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Biomedical subjects

Z H Li

Publications and source records attributed to Z H Li.

At least 19 recordsLinked to original sources

Cross-linking of surface immunoglobulin activates src-related tyrosine kinases in WEHI 231 cells.

Crosslinking of sIgM on the B cell line WEHI 231 with anti-sIgM antibody induces protein tyrosine phosphorylation, implicating protein tyrosine kinases (PTKs) in sIg-mediated signal transduction. We have analyzed this cell line for members of the src family of PTKs and have evaluated whether these PTKs might be involved in the process of sIgM-mediated signaling. Our results show that Blk, Lyn, Lck, and Hck are detectable in WEHI 231 cells. Addition of antibodies to sIgM were found to variably stimulate the activities of Blk, Lyn, Lck, and Hck as measured by immune-complex protein kinase assays. Autophosphorylation of these src PTKs, as assessed by reaction with anti-phosphotyrosine antibodies, increased over the time course of sIgM-mediated activation. Co-immunoprecipitation studies to investigate the potential physical interaction of src PTKs with the sIgM receptor complex revealed that, under digitonin and Brij 96 lysis conditions Lyn, Lck, Hck, but not Blk associated with sIgM.

Animals

Ecology and prevention of a shellfish-associated hepatitis A epidemic in Shanghai, China.

During a shellfish-borne hepatitis A outbreak in Shanghai during the first quarter of 1988, 300,000 cases were reported in two months. Using cell culture and experimental infection of marmosets, hepatitis A virus (HAV) was isolated from clams collected from the market and the sea bed during the epidemic. A dose-response curve correlating the quantity of clams consumed to the attack rate of hepatitis A was well documented. The occurrence of the epidemic was associated with a good harvest of clams in a new area, serious pollution of this area with sewage and importation of the clams in large quantities into Shanghai where most young adults were susceptible. Clams can apparently be decontaminated by using a continuous water flow. In this way, HAV titres can be reduced by 90% in one day and by 99.9% in two weeks. An attenuated live HAV vaccine which has been developed in China has been shown to be safe and immunogenic and may be used for prevention of such epidemics in the future.

Adolescent

Adipocytes of old rats produce a decreased amount of differentiation factor for preadipocytes derived from adipose tissue islets.

Adipose tissue of young rats (3.5 mo) contains cell formations ("islets") that, in culture, give rise to a variety of preadipocytes, or islet-derived cells (IDCs), that undergo rapid morphologic differentiation into adipocytes. Such adipose conversion depends on a differentiation-promoting factor produced by the mature adipocytes also present in the cultures. Here we report that cultured IDCs from epididymal fat of senescent rats (20 +/- 3 mo) show decreased adipose conversion compared to IDCs from 3.5 mo rats at 3 days of culture (14.2% vs 29.9%; p < .001). Both the number of co-cultured adipocytes and increased fragility with age were excluded as having a substantial role in explaining the decreased conversion. In order to determine whether the decrease differentiation of the IDCs of the old rats was intrinsic or due to reduced production of the differentiation factor by the old adipocytes, cross-over cultures of IDCs from young and old rats were grown in the presence of young or old adipocytes. When IDCs of old rats were cultured in the presence of young adipocytes, a significant twofold increase in differentiation was seen compared to old IDCs grown with old adipocytes (22.7% vs 10.3%; p < .001). This response was comparable to that of young IDCs grown with young adipocytes. Thus, old IDCs retain their ability to undergo extensive morphologic differentiation when appropriately stimulated. Reduced production of the differentiation-promoting factor by old adipocytes rather than the ability of IDCs to differentiate appears to be responsible for the decreased adipose conversion of IDCs seen in cultures of adipose tissue from senescent rats.

Adipose Tissue

Specific genetic analysis of microscopic tissue after selective ultraviolet radiation fractionation and the polymerase chain reaction.

A method using selective ultraviolet radiation fractionation followed by polymerase chain reaction (PCR) can analyze specific cell subsets present on a microscope section. Direct ultraviolet radiation of fixed and stained tissue sections prevents subsequent amplification by PCR. An "umbrella" or dot placed physically over small numbers of pure cell populations selected by microscopic examination protects these cells from the ultraviolet inactivation. The DNA in these protected cells can be specifically amplified while no signal is derived from the unprotected surrounding cells. Specific amplification was demonstrated by detecting human papillomavirus sequences only if infected cells were protected. Similarly, loss of heterozygosity at the p53 locus was documented by selective dotting of normal or tumor cells. The method allows the specific and sensitive molecular genetic analysis of small numbers of cells histologically identified and selected under the microscope.

Base Sequence

Analysis of styryl-based inhibitors of the lymphocyte tyrosine protein kinase p56lck.

Several styryl-based compounds were evaluated for their capacity to act as inhibitors of the non-receptor tyrosine protein kinase p56lck. Our results demonstrate that alpha-cyanocinnamamide compounds can inhibit both the in vitro tyrosine autophosphorylation of p56lck as well as p56lck phosphorylation of exogenous substrates. Compound 67B-83-A was found to inhibit p56lck protein kinase activity with a calculated IC50 of 7 to 10 microM. This compound did not significantly inhibit the tyrosine protein kinase activity of the epidermal growth factor receptor and was found to be a less effective tyrosine protein kinase inhibitor for other members of the src family of protein kinases.

Animals

Islets of preadipocytes highly committed to differentiation in cultures of adherent rat adipocytes. Light- and electron-microscopic observations.

Cultures of adherent mature adipocytes, obtained from collagenase-digests of adipose tissue of the rat, invariably contain rapidly proliferating, fibroblast-like cells despite the washing and centrifugation procedures employed during isolation of the fat cells. Such spindle-like cells originate from low-density structures, which we term "islets", that are present, together with the mature adipocytes, in the floating layer of the digest of adipose tissue. Islets are found in preparations from adult (3-4 months old) as well as aging (17-24 months old) rats. By light- and electron microscopy, the islets appear as clusters of closely associated cells containing a variable amount of lipid-like material. Cells of endothelial or pericytic origin are also present in the islets. Within a few hours of culture, the islets give rise to those spindle-like cells that have been seen to proliferate in the cultures. By 36-48 hours, such cells begin to accumulate lipid droplets and, by 150 hours, assume the morphology of small mature adipocytes (diameter 20-35 microns) with a large central lipid droplet. The pattern of differentiation of these cells recalls that of preadipocytes derived from the stromal-vascular fraction of adipose tissue digests. Nonetheless, the extent and rapidity of their adipose conversion, as well as the culture conditions necessary for differentiation, are different and suggest that these cells are a substantially uniform subpopulation of adipocyte-precursor cells highly committed to differentiation.

Adipose Tissue

Phosphonate-containing inhibitors of tyrosine-specific protein kinases.

Tyrosine-specific protein kinases (TPK) are important signal transducing enzymes involved in normal cellular growth and differentiation and have been implicated in the etiology of a number of human neoplastic processes. Efforts to develop agents which inhibit the function of these enzymes by interfering with the binding of substrate have been limited by the lack of detailed three-dimensional structural data. Many inhibitors of substrate binding share a common styrene nucleus 1 which has been postulated to function as a conformationally constrained analogue of tyrosine. In an effort to develop high-affinity compounds based on this hypothesis, a number of derivatives were synthesized in which either methylphosphonate (4a-c) or (hydroxymethyl)phosphonate (3a-c) were appended to the aromatic 4-position of styrene-containing moieties. The intent of this approach was to prepare hydrolytically stable analogues which expressed additional enzyme recognition features present during the phosphorylation of tyrosine itself. None of the analogues showed inhibitory activity up to the maximum concentration tested (1000 microM) when assayed against autophosphorylation of A-431-derived epidermal growth factor receptor (EGFR) or p56lck (autophosphorylation and transphosphorylation of rabbit muscle enolase). Additionally, a series of naphthalene-based inhibitors including (1-naphthalenylhydroxymethyl)phosphonic acid (14), its known 2-positional isomer 16, and sulfonate (19, 20) and phosphate derivatives (17, 18) were also tested under similar conditions. Only (2-naphthalenyl-hydroxymethyl)phosphonic acid (16) showed activity (IC50 = 250 microM in EGFR, in agreement with the reported literature value). These results suggest that the interaction of styrene-based inhibitors with the substrate binding domain of TPKS may not occur in a manner analogous to the interaction of tyrosine with this domain.

Animals

[Determination of elastase and alpha 2-macroglobulin in bronchoalveolar lavage fluid in patients with interstitial diseases and its clinical significance].

Determination of elastase and its inhibitor alpha 2-Macroglobulin was done in BALF in 40 patients with ILD (study group) and 10 healthy volunteers (control group). The results showed that BALF levels of elastase and alpha 2-macroglobulin in study group were significantly higher than those in control group (P less than 0.01; P less than 0.05). Correlation was found between BALF elastase levels and cell count (r = 0.3214, P less than 0.05). It was suggested that elastase may play a role in the development of pulmonary fibrosis in interstitial lung diseases.

Adult

Dietary alterations of plasma lipoproteins influence their interactions with proteoglycan enriched extracts from neointima of normal and injured aorta of rabbit.

The interactions between serum lipoproteins isolated from rabbits fed a cholesterol-supplemented diet for six weeks, and soluble extracts of arterial neointima enriched in proteoglycans extracted from normocholesterolaemic rabbit aortas, were studied in an in vitro system. Neointimal tissues of rabbit aorta, which developed during three months following a selective endothelial injury, were excised and the areas covered or uncovered by regenerated endothelium were separated. To isolate the proteoglycan enriched fraction, both normal and injured tissue was homogenized in a sucrose solution containing protease inhibitors, centrifuged, and further fractionated by gel exclusion chromatography. The composition of the soluble extracts and each of their corresponding proteoglycan enriched fractions were analyzed in terms of protein and glycosaminoglycan content. Lipoproteins of donor animals fed an atherogenic diet were prepared by sequential ultracentrifugal flotation after density adjustment with KBr. Aliquots of electrophoretically pure lipoprotein fractions were incubated with proteoglycan enriched fraction from uninjured, denuded, or endothelium-covered neointima in the presence of Ca++ and Mg++ at 4 degrees C. The complexes formed during incubation were separated by centrifugation. The cholesterol content of the complexes was considered as an index of binding capacity. Results were expressed as micrograms of cholesterol bound per mg of glycosaminoglycan. The data reveal the higher affinity of hypercholesterolaemic lipoprotein fractions for aortic proteoglycans, as compared to normocholesterolaemic lipoproteins. In addition, when evaluating the relevance of the proteoglycan enriched fraction source, the affinity of fractions extracted from aortic neointima was found to be much higher for hypercholesterolaemic lipoproteins. These results suggest the role that proteoglycan-lipoprotein interactions could play in the event of the combined actions of endothelial injury and hypercholesterolaemia in the pathogenesis of atherosclerosis.

Animals

[Distribution of lectin-receptors in normal, dysplastic and neoplastic cervical epithelium].

112 cases of normal, dysplastic and neoplastic cervical epithelium were studied with a panel of twelve various lectins and ABC technique. The results showed that: (1) ConA and WGA receptors were relative to Squamous epithelial origin of the Cervix. (2) PNA, UEA-1, BSL and PHA receptors correlated with the tumorigenicity of cervical squamous epithelium. (3) WGA receptor correlated with the cell differentiation of squamous carcinomas. (4) DBA receptor was related with tumor invasion. (5) ConA and SJA receptors were related to the tumorigenicity of columnar epithelium of the endocervical glands. (6) Applications of neuraminidase caused compositional changes of glycoconjugates in the receptors of normal, dysplastic and malignant cervical epithelium and this may be of some value in clinical practice.

Adenocarcinoma

[Studies on the influence of air cavities during irradiation with high energy electrons].

Radiation therapy of tumors is often performed with high energy electrons. In certain situations air cavities in the irradiation volume can change the dose distribution. The effects of air cavities on the dose distribution are investigated. The results calculated with a therapy planning system and measured in a phantom are described and compared with one another and with the literature.

Humans

On the choice of times for data analysis in group sequential clinical trials.

Planned interim analysis of randomized clinical trials has been implemented for over a decade. While the initial proposal advocated analyzing after equal numbers of patients were evaluated, a later modification by Lan and DeMets (1983, Biometrika 70, 659-663) allowed for more flexible boundaries. Rather than fixing the times of analysis at equal numbers of patients, they fixed the rate at which overall alpha was used up according to a use function alpha * (t) on t in with alpha * (0) = 0 and alpha * (1) = alpha. Here we consider how flexible Lan and DeMets' procedure is. We show that the choice of alpha * (t) for a particular trial affects the permissible analysis times if other desirable properties of the sequence of nominal significance levels are to hold. To overcome the difficulties posed by patterns of late analysis, piecewise linear convex use functions are proposed.

Biometry

Adipose tissue islets: tissue culture of a potential source of fat cells in the adult rat.

Collagenase digests of adipose tissue of the 3 to 4-month-old rat contain groups of 20-100 tightly arranged cells (islets) that copurify with the free-floating fat cells. When cultured along with mature adipocytes the islets give rise to cells, initially fibroblast-like, which rapidly proliferate, acquire lipid droplets, and differentiate into small adipocytes within 4-6 days without the addition to the medium of the agents usually required to produce differentiation in stromal-vascular preadipocytes. Differentiation of these cells is independent of confluence and begins as early as day 2 of culture. The proportion of islet-derived cells that differentiate is directly correlated with the number of mature adipocytes simultaneously present in the culture (r = .709; P less than 0.001). Culture medium exposed to mature adipocytes demonstrated differentiation-promoting activity, suggesting a paracrine effect of these cells. Islets may in vivo constitute a source for newly formed adipocytes in the adult rat. The differentiation of these potential adipocytes may be regulated, at least in part, by the mature fat cells via a paracrine effect.

Adipose Tissue

X-ray diffraction studies on the absolute configuration of alpha- and beta-anordrins.

The molecular structures and absolute configurations of alpha- and beta-anordrins are reported. Pure alpha- and beta-epimers were obtained with recrystallization and column chromatography combined with high-pressure liquid chromatography methods; they were identified by high-resolution infrared and mass spectra and 1H and 13C nuclear magnetic resonance. By single crystal x-ray diffraction analysis, the crystals of alpha- and beta-epimers were found to belong to the orthorhombic space groups P2(1)2(1)2(1) and P2(1)2(1)2, respectively. The molecular structures of these two epimers were determined. The absolute configurations were deduced by conformation analysis, 1H nuclear magnetic resonance, and comparison with the absolute configuration of the starting material. The absolute configurations of asymmetric centers of alpha- and beta-epimers were observed to be 2R, 5S, 8R, 9S, 10S, 13S, 14S, 17R, and 2S, 5S, 8R, 9S, 10S, 13S, 14S, 17R, respectively. These results were confirmed by the x-ray diffraction determination of the absolute configuration of 2 alpha,17 alpha-diethynyl-A-nor-5 alpha- androstane-2 beta, 17 beta-diol dichloroacetate.

Chemical Phenomena

[Relationship between the positive inotropic action of matrine and extracellular calcium].

Matrine (Ma) was extracted from the seed and leaves of Sophora alopecuroides L. Ma showed positive inotropic actions in isolated guinea pig left atrium and rat vas deferens under electric stimulation. The positive inotropic actions were markedly inhibited by verapamil (1 mumol/L). Isolated rat vas deferens under the action of Ma showed rhythmic contraction. High K+ (KCl 40 mmol/L) augmented the frequency of rhythmic contraction induced by Ma; conversely, Ma may also augment the increased tension produced by high K+. These results suggest that, the positive inotropic actions of Ma is relevant to its activation of Ca2+ channels.

Alkaloids

[X-ray diffraction studies on the configuration of 2 xi, 17 alpha-diethynyl-2 xi, 17 beta-dihydroxy-A-nor-5 alpha-androstane alpha-epimer].

X-ray diffraction studies on the configuration of 2 xi, 17 alpha-diethynyl-2 xi, 17 beta-dihydroxy-A-nor-5 alpha-androstane "alpha-epimer" that possessed antifertility effect and estrogen activity were reported. Pure alpha-epimer was obtained by recrystallization and low pressure silica gel column chromatography combined with HPLC method. Its structure was identified by IR, MS, 1HNMR. The configuration was determined by single crystal X-ray diffraction analysis. The crystal of alpha-epimer belonged to orthorhombic, the space group was P222(1), with the following crystallographic parameters: a = 6.777 (2), b = 12.125 (4), c = 25.292 (8)A, V = 2078.5(1.2)A3, Z = 4. One thousand two hundred and thirty-five independent reflections with I greater than or equal to 3 sigma (I) were collected on a Nicolet R3M/E four-circle diffractometer by means of MoK alpha radiation. The structure was solved by direct method and refined by least square technique to a final discrepancy factor of R = 0.039. The molecule was shown to consist of the alpha-configuration of C2 and C17ethynyl groups. The absolute configuration was deduced by the absolute configuration of synthetic raw material, conformation analysis and the study of 1HNMR. The absolute configuration of asymmetric centers were 2R,5S,8R,9S,10S,13S,14S,17R.

Crystallization