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Biomedical subjects

H C Zhang

Publications and source records attributed to H C Zhang.

34 records · Page 2Linked to original sources

[Tear malate dehydrogenase, lactate dehydrogenase, and their isoenzymes in normal Chinese subjects and patients of ocular surface disorders].

Levels of malate dehydrogenase (MDH), lactate dehydrogenase (LDH) and their isoenzymes in tears of normal Chinese subjects and patients with ocular surface disorders were determined. The normal values of tear LDH and MDH were found to be 45.51 +/- 23.00-81.35 +/- 37.84 mumol.s-1/L and 11.00 +/- 5.33-19.50 +/- 9.17 mumol.s-1/L respectively, disregarding sex or eye distinction. The LDH/MDH ratio reflected sensitively the metabolism of corneal and conjunctival epithelium. The MDH isoenzymes comprised MDHs and MDHm, the former accounting for 80.0%-89.1%. The LDH isoenzymes comprised 5 varieties, of which the ratio H/M of subunit H to subunit M was 0.196 +/- 0.02. The changes in LDH isoenzymes were helpful to the differential diagnosis of external eye diseases, and the increase of MDHm reflected sensitively the degree of injury to the corneal epithelium.

Adolescent↗

[Keratitis associated with contact lens wear].

Keratitis associated with contact lens wear in 50 eyes (37 patients) was analyzed. The results revealed that lenses of extended-wear induced more cases of keratitis than did lenses of daily-wear. The pathogenetic mechanism was discussed.

Adult↗

Purification and characterization of recombinant mouse thymidylate synthase.

Recombinant mouse thymidylate synthase (TS) expressed at high levels in Escherichia coli was purified to homogeneity in greater than 70% yield by a rapid three-step procedure. Both 0.1% Triton X-100 and 10% glycerol were required to stabilize the enzyme whose activity remained unchanged after 1 month when stored at -20 degrees C. Thermal inactivation of the enzyme was a first-order process at 37 degrees C, with t1/2 values of 6.9, 15.6 and 3.0 min at pH 5.5, 7.0 and 8.5, respectively. The presence of saturating levels of dUMP at pH 8.5 increased the t1/2 of inactivation of 38 min. The pH profile for enzyme activity showed a narrow optimum region centered at pH 7.0, which was mirrored by the shape of the Km, dUMP/Vmax plot. The pH dependence of Kd for the covalent inhibitory ternary complex of enzyme, 5-fluoro-2'-deoxyuridylate and 5,10-methylenetetrahydrofolate exhibited a broad minimum between pH 5.5 and 8.5, and ranged between 3.1, 0.8 and 1.1 nM at pH 5.5, 7.0 and 8.5, respectively. The UV/VIS spectrum of the native enzyme exhibited a maximum at 280 nm (epsilon = 98,200 M-1 cm-1), while that of the inhibitory ternary complex showed an additional maximum at 320 nm. The 19F-NMR spectrum of the mouse enzyme:FdUMP binary complex revealed two new resonances at -2.8 and -34.8 ppm. The most deshielded resonance represented the noncovalent binary complex while the other resonance was assigned to the nucleotide covalently bound to the enzyme. The alteration of nucleotide binding equilibria produced by addition of H4 folate was exemplified by both an increase in intensity and a 5 ppm deshielding of the resonance attributed to the covalent FdUMP-enzyme complex. Addition of formaldehyde to the latter mixture produced the covalent ternary complex which resulted in the collapse of the resonances at -2.8 and -39.5 ppm and the appearance of a new resonance at -12.4 ppm.

Animals↗

[5-Fu as remedy for unsuccessful glaucoma filtration surgery].

Subconjunctival 5-Fu injections were administered to 10 glaucomatous eyes (8 patients) with unsuccessful previous filtration surgery, resulting in improvement of the filtration bleb and restoration of normal intraocular pressure to 9 eyes. Corneal exfoliation occurred in 2 eyes, which healed after cessation of the injections.

Adolescent↗

Site-directed mutagenesis of mouse thymidylate synthase: alteration of Arg44 to Val44 in a conserved loop guarding the active site has striking effects on catalysis and nucleotide binding.

The arginine located at position 44 of mouse thymidylate synthase is in a highly conserved loop that is in close proximity to the active site cleft of the enzyme. Structural analyses have suggested that this arginine forms hydrogen bonds with the alpha-carboxylate of the C-terminal amino acid and the phosphate of the substrate analog, FdUMP (D. A. Matthews, K. Appelt and S. J. Oatley, (1989) Adv. Enz. Reg., 29, 47-60). We have used protein engineering techniques to change this amino acid residue to valine. This alteration leads to large reductions in the ability of the enzyme to form covalent complexes with substrate (dUMP) or inhibitor (FdUMP) and at least a 100-fold reduction in catalytic activity. These observations show that this arginine plays an important role in maintaining catalytic activity of the enzyme.

Amino Acid Sequence↗

[An investigation of vital staining of the ocular surface and BUT of the tear film in Chinese].

Vital staining of the ocular surface with fluorescein and rose bengal and the tear film breakup time were studied in 268 healthy Chinese with the following results: Fluorescein did not stain the conjunctiva while it stained the cornea increasingly with age after 31 years. Rose bengal did not usually stain the cornea before the age of 30, the lacrimal caruncle and the semilunar fold before the age of 10, while it stained the bulbar conjunctiva of any age. BUT was 11-50 sec for 68.9% of the 268 subjects and the percentage less than or equal to 10 sec was 22.6%, which increased with age. The author concluded that rose bengal staining and BUT determination reflected the aging changes of the ocular surface and lacrimal secretion; however, for the Chinese, Rb staining of bulbar conjunctiva and BUT less than or equal to 10 sec alone should not be regarded as morbid.

Adolescent↗

Efficient synthesis of mouse thymidylate synthase in Escherichia coli.

The coding region of the mouse thymidylate synthase (TS)-encoding cDNA (ts) was inserted downstream from the phage T7 promoter and translation initiation signals of the expression vector, pET-3a, and transformed into Escherichia coli BL21(DE3)[pLysS]. When the wild-type (wt) cDNA sequence was used, mouse TS was synthesized in the bacterial cells in response to induction, but the level of expression was low. When the second codon (Leu) was changed from CUG, found in the normal mRNA, to CUU, the level of expression increased 17-fold and TS represented 5-10% of total cell protein. The recombinant enzyme was purified to homogeneity by affinity chromatography. The recombinant TS had the same Mr as the enzyme from cultured mouse fibroblasts. Kinetic studies with the recombinant enzyme showed that the apparent Km values for deoxyuridylate and 5,10-methylenetetrahydrofolate were 10.5 and 22 microM, respectively, which were similar to the values for TS from mouse cell extracts. The mouse ts expression vector will be useful for the large-scale production of the wt enzyme and for the creation and analysis of mutant enzymes by protein engineering techniques.

Animals↗

Cryptophthalmos: a report on three sibling cases.

Three siblings, two females and one male, with the rare disease, cryptophthalmos, are reported on. Cryptophthalmos is usually familiar, inherited as an autosomal recessive trait. The clinical manifestations indicate that the eyeball is present but disorganised; the lid muscles, extraocular muscles, and the nerve elements controlling ocular movements in the brain are developed. Treatment is of no avail, but in unilateral cases the partial lid coloboma, anterior symblepharon, and corneal opacification of the opposite eye may be somewhat improved by surgery.

Abnormalities, Multiple↗

Cognitive styles: illusory phenomena.

The relationship between cognitive style and illusion magnitude was examined using the assimilation and contrast Delboeuf distortions. As in previous research, field dependence correlated positively with assimilation; however, a significant negative correlation with contrast was also found, suggesting that, while field dependence may involve the illusory integration of the stimulus field, field independence may involve its equally illusory differentiation.

Adolescent↗

Phosphorus losses to water from lowland rice fields under rice-wheat double cropping system in the Tai Lake region.

To assess P losses to surface water by runoff during the rice season and by drainage flow during the winter wheat season, serial field trials were conducted in different types of paddy soils in the Tai Lake Region (TLR) during 2000 and 2001. Four P application rates were set as 0 (CK), 30, 150, and 300 kg P/hm2 for flooded rice trials and 0 (CK), 20, 80, 160 kg P/hm2 for winter wheat trials respectively. Field experiments were done in two locations with a plot size of 30 m2 and four replications in a randomized complete block design. A simplified lysimeter was installed for each plot to collect all the runoff or drainage flow from each event. Total P (TP) losses to surface water during rice season by runoff flow from four treatments were 150 (CK), 220 (T30), 395 (T150), 670 (T300) g P/ hm2 in year 2000, and 298, 440, 1828, 3744 g P/hm2 in year 2001 respectively in Wuxi station, here the soil is permeable paddy soil derived from loam clay deposit. While the losses were 102, 140, 210, 270 in year 2000, and 128, 165, 359, 589 g P/hm2 in year 2001 respectively in Changshu station, here the soil is waterlogged paddy soil derived from silt loam deposit. During the winter wheat season, total P lost from the fields by drainage flow in the four treatments were 253 (CK), 382 (T20), 580 (T89), 818 (T160) g P/hm2 in year 2000--2001, and 573.3, 709.4, 1123.2, 1552.4 g P/hm2 in year 2001--2002 at the Wuxi station. While these were 395.6, 539.1, 1356.8, 1972.1 g P/hm2 in year 2000--2001, and 811.5, 1184.6, 3001.2, 5333.1 g P/hm2 in year 2001--2002 at the Changshu station. Results revealed that P fertilizer application rates significantly affected the TP concentrations and TP loads in runoff during the rice season, and by drainage flow during the winter wheat season. Both TP loads were significantly increased as the P application rate increases. The data indicate that TP losses to surface water were much higher during the winter wheat season than during the rice season in two tested sites. The data also reveal that the annual precipitation and evaporation rate affected the soil P losses to surface water significantly. Year 2000 was relatively dried with higher evaporation thus P losses to water by both runoff and drainage flow were less than in year 2001 which was a relatively wet year with lower evaporation. Results indicate that texture, structure of the soil profile, and field construction (with or without ridge and deep drains) affected soil P losses to surface water dramatically. Annual possible TP lost to water at the application rate of 50 kg P/hm2 year tested in TLR were estimated from 97 to 185 tones P from permeable paddy soils and 109-218 tones P from waterlogged paddy soils. There was no significant difference of TP lost between the CK and the T50 treatments in both stations, which indicate that there is no more TP lost in field of normal P fertilizer application rate than in control field of no P fertilized. Much higher TP lost in runoff or drainage flow from those other P application rates treatments than from the T50 treatment, which suggest that P losses to surface water would be greatly increasing in the time when higher available P accumulation in plough layer soil in this region.

Agriculture↗