PubMed Health⌕ Search

Biomedical subjects

H F Wang

Publications and source records attributed to H F Wang.

At least 37 records · Page 2Linked to original sources

Rapid analysis of terpene lactones in extract of Ginkgo biloba L. by high performance liquid chromatography.

A new rapid analytical method was developed for the quantification of terpene lactones (bilobalide(BB) and ginkgolide A, B, C, J) in extract of Ginkgo biloba L. (EGb) using a liquid-liquid solvent extraction procedure followed by high performance liquid chromatography. EGb was dissolved in 30% ethanol and extracted with ether. After evaporation, the residue was then determined by HPLC on a C18 column with methanol-water-orthophosphoric acid (25:75:0.1, V/V) as eluent and refractive index (RI) detection. Results showed that the excellent sample clean-up procedure is more simple and specific, and saves more time (less than 20 min) than any other methods that have been reported, and also leads to high recoveries (> 99.0%) and low RSDs (< 2.0%). The reproducible method is regarded to be very useful for evaluating the quality of extract of Ginkgo biloba L.

Chromatography, High Pressure Liquid↗

Cytochrome P450 3A degradation in isolated rat hepatocytes: 26S proteasome inhibitors as probes.

Mechanism-based inactivation of liver microsomal cytochromes P450 3A (CYP 3A, P450s 3A) in vivo and/or in vitro, via heme modification of the protein, results in accelerated proteolytic degradation of the enzyme that is preceded by the ubiquitination of the protein, thereby implicating the ubiquitin-ATP-dependent 26S proteasomal system. In this study, this involvement is confirmed with the use of the proteasomal inhibitors aclarubicin and MG-132 as probes, in isolated rat hepatocytes treated with the P450 3A mechanism-based inactivator, 3,5-dicarbethoxy-2,6-dimethyl-4-ethyl-1, 4-dihydropyridine (DDEP). In addition, the findings reveal that during the course of this proteolysis, the endoplasmic reticulum (ER)-anchored DDEP-inactivated P450 3A is translocated from the ER to the cytosol in a brefeldin A-insensitive manner.

Aclarubicin↗

Increased uptake and transport of cholera toxin B-subunit in dorsal root ganglion neurons after peripheral axotomy: possible implications for sensory sprouting.

In the present study we show that, in contrast to the rat, injection of cholera toxin B-subunit (CTB) into the intact sciatic nerve of Macaca mulatta monkey gives rise to labelling of a sparse network of fibers in laminae I-II of spinal cord and of some mainly small dorsal root ganglion (DRG) neurons. Twenty days after sciatic nerve cut, the percentage of CTB-positive lumbar 5 (L5) DRG neuron profiles increased from 11% to 73% of all profiles. In the spinal cord, a marked increase in CTB labelling was seen in laminae I, II, and the dorsal part of lamina III. In the rat L5 DRGs, 18 days after sciatic nerve cut, the percentage of CTB- and CTB conjugated to horseradish peroxidase (HRP)-labelled neuron profiles increased from 45% to 81%, and from 54% to 87% of all neuron profiles, respectively. Cell size measurements in the rat showed that most of the CTB-positive neuron profiles were small in size after axotomy, whereas most were large in intact DRGs. In the rat spinal dorsal horn, a dense network of CTB-positive fibers covered the whole dorsal horn on the axotomized side, whereas CTB-labelled fibers were mainly seen in laminae III and deeper laminae on the contralateral side. A marked increase in CTB-positive fibers was also seen in the gracile nucleus. The present study shows that in both monkey and rat DRGs, a subpopulation of mainly small neurons acquires the capacity to take up CTB/CTB-HRP after axotomy, a capacity normally not associated with these DRG neurons. These neurons may transganglionically transport CTB and CTB-HRP. Thus, after peripheral axotomy, CTB and CTB-HRP are markers not only for large but also for small DRG neurons and, thus, possibly also for both myelinated and unmyelinated primary afferents in the spinal dorsal horn. These findings may lead to a reevaluation of the concept of sprouting, considered to take place in the dorsal horn after peripheral nerve injury.

Animals↗

Distribution of transganglionically labelled soybean agglutinin primary afferent fibres after nerve injury.

The distribution of the retrogradely-transganglionically transported lectin soybean agglutinin (SBA) and of SBA conjugated to horseradish peroxidase (SBA-HRP) has been examined in the L4-5 dorsal root ganglia, lumbar spinal cord and gracile nucleus at 2, 6 and 14 weeks after sciatic nerve transection and ligation. Cell size analysis showed there were no changes in the mean area of labelled DRG profiles after injury. In the spinal cord, terminal labelling was restricted to laminae I and II with no evidence of labelling in novel territories such as the deeper laminae after injury. At 2 weeks, the labelling on the injured side was similar in distribution and intensity to that of the contralateral, uninjured side. At 6-14 weeks the labelling on the injured side was significantly weaker as compared to the contralateral side, but not completely depleted. In the gracile nucleus, at all survival times, an increased distribution and amount of labelling was seen which may reflect sprouting of C and A-delta fibres. These results suggest that SBA is a useful tracer to study the effects of nerve injury on the central terminals of axotomised afferents terminating in laminae I-II and that C-fibres appear not to sprout outside their normal laminar distribution in the dorsal horn after injury.

Animals↗

Anterograde transport of horseradish-peroxidase conjugated isolectin B4 from Griffonia simplicifolia I in spinal primary sensory neurons of the rat.

Anterograde transport of the isolectin B4 from Griffonia simplicifolia I (B4) conjugated to horseradish peroxidase (HRP) was investigated in rat somatic and visceral primary sensory neurons at different spinal levels. Injection of B4-HRP into the L5 dorsal root ganglion (DRG) resulted in labelling in the sural nerve, but not in the gastrocnemius nerves. Free nerve endings and lanceolate-like nerve endings were labelled in the lateral hindpaw skin. Labelled fibres were also observed in the greater splanchnic nerve following B4-HRP injection into the T10-11 DRGs. Electron microscopic examination of the labelled nerves showed that B4-HRP labelled exclusively unmyelinated axons. In the spinal cord, labelling was observed in the superficial dorsal horn, and additionally, although much more sparse, in the medial and lateral collateral projections following injections into the T10-11 DRGs. The results suggest that B4-HRP should be a suitable anterograde tracer of unmyelinated cutaneous and splanchnic but not muscle primary afferent fibres.

Animals↗

Expression of type VI adenylyl cyclase in the central nervous system: implication for a potential regulator of multiple signals in different neurotransmitter systems.

The aim of this study was to investigate the distribution of a calcium-inhibitable adenylyl cyclase type VI (type VI AC) in the central nervous system using an antiserum directed against the N-terminus of type VI AC. Our results indicate that type VI AC immunoreactivity is generally expressed in many brain regions with different levels of intensity. Most interestingly, the majority of the detected type VI AC immunoreactivity is present in cells of neuronal phenotype. Double immunostaining of type VI AC and markers of various neurotransmitter systems suggest that type VI AC might participate in regulation of the classical neurotransmitter systems and therefore appeared to play a very important role in the central nervous system.

Adenylyl Cyclases↗

Retrograde and transganglionic transport of horseradish peroxidase-conjugated cholera toxin B subunit, wheatgerm agglutinin and isolectin B4 from Griffonia simplicifolia I in primary afferent neurons innervating the rat urinary bladder.

In the present study, we investigated and compared the ability of the cholera toxin B subunit, wheat germ agglutinin and isolectin B4 from Griffonia simplicifolia I conjugated to horseradish peroxidase, to retrogradely and transganglionically label visceral primary afferents after unilateral injections into the rat urinary bladder wall. Horseradish peroxidase histochemical or lectin-immunofluorescence histochemical labelling of bladder afferents was seen in the L6-S1 spinal cord segments and in the T13-L2 and L6-S1 dorsal root ganglia. In the lumbosacral spinal cord, the most intense and extensive labelling of bladder afferents was seen when cholera toxin B subunit-horseradish peroxidase was injected. Cholera toxin B subunit-horseradish peroxidase-labelled fibres were found in Lissauer's tract, its lateral and medial collateral projections, and laminae I and IV-VI of the spinal gray matter. Labelled fibres were numerous in the lateral collateral projection and extended into the spinal parasympathetic nucleus. Labelling from both the lateral and medial projections extended into the dorsal grey commissural region. Wheat germ agglutinin-horseradish peroxidase labelling produced a similar pattern but was not as dense and extensive as that of cholera toxin B subunit-horseradish peroxidase. The isolectin B4 from Griffonia simplicifolia I-horseradish peroxidase-labelled fibres, on the other hand, were fewer and only observed in the lateral collateral projection and occasionally in lamina I. Cell profile counts showed that a larger number of dorsal root ganglion cells were labelled with cholera toxin B subunit-horseradish peroxidase than with wheat germ agglutinin- or isolectin B4-horseradish peroxidase. In the L6-S1 dorsal root ganglia, the majority (81%) of the cholera toxin B subunit-, and almost all of the wheat germ agglutinin- and isolectin B4-immunoreactive cells were RT97-negative (an anti-neurofilament antibody that labels dorsal root ganglion neurons with myelinated fibres). Double labelling with other neuronal markers showed that 71%, 43% and 36% of the cholera toxin B subunit-immunoreactive cells were calcitonin gene-related peptide-, isolectin B4-binding- and substance P-positive, respectively. A few cholera toxin B subunit cells showed galanin-immunoreactivity, but none were somatostatin-, vasoactive intestinal polypeptide-, or neuropeptide Y-immunoreactive or contained fluoride-resistant acid phosphatase. The results show that cholera toxin B subunit-horseradish peroxidase is a more effective retrograde and transganglionic tracer for pelvic primary afferents from the urinary bladder than wheat germ agglutinin-horseradish peroxidase and isolectin B4-horseradish peroxidase, but in contrast to somatic nerves, it is transported mainly by unmyelinated fibres in the visceral afferents.

Animals↗

Transganglionic transport of the lectin soybean agglutinin (Glycine max) following injection into the sciatic nerve of the adult rat.

The lectin soybean agglutinin (SBA) from Glycine max binds to small-sized dorsal root ganglion cells and their central terminals in the superficial dorsal horn of the spinal cord. Here we investigated the ability of SBA and SBA conjugated to horseradish peroxidase (SBA-HRP) to trace thin calibre afferents into the spinal cord from a peripheral nerve. Following injection into the sciatic nerve, labelled cells in the dorsal root ganglion were predominantly small-sized but some medium-sized cells were also labelled. Colocalization studies of transported SBA with various neuronal markers showed that all neurons that transported SBA-HRP showed SBA binding, indicating high uptake specificity for the conjugate. 15% were immunoreactive for RT97 indicating that some axons were myelinated, and 54% also expressed binding sites for isolectin B4 from Griffonia simplicifolia, a selective marker for a subpopulation of unmyelinated afferents. With regard to neurotransmitter content, 43% of the SBA cells contained calcitonin gene-related peptide, 33% contained substance P and 2.5% somatostatin. In addition, 3% contained carbonic anhydrase. Centrally, injection of SBA in the sciatic nerve resulted in labelled terminals in somatotopically appropriate regions of laminae I-II of the dorsal horn, and in the gracile nucleus. A few neurons in the dorsal horn were labelled indicating that some transneuronal transport of SBA had occurred. The results show that SBA can be used as a transganglionic tracer to label fine calibre primary afferents that project to laminae I-II of the spinal cord and the gracile nucleus. It appears to label more afferents than isolectin B4, including also a subpopulation of myelinated afferents.

Age Factors↗

Differentiation and grouping of isolates of the Ganoderma lucidum complex by random amplified polymorphic DNA-PCR compared with grouping on the basis of internal transcribed spacer sequences.

Laccate polypores of the Ganoderma lucidum species complex are widespread white rot fungi of economic importance, but isolates cannot be identified by traditional taxonomic methods. Parsimony analysis of nucleotide sequences from the internal transcribed spacers (ITS) of the ribosomal gene (rDNA) distinguished six lineages in this species complex. Each ITS lineage may represent one or more putative species. While some isolates have identical ITS sequences, all of them could be clearly differentiated by genetic fingerprinting using random amplified polymorphic DNA (RAPD). To investigate the suitability of RAPD markers for taxonomic identification and grouping of isolates of the G. lucidum complex, RAPD fragments (RAPDs) were used as phenotypic characters in numerical and parsimony analyses. Results show that data from RAPDS do not distinguish the same clades as ITS data do. Groupings based on analysis of RAPD data were very sensitive to the choice of the grouping method used, and no consistent grouping of isolates could be proposed. However, analysis with RAPDs did resolve several robust terminal clades containing putatively conspecific isolates, suggesting that RAPDs might be helpful for systematics at the lower taxonomic levels that are unresolved by ITS sequence data. The limitations of RAPDs for systematics are briefly discussed. The conclusion of this study is that ITS sequences can be used to identify isolates of the G. lucidum complex, whereas RAPDs can be used to differentiate between isolates having identical ITS sequences. The practical implications of these results are briefly illustrated.

Base Sequence↗

Glutathione S-transferase pi facilitates the excretion of arsenic from arsenic-resistant Chinese hamster ovary cells.

We have previously demonstrated that glutathione S-transferase pi (GST pi) is overexpressed in SA7 cells, an arsenic resistant cell line derived from Chinese hamster ovary (CHO) cells. Our present results show that SA7 cells accumulate less arsenic than parental CHO cells and partially revertant SA7N cells. The lower levels of arsenic accumulation in SA7 cells resulted from their faster excretion rates. However, the excretion of arsenic from SA7 cells was significantly inhibited by the GST inhibitors ethacrynic acid and Cibacron blue. Furthermore, when GST pi levels in SA7N cells were re-elevated by zinc sulfate pretreatment, arsenic accumulation decreased and arsenic excretion increased to levels similar to those in SA7 cells. These results suggest that GST pi can facilitate the excretion of arsenic. Such facilitation by GST pi is unlikely to be associated with multi-drug resistant P-glycoprotein, since no overexpression of P-glycoprotein was detected in SA7N and SA7 cells.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Glutathione S-transferase pi in an arsenic-resistant Chinese hamster ovary cell line.

A glutathione S-transferase (GST) was purified from an arsenic-resistant Chinese hamster ovary cell line, SA7. The SA7 GST was shown to catalyse the conjugation of glutathione and ethacrynic acid, a specific substrate for Pi class GST. Its N-terminal amino-acid sequence has 80% identical residues to that of rat GST P and human GST pi. Thus, the GST purified from SA7 cells belongs to the Pi family. Treatment with Cibacron Blue or ethacrynic acid, which are GST inhibitors, significantly decreased the resistance of SA7 cells to sodium arsenite. On the other hand, pretreatment of SA7N cells, a partial revertant of SA7 cells, with sublethal doses of sodium arsenite, cadmium acetate or zinc sulphate resulted in re-elevation of GST activities and the cells regained the arsenic resistance. The regained arsenic resistance was well correlated with the levels of GST pi which were induced dose-dependently by zinc sulphate. Heat-shock treatment (45 degrees C for 10 min) did not increase GST pi expression or arsenic resistance of SA7N cells. The results indicate that GST pi is possibly involved in the mechanism of arsenic detoxification.

Amino Acid Sequence↗

Radiation-protective and platelet aggregation inhibitory effects of five traditional Chinese drugs and acetylsalicylic acid following high-dose gamma-irradiation.

High doses of 60Co radiation (4.0-8.0 Gy) in mice, rats and rabbits caused increases in rate of platelet aggregation during the first 5 days after irradiation. The inhibitory effects of the extracts of five Chinese drug plants and acetylsalicylic acid on rate of platelet aggregation were observed in both in vitro and in vivo tests, averaging 23-53% in vitro and 46-69% in vivo. Antiradiation tests on mice vs. 7.5-8.0 Gy of gamma-radiation, using the plant extracts and acetylsalicylic acid as protective agents, increased survival rates by 8-50% for the plant extracts and 35% for acetylsalicylic acid.

Animals↗

Radiotracer study on mancozeb residues in tomato plants.

A study on the persistence, residues and residue distribution of mancozeb in tomato plant-soil system, as well as effects of cooking on transformation of residual mancozeb were carried out with radiotracer technique. Half-life values for total mancozeb residues on tomato leaves and in soil were observed to be 9.5 and 7.6 days, respectively. The results also suggested that mancozeb residues in soil were readily uptaken by plants and leach to soil layer as deep as 28cm within two weeks, although over 90% of the residues remained in the upper layer (0-6cm) of the soil. Translocation of the residues of mancozeb, especially the water soluble metabolites including ETU, throughout the plants could be one of the factors responsible for the contamination of tomato fruits. Deposit of sprayed mancozeb on the fruit skin was another important factor leading to the contamination of fruits. Fruit skin contained higher levels of residues than the pulp. Washing with water could remove more than 50% of the residues on the skin. It was also found that 20-30% of the residual mancozeb degraded to ETU during cooking process. The tomatoes should be washed to reduce the residues of the fungicide and ETU in food.

Autoradiography↗

[Influence of several Chinese drugs on the glycogen synthesizing function of primary cultured adult rat hepatocytes].

The influence of several Chinese drugs on the glycogen synthesizing function of normal and carbon tetrachloride-injured hepatocytes was investigated. Hepatocytes prepared from rats fasted for 16 h were incubated with 50 mmol/L glucose. Glycogen content was determined 30 and 90 min after incubation with normal and CCl4-injured cells, respectively. Insulin was used as positive control which increased glycogen content and the data coincided with that in the literature. The following results were obtained (1) Biphenyl-dimethyl-dicarboxylate (BDD) in 100 micrograms/ml concentration increased glycogen content of normal hepatocytes by 88%. It protected cells against CCl4-injury:BDD 10 micrograms/ml remarkably decreased CCl4-induced reduction of glycogen and 100 micrograms/ml showed complete protection. (2) Tremella poly-saccharide slightly increased glycogen content in normal cells, but in a concentration of 100 micrograms/ml it decreased CCl4-induced reduction of glycogen significantly. (3) Low concentration of norcatharidin (10 micrograms/ml) increased glycogen content of normal cells, but at 100 micrograms/ml concentration this effect disappeared. Furthermore, it intensified the toxic effect of CCl4 on glycogen at 10-100 micrograms/ml. (4) CL1500 (100 micrograms/ml) increased glycogen content of normal cells, but it intensified CCl4-injury effect on glycogen at the same concentration.

Animals↗

The effect of copper IUD on microcirculation of blood flow in rabbit uterus.

A survey of the rabbit's normal microcirculation of blood flow volume in the endometrium and myometrium with or without a copper intrauterine device (IUD) was made. It was found through statistical analyses that the IUD stimulated the endometrium so that microcirculation of blood flow volume in the endometrium increased significantly, but it did not change the microcirculation of blood flow volume in the myometrium and in the opposite uterus.

Animals↗