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Biomedical subjects

H Xing

Publications and source records attributed to H Xing.

36 records · Page 2Linked to original sources
PubMed2000

Hydroxy-dibenzo crown ether stationary phase for capillary gas chromatography using sol-gel technology.

A new kind of crown ether, OH-dibenzo-14-crown-4 (OH-DB14C4), is prepared and coated onto the fused silica capillary by sol-gel process. Chromatographic characteristics including column efficiency (> 3,000 plates/m), thermal stability (to 330 degrees C) and ability of deactivation are studied. The selectivity of new stationary phase is superior to sol-gel OH-terminal silicone oil (OH-TSO) for positional isomers of some aromatic compounds such as xylene, dichlorobenzene, nitrotoluene, nitrochlorobenzene. The new stationary phase has high sample capacity for separation of small molecular mass compounds: low-molecular-mass alcohols, ethers and ketones, short-chain fatty acids and volatile amines.

Chromatography, Gas↗
PubMed1999

Nuclear localization of protein kinase U-alpha is regulated by 14-3-3.

14-3-3 proteins are intracellular, dimeric molecules that bind to and modify the activity of several signaling proteins. We used human 14-3-3zeta as a bait in the yeast two-hybrid system to screen a murine embryonic cDNA library. One interacting clone was found to encode the carboxyl terminus of a putative protein kinase. The coding sequence of the human form (protein kinase Ualpha, PKUalpha) of this protein kinase was found in GenBank(TM) on the basis of sequence homology. The two-hybrid clone was also highly homologous to TOUSLED, an Arabidopsis thaliana protein kinase that is required for normal flower and leaf development. PKUalpha has been found by coimmunoprecipitation to bind to 14-3-3zeta in vivo. Our confocal laser immunofluorescence microscopic experiments revealed that PKUalpha colocalizes with the cytoplasmic intermediate filament system of cultured fibroblasts in the G(1) phase of the cell cycle. PKUalpha is found in the perinuclear area of S phase cells and in the nucleus of late G(2) cells. Transfection of cells with a dominant negative form of 14-3-3eta promotes the nuclear localization of PKUalpha. These results suggest that the subcellular localization of PKUalpha is regulated, at least in part, by its association with 14-3-3.

14-3-3 Proteins↗
PubMed1999

[Identification and characterization of HIV-1 subtype D strain in China].

OBJECTIVE: Through sequence analysis of HIV-1 gag, tat and env regions, the subtype D HIV-1 had been identified appering in China. METHODS: The blood was collected from HIV-1 infected individual (SC9 712) in Sichuan Province. The peripheral blod mononuclear cells (PBMCs) were isolated from the blood by ficoll and the genomic DNA was extracted. The gag gene, the C2-V3 of the env gene and the first exon of tat gene were amplified by nested-PCR and sequenced directly. RESULTS: The genetic distance respectively is 7.56% in tat gene between SC9 712 and UG274A, 9.59% and 9.94% in gag gene between SC9 712 and K31 and VI203, 13.86% in env gene between SC9 712 and MAL. On the phylogenetic tree the SC9 712 HIV-1 strain clustered with the Africa subtype D strains UG274A, K31, VI203 and MAL. The above data demonstrated that SC9 712 was subtype D HIV-1 strain. Epidemiological data showed that SC9 712 virus was from a person who had worked in Africa. CONCLUSION: The results indicated that the person who came back from Africa had transmitted the subtype D of HIV-1 from Africa to China.

Base Sequence↗
PubMed1999

KSR-1 binds to G-protein betagamma subunits and inhibits beta gamma-induced mitogen-activated protein kinase activation.

The protein kinase KSR-1 is a recently identified participant in the Ras signaling pathway. The subcellular localization of KSR-1 is variable. In serum-deprived cultured cells, KSR-1 is primarily found in the cytoplasm; in serum-stimulated cells, a significant portion of KSR-1 is found at the plasma membrane. To identify the mechanism that mediates KSR-1 translocation, we performed a yeast two-hybrid screen. Three clones that interacted with KSR-1 were found to encode the full-length gamma10 subunit of heterotrimeric G-proteins. KSR-1 also interacted with gamma2 and gamma3 in a two-hybrid assay. Deletion analysis demonstrated that the isolated CA3 domain of KSR-1, which contains a cysteine-rich zinc finger-like domain, interacted with gamma subunits. Coimmunoprecipitation experiments demonstrated that KSR-1 bound to beta1 gamma3 subunits when all three were transfected into cultured cells. Lysophosphatidic acid treatment of cells induced KSR-1 translocation to the plasma membrane from the cytoplasm that was blocked by administration of pertussis toxin but not by dominant-negative Ras. Finally, transfection of wild-type KSR-1 inhibited beta1 gamma3-induced mitogen-activated protein kinase activation in cultured cells. These results demonstrate that KSR-1 translocation to the plasma membrane is mediated, at least in part, by an interaction with beta gamma and that this interaction may modulate mitogen-activated protein kinase signaling.

Animals↗
PubMed1999

Multiple docking sites on substrate proteins form a modular system that mediates recognition by ERK MAP kinase.

MAP kinases phosphorylate specific groups of substrate proteins. Here we show that the amino acid sequence FXFP is an evolutionarily conserved docking site that mediates ERK MAP kinase binding to substrates in multiple protein families. FXFP and the D box, a different docking site, form a modular recognition system, as they can function independently or in combination. FXFP is specific for ERK, whereas the D box mediates binding to ERK and JNK MAP kinase, suggesting that the partially overlapping substrate specificities of ERK and JNK result from recognition of shared and unique docking sites. These findings enabled us to predict new ERK substrates and design peptide inhibitors of ERK that functioned in vitro and in vivo.

3T3 Cells↗
PubMed1998

[Expression of mdr-1 gene in cancer tissue and its association with morphological indexes of esophageal carcinoma].

OBJECTIVE: To detect the levels of mdr-1 gene expression in fresh untreated esophageal carcinomas, and to correlate these levels to current prognostic indicators of morphology. METHODS: mdr-1 gene expression of 46 untreated esophageal carcinoma was investigated with reverse transcription polymerase chain reaction (RT-PCR), and compared with the positive incidences among differentiated grades, TNM stages and macroscopic types of cancer. RESULTS: All 46 samples were pathologically squamous cell carcinoma. The positive incidence of mdr-1 gene expression was 37% (17/46) in the whole group, and 35% (6/17), 40% (8/20), 33% (3/9) for differentiated grade I, II and III respectively. The expression rate of 33% (6/18), 40% (5/12), and 37% (6/16) was for TNM stage IIa, IIb, and III respectively. Macroscopically, the positive incidence was 37% (3/8) in constrictive, 33% (5/15) in fungating, 40% (6/14) in medullary, and 33% (3/9) in ulcerative type. There were no significant differences in each category system of morphology. CONCLUSION: Because of highly expressive level of mdr-1 gene in untreated esophageal carcinoma, we should choose appropriate chemotherapeutic regimen for esophageal Ca. The expression of mdr-1 gene in untreated esophageal cancer was independent of morphologic prognostic indexes without correlation between mdr-1 gene expression and morphological indexes.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed1997

The protein kinase KSR interacts with 14-3-3 protein and Raf.

BACKGROUND: KSR (kinase suppressor of Ras) is a recently identified putative protein kinase that positively mediates the Ras signaling pathway in the invertebrates Caenorhabditis elegans and Drosophila melanogaster. The function of vertebrate KSR is not well characterized biochemically or biologically. RESULTS: We examined the physiological role of KSR in vertebrate signal transduction using Xenopus laevis oocytes. Overexpression of KSR, in combination with overexpression of the intracellular dimeric protein 14-3-3, induced Xenopus oocyte meiotic maturation and cdc2 kinase activation; the effect of KSR and 14-3-3 on oocyte maturation was blocked by co-expression of dominant-negative Raf-1. We noted that KSR contains multiple potential binding sites for 14-3-3, and we used the yeast two-hybrid system and co-immunoprecipitation experiments to show that KSR can bind to 14-3-3. Furthermore, we demonstrated that KSR can form a complex with Raf kinase both in vitro and in cultured cells. Cell fractionation studies revealed that KSR formed a complex with 14-3-3 in both the membrane and cytoplasmic fractions of cell lysates; however, KSR only formed a complex with Raf-1 in the membrane fraction. CONCLUSIONS: Our finding suggest that KSR, 14-3-3 and Raf form an oligomeric signaling complex and that KSR positively regulates the Ras signaling pathway in vertebrate organisms.

14-3-3 Proteins↗
PubMed1997

TNF alpha-mediated inhibition and reversal of adipocyte differentiation is accompanied by suppressed expression of PPARgamma without effects on Pref-1 expression.

Tumor necrosis factor alpha (TNF alpha) is a polypeptide hormone with pleiotropic effects on cellular proliferation and differentiation. To investigate how TNF alpha inhibits and reverses adipocyte differentiation, we studied the expression of two factors involved in the adipocyte differentiation process. Peroxisome proliferator-activated receptor gamma (PPARgamma) is a positive regulator of adipogenesis, whereas preadipocyte factor 1 (Pref-1) inhibits adipocyte differentiation. The expression patterns of both PPARgamma and Pref-1 change during early stages of adipocyte differentiation. Decreased expression of Pref-1 and increased expression of PPARgamma occur 1 day and 2 days, respectively, after 3T3-L1 cells reach confluence. During TNF alpha-mediated inhibition of adipocyte differentiation, PPARgamma messenger RNA (mRNA) expression stays at low levels. In contrast, TNF alpha treatment has no effect on the normal decrease in Pref-1 gene expression that occurs during adipogenesis. We observed that certain cytokine and growth factors [such as TNF alpha, basic fibroblast growth factor, transforming growth factor beta, and protein kinase C-activating agents plus calcium ionophore], when added to differentiated adipocytes, cause rapid down-regulation of PPARgamma mRNA expression with concomitant decrease in adipocyte-specific gene expression but fail to increase Pref-1 mRNA expression. Moreover, addition of TNF alpha to fully differentiated adipocytes results in the rapid disappearance of PPARgamma protein expression and the rapid loss of PPARgamma DNA-binding activity. Therefore, Pref-1 seems to function as a nonreversible molecular checkpoint whose expression is insensitive to TNF alpha-generated signals, whereas PPARgamma expression remains sensitive to TNF alpha at all stages of the adipogenesis program. Our results support the notion that dedifferentiated adipocytes and preadipocytes are not identical, though they share many similar morphological and gene expression patterns.

3T3 Cells↗
PubMed1997

[On the effect of Chinese lacquer upon the cell division of root tip of Allium cepa].

Chinese Lacquer, as a fine coating, has been studied and applied for thousands years. The allergic reaction in Chinese Lacquer on the human skin has also been known early. The reaction of Chinese Lacquer on mitosis of cell in plant meristem have not been reported yet and was carefully studied in this paper. The result showed that Chinese Lacquer induced severe abnormality of mitotic division in Allium cepa root tips. This was more obvious in the anaphase and telophase, especially in the former phase laggard chromosomes, chromosome bridges, acentric fragments and polypolar distribution could be seen frequently. A lot of polynuclear bodies were observed in the telophase. Therefore, we think that the Chinese Lacquer can be used as a plant cell mutagen, and suggest geneticists and physiologists to do more researches on the effects of Chinese Lacquer at the genetic variation, metabolism etc.

Allium↗
PubMed1996

A molecular pathologic study on apoptosis in retinoblastoma and the mechanism of spontaneous regression in retinoblastoma.

OBJECTIVE: The present study was designed to prove the existence of apoptosis in retinoblastoma (Rb) and to determine the pathogenic mechanism of spontaneous regression of Rb as well as the relationship between them. METHODS: Qualitative morphological study on Rb was performed by means of light microscope, electron microscope and TdT mediated biotin-dUTP nick-end labeling (TUNEL). Quantitative study was performed by automatic image analysis technology (AIAT) stained with Feulgen reaction. RESULTS: The characteristic regressed area occurred in all 47 cases of Rb. Morphological changes observed within Rb closely resembled the apoptotic cell described by Kerr et al in 1972. Under electron microscope, details of apoptosis were observed in 7 cases of Rb: the morphological sequence of events occurred in and around the cell nucleus. The morphology of the TUNEL labeling Rb cell was various. TUNEL labeling showed more positive cells in regressed area, while fewer in advanced area. AIAT revealed that apoptosis index (AI) in regressed areas was higher than that in other areas of Rb, DNA average ploidy (DP) consisted with the histology grade of Rb, the degree of hyperdiploid (DH) in metastatic area was distinctly higher than that in other areas (P < 0.05). CONCLUSIONS 1. Morphologic evidence proved the existence of apoptosis, especially more in regressed area of Rb. 2. Apoptosis may participate in the spontaneous regression of Rb. Apoptosis contributed to the spontaneous regression of Rb. 3. Tumor growth parameters of Rb (AI, DP, DH) obtained by AIAT may be used as quantitative index for pathologic classification, the selection of clinical treatment and the prognostic evaluation.

Apoptosis↗
PubMed1995

An N-terminal deletion mutant of estrogen receptor exhibits increased synergism with upstream activators and enhanced binding to the estrogen response element.

To study the role of the N-terminal region of the estrogen receptor (ER) in transcription activation and in DNA binding, we constructed a mutant of the Xenopus laevis ER which lacks amino acids 1-159 (XER160/586). In transient transfections, XER160/586 exhibited < 10% of the activity of wild-type XER on a synthetic promoter containing two estrogen response elements (EREs). To examine transcriptional synergism by XER and by XER160/586, we determined the activity of promoters containing EREs and binding sites for either the vitellogenin activator, NF1, or AP1 upstream activator protein. For the three promoters transcription by XER was 2.8-fold greater than expected for additive activities, and transcription by XER160/586 was 6.2-fold greater. These data demonstrate that an upstream activator protein bound near the promoter can partially compensate for the loss of the internal N-terminal (AF1) transactivation domain in XER160/586. Using a promoter interference assay to study the intracellular interaction between ER and the estrogen response element, we found that XER160/586 exhibited a significant increase in affinity for the ERE. Its low basal activity and enhanced affinity for the ERE make XER160/586 an effective dominant negative mutant. When co-expressed with wild-type XER at 1:1 and 5:1 ratios, XER160/586 suppressed the activity of wild-type XER by 57% and > 80%, respectively.

Animals↗
PubMed1994

Antiestrogens activate an estrogen receptor mutant exhibiting enhanced binding to the estrogen response element.

The appearance of constitutively active and antiestrogen resistant estrogen receptor variants has been proposed as one of several factors leading to the development of antiestrogen resistant breast cancers. We recently described the ER2, estrogen receptor mutant, which exhibits enhanced binding to the estrogen response element on DNA, and partial constitutive estrogen-independent) ability to activate transcription. In this work we used transient transfections to show that the antiestrogens trans hydroxytamoxifen and ICI 164,384 are unable to suppress the constitutive activity of the ER2 mutant. Instead, both antiestrogens were concentration-dependent activators of the ER2 mutant. The ER2 mutant appears to be the first estrogen receptor mutant to show activation of a simple estrogen response element-containing promoter using the "pure" antiestrogen ICI 164,384.

Animals↗
PubMed1994

Characterization of gamma-glutamyltranspeptidase in the liver of the frog: 1. Comparison to the rat liver enzyme.

The characteristics of the enzyme gamma-glutamyltranspeptidase were determined in frog liver and compared to those of the rat. In Rana pipiens, tissue distribution studies indicated the order of activity to be: kidney >>> liver >> nerve > egg > lung > heart > skeletal muscle in homogenates. In the Rana pipiens relative to the Fischer 344 rat, the activity of the liver enzyme was somewhat greater (1.8-fold) and the kidney enzyme substantially less (25-fold). Frog liver gamma-glutamyltranspeptidase displayed strain-dependent differences in activity with Rana pipiens and Rana sylvatica exhibiting comparable activities and Xenopus laevis exhibiting 20-fold lower activities. No influence of sex was apparent in Rana pipiens in contrast to the sex dependent differences observed in the Fischer 344 rat: female:male = 7:1. In homogenates and plasma membrane fractions of Rana pipiens, Xenopus laevis and the Fischer 344 rat, high, and comparable relative specific activities, were observed, 8-11, coupled with protein yields of 2.2-2.5 per cent indicating the enzyme to be plasma membrane bound and associated with the sinusoidal surface of the liver cell.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Characterization of gamma-glutamyltranspeptidase in the liver of the frog: 2. Response to season, temperature and thyroid hormone in Rana pipiens.

The impact of season and temperature on frog liver gamma-glutamyltranspeptidase was assessed by measuring the activity of this enzyme in plasma membranes isolated from the livers of Rana pipiens obtained as summer and winter frogs; subjected to short-term (3 weeks) temperature acclimation; and subjected to multiple-temperature shifts. Plasma levels of T3 were determined. gamma-Glutamyltranspeptidase was found to be 2.2-fold higher in the summer frog relative to the winter frog; decreased by 44 percent in the summer frog by cold acclimation and increased by 1.7-fold in the winter frog by warm acclimation; and increased by 1.9-fold in the summer frog and 2.8-fold in the winter frog subjected to multiple-temperature shifts. Plasma T3 levels were found to be 42-fold higher in the summer frog relative to the winter frog; decreased by 42 percent by cold acclimation and increased by 2.9-fold by warm acclimation; and decreased by 39 percent and 38 percent in the summer and winter frogs subjected to multiple temperature shifts. T3 replacement during the last phase of the multiple-temperature shift protocol, restored the plasma T3 levels to 75 percent of the control levels and prevented the increase evoked by the multiple-temperature shifts in gamma-glutamyltranspeptidase activity. Indeed, enzyme activity in the T3 replaced state was 19 percent lower than in the control state. The involvement of thyroid hormone as a negative regulator of enzyme activity is discussed.

Animals↗
PubMed1993

An estrogen receptor mutant exhibiting hormone-independent transactivation and enhanced affinity for the estrogen response element.

To study transactivation by the Xenopus laevis estrogen receptor (XER), we inserted one or two copies of a synthetic amphipathic helix at amino acid 276 of the XER. The XER mutants containing one or two copies of the amphipathic helix (XER/1AH and XER/2AH, respectively) and wild-type XER were expressed at similar levels. In transient transfection assays, XER/1AH exhibited only a modest, promoter-specific increase in transactivation. Constitutive (estrogen-independent) transcription of a synthetic promoter containing two estrogen response elements (EREs) was approximately 10-fold higher for the XER/2AH mutant than for wild-type XER. The XER/2AH mutant and wild-type XER exhibited similar 17 beta-estradiol dose-response curves for transactivation. In studies carried out over a broad range of DNA concentrations using the simple 2ERE-TATA promoter or a complex vitellogenin-derived promoter, the XER/2AH mutant exhibited an estrogen-dependent 2-3-fold increase in transactivation. A 2-3-fold increase in transactivation by XER/2AH was also observed using synthetic promoters in which the two EREs exhibit synergistic interactions with the NF1, AP1, or vitellogenin activator upstream activator sequences. Using a promoter interference assay to investigate intracellular interactions between the estrogen receptor and the ERE, we showed that binding of wild-type XER to the ERE was strongly estrogen-dependent. In the presence of 17 beta-estradiol, XER/2AH and wild-type XER exhibited similar promoter interference curves. In the absence of 17 beta-estradiol, the expression plasmid encoding the XER/2AH mutant achieved levels of promoter interference with 0.25-0.5 microgram of transfected DNA that were similar to those observed with 5-10 micrograms of the expression plasmid encoding wild-type XER. The ability of the XER/2AH mutant to activate transcription in the absence of estrogen therefore is likely to be related to the approximately 20-fold increase in its apparent ability to bind to the ERE. Since XER/2AH was unable to activate transcription from a glucocorticoid response element, enhanced binding of XER/2AH to the ERE did not result from a general increase in binding to DNA. The XER/2AH mutant appears to be the first nuclear receptor mutant to retain hormone-dependent transactivation and to exhibit enhanced hormone-independent binding to its hormone response element.

Animals↗
PubMedDate not supplied

CYP2C9 Ile359Leu polymorphism, plasma irbesartan concentration and acute blood pressure reductions in response to irbesartan treatment in Chinese hypertensive patients.

Our previous study demonstrated that the CYP2C9*3 gene variant was significantly associated with elevated plasma irbesartan concentration and blood pressure decline, assessed by a 4-week follow-up and revisit following daily administration of irbesartan. We conducted a further analysis to examine the acute effects of the CYP2C9 polymorphism on plasma concentration and blood pressure through remeasurement 6 h after administration of irbesartan. We used an extreme-sampling approach by selecting individuals from the top and bottom deciles of blood pressure response residuals to irbesartan from the previous study population in Anhui, Taihu, and Dongzhi Counties, in China. A total of 196 subjects were available for the analysis. Pre- and posttreatment systolic and diastolic blood pressures (SBP and DBP), and venous blood samples (0.5, 2, and 6 h following the first treatment) were collected from each individual. Plasma irbesartan concentrations were determined by a standard HPLC/fluorescence method. The observed frequencies were 97.7% for CYP2C9*1 (Ile359) and 2.3% for CYP2C9*3 (Leu359). Subjects with the CYP2C9*1/CYP2C9*3 genotype had significantly higher plasma irbesartan concentrations when compared with those with the CYP2C9*1/CYP2C9*1 genotype (beta +/- SE = 81 +/- 36) and greater DBP response (beta +/- SE = 5.6 +/- 2.5 mmHg) at the 6-h time point after adjusting for important confounders. Our finding suggests that the CYP2C9*3 gene variant significantly alters the plasma concentration and acute DBP response at the 6-h point following irbesartan treatment in Chinese hypertensive patients.

Administration, Oral↗
PubMedDate not supplied

Characterization and evaluation of a novel exalting round-plate photocatalytic reactor.

A novel reactor, the exalting round-plate photocatalytic reactor (ERPPR), was designed and evaluated. Residence time and light intensity distributions were determined by stimulus-response experimentation and an irradiation meter, respectively. The effects of hydraulic loading of the round plate and aeration on the photocatalytic degradation of azo dye were investigated during different conditions. Experimental results show that the ERPPR is intermediate between a completely stirred tank reactor and a plug-flow reactor. Mean hydraulic loadings at different rotational speeds were significantly influenced by the reaction rate for dye decolorization. Photodegradation of the dye, however, was not significantly influenced by the amount of oxygen gas aerated, except when aerating with pure oxygen. Results further indicate that the photoreaction atmosphere of the ERPPR depends on mass transfer between gas and liquid on the round plate. The primary role of aeration is to mix catalyst and solution.

Catalysis↗