Rosmarinic acid (RA), a phenolic acid, is one of the important secondary metabolites produced in Salvia miltiorrhiza. To observe the influence of salicylic acid (SA), an elicitor, on the synthesis of RA and related enzymes, we treated the cell suspension cultures of S. miltiorrhiza with SA and L-a-aminooxy-beta-phenylpropionic acid (AOPP), a competitive inhibitor of tyrosine aminotransferase (TAT). Under this condition, the activities of related enzymes, such as phenylalanine ammonia-lyase and TAT were traced and assayed; the accumulative amount of RA was measured. The results showed that the PAL activity reached the peak at 4 h, 124% higher than that of the control, and the content of RA reached its maximum ((5.914 +/- 0.296) mg/g dry weight) at 8 h, after treated by 6.25 mg/L SA on day 6 of the suspension culture. The results of treatment with 0.1 micromol/L AOPP showed that AOPP affected little on the TAT activity, while the PAL activity was significantly influenced, with 44% lower than that of the control at 6 h. Meanwhile, the reduced accumulation of RA ((4.709 +/- 0.204) mg/g dry weight) paralleled with the decrease in PAL activity. The co-treatment by 0.1 micromol/L AOPP and 6.25 mg/L SA relieved the restriction imposed by AOPP on PAL, and made the cell cultures accumulate more RA than sole treatment with AOPP, indicated that SA induced the accumulation of RA in suspension cell culture of S. miltiorrhiza, and the rate-limiting effect of PAL was stronger than TAT.
Cell Culture Techniques ; methods ; Cinnamates ; metabolism ; Depsides ; metabolism ; Phenylalanine Ammonia-Lyase ; metabolism ; Plant Cells ; metabolism ; Salicylic Acid ; pharmacology ; Salvia miltiorrhiza ; cytology ; growth & development ; metabolism ; Suspensions ; Tyrosine Transaminase ; metabolism

Objective:To investigate the regularity of cervical lymph node metastasis of hypopharyngeal carcinoma and its influence on patient prognosis,to guide the therapeutic strategies for hypopharyngeal carcinoma.Methods:A total of 140 hypopharyngeal squmous cell carcinoma patients who received surgery in Tianjin Medical University Cancer Institute and Hospital from January 2000 to December 2016 were analyzed retrospectively.χ2test was adopted for the analysis of the counting data,and Kaplan-Meier method was used for survival analysis,and the Cox regression model was used to analyze the factors affecting the prognosis.Results:The total rate of lymph node metastasis was 68.6% of the total 140 patients.The rate of occult lymph node metastasis in cN0 patients was 25.0%.The rate of bilateral lymph node metastases was 25%.The most common areas of lymph node metastasis are region II,Ⅲ,andⅣ.The rates of lymph node metastasis were 10.7%,10.4%,7.9% in regionⅠ,ⅤandⅥ,respectively.The total rate of extranodal extension were 66.7%.Univariate analysis showed that cervical lymph node metastasis was closely related to tumor pathological grade(P=0.012),and was not related to gender,age,primary tumor site,tumor T stage(P<0.05).The 3-year and 5-year overall survival rates of patients were 61.0% and 49.1%, respectively.Cox regression analysis showed that the diameter of metastatic lymph nodes(P=0.012),number(P=0.039),and extranodal extension(P=0.010)were significantly related to the survival of patients with hypopharyngeal carcinoma.When the diameter of metastatic lymph nodes≥2.8 cm,number≥2,with extranodal extension,the prognosis is poor.Conclusions:Hypopharyngeal carcinoma has a dismal prognosis,with high rate of lymph node metastasis.Cervical lymph node metastasis is an important factor of the prognosis.The active treatment strategy for the lymph nodes is the key to improve the therapeutic effect of hypopharyngeal carcinoma.

OBJECTIVE: To explore the molecular reasons of weak expression of B antigen on the red cell. METHODS: Serological test for blood group was carried out, including red cell and plasma grouping, and anti-A1 and anti-H testing, and confirming weak A or B antigens by adsorption and elution. Exons 1-7 were sequenced directly, and one of them was cloned and sequenced. RESULTS: All of the 23 samples showed the weak B antigen by serological method. The alleles of the subgroups were identified by DNA sequencing, including 2 Bel subgroup, 4 B3 subgroup, 14 Bw subgroup, 2 CisAB subgroup and a novel allele. The novel allele showed a nucleotide substitution 662G>A in the exon 7, and the sequence was submitted to Blood Group Antigen Gene Mutation Database, and the novel allele was named Bel10. CONCLUSION Nucleotide substitution in exon results in blood subgroup, which showed that the antigens were weakened, and Bw phenotype was the most frequently subgroup.
ABO Blood-Group System/genetics* ; Alleles ; Exons ; Genotype ; Humans ; Nucleotides ; Phenotype