[The new progress in the study of calcitonin gene-related peptide].
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Adjuvants, Immunologic↗
Biomedical subjects
M Zhao
Publications and source records attributed to M Zhao.
[Study on activity of inner inhibitory substances of Acanthopanax senticosus (Rupr. et Maxim) harms fruits and seeds].
OBJECTIVE: By studying on the inner inhibitory substances of Acanthopanax senticosus fruits and seeds, the reason for dormancy of the Seeds was investigated. METHOD: First, the raw extracts of Acanthopanax senticosus fruits and seeds were prepared and determined by biological methods. Then, the ether extract of Acanthopanax senticosus fruits was prepared and the components were separated by paper chromatography and each section with different Rf value was determined, respectively. RESULT AND CONCLUSION: Inner inhibitory substances with high activity exists in Acanthopanax senticosus fruits and their outer coats, middle coats and the seeds. The inner inhibitory substances showed stronger inhibitory effect on the germination of Brassica chinensis seeds than the growth of its younger roots. The Rf 0.1 section of the ether extract of Acanthopanax senticosus fruit coats showed the strongest inhibitory activity on the germination of Brassica chinensis seeds and the growth of its young roots. The rough extracts of Acanthopanax senticosus fruits could inhibit the activity of the (alpha-amylase in the process of wheat seeds germination. The dormanvy of Acanthopanax senticosus seeds is related to the inhibitory substances with high activity in Acanthopanax senticosus fruits and seeds.
Brassica↗
Urotensin II receptor in the rat airway smooth muscle and its effect on the rat airway smooth muscle cells proliferation.
OBJECTIVE: To investigate the characteristics of urotensin II (U-II) receptor in the rat airway smooth muscle and the effect and signal transduction pathway of U-II on the proliferation of airway smooth muscle cells. METHODS: Using 125I-UII binding assay to measure the Bmax and Kd of U-II receptor. Using the 3H-TdR incorporation to determine the effect of U-II on the proliferation of airway smooth muscle cells and its signal transduction pathway. Using Fura-2/AM to measure the effect of U-1I on the cytosolic free calcium concentration. RESULTS: 1. 125I-UII binding increased with the time and reached saturation at 45 min. The B(max0 was (11.36 +/- 0.37)fmol/mg pr and Kd was (4.46 +/- 0.61) nmol/L. 2. U-II increased 3H-TdR incorporation of the airway smooth muscle cells in a dose-dependent manner. 3. H7, PD98059 and nicardipine, inhibitors of PKC, MAPK, calcium channel, respectively, significantly inhibited U-II-stimulated 3H-TdR incorporation of airway smooth muscle cells. W7, inhibitor of CaM-PK, had no effect. 4. Cyclosporin A, inhibitor of CaN, inhibited 3H-TdR incorporation of the airway smooth muscle cells induced by U-II in a dose-dependent manner. 5. U-II promoted cytosolic free calcium concentration increase by 18%. CONCLUSIONS: 1. There was U-II receptor in the rat airway smooth muscle. 2. The effect of U-II-stimulated 3H-TdR incorporation of airway smooth muscle cells was mediated by such signal transduction pathway as Ca2+, PKC, MAPK and CaN, etc.
Animals↗
[Impact of AFP promoter on the expressions of GFP in tumor cells].
OBJECTIVE: To study the impact of AFP 5'flanking promoter (enhancer) on the expression of GFP in hepatocarcinoma cell. METHODS: Green Fluorescent Protein (GFP) reporter gene expression plasmid pcDNA3-GFP-AFP-w under the direction of AFP 5' flanking promoter (enhancer) was constructed by recombinant DNA technology and confirmed by restriction analyses. pcDNA3-GFP-AFP-w, pcDNA3-GFP and pcDNA3 were transfected into Hela and Bel7402 cells by lipofectin and selected by G418 respectively, after amplification of the positive cell clones, expression of GFP was detected by Western blotting and quantitatively analysed by GEL Doc 2000 digital image systems. RESULTS: The expression of GFP was lower in Bel-GFP-AFP-w than in Bel-GFP but was significantly higher than in Hela-GFP-AFP-w. CONCLUSION: GFP reporter gene plasmid pcDNA3-GFP-AFP-w under the direction of the 3.1 kb AFP 5'flanking promoter (enhancer) can be expressed in HCC Bel7402 cell definitely and specifically.
Carcinoma, Hepatocellular↗
Protection against oxidant-mediated lysosomal rupture: a new anti-apoptotic activity of Bcl-2?
Bcl-2 antagonizes apoptosis through mechanisms which are not completely understood. We have proposed that apoptosis is initiated by minor lysosomal destabilization followed some time later by secondary massive lysosomal rupture. In J774 cells over-expressing Bcl-2, early oxidant-induced lysosomal destabilization is unaffected but secondary lysosomal rupture and apoptosis are suppressed, despite the fact that wild-type and Bcl-2 over-expressing cells degrade hydrogen peroxide at similar rates. It may be that Bcl-2 directly blocks the effects of released lysosomal enzymes and/or prevents downstream activation of unknown cytosolic pro-enzymes by released lysosomal hydrolases, suggesting a new and heretofore unknown activity of Bcl-2.
Acridine Orange↗
Binding of spin-labeled galactosides to the lactose permease of Escherichia coli.
A series of nitroxide spin-labeled alpha- or beta-galactopyranosides and a nitroxide spin-labeled beta-glucopyranoside have been synthesized and examined for binding to the lactose permease of Escherichia coli. Out of the twelve nitroxide spin-labeled galactopyranosides synthesized, 1-oxyl-2, 5, 5-trimethyl-2-[3-nitro-4-N-(hexyl-1-thio-beta-D-galactopyranosid-1 -yl )]aminophenyl pyrrolidine (NN) exhibits the highest affinity for the permease based on the following observations: (a) the analogue inhibits lactose transport with a K(I) about 7 microM; (b) NN blocks labeling of single-Cys148 permease with 2-(4'-maleimidylanilino) naphthalene-6-sulfonic acid (MIANS) with an apparent affinity of about 12 microM; (c) electron paramagnetic resonance demonstrates binding of the spin-labeled sugar by purified wild-type permease in a manner that is reversed by nonspin-labeled ligand. The equilibrium dissociation constant (K(D)) is about 23 microM and binding stoichiometry is approximately unity. In contrast, the nitroxide spin-labeled glucopyranoside does not inhibit active lactose transport or labeling of single-Cys148 permease with MIANS. It is concluded that NN binds specifically to lac permease with an affinity in the low micromolar range. Furthermore, affinity of the permease for the spin-labeled galactopyranosides is directly related to the length, hydrophobicity, and geometry of the linker between the galactoside and the nitroxide spin-label.
Alkylating Agents↗
Absence of cardiolipin in the crd1 null mutant results in decreased mitochondrial membrane potential and reduced mitochondrial function.
Cardiolipin (CL) is a unique phospholipid which is present throughout the eukaryotic kingdom and is localized in mitochondrial membranes. Saccharomyces cerevisiae cells containing a disruption of CRD1, the structural gene encoding CL synthase, have no CL in mitochondrial membranes. To elucidate the physiological role of CL, we compared mitochondrial functions in the crd1Delta mutant and isogenic wild type. The crd1Delta mutant loses viability at elevated temperature, and prolonged culture at 37 degrees C leads to loss of the mitochondrial genome. Mutant membranes have increased phosphatidylglycerol (PG) when grown in a nonfermentable carbon source but have almost no detectable PG in medium containing glucose. In glucose-grown cells, maximum respiratory rate, ATPase and cytochrome oxidase activities, and protein import are deficient in the mutant. The ADP/ATP carrier is defective even during growth in a nonfermentable carbon source. The mitochondrial membrane potential is decreased in mutant cells. The decrease is more pronounced in glucose-grown cells, which lack PG, but is also apparent in membranes containing PG (i.e. in nonfermentable carbon sources). We propose that CL is required for maintaining the mitochondrial membrane potential and that reduced membrane potential in the absence of CL leads to defects in protein import and other mitochondrial functions.
Cardiolipins↗
Big mitogen-activated kinase regulates multiple members of the MEF2 protein family.
Big mitogen-activated protein (MAP) kinase (BMK1), a member of the mammalian MAP kinase family, is activated by growth factors. The activation of BMK1 is required for growth factor-induced cell proliferation and cell cycle progression. We have previously shown that BMK1 regulates c-jun gene expression through direct phosphorylation and activation of transcription factor MEF2C. MEF2C belongs to the myocyte enhancer factor 2 (MEF2) protein family, a four-membered family of transcription factors denoted MEF2A, -2B, -2C, and -2D. Here, we demonstrate that, in addition to MEF2C, BMK1 phosphorylates and activates MEF2A and MEF2D but not MEF2B. The blocking of BMK1 signaling inhibits the epidermal growth factor-dependent activation of these three MEF2 transcription factors. The sites phosphorylated by activated BMK1 were mapped to Ser-355, Thr-312, and Thr-319 of MEF2A and Ser-179 of MEF2D both in vitro and in vivo. Site-directed mutagenesis reveals that the phosphorylation of these sites in MEF2A and MEF2D are necessary for the induction of MEF2A and 2D transactivating activity by either BMK1 or by epidermal growth factor. Taken together, these data demonstrate that, upon growth factor induction, BMK1 directly phosphorylates and activates three members of the MEF2 family of transcription factors thereby inducing MEF2-dependent gene expression.
Amino Acid Sequence↗
Methioninase gene therapy of human cancer cells is synergistic with recombinant methioninase treatment.
Results obtained over the past 40 years have demonstrated that tumor cells of all types tested have an elevated growth requirement for methioninase compared with normal cells. Recombinant methioninase (rMETase) cloned from Pseudomonas putida has been found previously to be an effective antitumor agent attributable to deprivation of the extracellular methionine source of the tumor. To degrade intracellular methioninase, we have now developed an adenoviral vector inserted with the P. putida methioninase (MET) gene (rAd-MET). The in vitro efficacy of rAd-MET was tested on the OVCAR-8 human ovarian cancer cell line, the HT1080 human fibrosarcoma cell line, and human normal fibroblasts. rAd-MET transduction of OVACAR-8 and HT1080 resulted in high levels of methioninase expression up to 10% or more of the total protein of the cells, depending on the multiplicity of infection. The IC50 of rAd-MET for OVCAR-8 cells in 96-well plates was approximately 2 x 106 plaque-forming units (pfu)/well. The IC50 of control adenovirus (control-rAd) was 4 x 10(7) pfu/well, 20 times higher than rAd-MET. In the presence of the IC50 of 2 x 10(6) pfu/well of rAd-MET, the addition of 0.025 units/ml of rMETase, which is 25% of the IC50, resulted in a 90% inhibition of tumor cell number. This indicated that rAd-MET enhanced the efficacy of rMETase. In contrast, 2 x 10(6) pfu/well of control-rAd in combination with 0.025 units/ml of rMETase had an efficacy of only 10% inhibition of cell number. The synergistic effect of the combination of rMETase and rAd-MET was quantitated by calculating the combination index (CI). The CIs for all combinations of rAd-MET and rMETase tested on OVCAR-8 were <0.7 with a mean of 0.5, indicating synergy. Similar synergy of rAd-MET and rMETase was seen on HT1080 human fibrosarcoma cells with a mean of 0.74. In contrast, the CIs of all combinations of rMETase and control adenovirus concentrations tested on both cell lines had a mean CI of approximately 1, which indicated that this combination had only an additive effect. The normal fibroblasts, on the other hand, appeared relatively resistant to the MET gene because in the presence of rMETase, 2.5 x 10(7) pfu/well of rAd-MET or control rAd had almost an identical effect on cell survival. The selectively strong synergy of rAd-MET and rMETase on cancer cells allows reduced levels of each agent to be used, thus decreasing potential side effects.
Carbon-Sulfur Lyases↗
Reduced expression of p27(Kip1) correlates with an early stage of cancer invasion in oral squamous cell carcinoma.
Down-regulation of p27(Kip1) has been reported to correlate with poor survival of various carcinoma patients including oral squamous cell carcinomas (OSCCs). It is still unclear, however, at what stage of oral carcinogenesis the down-regulation of this protein occurs. In this study, therefore, we evaluated immunoexpression of p27(Kip1) protein in 17 cases of oral epithelial dysplasia and succeeding invasive OSCC in the same patient. We reported here that 88% cases showed high p27(Kip1) expression in dysplastic lesions, whereas 82% cases of succeeding invasive OSCC exhibited reduced expression. The reduction of p27(Kip1) expression was also observed in 16 of 19 (84%) early invasive lesions and well correlated with Ki-67 expression which is good indicator of cell proliferation. We also investigated immunoexpression of p53 protein of which abnormality has been known to occur during the early stage of OSCC development. Overexpression of p53 protein was demonstrated in 29% of dysplastic lesions, 42% of early invasive and 71% of invasive OSCCs. These findings suggest that abnormalities of both p53 and p27(Kip1) are involved in the carcinogenesis of OSCC, but they seem to play their role at different stages of oral cancer development, respectively. Reduced expression of p27(Kip1) may concern the cancer invasion directly or indirectly as well as abnormal proliferation.
Carcinoma, Squamous Cell↗
Diabetes-associated mutations in a beta-cell transcription factor destabilize an antiparallel "mini-zipper" in a dimerization interface.
Maturity-onset diabetes of the young, a monogenic form of Type II diabetes mellitus, is most commonly caused by mutations in hepatic nuclear factor 1alpha (HNF-1alpha). Here, the dimerization motif of HNF-1alpha is shown to form an intermolecular four-helix bundle. One face contains an antiparallel coiled coil whereas the other contains splayed alpha-helices. The "mini-zipper" is complementary in structure and symmetry to the top surface of a transcriptional coactivator (dimerization cofactor of homeodomains). The bundle is destabilized by a subset of mutations associated with maturity-onset diabetes of the young. Impaired dimerization of a beta-cell transcription factor thus provides a molecular mechanism of metabolic deregulation in diabetes mellitus.
Amino Acid Sequence↗
Re-orientation and faster, directed migration of lens epithelial cells in a physiological electric field.
The vertebrate lens drives current through itself in a pattern which concentrates current efflux at the lens equator. Lens epithelial cells (LECs) move into this region where they change shape and differentiate into lens fibre cells. The mechanisms underpinning these cell behaviors are unclear. We have attempted to mimic, in isolation, the effects which such electrical signals have on LEC behaviors, by culturing LECs in a physiological DC electric field (EF) similar to that in lens. Primary human (PHLECs), primary bovine (PBLECs) and a transformed human cell line (THLECs) all changed shape to lie perpendicular to the EF, the same orientation which LECs adopt with respect to the equatorial EF as they differentiate into lens fibre cells. Exposure to an EF also significantly increased the migration rate of all three LEC types. All three LECs also showed directed cell migration although, curiously, different cell types moved in different directions. PBLECs and THLECs showed voltage-dependent, anode-directed migration, with a response threshold between 100-150 mV mm(-1)and 25-50 mV mm(-1), respectively. Small sheets of THLECs also migrated anodally. By contrast PHLECs migrated cathodally with a response threshold below 100 mV mm(-1). Reversing the polarity reversed the migration direction for each cell type. These observations raise three possibilities: (1) that small electric field may be one of the cues regulating lens epithelial cell behaviors in vivo; (2) that altering the in vivo electric field by lens replacement may contribute to the aberrant migration of epithelial cells in conditions such as posterior capsule opacification and (3) that applying electric fields may be one way of controlling aberrant lens epithelial cell behaviors.
Animals↗
Immunoexpression of transforming growth factor beta in desmoplastic ameloblastoma.
Desmoplastic ameloblastoma (DA) is an unusual subtype of ameloblastoma histologically characterized by the pronounced collagenized stroma. In the present study, the immunolocalization of transforming growth factor beta (TGF-beta), one of the most potent local factors for modulating extracellular matrix formation, was observed in DA in order to study its participation in the stromal desmoplasia. Seven cases of DA, including a "hybrid" lesion, were studied together with ten cases of ordinary follicular and plexiform ameloblastomas as the control. In contrast to ordinary ameloblastomas, marked immunoexpression was observed in all DAs but one. In the "hybrid" lesion, TGF-beta was not expressed in the area of follicular ameloblastoma but in that of DA. These results show that TGF-beta produced by tumor cells of DA plays a part in the desmoplastic matrix formation.
Adolescent↗
Immunohistochemical demonstration of an enamel sheath protein, sheathlin, in odontogenic tumors.
Enamel proteins can be useful markers for assessment of the functional differentiation of neoplastic epithelium and the nature of extracellular matrices in odontogenic tumors. In the present study, we examined immunohistochemical localization of sheathlin, a recently cloned enamel sheath protein, in various odontogenic tumors to evaluate functional differentiation of tumor cells and the nature of hyalinous or calcified matrices in odontogenic neoplasms. Distinct immunolocalization of sheathlin was observed in the immature enamel of the tooth germ at the late bell stage. Secretory ameloblasts facing the enamel matrix also showed positive staining in their cytoplasm. Definite localization of sheathlin was demonstrated in the enamel matrix in odontogenic tumors with inductive dental hard tissue formation such as ameloblastic fibroodontomas and odontomas. Immunoexpression of sheathlin was, furthermore, demonstrated in eosinophilic droplets in solid nests of adenomatoid odontogenic tumor (AOT) and ghost cells in the epithelial lining of calcifying odontogenic cyst (COC). In AOT, cells facing the eosinophilic droplets also expressed the protein in their cytoplasm. There was neither intracellular staining for sheathlin in the tumor cells nor extracellular staining in the matrix of ameloblastomas and calcifying epithelial odontogenic tumors. Dentin, dysplastic dentin-like hyaline material and cementum in the tumors examined were negative for sheathlin. These results show that immunodetection of sheathlin is a useful marker for functional differentiation of secretory ameloblasts and enamel matrix, which is often hard to differentiate from other hard tissues in odontogenic tumors. Our findings from the view point of sheathlin expression support that the tumor cells of ameloblastomas do not attain full differentiation into functional ameloblasts. It is very interesting that epithelial cells in odontogenic tumors can differentiate into functional ameloblasts without induction by odontogenic mesenchyme, as shown by immunoexpression of sheathlin in eosinophilic droplets within solid epithelial sheets in AOT and ghost cells in the epithelial lining of COC where inductive participation of mesenchymal cells was most unlikely.
Ameloblastoma↗
Hemispheric asymmetries and individual differences in visual concept learning as measured by functional MRI.
Dynamic changes in brain regions active while learning novel visual concepts were examined in humans using functional magnetic resonance imaging. Participants learned to distinguish between exemplars of two categories, formed as distortions of different unseen prototype stimuli. Regions of the right hemisphere (dorsolateral prefrontal and inferior parietal areas) were active early in learning and throughout task performance, whereas homologous portions of the left hemisphere were active only in later stages of learning. Left dorsolateral prefrontal activation was found only in participants who showed superior conceptual learning. Such a progression from initial right-hemisphere processing of specific instances to bilateral activity as left-hemisphere conceptual processes are recruited may underlie the development of many forms of visual knowledge.
Adult↗
Improved phase-contrast flow quantification by three-dimensional vessel localization.
In this paper, a method of three-dimensional (3D) vessel localization is presented to allow the identification of a vessel of interest, the selection of a vessel segment, and the determination of a slice orientation to improve the accuracy of phase-contrast magnetic resonance (PCMR) angiography. A marching-cube surface-rendering algorithm was used to reconstruct the 3D vasculature. Surface-rendering was obtained using an iso-surface value determined from a maximum intensity projection (MIP) image. This 3D vasculature was used to find a vessel of interest, select a vessel segment, and to determine the slice orientation perpendicular to the vessel axis. Volumetric flow rate (VFR) was obtained in a phantom model and in vivo using 3D localization with double oblique cine PCMR scanning. PCMR flow measurements in the phantom showed 5. 2% maximum error and a standard deviation of 9 mL/min during steady flow, 7.9% maximum error and a standard deviation of 13 mL/min during pulsatile flow compared with measurements using an ultrasonic transit-time flowmeter. PCMR VFR measurement error increased with misalignment at 10, 20, and 30 degrees oblique to the perpendicular slice in vitro and in vivo. The 3D localization technique allowed precise localization of the vessel of interest and optimal placement of the slice orientation for minimum error in flow measurements.
Blood Flow Velocity↗
Reduced expression of CD44 variant 9 is related to lymph node metastasis and poor survival in squamous cell carcinoma of tongue.
Expression of CD44v9 was immunohistochemically studied in 120 biopsy specimens from primary squamous cell carcinoma (SCC) of the tongue and correlated with clinicopathological findings of the SCCs. The tumors were classified into three groups according to immunostaining pattern of CD44v9; 53 cases with distinct positivity in all cancer cells except for those in the central part of nests (Group 1, non-reduced group), 42 cases with reduced expression in peripheral cells of nests (Group 2, reduced group), and 25 cases with complete disappearance of the expression in one or more nests (Group 3, negative group). Nineteen of 25 (76%) tumors in Group 3 and 14 of 42 (33%) in Group 2 exhibited lymph node metastasis, compared with only 8 of 53 (15%) in Group 1. The average survival time in Groups 1, 2 and 3 was 4496+/-204, 3866+/-379 and 2719+/-359 days, respectively and became shorter with the reduction of CD44v9 expression. These results suggest that the down-regulation of CD44v9 in SCC of the tongue may relate to the detachment of tumor cells from primary lesions, establishment of lymph node metastasis and consequently the death of patients.
Adult↗
Ghost cells in calcifying odontogenic cyst express enamel-related proteins.
The so-called ghost cell is a unique cell type occurring in a variety of odontogenic and non-odontogenic lesions. However, the true nature of ghost cells has not been determined. In the present study, we examined the immunoreactivity of ghost cells in calcifying odontogenic cysts and dermal calcifying epitheliomas, with antibodies against amelogenin, enamelin, sheath protein (sheathlin) and enamelysin, in an attempt to clarify the nature of this unique cell. The cytoplasm of ghost cells in calcifying odontogenic cysts demonstrated distinct immunolocalization of the enamel-related proteins, while similar in the calcifying epitheliomas of the skin showed a negative reaction. The results indicate that the ghost cells in calcifying odontogenic cysts, as opposed to ghost cells in dermal calcifying epitheliomas, contain enamel-related proteins in their cytoplasm accumulated during the process of pathological transformation.
Amelogenin↗