Effects of fermentation on the hemolytic activity and degradation of Camellia oleifera saponins by Lactobacillus crustorum and Bacillus subtilis.

The saponins, as components of tea seed meal, are undesirable hemolytic components and should be degraded for reducing their hemolytic activity in order to be used in animal feed. In this study, ß-glucuronidase was verified to be a potent hydrolase of tea seed saponins to reduce their hemolytic activity and a ß-glucuronidase-producing Lactobacillus crustorum strain was screened from raw bovine milk. Next, solid-state fermentation with the isolated L. crustorum and a Bacillus subtilis natto strain, which can produce cellulase and hence improve the fermentation performance of tea seed meal, was carried out for detoxification of tea seed meal. The 50% hemolytic dosage (HD50) value of tea seed saponins was increased from 6.69 to 27.43 µg mL-1. The results of LC-MS analysis showed that the percentage of saponin aglycones increased from 30.95 to 84.25% after the fermentation. According to the roles of sugar moieties in hemolytic activity, and the enzymatic hydrolysis characteristics of ß-glucuronidase, the degradation of tea seed saponins from glucosides to aglycones may contribute to the reduction of hemolytic activity. Therefore, tea seed meal may be used as animal feed after fermentation with the tested saponin-degrading microbial strains.

Expression of phosphatase and tensin homolog deleted on chromosome ten in liver of athymic mice with hepatocellular carcinoma and the effect of Fuzheng Jiedu Decoction.

AIM: To explore the expression of phosphatase and tensin homolog deleted on chromosome ten (PTEN) in liver of athymic mice with hepatocellular carcinoma (HCC) and the effect of Fuzheng Jiedu Decoction (FJD). METHODS: Forty eight male BALB/c athymic mice models were built by Bel-7402 with an indirect method. After 24 h of postoperation, the 48 athymic mice were distributed randomly into 4 groups: A, B, C, D, each group had 12 athymic mice. Group A were were treated by intragastric administration with FT207 (Tegafur) for 4 wk. Group B, C and D were treated by intragastric administration with FJD (complex prescription of Chinese crude drug) that had been delegated into 3 kinds of density as the low, middle, and high for 4 wk. At last, athymic mice were put to death, live time, volume of tumors, exponent of tumors and the tumor metastasis in livers were observed; and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry. RESULTS: Four weeks later, the total survival rate in treatment group (A + B + C) was 50% and higher than the control group (0%) treated by FT207, (P < 0.01). The survival rate in group A, B, C was higher than in group D, and except group A with D, there was significant differences (Fisher's Exact Test P = 0.05 or 0.01). And no differences were observed between the treatment groups and the control group in volume of tumors and exponent of tumors (P > 0.05). Tumor metastasis in livers of the treatment group was less than the controls (Fisher's Exact Test, P = 0.021). The result of immunohistochemistry showed that the intensity of PTEN in latero-cancer tissue was the highest, and then the hepatic tissue, the lowest was cancer tissue (Kruskal-Wallis test, chi2 = 60.67, P = 0.000). It also showed that the intensity of PTEN in treatment groups (A, B, C) was higher than the control group (D) (F = 5.90, P = 0.002 in hepatic tissue and F = 15.99, P = 0.000 in latero-cancer tissue and chi2 = 26.08, P = 0.000 in cancer tissue), and group B is the highest in the treatment groups (P < 0.05, r = 0.01. respectively). However, there was no significant statistic difference between group A and group C (P > 0.05). CONCLUSION: FJD can prolong the survival time and decrease tumor metastasis in livers of these experimental mice. Mechanisms of FJD healing HCC may partially be explained by enhancing the expression of PTEN in liver.

[Clinical study of method of strengthening body resistance and disintoxication disintoxication in patients with HCC of post-TACE].

OBJECTIVE: To research the effect of a complex prescription of Chinese crude drug with the function of strengthening body resistance and disintoxication disintoxication in patients with HCC of post-TACE. METHOD: 45 patients with HCC of post-TACE, as the treatment group, were treated by a complex prescription of Chinese crude drug with the function of strengthening body resistance and disintoxication disintoxication and routine methods of protecting liver. Other 37 patients, as the control group, with the same clinical feature were treated by routine methods of protecting liver only. In the later 1 month, accumulated points of clinical symptom, hepatic function and AFP were observed in all of the patients. And the clinical effect of the two groups was compared. RESULT: One week later, in the treatment group, there is no improvement in anorexia but nausea, abdominal distention and lassitude were improved more obviously than pretherapy in both a week and one month later (P < 0.01 or P <0.05). In the control group, anorexia were improved a week later (P <0.05), but there is no improvement in nausea, abdominal distention and lassitude at the same time, and one month later all of the indexes above improved (P <0.01 or P <0.05). Accumulated points of clinical symptom was decreased more obviously in the treatment group than in the control group in both a week and one month later (P <0.05). At the end of the therapy, in the both groups, ALT, TBIL and AFP all improved except ALB, (P <0.01 or P <0.05). And TBIL improved more obviously in the treatment group than in the control one month later (P <0.05). CONCLUSION: This complex prescription of Chinese crude drug can lighten the adverse reaction of post-TACE. And also it can promote the recovery of liver function and evaluate the quality of lives of such patients.

[Expression of PTEN in athymic mice with HCC treated by complex prescription of Chinese crude drug].

OBJECTIVE: To research the treatment effect of complex prescription of Chinese crude drug in BALB/c athymic mice with human liver cancer, which were built by Bel-7402. METHOD: 48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, the 48 athymic mice were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug that had been deliquated into 3 groups by the different density as the low, middle, and high and FT207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision Two-Step Histostaining Reagent). RESULT: All of the 48 athymic mice survived 12 to 28 days (Ms 24 days) and every mouse with liver cancer demonstrated by dissection. The result of immunohistochemistry represents that the intensity of PTEN in latero-cancer tissue is the highest, and then the hepatic tissue, the lowest is cancer tissue, P < 0.01. It also represents that the intensity of PTEN in treatment groups (A, B, C) is more higher than the control group (D), P < 0.05 or P < 0.01, and group B is the highest in the treatment groups, P < 0.05 or P < 0.01. However, there is no significant statistic difference between group A and group C. CONCLUSION: The higher expression of PTEN in the laterocancer tissue can represent the protective reaction of stress of the organism. And anticancer effect of this complex prescription of Chinese crude drug relates to an eligible density of it. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.

[Study of antitumous effect of the complex prescription of Chinese crude drug in athymic mice with HCC].

OBJECTIVE: To research the treatment effect of a complex prescription of Chinese crude drug in BALB/c athymic mice. METHODS: 48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, they were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug of different density as the low, middle, high and FF207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and survial time, volume of tumors, exponent of tumors and the tumor metastasis in livers were observed; and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry. RESULTS: 4 weeks later, the total survival rate in treatment group (A + B + C) was 50% and more higher than the control group (0%) treated by FT207, P < 0.01. The survival rate in group A, B, C was higher than group D, except group A, there was significant differences, P < 0.05 or 0.01. And no differences were observed between the treatment groups and the control group in volume of tumors and exponent of tumors, P > 0.05. Tumor metastasis in livers of the treatment group is less than the control's, P < 0.05. Negative correlation between volume of tumors and survival time, positive correlation between tumors volume and spleen exponent were observed, P < 0.01. The result of immunohistochemistry represented that the intensity of PTEN in latero-cancer tissue was the highest, and then the hepatic tissue, the lowest was cancer tissue, P < 0.01. It also represented that the intensity of PTEN in treatment groups (A, B, C) was more higher than the control group (D), and group B was the highest in the treatment groups, P < 0.05 or P < 0.01. However, there was no significant statistic difference between group A and group C, P > 0.05. CONCLUSION: This complex prescription of Chinese crude drug can prolong the survival time and decrease tumor metastasis in livers of these experimental mice. Too large tumor volume is an main death cause of model. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.