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Biomedical subjects

G Xiao

Publications and source records attributed to G Xiao.

At least 91 records · Page 5Linked to original sources
PubMed1998

A preliminary clinical study of bifidobacteria preparation on the treatment of diarrhea in severely burned patients.

OBJECTIVE: To investigate the role of bifidobacteria preparation in the prevention of diarrhea in severely burned patients. METHODS: Forty-three severely burned patients who were inflicted by diarrhea were included in this study. The changes of intestinal microflora were observed. RESULTS: The intestinal flora of all patients with diarrhea changed greatly before treatment. The proportions of bifidobacteria and bacteroid decreased significantly, whereas those of aerobe and Candida increased relatively. The ratio of anaerobe to aerobe decreased. As a result, the patients' intestinal flora were distorted. After six days treatment of bifidobacteria, diarrhea in most patients ceased and the intestinal microflora restored. CONCLUSION: Bifidobacteria feeding plays an important role in restoring intestinal microflora and stopping diarrhea in severely burned patients.

Adolescent↗
PubMed1998

[Protection of hypertonic saline/mannitol to thymic cell apoptosis induced by endotoxin (LPS) in mice].

To observe the effect of HSM (hypertonic saline/mannitol) on thymic cell apoptosis induced by endotoxin (LPS) in mice, twenty-four mice were divided into three groups (8 mice/each group): 1. Control group, each mouse was intraperitonealy injected with normal saline; 2. LPS group, each mouse was intraperitonealy injected with LPS; 3. LPS + HSM group, each mouse was intraperitonealy injected with HSM before and after injection of LPS. The results showed that the percentage of apoptosis of HSM + LPS group was lower than that of LPS group (P < 0.01); the ladder of nucleosomal DNA fragments by agarose gel eletrophoresis of HSM group was not typical as compared with that of LPS group. It is suggested that HSM might protect the apoptosis of mouse thymic cell induced by LPS and may improve the immune function.

Adjuvants, Immunologic↗
PubMed1998

[Determination of the soluble non-starch polysaccharides in rice and wheat bran by gas chromatography].

Cereal polysaccharides can be broadly classified into two distinct and chemically well-defined types. They are the storage polysaccharides (alpha-glucan) and the structural polysaccharides starch (beta-glucan) which are usually called non-starch polysaccharides (NSP). The determination of soluble non-starch polysaccharides in rice and wheat bran by gas chromatography has been developed. The free sugars in the sample were extracted with 80% ethanol. The residue was hydrolyzed in an acetic acid buffer solution (pH 5.0) in the presence of amylase and amyloglucosidase to remove starch. The soluble NSP obtained was further hydrolyzed in the acidic condition to produce the corresponding monosaccharides which were derivatized to form alditol acetates for GC analysis with allose as the internal standard. The GC conditions were OV-1701 column (25 m x 0.3 mm) with temperature program from 195 degrees C to 225 degrees C and FID.

Chromatography, Gas↗
PubMed1997

Comparison of prostaglandin H synthase isoform structures using limited proteolytic digestion.

Prostaglandin H synthase (PGHS) catalyzes a key step in the biosynthesis of a variety of bioactive lipid mediators. The two known isoforms (PGHS-1 and -2) share about 60% amino acid identity, but exhibit distinct interactions with substrates, activators, and inhibitors. Ovine PGHS-1 has previously been shown to have a distinctive protease-sensitive site near Arg277; cleavage by trypsin, chymotrypsin, or proteinase K produces fragments of 33 and 38 kDa and loss of activity. The ovine PGHS-1 crystal structure shows Arg277 located in an exposed loop structure; homology modeling predicts similar loop structures for both human isoforms (hPGHS-1 and -2). We have used limited proteolytic digestion of recombinant hPGHS-1 and hPGHS-2 to probe their structures. Incubation of hPGHS-1 with either trypsin or proteinase K produced 33- and 38-kDa fragments and loss of activity. In contrast, incubation of hPGHS-2 with the same proteases led to cleavage of only a 2- to 3-kDa fragment, with no decrease in activity. Immunoblotting with site-specific antibodies demonstrated that the cleaved fragment originated from the hPGHS-2 C-terminus. Similar immunoblotting experiments indicated that trypsin did not attack the ovine PGHS-1 C-terminus. Mutagenesis was used to replace Pro263 of hPGHS-2 (corresponds to Arg277 of ovine PGHS-1) with arginine, inserting a potential trypsin site. Incubation of this P263R hPGHS-2 mutant with either trypsin or proteinase K resulted in cleavage near the C-terminus and retention of activity, just as with wild-type hPGHS-2. A peptide containing residues 259-268 of the P263R mutant was cleaved by trypsin at the same rate as a peptide corresponding to hPGHS-1 residues 272-281, demonstrating that the sequence differences were not responsible for the lack of tryptic cleavage at residue 263 in the hPGHS-2 mutant. Preincubation of hPGHS-2 with graded levels of guanidinium HCl before incubation with proteinase K did not produce large proteolytic fragments, indicating that the hPGHS-2 loop region was not selectively unfolding. The results point to two regions of significant structural difference between PGHS-1 and -2: the Arg277 loop, which is protease-sensitive in PGHS-1 but protease-resistant in PGHS-2, and the C-terminus, which is protease-sensitive in PGHS-2 but not in PGHS-1.

Amino Acid Sequence↗
PubMed1997

Antitumor activity of the novel human breast cancer growth inhibitor, mammary-derived growth inhibitor-related gene, MRG.

A novel human tumor growth inhibitor was identified by differential cDNA sequencing. The predicted amino acid sequence of this tumor-suppressing factor has a significant sequence homology to mouse mammary-derived growth inhibitor and thus was named mammary-derived growth inhibitor-related gene (MRG). MRG was found to be expressed in normal and benign human breast tissues but not in breast carcinomas. In situ hybridization analysis demonstrated a stage-specific MRG expression as follows. MRG was barely detectable in breast carcinomas, showed partial and weak expression in benign hyperplasia, but was expressed at a high level in normal breast epithelial cells. To determine if MRG can modulate in vivo growth of human breast cancers, we transfected a full-length MRG cDNA into MDA-MB-231 human breast cancer cells and studied the orthotopic growth of MRG transfectants versus control transfectants in the mammary fat pad of athymic nude mice. Overexpression of MRG in human breast cancer cells significantly suppressed cell proliferation in vitro and tumor growth in an orthotopic nude mouse model. These results suggest that MRG has tumor-suppressing activity, and the loss of MRG expression may be involved in the development and progression of breast cancer.

Amino Acid Sequence↗
PubMed1997

Analysis of hydroperoxide-induced tyrosyl radicals and lipoxygenase activity in aspirin-treated human prostaglandin H synthase-2.

A hydroperoxide-induced tyrosyl radical has been proposed as a key cyclooxygenase intermediate for the "basal" isoform of prostaglandin H synthase (PGHS-1). In the present study with the "inducible" isoform (PGHS-2), hydroperoxide was also found to generate a radical in high yield, a wide singlet at g = 2.0058 (29 G peak to trough). Reaction of PGHS-2 with a tyrosine-modifying reagent, tetranitromethane (TNM), resulted in cyclooxygenase inactivation and a much narrower radical EPR signal (22 G peak to trough). Addition of a cyclooxygenase inhibitor, nimesulide, similarly resulted in a narrow PGHS-2 radical. In PGHS-1, cyclooxygenase inhibition by tyrosine nitration with TNM or by active site ligands leads to generation of a narrow EPR instead of a wide EPR, with both signals originating from authentic tyrosyl radicals, indicating that the hydroperoxide-induced radicals in PGHS-2 are also tyrosyl radicals. Treatment of PGHS-2 with aspirin (acetyl salicylic acid, ASA) was previously shown to result in acetylation of a specific serine residue, cyclooxygenase inhibition, and increased lipoxygenase activity. Acetylation of PGHS-1 by ASA, in contrast, inhibited both lipoxygenase and cyclooxygenase activity. We now have found the ASA-treated PGHS-2 radical to be indistinguishable from that in control PGHS-2. Addition of nimesulide to ASA-treated PGHS-2 inhibited the lipoxygenase and resulted in a narrow radical EPR like that seen in PGHS-2 treated with TNM or nimesulide alone. Retention of PGHS-2 oxygenase activity was thus associated with retention of the native radical, and loss of activity was associated with alteration of the radical. Both native and ASA-treated PGHS-2 produced only the R stereoisomer of 11- and 15-HETE, demonstrating that the lipoxygenase stereochemistry was not changed by ASA. Native and ASA-treated PGHS-2 had lipoxygenase K(m) values considerably higher than that of the control PGHS-2 cyclooxygenase. Taken together, these results suggest that the same PGHS-2 tyrosyl radical serves as the oxidant for both cyclooxygenase and lipoxygenase catalysis and that acetylation of PGHS-2 by ASA favors arachidonate binding in an altered conformation which results in abstraction of the pro-R hydrogen from C13 and formation of 11(R)- and 15(R)-HETE.

Arachidonic Acid↗
PubMed1997

Identification of a breast cancer-specific gene, BCSG1, by direct differential cDNA sequencing.

A high-throughput direct-differential cDNA sequencing approach was employed to identify genes differentially expressed in normal breast as compared with breast cancer. Approximately 6000 expressed sequence tags (ESTs) from cDNA libraries of normal breast and breast carcinoma were selected randomly and subjected to EST-sequencing analysis. The relative expression levels of more than 2000 unique EST groups were quantitatively compared in normal versus cancerous breast. Of many putative differentially expressed genes, a breast cancer-specific gene, BCSGC1, which was expressed in high abundance in a breast cancer cDNA library but scarcely in a normal breast cDNA library, was identified as a putative breast cancer marker. In situ hybridization analysis demonstrated stage-specific BCSG1 expression as follows: BCSG1 was undetectable in normal or benign breast lesions, showed partial expression in ductal carcinoma in situ, but was expressed at an extremely high level in advanced infiltrating breast cancer. The predicted amino acid sequence of BCSG1 gene has a significant sequence homology to the non-amyloid beta protein fragment of the Alzheimer's disease amyloid protein. BCSG1 overexpression may indicate breast cancer malignant progression from benign breast or in situ carcinoma to the highly infiltrating carcinoma.

Amino Acid Sequence↗
PubMed1997

Ascorbic acid-dependent activation of the osteocalcin promoter in MC3T3-E1 preosteoblasts: requirement for collagen matrix synthesis and the presence of an intact OSE2 sequence.

Osteocalcin is a hormonally regulated calcium-binding protein made almost exclusively by osteoblasts. In normal cells, osteocalcin expression requires ascorbic acid (AA), an essential cofactor for osteoblast differentiation both in vivo and in vitro. To determine the mechanism of this regulation, subclones of MC3T3-E1 preosteoblasts were transiently transfected with 1.3 kb of the mouse osteocalcin gene 2 promoter driving expression of firefly luciferase. AA stimulated luciferase activity 20-fold after 4-5 days. This response was stereospecific to L-ascorbic acid and was only detected in MC3T3-E1 subclones showing strong AA induction of the endogenous osteocalcin gene. Similar results were also obtained in MC3T3-E1 cells stably transfected with the osteocalcin promoter. A specific inhibitor of collagen synthesis, 3,4-dehydroproline, blocked AA-dependent induction of promoter activity, indicating that regulation of the osteocalcin gene requires collagen matrix synthesis. Deletion analysis of the mOG2 promoter identified an essential region for AA responsiveness between -147 and -116 bp. This region contains a single copy of the previously described osteoblast-specific element, OSE2. Deletion and mutation of OSE2 in DNA transfection assays established the requirement for this element in the AA response. Furthermore, DNA-binding assays revealed that MC3T3-E1 cells contain OSF2, the nuclear factor binding to OSE2, and that binding of OSF2 to OSE2 is up-regulated by AA treatment. Taken collectively, our results indicate that an intact OSE2 sequence is required for the induction of osteocalcin expression by AA.

Animals↗
PubMed1997

[Respiratory monitoring and high frequence jet ventilation (HFJV) in the treatment of severe chest injuries].

Twenty-eight patients with severe chest injuries were classified into 3 groups: fractures of rib, hemopneumothorax and lung injured. Their respiratory rate (RR), SpO2 and blood gas were analysed. The results showed that SpO2 and PaO2 in lung-injured group were the lowest and RR was the highest in the 3 groups. After HFJV, there was a significant rise in PaO2 and SpO2 (P < 0.05) and a gradual recovery of respiratory and pulse rates in lung-injured group. The results suggest that respiratory monitoring is important for patients with chest injuries and HFJV may be used as an effective ventilation method for treating patients with severe chest injuries.

Adult↗
PubMed1997

[Analysis of triacylglycerol composition of perilla oil by combination of non-aqueous reversed-phase high performance liquid chromatography with gas chromatography].

Perilla oil was verified to be a rich source of polyunsaturated fatty acid (PUFA). Its linolenic content is 64.82%, the highest in the plants. Its triacylglycerol (TAG) components were isolated and identified first time in this paper by a combination of non-aqueous reversed-phase high performance liquid chromatography (RP-HPLC) with gas chromatography (GC). The TAG of perilla oil were isolated by HPLC with a Zorbax ODS column (5 microm, 4.6 mm i.d. x 250 mm; Dupont, Inc) and differential refractometer. The eluent was acetone/acetonitrile (80:20, V/V) with flow rate of 1.0 mL/min. The TAG component acyl groups were converted to fatty acid methyl esters (FAME). The acyl constituents for each TAG were determined by GC analysis. Gas chromatography of FAMEs was performed with a AC20 Carbowax 20M column (30 m x 0.32 mm i.d.). Detector and injection port temperatures were 260 degrees C. Column temperature was programmed from 120 degrees C, 1 min initially hold, then 8 degrees C/min to 220 degrees C and finally hold for 10 min. Nitrogen was the carrier gas. FAMEs peaks were identified by comparison of their retention times with those of standards. The retention times of the palmitic, stearic, oleic, linoleic, n-nonadecanoic acid (internal standard) and linolenic FAMEs were 10.7, 13.0, 13.3, 13.8, 14.2 and 14.7 min respectively. Five main components were determined. They were LnLnLn (34.10%), LnLnL (13.22%), LnLL (6.22%), LnLnO (11.38%) and LnLnP (9.17%). The TAG contents were corrected from their fatty acids content. The combination of non-aqueous RP-HPLC with GC is a simple and rapid method for the analysis of oil TAG structure and is also very helpful for the identification of oil quality.

Chromatography, Gas↗
PubMed1996

Identification and characterization of a 47 base pair activity-dependent enhancer of the rat nicotinic acetylcholine receptor delta-subunit promoter.

Nicotinic acetylcholine receptor (nAChR) genes are regulated by muscle electrical activity. E-box sequences found in their promoters are necessary for this regulation. However, many muscle genes contain E-boxes, yet are not regulated by muscle depolarization. This suggests that other elements are necessary, perhaps working in conjunction with E-boxes, to confer depolarization-dependent control onto promoter activity. We have used direct DNA injection into muscle as an in vivo assay to identify and characterize these additional elements. Mutagenesis and expression assays identified multiple elements within the first 81 base pairs (bp) of the nAChR delta-subunit promoter that contribute to its regulation by muscle electrical activity. Within this 81 bp sequence, two regions of DNA were identified that were capable of conferring activity-dependent regulation onto a heterologous promoter. The stronger of these two putative enhancers was characterized further. It is a 47 bp sequence that contains an E-box along with sequences similar to the SV40 core enhancer and an SP1 site. Site-directed mutagenesis identified residues within each of these sequences that were necessary for enhancer activity. Furthermore, methylation interference DNA footprinting assays showed increased nuclear protein binding to sequences within both these enhancers after muscle denervation, and this pattern of binding was very similar to that observed with nuclear protein isolated from myotube extracts. These latter results suggest that similar mechanisms may mediate increased nAChR expression during muscle development and after muscle denervation.

Animals↗
PubMed1996

First-sphere and second-sphere electrostatic effects in the active site of a class mu gluthathione transferase.

The activation of the thiol of glutathione (GSH) bound in the active site of the class mu glutathione transferase M1-1 from rat involves a hydrogen-bonding network that includes a direct (first-sphere) interaction between the hydroxyl group of Y6 and the sulfur of GSH and second-sphere interactions involving a hydrogen bond between the main-chain amide N-H of L12 and the hydroxyl group of Y6 and an on-face hydrogen bond between the hydroxyl group of T13 and the pi-electron cloud of Y6 (i.e., T13-OH---pi-Y6-OH--- -SG). The functions of these hydrogen bonds have been examined with a combination of site-specific mutagenesis and X-ray crystallography. The hydroxyl group of Y6 has a normal pKa of about 10 even though it is shielded from solvent and is in a largely hydrophobic environment. The apparent pKa of GSH in the binary Y6F.GSH complex is increased by 1.6 log units, and the reactivity of the enzyme-bound nucleophile is reduced. The catalytic properties of the Y6L mutant are identical to those of Y6F, suggesting that the weakly polar on-edge interaction between the aromatic ring and sulfur has no influence on catalysis. The refined three-dimensional structure of the Y6F mutant in complex with GSH shows no major structural perturbation of the protein other than a change in the coordination environment of the sulfur. Removal of the second-sphere influence of the on-face hydrogen bond between the hydroxyl groups T13 as in the T13V and T13A mutants elevates the pKa of enzyme-bound GSH by about 0.7 pKa units. Crystal structures of these mutants show that structural changes in the active site are minor and suggest that the changes in pKa of E.GSH are due to the presence or absence of the on-face hydrogen bond. The T13S mutant has a completely different side-chain hydrogen-bonding geometry than T13 in the native enzyme and catalytic properties similar to the T13A and T13V mutants consistent with the absence of an on-face hydrogen bond. The gamma-methyl group of T13 is essential in enforcing the on-face hydrogen bond geometry and preventing the hydroxyl group from forming more favorable conventional hydrogen bonds.

Animals↗
PubMed1996

A dual function activity-dependent, muscle-specific enhancer from rat nicotinic acetylcholine receptor delta-subunit gene.

Nicotinic acetylcholine receptors (nAChR) mediate communication between nerve and muscle. The expression of these receptors increases dramatically during muscle development when myoblasts are fusing into multinucleated myotubes. The molecular mechanisms mediating this muscle developmental stage specific expression are not well understood. We report here the identification of nAChR delta-subunit promoter DNA sequences that differentially interact with nuclear proteins isolated from myoblasts, myotubes, and nonmuscle cells. The functional role these sequences play in mediating muscle-specific expression was explored using mutagenesis and enhancer assays. These studies resulted in the identification of a 47-bp muscle-specific enhancer that mediates increased expression of the nAChR delta-subunit gene during myotube formation. This enhancer contains an E-box and an element with similarity to the SV40 core enhancer (SVCE). Point mutations throughout this 47-bp enhancer showed that the E-box and the SVCE sequence are both necessary for conferring muscle-specific expression onto a heterologous promoter. Interestingly, this same DNA sequence also functions as an activity-dependent enhancer.

3T3 Cells↗