Antagomir dependent microRNA-205 reduction enhances adhesion ability of human corneal epithelial keratinocytes / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes (NHCEKs).</p><p><b>METHODS</b>Antagomir-205, complementary and inhibitory to microRNA-205, was used to suppress endogenous microRNA-205 in NHCEKs. The adhesion ability of treated NHCEKs was then assessed by cell adhesion assay. Immunoblot and immunohistochemistry were conducted to determine the level of two focal adhesion-related proteins, focal adhesion kinase (FAK) and paxillin (Pax). Phalloidin staining was performed to measure the level of filamentous actin in antagomir-treated NHCEKs.</p><p><b>RESULTS</b>Antagomir-205 markedly reduced the level of microRNA-205 in NHCEKs and significantly enhanced adhesion ability of NHCEKs (P<0.01). Further protein analysis validated that inhibition of microRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax, and decreased filamentous actin.</p><p><b>CONCLUSION</b>Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.</p>

Roles of microRNAs in hematopoietic cell differentiation and the related tumors / 中国医学科学院学报

MicroRNAs (miRNAs) are a family of 21-25 nucleotide small nonprotein-coding RNAs. They regulate gene expression at post-transcriptional level by mRNA degradation or translation repression. Hematopoiesis is one of the most important highly regulated multistage process, which includes orderly turn-on and turn-off of many genes; any wrong modulation may result in blood diseases. Several miRNAs have been found to be involved in hematopoiesis and hematopoietic tumor genesis.

Expression of human HZF1 in E. coli and preparation of antibody against human HZF1 protein / 中国医学科学院学报

<p><b>OBJECTIVE</b>To express human HZF1 fusion protein in E. coli and to obtain an anti-HZF1 antibody.</p><p><b>METHODS</b>A DNA fragment encoding non-zinc finger region of HZF1 protein was inserted into pET30a vector to get the recombination expression plasmid pET30a-HZF1. E. coli was transformed with pET30a-HZF1 and the selected clones were cultured with isopropy-beta-D-thiogalactoside induction. The proteins were prepared from the culture and the fusion protein was purified by Ni column. Rabbits were immunized and reinforced three times with the purified fusion protein. The antiserum was collected and the titer and the specificity of the antibody were checked by ELISA and Western blot.</p><p><b>RESULTS</b>Antibody against HZF1 was obtained and its titer was more than 1:100 000, as proven by ELISA. Western blot analysis showed specific reaction between this antibody and HZF1 fusion protein or the endogenetic HZF1 protein in hemin-induced K562 cells.</p><p><b>CONCLUSIONS</b>The specific antibody against HZF1 is obtained. The antibody may have potential application in farther HZF1 function study and HZF1 determination in tissues and cells.</p>

Gln192Arg polymorphism in paraoxonase 1 gene is associated with Alzheimer disease in a Chinese Han ethnic population / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Oxidative stress such as low-density lipoprotein (LDL) oxidation is thought to be an important mechanism in Alzheimer's disease (AD). Paraoxonase 1 (PON1), an enzyme located on high-density lipoprotein, can prevent LDL from oxidation to some extent. It is also a potent cholinesterase inhibitor and an arylesterase, combating organophosphate poisoning and metabolization of environmental neurotoxins which might be responsible for neurodegeneration with aging. We evaluated the association of Gln192Arg polymorphism in the PON1 gene with AD in a Chinese Han ethnic population.</p><p><b>METHODS</b>Patients and age-matched controls were recruited from outpatient clinics and a population-based epidemiological survey, respectively. Gln192Arg polymorphism in the PON1 gene was detected by allele-specific PCR technique in 521 patients with AD and 578 healthy controls.</p><p><b>RESULTS</b>The presence of at least one of PON1 R alleles (Q/R or R/R) was lower in AD patients than in the controls (82.7% vs 87.4%; chi(2) = 4.68, P = 0.03). PON1 gene R allele frequency was lower in AD patients than in the controls (60.7% vs 64.7%; chi(2) = 3.85, P = 0.05). One-way ANOVA showed that PON1 genotype had no effect on the age of onset for developing AD. Logistic regression analysis demonstrated the age and sex-adjusted odds ratio (OR) for the risk of AD in PON1 of PON1 R allele carriers was 0.71 (P = 0.044, 95% CI, 0.51 - 0.99).</p><p><b>CONCLUSION</b>Our results indicate that Gln192Arg polymorphism in the PON1 gene is associated with AD, and PON1 R allele might be a protective factor for AD in a Chinese Han ethnic population.</p>

Association between interleukin-1 alpha-889 C/T polymorphism and Alzheimer's disease in Chinese Han population / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the possible association between interleukin-1 alpha-889C/T (IL-1 alpha-889 C/T) polymorphism and Alzheimer's disease (AD) in Chinese Han population.</p><p><b>METHODS</b>A total of 520 AD patients and 505 normal controls were enrolled. The polymorphism of IL-1 alpha-889C/T was detected with real-time polymerase chain reaction. Multiple logistic regression and chi square test were performed for statistical analysis.</p><p><b>RESULTS</b>The frequencies of C/C, C/T, and T/T genotypes were 70.96%, 25.77%, and 3.27%, respectively, among AD patients, and 80.59%, 18.22%, and 1.19%, respectively, among non-dementia controls. In multivariate analysis, T/T and C/T genotypes of IL-1 alpha-889, age > or =65 years, and female were risk factors for AD. Adjusted for the age and sex, T/T and C/T genotypes were still associated with AD. The odds ratio for AD were 3.57 and 1.74 for individuals with T/T and C/T genotypes compared with individuals with C/C genotype. P value was 0. 019 and 0. 001, respectively.</p><p><b>CONCLUSION</b>The IL-1 alpha-889 T/T and C/T genotypes are likely to be susceptible factors for the development of AD in Chinese Han population. The susceptibility genotype, female, and age > or =65 years are risk factors for AD.</p>

Cloning and characterization of a novel mouse aging-related gene Arzc / 中国医学科学院学报

<p><b>OBJECTIVE</b>To clone and identify novel mouse aging-related genes.</p><p><b>METHODS</b>The improved differential display reverse transcription-polymerase chain reaction method was used to analyze the differential expression of old and young BALB/C mouse cortex tissues. The novel expression sequence targets (EST) with differential expression were further confirmed by Northern blot and used as original sequence to clone the full-length cDNA sequence by bioinformatics technique. The reverse transcription products of BALB/C mouse cortex total RNA was used as the amplification template.</p><p><b>RESULTS</b>A total of 42 differentially expressed EST were obtained. Homologic analysis showed that 29 EST represented known cDNAs and 13 were probably new EST. One of the EST, which were confirmed to have down regulative expression in the old mouse cortex tissues by Northern blot analysis, was used to obtain the full-length cDNA sequences from reverse transcription products of total RNA of BALB/C mice cortex by bioinformation method. A 1 382 bp of cDNA sequence including the EST was obtained and designed as Arzc (GenBank accession number: AY344585). The Arzc cDNA contains an open reading frame of 261 bp and encodes a peptide of 86 amino acids residuals. FASTA analysis showed that the deduced peptide had a homology with a portion of integrase/recombinase, a member of a phage integrase family.</p><p><b>CONCLUSION</b>A novel mouse gene that is probably related to aging was identified and cloned.</p>

Association between Alzheimer's disease and nitric oxide synthase III polymorphism / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the association between Alzheimer's disease (AD) and nitric oxide synthase (NOS) III in Chinese Han population.</p><p><b>METHODS</b>Seventy-five AD and 68 normal controls were genotyped for NOS III G894T polymorphism. Genotyping was carried out by PCR-RFLP methods.</p><p><b>RESULTS</b>There were two genotypes of NOS III, GG and GT, in either AD patients or normal controls. The frequencies of these two genotypes were 78.7% and 21.3% in AD patients and 82.4% and 17.6% in normal controls, respectively. No association was found between AD and NOS III genotype (P > 0.05). There were two alleles, G and T, in AD patients and normal controls. The frequencies of these two alleles were 89.3% and 10.7% in AD patients and 91.2% and 8.8% in normal controls, respectively, indicating that there was no association between AD and NOS III allels (P > 0.05).</p><p><b>CONCLUSIONS</b>There was no association between AD and NOS III G894T polymorphism in Chinese Han population.</p>

Identification and analysis of expressed sequence tags related to K562 cells into erythroid differentiation / 中国医学科学院学报

<p><b>OBJECTIVE</b>To isolate expressed sequence tags (ESTs) related to K562 cells erythroid differentiation.</p><p><b>METHODS</b>Modified differential display reverse transcription polymerase chain reaction (DDRT-PCR) method was applied to identify differential ESTs in uninduced and induced K562 cells by HEMIN for 36 hours. Remarkable differential ESTs were firstly selected for cloning, sequencing and bioinformational analyzing. Several ESTs representing new sequence or providing functional clue were selected for Northern blot analysis.</p><p><b>RESULTS</b>Sixty differentially expressed cDNA fragments related to K562 cells inducted into erythroid differentiation by HEMIN were obtained. Among them, 38 were upregulated and 22 downregulated. Among the 40 differential ESTs selected for cloning, sequencing and bioinformationally analyzing, 23 were found to match to known GenBank sequences and 10 represented cDNA sequences with only dbEST database matches and 7 ESTs have no any database matches. The results of 6 in 8 ESTs selected for Northern blot analysis were shown to be consistent with the differential expressions of DDRT-PCR.</p><p><b>CONCLUSIONS</b>The improved DDRT-PCR method had successfully overcome the problem of false positive. These ESTs provide some clue for studying the molecular mechanisms and regulation network of erythroid differentiation.</p>

The correlation between Val66Met polymorphism in BDNF gene and Alzheimer's disease / 中华老年医学杂志

0.05).Our data also showed no significant association between the genotypes and the severity of the disease.One-way ANOVA showed that BDNF genotype had no association to the age of onset for developing AD.Conclusions Our results indicate that Va166Met SNP in BDNF gene is not associated with AD.