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Biomedical subjects

X Wei

Publications and source records attributed to X Wei.

At least 145 records · Page 8Linked to original sources
PubMed1996

Pathogenesis of caudal dysgenesis/sirenomelia induced by ochratoxin A in chick embryos.

Caudal dysgenesis/sirenomelia is a malformation complex for which the pathogenesis is controversial. This report describes the particular vulnerability of specific caudal structures to Ochratoxin A (OA), a fungal toxin, as the basis for caudal dysgenesis in an avian model. The experimental procedure involved injection of 1 microgram of OA into the air sac of eggs that had been incubated for 48 hours prior to treatment (i.e., embryos that had reached Hamburger and Hamilton stage 9-10 (6-10 somite pairs) [Hamburger and Hamilton (1951) Dev. Dyn. 195:231-272] by the time of treatment). Six to twelve hours following OA injection, excessive cell death, as shown by vital staining and routine histology, was evident in selected cell populations, including cells of the caudal-most mesoderm (the mesoderm that apparently forms the external genitalia and median infraumbilical region), the tail bud, and the neural tube caudal to the wing buds (corresponding to the level of the presomitic mesoderm). The notochord was not severely affected, although there were degenerative changes in the presomitic mesoderm. Except for positional abnormalities, development of the lateral plate mesoderm from which the leg buds are derived appeared relatively normal in most of the treated embryos. Six days post-treatment, varying degrees of caudal dysgenesis, presenting in severely affected specimens as sirenomelia, were observed in approximately 30% of the surviving treated embryos. The potential basis for the differential vulnerability of the affected cell populations and, therefore, the cellular basis for the genesis of caudal dysgenesis/sirenomelia in this model are discussed.

Animals↗
PubMed1996

Turnover of the cystic fibrosis transmembrane conductance regulator (CFTR): slow degradation of wild-type and delta F508 CFTR in surface membrane preparations of immortalized airway epithelial cells.

The protein product of the cystic fibrosis (CF) gene, termed the cystic fibrosis transmembrane conductance regulator (CFTR), is known to function as an apical chloride channel at the surface of airway epithelial cells. It has been proposed that CFTR has additional intracellular functions and that there is altered processing of mutant forms. In examining these functions we found a stable form of CFTR with slow turnover in surface membrane preparations from CF and non-CF immortalized airway epithelial cell lines. The methods used to study the turnover of CFTR were pulse/chase experiments utilizing saturation labeling of [35S] Met with chase periods of 5-24 h in the presence of 8 mM Met and cell fractionation techniques. Preparations of morphologically identifiable surface membranes were compared to total cell membrane preparations containing intracellular membranes. Surface membrane CFTR had lower turnover defined by pulse/chase ratios than that of the total cell membrane preparations. Moreover, mutant CFTR was stable in the surface membrane fraction with little degradation even after a 24 h chase, whereas wild-type CFTR had a higher pulse/chase ratio at 24 h. In the presence of 50 microM castanospermine, which is an inhibitor of processing alpha-glucosidases, a more rapid turnover of mutant CFTR was found in the total cell membrane preparation, whereas wild-type CFTR had a lower response. The results are compatible with a pool of CFTR in or near the surface membranes which has an altered turnover in CF and a glycosylation-dependent alteration in the processing of mutant CFTR.

Amino Acid Sequence↗
PubMed1996

The postnatal development of the insertions of the medial collateral ligament in the rat knee.

Bone soft tissue remodelling at the femoral and tibial insertions of the medial collateral ligament (MCL) of the rat knee was monitored at regular intervals from birth to 120 days of age in 40 Sprague Dawley rats. At birth the femoral insertion originated from the perichondrium of the epiphysis. By day 8 the perichondrium within the insertion had turned into fibrocartilage. Secondary ossification of the femoral epiphysis had progressed in the region near to the insertion site by day 15. The epiphyseal cartilage was entirely replaced by bone by day 40 except for the fibrocartilage within the insertion. After that stage, no qualitative change in zonal insertion characteristics was observed, but only increase in size and decrease in cellularity. At birth, the tibial ligament inserted onto the thin cortical bone of the metaphysis via periosteum. At day 8, osteoclasts started to resorb the thin cortical bone at the ligament insertion, thus forming a metaphyseal depression between days 10 and 20. From days 20 to 120, the insertion remained qualitatively unchanged, showing three zones, the ligament, periosteum, and metaphyseal trabecular bone. The deep periosteal layer showed osteoclastic activity in the proximal part and osteoblastic activity in the distal part. The migration-mechanism of the ligament insertion during growth seems to be caused by this growth-related osteoclastic resorption of the proximal metaphyseal bone and by simultaneous osteogenic activity, which successively cements the distal part of the ligament to bone. The persistence of the periosteal layer and the metaphyseal depression for up to 120 days may be regarded as a sign of continuing growth in this animal model. This is the first investigation showing that the formation of the metaphyseal depression is a purely postnatal event, and suggests that this process might be initiated by the change in mode of growth and joint biomechanics after birth, enabling ligament development and migration in a growing and increasingly loaded weight-bearing joint. The mainly resorptive process, which takes place during development of the tibial MCL insertion, may account for the tensile failure of this ligament that commonly occurs at this site during growth. The pronounced morphological differences between the chondral femoral and the periosteal tibial attachment of the adult MCL are apparently caused by the different postnatal development processes at epiphyses and metaphyses.

Animals↗
PubMed1996

Effective gating charges per channel in voltage-dependent K+ and Ca2+ channels.

In voltage-dependent ion channels, the gating of the channels is determined by the movement of the voltage sensor. This movement reflects the rearrangement of the protein in response to a voltage stimulus, and it can be thought of as a net displacement of elementary charges (e0) through the membrane (z: effective number of elementary charges). In this paper, we measured z in Shaker IR (inactivation removed) K+ channels, neuronal alpha 1E and alpha 1A, and cardiac alpha 1C Ca2+ channels using two methods: (a) limiting slope analysis of the conductance-voltage relationship and (b) variance analysis, to evaluate the number of active channels in a patch, combined with the measurement of charge movement in the same patch. We found that in Shaker IR K+ channels the two methods agreed with a z congruent to 13. This suggests that all the channels that gate can open and that all the measured charge is coupled to pore opening in a strictly sequential kinetic model. For all Ca2+ channels the limiting slope method gave consistent results regardless of the presence or type of beta subunit tested (z = 8.6). However, as seen with alpha 1E, the variance analysis gave different results depending on the beta subunit used. alpha 1E and alpha 1E beta 1a gave higher z values (z = 14.77 and z = 15.13 respectively) than alpha 1E beta 2a (z = 9.50, which is similar to the limiting slope results). Both the beta 1a and beta 2a subunits, coexpressed with alpha 1E Ca2+ channels facilitated channel opening by shifting the activation curve to more negative potentials, but only the beta 2a subunit increased the maximum open probability. The higher z using variance analysis in alpha 1E and alpha 1E beta 1a can be explained by a set of charges not coupled to pore opening. This set of charges moves in transitions leading to nulls thus not contributing to the ionic current fluctuations but eliciting gating currents. Coexpression of the beta 2a subunit would minimize the fraction of nulls leading to the correct estimation of the number of channels and z.

Animals↗
PubMed1996

Relation between plasma and saliva concentrations of enoxacin, ciprofloxacin, and theophylline.

To assess the reliability of predicting plasma concentrations of enoxacin, ciprofloxacin, and theophylline from drug concentrations in saliva, six healthy volunteers received single oral doses of enoxacin, ciprofloxacin, and theophylline administered in combination on each of four separate study days, with different, doses separated by at least 5 days. Drug concentrations were determined by a newly developed high-performance liquid chromatography (HPLC) assay, which could measure simultaneously all three drugs in plasma or saliva. Saliva data from the postabsorptive phase after drug administration were used to minimize the effects of variation in absorption. There were good correlations between saliva and plasma concentrations of enoxacin, ciprofloxacin, and theophylline (r = 0.91, 0.88, and 0.98, respectively). The mean (+/-SD) saliva-to-plasma (S/P) ratio for theophylline was 0.63 +/- 0.06 with a coefficient of variation (CV) of 7.9 +/- 2.7%. In contrast, the S/P ratios and CV values for enoxacin and ciprofloxacin were 0.72 +/- 0.21 and 28.9 +/- 11.1%, and 0.58 +/- 0.15 and 25.3 +/- 6.7%, respectively. Because of the large inter- and intraindividual variability, saliva concentrations of enoxacin and ciprofloxacin are not reliable for predicting plasma concentrations. However, saliva may be used reliably for predicting plasma concentrations of theophylline.

Adult↗
PubMed1996

Protein phosphatases independently regulate vesicle movement and microtubule subpopulations in hepatocytes.

To investigate the regulation of microtubule (MT)-based vesicle transport and the interphase MT array in hepatocytes, we have used okadaic acid (OKA) and microcystin (MCYST), two toxins that inhibit serine-threonine protein phosphatases (PP) 1 and 2A, to alter cellular phosphorylation. Video-enhanced differential interference contrast microscopy analysis revealed that both toxins inhibited the frequency, velocity, and run length of MT-dependent vesicle movements dose dependently between 50 and 500 nM. At our maximum dose of 500 nM, both toxins significantly decreased PP2A activity (OKA to 45 +/- 12% and MCYST to 57 +/- 2%), whereas PP1 was inhibited only by MCYST. Because no additional effects on vesicle movements were caused by MCYST over the changes caused by OKA, these data implicate PP2A in the regulation of MT-dependent vesicle movement. To understand whether the changes in parameters of vesicle movements were due to changes in the MT array, the effects of these toxins on MT distribution were examined by immunofluorescence microscopy. Although lower doses of OKA produced no effects, treatment with 500 nM OKA altered MT organization and also caused fragmentation and loss of acetylated (stable) MTs. In contrast, MCYST concentrations up to 500 nM elicited no changes in MT organization in general or in the acetylated (stable) array. From these findings we conclude that inhibition of MT-dependent vesicle movement by the PP inhibitors, MCYST and OKA, in hepatocytes cannot result from changes or disruption in the MT array. Because OKA (an inhibitor of PP2A only in our system) at high doses caused loss of stable MTs, whereas MCYST (an inhibitor of both PP1 and PP2A) did not, we conclude that the control of the preservation of the stable MT array in hepatocytes is complex. Stable MTs require active PP2A for maintenance, but the disruption of the array through inhibition of PP2A can be prevented if PP1 is also inhibited, suggesting that the relative degree of phosphorylation of multiple cellular components is the determinant of MT stability.

Animals↗
PubMed1996

Histopathology of enzootic ataxia in Sika deer (Cervus nippon Temminck).

Thirteen Sika deer (Cervus nippon Temminck) showing ataxia on a stock farm in the north eastern district of China were examined histopathologically. The principal pathological changes were spongy vacuolation and myelin deficiency in the white matter of the spinal cord and brain stem, fibrosis and rupture of the elastic lamina of the spinal arterioles, and mesothelial hyperplasia in the spinal arachnoid. Other findings included defective formation of the elastic laminae of the aorta, and the blood vessels in the kidney and lung, hemosiderosis in the spleen and liver, and lymphocyte depletion in the systemic lymph nodes. Copper concentrations were low in the serum and liver. In the white matter of the spinal cord and brain stem, demyelination appeared to coexist with dysmyelination and secondary myelin breakdown. It was inferred that decreased activity of copper containing enzyme induced various lesions. The possible role of copper deficiency in the pathogenesis of the ataxic conditions is discussed.

Animals↗
PubMed1996

Concentrations of proteoglycan fragments in relation to maturation, sex and time of day: physiologic variations in knee joint fluid of rabbits.

We analyzed the concentrations of proteoglycan fragments in knee joint fluid in 142 rabbits to investigate the effect of physiologic variations--i.e., maturation, sex and time of day. The concentrations of proteoglycan fragments differed significantly between young, adolescent and adult animals and showed an inverse correlation to the stage of maturation of the rabbit. Adolescent male rabbits had higher concentrations than age-matched females. Morning and evening samples had similar concentrations. No relation was found between the proteoglycan fragment concentrations in joint fluid and the cartilage mass. The proteoglycan fragment concentrations in knee joint fluid apparently reflect the metabolic status of growing articular cartilage. There are considerable physiologic variations associated with maturation and sex, and these need to be taken into account when using the proteoglycan fragment concentration as a marker for joint diseases.

Age Factors↗
PubMed1996

Subunit composition is a major determinant in high affinity binding of a Ca2+ channel blocker.

Skeletal muscle L-type channels are the pharmacological receptors for Ca2+ channel antagonists, including dihydropyridines (DHPs). High affinity DHP binding to these channels in skeletal muscle membranes has been reported to be independent of Ca2+ addition and to become dependent on Ca2+ after solubilization. The channel is a multimeric complex composed of alpha 1, beta, gamma, and alpha 2 delta, of which alpha 1 is the pore-forming and DHP-binding component. In this study we coexpressed non-alpha 1 components with alpha 1 in L and COS cells and investigated their roles in the regulation of high affinity DHP binding by Mg2+ and Ca2+. No DHP binding to membranes of cells expressing alpha 1 beta alone was detected in the absence of Ca2+ or Mg2+. Addition of Mg2+ revealed the presence of (+)-PN200-110 (DHP) binding sites with a Kd of 1 nM. This affinity was 4-fold lower than that of skeletal muscle membrane binding sites (Kd = 0.25 nM). Addition of Ca2+ increased the affinity for DHP in membranes from alpha 1 beta-expressing cells to that seen in skeletal muscle membranes (Kd = 0.2-0.3 nM; EC50 of 0.2 microM). Ca2+ did not affect DHP binding to skeletal muscle membranes. Coexpression of all of the subunits completely recapitulated the high affinity DHP binding seen with skeletal muscle membranes in the absence of Ca2+ and Mg2+ (Kd = 0.15 nM). This affinity was unaffected by addition of Ca2+ or Mg2+. Coexpression of alpha 1 beta with either alpha 2 delta or gamma alone resulted in DHP binding intermediate between levels seen with alpha 1 beta and alpha 1 beta alpha 2 delta gamma. Thus, this study demonstrates that alpha 2 delta and gamma are essential for full reconstitution of the DHP binding characteristics of the skeletal muscle L-type Ca2+ channel/DHP receptor.

Aniline Compounds↗
PubMed1996

Increase in Ca2+ channel expression by deletions at the amino terminus of the cardiac alpha 1C subunit.

The alpha 1 subunit of the cardiac L-type Ca2+ channel (alpha 1C) is one of the many alternatively spliced products of a single gene that is expressed in a number of excitable tissues. Sequence comparison indicates that the amino terminus is a site of significant structural diversity. To explore the role of the amino terminus of alpha 1C in expression and function of Ca2+ channels, we constructed a series of deletion mutants of the rabbit cardiac alpha 1C subunit and expressed them in Xenopus oocytes. Deletions of up to 120 amino acids from the amino terminus increased both ionic and gating currents by 5- to 8-fold. Ca2+ currents induced by these mutants had voltage-dependent activation, inactivation, modulation by beta subunits, and single channel conductance similar to the wild type cardiac alpha 1C (wt alpha 1C). Thus, deletion of a major portion of the amino terminus of alpha 1C did not alter the three dimensional conformation essential for channel function, but enhanced the expression of Ca2+ channels in Xenopus oocytes. A deletion mutant lacking the first 171 amino acids did not yield any measurable current.

Amino Acid Sequence↗
PubMed1996

Reconstitution of the skeletal muscle dihydropyridine receptor. Functional interaction among alpha 1, beta, gamma and alpha 2 delta subunits.

The L-type voltage-dependent Ca2+ channel purified from skeletal muscle by virtue of its dihydropyridine (DHP) binding activity, is composed of alpha 1, alpha 2 delta, beta and gamma subunits. The alpha 1 subunit has the ability to function alone as a Ca2+ channel and a receptor for DHP and other Ca2+ channel antagonists. In this study, the non-alpha 1 components coexpressed with alpha 1 in COS cells were investigated for their effects on DHP binding and suppression of an anomalous allosteric regulation of the phenylalkylamine (-)D600 in complexes lacking one or more subunits. (-)D600 increased DHP binding to membranes of COS cells expressing alpha 1 beta while it did not affect DHP binding to skeletal muscle membranes. Coexpression of gamma or alpha 2 delta with alpha 1 beta partially suppressed this effect. Coexpression of all the subunits completely eliminated the stimulatory effect of (-)D600, while at the same time increasing the affinity of the complex for DHP to that stabilized in partial complexes by the phenylalkylamine. These results demonstrate that all of the components that co-purify are required for the formation of a functional DHP receptor having the properties of the native skeletal muscle DHP receptor.

Animals↗
PubMed1996

Increase of the expression of midecamycin 4"-hydroxyl propionyltransferase gene (mpt) by a promoter-like fragment from the midecamycin producing strain.

The promoter region of the midecamycin 4"-hydroxyl propionyltransferase gene (mpt) gene was reconstructed by PCR, and ligated with a fragment from the midecamycin producing strain (Streptomyces mycarofaciens var. 68) which contained fairly strong promoter activity (PLF). Recombinant plasmids pCHFPE2 (promoter region of mpt was reconstructed by combining the PLF in the upstream of its own promoter) and pCHFPE3 (the promoter of mpt was replaced by PLF) were obtained. The extent of expression of mpt was measured according to the amount of propionylspiramycin bioconverted from exogenous spiramycin by transformants of S. lividans TK24 containing pCHFPE2 and pCHFPE3. The results showed that the PLF could increase the expression of mpt in S. lividans TK24 up to 89.02% and 58.53%, respectively, and also enhance the expression in the industrial spiramycin producing strain S. spiramyceticus to a great extent.

Acyltransferases↗
PubMed1996

[Microsurgical anatomical study of the wall of the cavernous sinus].

OBJECTIVE: To observe the microanatomical structures of the wall of cavernous sinus (CS) in order to provide anatomical basis for direct microsurgical operation to the CS. METHODS: 48 aides of the CS of adult cadavers were dissected in detail under operative microscope. RESULTS: The CS was a narrow and long irregular hexahedron in shape. The supperior wall, the posterior wall and the superficial dural layer of the lateral wall of the CS were formed by internal layer of the cerebral dura. The deep layer of the interal wall was formed by the sheaths of nerves II, IV, V1, V2 with a reticular membrane between these sheaths. There was a superficial compartment between the superficial layer and deep layer of the lateral wall in 31.25% of the specimens. The inferior wall of the CS was intracranial periosteum. The medial wall was formed by stretched fibres from the tentorium of hypophysis. The anterior aspect was formed by the sphenoid sinus and the superior orbital fissure. Direct microsurgical approaches to the CS via the medial trangles of the superior wall and Parkinson tranales of the lateral wall were most important. CONCLUSION: A microanatomical structure of the wall of the CS is provided for direct microsurgical operation to the CS, and the direct microsurgical approaches are discussed.

Adult↗
PubMed1996

Prevention of disability and rehabilitation--results from a collaborative project in China.

Eight different geographical districts in China varying in urban and rural characteristics from 6 provinces and 2 municipalities were selected as pilot areas of the project. The interventions included early detection and treatment of neuritis, self-care training, adapted footwear, surgery, comprehensive treatment of complicated ulcers, and prostheses. Main changes in eye, hand and foot impairments between baseline assessment and assessment at 2 years have been presented in this paper.

China↗
PubMed1996

[Antagonistic effect of tetrahydroproberberine homologues on alpha 1-adrenoceptor].

The antagonistic effect of tetrahydroproberberine (THP) homologues on alpha 1-adrenoceptor was studied by combination of radioligand binding assays and measurements of vasoconstriction responses. The results showed that l-tetrahydropalmatine (l-THP), l-stepholidine (l-SPD), THPB-18 and tetrahydroberberine (THB) competitively inhibited the 125I-BE2254 specific binding in rat cerebral cortex with pK1 values of 5.54 +/- 0.36, 5.56 +/- 0.47, 5.75 +/- 0.56 and 6.01 +/- 0.60, respectively, and the Hill efficiency was not significantly different from unity. They inhibited phenylephrine-induced constrictions with pA2 values of 5.48 +/- 0.58, 5.66 +/- 0.54, 5.64 +/- 0.34 and 5.45 +/- 0.76, respectively, and the slopes of Schild plot were not significantly different from unity. The results indicate that the 4 THP homologues are non-subtype selective competitive antagonists for alpha 1-adrenoceptor with similar affinities.

Adrenergic alpha-Antagonists↗
PubMed1995

Essential Ca(2+)-binding motif for Ca(2+)-sensitive inactivation of L-type Ca2+ channels.

Intracellular calcium (Ca2+) inhibits the opening of L-type (alpha 1C) Ca2+ channels, providing physiological control of Ca2+ entry into a wide variety of cells. A structural determinant of this Ca(2+)-sensitive inactivation was revealed by chimeric Ca2+ channels derived from parental alpha 1C and alpha 1E channels, the latter of which is a neuronal channel lacking Ca2+ inactivation. A consensus Ca(2+)-binding motif (an EF hand), located on the alpha 1C subunit, was required for Ca2+ inactivation. Donation of the alpha 1C EF-hand region to the alpha 1E channel conferred the Ca(2+)-inactivating phenotype. These results strongly suggest that Ca2+ binding to the alpha 1C subunit initiates Ca2+ inactivation.

Amino Acid Sequence↗
PubMed1995

Molecular determinants of cardiac Ca2+ channel pharmacology. Subunit requirement for the high affinity and allosteric regulation of dihydropyridine binding.

Cardiac L-type Ca2+ channels are multisubunit complexes composed of alpha 1C, alpha 2 delta, and beta 2 subunits. We tested the roles of these subunits in forming a functional complex by characterizing the effects of subunit composition on dihydropyridine binding, its allosteric regulation, and the ability of dihydropyridines to inhibit channel activity. Transfection of COS.M6 cells with cardiac alpha 1C-a (alpha 1) led to the appearance of dihydropyridine ([3H]PN200-110) binding which was increased by coexpression of cardiac beta 2a (beta), alpha 2 delta a (alpha 2), and the skeletal muscle gamma. Maximum binding was achieved when cells expressed alpha 1, beta, and alpha 2. Cells transfected with alpha 1 and beta had a binding affinity that was 5-10-fold lower than that observed in cardiac membranes. Coexpression of alpha 2 normalized this affinity. (-)-D600 and diltiazem both partially inhibited PN200-100 binding to cardiac microsomes, but stimulated binding in cells transfected with alpha 1 and beta. Again, coexpression of alpha 2 normalized this allosteric regulation. Therefore coexpression of alpha 1 beta and alpha 2 completely reconstituted high affinity dihydropyridine binding and its allosteric regulation as observed in cardiac membranes. Skeletal muscle gamma was not required for this reconstitution. Expression in Xenopus oocytes demonstrated that coexpression of alpha 2 with alpha 1 beta increased the potency and maximum extent of block of Ca2+ channel currents by nisoldipine, a dihydropyridine Ca2+ channel antagonist. Our results demonstrate that alpha 2 subunits are essential components of the cardiac L-type Ca2+ channel and predict a minimum subunit composition of alpha 1C beta 2 alpha 2 delta for this channel.

Allosteric Regulation↗