Objective To upregulate Notch signaling in cancer cells by overexpression of active part of Notch1 and to examine the proliferation of the cells. Methods Four cancer cell lines were infected with retrovirus recombined with sequence encoding active part of Notch1.CBF-1 reporter plasmid was used to detect Notch signaling and proliferation assay was carried out by MTS method.Cell cycle analysis was synchronously conducted. Results The overexpression of the active part of Notch1 induced upregulation of Notch signaling,led to growth inhibition in Hela and HepG2 cell lines and growth boost in BGC-823 cell lines,while had no effect on Chang cell lines. Conclusion The upregulation of Notch signaling can exert various effects on different cancer cell lines which is critical to the gene therapy for cancers.

Objective To learn the current situation,effect and the matters of the internet class teaching quality assessment system and to propose some suggestions and measures for teaching quality assessment scientifically.Methods We made a questionnaire among the undergraduates of the Chongqing Medical University,using Epidata3.02 for data recording and SAS8.2 for data analysis.Result The accessibility of the class teaching quality net assessment system was not very ideal,and most students were not familiar with the pathway for assessing the teaching quality on internet;the students were generally objective and fair in the teaching quality assessment,but they were not very energetic and initiative;the students had the desire to communicate,but the teachers did not think highly of the teaching assessment;the current assessment index and the internet class teaching quality assessment system should be improved.Conclusion We should emphasize the publication and education of the internet class teaching quality assessment system,raise the recognition of both teachers and students on the role and significance of the internet assessment system,upgrade and improve the assessment index system and promote the close association of the assessment and the actual teaching status.

Age Factors ; Child Nutritional Physiological Phenomena ; Child, Preschool ; Diet ; Eating ; Female ; Humans ; Male ; Sex Factors

In recent years, benefited from the progress of clinical studies of early diagnosis and early revascularization in acute myocardial infarction (AMI), the guidelines for Europe and the United States and the Chinese academic continue to be updated to guide our clinical practice. However, AMI still remains one of the major causes of death in the worldwide. Over the past 10 years, the incidence of AMI increased rapidly in China, and the mortality kept at a high level. There is still plenty of room to improve in the early diagnosis of cardiac troponin, revascularization strategy optimization of non-infarct-related vascular, optimized new antiplatelet therapy, development of regional synergies network and chest pain center. There is still a long way from the standardized treatment of AMI. It is very important to complete early diagnosis and optimum treatment of AMI.

Objective To analyze clinical characteristics of invasive Luminal subtype breast cancer.Methods The data of 162 invasive Luminal subtype breast cancer patients receiving operation in Cancer Hospital of Chinese Academy of Medical Science from January 1 st to September 30th in 2002,were collected and the clinical characteristics,recurrences,metastasis and survivals were retrospectively analyzed.Results The median time of follow-up was 92 months,ranging from 4 to 98 months.41 cases (25.3％,41/162) presented local recurrence or metastasis including 32 cases with metastasis ( 19.8％,32/162),2 cases with local recurrences (1.2％,2/162) and 7 cases with both local recurrence and metastasis (4.3％,7/162) ;Disease-free survival (DFS) and the 5-year DFS were 73.1％ and 79.6％,respectively.27 patients ( 16.7％,27/162) died of breast cancer,the overall survival (OS) and 5-year OS were 82.5％ and 85.3％,respectively.According to Kaplen-Meier survival analysis,tumor size,lymph node status and clinical stage were correlated to overall survival time ( P ＜ 0.05 ) ; and rumor size,lymph node starus,grade,clinical stage and PR status were correlated to disease-free survival time ( P ＜ 0.05 ).By multivariate analysis,TNM stage,PR and PCNA were independent prognostic factors correlated to overall survival time (OR=0.633,95％ CI:0.411 -0.976,P＜0.05; OR =0.823,95％ CI:1.012-3.283,P ＜ 0.05) ; TNM stage and PR was independent prognostic factors correlated to disease-free survival time (OR =3.273,95％ CI:1.719 - 6.232,P ＜ 0.01 ; OR =0.599,95％ CI:0.423 - 0.850,P ＜ 0.01 ).Conclusions In invasive Luminal subtype breast cancers,PR is correlated to fine prognosis,and PCNA is correlated to overall survival time.

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1,5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.

Objective To investigate the influence of temperature on active constituents of the superfines of Qianghuo (Notopterygium). Methods Qianghuo was micronized by using vibromill at room temperature and low temperature (-10 ℃) respectively. The content of volatile oil in the superfines prepared at different temperature was detected, and the content of ferulic acid was determined by using HPLC.Results The content of ferulic acid was not lost after micronized by vibromill at room temperature, but the content of volatile oil was lost, which could be avoided by the micronization at low temperature (-10 ℃). Conclusion The vibromill at low temperature is proved initially that it is suitable for the preparation of superfines of Qianghuo.

Objective To investigate the effect of Porphyromonas gingivalis(Pg) with different fimA genotypes on vascular cell adhesion molecule-1(VCAM-1) and intercellular adhesion molecule-1(ICAM-1) production by human umbilical vein endothelial cells(HUVEC). Methods In the present study, PgATCC33277(type Ⅰ fimA genotype), WCSP 115(type Ⅱ fimA genotype), W83(type Ⅳ fimA genotype), and Escherichia coli-lipopolysaccharide (Ec-LPS) were designed as experimental group 1, 2, 3, and positive control group, respectively, to stimulate HUVEC, and the un-stimulated HUVEC were analyzed as negative control group. The three strains of Pg were cultured anaerobically in standard condition, and then the Pg cells and Ec-LPS were co-cultured with HUVEC for 2, 6, and 24 h, respectively. The amount of ICAM-1 and VCAM-1 produced by HUVEC was detected with flow cytometry(FCM). The expression of ICAM-1 and VCAM-1 by HUVEC were assayed with confocal laser scanning microscope(CLSM). ResultsThe expression of ICAM-1 on the surface of HUVEC were intensified after infected by Pg with Ⅰ, Ⅱ, and Ⅳ fimA genotypes (P<0.05). The amounts of ICAM-1 were 60.27±5.43, 80.81±1.44, and 85.94±2.56 for Pg with type Ⅰ fimA genotype, 86.69±8.81, 90.19±0.00, and 96.18±0.48 for Pg with type Ⅱ fimA genotype, 59.66±0.40, 85.79±4.86, and 96.04±2.07 for Pg with type Ⅳ fimA genotype at 2, 6 and 24 h after infection, respectively. The up-regulation effects caused by Pg with type Ⅱ and Ⅳ fimA genotypes were stronger than those caused by Pg with type Ⅰ fimA genotype at different time points except at 2 h(P<0.05). Under the present experimental condition, infected by Pg with type Ⅰ, Ⅱ and Ⅳ fimA genotypes stimulated low expression of VCAM-1 by HUVEC, it showed no significant differences among all the groups (P>0.05). Expression of ICAM-1 and VCAM-1 in Pg infected HUVEC were confirmed by CLSM. Infection of HUVEC with Pg resulted in more fluorescence staining of ICAM-1 and VCAM-1 compared with that in uninfected HUVEC cultures. Conclusions The virulence and pathogenicity of Pg is associated with its fimA genotypes, Pg with type Ⅱ and Ⅳ fimA genes possess stronger ability to stimulate HUVEC to up-regulate the expression of cell adhesion molecules, which may lead to disorders in vascular endothelial function.

LTB gene fragment was amplified by PCR from plasmid pMDTLT, and a recombinant plasmid pETLTBVP1 was constructed by inserting LTB gene fragment into VP1 gene expression plasmid pETVP1 constructed previously. The recombinant plasmids were transformed into E. coli BL21(DE3) and induced to express by IPTG. The recombinant protein existed in the inclusion body and its molecular weight was about 39 kD proved by SDS-PAGE analysis. Western blotting showed that the fusion protein could be reacted with both anti-FMDV and anti-cholera toxin serum demonstrating the immunoactivity of the fusion protein. Strong immune responses can be induced in mice inoculated with the fusion protein intraperitoneally, and the serum antibody level is higher than that of commercial foot-and-mouth disease vaccines.
Animals ; Antibodies, Viral ; blood ; Bacterial Toxins ; genetics ; immunology ; metabolism ; Capsid Proteins ; genetics ; immunology ; metabolism ; Enterotoxins ; genetics ; immunology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Escherichia coli Proteins ; Female ; Gene Fusion ; genetics ; Mice ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

This study is to compare the effects of kaempferide and anhydroicaritin on biomineralization of rat osteoblasts (ROB) in vitro. Calvarias were dissected aseptically from newborn SD rats, the osteoblasts were obtained by enzyme digestion and were cultured in MEM containing 10% FBS. The medium was changed every three days, and serial subculture was performed when cells covered with 90% of the dish. Kaempferide and anhydroicaritin were separately added with final concentrations of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) and 1 x 10(-7) mol x L(-1) under the conditions of osteogenic differentiation. The proliferation was measured by MTT, and the optimal concentration was detected by the ALP activity at the 9th day after osteogenic induction culture. The osteogenic indexes of kaempferide, anhydroicaritin and control group with the optimal concentration were compared. The result showed that the anhydroicaritin at concentration of 1 x 10(-5) mol x L(-1) had significantly promoted the activity of ALP, calcium content and osteocalcin content, increased the number of CFU-F(ALP) and mineralized nodules, enhanced the mRNA level of BMP-2, OSX and Runx-2, which are key genes of osteogenic differentiation, and raised the protein content of collagen-I. However, the kaempferide group had not significantly represented the ability that promoted osteogenic differentiation of ROB. The difference of osteogenic differentiation on ROB between kaempferide and anhydroicaritin was caused by the prenyl group on C-8 of icariin.
Alkaline Phosphatase ; metabolism ; Animals ; Benzopyrans ; pharmacology ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Calcium ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; metabolism ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Kaempferols ; pharmacology ; Osteoblasts ; cytology ; metabolism ; Osteocalcin ; metabolism ; Osteogenesis ; drug effects ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transcription Factors ; genetics ; metabolism