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Biomedical subjects

C Zhao

Publications and source records attributed to C Zhao.

At least 181 records · Page 10Linked to original sources
PubMed1997

High-resolution imaging at the cellular and subcellular levels in flattened whole mounts of early zebrafish embryos.

We describe a rapid and sensitive method for high-resolution imaging at the cellular and subcellular levels in the whole-mount zebrafish embryo. The procedure involves fixing and staining the embryo, followed by deyolking and flattening it under a cover slip, to produce a planar mount that is 20 to 100 microns thick. Such a flattened whole mount allows imaging with a spatial resolution of approximately 500 nm in the x-y plane and does not require the use of embedding, sectioning, confocal microscopy, or computational deblurring procedures. We can resolve all individual nuclei and chromosome sets in the embryo, up to the late gastrula stage (10,000 cell stage). In addition, older embryos (through the segmentation stage) can also be examined, with the preservation of significant morphological detail. Because of its ability to resolve subcellular detail, the flattened whole-mount method can provide significant biological information beyond what can be obtained from conventional (three-dimensional) whole mounts. We have used the flattened whole-mount method to study subcellular events related to progression through the cell cycle or to apoptosis, in cells of the early zebrafish embryo. A specific DNA-binding dye (Hoechst 33258) or an antibody against a chromosomal protein (histone H1) was used to stain the nuclei of individual cells in the embryo. This allowed us to determine the spatial positions of all the individual cells, and also their stages in the cell cycle. A terminal transferase (TUNEL) assay was used to detect apoptotic cells. This combination of specific stains allowed us to study the behaviors of groups of cells in situ, within the developing zebrafish embryo.

Animals↗
PubMed1997

Changes in tachykinin--ergic nerve terminal densities in rat nasal mucosa during hypersensitivity.

OBJECTIVE: To further carry out a quantitative immunohistochemical study on neuropeptides (NPs) participation in perennial allergic rhinitis (PAR) in nasal mucosa. METHODS: By using light microscopy (routine hematoxylin and eosin staining, Wright's staining and 1% toluidine blue staining) and scanning as well as transmission electron microscopy, an exhaustive immunohistochemical and morphometric study was made on the changes of substance P (SP), neuropeptide A (NKA) and neuropeptide B (NKB) in the nasal mucosa of a toluene-2,4-iso-cyanate (TDI)-induced PAR model of rat. RESULTS: The results showed that the densities of all three peptidergic terminals were significantly increased (P < 0.01) in experimental group (115.72 +/- 28.43 for SP, 39.23 +/- 10.34 for NKA and 37.24 +/- 11.22 for NKB, respectively) as compared with those in control group (49.65 +/- 11.23 for SP, 12.44 +/- 6.21 for NKA and 13.31 +/- 7.60 for NKB, respectively). Increased staining, thickening of peptidergic terminals and enlargement of varicosities were found. Mast cell infiltration in nasal mucosa was in parallel with increase of peptidergic terminals. CONCLUSION: All results support the hypothesis that NP is a newly-recognized but potentially important factor in the pathogenesis of PAR.

Animals↗
PubMed1997

[Analysis of sensorineural hearing loss in 77 children].

The causes and cochlear-vestibular functions were analysed in 77 deaf children. The results showed that: the occurrence of hearing impairment beyond a moderately severe degree of deafness was 109 ears. The abnormalties of electronystagmus and posturography were 88.9% (44/45). 19 cases (38 ears) were large vestibular aquaduet syndromes, which were embryo development malformations. The average age of onset was 7.1 years. 15 cases have suffered from deafness of aminoglycoside ototoxicity. The average age of onset was 2.8 years. 13 of them appear to suffer deaf-mutism because of the early onset in their lives. The deaf mechanisms of the large vestibular aqueduct syndrome and the ototoxicity were discussed.

Adolescent↗
PubMed1996

Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells.

We compared the expression of human alpha- and beta-defensins by various human tissues. mRNA for alpha-defensins HNP1-3, abundant in bone marrow, was detected in peripheral blood leukocytes, spleen and thymus by RT-PCR, which revealed alpha-defensins HD5 and HD6 only in the small intestine. In contrast, the pancreas and kidney expressed high levels of hBD-1 and lower levels of this beta-defensin were found in many organs by RT-PCR (salivary gland > trachea > prostate and placenta > thymus, testis, small intestine). hBD-1 mRNA was produced constitutively by cultured normal human epithelial cells derived from the trachea, bronchi, small airways and the mammary gland. These largely non-overlapping tissue distributions of human alpha- and beta-defensins suggest that hBD-1 may be positioned to defend epithelial cells and mucosae from infection, whereas expression of HNP1-3 in neutrophils and HD5 and HD6 in Paneth cells allows these alpha-defensins to participate in systemic and small intestinal host defenses, respectively.

Amino Acid Sequence↗
PubMed1996

Novel azacyclic ureas that are potent inhibitors of HIV-1 protease.

A series of novel, azacyclic ureas which are highly potent inhibitors of the HIV-1 protease (IC50 = 4.1 to < 0.5 nM) were synthesized. Aqueous solubilities of this series of compounds were improved by incorporating polar functional groups at the P1' P2 and P2' positions. These compounds also possess good anti-viral activity by inhibition of the cytopathic effect of HIV-13B in MT-4 cells in vitro.

Amino Acid Sequence↗
PubMed1996

Growth-independent induction of spermidine transport by IL-4 and IL-13 in ZR-75-1 human breast cancer cells.

Polyamine transport is strongly induced by insulin and estradiol (E2) in ZR-75-1 human breast cancer cells. Because signal transduction mechanisms of insulin and interleukin-4 (IL-4) partly overlap, we have compared the ability of these agents as well as that of interleukin-13 (IL-13), a cytokine that often mimics IL-4, to modulate spermidine transport in these cells. In the presence of E2, insulin increased DNA content and the rate of [3H]spermidine uptake by 2.1- and 3.7-fold, respectively, after an 8-day incubation, whereas the sole addition of IL-4 caused a quantitatively similar induction of [3H]spermidine uptake while leaving cell growth unaffected. No comparable induction of spermidine transport was observed with interleukins-1 alpha and -6, and the effect of IL-4 was not additive to that elicited by insulin plus E2. IL-4 and IL-13 stimulated [3H]spermidine uptake to a comparable extent, with half-maximal effects observed at 80 and 400 pg/ml, respectively. Interferon-gamma inhibited IL-4- and IL-13-dependent spermidine uptake to a much greater extent than basal or insulin-induced transport of the polyamine. IL-4 and IL-13 increased the Vmax and K(m) of [3H]spermidine uptake by about 4- and 2.5-fold, respectively. Na(+)-dependent amino acid uptake was increased by insulin but not by IL-4 or IL-13, indicating that the cytokines do not induce a general increase in membrane transport activity. IL-4 and IL-13 did not interfere with feedback inhibition of polyamine uptake, and only modestly decreased polyamine content after prolonged incubation, suggesting that these cytokines stimulate spermidine uptake by increasing total transport capacity rather than by repressing and endogenous inhibitor.

Biological Transport↗
PubMed1996

Endotoxin and cytokines induce expression of leptin, the ob gene product, in hamsters.

The expression of leptin, the ob gene product, is increased in adipose tissue in response to feeding and energy repletion, while leptin decreases food intake. Because adipose tissue gene expression is regulated by cytokines induced during infection and because infection is associated with anorexia, we tested whether induction of leptin might occur during the host response to infection. Administration of endotoxin (LPS), a model for gram negative infections, induces profound anorexia and weight loss in hamsters. In fasted adipose tissue to levels similar to fed control animals. There is a strong inverse correlation between mRNA levels of leptin and subsequent food intake. TNF and IL-1, mediators of the host response to LPS, also induced anorexia and increased levels of leptin in mRNA in adipose tissue. As assessed by immuknoprecipitation and Western blotting, circulating leptin protein is regulated by LPS and cytokines in parallel to regulation of adipose tissue leptin mRNA. Induction of leptin during the host response to infection may contribute to the anorexia of infection.

Adipocytes↗
PubMed1996

A novel, picomolar inhibitor of human immunodeficiency virus type 1 protease.

The design, synthesis, and molecular modeling studies of a novel series of azacyclic ureas, which are inhibitors of human immunodeficiency virus type 1 (HIV-1) protease that incorporate different ligands for the S1', S2, and S2' substrate-binding sites of HIV-1 protease are described. The synthesis of this series is highly flexible in the sense that the P1', P2, and P2' residues of the inhibitors can be changed independently. Molecular modeling studies on the phenyl ring of the P2 and P2' ligand suggested incorporation of hydrogen-bonding donor/acceptor groups at the 3' and 4-positions of the phenyl ring should increase binding potency. This led to the discovery of compound 7f (A-98881), which possesses high potency in the HIV-1 protease inhibition assay and the in vitro MT-4 cell culture assay (Ki = approximately 5 pM and EC50 = 0.002 microM). This compares well with the symmetrical cyclic urea 1 pioneered at DuPont Merck.

Binding Sites↗
PubMed1996

Effect of prostatic growth factor, basic fibroblast growth factor, epidermal growth factor, and steroids on the proliferation of human fetal prostatic fibroblasts.

To study the relationship between androgen metabolism and the pathogenesis of benign prostatic hypertrophy, we purified a growth factor from benign hyperplastic tissue of human prostates and assayed the proliferative responses of human fetal prostatic fibroblasts to the purified growth factor (hPGF), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), dihydrotestosterone (DHT), and estradiol (E2). Prostatic tissue extracts were fractionated using heparin-Sepharose chromatography. The fraction that eluted with 1.3-1.7 M NaCl contained the majority of mitogenic activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE) of the lyophilyzed active fraction showed a band at 17,000 daltons. Human prostatic fibroblasts were isolated from fetal prostate and tested for their proliferative responses to hPGF, bFGF, EGF, DHT, and E2. hPGF, as well as bFGF and EGF, did increase tritiated thymidine incorporation into the cultured fibroblasts. DHT(10(-7) M) had a significant stimulatory effect on cell growth in serum-free media after 6 days of culture. E2(10-7 M) had no effect on cell proliferation. The combination of DHT and E2 showed no synergistic effect. We conclude that our purified hPGF, bFGF, and EGF promote cell growth directly, DHT indirectly, while E2 does not. The effect of DHT appears to be mediated via the increased production and/or secretion of growth factor(s). Possibly, the bFGF-like hPGF purified from human benign hyperplastic prostatic tissue is such a mediator.

Analysis of Variance↗
PubMed1996

Protective effects of calcium from nonfat dried milk against colon carcinogenesis in rats.

A number of studies have demonstrated the protective effect of dietary calcium against risk for colon cancer. The objective of this experimental study was to test the efficacy of two sources of dietary calcium, elemental calcium in the form of CaCO3 and dairy calcium as nonfat dried milk (NFDM), in colon tumor inhibition. Male weanling F344 rats were fed six test diets containing low (LF, 5%) and high (HF, 20%) levels of corn oil and low (0.5%) and high (1.0%) levels of calcium supplemented as CaCO3 or NFDM in a 2 x 3 factorial design. Tumors were induced with two weekly injections of azoxymethane at 12 mg/kg body wt. After 27 weeks on the test diets, animals were necropsied for tumor analysis. There was no difference in tumor incidence for fat or calcium source main effects, but a significant interaction was seen between fat and calcium source, with the lowest tumor incidence seen in the HF/NFDM group. Calcium compartmentalization studies demonstrated no effects of calcium on serum calcium levels but increased urinary and fecal water calcium in the higher-calcium diets. Increased dietary calcium also decreased fecal bile acid concentrations, but there was no effect on fecal water bile acids. Intermediate biomarkers of colon carcinogenesis were not affected by the dietary treatments except for fat effects on carcinogen-induced nuclear aberrations. These results indicate that source of calcium is not critical but that total dietary context may affect efficacy of calcium against colon carcinogenesis.

Animals↗
PubMed1996

Abnormally high platelet activity after discontinuation of acetylsalicylic acid treatment.

Production of 12-L-hydroxy-5, 8, 10-heptadecatrienoic acid (12-HHT) from platelets and bleeding times were studied in 32 males during acetylsalicylic acid (ASA) treatment and 1 and 2 weeks after withdrawal. All patients (age 42-77 years) had ASA treatment because of angina pectoris. The metabolite 12-HHT is formed in the same amount as the proaggregatory and vasoactive metabolite thromboxane A2. Initially the daily ASA dose was 75 mg (n = 15), 160 mg (n = 12) or 250-300 mg (n = 5). In all patients, median 12-HHT level increased from 40 to 240g/750 x 10(6) platelets (P < 0.001) 1 week after withdrawal of ASA, and four patients had abnormally high values. Median bleeding time decreased from 312 to 268 s (P = 0.003) in the 75 mg group and from 315 to 235 s in the 160 mg group (P = 0.01). Two weeks after withdrawal of ASA, median 12-HHT was 390g/750 x 10(6) platelets and eight patients (25%) had abnormally high values. One patient still had a prolonged bleeding time. Wide interindividual variations were observed in all groups. Our results indicate that rapid withdrawal of ASA, may cause abnormally high 12-HHT levels reflecting increase of thromboxane A2 with possible hazardous effects in patients with cardiovascular disease.

Adult↗
PubMed1996

Inhibitory activity of the equine infectious anemia virus major 5' splice site in the absence of Rev.

The major 5' splice site of equine infectious anemia virus (EIAV) conforms to the consensus 5' splice site in eight consecutive positions and is located immediately upstream of the gag AUG. Our results show that the presence of this 5' splice site on the EIAV gag mRNA decreases Gag production 30- to 60-fold. This is caused by inefficient nuclear mRNA export and inefficient mRNA utilization. Inhibition could be overcome by providing human immunodeficiency virus type 1 Rev/Rev-responsive element, human T-cell leukemia virus type 1 Rex/Rex-responsive element, or simian retrovirus type 1 constitutive transport element. In addition, inhibition could be abolished by introducing single point mutations in the 5' splice site or by moving the 5' splice site away from its natural position immediately upstream of the gag AUG. This demonstrates that both maintenance of a perfect consensus 5' splice site and its proper location on the mRNA are important for inhibitory activity of the EIAV major 5' splice site.

Base Sequence↗
PubMed1996

Identification of nuclear and cytoplasmic proteins that interact specifically with an AU-rich, cis-acting inhibitory sequence in the 3' untranslated region of human papillomavirus type 1 late mRNAs.

Expression of human papillomavirus late genes encoding L1 and L2 capsid proteins is restricted to terminally differentiated epithelial cells. We have previously identified and characterized an AU-rich, cis-acting negative regulatory element in the 3' untranslated region of human papillomavirus type 1 late mRNAs. This element acts posttranscriptionally to reduce mRNA levels and the translation efficiency of mRNAs. The experiments reported here are a continuation of our previous work. We have used RNA gel shifts and UV cross-linking assays to identify cellular proteins that interact with the inhibitory RNA sequence of human papillomavirus type 1. RNA gel shift assays established that cellular proteins interact with the AU-rich sequence. The binding of nuclear proteins was inhibited by competition with poly(U), whereas the binding of cytoplasmic proteins was inhibited by competition with poly(U) and also by competition with poly(A) and poly(G). Two nuclear proteins and two cytoplasmic proteins that bind specifically to the AU-rich RNA sequence were identified by UV cross-linking. These proteins did not bind to the 3' untranslated region of human papillomavirus type 1 early mRNAs, which does not show inhibitory activity. The cellular proteins identified in our experiments may therefore be involved in the inhibition of human papillomavirus type 1 late gene expression in nondifferentiated epithelial cells.

Base Sequence↗
PubMed1996

Erythropoietin-like activity in vivo of the fusion protein rhIL-6/IL-2 (CH925).

CH925 is a novel cytokine of a fusion protein interleukin-6 (IL-6)/IL-2 exhibiting erythropoietin (Epo)-like effects in vivo and ex vivo, in addition to its enhanced effects compared to IL-2 and IL-6 reported by us previously, which indicates its potential clinical use. Our present study was undertaken to determine the Epo-like activity of CH925 in vivo. The reticulocyte response was observed in transfusion-induced polycythemic mice by using flow cytometry with pyronin Y staining. On day 2 after injection of CH925, the average number of reticulocytes was 2.11% in the group given 250 micrograms/kg/d and 1.01% for 100 micrograms/kg/d. The mean fluorescence intensity (MFI) also significantly increased. Longitudinal studies of CH925 were performed on days 2, 4, and 10, and reticulocyte counts increased up to a peak on day 4. Activity of CH925 (100 micrograms/kg/d) corresponds to 1 U of standard rhEpo in our study.

Animals↗
PubMed1996

[Effects of ACM sequential chemotherapy on ovarian function in trophoblastic tumors].

OBJECTIVE: To study the changes of ovarian function during and after ACM sequential chemotherapy on trophoblastic tumors. METHODS: 17 patients with trophoblastic tumors, received totally 48 cycles of ACM chemotherapy. Of these, 7 underwent hysterotomy and focal ectomy and ovarian biopsy. Changes of menstruation and BBT were observed. beta-hCG, E2, FSH and LH were assayed and histological and immunohistochemical studies in ovaries biopsied were made. RESULTS: Amenorrhea and anovulatory BBT were predominant at the onset of chemotherapy and high level of beta-hCG. Following chemotherapy and decrease of beta-hCG, menstruation recovered with less amount and BBT transformed into ovulatory types with shorter luteal phase. The E2 level appeared to be lower and the FSH and LH level higher. The different kinds of follicles were decreased, but estrogen receptors unchanged. Follow-up showed that ovarian function of 12 of the 17 patients returned to normal in one year. CONCLUSION: ACM chemotherapy suppresses ovarian function mildly and temporarily. Trophoblastic tumor also affects ovarian function. ACM chemotherapy seems to be suitable for young patients with trophoblastic tumors who desire child-bearing.

Adult↗