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Biomedical subjects

X M Wang

Publications and source records attributed to X M Wang.

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

Disruption of adhC reveals a large duplication in the Mycobacterium smegmatis mc(2)155 genome.

Disruption of the adhC gene of Mycobacterium smegmatis mc(2)155, by standard gene replacement methods, revealed that there are two copies of this gene within a large duplication of the M. smegmatis mc(2)155 genome. M. smegmatis AdhC(+/-) and M. smegmatis AdhC(-/-) mutants were obtained when one or two adhC copies, respectively, were disrupted by homologous recombination. Southern blot analysis of DraI restriction digests of the DNA from these mutants and from wild-type M. smegmatis mc(2)155, resolved by PFGE, showed that the duplication size may be at least approximately 250 kb. The single and double knockout mutants were characterized and compared with the M. smegmatis wild-type. A growth disadvantage and a different morphology were associated with the loss of expression of one or both of the adhC copies, but both mutants were still acid-fast. Findings in this study indicate that the process of chromosomal duplication in M. smegmatis is ongoing and remains a potent source of genome dynamics. Hence, the M. smegmatis mc(2)155 genome might be larger than previously thought.

Alcohol Dehydrogenase↗
PubMed2001

Scattering of Lamb waves by a circular cylinder.

Following our previous attempt at the scattering from a cylinder in a slab to the incidence of a guided shear wave, we hereby discuss the scattering by an elastic cylinder embedded in an isotropic plate for a variety of bonding states to incidence of the fundamental Lamb wave modes S0 and A0 at the low-frequency regime. The plate is divided up into three regions by introducing two imaginary planes located symmetrically some distance from the cylinder and perpendicular to surfaces of the plate. The wave fields in various regions are expanded either into cylinder wave modes or into Lamb wave modes. A system of equations determining the coefficients of expansion is obtained according to the traction-free boundary conditions on the plate walls and the displacement and stress continuity conditions across the virtual planes. By taking an appropriate finite number of terms of the infinite expansion series and some selected points on the two properly chosen imaginary planes and the surfaces of the plate through convergence and precision tests, a matrix equation to numerically evaluate the expansion coefficients is found. Coefficients of the reflection and transmission versus the normalized radius of the cylinder in welded, slip, and cracked interfacial conditions are numerically computed. In the low-frequency range, the relative errors are found to be less than 1%. Contrast curves of the reflection coefficient for the cylinder of nearly all permissible size in perfect and imperfect interfacial bonding are shown and prominent differences are noted.

Journal Article↗
PubMed2001

[Generation of Mx-cre transgenic mice].

To generate the transgenic mice expressing cyclization recombination enzyme, the recombinant gene, in which the coding region of cre gene is derived by the promoter of mouse Mx gene, was microinjected into pronuclei of fertilized mouse eggs. Founders of transgenic mice harbouring the recombinant gene were screened by polymerase chain reaction (PCR) at genomic DNA level and confirmed by Southern blot. One line of Mx-cre transgenic mice was obtained. Then, the Mx-cre transgenic mouse line was cultured and propagated.

Animals↗
PubMed2001

[Expression of Trial in mammalian cell and its apoptosis activation in culture cells].

To develop a system for expression of Trail in mammalian cell. PCR and sequencing were used to clone and to confirm Trail gene. Then, we constructed the vector for expression of Trail in mammalian cells. HeLa cells, one of sensitive cell lines to Trail, were transfected with the vector DNA. After 48 h, flow cytometry was used to screen HeLa transfected by Trail. 19% of total cells happened to apoptosis. This study conforms that the production of Trail expressing in mammal cell can induce apoptosis and exhibit high bio-active. The results indicate the potential application of Trail to tumor immunotherapy.

Apoptosis↗
PubMed2001

[The presentation of neo gene in chimeric mice obtained from ES cells].

To explore the potentiality of the mouse ES cell line which is employed in our laboratory were transfected with neo gene and selected with G418. Some ES cell clones expressing neo gene were obtained. After ES clones were picked up and expanded, the ES cells were mircoinjected into blastocysts and implanted miceuterus. 60 blastocysts were implanted into 5 mice. Finally, 3 chimeric mice with neo gene were born. PCR was carried out to analyze the presentation of neo gene in tissues of chimeric mice. The results showed that many tissues harbored the neo gene, including skin, liver and blood et al.

Animals↗
PubMed2001

[Human dynamic response to landing impact in selected body orientations].

Objective. To study changes of human dynamic response to landing impact in selected body orientations. Method. Five healthy young men were exposed to 10 G, 50 ms half-sine acceleration pulses. The angle included between human head-ilium axis and impact force direction was 30 degrees-70 degrees. The acceleration responses at head, shoulder, chest and ilium of the subjects were recorded along head-ilium and chest-back directions. Result. Within limits of the experiments, the minimum peak response at the head along head-ilium direction was 10.36 +/- 2.44 G at 50 degrees, and the maximum was 18.07 +/- 3.29 G at 70 degrees. The minimum peak response at the chest along head-ilium direction was 10.39 +/- 3.97 G at 70 degrees, and the maximum was 15.42 +/- 3.61 G at 60 degrees. The minimum peak response at the head along chest-back direction was 7.58 +/- 1.18 G at 30 degrees, and the maximum was 18.89 +/- 1.85 G at 70 degrees. The minimum peak response at the chest along chest-back direction was 7.21 +/- 1.99 G at 30 degrees, and the maximum was 17.67 +/- 2.16 G at 70 degrees. Conclusion. It was inferred that the best body orientation angle for landing impact might be 50 degrees.

Acceleration↗
PubMed2001

[Separation of xylose and xylitol in fermentation liquid by capillary zone electrophoresis].

Xylitol may be produced with microbial fermentation technology when xylose is used as the raw material. It is important in scientific research and production fields concerned with xylose and xylitol to develop the method of separation and detection. The anionic complexes with strong UV absorption at 195 nm will form by dissolving xylose and xylitol in borax solution. They may be separated with borax buffer by use of capillary zone electrophoresis. It was shown that the resolution between xylose and xylitol gradually increased with the increase of borax concentration, but its maximum concentration was 130 mmol/L at room temperature. The resolution depended on the pH of running buffer with a maximum at pH 9.55. The resolution was independent of cetyltrimethylammonium bromide when its concentration was between 4 x 10(-6) mmol/L-8 x 10(-4) mmol/L. So the optimum conditions were as follows: borax concentration of 130 mmol/L, pH 9.55 and cetyltrimethylammonium bromide concentration of 5 x 10(-5) mmol/L in running buffer; separating voltage of -12 kV; column temperature of 25 degrees C. Xylose and xylitol could be separated on base line in 6 min under these conditions. Samples from fermentation process and the recoveries of spiked samples were determined. The relative standard deviations of the results were between 1.42%-3.11% for xylose, and 0.62%-1.32% for xylitol. The recoveries were between 96.0%-108.0% for xylose and 94.0%-109.0% for xylitol.

Electrophoresis, Capillary↗
PubMed2001

[1.4 kb 5' flanking region of class I patatin directs tuber-specific gus expression in potato (Solanum tuberosum L.)].

Binary vectors pPATIs (with partial signal sequence) and pPATI (without signal sequence) were constructed by fusing 1.4 kb 5' flanking regions of Class I patatin gene with GUS. Transient GUS expression was observed in in vitro tuber slices bombarded with pPATI. These constructs were then introduced into potato (cv. Desiree) via Agrobacterium tumefaciens transformation. Transgenic potato plants were confirmed by X-Gluc staining and PCR. Using in vitro tuberization system, GUS activities were assayed by fluorescence. It was shown that, in plants transformed with PATI-GUS, GUS specific activities were about 10-20 fold higher in tubers than in stems. Increased sucrose concentration could not induce PATI-GUS expression, but light enhanced PATI-GUS expression in cultured shoots.

Carboxylic Ester Hydrolases↗
PubMed2001

[Effect of wuzi yanzong pill and its disassembled prescription on mitochondrial DNA deletion, respiratory chain complexes and ATP synthesis in aged rats].

OBJECTIVE: To investigate the effect of Wuzi Yanzong Pill (WZYZP) and its disassembled prescription on mitochondrial DNA deletions, respiratory chain complexes and ATP synthesis in aged rats. METHODS: Animal experiments with polymerase chain reaction (PCR), enzyme kinetics and bioluminescence technique were conducted. RESULTS: WZYZP and its disassembled prescription of Fructus Lycii and Semen Cuscutae could reduce the mitochondrial DNA deletions and raise the activities of mitochondrial respiratory chain complexes I, IV and adenosine triphosphate (ATP) synthesis in aged rats' brain; Fructus Lycii and Semen Cuscutae could also reduce the mitochondrial DNA deletions in aged rats' heart (P < 0.05, P < 0.01). CONCLUSION: WZYZP, Fructus Lycii and Semen Cuscutae have protective effect on oxidative damage of mitochondrial DNA in aged rats.

ATP Synthetase Complexes↗
PubMed2001

[Selective isolation of anethole from volatile oil of Foeniculum vulgare Mill by inclusion crystalline with chela-shape host].

AIM: To isolate the components from the volatile oil of Foeniculum vulgare Mill. METHODS: According to the function of molecular recognition of supramolecular chemistry, chela shape molecule, trans-1, 2-biphenyl-1, 2-acenaphthendiol was used as host molecule and the volatile oil of Foeniculum vulgare Mill as guest molecule. Trans-1, 2-biphenyl-1, 2-acenaphthendiol can recognize the components that endowed with interactional complementarity and form inclusion compound as crystals. RESULTS: The anethole in the volatile oil was selectively included as trans-1,2-biphenyl-1,2-acenaphthendiol which was obtained in pure state from the inclusion compound by Kugelrohr vacuum technology. The formation of inclusion compound was confirmed by means of IR and powder XRD. The structure of the selectively isolated component was elucidated as trans-anethole by means of IR, 1HMMR and MS. CONCLUSION: The experimental results showed that the method is simple, rapid and selective for isolation anethole from volatile oil of Foeniculum vulgare Mill.

Allylbenzene Derivatives↗
PubMed2000

Regeneration of descending spinal axons after transection of the thoracic spinal cord during early development in the North American opossum, Didelphis virginiana.

Opossums are born in an immature, fetal-like state, making it possible to lesion their spinal cord early in development without intrauterine surgery. When the thoracic spinal cord of the North American opossum, Didelphis virginiana, is transected on postnatal day 5, and injections of Fast Blue (FB) are made caudal to the lesion site 30-40 days or 6 months later, neurons are labeled in all of the spinal and supraspinal areas that are labeled after comparable injections in age-matched, unlesioned controls. Double-labeling studies document that regeneration of cut axons contributes to growth of axons through the lesion site and behavioral studies show that animals lesioned on postnatal day 5 use their hindlimbs in normal appearing locomotion as adults. The critical period for developmental plasticity of descending spinal axons extends to postnatal day 26, although axons which grow through the lesion site become fewer in number and more restricted as to origin with increasing age. Animals lesioned between postnatal day 12 and 26 use the hindlimbs better than animals lesioned as adults, but hindlimb function is markedly abnormal and uncoordinated with that of the forelimbs. We conclude that restoration of anatomical continuity occurs after transection of the spinal cord in developing opossums, that descending axons grow through the lesion site, that regeneration of cut axons contributes to such growth, and that animals lesioned early enough in development have relatively normal motor function as adults.

Animals↗
PubMed2000

[Effects of different loading exercises on endogenous sex hormones and their metabolites].

Small loading and strengthening exercises were performed by 22 athletes(11 males and 11 females). The levels of serum hormones and their urinary metabolites were detected. The results showed that the serum levels of testosterone(T), free testosterone(FT) and 17-OH-progesterone(17-OHP) in male athletes exhibited no conspicuous changes. The excretion of their urinary androsteroids was significantly increased after exercises(P < 0.05), but significantly decreased in the recovery period(P < 0.05). After small loading exercises, their serum levels of T and FT revealed no obvious changes except those of 17-OHP which evidently increased in female athletes(P < 0.05); whereas the excretion of their urinary steroid hormones declined promptly after the exercises and in the recovery period (all P < 0.05). After strengthening exercises, the serum levels of T, FT and 17-OHP decreased distinctly in female athletes (all P < 0.05), the excretion of their urinary steroid hormones raised clearly after training and in the recovery period(P < 0.05). Therefore, it is suggested that the secretion and metabolism of sex hormones in the organism are influenced by the loading quality of sports during training, and sexual difference exists simultaneously.

17-alpha-Hydroxyprogesterone↗
PubMed2000

HPV11 mutant virus-like particles elicit immune responses that neutralize virus and delineate a novel neutralizing domain.

Characterization of the regions of human papillomaviruses (HPVs) that elicit neutralizing immune responses supports studies on viral infectivity and provides insight for the development and evaluation of prophylactic vaccines. HPV11 is a major etiologic agent of genital warts and a likely vaccine candidate. A conformationally dependent epitope for the binding of three neutralizing monoclonal antibodies (mAbs) has been mapped to residues G(131)T(132) of the L1 major capsid protein. The mAbs bind L1 only when it is assembled into virions or into virus-like particles (VLPs) that mimic the capsid structure. We were interested in identifying other domains of L1 that elicit neutralizing responses. To this end, we have generated a panel of mAbs against VLPs derived from HPV11 L1 harboring a G131S substitution. The new mAbs are unlike the neutralizing mAbs previously mapped to residues G(131)T(132) in that they bind both prototype and HPV11:G131S mutant VLPs. Some of the new mAbs neutralized virus in vitro. We have mapped epitopes for three of these new mAbs, as well as a neutralizing mAb generated against HPV11 virions, by measuring binding to HPV6 VLPs substituted with HPV11-like amino acids. Two regions are critical: one defined by HPV11 L1 residues 263-290 and the other by residues 346-349. mAbs H11.H3 and H11.G131S.G3 bind HPV6 VLPs with substitutions derived from the 346-349 region; in addition, H11.G131S.G3 binds HPV6 VLPs with substitutions derived only from the 263-290 region. Although H11.H3 does not bind HPV6 VLPs with substitutions derived from the 263-290 region, binding to HPV6 VLPs is enhanced when both sets of substitutions are present. mAbs H11.G131S.I1 and H11.G131S.K5 bind HPV6 VLPs with the 263-290 substitutions, but show little binding to HPV6 VLPs with the 346-349 substitutions. However, binding to HPV6 VLPs is enhanced when substitutions at both regions are present. The 346-349 region has not previously been described as eliciting a neutralizing response for any HPV type. In addition, the work demonstrates a complex binding site contributed by two distinct regions of L1.

Animals↗
PubMed2000

Inhibition of platelet aggregation by the recombinant cysteine-rich domain of the hemorrhagic snake venom metalloproteinase, atrolysin A.

The P-III class of venom metalloproteinases has, in addition to the proteinase domain, a disintegrin-like domain and a cysteine-rich domain. Recent evidence has shown that the nonproteinase domains of the P-III class of hemorrhagic metalloproteinases function in the inhibition of platelet aggregation by blocking essential procoagulant integrins on platelets. A specific role for the highly conserved cysteine-rich domain has yet to be described. In this study, we expressed the cysteine-rich domain from the hemorrhagic metalloproteinase atrolysin A and demonstrated its ability to inhibit collagen-stimulated platelet aggregation. Additionally, the cysteine-rich domain was shown to interact with MG-63 cells to inhibit adhesion to collagen I. These data suggest a functional role for the cysteine-rich domain of the P-III toxins in the observed coagulopathy by targeting the toxin to platelets and inhibiting collagen-stimulated platelet aggregation. These characteristics may function to synergistically increase the hemorrhagic effect of the toxins.

Amino Acid Sequence↗
PubMed2000

Repair of the transected spinal cord at different stages of development in the North American opossum, Didelphis virginiana.

When the thoracic spinal cord is transected on postnatal day (PD) 5 in the North American opossum, descending and ascending axons grow through the lesion site. When the lesion is made on PD20, comparable growth is limited to a subset of descending axons. To better understand the mechanisms underlying these differences, we analyzed the transection site at different times after lesioning at both ages. Axons which crossed the lesion site could be identified using silver impregnation and immunostaining for neurofilament. Nissl stains revealed that abnormal appearing grey matter was also present in some of the PD5 cases. In many PD5 cases, however, and in all of the animals transected at PD20, grey matter was not present at the lesion site. Immunostaining with a neuron specific antibody supported that conclusion. However, immunostaining with phenotypic specific antibodies revealed that glial cells were present in all cases. Immunostaining for Schwann cells was negative. Fibronectin-positive cells were also present at the lesion site after transection of the thoracic cord at PD20, but their identity was uncertain. When injections of bromodeoxyuridine (BrdU), a thymidine analog, were made at different times after lesioning and the pups were sacrificed for BrdU immunohistochemistry up to 40 days later, labeled cells were found in the tissue which bridged the lesion site indicating that cell proliferation contributed to reconstruction at the lesion site.

Age Factors↗
PubMed2000

Structure and interaction of VacA of Helicobacter pylori with a lipid membrane.

In its mature form, the VacA toxin of Helicobacter pylori is a 95-kDa protein which is released from the bacteria as a low-activity complex. This complex can be activated by low-pH treatment that parallels the activity of the toxin on target cells. VacA has been previously shown to insert itself into lipid membranes and to induce anion-selective channels in planar lipid bilayers. Binding of VacA to lipid vesicles and its ability to induce calcein release from these vesicles were systematically compared as a function of pH. These two phenomena show a different pH-dependence, suggesting that the association with the lipid membrane may be a two-step mechanism. The secondary and tertiary structure of VacA as a function of pH and the presence of lipid vesicles were investigated by Fourier-transform infrared spectroscopy. The secondary structure of VacA is identical whatever the pH and the presence of a lipid membrane, but the tertiary structure in the presence of a lipid membrane is dependent on pH, as evidenced by H/D exchange.

Bacterial Proteins↗
PubMed2000

Geographic clustering of an outer surface protein A mutant of Borrelia burgdorferi. Possible implications of multiple variants for Lyme disease persistence.

DNA sequences encoding full-length outer surface protein (Osp) A were amplified from four joint fluid samples over 4.5 months from a patient with chronic Lyme arthritis, with a variant from wild type only found in sample 3. Rather than a mutation in vivo, these findings suggested a mixed infection in which BORRELIA: containing the wild-type and mutant ospA were waxing and waning in the patient's joint. If so, we reasoned that the mutant should be present in the community. We therefore took the novel epitope resulting from the mutation, expressed as a fusion protein in Escherichia coli, and performed Western blots on 80 high-titred stored sera; however, all except that of our index patient were negative. We then collected 36 stored sera from patients with Lyme disease residing within 10 miles of where the index patient had lived. An additional two sera from this circumscribed area were positive (P = 0.038). These findings show that results from single samples can be misleading, and suggest that the OspAs expressed in force late in Lyme arthritis are the same ones introduced initially into the host. Moreover, they allow a speculative mechanism for disease persistence not previously considered, in which antigenically distinct B. burgdorferi variant proteins present themselves serially to the immune system.

Antigens, Surface↗