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Biomedical subjects

J Huang

Publications and source records attributed to J Huang.

At least 559 records · Page 31Linked to original sources

[Experimental studies on immunocompetence of lophanthus roots].

The effect of Lopanthus roots extract on the HI humoral immunity and cell-mediated immunity in rat was investigated in this study. The result showed that the Lophanthus roots extract could enhance liver index in rat. By Lophanthus roots extract at the dosage of 2.50 g/kg in rat, the Immunoglobulin G(IgG), immunoglobulin A(IgA) and immunoglobulin M (IgM) were increased, C3 complenment exhanced and have a highly signifcance compared to the cation by the gingseng veinforce pill (P < 0.01). The Lophanthus roots extract could mildly increased effect of rosette formation and lymphocyte transformation, but no statistic sighnificance was found.

Adjuvants, Immunologic↗

[Characteristics of virus growth factor (VGF) encoded by vaccinia virus Tian Tan strain].

Vaccinia virus growth factor (VGF) is encoded by an early gene located near the border between the terminal inverted repeats and the internal unique region of the viral genome. It has been suggested that secreted VGF may serve to stimulate the metabolism of neighboring uninfected cells, thereby increasing the capacity of these cells to support viral replication. In this study, we cloned and sequenced the VGF gene of vaccinia virus strain Tian Tap. The analytical results indicated that VGF of strain Tian Tan possessed the core sequences of Epidermal growth factor superfamily, and more closely related to that of strain WR than to those of other orthopoxvirus strains. Overall deduced amino acid divergence of 4.3% was observed between Tian Tan and WR VGF protein, while divergences ranged from 10% to 20% compared to other orthopoxviruses strains. Most of the amino acid substitutions were found within signal peptide and transmembrane regions, but were not present in secreted ligand domain. It was striking that the amino acid sequence deduced from the ORF TB22L which located within the right terminal region of genome was identical to VGF protein amino acids 67-140, suggesting that ORF TB22L may be derived from the seconed copy of VGF gene of strain Tian Tan.

Intercellular Signaling Peptides and Proteins↗

[Determination of degradation level of antifungal agent in mice serum by high performance liquid chromatography].

L-4-oxalysinyl-norvalinyl-N3-4-methoxyfumaroyl-L-2,3-diaminopropanoic acid (I-677-Nva-FMDP) is a new tripeptide synthesized in our group. This peptide exhibits potent anticandida Albicans activity. In the presence of serum, the antifun gal activity of I-677-Nva-FMDP decreases after incubating over 1h. In order to investigate the relationship between the degradation and antifungal activity of I-677-Nva-FMDP, the concentrations of I-677-Nva-FMDP and N3-methoxyfumaroyl-L-2,3-diaminopropanoic acid (FMDP) were determined at different incubating time in mice serum. The incubating time range was from 0 to 300 min and the incubating samples were measured at intervals of 30 min. For this measurement reversed-phase HPLC was used and the mobile phase was composed of 8% methanol and 0.1% trifluoroacitic acid-triethyl amine buffer (pH 3). In this condition the retention time of FMDP and I-677-Nva-FMDP were 3.9 min and 16.5 min respectively. Methanol was superior to other reagents for the removal of protein from the incubating medium and there was not any interference peak before the retention time of I-677-Nva-FMDP. A decrease in concentration of I-677-Nva-FMDP was observed from 0 min to 180 min and no I-677-Nva-FMDP could be detected at 210 min whereas the concentration of FMDP incresed from 30 min to 2h and reached a maximum at 120 min. The results showed that the half life (t1/2) of I-677-Nva-FMDP was 70 min. This result coincided with the antifungal test in vitro.

Animals↗

Avermectin B1a binds to high- and low-affinity sites with dual effects on the gamma-aminobutyric acid-gated chloride channel of cultured cerebellar granule neurons.

Avermectin B1a (AVM B1a), a widely used insecticide and acaricide, is reported to both activate and inhibit gamma-aminobutyric acidA (GABA(A)) receptor function in mammalian brain. This study attempts to resolve these seemingly contradictory results by examining the binding properties of AVM B1a and its effects on the GABA-gated chloride channel with primary cultures of rat cerebellar granule neurons as a model system. Specific binding of [3H]AVM B1a in intact neuron cultures is time- and concentration-dependent and is displaceable by AVM analogs. Scatchard analysis of [3H]AVM B1a binding reveals high- and low-affinity sites with K(D) values of 5 and 815 nM, respectively. AVM B1a alters the binding of [3H]ethynylbicycloorthobenzoate at the noncompetitive blocker site in a biphasic manner; activation is evident with 10 to 300 nM AVM B1a after 5 to 10 min incubation and inhibition with an IC50 of 866 nM after 60 min incubation. Consistent with this observation, 36Cl- influx is stimulated by AVM B1a at 3 to 100 nM and inhibited at 1 to 3 microM. GABA-stimulated 36Cl- influx is completely blocked by both [3H]ethynylbicycloorthobenzoate and 12-ketoendrin (two GABA-gated chloride channel blockers) and AVM B1a at 1 to 1.5 microM. Also, 36Cl- influx induced by AVM B1a at 10 nM is suppressed by the two channel blockers. Thus, AVM B1a binds to two different sites in the GABA-gated chloride channel with dual effects, i.e., activating the channel on binding to the high-affinity site and blocking it on further binding to the low-affinity site.

Animals↗

Cellular retinaldehyde-binding protein is expressed by oligodendrocytes in optic nerve and brain.

Cellular retinaldehyde-binding protein (CRALBP) is abundant in the retinal pigment epithelium and Müller glial cells of the retina, where it forms complexes with endogenous 11-cis-retinoids. We examined the distribution of CRALBP in extraretinal tissues using polyclonal antibodies (pAb) and monoclonal antibodies (mAb). A protein was detected by immunoblot analysis in extracts of bovine and rat brain and optic nerve but not in several other tissues. This protein had electrophoretic, chromatographic, and retinoid-binding properties identical to those of CRALBP from bovine retina. Comparison of the masses of tryptic peptides and of partial amino acid sequences derived from brain and retinal CRALBP indicated that the two proteins are probably identical. Immunoperoxidase cytochemistry and double labeling immunofluorescence revealed CRALBP(+) cells in brain that resembled oligondendrocytes and not astrocytes, microglial cells, or pinealocytes. In 11-day-old rat brain, approximately 11% of the CRALBP(+) cells were labeled with the Rip antibody, a marker for oligodendroglia. In developing rat optic nerve, the temporal appearance of CRALBP(+) cells corresponded to that of oligodendrocytes and not that of astrocytes. In adult rat and mouse optic nerves, the CRALBP(+) somata showed the same distribution as oligodendrocytes. No endogenous retinoids were associated with CRALBP isolated from dark-dissected adult bovine brain. The results suggest that CRALBP has functions in addition to retinoid metabolism and visual pigment regeneration.

Amino Acid Sequence↗

[Chromosomal aberrations induced in human spermatozoa by mitomycin C].

Human spermatozoa were exposed to Mitomycin C (MM() at the concentrations of 7.5, 15, 3O micrograms/ml for 1 hour respectively before interspecific in vitro fertilization with zona-free hamster ova. The first cleavage metaphases was analyzed to investigate MMC-induced effects on human spermatozoa. The results showed a distinct dose-response relationship in the induction of chromosomal aberrations. The chromatid aberrations were the predominant types in MMC-induced changes. Some chromosome-type aberrations were also observed. This suggested that the induced effects of MMC on human spermatozoa were some different from that of ultraviolet light-like clastogens on somatic cells. 36.89% of MMC-induced aberrations were rejoining-type. This indicated that the DNA repair systems present in golden hamster oocytes were effective in repairing MMC-induced DNA lesions in human spermatozoa.

Animals↗

[Expression of glutathione S-transferase-pi in operative specimens as marker of chemoresistance in patients with ovarian cancer].

OBJECTIVE: To study the relationship between the expression of glutathione S-transferase-pi (GST-pi) in operative specimens and chemoresistance in patients with ovarian cancer. METHODS: The expression of GST-pi in 87 epithelial ovarian cancer tissues and 30 normal ovarian epithelial tissues was determined with labelled streptavidin biotin method (LSAB). All the patients had not received chemotherapy before operation. We used Chi-Square and Cox-Mantel test to analyze the relativity between the expression of GST-pi and clinical pathological data, chemotherapeutic response, prognosis in patients with ovarian cancer. RESULTS: (1) 59 (67.8%) of 87 ovarian cancer tissue were demonstrated to be positive expression with GST-pi, but all 30 normal ovarian epithelial tissue were negative. (2) There was no direct correlation between the expression of GST-pi and clinical pathological data. (3) 43 (43/59) of GST-pi positive cases were chemoresistant, while only 3 (3/28) of GST-pi negative ones were chemoresistant. (4) The difference in the chemotherapeutic response between the two groups was obviously significant (P < 0.005). The survival period of the patients with GST-pi positive expression was also obviously shorter than that of those with GST-pi negative expression (P = 0.004). CONCLUSION: These results strongly suggest that GST-pi expression in epithelial ovarian cancer tissues is closely related to chemoresistance clinically and it may be served as a useful marker to predict the prognosis of patients.

Antineoplastic Agents↗

Sonographic diagnosis of bone tumors.

This paper presents the results of sonographic and radiographic examinations in 48 patients clinically diagnosed as having bone tumors. Sonography revealed bone destruction in all 48 cases, elevated periosteum in 26 cases and soft tissue mass in 34 cases. The results obtained in this series demonstrate that most bone tumors have their characteristic sonographic features such as giant cell tumors, malignant bone tumors, bone cysts, as well as metastatic lesions. Study showed that sonography has equally high accuracy in the diagnosis of these tumors compared with radiography.

Adolescent↗

[The effect of root canal irrigation and variance of apical foramen pressure by use of casing aspiration needle system in vitro].

The authors introduced casing aspiration needle system (CANS) to irrigating root canal. The effect of root irrigation on removing canal debris and the variance of apical foramen pressure on extracted teeth were compared between the conventional irrigation and CANS irrigation. The results indicated that the effect of root canal irrigation by CANS was much better than the conventional method (P < 0.01). CANS irrigation didn't produce pressure to apical foramen during root canal irrigation. However, in the conventional method, with irrigation needle inserted deeper and deeper in root cancal the pressure of apical foramen gets higher and higher.

Humans↗

hCG-receptor binding and transmembrane signaling.

The technique of site-directed mutagenesis has proven to be quite powerful in elucidating contact sites involved in the interaction of the heterodimeric glycoprotein hormones and their respective seven transmembrane (TM) G protein-coupled receptors. Our laboratory has focused on identification of the minimum core sequences of the alpha and beta subunits required for bioactivity, the minimum length of a conjoined (yoked) single-chain hCG, the amino acid residues on hCG and the LH/CG-receptor (LH/CG-R) responsible for high-affinity binding, and the regions of the receptor that are involved in TM signaling. A number of amino acid residues have been mapped on the alpha and beta subunits of hCG that appear important in receptor binding. When projected onto the crystal structure of HF-treated hCG, these residues, by and large, cluster on one side of the molecule and cover a sizeable surface area, indicating that the hormone-receptor binding interface is rather extensive. Based on mutagenesis studies of several conserved ionizable amino acid residues in the extracellular domain (ECD) of LH/CG-R and a model that we, in collaboration with Drs Lapthorn and Isaacs, have developed for this region based on the crystal structure of porcine ribonuclease inhibitor, a charged region that appears to play an important role in hormone-receptor recognition has been identified. We have also delineated several regions of LH/CG-R that do not appear to participate in hCG binding but are involved in hCG-mediated signaling. These regions are located in the ECD and extracellular loop III just prior to entry into the membrane via TM helices I and VII, respectively, and in TM helices VI and VII. Similarly, a homologous region in the ECD of the FSH receptor, located with ten residues of TM helix I, is important in signaling but not hormone binding. These results suggest that ligand binding and ligand-mediated receptor activation are quasi-distinct, albeit sequential phenomena. Collectively, our mutagenesis and modeling studies, coupled with results from other laboratories, argue for a ligand-induced conformational change of the receptor that may involve a relative reorientation of the TM helices.

Amino Acid Sequence↗

Microtubule-dependent multilobular organization of the nucleus in sensitive and multidrug-resistant L0 leukemia cells.

The relationship between the nuclear morphology and the microtubular organization of L100 and L1000 cells, two vincristine-induced multidrug resistant human acute lymphocytic leukemia cell lines, was examined and compared to that of L0 parental cells. The L0 parental cells contained a round nucleus and the microtubules were evenly distributed throughout the cytoplasm. In contrast, the microtubules of the L100 and L1000 cells were localized between the lobular structures of a multilobulated nucleus. Disassembly of microtubules in L100 and L1000 cells by colchicine resulted in the loss of the multilobulated morphology of the nucleus. While the total cellular content of tubulin of L0 and L100 cells was similar, the content of microtubules of L100 cells was only 55% of that observed in L0 cells. Two, 28 kDa (pI 6.9) and 31 kDa (pI 4.4), microtubule-associated proteins were found to be overexpressed in L100 and L1000 cells. The results indicate that the multilobulated nuclear morphology of L100 and L1000 cells is dependent upon the unique and intact organization of the microtubules; the distinct organization of the microtubules and the multilobular nuclear morphology of the two resistant cell lines may be due to the differential expression of specific microtubule-associated proteins.

Cell Cycle↗

APC mutations in colorectal tumors with mismatch repair deficiency.

We have investigated the influence of genetic instability [replication error (RER) phenotype] on APC (adenomatous polyposis coli), a gene thought to initiate colorectal tumorigenesis. The prevalence of APC mutations was similar in RER and non-RER tumors, indicating that both tumor types share this step in neoplastic transformation. However, in a total of 101 sequenced mutations, we noted a substantial excess of APC frameshift mutations in the RER cases (70% in RER tumors versus 47% in non-RER tumors, P < 0.04). These frameshifts were characteristic of mutations arising in cells deficient in DNA mismatch repair, with a predilection for mononucleotide repeats in the RER tumors (P < 0.0002), particularly (A)n tracts (P < 0.00007). These findings suggest that the genetic instability that is reflected by the RER phenotype precedes, and is responsible for, APC mutation in RER large bowel tumors and have important implications for understanding the very earliest stages of neoplasia in patients with tumors deficient in mismatch repair.

Adenomatous Polyposis Coli↗

Effects of human polymorphonuclear leukocyte elastase upon surfactant proteins in vitro.

Recent evidence has suggested that elastase is released by polymorphonuclear leukocytes (PMN) recruited from the pulmonary microcirculation into the alveoli during acute lung injury. This study was undertaken to test the hypothesis that elastase from PMN (PMN elastase) damages or degrades one or more of the surfactant proteins (SP-A, SP-B and SP-C) of the lung, and thereby alters its function. We attempted to use amounts of PMN elastase and quantities of surfactant that would be plausible in the lungs of patients with ARDS. Surfactant from normal dog lungs (2 mg phospholipid, 200 micrograms protein), and purified SP-A (20 micrograms), SP-B (10 micrograms) and SP-C (10 micrograms) from the surfactant (identified by SDS-PAGE and N-terminal amino acid sequences) were incubated for 4-8 h at 37 degrees C with various amounts (0.25-1.0 U) of human PMN elastase purified by affinity chromatography. SDS-PAGE and amino acid composition analysis of the surfactant as well as of the purified SP-A, SP-B, and SP-C showed that degradation of these proteins progressed with incubation time and with the amount of PMN elastase, and was accompanied by decreases in isopycnic density (g/cm3) and surface adsorption, and increase of surface tension of the surfactant. No effects were observed with heat inactivated PMN elastase (95 degrees C, 30 min) or with PMN elastase in the presence of human alpha-1 protease inhibitor (2 micrograms/microgram elastase). Phospholipid compositions of the surfactant after exposure to PMN elastase were not significantly different from those of the controls, suggesting that SP-A, SP-B, and SP-C play a major role in altering the surfactant properties. SP-A was also degraded by elastase and trypsin from pancreas whereas SP-B and SP-C remained intact, providing a natural surfactant without SP-A. Surface adsorption rate of the SP-A deficient surfactant was lower than that of the control, but was much higher than that of the surfactant with completely degraded SP-A, SP-B, and SP-C, suggesting that hydrophobic SP-B and SP-C are the essential components in enhancing adsorption. We conclude that proteolytic degradation of SP-A, SP-B, and SP-C causes the decrease of surfactant isopycnic density, and is responsible for retarding adsorption resulting in surfactant dysfunction.

Amino Acid Sequence↗

Polymorphism of angiotensin I converting enzyme gene in the older Chinese: linked to ambulatory blood pressure levels and circadian blood pressure rhythm.

This study was undertaken to evaluate the association of polymorphism of angiotensin I converting enzyme (ACE) gene with causal and ambulatory blood pressure in the Chinese population. Genomic DNA was amplified by polymerase chain reaction (PCR) using primers flanking the polymorphic region in intron 16 of the ACE gene. Alleles were detected on agarose gels stained with ethidium bromide. Causal blood pressure was measured in the 133 normotensive and 122 essential hypertensive subjects; of the essential hypertensive subjects, 65 patients underwent non-invasive ambulatory blood pressure. There was no significant difference of ACE genotype and allele frequencies between the hypertensive and normotensive subjects. Non-significant differences were demonstrated in gender, age, causal blood pressure according to the ACE gene genotypes in the 65 patients who underwent non-invasive ambulatory blood pressure; however, effects of the ACE gene genotypes were observed on the derivatives of ambulatory blood pressure, including sleeping blood pressure levels and circadian blood pressure rhythm (p < 0.05). We suggest that insertion/deletion polymorphism of the ACE gene should be associated with blood pressure and ambulatory blood pressure might be superior to causal blood pressure at the molecular level.

Aged↗