its role in cardiac dysfunction of rat during septic shock. METHODS: Septic shock model was produced by cecal ligation and puncture (CLP) operation on rats. Experimental rats were given captopril 15 mg.kg-1/d per os for 3 days before CLP operation. Mean blood pressure and left ventricular pressure were recorded. Myocardial angiotensin converting enzyme (ACE) activities were determined by a fluorometric assay and myocardial angiotensin II content was measured by radioimmunoassay. Highly purified membrane of sarcoplasmic reticulum (SR) was prepared from rat hearts. Assays were made of ATP-dependent Ca2+ uptake by cardiac SR and (3H) ryanodine binding to SR. RESULTS: Myocardial angiotensin II content increased by 51.5% (P < 0.01) at the 18th hour post CLP, meanwhile there was a decrease in left ventricular +/- dp/dtmax value and the impairment in Ca2+ uptake and (3H) ryanodine binding to cardiac SR. Preliminary administration of captopril reduced myocardial ACE activity and angiotensin II content, but increased left ventricular +/- dp/dtmax value. In comparison to shock group, the initial rate and the capacity of SR Ca2+ uptake were increased by 120% (P < 0.01) and 33.9% (P < 0.05), the Bmax value of (3H) ryanodine binding to SR was also elevated, while the Kd value remained unchanged. CONCLUSIONS: The elevated intracardiac angiotensin II, resulting from the activation of myocardial ACE during sepsis, probably serves as one of the important mediators participating in the pathogenesis of heart failure: the effects of angiotensin II may be associated with the disturbance of Ca2+ transport function of cardiac SR.

Angiotensin II↗
PubMed1996

[A comparative study on coronary angiography and exercise ECG testing].

Both coronary angiography and exercise ECG testing were done in 461 patients. The comparative study showed that positive results of exercise ECG testing down-slope or horizontal depression of ST segment more than 0.1 mV during or after exercise may predict coronary artery disease, the sensitivity was 74.5% (322 cases) and specificity 84.2% (139 cases). With the use of ST depression more than 0.2 mV as the super-positive standard, the sensitivity and specificity of positive exercise ECG testing to predict triple vessel disease and left main coronary disease was 61.3% and 79.1%, respectively. The workload and the increment of systolic blood pressure had a negative correlation (r = -0.4531 and -0.3), but the duration of ST depression had a positive correlation (r = 0.4502) with the number of diseased coronary arteries. The patients with multi-vessel disease and left main coronary artery disease had a higher positive rate of exercise ECG testing as compared with the patients with single or double vessel disease. The female patients had a higher false positive rate than the male. It is concluded that exercise ECG testing has some value in diagnosing coronary artery disease and predicting its severity. Some parameters in exercise is helpful to predict the severity of coronary artery disease.

Adult↗
PubMed1996

[Clinical study on painless labor under drugs combined with acupuncture analgesia].

To study the analgesia efficacy of drugs combined with acupuncture analgesia for painless labor, 462 normal pregnancy women were observed. During the latent phase in labor, several analgesia methods: acupuncture analgesia, analgesics, magnetotherapy and auricular acupressure, TENS combined with dihydroetorphine were used respectively. While the intrauterine pressure and the peripheral content of beta-EP were measured during labor, the experiments of SEPS were also performed on healthy adults to demonstrate the analgesia efficacy of those analgesia methods. The combination of drugs with acupuncture is an excellent method for painless labor without any complications and all the mothers and babies are safety. The effectiveness is 97.5%. The results demonstrate that the mechanism of analgesia efficacy should be regulated the incoordinate uterine action and improve the hypertonic status of uterus, but also can decrease the pain threshold and elevate the tolerance of uterine contractions during labor.

Acupressure↗
PubMed1996

156 cases of Gilles de la Tourette's syndrome treated by acupuncture.

The principle of clearing Yangming and nourishing the kidney and heart was adopted in the treatment of 156 cases of Gilles de la Tourette's Syndrome with acupuncture. The total effective rate was 92.3%, and the cure rate in children aged 11-15 years was markedly higher than that in children 6-10 years of age. Among 84 cases with abnormal EEG, the pathological waves in 54 disappeared or ameliorated after the treatment.

Acupuncture Therapy↗