NMR-based metabolomic approach to understanding Zeng-Sheng-Ping-induced hepatotoxicity, and identifying possible toxic constituents by LC-MS profiles.

Zeng-Sheng-Ping (ZSP) tablets, made from six Chinese herbs, are widely used in the chemoprevention and treatment of precancerous lesions in patients with gastrointestinal cancer. However, sporadic cases of liver injury have occurred. Herein, the serum metabolites in hamsters with ZSP-induced liver damage were analyzed by NMR-based metabolomics. Twelve metabolites associated with ZSP-induced hepatoxicity were identified. Amino acid metabolism and the urea cycle were significantly altered, and three associated amino acid metabolic enzymes (PAH, GS, and GLS) were further validated by ELISA. Therefore, 12 metabolites and 3 amino acid metabolic enzymes were proposed as potential biomarkers in ZSP-induced liver injury. The chemical constituents of ZSP tablets were profiled using liquid chromatography-mass spectrometry. The furanoids in two herbs, Dioscorea bulbifera L. and Dictamnus dasycarpus Turcz., were proposed to be the major hepatotoxic constituents in ZSP, leading to an improved preparation method with low hepatotoxicity for ZSP.

[Diagnostic accuracy of MALDI-TOF mass spectrum in identification of oral Candidiasis isolates].

PURPOSE: To investigate the diagnostic accuracy of matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) in the identification of clinical oral Candida isolates. METHODS: In the first experiment, 170 suspected oral candidiasis patients were included. Unstimulated saliva was collected and cultured on solid plate (Sabouraud's agar) for 48 hours. Clinical isolates were analyzed by both PCR, which was used as golden standard, and MALDI-TOF MS. The diagnostic accuracy of MALDI-TOF MS for both C.albicans and non-C.albicans were investigated. In the second experiment, we tried to identify Candida isolates directly from positive liquid culture in a new cohort of 42 patients. MALDI-TOF MS identification of candida isolates were performed on positive liquid medium cultures directly. The data were analyzed using SPSS 18.0 software package. RESULTS: Totally, 230 clinical Candida isolates were isolated from 212 oral candidiasis patients enrolled. Out of the 230 analyzed clinical isolates, C. albicans (65.65%, 151/230) was the most frequently isolated species, followed by C. glabrata (11.74%, 27/230), and other candida species. The sensitivity and specificity of MALDI-TOF in identification of C. albicans were 93.33% and 92.73% respectively, and those for non-C. albicans were 83.64% and 89.2% respectively. Oral rinse followed by culturing in Sabouraud's liquid medium for 24 hours yield the diagnostic accuracy of 78.42% in MALDI-TOF MS identification. CONCLUSIONS: C. albicans is the most frequent Candida spp. in oral candidiasis. MALDI-TOF MS is an effective and rapid method for identification of Candida spp. The on-plate based method generate much higher diagnostic accuracy than liquid culture based one.

Inhibitory effects of Zengshengping fractions on DMBA-induced buccal pouch carcinogenesis in hamsters.

BACKGROUND: Zengshengping (ZSP) tablets had inhibitory effects on oral precancerous lesions by reducing the incidence of oral cancer. However, the severe liver toxicity caused by systemic administration of ZSP limits the long-term use of this anti-cancer drug. The purpose of this study was to evaluate the tumor inhibitory effects due to the topical application of extracts from ZSP, a Chinese herbal drug, on 7, 12-dimethlbenz(a)anthracene (DMBA) induced oral tumors in hamsters. The study also investigated the anti-cancer mechanisms of the ZSP extracts on oral carcinogenesis. METHODS: DMBA (0.5%) was applied topically to the buccal pouches of Syrian golden hamsters (6 - 8 weeks old) three times per week for six weeks in order to induce the development of oral tumors. Different fractions of ZSP were either applied topically to the oral tumor lesions or fed orally at varying dosages to animals with oral tumors for 18 weeks. Tumor volume was measured by histopathological examination. Tumor cell proliferation was evaluated by counting BrdU labeled cells and by Western blotting for mitogen-activated protein kinase (MAPK) protein levels. The protein levels of apoptosis marker Caspase-3 and regulator Bcl-2 protein were also measured by Western blotting. RESULTS: Topical application of DMBA to the left pouch of hamsters induced oral tumor formation. Animals treated with DMBA showed a loss in body weight while animals treated with ZSP maintained normal body weights. Both the ZSP n-butanol fraction and water fraction significantly reduced tumor volume by 32.6% (P < 0.01) and 22.9% (P < 0.01) respectively. Topical application of ZSP also markedly decreased the BrdU-positive cell numbers in oral tumor lesions and reduced the expression level of MAPK. In addition, ZSP promoted tumor cell apoptosis by increasing Caspase-3 expression but decreasing Bcl-2 protein production. CONCLUSION: The n-butanol and water fractions of ZSP are effective at inhibiting tumor cell proliferation and stimulating apoptosis in oral cancer suggesting that these fractions have chemopreventive effects on DMBA induced oral carcinogenesis.

[Analysis of risk factors for carcinogenesis of oral leukoplakia].

OBJECTIVE: To investigate the risk factors for malignant transformation of oral leukoplakia. METHODS: A total of 409 cases with oral leukoplakia was retrospectively analyzed. Single factor test was first performed to examine the associations between oral leukoplakia's histopathological classification and each of risk factors including sex, age, systemic diseases, course of disease, clinical classification, site, size, numbers of lesion, alcohol and tobacco consumption, and symptom. Then the association of these selected factors with oral leukoplakia's histopathological classification was evaluated using multiple logistic regression analysis. RESULTS: Fifty-two cases of all 409 patients with oral leukoplakia (including 9 severe dysplasia) developed oral cancer. The ratio of malignant transformation was 12.7%. Sex, age, clinical type, site and symptom were chosen as risk factors incorporated into the multiple logistic regression models. The risk of mild-moderate dysplasia in female was 2.40 times as high as that in male. The risk of mild-moderate dysplasia of speckled leukoplakia was 2.81 times as high as that of homogeneous leukoplakia. The risk of mild-moderate dysplasia of dangerous site was 1. 98 times as high as that non-dangerous site. The risk of mild-moderate dysplasia with symptom was 1.84 times as high as that without symptom. The risk of severe dysplasia and oral cancer in female was 3.11 times as high as that in male. The risk of severe dysplasia and oral cancer of speckled (4.50 times), ulcerative (5.63 times), verrucous leukoplakia (4.09 times) were much higher than that of homogeneous leukoplakia. The risk of severe dysplasia and oral cancer in dangerous site was 2.79 times as high as in non-dangerous site. The risk of severe dysplasia and oral cancer in leukoplakia with symptom was 4.38 times as high as without symptom. CONCLUSIONS: The malignant transformation of oral leukoplakia is correlated to sex, clinical type, site and symptom.