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Biomedical subjects

X Zhou

Publications and source records attributed to X Zhou.

At least 253 records · Page 14Linked to original sources

Efficient translocation and processing with Xenopus egg extracts of proteins synthesized in rabbit reticulocyte lysate.

Cell-free translation/translocation systems are broadly applied to examine gene expression and characterize the structure-function relationship of gene products. We present the characterization of Xenopus egg extract (XEE) translocation and processing of proteins synthesized in rabbit reticulocyte lysate. The XEE was prepared from eggs laid by adult female frogs that received serial injections of gonadotropins. The eggs were then dejellied in 2% L-cysteine-HCl and the cytoplasm extracted by centrifugation at 10,000 rpm for 15 min. The in vitro translocation and processing of XEE was examined with a cell-free translation system containing reticulocyte lysate, and appropriate messenger ribonucleic acid (RNA) or complementary deoxyribonucleic acid plasmids with RNA polymerase. Cell-free production of the following proteins were used to assess posttranslational modifications: Escherichia coli beta-lactamase for signal sequence cleavage, Saccharomyces cerevisiae alpha-mating factor for translocation and N-linked glycosylation, the soluble protein luciferase for functional activity, and the membrane-bound human insulin receptor for translation efficiency. All translation products were identified by [35S]-methionine labeling, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. The results demonstrate that (1) XEE produces near-complete signal sequence and N-glycosylation processing of proteins synthesized in reticulocyte lysate, (2) XEE contains endoplasmic reticulum-equivalent microsomes, which allows for protein translocation and protease protection, (3) the addition of XEE in the translation reaction does not affect synthesis and chemiluminescence activity of luciferase, (4) XEE is efficient in processing the nascent 160-kDa human insulin receptor precursor, a transmembrane protein, and (5) as compared to canine pancreatic microsomes, XEE translocation efficiency is minimally decreased with the addition of dimethylsulfoxide. These results are the first description of the combined use of XEE with reticulocyte lysate and clearly demonstrate a higher efficiency of translocation and processing compared to canine pancreatic microsomes. This method of cell-free translation and processing allows for more extensive in vitro examination of posttranslational modifications of secretory and membrane-bound proteins.

Animals↗

Biological activity of CD-ring modified 1alpha,25-dihydroxyvitamin D analogues: C-ring and five-membered D-ring analogues.

Nonsteroidal analogues of 1alpha,25(OH)2D3, lacking either the full five-membered D ring (C-ring analogues) or the full six-membered C ring (D-ring analogues) are more potent inhibitors of cell proliferation or inducers of cell differentiation than is 1alpha,25(OH)2D3. Maximal superagonistic activity was seen for the C-ring analogue with a 24(R)-hydroxyl group in the side chain [30- to 60-fold the activity of 1alpha,25(OH)2D3]. The 19-nor-16-ene-26,27-bishomo C-ring analogue showed the best ratio of antiproliferative to calcemic effects (1275-fold better than 1alpha,25(OH)2D3 and severalfold better than all vitamin D analogues so far described). The analogues are able to stimulate specific vitamin D-dependent genes and are active in transfection assays using an osteocalcin promoter VDRE. Low binding affinity to the vitamin D binding protein, differences in metabolism, or affinity for the vitamin D receptor (VDR) are not the most important explanations for the enhanced intrinsic activity. However, the analogues are able to induce conformational changes in the VDR, which makes the VDR-ligand complex more resistant against protease digestion than is 1alpha,25(OH)2D3. In contrast to 20-epimer steroidal vitamin D analogues, 20-epimer C-ring analogues were less potent than analogues with a natural C-20 configuration. In conclusion, several nonsteroidal vitamin D analogues are superagonists of 1alpha,25(OH)2D3 despite lower receptor affinity and, for the C-ring analogues, higher flexibility of the side chain; moreover, they have a better selectivity profile than all analogues yet published.

Calcitriol↗

Larval susceptibility of an insecticide-resistant western corn rootworm (Coleoptera: Chrysomelidae) population to soil insecticides: laboratory bioassays, assays of detoxification enzymes, and field performance.

Soil insecticides were evaluated in laboratory and field studies against larvae of an insecticide resistant population (Phelps County, NE) of western corn rootworm, Diabrotica virgifera virgifera LeConte. Insecticide toxicity was evaluated by topical application of technical insecticides to 3rd instars from Saunders County, NE (susceptible) and Phelps County populations. Resistance ratios (LD50 Phelps County/LD50 Saunders County) for the insecticides methyl parathion, tefluthrin, carbofuran, terbufos, and chlorpyrifos were 28.0, 9.3, 8.7, 2.6 and 1.3, respectively. Biochemical investigation of suspected enzymatic resistance mechanisms in 3rd instars identified significant elevation of esterase activity (alpha and beta naphthyl acetate hydrolysis [3.8- and 3.9-fold]). Examination of 3rd instar esterases by native PAGE identified increased intensity of several isoenzymes in the resistant population. Assays of cytochrome P450 activity (4-CNMA demethylation and aldrin epoxidation) did not identify elevated activity in resistant 3rd instars. Granular soil insecticides were applied at planting to corn, Zea mays L., in replicated field trials in 1997 and 1998 at the same Phelps County site as the source of resistant rootworms for the laboratory studies. In 1997, planting time applications of Counter 20CR, Counter 15 G (terbufos), and Lorsban 15 G (chlorpyrifos) resulted in the lowest root injury ratings (1-6 Iowa scale); 2.50, 2.55, 2.65, respectively (untreated check root rating of 4.55). In 1998, all insecticides performed similarly against a lower rootworm density (untreated check root rating of 3.72). These studies suggest that resistance previously documented in adults also is present in 3rd instars, esterases are possibly involved as resistance mechanisms, and resistance to methyl parathion in adults is also evident in larvae, but does not confer cross-resistance in larvae to all organophosphate insecticides.

Animals↗

Antimicrobial activity of cetylpyridinium chloride washes against pathogenic bacteria on beef surfaces.

Cetylpyridinium chloride (CPC), a water-soluble, neutral pH, colorless compound, is widely used in oral hygiene products to inhibit bacteria responsible for plaque. Previously, researchers have demonstrated that CPC not only reduces Salmonella typhimurium on poultry but also prevents cross-contamination. To determine the effectiveness of CPC against pathogens associated with lean and adipose beef surfaces, several spray-washing experiments (862 kPa, 15 s, 35 degrees C) with 1% (wt/vol) CPC were conducted. On lean beef surfaces, CPC immediately reduced 5 to 6 log10 CFU/cm2 of Escherichia coli O157:H7 and Salmonella typhimurium to virtually undetectable levels (0 log10 CFU/cm2), as well as after 35 days of refrigerated (4 degrees C), vacuum-packaged storage. On adipose beef surfaces, 5 log10 CFU/cm2 Salmonella typhimurium and E. coli O157:H7 were reduced immediately (>2.5 log10 CFU/cm2) with 1% CPC; by day 35 the reduction was <1.3 log10 CFU/cm2. Further plate overlay analyses indicated that the effectiveness of CPC against pathogens on adipose surfaces was not hampered by the presence of meat components or fatty acids. Additional chemical and microbiological analyses of 1% CPC-treated beef surfaces subjected to a secondary water wash (following contact times of 0, 5, 10, 15, or 30 min) or grinding did reduce pathogenic bacteria and CPC levels. However, residual CPC levels following any of the treatments were considered excessive for human consumption. Despite the residual levels, this study is the first to demonstrate the effect of CPC on pathogenic bacteria associated with beef surfaces immediately after treatment and also after long-term, refrigerated, vacuum-packaged storage.

Animals↗

Prognostic implication of microsatellite alteration profiles in early-stage non-small cell lung cancer.

Development of non-small cell lung cancer (NSCLC) is a result of multiple accumulated genetic abnormalities. Profiles of genetic abnormalities may determine tumor behavior and impact on patient outcome. We used microsatellite markers at 3p14, 9p21, and 10q24 to analyze tumor samples from 91 patients with pathologically confirmed stage I NSCLC for microsatellite alterations. Loss of heterozygosity at any single locus was not significantly associated with length of survival. However, patients whose tumors had microsatellite instability (MI) at 10q24 had shortened disease-specific survival. Among 31 such patients, 32% (10 of 31 patients) had died of the disease within 5 years after surgery compared with 16% (9 of 58 patients) without MI at 10q24 (P = 0.07). Interestingly, in the adenocarcinoma subtype, 71% (5 of 7 patients) of the patients with MI at 10q24 succumbed to the disease as compared with only 12% (3 of 26) of the adenocarcinoma patients without such MI (P < 0.001), suggesting the presence of distinct mechanisms in tumorigenesis among different subtypes of lung cancer. It has been noticed that certain microsatellite alteration profiles provide additional values for risk assessment. Of 23 patients who had MI at 10q24 and an alteration at 3p14, 39% (9 of 23 patients) died of the disease within 5 years as compared with only 15% (10 of 66 patients) of the patients without such a profile (P = 0.02). Strikingly, among the 22 patients with no alteration at any loci tested or with loss of heterozygosity at 10q24 and retention of at least one of the other two loci, none died of lung cancer within 5 years after surgery, whereas 28% (19 of 67 patients) of the patients outside these profiles did so (P = 0.01). Our results support the hypothesis that microsatellite alterations can be used as biomarkers for the genetic classification of pathological stage I NSCLC, which may in turn influence treatment decisions dependent on an accurate forecast of patient survival time.

Adenocarcinoma↗

[Progress of study on selectin].

Selectin is a member of vascular adhesion molecular family. Selectin can mediate the interaction between endothelial cell and leucocyte, therefore, it is an important factor in infectious disease, transplantation immunity, development of tumor, vascularization, ischemia-reperfusion injury and autoimmune disease. In this report, selectin's molecular structure, function, expression, regulation and gene reformation were discussed.

Animals↗

[TBX5 mutation in Chinese patients with Holt-Oram syndrome].

OBJECTIVE: To analyse TBX5 mutation in Chinese patients with Holt-Oram syndrome(HOS). METHODS: Seven HOS families were analysed with single strand conformation polymorphism(SSCP) and sequencing. RESULTS: Three SSCP changes were detected and identified as the TBX5 gene mutation at three new sites. One of the changes is a frameshift mutation caused by a base cytidine deletion at the cDNA sequence of 416, which altered all the codons after the point, thus it can not encode the protein of normal amino acid sequence; another is a missense mutation induced by a base substitution(C-->A) at the cDNA sequence of 145, which made the codon of that point change from CAG-->AAG, and encoded amino acid changed from glutamine(Gln) to lysine(Lys), consequently the change weakened the function of TBX5 protein; the third is also a missense mutation which resulted from a base substitution (T-->C) at the cDNA sequence of 161, this change made the codon of that point change from ATC-->ACC, it changed the encoded amino acid from isoleucine(Ile) to threonine(Thr), which reduced the function of TBX5 protein. CONCLUSION: HOS in Chinese is caused by mutation in TBX5.

Abnormalities, Multiple↗

Induction of neonatal tolerance to oxidized lipoprotein reduces atherosclerosis in ApoE knockout mice.

BACKGROUND: In the course of atherosclerosis, humans and apolipoprotein (apoE) Knockout (KO) mice exhibit an active cell-mediated and humoral immune process, both at the systemic level and within atheromata. Low density lipoproteins (LDL) infiltrate the vascular wall, where they are oxidatively modified. This oxidative modification may generate new epitopes for which tolerance is not achieved during ontogenesis. Such epitopes could constitute new targets for autoreactive immune responses that may have a physiopathological role in disease development. MATERIALS AND METHODS: Exposing mice to high dose of antigens during thymic T-cell education induces immunological tolerance to the administered antigens. We injected newborn apoE KO mice with oxidized LDL. They were fed a cholesterol-rich diet and aortic atherosclerosis, cell-mediated immune response, and T-cell repertoire were analyzed after 5 months. RESULTS: Injection of oxidized LDL at birth reduced not only the immune response to oxidized LDL, but also susceptibility to atherosclerosis in apoE mice. Injection of oxidized LDL induced T-cell tolerance due to clonal deletion, rather than anergy of the reactive T cells. The T-cell repertoire of apoE KO mice was affected by the development of the disease, whereas tolerization normalized it. CONCLUSIONS: This study demonstrates that the immune response against oxidized LDL has a deleterious role in atherogenesis and that a fine-tuning of this response could modify the course of the disease.

Animals↗

Intermediate and late rheumatoid arthritis treated by tonifying the kidney, resolving phlegm and removing blood stasis.

Eighty-seven cases of intermediate and late rheumatoid arthritis were treated with Instant Shu Guan Wen Jing Granules ([symbol: see text] Relaxing Joints by Warming Channels) and Instant Shu Guan Qing Luo Granules ([symbol: see text] Relaxing Joints by Removing Heat from the Lung Channel) to tonify the kidney, resolve phlegm and remove blood stasis, and compared with 41 cases treated with Instant Wang Bi Granules ([symbol: see text] Prescription for Arthralgia-syndrome). The treatment produced a clinical cure rate of 54.0% and a total effective rate of 90.8% as in against 29.3% and 73.2% respectively in the control group. The difference was significant (P < 0.01). Improvement in main symptoms and laboratory findings in the treatment group was all more marked than that in the control group (P < 0.05 or P < 0.01), with no side effects observed.

Adult↗

[Preparation and identification of hammerhead ribozyme in vitro against rat caspase-3 mRNA fragment].

OBJECTIVE: To study the transcript effect and cleavage activity in vitro of rat Caspase-3 specific ribozyme (Rz107 and Rz544). METHODS: Rat caspase-3 gene fragment was cloned into T-vector under the control of T7 promoter. (32)P-labeled caspase-3 transcript was target-RNA. Rz107 and Rz544 genes against caspase-3 mRNA were cloned and transcribed in vitro. Cleavage reaction was detected. RESULTS: It was found that Rz107 was active at 37 degrees C and more so at higher temperature within allowing temperature range. The optimal temperature was 50 degrees C. For Rz107, Km and kcat was 14.13 nmol/L and 2.31 min(-1), respectively. However, the Rz544 had no cleavage activity at all. CONCLUSION: Rz107 prepared in vitro possesses the perfect specific catalytic cleavage activity. It is hopeful that Rz107 would be developed to be a new nucleic acid drug that could effectively inhibit the inflammation of hepatitis through cleaving the key gene, caspase-3, in apoptosis in vivo.

Animals↗

[Genetic polymorphism analysis of apoE intron 1 enhancer in patients with coronary heart disease and the association with serum lipid level].

OBJECTIVE: To investigate the association between the polymorphism of apolipoprotein E (apoE) intron 1 enhancer (IE1) and the level of serum lipid in patients with coronary heart disease (CHD). METHODS: ApoE IE1 genotypes of 81 patients with CHD and 108 healthy subjects were detected by polymerase chain reaction and restriction fragment length polymorphism. RESULTS: The frequency of G/G genotype in patients with CHD was significantly higher than that in control group (P < 0.05). The frequency of G allele was significantly higher in patients than in control group (P < 0.05). The gene polymorphism of apoE IE1 obviously influenced the serum lipid levels. The concentrations of TC, and LDL-C among different genotype groups were significantly different, and the level in G/G group was higher than that in C/C group (P < 0.05). CONCLUSION: ApoE IE1 gene polymorphism is closely related to serum lipid, and G/G genotype of apoE IE1 may be a risk factor of coronary heart disease in Han Chinese.

Aged↗

[The role of bcl-2 gene family in taxol-mediated apoptosis in BJAB B cell lymphoma cell line].

OBJECTIVE: To study the apoptosis-inducing effect of taxol on BJAB B cell lymphoma cell line and the role of bcl-2 gene family. METHODS: BJAB cells were treated with taxol and examined for morphologic changes under light and electron microscopes. Flow cytometry assays and DNA gel electrophoresis were used to demonstrate changes in nuclear DNA contents and DNA fragmentations. Changes in bcl-2 gene family at mRNA and protein levels were studied by immunohistochemistry and semi-quantitative RT-PCR technique. RESULTS: Taxol inhibited growth of BJAB cells. The cells initially showed G2/M arrest, followed by apoptosis, in a time and dose dependent manner. The expression of bcl-2 at mRNA and protion level was decreased while that of bcl-xs transcription was increased after taxol treatment. CONCLUSION: Taxol induces apoptosis of BJAB cells. Bcl-2 and bcl-xs are involved in the regulation of taxol-induced apoptosis.

Antineoplastic Agents, Phytogenic↗

Trauma patients with positive cultures have higher levels of circulating macrophage migration inhibitory factor (MIF).

Macrophage migration inhibitory factor (MIF) is a pituitary "stress" hormone that plays a critical role in the host immune response. The aims of the study were to determine whether MIF was detectable in the circulation of trauma patients, to assess whether MIF levels were associated with injury severity, days post injury, infection, and to examine concentrations of other pro-inflammatory cytokines in circulation. We collected plasma samples from 35 trauma (multiple injury) patients and 18 healthy controls. Concentrations of MIF, TNF-alpha, IL-1beta, and IL-6 were measured by ELISA. Average MIF concentration in plasma of trauma patients was 14 fold higher than that of healthy controls (19,439+/-2,615 pg/ml in trauma vs 1,337+/-286 pg/ml in control; p=0.0002). There was no correlation between MIF values and injury severity score or days post injury. Average level of IL-6 in trauma patients was 587+/-85 pg/ml but was not correlated with MIF concentration. TNF-alpha and IL-1beta were not detectable in trauma patients or healthy controls. Higher MIF levels were associated with positive cultures (blood, urine, sputum, wound). These data suggest that MIF may be a possible indicator of infection in trauma patients.

Bacterial Infections↗

Effect of denture adhesive on retention of the mandibular and maxillary dentures during function.

The purpose of this study was to determine the effect of denture adhesive on retention of mandibular and maxillary dentures over a four-hour period. Denture movements were measured using an alternating magnetic field tracking device that determines the position of magnetic receiver coils relative to a transmitter coil positioned over the head. Ten adults with complete maxillary dentures and complete mandibular implant overdentures were enrolled in the study. Specially fabricated mandibular dentures contained a relief area that exposed the implant post which no longer anchored the denture, but now served as an attachment point for a receiver coil that measured mandibular movement. The denture coil was attached lateral to the post coil on a shelf cut into the denture. Mandibular denture movements were recorded as the difference between the mandible movement signal and the mandible + mandibular denture signals. Measurements of denture movements were made at baseline (no adhesive) and at 0, 2, 4 hours post-adhesive application with Fixodent cream for standardized chewing and biting. The MoveTrack signals were recorded on a digital data cassette recorder for subsequent computer analysis. The results of the measurements were analyzed using paired sample t-tests. Specifically, the following comparisons of movement means were made: mandibular vs. maxillary, baseline vs. post-baseline and successive changes (e.g., baseline vs. hour 0, hour 0 vs. hour 2, etc.). Results of these analyses showed that: 1) mandibular denture movements under both adhesive and non-adhesive conditions were significantly greater than maxillary denture movements; 2) the adhesive significantly reduced movement of the maxillary and mandibular dentures during both chewing and biting; and 3) the improvement occurred immediately post-application of the adhesive and was maintained for the four hours of follow-up.

Adhesives↗

[Dynamics of community structure in successional process of needle and broad-leaved mixed forest in Heishiding of Guangdong].

This paper dealt with the dynamics of community structure in a ten-year succession of needle and broad-leaved mixed forest in Heishiding of Guangdong Province. The vertical community structure tended to be complex, while the horizontal one increased by a wide range. There was no big change in species composition, but the dominance of conifers Pinus massoniana and Cunninghamia lanceolata was substituted by the broad-leaved species, such as Schima superba, Symplocos adenophylla and Schefflera octophylla. The whole community changed from needle and boad-leaved mixed forest into broad-leaved and needle mixed forest. The species diversity and evenness of tree layer were increased, the ecological dominance was decreased, and the community became more complicated.

China↗