OBJECTIVE: To investigate the mechanism of Chinese herbal recipe Weichang'an (WCA) in inducing cell apoptosis of human gastric cancer grafted onto nude mice. METHODS: The high performance liquid chromatography was used for monitoring the stability of WCA. A human gastric cancer cell line SGC-7901 grafted in nude mouse was used as the animal model. The mice were divided into untreated group and two experimental groups. Animals in the two experimental groups received either WCA over a 34-day period or 5-fluorouracil (5-FU) over a 6-day period starting at the 8th day after grafting. Animals in the untreated group received normal saline on an identical schedule. Animals were killed 41 days after being grafted. To assess the effect of the treatment on tumor, the tumor weight was determined by the electron balance immediately after the animals were killed. SP immunohistochemical method was used to detect the expression of proliferating cell nuclear antigen (PCNA) in grafts. Apoptotic indices (AI) of the tumor cells were examined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method. SP method was also used to detect the expressions of cleaved caspase-3, caspase-8 and caspase-9. SYBR green dye I real-time quantitative polymerase chain reaction (real-time quantitative [corrected] PCR) was used to assess the related gene alterations in mRNA level. The expressions of phospho-Stat3 (Tyr705) and bcl-2 proteins were detected by using SP method. RESULTS: Compared with the untreated group, tumor growth was significantly inhibited by treatment of WCA or 5-FU (P<0.01, respectively). The tumor inhibition rate in the WCA-treated group was 48.70% and that in the 5-FU-treated group was 60.10%. The average labeling index (LI) for PCNA in the WCA-treated group and 5-FU-treated group was significantly decreased as compared with that in the untreated group, respectively. The AI of human gastric cancer grafted in the nude mice detected by using TUNEL method was significantly increased to (9.72+/-4.51)% in the WCA-treated group, while it was (2.45+/-1.37)% in the untreated group. 5-FU-treated group was also found a significantly increased AI compared with the untreated group. The expressions of cleaved caspase-3 and caspase-9 in the WCA-treated group and 5-FU-treated group were significantly increased as compared with those in the untreated group. But caspase-8 showed no significant alteration either in the WCA-treated group or in the 5-FU-treated group. The expression levels of Stat3 (2(-)delta delta C(T))=0.16) and bcl-2 (2(-)delta delta C(T))=0.10) detected by using real-time quantitative [corrected] PCR were lower in the WCA-treated group than those in the untreated group. The expressions of phospho-Stat3 (Tyr705) and bcl-2 in the WCA-treated group were significantly decreased as compared with those in the untreated group. CONCLUSIONS: Chinese herbal recipe WCA can inhibit gastric cancer cell SGC-7901 growth in vivo, induce gastric cancer cell apoptosis and suppress the cell proliferation. WCA induces apoptosis through the caspase-9 and caspase-3 pathway in vivo. Its mechanism might be involved in the down-regulation of Stat3 and bcl-2 genes.

OBJECTIVE: To evaluate the growth-inhibiting and anti-metastasis effects of Weichang'an Decoction (WCAD) on orthotopic transplant nude mouse model of human gastric cancer. METHODS: Forty-one nude mice were implanted with SGC-7901 cells at orthotopic site, whereas 25 were implanted with SGC-7901 cells subcutaneously. Then the nude mice in each transplantation model were divided into the same three groups which were WCAD-treated group with WCAD 0.5 ml/d taken orally, 5-FU-treated group with 5-FU 50 mg/kg intraperitoneally injected weekly and untreated control group with physiological saline 0.5 ml/d taken orally. The growth-inhibiting rates of transplant tumors were detected and the metastatic lesions were examined in the orthotopic transplant mouse model while PCNA-positive rate and apoptotic index (AI) were observed in the subcutaneous transplant mouse model. RESULTS: The growth-inhibiting rates in the WCAD-treated and 5-FU-treated groups of orthotopic transplant mouse model were 40.82% and 37.92% respectively whereas those of subcutaneous transplant mouse model were 48.70% and 60.10%. The incidence rates of metastasis in perigastric lymph notes, lymph nodes in the porta hepatis, liver, diaphragm and peritoneum in the WCAD-treated and 5-FU-treated groups were lower than those in the untreated control group, and the total metastasis rates in the WCAD-treated, 5-FU-treated and the untreated control groups were 30.77%, 28.57% and 71.43% respectively with significant differences (P<0.05). The total PCNA-positive rates in the WCAD-treated and 5-FU-treated groups were obviously lower than that in the untreated control group (P<0.05 or P<0.01) while the AI was higher than that in the untreated control group (P<0.01). The growth-inhibiting rate, total PCNA-positive rate and total metastasis rate in the WCAD-treated group had no significant differences as compared with those in the 5-FU-treated group, but the AI in the WCAD-treated group was significantly higher than that in the 5-FU-treated group (P<0.05). CONCLUSION: WCAD has the inhibiting effects on tumor growth and metastasis of gastric cancer which is orthotopic implanted onto nude mice. This effect may be obtained by proliferation suppression and apoptosis induction in cancer.

To evaluate the antiseptic effect of combined using of 5% sodium hypochlorite and calcium silicate?based root canal sealer against Enterococcus faecalis (Ef) biofilms in infected dentinal tubules in vitro . Methods Cells of Ef were inoculated into the dentinal tubules of single?rooted teeth (without caries, periapical lesions and malformations extracted due to periodontal disease or orthodontic reasons; collected from Department of Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University) with centrifugation and incubated in brain?heart infusion (BHI) to form 3?week?old biofilms. The infected samples were subjected to sodium hypochlorite or sterile water bathing for 10 minutes followed by calcium silicate?based root canal sealer (iRoot SP) (calcium silicate?based group), Gutta?percha group and sterile water group placed on the root canal wall for 1, 4 and 12 weeks. There were two samples in each treatment at each point. The antiseptic effectiveness of combined use of sodium hypochlorite and calcium silicate?based root canal sealer was analyzed by laser scanning confocal microscope (LSCM), ANOVA and LSD?t test. Results After treatment with 5% sodium hypochlorite,in calcium silicate?based group for 4 and 12 weeks more Ef biofilm cells [(75.3 ± 3.5)% and (74.8 ± 3.8)% ] were killed than in Gutta?percha group [(65.9±4.1)% and (63.0±3.7)%] and sterile water group [(63.9±4.0)% and (64.2±3.5)%] (P<0.05). After being treated with sterile water, the proportion of dead bacterial cells in calcium silicate?based group for 1, 4 and 12 weeks [(27.5±4.6)%, (43.0±4.4)% and (40.3±6.1)%] were more than those in Gutta?percha group and sterile water group (P<0.05). After being treated with 5% sodium hypochlorite or sterile water, more biofilm bacteria were killed in calcium silicate?based group for 4 and 12 weeks than in calcium silicate?based group for 1 week (P<0.05). Conclusions The combined use of sodium hypochlorite and calcium silicate?based root canal sealer kills more biofilm cells in infected dentinal tubules.

Objective: To evaluate the antiseptic effect of combined using of 5% sodium hypochlorite and calcium silicate-based root canal sealer against Enterococcus faecalis (Ef) biofilms in infected dentinal tubules in vitro. Methods: Cells of Ef were inoculated into the dentinal tubules of single-rooted teeth (without caries, periapical lesions and malformations extracted due to periodontal disease or orthodontic reasons; collected from Department of Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University) with centrifugation and incubated in brain-heart infusion (BHI) to form 3-week-old biofilms. The infected samples were subjected to sodium hypochlorite or sterile water bathing for 10 minutes followed by calcium silicate-based root canal sealer (iRoot SP) (calcium silicate-based group), Gutta-percha group and sterile water group placed on the root canal wall for 1, 4 and 12 weeks. There were two samples in each treatment at each point. The antiseptic effectiveness of combined use of sodium hypochlorite and calcium silicate-based root canal sealer was analyzed by laser scanning confocal microscope (LSCM), ANOVA and LSD-t test. Results: After treatment with 5% sodium hypochlorite, in calcium silicate-based group for 4 and 12 weeks more Ef biofilm cells [(75.3±3.5)% and (74.8±3.8)%] were killed than in Gutta-percha group [(65.9±4.1)% and (63.0±3.7)%] and sterile water group [(63.9±4.0)% and (64.2±3.5)%] (P<0.05). After being treated with sterile water, the proportion of dead bacterial cells in calcium silicate-based group for 1, 4 and 12 weeks [(27.5±4.6)%, (43.0±4.4)% and (40.3±6.1)%] were more than those in Gutta-percha group and sterile water group (P<0.05). After being treated with 5% sodium hypochlorite or sterile water, more biofilm bacteria were killed in calcium silicate-based group for 4 and 12 weeks than in calcium silicate-based group for 1 week (P<0.05). Conclusions The combined use of sodium hypochlorite and calcium silicate-based root canal sealer kills more biofilm cells in infected dentinal tubules.