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Biomedical subjects

X H Guo

Publications and source records attributed to X H Guo.

33 records · Page 2Linked to original sources
PubMed1996

Structure and sequence of the human sulphamidase gene.

Sanfilippo A syndrome (MPS-IIIA) is a mucopolysaccharide lysosomal storage disorder caused by a deficiency in the lysosomal enzyme, sulphamidase (EC 3.10.1.1), which is required for the degradation of heparan sulphate. A genomic clone containing the entire sulphamidase gene was isolated from a chromosome 17-specific gridded cosmid library. The structure of the gene and the sequence of the exon/intron boundaries and the 5' promoter region were determined. The sulphamidase gene is split into 8 exons spanning approximately 11 kb.

Base Sequence↗
PubMed1995

Expression, purification and characterization of recombinant human N-acetylgalactosamine-6-sulphatase.

Full-length cDNA sequences encoding human N-acetylgalactosamine-6-sulphatase were stably expressed in Chinese hamster ovary cells under the transcriptional control of the human polypeptide chain elongation factor 1 alpha gene promoter. A clonal cell line overexpressing recombinant N-acetylgalactosamine-6-sulphatase to a level of approx. 3 mg/l of culture medium was isolated. The secreted precursor enzyme was purified to homogeneity by a two-column procedure with an overall yield of 53% of the activity. The physical and catalytic parameters of the recombinant enzyme were similar to those of the mature form isolated from liver. On SDS/PAGE and gel filtration, recombinant N-acetylgalactosamine-6-sulphatase had a native molecular mass of 58-60 kDa. Recombinant N-acetylgalactosamine-6-sulphatase was endocytosed by mucopolysaccharidosis IVA fibroblasts via the mannose-6-phosphate receptor-mediated pathway and was efficiently localized to lysosomes.

Animals↗
PubMed1995

Cloning of the sulphamidase gene and identification of mutations in Sanfilippo A syndrome.

Sanfilippo A syndrome is one of four recognised Sanfilippo sub-types (A, B, C and D) that result from deficiencies of different enzymes involved in the lysosomal degradation of heparan sulphate; patients suffer from severe neurological disorders. The Sanfilippo syndrome sub-types are also known as mucopolysaccharidosis (MPS) type III (MPS-IIIA, B, C and D), and are part of the large group of lysosomal storage disorders. Each of the MPS-III types is inherited as an autosomal recessive disorder with considerable variation in severity of clinical phenotype. The incidence of Sanfilippo syndrome has been estimated at 1:24,000 in The Netherlands with MPS IIIA (MIM #252900) the most common. MPS-IIIA is the predominant MPS-III in the United Kingdom, and has a similar high incidence to that found in The Netherlands (E. Wraith, personal communication). There is a particularly high incidence of a clinically severe form of MPS-IIIA in the Cayman Islands with a carrier frequency of 0.1 (ref. 4). Due to the mild somatic disease compared to other MPS disorders there is difficulty in diagnosing mild cases of MPS-III, hence Sanfilippo syndrome may be underdiagnosed, especially in patients with mild mental retardation. Here, we report the isolation, sequence and expression of cDNA clones encoding the enzyme sulphamidase (EC 3.10.1.1). In addition, we report the chromosomal localisation of the sulphamidase gene as being 17q25.3. An 11-bp deletion, present in sulphamidase cDNA from two unrelated Sanfilippo A patients, is described.

Amino Acid Sequence↗
PubMed1995

Human renin 5'-flanking DNA to nucleotide-2750.

Renin is one of the most important factors in blood pressure and electrolyte regulation in mammals and the renin locus has been implicated in hypertension. To assist studies of promoter control we therefore determined the 5'-flanking sequence of the human gene (REN) to residue -2750 relative to the transcription start site (+1). Sites of homology to consensus sequences for binding of trans-acting factors involved in transcriptional control of other genes were identified, and functionality for two of these (a CRE and Pit-1 site) have so far been demonstrated.

Base Sequence↗
PubMed1994

Morquio A syndrome: cloning, sequence, and structure of the human N-acetylgalactosamine 6-sulfatase (GALNS) gene.

Deficiency of the lysosomal enzyme, N-acetylgalactosamine 6-sulfatase (GALNS;EC 3.1.6.4), results in the storage of the glycosaminoglycans, keratan sulfate and chondroitin 6-sulfate, which leads to the lysosomal storage disorder Morquio A syndrome. Four overlapping genomic clones derived from a chromosome 16-specific gridded cosmid library containing the entire GALNS gene were isolated. The structure of the gene and the sequence of the exon/intron boundaries and the 5' promoter region were determined. The GALNS gene is split into 14 exons spanning approximately 40 kb. The potential promoter for GALNS lacks a TATA box but contains GC box consensus sequences, consistent with its role as a housekeeping gene. The GALNS gene contains an Alu repeat in intron 5 and a VNTR-like sequence in intron 6.

Base Sequence↗
PubMed1994

[Autopsy analysis of 90 workers exposed to workplace dusts].

An analysis of 90 autopsy cases of workers exposed to workplace dusts showed that the older they were the higher the positive rate of pneumoconiosis and the more serious the pathological changes in the lungs. All the cases over 65 years of age were diagnosed with pneumoconiosis. The positive rate of pneumoconiosis diagnosed by pathology was associated with the duration of exposure to workplace dusts. The positive rate was 67.92% in cases exposed to workplace dusts for over 10 years and 93.3% in those exposed for over 30 years. 63.3% of all the autopsy cases reported here were given the diagnosis of pneumoconiosis after pathological examination, whereas only 13.3% were diagnosed clinically before they died. These data demonstrate the importance of autopsy in the diagnosis of pneumoconiosis.

Adult↗
PubMed1994

[Studies on culturing seeds of Tripterygium hypoglaucum (Level.) Hutch. in vitro].

This paper for the first time reports the embryo culturing in vitro of Tripterygium hypoglaucum. Tests were conducted of every condition for the normal germination and growth of the excised embryo. The result shows that the germination ratio reaches the highest on the white basic medium when the intensity of light is 400Lx, sucrose concentration is 3% and whole nitrogen content is 230 mg/L. Intact plantlets are obtained from the aseptic shoots. A method using aseptic shoots to accelerate the propagation of plantlets is found primarily.

Culture Media↗
PubMed1993

Sizing of large DNA molecules by hook formation in a loose matrix.

We present details on how to implement a newly developed methodology, Optical Contour Maximization (OCM), for accurately sizing large DNA molecules. Agarose gel containing stained DNA is cast between a slide and a coverslip, and molecules moved by an applied electrical field are observed using fluorescence microscopy. The molecules of interest lie in the interface formed between the coverslip and gel. DNA movement in this region is largely confined to lateral motions. When a DNA molecule snags an obstacle, it elongates, forming a metastable hook that can persist for several seconds. We found that the longest observed hook contour length can be determined from rapidly collected images. This maximized length shows a linear correlation with reported size [X.H. Guo, E.J. Huff & D.C. Schwartz, Nature 359, 783 (1992)] Successful measurements require a critical balance between the voltage needed for full elongation, and the unwanted effects of too large a voltage: fewer metastable hooks form, they dissipate faster, molecule breakage becomes a problem, and faster image collection becomes necessary. We measured apparent contour length as a function of applied voltage and determined an optimal voltage for our apparatus, using a singly anchored 114kb molecule. The measurement precision is estimated from the distribution of results. We expect that OCM will find utility in physical mapping and molecular karyotyping of lower eucaryotes of medical importance.

DNA↗
PubMed1993

The morquio A syndrome (mucopolysaccharidosis IVA) gene maps to 16q24.3.

The gene for N-acetylgalactosamine-6-sulfatase, the deficiency of which results in Morquio A syndrome (mucopolysaccharidosis type IVA), was assigned to chromosome 16 at band q24.3 by fluorescence in situ hybridization. Localization of this band was confirmed by PCR analysis of a somatic cell hybrid panel used for fine mapping of chromosome 16.

Animals↗
PubMed1992

Structure and sequence of the human alpha-L-iduronidase gene.

In humans, a deficiency of the lysosomal hydrolase alpha-L-iduronidase (IDUA;EC 3.2.1.76) results in the lysosomal storage of the glycosaminoglycans heparan sulfate and dermatan sulfate, thereby causing the lysosomal storage disorder mucopolysaccharidosis type I. The gene for IDUA is split into 14 exons spanning approximately 19 kb. We report the sequence of two non-contiguous segments of the IDUA gene, one 1.8-kb segment containing exons 1 and 2 and surrounding sequences and a second segment of 4.5 kb containing the last 12 exons. The potential promoter for IDUA has only GC box type consensus sequences consistent with a housekeeping promoter and is bounded by an Alu repeat sequence. The first two exons of IDUA are separated by an intron of 566 bp, then there is a large intron of approximately 13 kb, and the last 12 exons are clustered within 4.5 kb. No consensus polyadenylation signal was found in the 3' untranslated region, although two variant polyadenylation signals are proposed.

Base Sequence↗
PubMed1992

[Effects of Salvia miltiorrhiza Bunge on isolated perfused liver and portal vein of rats].

The experiment results showed that Salvia miltiorrhiza (SMB) had a protective effect on the isolated perfused liver injured by CCl4 of rats, made the activity of GPT in perfusate of SMB group lower than that of intoxicated groups and relieved the hepato-pathohistologic lesions of SMB group as compared to the intoxicated group. SMB had no significant effects on both the activities of SGPT in vitro and the portal vein of rats.

Alanine Transaminase↗
PubMed1992

[Obstructive sleep apnea syndrome and essential hypertension: diurnal variation of urinary catecholamines].

30 subjects of old and middle age (28 male, 2 female) with obstructive sleep apnea syndrome (OSAS) and 20 normal subjects with matchable age and body weight (14 male, 6 female) as control were studied with nocturnal polysomnography for at least 7 hours. Right arm blood pressure was determined in supine position before and after sleep. Meantime, three 8-hour urine specimens, two collected while awake and one during sleep were examined for urinary levels of epinephrine (E) and norepinephrine (NE) with fluorometric method. All OSAS subjects (mean apnea index 42.9) had significant arterial oxygen desaturation (mean 63.9%). 12/30 OSAS subjects had definit history of essential hypertension. They described that hypertension appeared months or years after the onset of sleep disorders. Before sleep the blood pressure in OSAS subjects was higher than that in controls (mean 133/90 mmHg versus 118/77 mmHg P < 0.001). After 7 hours of sleep with apnea events, the blood pressure rose to 149/100 mmHg (P < 0.001). whereas in the controls there was no change of statistic significance (mean 115/77 mmHg). A diurnal rhythm in free catecholamines excretion was apparent for both NE and E (P < 0.05) in the controls, while in OSAS there was no normal diurnal rhythm. 24-hour values of NE were remarkably higher than those in controls. It is known that up to 40% of OSAS subjects is in the population of essential hypertension.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1991

[Studies on the determination of the circulating antigen in experimental toxoplasmosis].

This paper reports on the regular pattern of growth and decline of the circulating antigen (CA) determined by I-ELISA in experimentally infected rabbits. Toxoplasma antigen was demonstratable in the circulation in 64.7% rabbits 1 day after tachyzoite inoculation and the level of the CA in sera from infected rabbits showed an increase on d4 and reached its peak level on d10-13 after inoculation but declined soon thereafter. CA was not detectable on days 60 to 90 after inoculation. The results suggest that determining CA is a useful means for diagnosing active toxoplasmosis and that I-ELISA might be a promising approach to detect CA in toxoplasmosis.

Animals↗
PubMed1990

Small-angle neutron scattering study of the structure of protein/detergent complexes.

Small-angle neutron scattering (SANS) was used to study the structure of protein/sodium dodecylsulfate complexes. Two water soluble proteins, bovine serum albumin (BSA) and ovalbumin (OVA), were used. The protein concentration was kept constant at 1 wt %, and protein/detergent wt ratio varied between 1/1, 1/1.5, 1/2 and 1/3. Absolute intensities of SANS distributions were analyzed by a fractal model. Analyses of large Q portions of SANS distributions established that sodium dodecylsulfate (SDS) molecules bound to a protein/SDS complex form micelle-like clusters. On the other hand, analyses of small Q portions of SANS distributions clearly showed that the arrangement of micelle-like clusters resembles a fractal packing of spheres. We showed that a protein/SDS complex can be characterized by four parameters extracted from the scattering experiment, namely, the average micelle size and its aggregation number, the fractal dimension characterizing the conformation of the micellar chains, the correlation length giving the extent of the unfolded polypeptide chains, and the numbers of micelle-like clusters in the complex.

Animals↗