In order to study the stability of akebia saponin D (ASD) in biological fluids in vitro, the determination methods of ASD were established in this study. Akebia saponin D was dissolved in artificial gastric juice, intestinal juice and gastrointestinal contents of rats, respectively, then thermostatically maintained at 37 degrees C. At time intervals after degradation, samples were withdrawn and the concentrations of ASD were determined by HPLC, from which stability of it at different biological specimen was evaluated. As a result, ASD was totally degraded in large intestinal contents of rats in 8 hours. ASD was very stable in artificial gastric juice, intestinal juice and gastric contents of rats. All of the above data proved that ASD was easily degraded by coliform bacteria but stable in acid environment and with the presence of digestive enzyme.
Animals ; Drug Stability ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Gastrointestinal Tract ; chemistry ; metabolism ; Humans ; Rats ; Saponins ; administration & dosage ; chemistry ; pharmacokinetics

Objective To construct a nucleic acid vaccine expressing H.pylori HpaA and inter- leukin-2 gene and to identify the immunogenicity of the vaccine proteins in vitro and protection in vivo. Methods The H paA gene fragment was amplified by polymerase chain reaction(PCR) from the genomic DNA of the standard H.pylori strain 17874.Mouse interlukin(IL)-2 gene was amplified from pClneo- IL-2.The HpaA and IL-2 were cloned into pUCmT vector.After DNA sequences of the amplified HpaA gene and IL-2 were confirmed,both were cloned into the eukaryotic expression vector pIRES through a serial of enzyme digestion and ligation reactions.The recombinant plasmids were screened by PCR and restriction enzyme digestion.Then,recombinant pIRES-HpsA-IL-2 was transfected to COS-7 cells using Lipofectamine~(TM)2000.The immunogenicity of HpaA and IL-2 protein was detected by SDS- PAGE and Western blot.The recombinant plasmids were transformed to LB5000 and then to final host SL7207.The recombinant strains were passaged repeatedly.The mice were challenged with H.pylori after 4 weeks of inoculation of nucleic acid vaccine.H.pylori infection was detected by rapid urease test.Results The amplified HpaA gene fragment and IL-2 were confirmed by sequence analysis.The eukaryotic expression vector plRES and the pIRES-HpaA-IL-2 construction were confirmed by PCR and restriction digestion.The expressions of HpaA(30 000) and IL-2(14 000)protein by pIRES-HpaA-IL- 2 were detected by Western blot.The in vivo study showed that 75.0% and 58.4% of mice vaccinated by HpaA-IL-2 and HpaA,respectively,were protected anaigst H.pylory infection,which was signifi- cant different in comparison with PBS control (P

Objective To research the monoclonal antibody KMP1 inhibited bladder cancer EJ cell lines growth and metastasis in vivo by bioluminescence imaging. Methods Immunohistochemistry was used to determine the KMP1 binding to EJ and EJ-GFP cell lines. The xenograft tumor cell growth and distribution were measured by vernier calipers and dynamic in vivo fluorescence imaging. Immunohistochemistry and H&E counterstaining researched the feature of the xenograft tumor. Results Cell growth curves of EJ and EJ-GFP cells were similar. EJ-GFP had a green fluorescence. In EJ-GFP nude mouse tumor model, the addition of KMP1 significantly inhibited tumor growth and extended the average life span of nude mice. Both EJ and EJ-GFP cells can bind to KMP1,and the weight of transplanted tumors in the KMP1 treatment group was significantly lower than that of the mIgG control group (P＜0.001).Conclusion KMP1 has a promising antitumor effect in vivo. It might be valuable for development as a promising targeted agent for bladder cancer.

We examined the attributed fractions of all-cause, cardiovascular, and respiratory mortality that were attributed to extreme and moderate cold and heat during 2010-2016 in Nanjing. Our results showed that 12.81%, 19.78%, and 25.33% of all-cause, cardiovascular, and respiratory mortalities, respectively, were attributed to temperature. The highest attributed fractions for three types of mortality were at 4 ℃ and the attributed fractions were high around 4 ℃, which falls within the moderate cold temperatures. Although moderate cold has lower RR than extreme cold, it occurred on more days than did extreme cold. Therefore, health burden caused by moderate cold requires further attention in the future.
Cardiovascular Diseases ; mortality ; China ; epidemiology ; Cities ; epidemiology ; Humans ; Mortality ; Respiratory Tract Diseases ; mortality ; Risk Factors ; Temperature

Ligusticum chuanxiong is a well-known traditional Chinese medicine plant. The study on its molecular markers development and germplasm resources is very important. In this study, we obtained 24 422 unigenes by assembling transcriptome sequencing reads of L. chuanxiong root. EST-SSR was detected and 4 073 SSR loci were identified. EST-SSR distribution and characteristic analysis results showed that the mono-nucleotide repeats were the main repeat types, accounting for 41.0%. In addition, the sequences containing SSR were functionally annotated in Gene Ontology (GO) and KEGG pathway and were assigned to 49 GO categories, 242 KEGG pathways, among them 2 201 sequences were annotated against Nr database. By validating 235 EST-SSRs,74 primer pairs were ultimately proved to have high quality amplification. Subsequently, genetic diversity analysis, UPGMA cluster analysis, PCoA analysis and population structure analysis of 34 L. chuanxiong germplasm resources were carried out with 74 primer pairs. In both UPGMA tree and PCoA results, L. chuanxiong resources were clustered into two groups, which are believed to be partial related to their geographical distribution. In this study, EST-SSRs in L. chuanxiong was firstly identified, and newly developed molecular markers would contribute significantly to further genetic diversity study, the purity detection, gene mapping, and molecular breeding.