ABSTRACT
Objective:To investigate the targets and possible mechanism of Didangtang in the treatment of bladder cancer. Method:Based on multiple traditional Chinese medicine and disease databases, the network pharmacology was used to screen potential targets, analyze the biological functions of potential targets, and construct a network of "Chinese medicine-target-path-disease". Bioinformatics analysis was applied in population and gene databases, in order to explore the differential expressions of core targets in tissues, distribution in the population and the correlation with prognosis. The in vitro experiment was used to verify the biological function of Didangtang. The underlying mechanism of Didangtang on the candidate target was detected. Result:A total of 21 core target genes and 16 highly enriched pathways were screened out. A functional network of Didangtang was constructed systematically. At the same time, six targets, namely cadherin 1 (CDH1), CAMP responsive element binding protein 1 (CREB1), colony stimulating factor 2 (CSF2), AP-1 transcription factor (JUN), matrix metalloproteinase 2 (MMP2), and prostaglandin-endoperoxide synthase (PTGS2), were differentially expressed in bladder cancer tissues (P<0.05). Furthermore, JUN and MMP2 were also differentially distributed in population (P<0.05). At the same time, the expression level of JUN was correlated with the prognosis of patients with bladder cancer (P<0.05). The in vitro experiment revealed that Didangtang inhibited the proliferation of bladder cancer cells and decreased the expression of candidate target JUN (P<0.01). Conclusion:Didangtang has the characteristics of multiple targets and multiple pathways in treatment of bladder cancer. It is initially confirmed that Didangtang can affect the expression of target JUN and inhibit the proliferation of bladder cancer, which lays a good foundation for further studies on mechanism.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of exposure to vehicle exhaust in pregnant mice on the reproductive function and DNA methylation in male offspring mice.</p><p><b>METHODS</b>Twenty pregnant mice were randomized into control group and vehicle exhaust exposure group (n=10) and exposed to routine laboratory condition and to vehicle exhaust for 10 consecutive days (8 h per day) in a tunnel with a heavy traffic, where the concentrations of TSP, PM10, PM2.5, SO2 and NOX and the decibel of noise were measured. The offspring mice were raised till reaching maturity, and the epididymides of the male mice were collected to test the weight coefficients, DNA methylation level, and mRNA levels of Aldh7a1 and Rpe.</p><p><b>RESULTS</b>The body weight and the weight coefficients of the epididymides and testes differed significantly between the exposure group and the control group (P>0.05). The concentrations of TSP, PM2.5, PM10 and NOx and the decibel of noise were significantly higher in the traffic environment and the control environment (P<0.05). Reduced representation bisulphite sequencing (RRBS) and Gene ontology (GO) showed that 58 genes had significantly different methylation levels between the two groups, mostly relating to the process of spermatogenesis (P<0.05). Compared with the control group, Aldh7a1 and Rpe mRNA expressions in the testes were down-regulated significantly in the exposure group (P<0.05).</p><p><b>CONCLUSION</b>Exposure of pregnant mice to vehicle exhaust causes damages of the reproductive function in the male offspring mice.</p>
ABSTRACT
The M protein gene of porcine reproductive and respiratory syndrome virus amplified by PCR was tandem linked with its GP5 gene in shuttle vector in correct frame, resulting in shuttle vector pShuttle-CMV-M-GP5. The positive clone was identified by PCR and further confirmed by sequencing. The constructed plasmid was linearized with Pme I and co-transformed BJ5183 host bacteria with pAdEasy-1 to produce recombinant adenovirus DNA by homologous recombination. Then the adenovirus DNA was linearized with Pac I and transfected into HEK-293A cells to obtain recombinant adenovirus. The specific expression of target proteins by the recombinant adenovirus was verified by indirect immuno-fluorescence assay (IFA) with monoclonal antibodies against M and GP5.The results showed that the tandem linked M with GP5 could be co-expressed by adenovirus vector. Mice immunized with the constructed recombinant adenovirus induced strong humoral immunity (ELISA antibody and virus neutralizing antibody) and cellular immunity (lymphocyte proliferation and CTL responses). The results showed that the recombinant adenovirus has strong immunogenicity and provided the basis for the further experiments in pigs.