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Biomedical subjects

W M Zhang

Publications and source records attributed to W M Zhang.

At least 55 records · Page 3Linked to original sources

Decreased affinity of K-receptor binding during reperfusion following ischaemic preconditioning in the rat heart.

The effects of ischaemic preconditioning with three cycles of ischaemia of 3 min and reperfusion of 5 min each cycle on ventricular fibrillation threshold (VFT) and ventricular fibrillation (VF), and binding properties of tritiated U69,593, a selective kappa opioid-receptor (k-receptor) agonist, during subsequent ischaemia and/or reperfusion were studied in the rat heart. It was found that ischaemic preconditioning significantly enhanced the VFT values during ischaemic and reperfusion. VF during the subsequent reperfusion period was also significantly reduced. The Kd of the [3H]U69,593 binding sites in the sarcolemma of the heart at 5 min of reperfusion was significantly increased following ischaemic preconditioning. The Bmax was, however, not altered after the preconditioning. The study provides evidence for the first time suggesting that the cardioprotective effects of ischaemic preconditioning may be related to a reduction in affinity of the K-receptor binding.

Analgesics↗

Complex formation between PSA isoenzymes and protease inhibitors.

PURPOSE: We have studied complex formation between the isoenzymes of prostate specific antigen (PSA) and protease inhibitors in vitro. MATERIALS AND METHODS: Ion exchange chromatography and hydrophobic interaction chromatography (HIC) were used for rapid separation of PSA isoenzymes from inhibitors and for characterization of the complex formation. Immunofluorometric assays (IFMA) specific for free PSA, the PSA-alpha 1-antichymotrypsin (PSA-ACT) complex and for both of these (total PSA) were used to measure various forms of PSA. Loss of free PSA immunoreactivity was used to estimate complex formation with alpha 2-macroglobulin (A2M) and ACT, which also was measured by PSA-ACT IFMA. RESULTS: Complex formation between PSA and A2M was more rapid than with ACT. After extended incubation, about 75% of PSA reacted with ACT and 85% with A2M. When added to a mixture of ACT and A2M at concentrations corresponding to those in plasma, only 17% of PSA formed a complex with ACT while 17% remained free and 66% was undetectable, indicating complex formation with A2M. After extended incubation of PSA-ACT at 37C, a significant proportion of PSA was released as free active PSA. When A2M was included in the reaction mixture, the loss of PSA-ACT was not accompanied by appearance of free PSA, an indication that it complexed with A2M. Five to 18% of nicked PSA complexed with ACT whereas 54 to 67% reacted with A2M. CONCLUSIONS: alpha 2-macroglobulin is the major inhibitor of PSA when it reached the circulation. Contrary to earlier assumptions, nicked PSA can bind to A2M rendering it inaccessible to antibodies.

Chromatography↗

[Study on the level of negative pressure of closed suction drainage of the peritoneal cavity].

An animal experiment and clinical investigations have been done to choose a suitable negative pressure of closed suction drainage of the peritoneal cavity. The results indicated that the efficiency of routine portable bulb suction devices giving 5.6-16 kPa was lower than vacuum bottles made by our hospital giving 58.86-73.15 kPa. This suggested that the level of negative pressure of the portable bulb suction devices can be increased to 58.86 to 73.15 kPa. The best one is 58.86 kPa.

Drainage↗

Multiplicity of kappa opioid receptor binding in the rat cardiac sarcolemma.

In order to determine the opioid receptor binding in the rat cardiac sarcolemma, direct and displacement binding assays of tritiated bremazocine, a nonselective opioid, agonist and U69593, a specific kappa-receptor agonist, were performed in a sarcolemmal membrane preparation. Saturation studies revealed that there were substantial binding sites for both radioligands. The [3H]bremazocine had a binding capacity of 100 fmol/mg protein with a Kd of 3.4 nM while the binding of [3H]U69593 showed a Kd of 7.4 nM and a Bmax of 139 fmol/mg protein. Displacement binding assay showed that [D-Ala2, MePhe4, Gly-(ol)5]enkephalin (DAGO), a specific mu-agonist, exhibited no inhibitory effect on the binding of either [3H]bremazocine or [3H]U69593. [D-Ala2,D-Leu5]enkephalin (DADLE), a specific delta-agonist, had no inhibition on the binding of [3H]U69593, but displaced the binding of [3H]bremazocine at higher concentrations (10(-5)-10(-6)M). U50488, a specific kappa-agonist, completed with the binding of [3H]U69593 23 times more potently than with the binding of [3H]bremazocine. A residual binding sites of [3H]bremazocine was found with a Kd of 6.4 nM and a Bmax of 76 fmol/mg protein under conditions in which mu, delta and kappa 1 opioid bindings were suppressed by DAGO and DADLE at 200 nM and U50488 at 1 microM. DADLE at 5 microM, a concentration known to block kappa 2 binding sites, abolished the residual U50488-insensitive specific binding of [3H]bremazocine. Met5-Enkephalin-Arg6-Phe7, a specific kappa 2 ligand, displaced the binding of [3H]bremazocine much more readily than the binding of [3H]U69593. The present study provided evidence for the presence of two kappa-receptor subtypes in the cardiac sarcolemma: one U50488-sensitive and DADLE-insensitive, and the other U50488-insensitive and DADLE-sensitive. These two kappa-receptor binding subtypes may correspond to kappa 1 and kappa 2 receptor subtypes. Computer assisted non-linear regression analysis in the competition experiments of U50488 with [3H]U69593 afforded a best fit of the inhibition curves for two displacing components of kappa 1 binding sites: one with high affinity and low Bmax, another with low affinity and high Bmax.

Adenosine Triphosphatases↗

Problems in the determination of prostate specific antigen.

Accurate determination of prostate specific antigen is important because of the increasing use of prostate specific antigen for diagnosis, follow-up and screening of prostate cancer. Standardization of this assay is complicated by the occurrence in serum of two major molecular forms of prostate specific antigen, free prostate specific antigen and a complex between prostate specific antigen and alpha 1-antichymotrypsin. These two forms of prostate specific antigen are recognized differently by many but not all antibodies. Thus, it is possible and desirable to develop methods that measure each form equally. To achieve better comparability, it is also necessary to prepare international standards for prostate specific antigen and its complex with alpha 1-antichymotrypsin. Furthermore, reference methods should be established which use these standards and carefully selected reference antibodies.

Humans↗

Standardization of PSA determinations.

Standardization of determinations for prostate specific antigen (PSA) has become an important issue due to the widespread use of these determinations for prostate cancer screening. Standardization of this assay is complicated due to the occurrence of two major forms of PSA in serum, the free antigen and a complex between PSA and alpha 1-antichymotrypsin (PSA-ACT). These two forms of PSA are recognized differently by different antibodies, but by careful selection of antibodies, it is possible to design methods that measure each form equally. It is suggested, that standards for PSA and PSA-ACT should be prepared and established as international standards. Furthermore, reference methods should be established on the basis that these standards and carefully selected reference antibodies.

Biomarkers, Tumor↗

Purification and characterization of different molecular forms of prostate-specific antigen in human seminal fluid.

We have developed a new procedure for purifying prostate-specific antigen (PSA) from human seminal fluid. The method is based on ammonium sulfate precipitation, hydrophobic interaction chromatography, gel filtration, and anion-exchange chromatography. It can be completed within 2 days with a recovery of intact PSA of 30%. By anion-exchange chromatography, five isoforms of PSA (A, B, C, D, and E) can be separated. The major form (PSA-B) consists of the intact enzyme, as shown by the occurrence of only one band of 33 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing or nonreducing conditions, and by amino acid sequencing, which reveals only one amino-terminal sequence corresponding to the reported amino-terminal sequence of intact PSA. The specific absorbance of 1 g/L PSA-B at 280 nm was 1.61, and 80% of the PSA-B formed a complex with alpha 1-antichymotrypsin, indicating that it is enzymatically active. Three cleaved forms of PSA with different nicking sites and low enzymatic activity were separated from intact PSA by ion-exchange chromatography. In addition, we isolated a glycosylation variant, PSA-A, which showed a higher isoelectric point (pI = 7.2) than PSA-B (pI = 6.9) but similar enzymatic activity; this form accounts for 5-10% of total PSA. After treatment with sialidase, PSA-A and B had the same isoelectric point value (pI = 7.7).

Amino Acid Sequence↗

Episodic growth hormone secretion in the grass carp, Ctenopharyngodon idellus (C. & V.).

The secretory pattern of growth hormone (GH) was studied in sexually immature grass carp (0.5-1.0 kg body wt). Serial blood samples were taken at 15-min intervals during 4- to 6-hr sampling periods via a dorsal aorta cannula. During each 4- to 6-hr sampling period plasma GH profiles demonstrated episodic secretion. Two patterns were observed, consisting either of one episodic of GH secretion (single or several pulses clustered) or two episodes of GH secretion during each sampling period. The interval between the two episodes of GH secretion was 2.6 hr. This is the first demonstration of episodic GH secretion in a teleost.

Animals↗

A simple device for sectioning fresh rat brain.

This article describes a simple device that enables sectioning of an unfixed unfrozen fresh rat brain corresponding in position with the de Groote's stereotaxic coordinate systems. The construction of this device allows adjustment of the thickness of the individual brain slices and, thus, facilitates quick and precise removal of whole discrete brain particles or nuclei from a minimal number of slices. Adaptation of this device for other stereotaxic brain atlases is easily possible.

Animals↗

Body fat and RNA content of the VMH cells in rats neonatally treated with monosodium glutamate.

The RNA content of the ventromedial hypothalamus (VMH), the lateral hypothalamic area (LHA), and the cortical neurons of male and female rats, neonatally treated with monosodium glutamate (MSG), were investigated. MSG (2 g/kg b.wt.) was injected subcutaneously to male and female rat pups daily for 5 days after birth. At 12 weeks of age a significant decrease of RNA content in the VMH cells and significantly increased body fat in neonatally MSG-treated animals were found. Correlation of these data showed a significant negative correlation between the body fat content and the RNA content in VMH neurons. The results a) confirm a closed relationship between the body fat content and the functional activity of VMH, b) indicate that obesity of neonatally MSG-treated animals should be due to decreased functional activity of the VMH cells.

Adipose Tissue↗

[Sequence analysis of N--ras and p53 gene mutation in the human lung adenocarcinoma cell lines].

Gene mutation and abnormal expression of ras oncogenes and p53 gene have a direct bearing on the carcinogenesis. Polymerase chain reaction (PCR), sequencing technique of ds DNA cycle and sequencing system were used to detect the gene mutation of N-ras as well as the exon 5 and 7 of p53 gene in hLA and LTEP-a2 cell lines of human lung adenocarcinoma. The result showed that mutation of both cell lines occurred on the 154th codon in exon5 of p53 gene, where GGC was displaced by GTC resulting a substitution of Val for Gly, nevertheless, N-ras oncogene and the exon7 of p53 gene are normal.

Adenocarcinoma↗

Body fat and RNA content of VMH neurons in rats.

The epididymal and retroperitoneal fat content of 180-day-old male Wistar rats, coming from litters of eight pups was correlated with their RNA content in ventromedial hypothalamus (VMH) and lateral hypothalamic area (LHA) neurons. A significant negative correlation was found between the RNA content in VMH neurons and epididymal and retroperitoneal fat content. The results indicate a closed relationship between the body fat content and the functional activity of VMH.

Adipose Tissue↗

Effects of discectomy on the stress distribution in the lumbar spine.

The change of the stress distribution of the lumbar spine after discectomy was analysed by the three-dimensional finite element method. It was shown that the stress level in the posterior element was higher, but that in the anterior element was lower than before disc excision. The most significant change of the stress distribution was found in the trabecular bone of the vertebral body. It is considered that in discectomy the normal disc tissue should be preserved as much as possible to maintain good function of the spine.

Biomechanical Phenomena↗

Influence of flexion-extension motion of lumbar spine on lumbosacral dural sac. An experimental study.

To analyze the effects of flexion-extension motion of lumbar spine on the position and shape of lumbosacral dural sac, 10 flexion/extension lateral myelograms of cadaver specimens of normal lumbar spine were measured. The results showed that moving from flexion to extension both sagittal diameter and length of the dural sac decreased significantly, the dural sac moved caudally, and the sagittal diameter of the anterior epidural space decreased in the mid-vertebral level but increased in the intervertebral level. The dynamic changes of the dural sac and their influence factors and clinical relevance were discussed.

Adult↗

Dynamic changes of the cervical ligamental flavum in hyperextension---hyperflexion movement and their measurements.

Ten fresh cadaver cervical spine specimens were frozen and bisected through the median sagittal plane. The length, thickness of the ligamental flavum (LF) and its protruding depth into the cervical canal were measured in hyperextension, hyperflexion and mid-position. The results showed that LF was elongated and became thinner in hyperflexion and shortened, thickened, and protruded into the cervical canal in hyperextension. The protruding depth into the canal was 3.5 +/- 0.35 mm at C 5-6, 3.25 +/- 0.50 mm at C 4-5 and 2.96 +/- 0.35 mm at C 6-7 respectively. Segmental stenosis was observed at the level of intervertebral space in hyperextension.

Adult↗

Effect of partial discectomy on the stability of the lumbar spine. A study of kinematics.

The effect on the stability of the lumbar motion segment, as affected by partial discectomy, was investigated using three-dimensional motion measurement technique. Eight functional spinal units from fresh cadaver lumbar spines were potted and clinically relevant loads applied through a special loading frame attached rigidly to the top vertebra of the specimen. Partial discectomy was performed on the right side of the specimen. The three-dimensional motions of each lumbar motion segment, before and after partial discectomy, were recorded. The results showed that partial discectomy significantly affects the stability of lumbar spinal motion segments. The clinical relevance of this finding is discussed.

Adult↗

Primary structure of rabbit skeletal muscle glycogen synthase deduced from cDNA clones.

The complete amino acid sequence of rabbit skeletal muscle glycogen synthase was deduced from cDNA clones with a composite length of 3317 bp. An mRNA of 3.6 kb was identified by Northern blot analysis of rabbit skeletal muscle RNA. The mRNA coded for a protein of 734 residues with a molecular weight of 83,480. The deduced NH2-terminal and COOH-terminal sequences corresponded to those reported for the purified protein, indicating the absence of any proteolytic processing. At the nucleotide level, the 5' untranslated and coding regions were 79 and 90% identical for rabbit and human muscle glycogen synthases, whereas the 3' untranslated regions were significantly less similar. The enzymes had 97% amino acid sequence identity. Interestingly, the NH2 and COOH termini of rabbit and human muscle glycogen synthase, the regions of phosphorylation, showed the greatest sequence variation (15 of 19 mismatches and two insertion/deletion events), which may indicate different evolutionary constraints in the regulatory and catalytic regions of the molecule.

Amino Acid Sequence↗

The effect of flexion-extension motion of the lumbar spine on the capacity of the spinal canal. An experimental study.

The symptoms of lumbar spinal stenosis are often posture-dependent, and it is generally accepted that the capacity of the spinal canal is affected by flexion-extension motions of the lumbar spine. A study of spinal capacity in flexion-extension of ten cadaver specimens on the lumbar spine was done and measurements were obtained from the flexion-extension lateral myelograms. There proved to be a larger capacity of dural sac of 3.5 to 6.0 ml (4.85 +/- 0.75 ml) in flexion than in extension, and the differences were highly significant (P less than 0.001). In addition, the sagittal diameter of the dural sac and the length of the spinal canal increased from extension to flexion. Because of the effect of flexion-extension motion of the lumbar spine on the spinal capacity, we suggest that maintaining the lumbar spine in flexion is more suitable since spinal canal capacity is enlarged and symptoms may be mitigated.

Adult↗