[Comparative Study on Quality of Volatile Oil of Acori Tatarinowii Rhizoma from Two Habitats].

OBJECTIVE: To compare the quality of volatile oil of Acori Tatarinowii Rhizoma from Guangxi and Sichuan. METHODS: The volatile oil was extracted from Acori Tatarinowii Rhizoma by using steam distillation method, and analyzed by GC-MS. Peak area normalization method was used for calculating the relative percentage contents of chemical constituents, and hierarchical cluster analysis was used for classifying the 20 batches of samples by their relative contents of the main components that were methyleugenol, cis-methylisoeugenol, γ-asarone, ß-asarone and α-asarone. RESULTS: The average extraction rate of 10 batches of volatile oil in Acori Tatarinowii Rhizoma from Guangxi was 1. 61%, and 10 batches of samples from Sichuan was 1. 72%. The relative percentage contents of five main components totaled 78. 19% and 88. 84%, respectively. By t-test, there was no statistical difference between samples from Guangxi and Sichuan. In the hierarchical cluster analysis,10 batches of samples from Guangxi and 10 batches of samples from Sichuan could respectively be classified into four clusters subcategories and five clusters subcategories, while the mean of samples of Guangxi and the mean of samples of Sichuan respectively analyzed with 20 batches of the two habitats that all were classified in the same clusters subcategories. The results of similarity showed that the correlation coefficients of 8 batches in 10 batches of samples from Guangxi were over 0. 9, while 1 batch was only 0. 466. The correlation coefficients of 7 batches in 10 batches of samples from Sichuan had were over 0. 9, while 1 batch was only 0. 069. The correlation coefficients between the mean of samples of Guangxi and the mean of samples of Sichuan was 0. 996. CONCLUSION: Quality of the different batches of volatile oil from Acori Tatarinowii Rhizoma have significant differences, but it has no obvious correlation with the habitats.

Loss of p27 upregulates MnSOD in a STAT3-dependent manner, disrupts intracellular redox activity and enhances cell migration.

Cell migration is a dynamic process that is central to a variety of physiological functions as well as disease pathogenesis. The modulation of cell migration by p27 (officially known as CDKN1B) has been reported, but the exact mechanism(s) whereby p27 interacts with downstream effectors that control cell migration have not been elucidated. By systematically comparing p27(+/+) mouse embryonic fibroblasts (MEFs) with genetically ablated p27(-/-) MEFs using wound-healing, transwell and time-lapse microscopic analyses, we provide direct evidence that p27 inhibits both directional and random cell migration. Identical results were obtained with normal and cancer epithelial cells using complementary knockdown and overexpression approaches. Additional studies revealed that overexpression of manganese superoxide dismutase (MnSOD, officially known as SOD2) and reduced intracellular oxidation played a key role in increased cell migration in p27-deficient cells. Furthermore, we identified signal transducer and activator of transcription 3 (STAT3) as the transcription factor responsible for p27-regulated MnSOD expression, which was further mediated by ERK- and ATF1-dependent transactivation of the cAMP response element (CRE) within the Stat3 promoter. Collectively, our data strongly indicate that p27 plays a crucial negative role in cell migration by inhibiting MnSOD expression in a STAT3-dependent manner.

Kava chalcone, flavokawain A, inhibits urothelial tumorigenesis in the UPII-SV40T transgenic mouse model.

Flavokawain A (FKA) is the predominant chalcone identified from the kava plant. We have previously shown that FKA preferentially inhibits the growth of p53 defective bladder cancer cell lines. Here, we examined whether FKA could inhibit bladder cancer development and progression in vivo in the UPII-SV40T transgenic model that resembles human urothelial cell carcinoma (UCC) with defects in the p53 and the retinoblastoma (Rb) protein pathways. Genotyped UPII-SV40T mice were fed orally with vehicle control (AIN-93M) or FKA (6 g/kg food; 0.6%) for 318 days starting at 28 days of age. More than 64% of the male mice fed with FKA-containing food survived beyond 318 days of age, whereas only about 38% of the male mice fed with vehicle control food survived to that age (P = 0.0383). The mean bladder weights of surviving male transgenic mice with the control diet versus the FKA diet were 234.6 ± 72.5 versus 96.1 ± 69.4 mg (P = 0.0002). FKA was excreted primarily through the urinary tract and concentrated in the urine up to 8.4 µmol/L, averaging about 38 times (males) and 15 times (females) more concentrated than in the plasma (P = 0.0001). FKA treatment inhibited the occurrence of high-grade papillary UCC, a precursor to invasive urothelial cancer, by 42.1%. A decreased expression of Ki67, survivin, and X-linked inhibitor of apoptotic proteins (XIAP) and increased expression of p27 and DR5, and the number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive apoptotic cells were observed in the urothelial tissue of FKA-fed mice. These results suggest a potential of FKA in preventing the recurrence and progression of non-muscle-invasive UCC.

The Chinese herb isolate isorhapontigenin induces apoptosis in human cancer cells by down-regulating overexpression of antiapoptotic protein XIAP.

Although the Chinese herb Gnetum cleistostachyum has been used as a remedy for cancers for hundred years, the active compounds and molecular mechanisms underlying its anti-cancer activity have not been explored. Recently a new derivative of stilbene compound, isorhapontigenin (ISO), was isolated from this Chinese herb. In the present study, we examined the potential of ISO in anti-cancer activity and the mechanisms involved in human cancer cell lines. We found that ISO exhibited significant inhibitory effects on human bladder cancer cell growth that was accompanied by marked apoptotic induction as well as down-regulation of the X-linked inhibitor of apoptosis protein (XIAP). Further studies have shown that ISO down-regulation of XIAP protein expression was only observed in endogenous XIAP, but not in constitutionally exogenously expressed XIAP in the same cells, excluding the possibility of ISO regulating XIAP expression at the level of protein degradation. We also identified that ISO down-regulated XIAP gene transcription via inhibition of Sp1 transactivation. There was no significant effect of ISO on apoptosis and colony formation of cells transfected with exogenous HA-tagged XIAP. Collectively, current studies, for the first time to the best of our knowledge, identify ISO as a major active compound for the anti-cancer activity of G. cleistostachyum by down-regulation of XIAP expression and induction of apoptosis through specific targeting of a SP1 pathway, and cast new light on the treatment of the cancer patients with XIAP overexpression.

Aristolochic acid-associated urothelial cancer in Taiwan.

Aristolochic acid, a potent human carcinogen produced by Aristolochia plants, is associated with urothelial carcinoma of the upper urinary tract (UUC). Following metabolic activation, aristolochic acid reacts with DNA to form aristolactam (AL)-DNA adducts. These lesions concentrate in the renal cortex, where they serve as a sensitive and specific biomarker of exposure, and are found also in the urothelium, where they give rise to a unique mutational signature in the TP53 tumor-suppressor gene. Using AL-DNA adducts and TP53 mutation spectra as biomarkers, we conducted a molecular epidemiologic study of UUC in Taiwan, where the incidence of UUC is the highest reported anywhere in the world and where Aristolochia herbal remedies have been used extensively for many years. Our study involves 151 UUC patients, with 25 patients with renal cell carcinomas serving as a control group. The TP53 mutational signature in patients with UUC, dominated by otherwise rare A:T to T:A transversions, is identical to that observed in UUC associated with Balkan endemic nephropathy, an environmental disease. Prominent TP53 mutational hotspots include the adenine bases of (5')AG (acceptor) splice sites located almost exclusively on the nontranscribed strand. A:T to T:A mutations also were detected at activating positions in the FGFR3 and HRAS oncogenes. AL-DNA adducts were present in the renal cortex of 83% of patients with A:T to T:A mutations in TP53, FGFR3, or HRAS. We conclude that exposure to aristolochic acid contributes significantly to the incidence of UUC in Taiwan, a finding with significant implications for global public health.

[Analysis of the constituents of volatile oil from Fructus Auranti Immaturus by GC-MS].

OBJECTIVE: To analysis the constituents of volatile oil from Fructus Auranti Immaturus by GC-MS. METHODS: The volatile oil was extracted by steam distillation, then separated by capillary gas chromatography. The constituents of volatile oil were identified and their amount were determined by normalization method. RESULTS: Nineteen components were identified from Fructus Auranti Immaturus and their amount accounted 95.791% of total volatile oil. The main components were Limonene (68.25%), gamma-Terpinene (13.02%), alpha-Terpinol (3.28%), beta-Cymene (3.09%), beta-Myrcene (2.34%), alpha-Pinene (1.53%), beta-Pinene (1.05%). CONCLUSION: The main component of volatile oil from Fructus Auranti is Limonene.

[Effects of volatile oil of Rhizoma Acori Tatarinowii on morphology and cell viability in cultured cardiac myocytes].

OBJECTIVE: To observe the effects of volatile oil of Rhizoma Acori Tatarinowii (VOSCP) on morphology and cell viability in cultured neonate rat cardiac myocytes. METHODS: The cardiac myocytes were cultured by trypsin under cool treatment, and the cell purity was assayed with imminocytochemistry; Morphological changes were observed under phase contrast microscope after cardiac myocytes with VOSCP of different concentration for 24h in vitro, and the cell viability was examed by MTT assay. RESULTS: The purity of cultured neonate rat cardiac myocytes was higher than 95%; VOSCP of different concentration could depress pulse frequency and 100-160 mg/L VOSCP can obviously improve the viability of cardiac myocytes. The relation of dose-effect relationship was a parabola whose peak was at the value of 140 mg/L. CONCLUSION: VOSCP of proper concentration can depress pulse frequency and improve the viability of cardiac myocytes.