OBJECTIVETo observe the effect of recombinant adenovirus-mediated wild-type p53 gene on the number and proteins of centrosome in K562 cells. To explore the possibility of application of wild-type p53 gene therapy in the treatment of chronic myeloid leukemia.

METHODSThe recombinant adenoviruses carrying wild-type p53 gene (Ad5 wtp53), mutant p53 gene (Ad5 mtp53) or the green fluorescent protein (GFP) gene was repeatedly amplified and co-infected into K562 cells with cation polybrene. The optimal infection titer and infection time of the recombinant adenoviruses were determined by MTT assay, p53 mRNA and protein expression were determined by RT-PCR and Western blot respectively. The centrosomal structural protein gamma-tubulin and the spindle protein alpha-tubulin were marked simultaneously by indirect immunofluorescence staining, and the expression of the centrosomal gamma-tubulin protein, the mitosis and the number of centrosome were observed under the laser confocal microscopy.

RESULTSInfection efficiency with recombinant adenoviruses was facilitated by polybrene in K562 cells, and 4 microg/ml polybrene was chosen. The optimal adenovirus infection titer was 20,000 MOI and the optimal infection time was 72 hours. p53 mRNA and P53 protein can be expressed in K562 cells by Ad5wtp53 and Ad5mtp53. Both the expression of the centrosomal gamma-tubulin protein and the number of centrosomes were decreased after Ad5wtp53 infection.

CONCLUSIONThere is sustained expression of P53 protein in K562 cells after its infection by Ad5wtp53. Wild-type P53 protein can lead to the down-regulation of the number of centrosomes and the expression of centrosomal gamma-tubulin protein in K562 cells.

Adenoviridae ; genetics ; Centrosome ; metabolism ; Genes, p53 ; genetics ; Genetic Vectors ; Humans ; K562 Cells ; Transfection ; Tubulin ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective To explore the activity of brain default mode network (DMN) in patients with mild Alzheimer's disease (AD) by resting state functional magnetic resonance imaging (fMRI) and investigate its possible neural mechanism. Methods Twenty-four patients with mild AD,admitted to our hospital from January 2009 to June 2010,and 25 normal controls (NCs) were chosen in this study.All subjects were examined by Mini Mental State Examination (MMSE), Mattis Dementia Rating Scale (DRS) and fMRI.Resting-state whole brain data were analyzed by amplitude of low frequency fluctuation (ALFF) with two-sample t test and the brain regions in mild AD patients having significantly different ALFF comparing to NCs were observed. Results As compared with that in NCs,the memory function in mild AD patients was seriously impaired (P＜0.05).As compared with NCs,mild AD patients showed significantly decreased ALFF in the posterior cingulate cortex,ventral medial prefrontal cortex and dorsal medial prefrontal cortex, which were closely relevant to the memory (P＜0.05). Conclusion AD patients show significantly decreased active intensity of some DMN nodes that relate to memory in resting state; DMN abnormalities play an important role in early memory impairment of AD patients.

The limitations of antifungal drugs and severe drug resistance make the treatment of invasive fungal infections (IFIs) a great challenge. Itraconazole (ITZ), as a clinical first-line drug, has a wide range of antifungal activity, but it is still limited by adverse reactions such as liver and kidney toxicity, headache and abdominal pain due to its poor water solubility and easy to cause drug accumulation by injection. In this study, the amphiphilic polymer gallic acid-chitosan-cinnamaldehyde (GA-CS-CN) was prepared by amide reaction and Schiff-base reaction. The drug-loaded nanoparticles (GA-CS-CN/ITZ) were prepared by ultrasonic method. The properties of nanoparticles formulations and its in vitro antifungal activity were investigated in the study. Studies have shown that GA-CS-CN/ITZ is spherical, homogeneous and stable, and has good biological safety. The average particle size was 239.57 ± 31.37 nm, ITZ encapsulation efficiency was (93.41 ± 1.12)%, and the cumulative drug release was (62.25 ± 1.88)% at 48 h in vitro. The antifungal activity results of Candida albicans ATCC 10231 (C. albicans) showed that it had the optimal antifungal effect and could significantly enhance the antifungal activity of free drugs. GA-CS-CN/ITZ prepared in this study has excellent biological safety and anti-C. albicans performance, which provides a new choice for the treatment of C. albicans infection and has good application potential in the treatment of this fungal infection.

Beijing has been one of the epicenters attacked most severely by the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) since the first patient was diagnosed in one of the city's hospitals. We now report complete genome sequences of the BJ Group, including four isolates (Isolates BJ01, BJ02, BJ03, and BJ04) of the SARS-CoV. It is remarkable that all members of the BJ Group share a common haplotype, consisting of seven loci that differentiate the group from other isolates published to date. Among 42 substitutions uniquely identified from the BJ group, 32 are non-synonymous changes at the amino acid level. Rooted phylogenetic trees, proposed on the basis of haplotypes and other sequence variations of SARS-CoV isolates from Canada, USA, Singapore, and China, gave rise to different paradigms but positioned the BJ Group, together with the newly discovered GD01 (GD-Ins29) in the same clade, followed by the H-U Group (from Hong Kong to USA) and the H-T Group (from Hong Kong to Toronto), leaving the SP Group (Singapore) more distant. This result appears to suggest a possible transmission path from Guangdong to Beijing/Hong Kong, then to other countries and regions.
Genome, Viral ; Haplotypes ; Humans ; Mutation ; Open Reading Frames ; Phylogeny ; SARS Virus ; genetics

Objective: To analyze the daily drinking behavior and related factors of primary and middle school students in the Nutrition Improvement Program for Rural Compulsory Education Students (NIPRCES) pilot regions. Methods: Multi-stage stratified random cluster sampling method was used to select one to three national pilot counties in 22 provinces in central and western China where the NIPRCES was implemented in 2019. According to different feeding patterns, two primary schools and two middle schools were selected as key monitoring schools. One or two classes were selected from grade 3 to grade 9. The student questionnaire was used to collect the basic information and daily drinking behavior. Taking whether the drinking water ≥5 cups every day as the dependent variable, multivariate logistic regression model was used to analyze the related factors of drinking behavior among students. Results: A total of 27 374 students were included. On average, primary and middle school students in the regions where NIPRCES was implemented had 3.9 cups of water every day. Logistic regression model showed that boys (OR=1.230, P<0.001), primary school students (OR=1.379, P<0.001), father worked outside the home (OR=1.169, P<0.001), both parents worked outside the home (OR=1.228, P<0.001), non-resident students (OR=1.142, P<0.001), the school in the village (OR=1.638, P<0.001) or township (OR=1.358, P<0.001), school feeding (OR=1.252, P<0.001), the school building with flush toilets (OR=1.384, P<0.001) and the central regions (OR=1.300, P<0.001) students were more likely to drink ≥5 cups water every day. Conclusion: The water consumption of primary and middle school students in the pilot regions of NIPRCES is low, and their drinking behaviors are affected by many factors.
Male ; Humans ; Health Behavior ; China ; Drinking Behavior ; Students ; Drinking Water ; Surveys and Questionnaires