Aim To observe the effect of methotrexate (MTX) on proliferation, migration and apoptosis of cultured vascular smooth muscle cells (VSMC). Methods Rabbit thoracaortic VSMC were cultured in vitro.VSMC proliferation was evaluated by cell counting and cell cycle analysis. Monolayer cell scrape was used to observe VSMC migration. Apoptosis was observed with flow cytometry, DNA gel electrophoresis and TUNEL stain. Results MTX (25～100 nmol?L -1) inhibited VSMC proliferation in a dose-dependent manner.25 nmol?L -1 and 50 nmol?L -1 MTX increased the percentage of the S phase cells and decreased the percentage of the G 2/M phase cells (P

Objective:To explore the effects of "2+1" PAD class based on online course in medical statistics undergraduate teaching.Methods:A total of 326 clinical undergraduates who took medical statistics in the spring term of 2017 were randomly divided into experimental group ( n=186) and control group ( n=140). Students in the experimental group were taught by "2+1" teaching method, which included self-study on line, classroom teaching, discussion and so on. While students in the control group were taught traditionally. Feedbacks about the teaching progress and effects from the students in the two groups were compared. SAS 9.4 was used for quantitative data description, t-test and rank sum test. Results:according to the students, the online-based "2+1" teaching method paid more attention to interdisciplinarity than traditional method ( P<0.01), and the new teaching method was more beneficial for the improvement of students' capacity ( P=0.008) and more conducive for students to engaging in scientific research ( P=0.012). Students in the experimental group had higher overall satisfaction scores ( P<0.01) and better ability for solving comprehensive problems ( P<0.01). Conclusion:The "2+1" PAD class teaching model based on online courses has more advantages than traditional teaching methods does, which should be recommended and adopted in undergraduate teaching.

Objective:To prepare the virus-like particle (VLP) of the GII.3[P12] human norovirus (HuNoV) strain GZ2013-L20 in Guangzhou and its polyclonal antibody, and systematically characterize its immunogenicity and receptor binding ability, which would provide data for prevention and control of HuNoV.Methods:ORF2 gene was amplified from the genome of the GZ2013-L20 strain to construct the recombinant transposon vector, which was further transformed into Escherichia coli DH10 Bac to develop the recombinant baculovirus Bacmid-L20-ORF2. VLP was expressed in the sf9 insect cells and then purified. Transmission electron microscopy, SDS-PAGE, Western blot (WB), and receptor binding experiments were performed to characterize the purified VLP. In addition, the polyclonal antibody from the immunized mice was evaluated by indirect enzyme-linked immunosorbent assay (ELISA) and the blocking test of receptor binding. Results:The recombinant baculovirus plasmid Bacmid-L20-ORF2 was constructed, and the target VLP was successfully obtained. The result by the transmission electron microscope demonstrated that the VLP were about 30 nm in diameter. SDS-PAGE and WB analyses showed that the protein’s relative molecular mass (Mr. ×10 3) was about 58. The result of receptor binding experiments showed that the VLP could bind to the secretory salivary receptors (types of A, B, AB and O), non-secretory salivary receptors (O type) and the porcine gastric mucin. The polyclonal antibody with a titer of 2 × 10 5 was detected in the immunized mice, which showed strong cross-immunoreactivity with capsid proteins of 20 (20/28) HuNoV genotypes. In addition, the result of blocking tests of receptor binding showed that the VLP polyclonal antibody only blocked the viral VLP of the same genotype, but had no neutralizing effects on the VLPs of GII.2, GII.4, GII.8 and GII.17. Conclusions:The VLP of GII.3[P12] HuNoV Guangzhou strain showed strong binding ability to both secretory and non-secretory salivary receptors, and its polyclonal antibody showed a broad spectrum of immunobinding, but its neutralization blocking feature was effective only against the virus of the same genotype. The result provide basic data for rational design of vaccine development.

Objective To investigate the association of silent information regulator 1 (SIRT1) polymorphisms with intracerebral hemorrhage (ICH) susceptibility.Methods From September 1,2013 to May 30,2017,Han ethnic 201 ICH patients and 203 controls from South China were enrolled in this study.Genotyping and sequencing ofSIRT1 polymorphisms (rs7069102,rs2273773 and rs7895833) were performed by PCR-restriction fragment length polymorphism (PCR-RFLP).The correlation of SIRT1 polymorphisms with ICH was analyzed.Results (1) The rs7895833 A allele frequency distribution was significantly higher and the rs7895833 GG+AG gene frequency distribution was significantly lower in the ICH group than those in the control group (P＜0.05);the rs7069102 C allele frequency distribution was lower and the GG+CG gene frequency distribution was higher in the ICH group than those in the control group,without significant differences (P＞0.05).(2) Logistic regression analysis indicated rs7895833 AA genotype carriers had increased risk for ICH (OR:1.57,95％CI:1.14-2.18,P=0.006).(3) As compared with patients with rs2273773 TT genotype,patients with CC and CT genotypes had significantly higher high-density lipoprotein cholesterol level (P＜0.05);there were no associations between rs2273773/rs7069102 and ICH.Conclusion SIRT1 rs7895833 is significantly associated with ICH susceptibility;rs2273773 genotypes affect plasma high-density lipoprotein cholesterol level in the Chinese Han ethnic population.