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Biomedical subjects

G Bai

Publications and source records attributed to G Bai.

At least 55 records · Page 3Linked to original sources

[The study of DSX-I type microcomputer autometic control injector system].

This paper studies the propulsive force of injector system controlled by 8031 single-chip microcomputer for Medical use By using one 8031 single-chip microcomputer as a microprocessor, the minimal autometic control system is constructed, which turns real-time control into reality. This product is a modern appliance used in diagnostic imaging medicine, in design of which the advanced instrument and electrical machinery are integrated into a unified structure. The clinic data demonstrate that the contrast medium can be injected to patients with rapidly and evenly. Dynamic CT Scanning can be intensified at any site of the body. x-ray dose exposed and lobour intensity to the operator can be decreased greatly, raise working efficiency.

Automation↗

Phenoloxidase activity in the reproductive system and egg masses of the pulmonate gastropod, Biomphalaria glabrata.

Phenoloxidase (PO) activity in the albumen gland (AG) and egg masses (EM) of Biomphalaria glabrata was assessed using high-performance liquid chromatography combined with electrochemical detection and colorimetric techniques. Both AG and EM extracts catalyzed the hydroxylation of L-tyrosine (monophenol oxidase activity, MPO) and oxidation of L-dopa (diphenol oxidase activity, DPO). However, no PO activity was found in the ovotestis. Both MPO and DPO activities in AG and EM were significantly inhibited by 1-phenyl-2-thiourea and inactivated by boiling. Approximately 35% of MPO and 44% of DPO activities were detected in the soluble fraction of homogenized EM, in contrast to that of homogenized AG, which contained about 5% and 12%, respectively, of MPO and DPO activities. N-acetyl-dopamine, a diphenolic compound, enhanced the hydroxylation of tyrosine by the PO. The presence of both MPO and DPO activities also was confirmed by the accelerated accumulation of dopachrome during incubation of EM extracts with L-tyrosine in the absence of ascorbate. Temperature and pH optima for this enzyme were 30 degrees C and 7.5, respectively. The potential roles of PO in egg formation in B glabrata are discussed.

Animals↗

Glucose metabolism and insulin release in mouse beta HC9 cells, as model for wild-type pancreatic beta-cells.

Glucose metabolism and its relationship with glucose-induced insulin release were studied in beta HC9 and beta TC3 cells to identify and characterize key factors controlling the intermediary metabolism of glucose and glucose-induced insulin release. The beta HC9 cell line, derived from pancreatic islets with beta-cell hyperplasia, is characterized by a normal concentration-dependency curve for glucose-stimulated insulin release, whereas the beta TC3 cell line, derived from pancreatic beta-cell tumors, shows a marked leftward shift of this curve. Maximum velocity and the Michaelis-Menten constant of glucose uptake in beta HC9 and beta TC3 cells were similar, even though GLUT-2 expression in these two cell lines differed. In both cell lines, the kinetic characteristics of glucose usage, glucose oxidation, and glucose-induced oxygen consumption were similar to those of glucose phosphorylation, indicating that the kinetics of glucose metabolism from the glucose phosphorylation step in the cytosol to the mitochondrial process of oxidative phosphorylation are determined by the glucose-phosphorylating enzyme, that is, by glucokinase in beta HC9 cells and by hexokinase in beta TC3 cells. Thus beta HC9 cells provide an opportunity for the quantitative analysis of glucose metabolism, the associated generation of coupling factors, and other essential beta-cell functions involved in glucose sensing and insulin secretion.

Animals↗

Specificities of five kinds of antisera produced against crude drugs, pinella tuber, hoelen, glycyrrhizae radix, trichosanthes root and panax ginseng.

Five kinds of rabbit antisera produced against five kinds of crude drugs, Pinella Tuber, Glycyrrhizae Radix (GR), Trichosanthes Root, Hoelen, and Panax Ginseng, and five kinds of extracts of the same crude drugs were prepared. Specificity of each antiserum was demonstrated by two immunological methods for analyses, the selected antibody enzyme immunoassay (SAEIA) and western blotting, and five crude drug extracts as specimens. Each crude drug extract contained characteristic antigen specific to the corresponding antiserum. Characteristic antigens were suggested as protein components. Characteristic antigen of the crude drug GR was separated from GR extract by three chromatographic procedures and a protein component was separated. The separated protein was successfully applied to develop a SAEIA method applicable for specific assays of both the separated protein and GR extract.

Animals↗

A new method for the detection and quantitative measurement of the contents of trichosanthes root component composing Chinese traditional medicines.

A new method was developed to estimate the content of Trichosanthes root (TR) component in two Chinese traditional medicines. Characteristic antigens of TR were separated from TR extract using rabbit antiserum specific for TR, a dried root tuber of Trichosanthes kirilowii. Two selected antibody enzyme immunoassay (SAEIA) methods, the SAEIA A for assay of TR extract and the SAEIA B for assay of karasurin A were used as detection methods for the separation of TR antigens. Using several column chromatographies, two kinds of TR antigens, a protein component and a glycan component, were separated from TR extract. A SAEIA C method for assay of TR glycan component was developed using biotinylated second antibody and peroxidase-labeled avidin as the detection method. The SAEIA C was also found applicable for specific assay of TR extract as well as for the contents of TR component present in two Chinese traditional medicines, prescriptions of which contained TR. Content of trichosantin, an abortifacient protein component of T. kirilowii, in both the medicines were also measured by applying the SAEIA B for assay of karasurin A. Little trichosantin was found in either medicine and the reason for this was determined.

Antibodies↗

[The therapeutic effect of somatostatin on acute severe pancreatitis: a clinical contrastive observation].

We observed the effect of somatostatin on the treatment of acute severe pancreatitis and on the inhibition of pancreatic secretion. 21 patients with acute severe pancreatitis were divided into control group (n = 12) and treatment group (n = 9) according to the admission time from 1992 to 1995. The control group was treated regularly and the treatment group was given intravenous somatostatin within 24h of onset 6mg/day for 5-7 days besides the regular treatment. No significant difference was noted in the general conditions of the two groups on admission. The volume of stomach suction in the somatostatin treated group was lower than that in the controls on the 2nd, 3rd and 4th admission days (P < 0.05). The serum amylase level of the treatment group was lower than that of the controls. 12 complications occurred in the somatostatin treated group as compared with 17 in the control group (P > 0.05). The clinical cure time was 15.6 +/- 4.8 days for the treatment group and 21.5 +/- 7.6 days for the controls (P = 0.02). We consider that as a pancreatic secretion inhibitor somatostatin can control the disease process and shorten the clinical cure time to some extent if it is used on the early stage of acute severe pancreatitis.

Adult↗

The maximum information principle and the evolution of nucleotide sequences.

The probability distributions of bases in nucleotide sequences are deduced from the maximum information principle by maximizing the entropy (due to random mutation of bases) under certain constraints (Markovian entropy, G + C content, etc, due to selection). Two formulations are given with respect to different selective constraints. The deviations of theoretical distributions from experimental data are lower than 10% for most sequences. It is shown that the Lagrange multipliers change from species to species systematically--i.e. selective constraints correlate with evolution.

Animals↗

Functional analysis of the proximal 5'-flanking region of the N-methyl-D-aspartate receptor subunit gene, NMDAR1.

The NMDAR1 receptor subunit is a common subunit of N-methyl-D-aspartate receptors. We have previously characterized 3 kilobases (kb) of 5'-flanking sequence of the NMDAR1 gene and now report on the ability of this region to direct transcription of a reporter gene and on its interaction with nuclear proteins. The sequence 356 base pairs (bp) 5' of the first nucleotide of codon 1 was sufficient to express a luciferase reporter gene in rat PC12 pheochromocytoma cells. Additional sequences upstream of nucleotide -356 influenced the activity approximately 2-fold. A labeled 112-bp fragment (position -356 to -245) formed six complexes (C1A and -B, C2A and -B, and C3A and -B), grouped as three double bands, with nuclear extracts from PC12 cells. Competition with Sp1 oligonucleotides abolished formation of C2A and -B and C3A and -B complexes. Sp1 antibody recognized the C3A complex in supershift experiments. Prior immunoprecipitation of nuclear extracts with Sp1 antibody abolished formation CA2 and -B and C3A and -B complexes. Purified Sp1 protein alone did not form a C3A complex but potentiated its formation when PC12 nuclear extract was added. A GC-rich sequence in this fragment was protected from DNase I digestion by nuclear extract. These results suggest that a 356-bp sequence comprises the NMDAR1 basal promoter, and that NMDAR1 gene expression may be regulated by Sp1-like nuclear factors.

Adrenal Gland Neoplasms↗

Spatial regulation of SpMTA metallothionein gene expression in sea urchin embryos by a regulatory cassette in intron 1.

The SpMTA metallothionein (MT) gene of the sea urchin Strongylocentrotus purpuratus is restricted in its expression to the aboral ectoderm in gastrulae and pluteus larvae. The proximal 1.6 kb of the 5'-flanking region together with the 1.12-kb first intron of the SpMTA gene are sufficient for its correct cell-type specific expression in transgenic embryos. This restricted spatial expression is largely eliminated by deletion of an interior 405-bp region in the intron. Within this region is a 295-bp, genomically repetitive, transposon-like segment (Nemer et al., 1993), containing several sequence motifs highly homologous to posited regulatory elements in the promoters of other genes (Thiebaud et al., 1990). The P3A and P5 sites in this apparent regulatory cassette were shown through competition to bind with relatively high affinities the same nuclear factors, bound by their counterpart sites in the CyIIIa actin promoter.

Actins↗

Determination of urinary acetylpolyamines by a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA).

A monoclonal antibody (mAb), ASPM-2, produced against N-(gamma-maleimidobutyryloxy)-succinimide (GMBS)-conjugated polyamine spermine [Spm; Fujiwara et al. (1994) Histochemistry 102, 397-404] was used for the development of an enzyme-linked immunosorbent assay (ELISA) for acetylpolyamines (Ac-PAs) in human urine. The ELISA is based on the principle of competition between an analyte and Spm-glutaraldehyde-bovine serum albumin conjugate-coated polystyrene microtiter wells for the mAb, followed by immunoreaction with biotinylated anti-mouse immunoglobulin and horseradish peroxidase-streptavidin. The ASPM-2 mAb showed strong immunoreaction with N1,N12-diacetylspermine (2Ac-Spm), N-monoacetylspermine (Ac-Spm), and N1-acetylspermidine (N1-Ac-Spd), the EC50 values being 29, 50, and 51 microM, respectively, but no cross-reaction with other PA-related compounds or amino acids. The method was used to measure urinary Ac-PA levels in healthy subjects and cancer patients, without pretreatment of the specimens, mean concentrations of 3.25 and 2.80 mumol per 24-h urine, respectively (as N1-Ac-Spd), being found. The ASPM-2 ELISA for N1-Ac-Spd, which is the PA most relevant to the analysis of human urine among the three Ac-PAs mentioned above, is specific and accurate, and can easily be used to analyze large numbers of specimens in parallel. It should thus have potential for studying the relationship between urinary N1-Ac-Spd levels and cancer.

Animals↗

c-fos promoter insensitivity to phorbol ester and possible role of protein kinase C in androgen-independent cancer cells.

In exploring the biological basis of androgen-independent prostate cancer, we observed serum-independent growth of androgen-independent cells. We then discovered that in androgen-independent but not androgen-dependent cells, the serum-responsive gene c-fos is insensitive to phorbol esters, which regulate c-fos through the same mechanisms as serum. Transient expression of protein kinase C, through which phorbol esters activate c-fos, was sufficient to desensitize c-fos in androgen-dependent cells. Incubation in protein kinase C inhibitor chelerythrine killed androgen-independent but not androgen-dependent cells. This finding implicates protein kinase C activity in androgen-independent prostate cancer and suggests novel therapeutic strategies.

Androgens↗

A mutant of protein phosphatase-1 that exhibits altered toxin sensitivity.

A chimeric mutant was constructed in which a 4-amino acid region (GEFD, residues 274-277) of rabbit muscle protein phosphatase-1 was replaced with the sequence YRCG corresponding to residues 267-270 of rabbit protein phosphatase-2A. This was based on the findings of a gene mutation in okadaic acid-resistant cells which results in a Cys-->Gly conversion in protein phosphatase-2A. The YRCG mutant of protein phosphatase-1 was expressed and purified. The properties of the mutant enzyme were investigated in terms of its sensitivity toward several toxin inhibitors (okadaic acid, microcystin, nodularin, calyculin A, and cantharidic acid), as well as inhibitor-2. The mutant enzyme exhibited a gain of function in the form of a 10-fold increased sensitivity toward okadaic acid that suggests this region is involved in toxin binding. Significant changes in sensitivity to inhibitor-2 and several of the other toxins were also observed, indicating that these may have a common binding region.

Amino Acid Sequence↗

Characterization of deletion mutants of the catalytic subunit of protein phosphatase-1.

Deletion mutagenesis was used to define the core region of the catalytic subunit of rabbit muscle protein phosphatase-1. Deletions in the N terminus were found to lead to loss of expression. Deletions of up to 33 residues from the C-terminal region were tolerated, and the truncated enzymes were fully active. Deletion of an additional 21 residues led to loss of expression. Mutants which had had 33 and 25 residues deleted maintained specific activities that were comparable to those of the wild type enzyme. The response of these two deletion mutants to okadaic acid, microcystin, and inhibitor-2 was determined. Only slightly lower IC50 values were observed in all cases, showing that the C terminus itself does not play a major role in the binding of these inhibitors. The deletion mutants formed stable complexes with inhibitor-2 as shown by gel filtration. These studies provide unambiguous evidence that the extreme C-terminal region of protein phosphatase-1 is not directly involved in catalytic function or in the binding of inhibitor-2, microcystin, or okadaic acid, and they also establish that the first approximately 300 residues of the sequence constitute a sufficient core for protein phosphatase-1 catalytic functions.

Amino Acid Sequence↗

Expression of recombinant inhibitor-2 in E. coli and its utilization for the affinity chromatography of protein phosphatase-1.

The coding sequence for rabbit muscle protein phosphatase inhibitor-2 was inserted into the pET3a expression vector. This vector allowed the expression of recombinant inhibitor-2 at levels of ca. 1.5% of the soluble protein. A simple procedure allowed the purification of inhibitor-2 from Escherichia coli lysates. This involved heat-treatment, followed by chromatography on Blue-Sepharose and Q-Sepharose. Recombinant inhibitor-2 inhibited rabbit muscle protein phosphatase-1 with a potency similar to that reported for the wild type protein. The recombinant protein was coupled to CH-Sepharose and this support was found to bind the catalytic subunit of protein phosphatase-1 with high efficiency. A procedure for a single-step affinity purification of recombinant ppase-1 from E. coli lysates was shown to be feasible.

Amino Acid Sequence↗

Primary structure of human liver glycogen synthase deduced by cDNA cloning.

The cDNA for human liver glycogen synthase was isolated by screening a human liver cDNA library constructed in lambda gt11. The full cDNA was 2912 bp in length. It coded for a protein of 703 amino acid residues with a molecular mass of 80.9 kDa. The number of amino acids was identical to and the deduced amino acid sequence homology was 92% that of the rat liver enzyme. The human and rat liver glycogen synthases are truncated by 34 amino acids compared to the human muscle enzyme, and by 32 amino acids compared to the rabbit muscle enzyme. The amino acid similarity between human liver and human muscle glycogen synthase was only 69%. It was least similar in the N and C terminal regions of the molecule. Two highly conserved regions are present in all published amino acid sequences for glycogen synthase, including those of the two yeast enzymes. These regions include the amino acid sequences from 201 to 400 and 501 to 600. This high conservation suggests that the catalytic site and the glucose-6-P and nucleotide allosteric sites are included in these regions.

Amino Acid Sequence↗

A clinical analysis of 38 patients with primary tracheo bronchial tumors.

During a 17-year period, 38 patients with primary tracheobronchial tumors received surgical treatment in PUMC Hospital. Among the 38, 12 had low-malignancy tracheal tumors, 11 had benign tracheal tumors, 14 had low malignancy bronchial tumors and 1 had a benign bronchial tumor. Fifteen operations were performed on 12 patients with low-malignancy tracheal tumors, including local resection of the tumor and tracheal wall in 4 and curettage of the tumor plus electrical cauterization in 10. Postoperative radiotherapy was used as an adjuvant treatment in 8 patients with adenocystic carcinoma. Eight patients have survived for more than 5 years and 3 patients have survived for longer than 10 years postoperatively. All 11 patients with benign tumors received curettage of the tumor and were followed up for an average of 6.5 years. Among them, 9 are still alive. Of the 14 patients with low malignancy bronchial tumors, 5 underwent curettage of the tumor plus electrical cauterization through incision of the main bronchus or intermedial bronchus, and 7 underwent lobectomy or pneumonectomy. The authors conclude that the correct diagnosis rate can be increased by enhancing recognition of this disease and applying tracheal tomography and bronchoscopy.

Adolescent↗

Highly identical cassettes of gene regulatory elements, genomically repetitive and present in RNA.

A region in the first intron of a metallothionein-encoding gene of the sea urchin Strongylocentrotus purpuratus (SpMTA gene) regulates its 5' promoter activity. Within this region is a 290-bp cassette of six sequence motifs that are present in other genes in this species and posited to operate as regulatory elements. The cassette, present at high multiplicity in the genome, was used to screen genomic DNA clones. Of these, six diverse individuals were partially sequenced and found to have segments 94% identical to the 290-bp cassette in the SpMTA gene. Their next 80 bp diverged from the SpMTA sequence but were highly identical among the six non-SpMTA clones and contained an additional regulatory motif. These diverse clones thus contained 370-bp cassettes of an overall 94% sequence identity and an apparent content of seven regulatory elements. The regulatory cassettes were transposon-like, insofar as the termini of the highly identical regions consisted of 24- to 25-bp inverted repeats, bracketed by 6- to 9-bp direct repeats in the divergent regions. In addition to being in transcripts of the SpMTA intron, the cassette was found in other sea urchin embryo poly(A)+ RNAs, in eggs and embryos, and enriched in pluteus ectoderm. The cassette sequence was present in moderate abundance in transcripts in both sense and antisense orientation. We report the presence of a transposon-like cassette of regulatory elements that is also represented in RNA, which potentially could function differently from previously described transposons.

Animals↗