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Biomedical subjects

X F Wang

Publications and source records attributed to X F Wang.

At least 37 records · Page 2Linked to original sources
PubMed2001

A new noninvasive method for evaluation of coronary endothelial function in hypertensive patients based on change in diameter of the left main coronary artery induced by cold pressor test using echocardiography.

BACKGROUND: Coronary endothelial function is frequently studied by measuring the vasodilator response of coronary arteries to acetylcholine or to cold pressor test by invasive quantitative coronary angiography. Because invasive methods have substantial inherent limitations, studies should attempt to evaluate coronary endothelial function noninvasively. HYPOTHESIS: We attempted to evaluate the accuracy of measurement of the percent change in diameter of the left main trunk induced by cold pressor test with two-dimensional (2-D) echocardiography. Furthermore, we applied this method to the evaluation of coronary artery endothelial function in hypertensive patients. METHODS: We measured the left main trunk diameter in 21 subjects (51 +/- 4 years) before and after cold pressor test using quantitative coronary angiography followed immediately by 2-D echocardiography. The accuracy of measurement of the left main trunk diameter and its percent change by echocardiography was evaluated by comparing the values obtained by the two methods. In addition, using echocardiography, we compared left main trunk diameter responses to cold pressor test in 16 hypertensive patients [51 +/- 5 years (mean +/- standard deviation)] and 16 matched healthy subjects (50 +/- 4 years). RESULTS: Although there was only a weak correlation between the absolute values of the left main trunk diameter measured by the two methods (r = 0.61; p = 0.04), a strong correlation was found between the percent change in diameter measured by the two methods (r = 0.93; p = 0.0001). The percent change in diameter of the left main trunk induced by cold pressor test in hypertensive patients (-3.7 +/- 10.6%) was significantly lower than that in control subjects (13.2 +/- 6.8%, p = 0.0001). CONCLUSIONS: Percent change in diameter of the left main trunk induced by cold pressor test can be evaluated accurately using 2-D echocardiography. Our study showed reduced vasodilation or vasoconstriction of the left main trunk after cold pressor test in hypertensive patients compared with healthy subjects, indicating impaired coronary endothelial function in hypertensive patients. The present echocardiographic method is a potentially useful new noninvasive method for evaluating coronary endothelial function.

Adult↗
PubMed2001

Suppression of ATP-induced Cl(-)secretion by enhanced expression of epithelial Na(+)channels in mouse endometrial epithelium.

We have studied the effect of enhanced expression of epithelial Na(+)channels (ENaC) on the ATP-induced Cl(-)secretion in the mouse epithelium using short-circuit current (I(SC)) and RT-PCR techniques. The amiloride sensitivity of basal current (I(b)) across the cultured endometrial epithelia was found to vary with the magnitude of the I(b), the higher the I(b)the greater its sensitivity to amiloride, indicating possible elevation of ENaC. However, the magnitude of ATP-induced I(SC), previously demonstrated to be mediated by Ca(2+)-activated chloride channel (CaCC), decreased as the amiloride sensitivity of the I(b)increased, suggesting a possible inhibitory effect of elevated expression of ENaC on ATP-mediated chloride secretion. The Matrigel treatment for culturing the endometrial epithelia affected the amiloride sensitivity of the I(b)as well as the ATP-induced I(SC)reversedly. Competitive RT-PCR demonstrated that the expression of both ENaC gamma subunits and CaCC was enhanced in Matrigel-treated cultures. However, the observed reduction in the ATP-induced or CaCC-mediated I(SC)could not be explained by the CaCC expression pattern. These data suggest that inhibition of CaCC function is due to enhanced ENaC expression. Therefore, in addition to interacting with CFTR, ENaC also appears to interact with CaCC in the mouse endometrial epithelium. Physiologically the present findings indicate that enhanced expression of ENaC leads to suppression of other Cl(-)channels, such as CFTR and CaCC, thereby preconditioning the endometrium in favour of overall salt and water absorption as observed during embryo implantation.

Adenosine Triphosphate↗
PubMed2001

The role of Smad3 in mediating mouse hepatic stellate cell activation.

Transforming growth factor beta (TGF-beta) is the most potent profibrogenic mediator in liver fibrosis. Although Smad proteins have been identified as intracellular mediators in the TGF-beta signaling pathway, the function of individual Smad proteins remains poorly understood. The aim of this study was to explore the contribution of Smad3 in mediating TGF-beta responses in a model of acute liver injury in vivo and in culture-activated hepatic stellate cells (HSCs). Wild-type, Smad3 heterozygous or Smad3 homozygous knockout mice were treated with a single intragastric administration of CCl(4). After 72 hours, the induction of hepatic collagen alpha1(I) and alpha2(I) messenger RNA (mRNA) levels in Smad3 knockout mice was only 42% and 64%, respectively, of the levels induced in wild-type mice. However, smooth muscle alpha-actin (alpha-SMA) was expressed at a slightly higher level in livers from knockout mice compared with wild-type mice. In culture-activated HSCs from Smad3 knockout mice, collagen alpha1(I) mRNA was 73% of wild-type HSCs, but alpha-SMA expression was the same. HSCs from knockout mice showed a higher proliferation rate than wild-type HSCs. Smad3-deficient HSCs did not form TGF-beta1-induced Smad-containing DNA-binding complexes. In conclusion, (1) maximal expression of collagen type I in activated HSCs requires Smad3 in vivo and in culture; (2) Smad3 is not necessary for HSC activation as assessed by alpha-SMA expression; (3) Smad3 is necessary for inhibition of proliferation of HSCs, which might be TGF-beta-dependent; and (4) Smad3 is required for TGF-beta1-mediated Smad-containing DNA-binding complex formation in cultured HSCs.

Actins↗
PubMed2001

The loss of Smad3 results in a lower rate of bone formation and osteopenia through dysregulation of osteoblast differentiation and apoptosis.

Smad3 is a well-characterized intracellular effector of the transforming growth factor beta (TGF-beta) signaling pathway and was implicated recently in the potentiation of vitamin D receptor (VDR)-mediated signaling. Given that both TGF-beta and vitamin D are important regulators of bone remodeling, it is expected that Smad3 plays an integral role in normal maintenance of bone. However, the exact mechanisms by which Smad3 functions in bone remodeling are unknown. Here, we show that mice with targeted deletion of Smad3 are osteopenic with less cortical and cancellous bone compared with wild-type littermates. Decreases in bone mineral density (BMD) in Smad3 null mice reflect the inability of osteoblasts to balance osteoclast activity, although osteoclast numbers are normal and vitamin D mediated serum calcium homeostasis is maintained. The osteopenia of Smad3 null mice is attributed to a decreased rate of bone formation associated with increased osteocyte number and apoptosis. These findings are supported by studies with isolated primary osteoblasts that show TGF-beta can no longer inhibit the differentiation of osteoblasts in the absence of Smad3; yet, TGF-beta-stimulated proliferation remains intact. Together these data support a model that a loss of Smad3 increases the osteocyte fate of the osteoblast and decreases the duration of osteoblast function by shortening lifespan, ultimately resulting in osteopenia.

Animals↗
PubMed2001

Enhanced epithelial Na(+) channel (ENaC) activity in mouse endometrial epithelium by upregulation of gammaENaC subunit.

The amiloride-sensitive epithelial Na(+) channel (ENaC), which is made of three different but homologous subunits, controls the rate of transepithelial Na(+) absorption in a variety of epithelia. The present study investigated the functional role of its subunits in regulating ENaC activity, measured as amiloride sensitive short-circuit current (I(SC)), in the mouse endometrial epithelium under different culture conditions. The treatment of the cultured epithelia with aldosterone (1 microM) or culturing cells on filters coated with concentrated Matrigel resulted in an increase in the amiloride-sensitive I(SC). Semiquantitative RT-PCR demonstrated that the expression of alpha and beta subunits was not significantly altered by these treatments, but an increase in the gamma subunit expression was observed. An 11-fold increase, induced by aldosterone, in the expression of the gamma subunit, but not in the alpha and beta subunits, was confirmed by capillary electrophoresis with laser-induced fluorescence (CE-LIF). The treatment of endometrial cells with antisense against the gammaENaC subunit abolished the aldosterone-enhanced amiloride-sensitive I(SC). The results indicated an important role of gammaENaC subunit in determining ENaC activity, and a possible role of the gammaENaC subunit in interacting with CFTR was also discussed.

Absorption↗
PubMed2001

[Comparison of immune responses elicited by recombinant protein and eukaryotic expression plasmid based on histidine rich protein 2 of Plasmodium falciparum].

OBJECTIVE: To identify the immune characteristics of different vaccine prototypes based on HRP2 and to provide experimental evidence for developing P. f. blood stage vaccines. METHODS: BALB/c mice were immunized with recombinant protein TP-HRP2 or eukaryotic expression plasmid pcDNA3.1(-)/HRP2. The kinetics and specificities of antibody responses were analyzed. The proliferation tests of spleen cells were done, and P. f. growth inhibition assays were done with immune sera. RESULTS: The mice immunized with TP-HRP2 in Freund's adjuvant produced high-level and high-specificity antibody response. The antibodies appeared rapidly and lasted for a longer time. Cellular responses were induced simultaneously, and the immune sera could inhibit the development of parasite in IRBCs. The mice immunized with pcDNA3.1(-)/HRP2 produced middle-level antibody response which had some specificity, however, the induction of antibodies required repeated inoculation and a longer duration. Immune cells were well primed and the memorial immune response was obvious but the immune sera had no effect on the growth of P.f. in vitro. CONCLUSION: Both the recombinant protein and plasmid DNA based on HRP2 have different immune characteristics in mice. HRP2 recombinant protein has the potential in practical application.

Animals↗
PubMed2001

[Study on the characters of hydrogen bonds in protein and nucleic acid of the breast cancer tissues].

Some remarkable spectral differences are observed between the normal breast tissues and breast cancer tissues, including those of association pattern and degree of the hydrogen bonds in the protein and nucleic acid biomacromolecules. These differences are found in the relative intensity, absorption position and shape of the characteristic bands: (1) amide I band, stretching vibration bands of N-H group and C-O(H) in the residues of some amino-acids, and (2) the relative intensity of asymmetric stretching vibration of the phosphodiester group in nucleic acid. The hydrogen bond is the major force which maintain and promote the formation of the high structure of protein and nucleic acid. It is possible to diagnose the breast cancer and forecast the possibility of canceration using above spectral characters of the hydrogen bond with other spectral characters.

Amides↗
PubMed2000

Suppression of CFTR-mediated Cl(-) secretion by enhanced expression of epithelial Na(+) channels in mouse endometrial epithelium.

The present study investigated the effect of enhanced expression of epithelial Na(+) channels (ENaC) on the cystic fibrosis transmembrane conductance regulator (CFTR)-mediated Cl(-) secretion in the mouse endometrium using the short-circuit current technique. The amiloride sensitivity of the basal current of the cultured endometrial epithelia was found to vary with the magnitude of the basal current, the higher the basal current the greater its sensitivity to amiloride, indicating possible elevation of ENaC expression. However, the magnitude of the forskolin-induced Isc, previously demonstrated to be mediated by CFTR, decreased as the amiloride sensitivity of the basal current increased, suggesting a possible inhibitory effect of elevated expression of ENaC on CFTR-mediated Cl(-) secretion. The Matrigel concentration for culturing the endometrial epithelia was found to affect the amiloride sensitivity of the basal current as well as the forskolin-induced Isc in opposite directions. However, competitive RT-PCR demonstrated that the expression of both ENaC and CFTR was enhanced in Matrigel-treated culture, suggesting that the reduced forskolin-induced Isc with enhanced amiloride sensitivity was not due to a reduction in CFTR expression, but rather suppression of CFTR function by enhanced ENaC expression. In addition to the previously demonstrated inhibition of ENaC by activation of CFTR, the present results reveal possible regulation of CFTR by ENaC. The interaction between the two may be one of the underlying mechanisms for balancing Na(+) absorption and Cl(-) secretion across epithelia.

Animals↗
PubMed2000

Pyrimidinoceptors-mediated activation of Ca(2+)-dependent Cl(-) conductance in mouse endometrial epithelial cells.

Previous studies have demonstrated the activation of endometrial Cl(-) secretion through P(2Y2) (P(2U)) purinoceptors by extracellular ATP. The present study further explored the presence of pyrimidine-sensitive receptors in the primary cultured mouse endometrial epithelial cells using the short-circuit current (I(SC)) and whole-cell patch-clamp techniques. UDP induced a transient increase in I(SC) in a concentration-dependent manner (EC(50) approximately 8.84 microM). The UDP-induced I(SC) was abolished after pretreating the epithelia with a calcium chelator, 1, 2-bis-(2-aminophenoxy)-ethane-N,N,N'N'tetraacetic acid-acetomethyl ester (BAPTA-AM), suggesting the dependence of the I(SC) on cytosolic free Ca(2+). The type of receptor involved was studied by cross-desensitization between ATP and UDP. ATP or UDP desensitized its subsequent I(SC) response. However, when ATP was added after UDP, or vice versa, a second I(SC) response was observed, indicating the activation of distinct receptors, possibly pyrimidine-sensitive receptors in addition to P(2Y2) (P(2U)) receptors. Similar results were observed in the patch-clamp experiments where UDP and ATP were shown to sequentially activate whole-cell current in the same cell. The UDP-activated whole-cell current exhibited outward rectification with delay activation and inactivation at depolarizing and hyperpolarizing voltages, respectively. In addition, the UDP-evoked whole-cell current reversed near the equilibrium potential of Cl(-) in the presence of a Cl(-) gradient across the membrane, and was sensitive to 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), indicating the activation of Ca(2+)-activated Cl(-) conductance. These characteristics were very similar to that of the ATP-activated whole-cell current. Taken together, our findings indicate the presence of distinct receptors, pyrimidinoceptors and P(2Y2) (P(2U)) receptors in mouse endometrial epithelial cells. These distinct receptors appear to converge on the same Ca(2+)-dependent Cl(-) channels.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2000

Modulation of the Ca(2+)-activated Cl(-) channel by 14-3-3epsilon.

We have previously reported an association of 14-3-3epsilon isoform with calmodulin. Using the voltage-clamp technique, the present study investigated the potential role of 14-3-3 in modulating the Ca(2+)-activated Cl(-) channel (CaCC) endogenously expressed in Xenopus oocytes. Injection of 14-3-3epsilon antisense oligodeoxynucleotides resulted in potentiation of the ionomycin-induced Cl(-) current, while 14-3-3 peptide and calmodulin inhibitor, W13, suppressed the antisense-potentiated current. The data suggest that 14-3-3epsilon plays an inhibitory role in modulating the CaCC by interacting with the calmodulin-dependent pathway. The potential role of 14-3-3epsilon in other tissues and its therapeutic potential for cystic fibrosis are discussed.

14-3-3 Proteins↗
PubMed2000

Acrylamide-regulated neurofilament expression in rat pheochromocytoma cells.

Using the rat pheochromocytoma cell line (PC12), we present molecular evidence that the neurotoxicant acrylamide directly induces neurofilament gene expression, and the signaling pathways are initially distinctive from, but eventually merged into, that for nerve growth factor (NGF)-induced neurofilament expression. In PC12 cells, acrylamide increased neurofilament protein levels and synthesis. Acrylamide had no effect on the stability of neurofilament mRNAs suggesting that it directly increased neurofilament mRNA synthesis. K252a, a selective inhibitor for NGF receptor gp140trk, had no effect on acrylamide induction, but completely inhibited NGF-induced neurofilament protein synthesis. Therefore, the initial step for acrylamide signaling was distinctive from NGF. Dexamethasone reversed the effects of both NGF and acrylamide on neurofilament protein levels and synthesis indicated that there is a dexamethasone-sensitive signaling step upon which NGF and acrylamide merge, suggesting involvement of transcription-activating proteins like AP-1. These results, taken together with previous studies of transgenic mice that overexpress neurofilament genes, may partially explain the mechanisms of neurofilament accumulation in distal parts of large axons, a pathognomonic feature of acrylamide neurotoxicity in animals.

Acrylamide↗
PubMed2000

Astrocytes provide cysteine to neurons by releasing glutathione.

Cysteine is the rate-limiting precursor of glutathione synthesis. Evidence suggests that astrocytes can provide cysteine and/or glutathione to neurons. However, it is still unclear how cysteine is released and what the mechanisms of cysteine maintenance by astrocytes entail. In this report, we analyzed cysteine, glutathione, and related compounds in astrocyte conditioned medium using HPLC methods. In addition to cysteine and glutathione, cysteine-glutathione disulfide was found in the conditioned medium. In cystine-free conditioned medium, however, only glutathione was detected. These results suggest that glutathione is released by astrocytes directly and that cysteine is generated from the extracellular thiol/disulfide exchange reaction of cystine and glutathione: glutathione + cystine<-->cysteine + cysteine-glutathione disulfide. Conditioned medium from neuron-enriched cultures was also assayed in the same way as astrocyte conditioned medium, and no cysteine or glutathione was detected. This shows that neurons cannot themselves provide thiols but instead rely on astrocytes. We analyzed cysteine and related compounds in rat CSF and in plasma of the carotid artery and internal jugular vein. Our results indicate that cystine is transported from blood to the CNS and that the thiol/disulfide exchange reaction occurs in the brain in vivo. Cysteine and glutathione are unstable and oxidized to their disulfide forms under aerobic conditions. Therefore, constant release of glutathione by astrocytes is essential to maintain stable levels of thiols in the CNS.

Animals↗
PubMed2000

Adenosine triphosphate induces inhibition of Na(+) absorption in mouse endometrial epithelium: a Ca(2+)-dependent mechanism.

The present study investigated the inhibitory effect of extracellular ATP on Na(+) absorption and the possible underlying mechanism in cultured mouse endometrial epithelium using the short-circuit current (I(SC)) technique. The cultured epithelia exhibited a Na(+)-dependent basal current that could be predominately blocked by the epithelial Na(+) channel (ENaC) blocker, amiloride (10 microM). Apical addition of ATP (10 microM) induced a reduction in basal I(SC). However, in the presence of amiloride or when apical Na(+) was removed, the ATP-induced reduction was abolished and an increase in the I(SC) was observed with kinetic characteristics similar to those reported previously for the ATP-induced Cl(-) secretion, indicating that ATP could induce both Cl(-) secretion and inhibition of Na(+) absorption. Further reduction in I(SC) after ATP challenge could be obtained with forskolin (10 microM), which indicates that different inhibitory mechanisms are involved. The ATP-induced inhibition of Na(+) absorption, but not that induced by forskolin, could be abolished by the P(2) receptor antagonist, reactive blue (100 microM), indicating the involvement of a P(2) receptor in mediating the ATP response. ATP and uridine 5'-diphosphate (UDP; 100 microM), a relatively selective agonist for the pyrimidinoceptor, induced separate I(SC) reduction, and distinct I(SC) increases in the presence of amiloride, regardless of the order of drug administration, indicating the involvement of two receptor populations. The ATP-induced inhibition of Na(+) absorption was mimicked by the Ca(2+) ionophore, ionomycin (1 microM), whereas the Ca(2+) chelators, EGTA and BAPTA-AM, abolished the ATP-induced, but not the forskolin-induced, inhibition of Na(+) absorption, suggesting the involvement of a Ca(2+)-dependent pathway. In the presence of the Cl(-) channel blocker, DIDS (100 microM), both inhibitory and stimulatory responses to ATP were abolished, suggesting the involvement of a Ca(2+)-activated Cl(-) channels (CaCCs) in mediating both ATP responses. The ATP-induced as well as the forskolin-induced reduction in I(SC) was not observed when Cl(-) was removed from the bathing solution, indicating that Cl(-) permeation is important for the inhibition of Na(+) absorption. The results suggest the presence of a Ca(2+)-dependent ENaC-inhibiting mechanism involving CaCC in mouse endometrial epithelial cells. Thus, extracellular nucleotides may play an important role in the fine-tuning of the uterine fluid microenvironment by regulating both Cl(-) secretion and Na(+) absorption across the endometrium.

Absorption↗
PubMed2000

[High-level expression of BYDV GAV coat protein gene in Escherichia coli].

The IPTG-inducible expression vector containing the BYDV GAV coat protein gene was constructed and transferred into E. coli BL21(DE3). High-level expression of the specific protein was achieved by IPTG induction. The results of SDS-PAGE and Western blotting show that the expression product which accumulates 19.5% of the total cellular proteins estimated by scanning is 24 kD BYDV GAV coat protein plus eleven amino acids of pET-5a.

Capsid↗
PubMed2000

[Immune response in mice induced by C terminal encoding gene of Plasmodium falciparum histidine rich protein. 2].

OBJECTIVE: To explore the humoral and cellular immune responses in mice to eukaryotic expression recombinant plasmid encoding histidine rich protein 2 (HRP-II) of Plasmodium falciparum. METHODS: The start and stop codes were introduced into HRP-II gene fragment, the reading frame and the position of start and stop codes in HRP-II were identified by sequencing. HRP-II fragment containing the start and stop codes was cloned into pcDNA3.1 (-) to form pcDNA3.1 (-)/HRP-II. The BALB/c mice were immunized i.m. with the plasmids for 3 times in 3 weeks intervals. Two weeks after the last immunization, the sera and splenocytes were collected to investigate anti-HRP-II antibodies by ELISA and the splenocytes proliferation response to HRP-II. RESULTS: Sequence data show that the reading frame and the position of start and stop codes are correct. Restriction enzyme digestion indicated that the HRP-II gene fragment containing start and stop codes was successfully cloned into pcDNA3.1 (-). Mice raised significant anti-HRP-II antibodies after pcDNA3.1 (-)/HRP-II immunization, and the splenocytes proliferated prominently when stimulated with HRP-II protein. CONCLUSION: Eukaryotic expression recombinant plasmid encoding HRP-II gene can induce significantly humoral and cellular immune response in mice. HRP-II gene may be a good candidate for P. falciparum blood-stage multiple DNA vaccine.

Animals↗