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Biomedical subjects

M Xu

Publications and source records attributed to M Xu.

At least 307 records · Page 17Linked to original sources

[Apolipoprotein A-IV and transportation of cholesterol from smooth muscle cells in experimental hyperlipidemia].

The effects of apolipoprotein (apo) A-IV and apoA-IV/phosphatidylcholine (PC) liposome on cholesterol efflux from the smooth muscle cells originating from the aorta of hypercholesterolemic rabbit model and control rabbits, and on the activation of lecithin cholesterol acyltransferase (LCAT) were studied respectively. Both apoA-IV/PC and apoA-I/PC liposomes have similar efficiency as the HDL's, i.e. a strong ability to clear intracellular cholesterol and the ability to activate LCAT. There were no differences noticed on the clearance ability between apoA-IV and apoA-I or between apoA-IV/PC and apoA-I/PC liposomes. These results suggest that apoA-IV may play an important role in the process of reverse cholesterol transport and apoA-IV/PC liposome may be effectively used instead of natural HDL to prevent the development of atherosclerosis.

Animals↗

PDI-, PPI- and chaperone-catalyzed refolding of recombinant human IL-2 and GM-CSF.

The studies on PDI-, PPI- and chaperone-catalyzed refolding of recombinant human IL-2 and GM-CSF show that PDI can prevent the mismatch of disulfide bonds and formation of aggregates by interchains linkage; furthermore, PDI can correct the mismatching of disulfide bonds in IL-2 isomers. PPI can increase the rate of folding reaction while chaperone can prevent the aggregation during the folding process. In addition, there is a synergistic effect between them.

Amino Acid Isomerases↗

Progress in surgical treatment of hydatid diseases in China. A clinical analysis of 22,005 surgical cases.

Altogether 27,716 cystic hydatid cases and 445 alveolar hydatid cases clinically diagnosed and surgically treated in China were retrospectively studied. In accordance with the parasitic sites of the hydatid cysts and the various pathological types, different surgical procedures were adopted. In an attempt to prevent intraoperative accidents and postoperative complications, continued improvements in operational techniques have been made. Different surgical procedures were evaluated for their advantages and disadvantages as well as their indications. According to the pathological changes of the residual cavities of ectocysts, different measures were adopted. In screening operations, injection of antihydatid drug into hydatid cysts followed by oral administration of this drug was used to prevent relapses. The therapeutic effect of all these measures has been proved satisfactory by follow-up studies. This indicates that the progress in surgical treatment of cystic hydatid disease during the recent 10 years has been attaining to the objectives of shortening the duration of treatment, decreasing the postoperative recurrence rate and increasing the cure rate. In some hospitals, no operative death and relapse of cyst growth have been reported in nearly 1,000 surgical cases of cystic hydatid disease. Only 10.8% of the 445 alveolar hydatid cases underwent radical operations and the prognoses were unfavourable. This was due to the absence of symptoms in early stages of the disease which often made the patients miss the chance for radical operation in hospitals.

Echinococcosis↗

[Studies on mode of human infection with Echinochasmus liliputanus].

Epidemiological survey and experimental studies on the mode of human infection with Echinochasmus liliputanus were carried out in Chenqiaozhou Village, Hexian County, Anhui Province in 1992. The results showed that humans could be infected through drinking unboiled water containing E.liliputanus cercaria or eating raw fish containing the metacercaria. The infection rates of E. liliputanus were 1.5% in the inhabitants who did not drink raw water, and 20.1% in those who used to drink raw water. It was only the cercaria, not the metacercaria that was detected in the water from ponds of this village. None of the investigated inhabitants ate uncooked fish. The results indicated that drinking raw water containing E.liliputanus cercaria was a major way through which a humans were infected and eating food containing the metacercaria was a minor mode of human infection.

Animals↗

[Synthesis and positive inotropic activity of 3,4-dimethoxybenzoyl-piperazine derivatives].

We designed and synthesized twelve new 3,4-dimethoxybenzoyl-piperazine derivatives. These chemical structures (TM-1-TM-12) were confirmed by means of IR, MS, H-NMR and elemental analysis. Compounds (OPC-8212 TM-1, TM-2, VPA) were tested for positive inotropic activity. The results showed that compounds OPC-8212 and VPA possessed positive inotropic activity (P < 0.05). Further pharmacological studies will be made.

Animals↗

Purification of recombinant human granulocyte-macrophage colony stimulating factor expressed in Escherichia coli.

Recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) was expressed as inclusion bodies (IB) in E. coli. A simple and effective protocol has been worked out for the purification. IB collected after the breakage of bacteria through sonication were subjected to repeated washing followed by solubilization in TE buffer (50 mmol/L of Tris.HCl, 1 mmol/L of EDTA, pH 8.3) containing 8 mol/L urea and 10 mmol/L DL-dithiothreitol. By means of Sephacryl-200 HR, refolding, and Q Sepharose Fast Flow, rhGM-CSF was obtained with a purity of 99%. The total protein recovery was 10% and specific activity of rhGM-CSF was 1 x 10(7) u/mg. The sequence of N-terminal 16 amino acid residues of purified rhGM-CSF was determined and found to be identical to the native protein. This study provided useful parameters for mass production of rhGM-CSF.

Amino Acid Sequence↗

[High efficiency expression of IL-4 using modified cDNA expression vector in mammalian cells].

In order to express eukaryotic gene in eukaryotic cells, a cDNA library was constructed and screened with synthesized probe of IL-4 cDNA. The obtained cDNA fragment 739 bp in length was inserted into pi H3M plasmid and expressed (8 x 10(3) IU/ml) in COS cells. A pdB-HhIL-4 expression plasmid was constructed using pdKCR and pdBPV-1 plasmid and transfected into mouse C127 cells by calcium phosphate coprecipitation. Transfected cells exhibited altered cell morphology, could be continuously cultured in DMEM essential medium and secreted IL-4 lymphokine into 1% FCS condition culture medium. High biological activity (2 x 10(4)/ml) from the culture supernatant was obtained by FACS analysis of Fc epsilon receptor expression on B cell surface.

Animals↗

Phenotypical and functional analyses of natural killer cells from low NK activity individuals among healthy and patient populations.

We investigated the deficiency of natural killer (NK) activity by contrasting healthy individuals with patients. Human NK activities of 125 individuals consisting of 68 healthy donors and 57 patients (36 autoimmune disease and 21 cancer patients) were measured by Eu-DTPA release assay in which the target cells were labeled by nonradioactive materials-Eu-DTPA, and they were phenotypically analyzed with three-color flow cytometry. Furthermore, a part of these donors was functionally studied on NK cells sorted out from PBL. 23.3% of healthy donors and approximately 70% of patients had low NK activity (LNK). In these healthy LNK and patient LNK, the population of CD3-CD16+CD56+ subset in PBL was significantly lower than that of the same subset in healthy individuals with high and medium NK activity (HMNK). The cytotoxicity of CD3-CD16+CD56+ cells sorted out from PBL in healthy LNK and patient LNK were approximately the same with or higher than that in healthy HMNK. No differences were found either in the expression of CD2 and LFA-1 antigens on the CD3-CD56+ NK cells or in the amount of granulous proteins such as perforin and granzyme A in these cells among healthy HMNK, healthy LNK and patient LNK. These results suggested that low NK activity of healthy LNK and patient LNK was more reflected by the diminution of the population of CD3-CD16+CD56+ subset in PBL rather than the functional defects of NK cells. A phenotypical and functional study on healthy LNK has not been reported extensively, and we found several differences between healthy LNK and patient LNK in this study. By stimulation with IL-2, the cytotoxicity of healthy LNK increased more rapidly than that of patient LNK, and at high effector:target cell ratio (> or = 40) it was significantly higher than that of patient LNK. The population of CD3+CD16+/CD56+ subset in PBL of healthy LNK was higher than that of patient LNK, but on the other hand it was about the same as that of healthy HMNK.

Adult↗

Elimination of cocaine-induced hyperactivity and dopamine-mediated neurophysiological effects in dopamine D1 receptor mutant mice.

The brain mesoaccumbens dopamine system is intricately involved in the psychomotor stimulant activities of cocaine. However, the extent to which different dopamine receptors mediate these effects has not yet been firmly established. The present study used dopamine D1 receptor mutant mice produced by gene targeting to investigate the role of this receptor in the effects induced by cocaine. In contrast with wild-type mice, which showed a dose-dependent increase in locomotion, D1 mutant mice exhibited a dose-dependent decrease. Electrophysiological studies of dopamine-sensitive nucleus accumbens neurons demonstrated a marked reduction in the inhibitory effects of cocaine on the generation of action potentials. In addition, the inhibitory effects of dopamine as well as D1 and D2 agonists were almost completely abolished, whereas those of serotonin were unaffected. D2-like dopamine receptor binding was also normal. These results demonstrate the essential role of the D1 receptor in the locomotor stimulant effects of cocaine and in dopamine-mediated neurophysiological effects within the nucleus accumbens.

Animals↗

Dopamine D1 receptor mutant mice are deficient in striatal expression of dynorphin and in dopamine-mediated behavioral responses.

The brain dopaminergic system is a critical modulator of basal ganglia function and plasticity. To investigate the contribution of the dopamine D1 receptor to this modulation, we have used gene targeting technology to generate D1 receptor mutant mice. Histological analyses suggested that there are no major changes in general anatomy of the mutant mouse brains, but indicated that the expression of dynorphin is greatly reduced in the striatum and related regions of the basal ganglia. The mutant mice do not respond to the stimulant and suppressive effects of D1 receptor agonists and antagonists, respectively, and they exhibit locomotor hyperactivity. These results suggest that the D1 receptor regulates the neurochemical architecture of the striatum and is critical for the normal expression of motor activity.

Analysis of Variance↗

Activation of the Ig germ-line gamma 1 promoter. Involvement of C/enhancer-binding protein transcription factors and their possible interaction with an NF-IL-4 site.

Ig isotype switching in B lymphocytes is preceded by transcription of the corresponding unrearranged, or germ-line (GL), CH gene. The promoter of mouse GL C gamma 1 transcripts has been shown to be located within a 349-bp KpnI/Bg/II fragment, extending from -147 to +202 bp relative to the first transcription initiation site. By the electrophoretic mobility shift assay, we have analyzed nuclear extracts from three B cell lines and splenic B cells for the presence of proteins binding to this fragment. We show that they give different patterns of DNA binding, implying significant complexity in the regulation of this locus. We focused on the sIgM+ mouse B lymphoma line L10A6.2 that has been shown able to confer responsiveness of the GL gamma 1 promoter to phorbol ester plus IL-4. Activation of this cell line results in altered expression of several nuclear DNA-binding complexes involving two members of the C/enhancer-binding protein (EBP) family of transcription factors, namely C/EBP beta (nuclear factor (NF)-IL-6/LAP) and Ig/EBP-1 (C/EBP gamma). The complexes bind to two C/EBP elements, one at about -115 bp and one near the first RNA start site. In normal B cells stimulated by LPS or IL-4, new complexes appear that bind to C/EBP and NF-IL-4 elements, respectively, located within the -125/-101 region. The -125/-101 segment previously has been shown to be required for transcriptional activation. We discuss these findings in relation to the presence of consensus C/EBP binding sites in other IL-4-regulated promoters.

Animals↗

More efficient peptide binding to MHC class II molecules during cathepsin B digestion of Ii than after Ii release.

The binding of a T cell-presented peptide to MHC class II alpha,beta chains occurs as a concurrent process with the release of the associated invariant chain (Ii) by cathepsin B. Ii was digested by cathepsin B from solubilized, MHC class II alpha,beta,Ii complexes in the presence of N-hydroxysuccinimidyl-4-azidobenzoate-conjugated, 125I-labeled, influenza virus matrix (18-29) peptide. The peptide was crosslinked where it became bound. This HLA-DR1-restricted peptide bound about three times more efficiently to class II alpha,beta chains of DR1-positive B cells when present during cathepsin B digestion of Ii than when added afterward, also at pH 5.0. Binding was competed by similarly DR-restricted peptides. Cathepsin D cleaved Ii but did not enhance peptide binding. However, a trace level of cathepsin D, added to the assay for peptide binding in the presence of cathepsin B, further enhanced peptide binding about three times. These experiments support an hypothesis for the staged release of Ii fragments by cathepsin D and cathepsin B, catalyzing at one point the insertion of a peptide into the antigen binding site formed by class II alpha and beta chains.

Amino Acid Sequence↗

Cathepsin B cleavage and release of invariant chain from MHC class II molecules follow a staged pattern.

A staged pattern of cathepsin B cleavage of MHC class II alpha, beta-bound invariant (Ii) chain and release of fragments was defined. Charge-loss mutations in the Ii chain were created in three clusters of cathepsin B putative cleavage sites R78K80K83K86, K137K143, and R151K154. Products of HLA-DR1 alpha, beta and wild type (WT) or mutant Ii genes, co-transfected into COS1 cells, were cleaved by cathepsin B and immunoprecipitated by antibodies either to MHC class II chains or to different Ii epitopes. In WT Ii, cathepsin B digestion generated two forms of p21 Ii fragments: a p21 recognized by anti-C-terminus antibodies and a p21 recognized by an antibody to a determinant near the N-terminus. C-terminal p21 was released from MHC class II alpha, beta chains upon its formation while N-terminal p21 remained associated with MHC class II alpha, beta chains. Mutations at K137K143 inhibited the generation of N-terminal p21 by cathepsin B. Mutation at R78K80K83K86 led to an accumulation of MHC class II-bound N-terminal p21 without the appearance of MHC class II-bound p14, p10, and p6 fragments after cathepsin B digestion. These results indicate that cathepsin B cleaves wild type Ii first about K137K143 to produce a MHC class II-associated N-terminal p21, which is then cleaved about R78K80K83K86 to generate p14, p10 and finally p6 which still associates with MHC class II alpha, beta chains. This pattern of staged cleavage and release of Ii might be related to a concerted mechanism regulating the binding of antigenic peptides to MHC class II molecules.

Amino Acid Sequence↗

Demonstration and characterization of simultaneous production of a thermostable direct hemolysin (TDH/I) and a TDH-related hemolysin (TRHx) by a clinically isolated Vibrio parahaemolyticus strain, TH3766.

Simultaneous production of a thermostable direct hemolysin (TDH)-like toxin (TDHx) and a TDH-related hemolysin (TRH)-like toxin (TRHx) by a clinical isolate (strain TH3766) of Kanagawa phenomenon-positive Vibrio parahaemolyticus was demonstrated and characterized. The two hemolysins were differentially purified by column chromatography on hydroxyapatite and immunoaffinity columns. The molecular weight of the two hemolysins were estimated to be 23,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE). The purified TDHx was indistinguishable from the previously reported TDH/I (from strain TH012) but was different from the authentic TDH of a Kanagawa phenomenon-positive strain (T4750) physicochemically. The mobility of TRHx in nondenaturing PAGE differed from all the known TDHs and TRHs. The genes (tdhX and trhX) coding for TDHx and TRHx were cloned and sequenced. Homologies of nucleotide sequences of the coding regions between tdhX and tdhA (a gene for the authentic TDH) and between trhX and trh (a gene for the authentic TRH) were 98.1 and 99.1%, respectively, and homology between tdhX and trhX was 68.1%. At the amino acid level, TdhX was completely identical to TDH/I, although two base differences were found in the nucleotide sequences between tdhX and tdh/I. Two amino acid differences were observed between TrhX and Trh. Thus, these findings suggest that the TH3766 strain produces two types of hemolysins simultaneously. This is the first evidence that a strain of V. parahaemolyticus produces two types of toxins of the TDH-TRH family at the same time.

Amino Acid Sequence↗

Construction and characterization of an isogenic mutant of Vibrio parahaemolyticus having a deletion in the thermostable direct hemolysin-related hemolysin gene (trh)

A mutant, Vp-MX02, having a deletion in the gene (trh) for the thermostable direct hemolysin-related hemolysin (TRH) was constructed by double-crossover gene conversion from a TRH-producing Vibrio parahaemolyticus strain, TH3996. The deleted region was the upstream half of trh. Hemolysis was completely lost, and TRH antigen was undetectable in the mutant. Administration of the mutant into ligated rabbit small intestines elicited partial, but apparent, fluid accumulation. These results suggest that the enterotoxicity of TRH-producing V. parahaemolyticus TH3996 may be attributed to the remaining C-terminal of TRH or, more likely, to an unknown virulence factor(s) in addition to TRH.

Animals↗