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Biomedical subjects

Wei Zhou

Publications and source records attributed to Wei Zhou.

At least 199 records · Page 11Linked to original sources

TLR4 Asp299Gly and Thr399Ile polymorphisms are very rare in the Chinese population.

Endotoxin is an important component of bio-aerosols that contribute to airway inflammation and airflow obstruction. Toll-like receptor 4 (TLR4) mediates the host response to bacterial lipopolysaccharide (LPS), and the Asp299Gly and Thr399Ile polymorphisms have been associated with the development of respiratory diseases. We hypothesized that TLR4 polymorphisms may be associated with a decline of lung function in cotton workers who were exposed continuously to endotoxin. We investigated these two polymorphisms in our 20-year longitudinal cohort of Han Chinese cotton and silk textile workers. The two polymorphisms were genotyped using TaqMan and PCR-restriction fragment length polymorphism methods. No homozygous or heterozygous variant genotypes of the Asp299Gly and Thr339Ile polymorphisms were detected in 491 samples of this population. Our results suggest that the TLR4 Asp299Gly and Thr339Ile polymorphisms are very rare in the Chinese population.

Adult↗

Reevaluation of the conclusion that IRES-activity reported within the 5' leader of the TIF4631 gene is due to promoter activity.

We previously reported that the 5' leader of the mRNA-encoding initiation factor eIF4G in Saccharomyces cerevisiae can function as a translational enhancer and as an internal ribosome entry site (IRES) when tested in cells. However, Verge and colleagues recently suggested that this sequence does not facilitate translation initiation, but inhibits translation in vitro and has promoter activity when tested in cells. We disagree with these conclusions and respond by showing that the data are most consistent with an internal initiation mechanism.

5' Untranslated Regions↗

[Respiratory distress syndrome induced by immersion in seawater: a study of canine models].

OBJECTIVE: To study the mechanism of respiratory distress syndrome (RDS) induced by immersion in seawater to provide experimental evidence for its treatment. METHODS: Twelve normal hybrid dogs were randomly assigned into control group (n=4) and SDS model group (n=8). The changes in blood dynamics, blood gas analysis and histological changes in the lung tissues were compared between the 2 groups. The concentration of lactic dehydrogenase (LDH-L) and alkaline phosphatase (ALP) in the bronchoalveolar fluid and blood of the dogs in the model group were tested. RESULTS: The blood dynamics, blood gas analysis and histology of the dogs in the model group were significantly different from those in the control group, and LDH-L and ALP levels increased significantly in the bronchoalveolar fluid of the model group. CONCLUSION: Seawater aspiration into the lungs may lead to RDS, and the canine models used in this study may help explore the mechanism and management of RDS induced by immersion in seawater.

Alkaline Phosphatase↗

[Polymorphism in intron 2 of crucian carp GH I gene].

High length and nucleotide polymorphisms in intron2 of GH I gene were detected in 162 individuals,which were from seven wild crucian carp colonies, two goldfish colonies and one Fangzheng crucian carp colony. Using denaturating polyareylamide gel electrophoresis (DPAGE) and single-strand conformation polymorphism (SSCP), seven length variants and 15 haplotypes were identified in these fishes. The length types and haplotypes diversity was 4.32% and 9.26%, respectively. Sequence analysis of the 15 haplotypes indicated the following results: (1) The size of intron2 varied from 243 to 263 bp. In the 15 haplotypes,the average percentages of the four bases (A,T,G and C) were 34.13%, 37.36%, 15.13% and 13.38%, respectively; the frequency of G + C(28.51%) was much lower than that of A + T (71.49%). The GT/AG domain was found in exon-intron junctions,which was the 5' and 3' splice donor and acceptor sites in higher eukaryotic gene introns. The similarity sequence of GTAAGTA was located on the junction between exon2 and intron2. And there existed a richer pyrimidine region (TTTGCCTTTTGTTATC) near the 3' end of intron2. (2) The seven length variants (A, B, C, D, E, F and G) were determined to be 189, 196, 204, 205, 206, 207 and 209 bp, respectively. The polymorphism resulted from the variable repeat number of T (N = 0, 8, 9, 10, 11 and 13) and the difference in one or two motifs deletions of TGAAAAC, TT and GAGTG. (3) Compared the sequences of the 15 haplotypes, 17 substitution sites were observed, of which two were of transversion sites and 15 were of transition sites. Obviously, the transition mutations (88.24%) were more frequent than transversion mutations (11.76%). Analysis of the distributions of the length types, haplotypes and composite genotypes suggested that genetic diversity was varied in different colonies. In the goldfish colonies, only one length type (A), two haplotypes (A1 and A2) and one composite genotype (A1A2) were observed. Two length types (C and D), four haplotypes (C1, C2, D2 and D5) and one composite genotype (C1C2D2D5) presented in the Fangzheng crucian carp colonies. The highest level of genetic diversity was exhibited in the seven wild crucian carp colonies: seven length types (A, B, C, D, E, F and G), 14 haplotypes (A1, A2, A3, B, C1, C2, D1, D2, D3, D4, E1, E2, F and G) and 14 composite genotypes (A1A2A3, A1A2A3B, A1C1C2D1D2D3, A1C1C2D2, A1C1C2D2D3, A1C1C2D3E2, BC1C2D2, BC1C2D3D4, C1C2D2, C1C2D2D3, C1C2D3D4, C1C2D3D4F, C1C2D4, D2E1G) were shared by the seven wild colonies. The numbers of observed length types, haplotypes and genotypes within the wild colonies ranged from 3 to 6, 6 to 10 and 2 to 6, respectively. Whether the length and nucleotide polymorphisms in the intron2 of crucian carp GH I gene were associated with gene expression and gene regulation remained unsolved and required further investigations.

Animals↗

[Management of hyperkalemia-induced cardiac arrest in anhepatic stage of orthotopic liver transplantation without venovenous bypass: report of 3 cases].

The causes of high serum potassium-induced cardiac arrest in anhepatic stage of orthotopic liver transplantation were analyzed in 3 cases without venovenous bypass, and the effectiveness of insulin was evaluated in correcting hyperkalemia during the anhepatic stage. To improve the survival rate of patients with such cardiac arrest, early cardiopulmonary resuscitation and timely defibrillation should be performed.

Adult↗

[The role of circadian gene period1 in morphine reward in mice].

OBJECTIVE: To investigate the role of circadian gene Period1 on drug dependence. METHOD: The ribozyme cleave the Period1 mRNA (per1RZ) was designed and the per1RZ expression vector (pcDNA 3.1-per1RZ) was constructed based on eukaryotic expression vector pcDNA 3.1. In vitro cleavage experiment proved the target cleave Period1 mRNA ability of per1RZ. Conditional place preference (CPP) paradigm used to investigate the effect of intracerebroventricular (ICV) injection of pcDNA 3.1-per1RZ on drug reward in BALB/C mice. RESULT: Quantitative analysis of in vitro cleavage experiment showed the efficacy of ribozyme per1RZ, about 60% of the Period1 mRNA was cleaved by ribozyme per1RZ. CPP test displayed the block of drug reward in pcDNA 3.1-per1RZ ICV injection group. Period1 expression in brain was attenuated of pcDNA 3.1-per1RZ ICV injection group demonstrated by western blot. CONCLUSION: Interfere the Period1 expression in brain could attenuate the psychological dependence of drug in mammals.

Animals↗

Effective elicitation of anti-tumor immunity by collocation of antigen with encoding gene in the same vaccine.

Peptide and naked DNA vaccines, aimed at generating strong cytotoxic T lymphocyte (CTL) responses capable of mediating tumor regression, have been considered of the most rapidly evolving technologies for tumor vaccination. Results from clinical trials of these strategies are encouraging, but unfortunately, most of those trials have been proved as partly successful. Given the distinct dynamics of antigen presentation for peptide and gene forms of antigens, we explored a novel concept that collocation of the antigen with the encoding gene in the same vaccine could effectively elicit anti-tumor immunity, and developed a novel peptide-DNA dual vaccine (PDDV), which combines the benefits of peptide and DNA vaccines and could induce tumor-specific CTL response. Furthermore, PDDV effectively protected mice against fatal P815 tumor challenge and cured tumor-bearing DBA/2 mice, suggesting PDDV as a potential formulation of tumor vaccine.

Animals↗

Oral immunization with rotavirus VP7 expressed in transgenic potatoes induced high titers of mucosal neutralizing IgA.

Rotaviruses (RV) are a common cause of severe diarrhea in young children, resulting in nearly one million deaths worldwide annually. Rotavirus VP7 was the rotavirus neutralizing protein. Previous study reported that VP7 DNA vaccine can induce high levels of IgG in mice but cannot protect mice against challenge (Choi, A.H., Basu, M., Rae, M.N., McNeal, M.M., Ward, R.L., 1998. Virology 250, 230-240). We found that rotavirus VP7 could maintain its neutralizing immunity when it was transformed into the potato genome. Mice immunized with the transformed tubers successfully elicited serum IgG and mucosal IgA specific for VP7. The mucosal IgA titer was as high as 1000, while serum IgG titer was only 600. Neutralizing assays indicated that IgA could neutralize rotavirus. These results indicate the potential usefulness of plants for production and delivery of edible rotavirus vaccines.

Administration, Oral↗

Nanoporous carbide-derived carbon with tunable pore size.

Porous solids are of great technological importance due to their ability to interact with gases and liquids not only at the surface, but throughout their bulk. Although large pores can be produced and well controlled in a variety of materials, nanopores in the range of 2 nm and below (micropores, according to IUPAC classification) are usually achieved only in carbons or zeolites. To date, major efforts in the field of porous materials have been directed towards control of the size, shape and uniformity of the pores. Here we demonstrate that porosity of carbide-derived carbons (CDCs) can be tuned with subångström accuracy in a wide range by controlling the chlorination temperature. CDC produced from Ti3SiC2 has a narrower pore-size distribution than single-wall carbon nanotubes or activated carbons; its pore-size distribution is comparable to that of zeolites. CDCs are produced at temperatures from 200-1,200 degrees C as a powder, a coating, a membrane or parts with near-final shapes, with or without mesopores. They can find applications in molecular sieves, gas storage, catalysts, adsorbents, battery electrodes, supercapacitors, water/air filters and medical devices.

Carbon↗

Dual regulation of proliferation and growth arrest in prostatic stromal cells by transforming growth factor-beta1.

In a preliminary study, we observed that TGF-beta1 induced both proliferation and growth arrest in prostatic stromal cells, depending on the concentration of TGF-beta1 used in the culture medium. In this study, we explored possible mechanisms of this dual effect of TGF-beta. Primary cultures of prostatic stromal cells, established from clinical surgical specimens and treated with low doses of TGF-beta1 (0.001-0.01 ng/ml), resulted in an increase in cell proliferation. The addition of neutralizing antibody against platelet-derived growth factor (PDGF)-BB, but not anti-PDGF-AA, abrogated this stimulatory effect of TGF-beta1. TGF-beta1 treatment resulted in a dose-related increase in PDGF-BB production as measured by ELISA. Cells underwent growth arrest at high concentrations of TGF-beta1 (1.0 and 10 ng/ml). An inhibitor of cyclin-dependent kinase (cdk), p15INK4b, was up-regulated at both transcript and protein levels in these cultures by TGF-beta1 in a dose-related manner as determined by RT-PCR and Western blot analysis. The transcript, but not the protein, for another cdk inhibitor, p21Cip1, was up-regulated with treatment of TGF-beta1 to these cells. Levels of other cdk inhibitors, such as p16INK4a and p27Kip1, were constitutively expressed in prostatic stromal cells and were not significantly affected by TGF-beta1 treatment. Finally, the growth arrest effect of TGF-beta1 was abrogated when antisense oligonucleotides to p15INH4b, but not p21Cip1, were added to the culture medium. These data indicate that the dual effect of TGF-beta1 is mediated, at least, by up-regulation of PDGF-BB and p15INK4b, respectively.

Antibodies↗

Determining the basis of channel-tetramerization specificity by x-ray crystallography and a sequence-comparison algorithm: Family Values (FamVal).

We have developed a semiempirical algorithm called Family Values (FamVal), which identifies residues that encode functional specificity in a protein sequence. Given a multiple sequence alignment (MSA) grouped into functionally distinct subfamilies, FamVal calculates a specificity score for each subfamily at every amino acid position of an MSA. This algorithm was used to predict specificity-encoding positions within the tetramerization assembly (T1) domain of voltage-gated potassium (Kv) channel subfamilies Kv3 and Kv4. The importance of one such position (Arg to Ala at MSA position 93) was confirmed by in vitro pull-down assays. The structural basis of this assembly discrimination was elucidated by determining the crystal structure of the Kv4 T1 domain and comparing it to the Kv3 T1 domain.

Algorithms↗

Isolation and identification of short nucleotide sequences that affect translation initiation in Saccharomyces cerevisiae.

In previous studies, we demonstrated the sufficiency of short nucleotide sequences to facilitate internal initiation of translation in mammalian cells. By using a selection methodology, we have now identified comparable sequences in Saccharomyces cerevisiae. For these studies, a library of constructs expressing dicistronic mRNAs with the HIS3 gene as the second cistron and 18 random nucleotides in the intercistronic region was introduced into a yeast strain in which the endogenous HIS3 gene was deleted. Untransformed cells or those containing the parent construct failed to grow on medium lacking histidine. Intercistronic sequences recovered from cells that did grow were evaluated by using various criteria. Fifty-six of the 18-nt sequences (approximately 1/400,000) functioned as synthetic internal ribosome entry sites (IRESes). The 14 most active sequences allowed growth in the presence of 0.1-0.6 mM 3-amino-1,2,4-triazole, a competitive inhibitor of the HIS3 gene product. In addition, eight sequences were identified that were not IRESes, but that enhanced HIS3 expression by an alternative mechanism that depended on the 5' end of the mRNA and appeared to involve either shunting or reinitiation. Comparisons among the 56 selected IRESes identified eight significant sequence matches containing up to 10 nucleotides. Many of the selected sequences also contained extensive complementary matches to yeast 18S rRNA, some at overlapping sites. The identification of cis sequences that facilitate translation initiation in yeast enables detailed biochemical and genetic analyses of underlying mechanisms and may have practical applications for bioengineering.

Base Sequence↗

P53 (codon 72) and P21 (codon 31) polymorphisms alter in vivo mRNA expression of p21.

p21 (Waf1/Cip1) is a downstream target of p53. We evaluated the association between p21 polymorphism (codon 31), p53 polymorphism (codon 72) and their corresponding in vivo mRNA expression. In this study, p21 and p53 genetic polymorphisms (using standard PCR-RFLP techniques) and p21 and p53 gene expressions (using a radiolabelled ribonuclease protection assay (RPA) technique) were evaluated in the peripheral leukocytes of 84 individuals (63 with lung cancer). Log-transformed values of mRNA expression by RPA, which approximated a normal distribution, were analyzed. p53 genotypes did not correlate with p53 mRNA log-expression (P>0.05 for all comparisons), but the Pro allele variants of p53 were associated with a significant decrease in mRNA log-expression of its downstream target, p21. The variant Arg allele of p21 was also associated with a significant decrease in p21 mRNA log-expression. When individuals with at least one variant allele of both p53 and p21 (double-variants) were compared with all other genotype groups, these double-variants had significantly lower log-expression of p21 (P<0.005 by both t-tests (crude) and linear regression analyses (adjusted)). This is translated into an approximate 48% reduction in the geometric mean of the mRNA expression of the double-variants, when compared with all other groups. Results were consistent in both patients with lung cancer (n=63) and in normal controls (n=21). In conclusion, the presence of a p53 Pro allele and/or p21 Arg allele is associated with lower downstream target gene expression of p21.

Aged↗

Mapping genetic alterations in tumors with single nucleotide polymorphisms.

Single nucleotide polymorphisms are the most abundant genetic markers in the human genome whose analyses can be easily conducted on a large scale. Most established methods for high-throughput single nucleotide polymorphism analyses are qualitative and are not suitable for genetic analysis of archived tumor specimens, which have compromised tissue integrity and normal tissue contaminations. Recent studies have focused on the development of quantitative methods for single nucleotide polymorphism analyses that can tolerate such imperfections in archived tissues. These methods have been used to measure the rate of allelic imbalance in small adenocarcinomas and to reveal novel correlations between allelic imbalance and disease progression in colorectal cancer. Therefore, quantitative single nucleotide polymorphism analysis provides a powerful tool for the identification of novel tumor markers and for the characterization of genetic alterations in human tumors.

Allelic Imbalance↗

Genotyping patients with recent blood transfusions.

BACKGROUND: Many studies have used polymerase chain reaction amplification (PCR) to genotype for common polymorphisms in intensive-care units (ICUs) where blood transfusions are common. Evidence that donor leukocytes in transfused blood can be detected by PCR of the recipient blood suggests that this minor population of donor white cells (microchimerism) can interfere with genotyping of allelic polymorphisms in critically ill transfused patients. To investigate this possibility, we assayed DNA extracted from the blood and buccal cells of ICU patients for 2 common polymorphisms in the TNF-beta gene and the surfactant protein-B (SP-B) gene. METHODS: Study subjects were ICU patients from the Massachusetts General Hospital (Boston, MA) enrolled into a study on the molecular epidemiology of acute respiratory distress syndrome between January 1999 and October 2000. Blood and buccal cells were collected and DNA was extracted from 145 patients. Genotyping was performed by enzyme digestion and pyrosequencing. RESULTS: The Kappa statistics comparing the genotype results from blood and buccal cells were 0.98 (95% confidence interval [CI] = 0.94-1.01) for TNFB and 0.95 (CI = 0.87-1.02) for SP-B. When the analysis was restricted only to the 107 patients who were transfused, the Kappa statistic remained high at 0.97 (CI = 0.93-1.01) for TNFB and 0.93 (CI = 0.84-1.03) for SP-B. CONCLUSION: We conclude that microchimerism from allogeneic blood transfusion is unlikely to have major effects on the genotype results of common polymorphisms in large molecular epidemiology studies conducted in the critical care setting if DNA is collected within a day after transfusions.

Alleles↗