Inhibition of invasion and metastasis of human liver cancer HCCLM3 cells by portulacerebroside A.

CONTEXT: Portulacerebroside A (PCA) is a novel cerebroside compound isolated from Portulaca oleracea L. (Portulacaceae), an edible and medicinal plant distributed in the temperate and tropical zones worldwide. OBJECTIVE: This study investigates the effects of PCA in human liver cancer HCCLM3 cells on metastasis and invasion. MATERIALS AND METHODS: After the cells were treated with PCA (2.5, 5, and 10 µg/ml) for 6, 12, 24, or 48 h, adhesion, transwell invasion, and scratch tests were conducted and cell functions were evaluated. Western blot and FQ-RT-PCR assays explored the mechanism of PCA-inhibited invasion and metastasis in the cells. RESULTS: The adhesion rate of the cells was suppressed at 0.5 h (79.4 ± 1.0, 68.7 ± 1.3, and 58.1 ± 1.3%, versus 100 ± 1.5% in the control), 1 h (78.2 ± 1.2, 70.9 ± 1.6, and 55.4 ± 1.9%, versus 100 ± 1.2% in the control), and 1.5 h (71.6 ± 1.1, 62.3 ± 0.9, and 50.4 ± 0.9%, versus 100 ± 1.1% in the control). The 24 h invasion ability was decreased (356.6 ± 11.2, 204.0 ± 17.6, and 113.0 ± 9.5%, versus 443.6 ± 15.4% in the control). The migration capability was also restrained by PCA for 24 h (324.8 ± 25.4, 250.4 ± 21.0, and 126.3 ± 10.1, versus 381.6 ± 30.6 in the control) and 48 h (470.3 ± 34.3, 404.0 ± 19.7, and 201.0 ± 15.4, versus 752.0 ± 63.6 in the control). There was an increase in the mRNA and protein expression levels of TIMP-2 and nm23-H1, inhibition in the mRNA expression of MTA1, MMP-2, and MMP-9, and suppression in the protein expression of MTA1, RhoA, Rac1/Cdc42, MMP-2, but not RhoC and MMP-9. CONCLUSION: PCA suppresses the invasion and metastasis of HCCLM3 cells possibly by modulation of the mRNA and protein expression of related parameters. This is the first study to reveal a new potential therapeutic application of PCA in antimetastatic therapy for liver cancer.

Total Saponin from Root of Actinidia valvata Dunn Inhibits Hepatoma 22 Growth and Metastasis In Vivo by Suppression Angiogenesis.

The root of Actinidia valvata dunn has been widely used in the treatment of hepatocellular carcinoma (HCC), proved to be beneficial for a longer and better life in China. In present work, total saponin from root of Actinidia valvata Dunn (TSAVD) was extracted, and its effects on hepatoma H22-based mouse in vivo were observed. Primarily transplanted hypodermal hepatoma H22-based mice were used to observe TSAVD effect on tumor growth. The microvessel density (MVD), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) are characterized factors of angiogenesis, which were compared between TSAVD-treated and control groups. Antimetastasis effect on experimental pulmonary metastasis hepatoma mice was also observed in the study. The results demonstrated that TSAVD can effectively inhibit HCC growth and metastasis in vivo, inhibit the formation of microvessel, downregulate expressions of VEGF and bFGF, and retrain angiogenesis of hepatoma 22 which could be one of the reasons.

New triterpenoids with cytotoxic activity from actinidia valvata.

Two new triterpenoids, 30-O-beta-D-glucopyranosyloxy-2alpha,3alpha,24-trihydroxyurs-12,18-diene-28-oic acid O-beta-D-glucopyranosyl ester (1) and 2alpha,3beta,3,30-tetrahydroxyurs-12,18-diene-28-oic acid O-beta-D-glucopyranosyl ester (2) were isolated from roots of Actinidia valvata Dunn. Their structures were elucidated by means of extensive spectroscopic studies. Both these two new compounds showed moderate cytotoxic activity in vitro against BEL-7402 and SMMC-7721 tumor cell line.

[Relationships between constitutional types of traditional Chinese medicine and motion sickness in 145 ocean sailors].

OBJECTIVE: To investigate the relationships between constitutional types of traditional Chinese medicine (TCM) and motion sickness. METHODS: A survey of TCM constitutions in ocean sailors participating in a voyage was performed by using the TCM Constitution Questionnaire developed by Beijing University of Traditional Chinese Medicine, while the survey of motion sickness was operated by Graybiel's diagnostic criteria. The incidences of motion sickness among sailors with different types of constitutions were compared. RESULTS: Prior to the voyage, 50.3% of sailors exhibited a gentleness constitution, 14.5% were of dampness-heat constitution, 10.3% were of qi-stagnation constitution, whereas the percentages of qi-deficiency, yang-deficiency, yin-deficiency, blood-stasis and special diathesis constitutions were 6.2%, 7.6%, 6.2%, 4.1% and 0.7%, respectively. None exhibited a phlegm-dampness constitution. By the end of the 176-day voyage, the percentages of gentleness, dampness-heat, qi-depression, qi-deficiency, yang-deficiency, yin-deficiency, blood-stasis, special diathesis and phlegm-dampness constitutions were 33.8%, 13.8%, 13.1%, 11.0%, 6.9%, 9.7%, 4.1%, 0.7% and 6.9%, respectively. The incidence of motion sickness was 69.7% (101 sailors) during this voyage. The incidences of motion sickness among sailors with different types of constitutions before the voyage showed significant difference (P<0.001). The incidence of motion sickness was higher in the sailors with dampness-heat constitution than in those with gentleness constitution. CONCLUSION: Types of Chinese medical constitution can be related to susceptibility to motion sickness. Furthermore, ocean voyage may have an effect or influence on the type of Chinese medical constitution of sailors involved.

[Preventive effects of a sachet of Chinese herbs on influenza and its immune regulation].

OBJECTIVE: To study the clinical efficacy of Pihui Fanggan Sachet (PHFGS), a sachet of traditional Chinese herbs, in preventing influenza and its immune regulation on mice. METHODS: In clinical study, 239 children from Shanghai Baoshan Xubeihong Art Kindergarten were randomly divided into two groups according to different class; 118 children were treated with PHFGS for 45 days as treatment group and 121 children were as blank control. During the observation period, the incidence rate of influenza, the course of disease and the severity of symptoms were recorded. In experimental study, 40 Kunming mice were randomly divided into normal control group (normal mice treated or not treated with PHFGS), and immunocompromised group (immunocompromised mice treated or not treated with PHFGS). Immunocompromise was induced by intraperitoneal injection of 80 mg/kg cyclophosphamide for 3 days. Some mice in the normal control and immunocompromised groups were then treated with extracted solution of PHFGS through nasal cavity for one week. Spleen index, content of CD(3)(+) T cells and CD(4)(+) T cells, CD(4)(+)/CD(8)(+) ratio, activity of natural killer (NK) cells, serum level of interferon γ (INF-γ) and respiratory level of secretory immunoglobulin A (SIgA) were evaluated. RESULTS: The incidence rate of influenza in the treatment group was much lower than that in the blank control group (P<0.01), and the average course was shortened as compared with the control group (P<0.01). The fever, rhinocleisis, runny nose, and throat congestion in the treatment group were improved as compared with the blank control group (P<0.05, P<0.01). After injection of cyclophosphamide, the spleen index, content of CD(3)(+) T cells and CD(4)(+) T cells, CD(4)(+)/CD(8)(+) ratio, activity of NK cells, serum level of INF-γand respiratory level of SIgA in the immunocompromised group were significantly lower than those in the normal control group (P<0.05 or P<0.01), which indicated the immunosuppression. After treated with PHFGS for one week, the spleen index and the respiratory level of SIgA in the immunocompromised group were improved significantly. Although the content of CD(3)(+) T cells and CD(4)(+) T cells, CD(4)(+)/CD(8)(+) ratio and the serum level of INF-γ were improved, the differences did not reach the significant level. No significant effects on immune function of normal mice were observed. CONCLUSION: PHFGS can prevent influenza effectively by improving the immunity, especially the respiratory mucosal immune function.

A rapid and sensitive liquid chromatography-tandem mass spectrometric method for determination of actinoside E in rat plasma and application to a pharmacokinetic study.

A highly sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of actinoside E in rat plasma. The analytes were extracted by ethyl acetate and an analogue of actinoside F was used as the internal standard. The mobile phase consisted of methanol-water (50: 50, V/V) containing 0.1% formic acid was delivered at a flow rate of 0.3 mL·min(-1) to a Zorbax SB-C18 column (100 mm × 2.1 mm, 3.5 µm). The detection was performed by electrospray ionization mass spectrometry in the negative multiple reaction monitoring mode with a chromatograph run time of 3.0 min. Calibration curves of actinoside E were linear in the range of 0.5-2 500 ng·mL(-1). In this range, intra- and inter-day precision ranged from 1.7% to 7.5% and 2.0% to 8.9%, respectively. The accuracy ranged from 95.7% to 108.6%, and extraction recovery from 83.2% to 85.5%. This method was successfully applied to a pharmacokinetic study of actinoside E in rats after intravenous (5 mg·kg(-1)) and oral (100 mg·kg(-1)) administration, and the results showed that actinoside E was poorly absorbed with an absolute bioavailability being approximately 0.27%.

The investigation of the correlation between metabolic syndrome and Chinese medicine constitution types in senior retired military personnel of the People's Liberation Army.

OBJECTIVE: To investigate the incidence of metabolic syndrome (MS) and the distribution of constitutional pattern in elderly retired personnel of the People's Liberation Army. METHODS: Adopting the method of cross-sectional field investigation, from June to December in 2008, the investigation questionnaires were completed by the aged over 60 and collected from 69 military retired residences in the 4 cities of Shanghai, Nanjing, Hangzhou and Qingdao. Other data, including demographic characteristics, physiological characteristics, life style and former medical history, were collected and analyzed. The statistical analysis for the database was drawn up by the software Epidata 3.0. RESULTS: A total of 4,502 people were included in this study, and 35.3% of them were diagnosed with MS. There was no obvious difference in mobility among ages (60 to 69, 70 to 79 and over 80, P>0.05). Referring to the MS patients in the 70s age group, both the phlegm-dampness and dampness-heat constitutional types were evidently higher than those in the 60s age group (P=0.019, P=0.008); while MS patients in 80s and older showed a significantly lower incidence of dampness-heat constitution than those in the 60s (P=0.00); and ql-deficiency constitution was obviously higher in the 80s age group than those in the other two groups (P=0.00). The top 3 constitutions in MS people were, respectively, phlegm-dampness, dampness-heat and qi-deficiency constitution; while in non-MS people, the top 3 constitutions were gentleness, qi-deficiency and phlegm-dampness. When the patient's body mass index (BMI) was more than 25 kg/m(2), the rate of phlegm-dampness and dampness-heat constitution significantly increased, while the rate of qi-deficiency constitution declined; the discrepancy was significant (P=0.00). CONCLUSIONS: The prevalence rate of MS in military senior people was 35.3%, which did not vary among the three age groups. Phlegm-dampness, dampness-heat and qi-deficiency constitution were the three dominant constitutional types seen in the MS patients. The distribution of constitution formation was different in MS people and non-MS people. For different dimensions of BMI, the proportion of each kind of constitutions was varied.

Total saponin from root of Actinidia valvata Dunn prevents the metastasis of human hepatocellular carcinoma cells.

OBJECTIVE: To extract the active component from the root of Actinidia valvata Dunn and to investigate the effects on hepatocellular carcinoma (HCC) cells in vitro. METHODS: Total saponin was extracted from the root of A. valvata (TSAVD). HCC cells, such as BEL-7402, HepG2, PLC, SMMC-7721, MHCC-97-H, and MHCC-97-L, were treated with TSAVD in 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenytetrazolium bromide (MTT) assay. BEL-7402 and MHCC-97-H cells were also treated respectively with TSAVD at different concentrations for 24 h in wound healing and adhesion assays, and the effects of TSAVD on BEL-7402 and MHCC-97-H cells mobility and adhesion abilities were observed. Meanwhile, the effects of TSAVD on invasion and migration of BEL-7402 and MHCC-97-H cells were also investigated by transwell chamber in invasion and migration assays. RESULTS: TSAVD at 1.5 mg/mL inhibited BEL-7402 cell proliferation with inhibition ratios (IRs) of 61.08%, 74.12%, 84.55% at 24, 48, and 72 h, respectively. Meanwhile, TSAVD inhibited MHCC-97-H proliferation in a concentration-dependent manner from 1.5 to 0.5 mg/mL, with the IR of 36% at 1.5 mg/mL at 24 h. For SMMC-7721, PLC, and HepG2, the IR was lower than 30% at 1.5 mg/mL at 24 h. In the wound healing assay, mobility abilities of BEL-7402 and MHCC-97-H cells in TSAVD treated groups were significantly weaker than those of the control group. After pretreatment for 24 h with TSAVD, adhesion abilities were reduced in both MHCC-97-H and BEL-7402 cells, with IRs of 48.50%±4.86% and 49.85%±5.25% at 200 µg/mL. The IRs of MHCC-97-H and BEL-7402 cells in the migration assay were 49.13%±2.91% and 79.37%±0.09% at 200 µg/mL. In the invasion assay, IRs were 69.78%±4.88% and 82.48%±0.25% at 200 µg/mL. CONCLUSIONS: Of all HCC cells, the highest inhibition by TSAVD was seen for BEL-7402 proliferation. TSAVD could restrain adhesion, invasion, mobility, and migration abilities of BEL-7402 and MHCC-97-H cells in vitro.