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Biomedical subjects

D Liu

Publications and source records attributed to D Liu.

At least 721 records · Page 40Linked to original sources

HIV DNA and antibodies in syringes from injecting drug users: a comparison of detection techniques.

OBJECTIVE: Direct HIV testing of individual injecting drug users is not always feasible. As an alternative, we have evaluated the sensitivity and specificity of several techniques for detecting HIV-1-specific products in used syringes. DESIGN: Polymerase chain reaction (PCR) and antibody-capture assays were compared using syringes prepared with blood from HIV-1-positive and -negative individuals. METHODS: PCR sensitivity was maximized, enabling detection of single copies of HIV-1-specific proviral DNA. The limits of detection from used syringes were determined for PCR by diluting extracts and correlated to CD4+ cell counts. Similarly, limits of detection were determined for enzyme immunoassays (EIA) and Western blot. RESULTS: All techniques were highly specific, although with PCR false-positives were detected occasionally. EIA proved more sensitive than Western blot in detecting needles containing HIV-1-infected individuals' blood. Even after prolonged storage of syringes at room temperature, EIA was equal to or better than PCR as an HIV-1 detection technique. The most sensitive method for detecting HIV-1 was the viral-based EIA when the recommended predilution step was omitted. CONCLUSIONS: EIA proved preferable to PCR because of their higher sensitivity, absence of false-positives and easier sample preparation and analysis.

Blotting, Western↗

The toxic effects of serum from patients with type 1 diabetes mellitus on mouse neuroblastoma cells: a new mechanism for development of diabetic autonomic neuropathy.

The pathogenesis of diabetic neuropathy is incompletely understood. The possibility that humoral neurotoxic factors contribute as a cause of diabetic neuropathy was tested by application of serum from patients with Type 1 and Type 2 diabetes to mouse neuroblastoma cells, which have the characteristics of adrenergic neurons in culture. Serum from patients with Type 1 diabetes and somatic neuropathy significantly inhibited both proliferation and differentiation of neuroblastoma cells, while serum from patients with Type 1 diabetes but no symptoms of neuropathy and patients with Type 2 diabetes and neuropathy had no effect on proliferation, and serum from Type 2 patients only marginally inhibited differentiation. The effects of Type 1 diabetic serum could be reversed by pre-absorption of the serum to neuroblastoma cells, and were independent of glucose levels. Immunoglobulins precipitated from the sera mimicked the effects of whole sera. These results suggest that Type 1 diabetes mellitus causes a change in serum composition, possibly related to autoimmunity, that is capable of contributing to adrenergic autonomic neuropathy in diabetic patients.

Adult↗

An analysis of the glucagon response to hypoglycaemia in patients with type 1 diabetes and in healthy subjects.

The study aimed to analyse the glucagon response during hypoglycaemia in relation to gender, level of hypoglycaemia, and hyperinsulinaemia as well as its relation to other counterregulatory hormones in patients with Type 1 diabetes and in nondiabetic subjects. Mild hypoglycaemia was induced by an i.v. insulin infusion (244 pmol kg-1h-1) for 180 min in 43 Type 1 diabetic patients and 22 nondiabetic subjects. Venous blood glucose, plasma free insulin, glucagon, adrenaline, noradrenaline, growth hormone, and cortisol were measured every 15-30 min. The hormonal responses during hypoglycaemia were evaluated from the incremental areas under their respective curves. There was a linear correlation between the glucagon response and the decremental area of blood glucose (p < 0.005), but the slope of the regression line in the diabetic group was less steep than in the controls (p < 0.5), and, in spite of the deeper hypoglycaemia in the diabetic groups, their glucagon response was diminished (p < 0.05). Plasma, adrenaline, growth hormone and cortisol all increased during hypoglycaemia. The glucagon response correlated with the responses of growth hormone and cortisol in both groups, while it was positively correlated with the adrenaline response (p < 0.001) and inversely with the plasma insulin (p < 0.001) only in the diabetic patients. Although the insulin infusion rate was identical, the female diabetic patients had a lower metabolic clearance rate of insulin as compared with the males (p < 0.05). There was no statistical difference in the counterregulatory hormone responses between males and females in neither of the groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Modified polyvinyl alcohol-benzoporphyrin derivative conjugates as phototoxic agents.

Photosensitizing and biodistribution characteristics of a photosensitizer (benzoporphyrin derivative, monoacid ring A; BPD) conjugated to a macromolecule (modified polyvinyl alcohol; M-PVA, molecular weight = 10,000) were tested in vitro and in vivo. Modified PVA was loaded with BPD at molar ratios 1:12, 1:25, 1:50, 1:75 and 1:100. Most of the work was carried out with a conjugate having a 1:25 molar ratio. In vitro photosensitization was tested using A549 (human lung carcinoma), A432 (human epidermoid carcinoma) and P815 (mastocytoma of DBA/2 mice) cell lines. Photosensitization of M1 (rhabdomyosarcoma of DBA/2 mice) tumors was tested in an in vivo/in vitro assay, in which tumor-bearing mice were injected intravenously with free or conjugated 3H-BPD and 3 h later light activation of tumor cells was carried out in vitro. Biodistribution studies were carried out using M1 tumor-bearing DBA/2 mice and 3H-BPD either free or conjugated to M-PVA. The results of these studies showed that the conjugation of BPD to M-PVA resulted in the formation of a macromolecular photosensitizer that retained full photosensitizing activity of the photosensitizer molecules and at the same time gained new characteristics, advantageous for photodynamic treatment, especially in vivo. In vitro M-PVA-BPD conjugates were at least as efficient in photosensitization of tumor cells as an equivalent number of free BPD molecules, both in the presence and in the absence of serum. Although the biodistribution was in general comparable to free BPD, the conjugate (1:25) reached slightly higher levels in the blood, kidney, lung and spleen, and lower levels in the liver, brain, skin and muscle in comparison with free BPD.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Glycosyl transferases of O-antigen biosynthesis in Salmonella enterica: identification and characterization of transferase genes of groups B, C2, and E1.

In Salmonella enterica, there is a great variety of O antigens, each consisting of a short oligosaccharide (the repeating unit) repeated many times. The O antigens differ in their sugar composition and glycosidic linkages. The genetic determinants of the O antigen are located in an rfb gene cluster, and some, including those of S. enterica O serogroups B, C2, and E1, have been cloned and sequenced. In this study of the glycosyltransferases which form the glycosidic linkages, we identify and characterize the four mannosyl and three rhamnosyl transferase genes of the three rfb gene clusters.

Carbohydrate Sequence↗

Modulating effects of the extracellular sequence of the human insulinlike growth factor I receptor on its transforming and tumorigenic potential.

We reported previously that an N-terminally truncated insulinlike growth factor I receptor (IGFR) fused to avian sarcoma virus UR2 gag p19 had a greater transforming potential than did the native IGFR, but it failed to cause tumors in vivo. To investigate whether the 36 amino acids (aa) of the IGFR extracellular (EC) sequence in the gag-IGFR fusion protein encoded by the retrovirus UIGFR have a modulatory effect on the biological and biochemical properties of the protein, four mutants, NM1, NM2, NM3, and NM4 of the EC sequence were constructed. NM1 lacks the entire 36 aa residues; NM2 lacks the N-terminal 16 aa residues (aa 870 to 885), including two potential N-linked glycosylation sites of the EC sequence; NM3 contains a deletion of the C-terminal 20 aa residues (aa 886 to 905) of the EC sequence; and NM4 contains N-to-Q substitutions at both N-linked glycosylation sites. NM1 was the strongest of the four mutants in promoting anchorage-independent growth of transfected chicken embryo fibroblasts, while NM2 and NM4 had weaker transforming potential than did the original UIGFR virus. Only NM1 and NM3 were able to induce sarcomas in chickens. The four NM mutant-transformed cells expressed the expected proteins with comparable steady-state levels. The in vitro tyrosine kinase activity of P53NM1 was about fourfold higher than that of the parental P57-75UIGFR, whereas NM2 and NM4 proteins exhibited four- to fivefold-lower kinase activities. Despite lacking the IGFR EC sequence, P53NM1 formed covalent dimers similar to those formed by the parental P57-75UIGFR. Increased phosphatidylinositol (PI) 3-kinase activity was found to be associated with the mutant IGFR proteins. Among NM4 proteins. Elevated tyrosine phosphorylation of cellular proteins of 35, 120, 140, 160, and 170 kDa was detected in all mutant IGFR-transformed cells. We conclude that the EC 36-aa sequence of IGFR in the gag-IGFR fusion protein exerts intricate modulatory effects on the protein's transforming and tumorigenic potential. The 20 aa residues immediately upstream of the transmembrane domain have an inhibitory effect on the tumorigenic potential of gag-IGFR, whereas N-linked glycosylation within the EC sequence appears to have a positive effect on the transforming potential of UIGFR. Increased in vitro kinase activity and, to a lesser extent, in vivo tyrosine phosphorylation as well as the elevated association of PI 3-kinase activity with IGFR proteins seem to be correlated with the transforming potential of IGFR mutant proteins.

Animals↗

Distinctive effects of the carboxyl-terminal sequence of the insulin-like growth factor I receptor on its signaling functions.

We have shown previously that the extracellular sequences of the human insulin receptor (IR) and the insulin-like growth factor I receptor (IGFR) have an inhibitory effect on protein tyrosine kinase (PTK) activity and on the biological functions of their respective Gag-receptor fusion proteins. To study the role of IGFR carboxyl sequence in modulation of the Gag-IGFR PTK and biological activities, five mutants, CM1, CM2, CM3, CM4, and CM5, containing carboxyl deletions of 17, 27, 47, 67, and 88 amino acids (aa), respectively, were constructed from the parental virus UIGFR encoding the Gag-IGFR. Deletion of up to 27 aa had little effect on the cell-transforming and PTK activities of UIGFR. Deletions of 47 aa in CM3 abolished PTK and transforming activities. Surprisingly, a further deletion of 20 aa in CM4 beyond that in CM3 reactivated the kinase and transforming activities. CM5, containing a deletion of 20 aa beyond that in CM4, had only marginal transforming and PTK activities. We conclude that deletion of the carboxyl region of the Gag-IGFR inactivates, instead of activating as in the case with Gag-IR, its transforming activity and the amino acid sequence 1250 to 1310 is essential for PTK and transforming activities. Analysis of the ability of the full-length IGFR and its mutant receptors described above to associate with phosphatidylinositol 3 kinase indicated that the association required PTK activity and tyrosine phosphorylation of the receptors and correlated well with their transforming activities. The carboxyl 88 aa are not essential for the association.

Amino Acid Sequence↗

Adrenaline and noradrenaline responses during insulin-induced hypoglycaemia in man: should the hormone levels be measured in arterialized venous blood?

To compare arterial, arterialized venous and venous measurements of plasma catecholamines during hypoglycaemia, six healthy men were studied by an iv infusion of insulin (1034 pmol.kg-1.h-1). Arterial blood glucose was clamped at the baseline level for the first 30 min and then reduced to 3.2 and to 2.5 mmol/l for 20 min subsequently. At each stage, arterial, arterialized venous and venous blood samples were simultaneously taken for analyses of plasma catecholamines. Plasma glucagon significantly declined during hyperinsulinaemic euglycaemia and then increased during hypoglycaemia. Plasma adrenaline and noradrenaline increased significantly during hypoglycaemia. During euglycaemia and mild hypoglycaemia, arterial adrenaline tended to be higher and arterial noradrenaline lower as compared with the corresponding arterialized and venous values. During marked hypoglycaemia, the arterial catecholamines were significantly higher than that of the two venous samples (p < 0.05). There was a significant correlation between the arterial and the arterialized as well as venous catecholamine values (p < 0.005). We conclude that, during euglycaemia and mild hypoglycaemia, the arterialized adrenaline values are closer to the arterial levels whereas the arterialized noradrenaline concentrations are similar to those measured in venous blood. However, during marked hypoglycaemia, both the arterialized adrenaline and noradrenaline are 20-30% lower than the arterial values, being close to the venous levels.

Analysis of Variance↗

Surgical correction of upper eyelid retraction.

Over the last 7 years, I have used a "central aponeurosis disinsertion" technique to correct dysthyroid upper eyelid retraction in nine patients. Consistent and predictable results were achieved and maintained over follow-up periods ranging from 18 to 72 months. The technique is simple and effective, and does not involve extensive dissection or the use of a spacer.

Adult↗

The function of 3' noncoding sequences of hepatitis B surface antigen gene in a non-hepatic cell expression system.

A series of expression vectors containing the hepatitis B surface antigen (HBsAg) gene were constructed in which the promoter region of the HBsAg gene was replaced by the mouse metallothionein promoter. The HBsAg 3' noncoding sequences were modified by various deletions and/or additions of the heterogeneous genetic elements. The expression levels of the derivates were evaluated in a transient expression system using HeLa cells. We found that the enhancer I (EN-I) of hepatitis B virus (HBV) and the polyadenylation (poly A) signal are the most important elements for the expression of HBsAg gene in non-hepatic cells, in addition to the necessity to replace the original promoter of HBsAg gene. The heterogeneous poly A signal in combination with the simian virus (SV40) splicing signal could substitute for the poly A signal of the HBsAg gene, and, moreover, could increase the expression level of the HBsAg gene in HeLa cells. On the contrary, the enhancer II (EN-II) of HBV has no effect on the expression of the HBsAg gene in HeLa cells. These results imply another role of HBV EN-II on the stability of HBsAg transcripts in addition to the demonstrated function of activation the promoter.

DNA Mutational Analysis↗

[Changes of angiotensin II contents in rat plasma, brain, cardiovascular system and adrenal during stress].

The changes of content of angiotensin II (A II) in plasma, anterior hypothalamus, medulla oblongata, myocardium, vasculature and adrenals during acute and chronic stress were studied in rats. Compulsive cold-water swimming and trauma by limb-breaking were used to set acute stress models while cold environment of 4-8 degrees C was used to set chronic stress model. The results showed that: (1) The A II levels in plasma were significantly increased in all three stress- model groups, reaching to 900%, 390% and 134% of the control in the swimming group, the limb-broken group and the cold environment group, respectively. It's clear that the level of angiotension II in the acute stress groups were much higher than those in the chronic stress group. (2) The A II levels of the rat brain, myocardium and blood vessel were also increased in the stress animals except the limb-broken group. Furthermore, the contents of angiotensin II of the chronic stress animals were significantly higher than those of the acute stress animals in the brain, myocardium and blood vessels, but not in the adrenals. (3) The adrenal A II content was significantly higher than that of the control in all stressed animals. (4) The plasma corticosterone was also significantly increased over the control level in both the acute and chronic stress groups. These results suggest that circulating angiotensin II and tissue angiotensin II may play a role during the development of acute and chronic stress, respectively.

Adrenal Glands↗

[Effect of some factors on the development of exoerythrocytic stage of Plasmodium yoelii].

Using orthogonal design, the effect of environmental temperature, photoperiod, splenectomy and oestrogen levels upon the development of exoerythrocytic forms (EEF) of P. yoelii was observed. The results indicated that a significant number of viable sporozoites were cleared in the spleen, since the EEF density was much higher in the livers of splenectomized rats (1.73/mm3) than in sham-operated counterparts (0.55/mm3). The effect of high level oestrogen on EEF density was also evident since there was a significant difference between experimental (0.86) and control rats (1.42). Low environmental temperature caused the development of EEF stunted and asynchronous, but no significant effect on the density of EEF in this group was found. The density and average diameter (AD) of EEF between rats illuminated 8 and 16 hours per day were identical. The interaction between splenectomy and elevated oestrogen level offset each other, whereas the interactions between other two factors showed no difference by analysis of variance.

Animals↗

[Isolation and identification of gancaonin P-3'-methylether from the leaves of Glycyrrhiza uralensis Fisch].

Two flavonoids were isolated from the leaves of Glycyrrhiza uralensis Fisch (Licorice, Leguminosae). On the basis of physico-chemical properties and spectroscopy (UV, 1HNMR and MS), a new compound was elucidated as 3,5,7,4'-tetrahydroxy-3'-methoxy-6-isoprenyl flavone (gancaonin P-3'-methylether) and another known compound was identified as 8-C-prenyleriodictyol.

Drugs, Chinese Herbal↗

[Two new isoprenyl flavonoids from the leaves of Glycyrrhiza uralensis Fisch].

In previous papers, we reported flavonoids isolated from the leaves of Glycyrrhiza uralensis on the west of Inner Mongolia. Three of these flavonoids had the isoprenyl on the B ring, one was phenolic glucoside and six were flavonoidic glucosides. In the present paper, we report the isolation and identification of flavonoids from the leaves of this plant. The structures of these compounds have been identified as 5,7,3',4'-tetrahydroxy-3-methoxy-5'- isoprenylflavone(I),5,6,3',4'-tetrahydroxy-3-methoxy-6'-isopren ylfavone(II) and quercetin (III) by chemical method and spectroscopic analyses. I and II are new compounds and named uralenol-3-methylether and uralene, respectively. III was found for the first time in this species.

Flavonoids↗

A functional tomato ACC synthase expressed in Escherichia coli demonstrates suicidal inactivation by its substrate S-adenosylmethionine.

1-Aminocyclopropane-1-carboxylate (ACC) synthase is a key enzyme in the biosynthesis of the plant hormone, ethylene. We have isolated, sequenced and expressed a functional tomato (cv Pik-Red) ACC synthase gene in Escherichia coli. ACC synthase expressed in E. coli was inactivated by incubation with S-adenosylmethionine (SAM), the half-time of which was concentration dependent. Mixing the tomato fruit protein extract with the cell-free extract from transformed E. coli did not affect SAM-dependent inactivation of ACC synthase activity. Thus, single isoforms of the ACC synthase enzyme, which demonstrate the biochemical features expected of the tomato fruit enzyme, can be expressed in E. coli and their structure-function relationships investigated.

Amino Acid Sequence↗