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Biomedical subjects

Q L Zhu

Publications and source records attributed to Q L Zhu.

At least 19 recordsLinked to original sources

Acid helix-turn activator motif.

A common sequence/structural motif pattern has been identified within the steroid/thyroid hormone receptors and other transcriptional activators using a new massively parallel symbolic learning assistant computer system. The pattern appears nearly diagnostic of transcription activation, including relative activation strength, among nuclear and DNA-binding prokaryotic proteins. In cases where mutation/deletion/chimeric studies have identified the activation domain, the pattern matches within that domain. These facts and the nature of the pattern itself strongly support the idea that the patterned domain is directly involved in a protein-protein transcription activation interaction.

Amino Acid Sequence

Redistribution of fibronectin and keratin localization patterns in early wounded confluent PtK2 cells.

Single and double-label immunofluorescence were used to study the fibronectin (FN) and keratins (Ks) localization patterns in early wounded confluent PtK2 cells. A time-course study (0 hr, 2 hr, 6 hr and 24 hr) gives the following results: before wounding, the FN localizations of confluent cells are composed of curved and sometimes branched strands or fibrils. The Ks network is formed by radial fluorescent filaments connecting the Ks centers near the nuclei with a linear fluorescence underlying the cell membrane. Two hr after, the FN localizations are redistributed at the cell-cell contact areas. The radial Ks filaments are compacted around the nuclei, some of them delineate the cytoplasmic periphery of the wounded cells. Six hr later, the method shows redistributed FN localizations at the cell-cell contact areas. An alveolar pattern is formed enclosing each of the adjacent cells. The codetected Ks filaments are retracted around the nuclei. The underlying cell-cell contact areas are also well demonstrated. It may be noted that these areas are FN-labelled. Twenty-four hr after wounding, the FN alveolar pattern persists. The redistributed Ks filaments have some similarity to those seen before wounding.

Animals

Fibronectin network recovery in confluent PtK2 cells after acrylamide treatment.

Confluent PtK2 cells 4 hr treated with 5 mM acrylamide were FN-detected by indirect immunofluorescence. The initial fibrillar-FN network was replaced by an alveolar-type network located at the cell-cell contacts areas in the form of a thick frame with a lace-like appearance. Afterwards, acrylamide removal was obtained by several washes with fresh FCS-free culture medium. Then, PtK2 cells were returned to the incubator for 20 hr. Cell recovery was indicated by reversion of the initial fibrillar-FN network. These data show that FN reversion was possible without any changes in shape and cytoplasmic organization of non-motile growing cells.

Acrylamide

Culture media conditioned by wounded cells modify the fibronectin localization pattern of unwounded confluent PtK2 cells.

A confluent PtK2 cell sheet was incised in a serum-free culture medium, at 15 min, 2 hr and 24 hr after wounding. The culture media were collected in the same way and used as conditioned media. Unwounded confluent cells were cultured in the conditioned medium for 24 hr. They showed a modification of fibronectin localization similar to that which we had previously observed in wounded confluent PtK2 cells: cells lost their normal fibronectin fibrils and were surrounded by fibronectin lace. This finding suggested that during wound healing, the cells released soluble chemical factors which could modify the fibronectin localization pattern of unwounded confluent cells. Subconfluent cells did not respond to conditioned media, showing that confluent cells and subconfluent cells had different susceptibilities.

Animals

Anti-fibronectin serum inhibits the disorganization of the dermal-epidermal junction in cultured wounded skin.

During wound-healing in cultured frog skin fragments, fibronectin (FN) was detected in the dermal-epidermal junction. Intracellular fibronectin was stained using permeabilization and DAB immunoperoxidase. With electron microscopy intracytoplasmic FN granules were localized in the epidermal processes of the stratum germinativum cells protruding towards the dermis and in their marginal regions (membrane-associated plaques). Faint staining was visible at the level of the lamina densa and inside some parts of the lamina lucida. In comparison, contrasted ultrathin sections revealed classical disorganization of the dermal-epidermal junction. In the presence of anti-fibronectin serum during the whole time of culture, fibronectin-antifibronectin binding was visualized in the form of sparse cytoplasmic granules in the epidermal processes of the stratum germinativum cells. Contrasted ultrathin sections emphasized the continuity between the tonofilaments, the anchoring filaments and the anchoring fibrils. Briefly, anti-fibronectin serum inhibits the disorganization of the dermal-epidermal junction in cultured wounded skin.

Animals

A prospective, controlled, double-blind, cross-over study of tripterygium wilfodii hook F in treatment of rheumatoid arthritis.

A polyglycosides of Tripterygium wilfodii hook F (TWH) preparation with a code name of T2 is used in the present double-blind, controlled, cross-over study on the treatment of 70 cases of rheumatoid arthritis (RA). An impressive curative effect of T2 is confirmed much more convincingly by the present study than that by the previously reported clinical open trials. The adverse reactions and the probable pharmacological mechanism of T2 are also discussed.

Adult

Studies on the structure of HBV DNA.

The structure of HBV adr NC-1 DNA is analyzed and compared with another five strains of HBV DNAs. Some of the prokaryotic promoter-like sequences, palindrom sequences and ATAA are found. An enhancer core sequence and some other characteristics are also shown. In considering the frame and its regulatory sequence as a transcriptional unit some of the possible new frames are discussed.

Base Sequence

Wheat germ agglutinin (WGA) reduces ADH-induced water flow and induces cell surface changes in epithelial cells of frog urinary bladder.

The functional and structural changes induced by apical wheat germ agglutinin (WGA) 100 micrograms/ml exposure on frog urinary bladder have been investigated and the possible correlations between these effects discussed. Bladders, apically exposed to WGA for 30 min to 3 hr exhibit a marked reduction of their response to antidiuretic hormone (ADH) challenge and of their hydrosmotic reactivity. Structural changes triggered by WGA treatment are: 1. apical invaginations of the plasma membrane, interpreted as endocytotic in nature, taking into account the results of carbohydrate cytochemical detection and horseradish peroxidase (HRP) exposure: 2. cytoskeleton disorganization and microvilli collapse. These phenomena do not interfere with cortical granule traffic and are independent of ADH challenge: they occur in ADH-stimulated bladders as well as in bladders at rest. These findings could be interpreted as follows: binding of the divalent lectin WGA to its coat specific receptors would induce changes in the apical membrane structure which in turn could provoke disorganization and disruption of apical cytoskeletal elements associated with plasma membrane. Reduction of bladder response to ADH challenge could result from a reduced recycling of aggrephores, as they are associated with cytoskeletal elements in the subapical cytoplasm. Collapse of microvilli and endocytotic events also could result from apical cytoskeleton disruption, as microvilli are sustained by bundles of actin filaments interconnected with apical cytoskeletal filaments and as plasma membrane is associated with apical cytoskeleton. However, these two last events evidently occur in ADH-challenged or non-challenged bladders.

8-Bromo Cyclic Adenosine Monophosphate

A method of isolation of apical membranous sheets from frog urinary bladder epithelium by stripping with gelatin.

We have developed a technique for recovering apical membranous sheets from amphibian urinary bladders by gelatin stripping. The tissue is mounted on a lucite support and the apical surface is first stuck onto a gelatin-coated glass slide at 30 degrees C. This sandwich is then chilled on ice and the bladder is pulled away from the slide. Preliminary results indicate that this simple technique could be used to remove membranous apical sheets of various sizes, almost devoid of cytoplasmic contamination and without significant damage to the underlying cell structures. The method could also be adapted to prepare perforated cells and to study the cohesive forces between the different layers of the tissue.

Animals

Skin cultured with or without an NaCl gradient and aldosterone influence epidermal fibronectin localizations.

Fibronectin (FN) localizations in the epidermal cells of the frog Rana esculenta were detected in isolated ventral skin fragments 4 day-cultured with or without an NaCl transepithelial gradient and aldosterone. Without the gradient, few mitochondria-rich cells (MRCs) were FN-detected. Stratum germinativum and spinosum cells also contained fibronectin. With the gradient, numerous MRCs were detected. Below them, in the stratum germinativum, clear spaces were recognized. Aldosterone with or without the gradient modified the above effects: in both cases, many MRC contained fibronectin. It was interesting to note that, for each type of culture, stratum germinativum cells were dramatically FN-detected.

Aldosterone

An immunocytochemical method for the localization of fibronectin in Araldite-embedded specimens.

The detection of fibronectin (FN) in osmium-fixed and Araldite-embedded frog skin fragments was studied using a modification of Baskin's procedure (Baskin et al. 1979). Following the removal of Araldite from the semi-thin sections (0.5-1.0 micron) with ethanol-NaOH solution, the sections were bleached with hydrogen peroxide. FN was detected by indirect immunoperoxidase method. For precise localization of FN, careful attention was paid to the temperature, antibody concentrations and the quality of the ethanol-NaOH solution. Our results were in agreement with those that we had obtained previously for polyethylene glycol (PEG) sections, suggesting that the present procedure is useful for the detection of FN in Araldite-embedded biological specimens.

Animals

Cultured epidermis influences the fibril organization of purified type I collagen gels.

Purified type I collagen gel used as culture substrate was composed of unstriated fibrils. Before culture, gel fragments were coated with culture medium with or without fetal calf serum (FCS+ coated or FCS- coated gels). Each gel fragment was apposed to a fragment of frog skin at the medium/air interface in Trowell culture chamber. After 7 days at 20 degrees C, the coated gels were covered with newly formed epidermis containing fibronectin localized around the keratinocytes, whose morphology was considerably modified. Fibroblast-shaped keratinocytes were localized in the anterior zone of the newly formed epidermis on FCS+ gels. The long axis of the cells was parallel to the gel surface, where numerous unstriated fibrils were located. Polyhedral keratinocytes were located in the posterior zone on FCS+ gels or the anterior and posterior zones on FCS- gels with the long axis perpendicular to the gel surface. Numerous cross-striated fibrils were found under the cultured keratinocytes in the vicinity of the basal filipodia. This model is useful for the study of collagen gel reorganization by keratinocytes.

Animals

Fibronectin (FN) localizations in mitochondria-rich cells of frog epidermis.

Indirect immunoperoxidase detection of fibronectin (FN) in the ventral frog epidermis showed that only one type of epidermal cell, the mitochondria-rich cells (MRC), was FN-detected. FN was found in MRC having rounded, flask-like and intermediate shapes, located at different levels of the epidermis between the stratum germinativum and the stratum corneum. These cells contained cytoplasmic FN particularly in the form of small granules. Our results support the view that MRC differ from other epidermal cells in their in vivo FN localizations.

Animals

Dermal tracts in frog skin: fibronectin pathways for cell migration.

The dermis of the frog skin (Rana esculenta) displayed a remarkable organization of vertical and horizontal tracts. Vertical thick tracts connected the dermal Stratum spongiosum with the subcutaneous tissue. Horizontal thin tracts were found alongside and contiguous to them. The thick tracts were sheathed by collagen fibrils of the Stratum compactum which were vertically oriented (i.e. parallel to the axes of the tracts) according to the horizontal and orthogonal arrangement of the collagen bundles of the Stratum compactum. The thin tracts devoid of collagenous sheath were formed by clear spaces between superimposed collagen bundles of the dermal Stratum compactum. On vertical sections, the thick tracts were seen to contain fibronectin (FN), detected by indirect immunoperoxidase. Continuous vertical FN lines were centred in these tracts. On horizontal sections, a clear zone around these FN-centred lines was also sheathed by FN. The thick tracts contained flattened pigmentary cells and fibroblasts; these cells were FN-outlined. The thin tracts contained patches of FN and FN-outlined fibroblasts. In culture, in vertical thick tracts, both pigmentary cells and fibroblasts disappeared when antiserum to FN was added to the culture medium. This suggested that thick tracts were pathways allowing pigmentary cells to move upward or downward between their usual upper dermal and lower subcutaneous localizations. Fewer fibroblasts were found in the thin tracts in the presence of antiserum to FN.

Animals

Fibronectin (FN) localizations in early wound healing of cultured frog skins.

The wound healing process of frog skin fragments in epibolic cultures has provided information on FN localizations during the migration of keratinocytes. Mainly two FN localizations were studied by indirect immunodetections: Epidermal localization around keratinocytes which have acquired a fibroblastic shape. Dermal localizations of the sectioned collagen of the stratum spongiosum and stratum compactum detected at the beginning of the culture. Both localizations were observed in this epibolic wound healing process during 6 hr and 24 hr in culture and showed a differential sensitivity to cycloheximide (CHX). It was worth noting that fibronectin was permanently detected in the subcutaneous tissue of non-cultured or cultured skin fragments with or without CHX.

Animals