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H Zhao

Publications and source records attributed to H Zhao.

At least 523 records · Page 29Linked to original sources
PubMed1994

NADPH-diaphorase localization in the olfactory system.

NADPH-diaphorase staining was studied in the intact olfactory system and following transections of the olfactory nerve. Intense staining was found in the olfactory epithelia of rat and salamander, especially at the mucosal surface. The olfactory bulb nerve and glomerular layers were also stained. Transection of the olfactory nerve in salamander, or bulbectomy in rat, resulted in decreased staining at the mucosal surface, although staining increased in cells deeper in the epithelium. Deafferentation also caused the disappearance of staining in the olfactory nerve and glomerular layers of the bulb. These results support the notion of a role for NADPH in the olfactory system, perhaps as a biosynthesis substrate for NO modulation of cGMP in the developing epithelium and a complementary role for CO in the adult.

Animals↗
PubMed1994

A tyrosine-based motif in the cytoplasmic domain of the alphavirus envelope protein is essential for budding.

The budding of enveloped viruses from cellular membranes is believed to be dependent on the specific interaction between transmembrane spike proteins and cytoplasmic core components of the virus. We found that the cytoplasmic domain of the E2 transmembrane spike glycoprotein of Semliki Forest virus contains two essential determinants which are absolutely needed for budding. The first constitutes a single tyrosine residue in the context of a direct pentapeptide repeat. The tyrosine could only partially be substituted for other residues with aromatic or bulky hydrophobic side chains, although these immediately reverted to the original genotype. The second determinant involves palmitylated cysteine residues flanking the tyrosine repeat motif. The function of these is probably to anchor the tail against the inner surface of the membrane so that the tyrosine-containing motif is properly presented to the nucleocapsid. This is the first example where a membrane virus employs a tyrosine signal for the selective incorporation of spike proteins into budding structures.

Amino Acid Sequence↗
PubMed1994

Oligomerization of an avian thymic parvalbumin. Chemical evidence for a Ca(2+)-specific conformation.

CPV3, the third parvalbumin isoform to be identified in the chicken, is produced exclusively in the thymus gland. Although parvalbumins are typically cysteine-deficient, CPV3 contains two cysteine residues, at positions 18 and 72. The reported three-dimensional parvalbumin structures suggest that the side chain of cysteine-72 should be solvent-accessible. Accordingly, we find that CPV3 readily forms disulfide-linked oligomers in the absence of reducing agents. The reaction, employing either O2 or ferricyanide ion as the oxidant, is apparently restricted to the Ca(2+)-bound form of the protein. The differing reactivity of the Ca2+, Mg2+, and apo-forms has significant structural implications.

Animals↗
PubMed1994

The activity of parafusin is distinct from that of phosphoglucomutase in the unicellular eukaryote Paramecium.

In this paper we identified the presence of a Paramecium phosphoglucomutase enzymatic activity which is clearly distinct from that of parafusin-the exocytosis-related phosphoglycoprotein. Since the recently cloned parafusin showed homology to rabbit muscle phosphoglucomutase, we have designed a specific peptide parafusin antibody-generated to a region not present in any known sequenced phosphoglucomutases-to distinguish parafusin from the Paramecium phosphoglucomutase. Separation of these two proteins was obtained using liquid chromatography, enzymatic activity assay and immunoblotting analysis with the specific parafusin peptide antibody. Parafusin fractions incorporated [B35S]UDP-Glc but not [35S]Glc-1-P whereas Paramecium phosphoglucomutase fractions incorporated [35S]Glc-1-P but not [B35S]UDP-Glc. This indicates that these two proteins are separate entities exhibiting different properties and most likely distinct functions in the cell.

Animals↗
PubMed1994

Novel avian thymic parvalbumin displays high degree of sequence homology to oncomodulin.

CPV3 is a novel avian parvalbumin. It displays an isoelectric point of 4.6, intermediate between that of avian thymic hormone (pI = 4.3) and the muscle parvalbumin isoform (pI = 5.2). Expression of CPV3, like that of avian thymic hormone (ATH), is restricted to the thymic stroma. However, the CPV3 content of chicken thymus tissue (120 micrograms/g tissue) is 4 times lower than that of ATH (500 micrograms/g tissue). The polymerase chain reaction (PCR) was used to gain access to the nucleotide sequence of CPV3. A 147-base pair fragment of the coding sequence, corresponding to residues 48-97, was amplified from total chicken cDNA using degenerate PCR primers. A RACE-PCR strategy was then used to extend the known sequence in both the 5' and 3' directions. The cDNA sequence thus obtained includes 671 base pairs. Primer extension analysis suggests that the cloned cDNA corresponds to a full-length transcript. Northern analysis of chicken mRNA indicates that the average CPV3 transcript is approximately 800 nucleotides in length, significantly smaller than the ATH message (approximately 1000 nucleotides). Southern analysis suggests the presence of a single CPV3 gene in the chicken genome. The translated nucleotide sequence, displaying 108 residues between the initiator and termination codons, is that of a beta-parvalbumin. The CPV3 sequence exhibits 58% identity with ATH and 52% identity with the chicken muscle isoform. Interestingly, CPV3 and the mammalian oncodevelopmental parvalbumin called oncomodulin are identical at 73 of 108 residues (68% identity). Correspondingly, flow-dialysis measurements with 45Ca2+ indicate that the Ca(2+)-binding domains are inequivalent, as in oncomodulin. The apparent dissociation constants, at pH 7.4 in 150 mM NaCl, are approximately 10 nM and 80 nM.

Amino Acid Sequence↗
PubMed1994

Assembly and entry mechanisms of Semliki Forest virus.

The alphavirus Semliki Forest (SFV) is an enveloped virus with a positive single-stranded RNA genome. The genome is complexed with 240 copies of a capsid protein into a nucleocapsid structure. In the membrane the virus carries an equal number of copies of a membrane protein heterodimer. The latter oligomers are grouped into clusters of three. These structures form the spikes of the virus and carry its entry functions, that is receptor binding and membrane fusion activity. The membrane protein heterodimer is synthesized as a p62E1 precursor protein which upon transport to the cell surface is cleaved into the mature E2E1 form. Recent studies have given much new information on the assembly and entry mechanism of this simple RNA virus. Much of this work has been possible through the construction of a complete cDNA clone of the SFV genome which can be used for in vitro transcription of infectious RNA. One important finding has been to show that a spike deletion variant and a capsid protein deletion variant are budding-negative when expressed separately but can easily complement each other when transfected into the same cell. This shows clearly that enveloped viruses use different budding strategies: one which depends on a nucleocapsid-spike interaction as exemplified by SFV and another one which is based on a direct core-lipid bilayer interaction as shown before to be the case with retroviruses. Another important finding concerns the activation process of the presumed fusion protein of SFV, the E1 subunit. In the original p62E1 heterodimer E1 is completely inactive. Activation proceeds in several steps. First p62 cleavage activates the potential for low pH inducible fusion. Next the low pH which surrounds incoming virus in endosomes induces dissociation of the heterodimeric structure. This is followed by a rearrangement of E1 subunits into homotrimers which are fusion active.

Animals↗
PubMed1994

Plasma renin activity and insulin sensitivity in normotensive subjects.

Insulin sensitivity in essential hypertension and atherosclerosis has elicited substantial research interest. Hyperinsulinemia in prospective studies identifies patients at risk for the development of coronary atherosclerosis. Plasma renin activity has also been shown to predict myocardial infarction, and correlates inversely with insulin sensitivity in hypertensive patients. The current study investigated the relationship between plasma renin activity and insulin sensitivity in 25 normotensive subjects. Using the euglycemic clamp technique, the data show a significant inverse correlation between supine plasma renin activity and insulin-mediated glucose uptake (r = -0.44; P = .027). Significant relationships were also found between plasma renin activity and fasting insulin concentration and fasting serum triglycerides. Reductions in relative skeletal muscle blood flow during insulin infusion have been reported in insulin-resistant compared to insulin-sensitive subjects. Skeletal muscle is the main site of insulin action during euglycemic clamp study and angiotensin II increases skeletal muscle blood flow. Increasing plasma renin activity may reflect a metabolic compensation which offsets the relative reduction in insulin-mediated enhancement in skeletal muscle blood flow in insulin resistance by generation of angiotensin II, which in turn promotes skeletal muscle blood flow.

Adult↗
PubMed1994

Membrane localization of H+ and HCO3- transporters in the rat pancreatic duct.

The pancreatic duct secretes alkaline fluid that is rich in HCO3- and poor in Cl-. The molecular mechanisms that mediate ductal secretion and are responsible for the axial gradients of Cl- and HCO3- along the ductal tree are not well understood because H+ and HCO3- transport by duct cells have not been characterized or localized. To address these questions, we microdissected the intralobular, main, and common segments of the rat pancreatic duct. H+ and HCO3- transporters were characterized and localized by following intracellular pH while perfusing the bath and the lumen of the ducts. In intralobular ducts, Na(+)-dependent and amiloride-sensitive recovery from acid load in the absence of HCO3- was used to localize a Na+/H+ exchanger to the basolateral membrane (BLM). Modification of Cl- gradients across the luminal (LM) and BLM in the presence of HCO3- showed the presence of Cl-/HCO3- exchangers on both membranes of intralobular duct cells. Measurement of the effect of Cl- on one side of the membrane on the rate and extent of pHi changes caused by removal and addition of Cl- to the opposite side suggested that both exchangers are present in the same cell. In the presence of HCO3-, intralobular duct cells used three separate mechanisms to extrude H+: (a) BLM-located Na+/H+ exchange, (b) Na(+)-independent vacuolar-type H+ pump, and (c) BLM-located, Na(+)-dependent, amiloride-insensitive, and 4',4'-diisothiocyanatostilbene-2,2'-disulfonic acid sensitive mechanism, possibly a Na(+)-dependent HCO3- transporter. The main and common segments of the duct displayed similar mechanisms and localization of H+ and HCO3- transporters to the extent studied in the present work. In addition to the transporters found in intralobular ducts, the main and common ducts showed Na+/H+ exchange activity in the LM. Three tests were used to exclude a significant luminal to basolateral Na+ leak as the cause for an apparent luminal Na+/H+ exchange in an HCO3- secreting cells: (a) addition of amiloride and removal of Na+ from the LM had a profound effect on Na+/H+ exchange activity on the BLM and vice versa; (b) inhibition of all transporters in the BLM by bathing the duct in the inert hydrocarbon Fluorinert FC-75 did not prevent cytosolic acidification caused by removal of luminal Na+; and (c) luminal Na+ did not activate the basolateral Na(+)-dependent HCO3- transporter. An Na(+)-independent, bafilomycin-sensitive H+ pumping activity was marginal in the absence of HCO3-.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

Distinguishing between the catalytic potential and apparent expression of tyrosinase activities.

Assays were developed to investigate the catalytic potential and apparent expression of tyrosinase activities. Tyrosine hydroxylase activity determined with cell lysates (in vitro), entire fixed cells (postfixation), or intact living cells (in situ), and 3,4-dihydroxyphenylalanine oxidase assayed spectrophotometrically or by 3,4-dihydroxyphenylalanine staining on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, demonstrated the following results: 1) The in situ assay displayed reduced tyrosine hydroxylase activity in all three tyrosinase-positive oculocutaneous albino (OCA) lines except for Chediak-Higashi Syndrome melanocytes, which displayed normal activity; 2) The in vitro assay had comparable activity of tyrosinase-positive OCA melanocytes as controls, except for one tyrosinase-positive OCA cell line, which demonstrated increased activity; 3) The postfixation assay, compared with the in situ assay, had elevated activity (ie. normalization) of tyrosinase in OCA cells but reduced activity in controls; 4) The spectrophotometric assay for 3,4-dihydroxyphenylalanine oxidase activity correlated very well with the tyrosine hydroxylase activity determined by the in vitro assay; 5) sodium dodecyl sulfate-polyacrylamide gel electrophoresis of melanocyte lysates either stained with 3,4-dihydroxyphenylalanine or immunoblotted with anti-tyrosinase detected abnormal tyrosinase bands in the Chediak-Higashi Syndrome and one line of tyrosinase positive OCA melanocytes, and both lines had release of tyrosinase into the growth media. In conclusion, the selection and combination of these tyrosinase assays would be informative for differentiation and characterization of human albinism.

Albinism, Oculocutaneous↗
PubMed1994

TGF-beta 1 gene expression in cultured human keratinocytes does not decrease with biologic age.

The biologic activity of cultured epithelial grafts is believed to diminish with increasing cellular age. Therefore, keratinocytes from young donors are used preferentially in the production of cultured allografts for wound treatment. However, the impact of biologic age on cytokine gene expression by human keratinocytes has not been previously investigated. In this study, transforming growth factor-beta 1 (TGF-beta 1) gene expression in human keratinocytes derived from normal foreskins of males ranging in age from 7 months to 82 years was analyzed. Keratinocytes were harvested from fresh specimens and cultivated in vitro on 3T3 fibroblast feeder layers through second passage. The cells were analyzed both qualitatively and semiquantitatively for TGF-beta 1 gene expression using three separate techniques: in situ hybridization, Northern hybridization, and competitive polymerase chain reaction. By in situ hybridization, the signal representing TGF-beta 1 transcript was detected in cells in all layers of the stratified cultures, and immunohistochemical staining for TGF-beta 1 protein was equally intense in all layers. Northern blots of total RNA extracted from the cultivated cells showed no decrease in band density with increasing biologic age. Likewise, no decrease in TGF-beta 1 mRNA levels with biologic age was observed using a semiquantitative polymerase chain reaction assay. These results indicate that the potential for TGF-beta 1 gene expression in cultured foreskin keratinocytes does not decline with increasing cellular age. The findings imply that the clinical performance of cultured grafts, at least as it relates to the elaboration of this growth factor, may not be significantly altered by the biologic age of the keratinocyte donor.

Adult↗
PubMed1994

Human TRP-1 has tyrosine hydroxylase but no dopa oxidase activity.

Human TRP-1 has been immunopurified from normal human melanocytes cultured from black neonatal subjects and used to investigate the catalytic function of TRP-1 for the two substrates, L-tyrosine and L-DOPA. Immunopurified TRP-1 did not demonstrate DOPA staining on SDS/PAGE nor DOPA oxidase (DO) activity with either routine or modified assays. The purified TRP-1 also demonstrated no tyrosine hydroxylase (TH) activity using the routine Pomerantz assay. However, there was apparent TH activity exhibited by immunopurified TRP-1 under conditions with low tyrosine concentration (< or = 0.8 microCi/ml of 3H-tyrosine), prolonged incubation time (i.e., overnight) and in the absence of the cofactor L-DOPA. Using these latter specific conditions, TH activity was also detected in cell lysates from a tyrosinase-negative albino melanocyte line which exhibited no TH activity with the routine Pomerantz assay. In addition, TH activity under low substrate assay conditions was not exhibited in a melanocyte line derived from a TRP-1 deficient, Brown albino individual. However, the absence of TH in this Brown albino cell line could be compensated for by the addition of L-DOPA to the assay. These results suggested that TRP-1 has some tyrosine hydroxylase but no DOPA oxidase activity. We propose that one function of TRP-1 is to modulate tyrosinase activity by making DOPA available as a cofactor to perpetuate the initial steps in melanogenesis.

Cells, Cultured↗
PubMed1994

Transport and interaction of nitrogen oxides and NO2 with CO2-HCO3- transporters in pancreatic acini.

Recently, we showed that NO2- increases gap junction (GJ) permeability and synchronizes intracellular Ca2+ concentration oscillations in pancreatic acini (Loessburg et al., J. Biol. Chem. 268: 19769-19775, 1993). NO2- is also an end product of nitric oxide (NO) production and metabolism. Because of the effect of NO2- on GJ permeability and the possible importance of NO2- in NO metabolism and cytotoxicity, we used pancreatic acinar cells and intracellular pH (pHi) measurements to study the interaction of nitrogen oxides and NO2- with cellular proteins. Exposing cells to NO2- resulted in a concentration-dependent cytosolic acidification. The acidification did not require the transport of NO2- and was not mediated by diffusion of HNO2. Because the acidification was prevented by CO2-HCO3- and inhibition of carbonic anhydrase, it is possible that other nitrogen oxides present in a solution containing NO2- enter the cells by diffusion and interact with OH- or H2O to stably acidify the cytosol. NO2- itself is shown to be transported by the HCO3- transporters present in the plasma membrane. Thus manipulation of the cellular Cl- gradient and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) were used to show Cl-/NO2- exchange, whereas stimulation of external Na(+)-dependent amiloride-insensitive and DIDS-sensitive pHi increase in acidified cells was used to demonstrate a Na(+)-(NO2-)n cotransport. Hence NO2- can be a convenient substitute for HCO3- when studying HCO3- transport in an open system. The studies also show that cellular levels of nitrogen oxides and NO2- can be modulated by the cellular HCO3(-)-buffering system.(ABSTRACT TRUNCATED AT 250 WORDS)

Acids↗
PubMed1994

[Study on difference of Za (miscellaneous) and Cu (abrupt)].

So far, the contention on the title of Zhang Zhongjing's work, either Shang Han Za Bing Lun or Shang Han Cu Bing Lun, is still unsettled. The present paper probes this problem by several approaches, including the provenance of now extant Zhongjing's book, the stemming of the mystery Za and Cu, the evolution of title of Zhongjing's book before both Song Dynasties; the origin and development of miscellaneous disease; semantic of both Za and Cu. On the above basis, a comprehensive study has come to the conclusion that the "wrong character" hypothesis mentioned by Song scholars is groundless, and the correct title for Zhongjing's work should read Shan Han Cu Bing Lun. The paper also discusses logically the development of nosological system in TCM.

China↗
PubMed1994

[Modified en bloc resection procedure for malignant tumor of the shoulder girdle].

En bloc resection of the shoulder girdle (Tikhoff-Linberg procedure) is a valuable limb-sparing surgical technique for bony and soft tissue malignant tumor in and around the proximal humerus and shoulder girdle. Significant amount of hand-elbow function is maintained with the procedure, and long time results in the treatment of neoplasms by this technique was found to be the same as by the method of a forequarter amputation or the shoulder disarticulation in the English literature. Four cases with malignant tumor in the shoulder girdle was treated with this modified method in our department. The average follow-up was 8 years. One case died due to another kind of tumor 6 years after operation, and other 3 cases are alive and disease free.

Adult↗
PubMed1994

[Effect of GG on the occurrence of tongue retropulsion--an experimental observation of electromyography].

To evaluate the effect of GG activity on tongue backwardness during sleep, we conducted experiments on 16 rabbits. The electric activities of the muscles involved in tongue motion were recorded in different phases (wakefulness, presnoring and snoring) with unipolar electrodes inserted into the muscles under direct vision. The observations demonstrated that GG played a more active role in the occurrence of tongue backwardness during sleep.

Animals↗
PubMed1994

[The use of internal fixation in the treatment of spinal tuberculosis].

We have applied debridement with grafting and internal fixation techniques (including Luque and Dick procedures) in the treatment of tuberculosis of the spine in the past few years. Analysis of 18 patients showed good prognosis after the Luque or Dick procedure. The patients are able to walk around within a few days after operation. One to five years of follow-up showed no recurrence or kyphotic deformity.

Adolescent↗