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Biomedical subjects

X Chu

Publications and source records attributed to X Chu.

At least 55 records · Page 3Linked to original sources

[Purification and some properties of superoxide dismutase from Fusarium moniliform].

Superoxide dismutase from Fusarium moniliform was purified by the steps including heating, ammonium sulfate fractionation, Sephedax G-100 gel filtration and DEAE-Sephadex A-50 chromatography. The results showed that the enzyme was a Mn-SOD with the specific activity of 2640 U/mg and had two homogenous subunits whose molecular mass were 14.5 kD. The wave length of max. absorbing peak in ultraviolet spectrum was 276 nm which was not similar with other resource of SOD. The composition of amino acid was also analyzed.

Amino Acids↗

[Studies on the changes of morphometry and neuropeptide of spinal neurons after peripheral nerve injury].

To observe the change of morphology and neuropeptide in the spinal neurons in order to clarify the functional state after injury of peripheral nerves is especially in the late stage. Sciatic nerves were cut with their proximal segments in the preparation of a model of peripheral nerve injury. Combination of horseradish peroxidase retrograde tracing immunohistochemistry and computer image analysis the changes in the morphometry of the perikarya of ventral horn neurons of the spinal cord, the quantitative changes of substance P (SP). Calcitonin gene-related peptide (CGRP) in dorsal horn and CGRP and choline acetyransferase (CHAT) in ventral horn of the spinal cord were examed. The results showd: (1) At the 3rd week after injury, swollen perikarya of the ventral horn neurons were observed, subseauently the swelling of perikarya was decreased tile the 6th week the neurons recovered to their normal size. At the 12th week the neurons were generally stable in their size, shortening of the dendrites was seen in 27% of the neurons. (2) The dendrites of the neurons progressively contracted till at the 12th week 53% of them were degenerated. The results of the 24th week were similar to the that at the 12th week. (3) CGRP in the ventral horn of the spinal cord was elevated to the highest point after 1 week of injury, that lasting for 4 weeks and 8 weeks later, the lever of CGRP returned to normal. From 20th to 24th week, there was no obvious changes of CHAT in the ventral horn of the spinal cord during observation. (4) SP went to the lowest point in the dorsal horn during 2-6 weeks, then recovered slowly, and beiny normal again after 16 weeks, however, CGRP was changed slightly. The results indicated that although a series of degenerating changes occurred in the neurons of the spinal cord during the late peripheral nerve injury, but the functional activity of the central meurons still was maintained at a certain level.

Animals↗

[Peripheral nerve injury as a complication from orthopedic operation].

Nerve injury following operation is one of the main causes of the iatrogenic peripheral nerve injury. In order to learn lessons from these cases, one hundred and seven cases of peripheral nerve injury complicated with the orthopedic operations were analyzed. Forty-four cases were cutting injury during operation, made up 41% of all cases and 27 cases were stretch and compression injury, made up 25%. The involved nerves included 41 radial nerves and 24 common peroneal nerves, composing 60.7% of all nerve injury. The operations responsible were mainly the bone and joint operations, which made up 81%. The cause, prophylaxis, diagnosis and treatment were discussed. The rich appropriate knowledge of anatomy and responsibility of the surgeon were emphasized in order to prevent the occurrence of complication. Once the injury was suspected, diagnosis should be made promptly and effective treatment should be performed in time.

Adolescent↗

Attractive Interaction between Similarly Charged Colloidal Particles

The pair interactions between the charged colloidal particles dispersed in a solvent are studied theoretically by the integral equation method. The pair potential of the mean forces, accounting for the effective pair interaction between colloidal particles, is calculated from the solution of the Ornstein-Zernike equation with the mean spherical approximation (MSA). An attractive interaction was found between two similarly charged colloidal particles in contrast with the purely repulsive force predicted by the Debye-Huckel theory. Such an attractive interaction provides physical insight for the "condensed" phenomena in charged colloidal dispersions, that is, the coexistence of a "condensed" phase and an "expanded" phase (voids). At the higher concentration and charge on colloidal particles, the effective pair interaction becomes oscillatory.

Journal Article↗

Polymer agglutination-based piezoelectric immunoassay for the determination of complement III.

A piezoelectric immunoassay technique, which is based on the detection of agglutination of antibody- or antigen-bearing polymer by an immunoreaction using a piezoelectric quartz crystal, has been developed for the determination of complement III (C3). Anti-C3 antibodies were physically adsorbed onto the carboxymethyl cellulose polymer by hydrogen bonds. The agglutination of the (anti-C3 antibody)-bearing polymer by immunoreaction caused a viscosity change of the solution, which could be monitored by a piezoelectric quartz crystal. The effect of experimental conditions such as the polymer concentration, the antibody dilution ratio and the reaction temperature on the frequency response were investigated. The linear ranges for C3 concentration determined by the end-point method and the initial rate method were 22.0-43.2 micrograms ml-1 and 22.0-49.1 micrograms ml-1, respectively. Other antigens presenting in serum did not interfere with the determination of C3. Analytical results of ten clinical specimens obtained using the developed technique were in satisfactory agreement with those given by the rate diffusion turbidimetry. With a simple regeneration method devised, the crystal can be used repeatedly with acceptable reproducibility.

Complement C3↗

Identification of Streptomyces violaceoruber Tü22 genes involved in the biosynthesis of granaticin.

A 50 kb region of DNA from Streptomyces violaceoruber Tü22, containing genes encoding proteins involved in the biosynthesis of granaticin, was isolated. The DNA sequence of a 7.3 kb fragment from this region, located approximately 10 kb from the genes that encode the polyketide synthetase responsible for formation of the benzoisochromane quinone skeleton, revealed five open reading frames (ORF1-ORF5). The deduced amino acid sequence of GraE, encoded by ORF2, shows 60.8% identity (75.2% similarity) to a dTDP-glucose dehydratase (StrE) from Streptomyces griseus. Cultures of Escherichia coli containing plasmids with ORF2, on a 2.1 kb BamHI fragment, were able to catalyze the formation of dTDP-4-keto-6-deoxy-D-glucose from dTDP-glucose at 5 times the rate of control cultures, confirming that ORF2 encodes a dTDP-glucose dehydratase. The amino acid sequence encoded by ORF3 (GraD) is 51.4% identical (69.9% similar) to that of StrD, a dTDP-glucose synthase from Streptomyces griseus. The amino acid sequence encoded by ORF4 shares similarities with proteins that confer resistance to tetracycline and methylenomycin, and is suggested to be involved in transporting granaticin out of the cells by an active efflux mechanism.

Amino Acid Sequence↗

Polymerase chain reaction analysis of the Xba I polymorphism of the human complement C4 genes provides evidence for strong haplotype conservation.

The genes coding for the two isotypes of the fourth component of human complement, C4A and C4B, are located between the HLA-B and -DR loci of the MHC. We studied the linkage relationship of the previously described XbaI RFLP to obtain further insight into the evolution of the tandemly arranged C4 genes. Using exon-specific PCR amplification followed by restriction analysis and direct DNA sequencing, the polymorphic site could be located in exon 40 of the C4 gene (cDNA position 5095). The polymorphism does not change an amino acid residue. Using nested PCR amplification with isotype-specific primers to amplify either C4A or C4B alleles the haplotype arrangement of the XbaI sites in both isotypic C4 genes was analyzed independently. It was observed that the XbaI restriction site was either present or absent in both C4 genes of a given haplotype. In a study of 106 Caucasian haplotypes, only two different haplotypes could be identified carrying a C4A gene with and a C4B gene without the XbaI restriction site. Also, the XbaI site could only be detected in long C4 genes possessing the 6.5-kb insertion in intron 9. Our findings provide evidence that the mutation creating the XbaI polymorphism occurred in an ancestral C4 gene already carrying the long intron 9. The duplicating resulting in the presence of two isotypic genes, C4A and C4B, must have taken place subsequently giving rise to haplotypes with or without the XbaI site.

Base Sequence↗

Piezoelectric immunosensor for the detection of immunoglobulin M.

A piezoelectric immunosensor has been developed for the determination of human IgM. The crystals are AT-cut and have a basic resonant frequency of 9 MHz. Immobilization of goat antihuman IgM antibodies to the crystals' surfaces was accomplished via a CNBr-activated copolymer coating of 2-hydroxyethyl methacrylate and methylmethacrylate. The IgM piezoelectric immunosensor can be used for the human IgM determination in the range 5-93 micrograms ml-1. The analytical results given by this approach were in satisfactory agreement with those given by the single radical immunodiffusion procedure. The sensitivity, specificity and reproducibility of this immunosensor were investigated. Further, the valent value of goat antihuman IgM antibody binding with human IgM antigen and the affinity constant of immunoreaction in this experimental system were studied. After washing with tetrahydrofuran, a crystal can be re-used 20 times without detectable loss of sensitivity.

Biosensing Techniques↗

St14 (DXS52) VNTR in the Chinese population and its application to genetic diagnosis of haemophilia A.

The variable number of tandem repeats (VNTR) of St14 (DXS52) on the human X-chromosome was analysed using the polymerase chain reaction (PCR) method. Screening of 78 X-chromosomes in 56 healthy Chinese individuals revealed the existence of at least seven different alleles in the the Chinese population, the corresponding amplified fragments and frequencies being 700 bp (60.3%), 1220 bp (1.3%), 1300 bp (2.6%), 1390 bp (11.5%), 1570 bp (12.8%), 1630 bp (6.4%) and 1690 bp (5.1%). Total theoretical heterozygous rate was 60%. Compared to Caucasians, this Chinese population showed a markedly higher occurrence of low molecular weight fragments and a relatively low occurrence of high molecular weight fragments. Study of this polymorphism in 14 suspected haemophilia A carriers revealed half of them to be heterozygous. Thus, St14 VNTR analysis by PCR should prove to be a useful tool in the genetic diagnosis of haemophilia A in China.

Asian People↗

Length polymorphism of the human complement component C4 gene is due to an ancient retroviral integration.

The fourth component of the complement system, C4, is encoded by two highly homologous MHC-linked genes expressing the two isotypes C4A and C4B. A gene size polymorphism (either 22.5 or 16 kb) has been described which depends on the presence or absence of a 6.5-kb insertion in intron 9 of the C4 gene. By sequencing a C4A-specific lambda clone from a human genomic library containing the long intron 9 as well as PCR-amplified DNA containing the short intron, the DNA sequences of both introns were determined. The long and short introns have lengths of 6,787 bp and 415 bp, respectively. The sequence of the short intron is almost identical (96%) to the corresponding parts of the long intron. At position 282 of the short intron, a 6,372-bp insertion is present in the long intron which has all characteristics of a full-length endogenous retrovirus. The proviral DNA is flanked by two 6-bp target site repeats. The orientation of the proviral sequence is opposite to that of the C4 coding strand. Long terminal repeats (LTRs) of 548 bp were found at both ends of the provirus. A TATA box and an SV40 enhancer core as well as a polyadenylation signal are present in the LTR. A 5' primer binding site for lysine tRNA was identified. The strongest sequence homologies were found in comparison to human endogenous retrovirus (HERV-K): between 65-88% for gag, pol and env genes. However, a search for open reading frames in these regions indicated the presence of multiple stop codons in all three reading frames.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Composition↗

[Site-directed mutagenesis of Lac Z gene in Escherichia coli and the kinetic properties of the mutated enzymes].

Glutamic acid at position of 537 of beta-D-galactosidase coded by Lac Z gene was substituted with Aspartic acid, Glutamine and Valine using synthetic oligonucleotide probes. Compared to native enzyme, the kcat values for substrate ONPG were 0.13%, 0.0006% and 0.0035% for Asp-537, Gln-537 and Val-537 mutated enzymes respectively. The Km values were of the same order of magnitude, either native or mutated enzymes. The substrate analog, IPTG was a strong inhibitor of each of the substituted enzymes, as in the case of native enzyme. The transition state analogs, 2-NH2-galactose and L-ribose were almost the same effects for the mutated enzymes as for the normal enzyme. The nucleophili, Azide, did not activate the mutated enzymes as in the case of Glu-461 substituted in beta-D-galactosidase. The effect of methanol on the mutated enzymes was less than on native enzyme. The order of the thermal stability was native enzyme > Asp-537 > Gln-537 > Val-537 enzymes. Overall, the evidence strongly supports the suggestion that Glu-537 is an essential residue of beta-D-galactosidase.

Base Sequence↗

Defibrotide stimulates expression of thrombomodulin in human endothelial cells.

Cultured human umbilical vein endothelial cells were incubated with defibrotide at concentrations of 0, 5, 50 and 500 micrograms/ml for 4 and 24 h respectively. Thrombomodulin activity and molecules on the surface of the cells were determined by chromogenic assay and radioimmunoassay, thrombomodulin antigen in endothelial cells and in conditioned medium of the cells was measured by immunoradioassay. Thrombomodulin mRNA within the cells was analysed by slot blot. After 24 h of incubation, the activity and molecules of thrombomodulin on the surface of endothelial cells, as well as the antigen and mRNA of thrombomodulin in the cells were significantly increased in a dose dependent manner. However, the level of thrombomodulin antigen in conditioned medium was about equal to that of the control. Our data indicate that defibrotide stimulates expression of thrombomodulin in human endothelial cells. These beneficial effects may play a role in antithrombotic activity of defibrotide.

Cells, Cultured↗

Application of GPIIIa gene Taq I polymorphism to determination of carrier status in Glanzmann's thrombasthenia families of Chinese origin.

Glanzmann's thrombasthenia is a bleeding disorder caused by qualitative and/or quantitative defects of platelet membrane glycoprotein (GP) IIb/IIIa complex. The disease is inherited in an autosomal recessive manner. In this paper, cDNA probes were used to study restriction fragment length polymorphisms (RFLPs) in GPIIIa gene. A Taq I polymorphism was identified and this RFLP was composed of variant bands of 6.5 Kb/4.0 and 2.5 Kb with a frequency of 0.46/0.54 in Chinese population. The Taq I polymorphism was further localized by polymerase chain reaction (PCR) method to exon VIII of the GPIIIa gene. In two Glanzmann's thrombasthenia families, the Taq I RFLP studied by both Southern blotting and PCR methods identified the defective GPIIIa gene inherited by patients, and determined the genotype of asymptomatic subjects. Analysis of this Taq I polymorphism by PCR method should be potentially useful in future for the carrier detection and prenatal diagnosis in Glanzmann's thrombasthenia families.

China↗

Intestinal distension alters vagal efferent activity and small intestinal transport in vivo.

We investigated whether intestinal distension altered net water absorption by an adjacent noncontiguous segment of intestine and the mechanism for this effect. Influx and efflux catheters were placed in two intestinal loops, after which the bowel was transected between the loops. During perfusion with Ringers-HCO3-, the proximal loop efflux catheter was elevated 5, 10, or 15 cm above the plane of the rat. Net water absorption was measured in the nondistended caudal (control) loop. Elevation of the proximal loop efflux catheter caused distension and increased intraluminal pressure in this loop but did not alter the circumference or intraluminal pressure of the caudal loop. Distension increased net water absorption by the caudal loop if the vagi were intact and the splanchnic nerves transected. Afferent vagotomy prevented the distension effect. Using the intracellular recording technique, we were able to show that the increase in absorption resulted from a reduction in the activity of vagal neurons innervating the gastrointestinal tract. Intestinal distension reduced the neural activity of 24 of 26 vagal efferent neurons. Our results demonstrate that after splanchnectomy intestinal distension activates a neural circuit that includes afferent and efferent vagal fibers, resulting in increased water absorption.

Absorption↗

Management of major arterial injuries of the limbs in 166 cases.

The charts of 166 patients with major arterial injuries (175 arterial repairs) inflicted during periods of peace and war time were reviewed. One hundred and sixty-seven (95%) of the arteries repaired remained patent. Nine patients had limbs which developed ischemic contracture. Nine patients required amputations. Follow-up data was obtained from seventy-five patients with an average follow-up of five years and one month (six months to thirty years). Early diagnosis, prompt treatment including fasciotomies when indicated, complete debridement, appropriate coverage, meticulous surgical technique, and simultaneous treatment of concomitant injuries are all crucial factors in successful limb salvage. Major arterial injuries of the limb occur frequently in our patient population, especially in time of war. Successful treatment of major arterial injuries may be life-saving, as well as allowing limb salvage and restoration of limb function. The purpose of this study is to review our experience with repair of major arterial injuries.

Adolescent↗

Identification of the recombination site within the steroid 21-hydroxylase gene (CYP21) of the HLA-B47,DR7 haplotype.

The HLA haplotype A3-Cw6-B47-C4A91-BQ0-DR7 is associated with congenital adrenal hyperplasia (CAH), since it only carries the dysfunctional steroid 21-hydroxylase A pseudogene as well as the 5' adjacent complement C4A gene. The recombination site leading to the deletion of the complement C4B and steroid 21-hydroxylase B genes in this haplotype was studied by determining the 21-hydroxylase genomic DNA sequence in comparison to the standard CYP21A- and CYP21B-specific sequences. A 200-bp region between exons 7 and 8 was identified as a possible recombination site. Thus the deleted area comprises the 3' end of the CYP21A pseudogene, the entire C4B gene and the 5' end of the CYP21B gene. The findings were confirmed by PCR amplification of a 1.8-kb fragment of the CYP21 gene. This PCR system is specific for CYP21A/B recombinant genes and may be used for screening among CAH patients carrying this type of deletion.

Adrenal Hyperplasia, Congenital↗