Monitoring of lung malignant epithelial cells by gene methylation analysis in the conditionally reprogrammed cell cultures.

Conditionally reprogrammed cell (CRC) technology is an effective method for culturing primary malignant cells and non-malignant epithelial cells in vitro. This can be useful for precision medicine applications, such as drug sensitivity assays. However, this approach is commonly hindered by the non-specific growth of non-malignant epithelial cells in CRC cultures and the lack of effective biomarkers/assays to distinguish them from primary tumor cells. In this study, we developed a DNA methylation-based, real-time PCR assay to investigate SHOX2 and PTGER4 gene promoters as sensitive markers for human lung cancer. We first found that in formalin-fixed, paraffin-embedded (FFPE) malignant lung samples, 90% (28/31) had increased SHOX2 and/or PTGER4 promoter methylation as compared with their adjacent non-malignant samples. We then applied this assay to fresh surgical tumors and found increased SHOX2 and/or PTGER4 promoter methylation in 80% (20/25) of tumor samples as compared with their corresponding adjacent non-malignant tissues. Increased methylation of SHOX2 or PTGER4 promoter regions was also detected in 52% (13/25) of CRC cultures. The presence of malignant cells was confirmed by growth in soft agar cultures, a hallmark of malignant transformation, as well by EGFR mutation analysis. These results demonstrate that SHOX2 and PTGER4 promoter methylation levels can be used to detect malignant lung epithelial cells in CRC cultures.

[Studies on the chemical constituents of Rhynchotechum vestitum Hook. F. et Thoms].

Rhynchotechum vestitum Hook, F. et Thoms. mainly distributed in the southern part of Yunnan Province of China, has been used as a folk remedy for the treatment of hepatitis. No information has been found in the literature about its chemical investigation. In this paper, five compounds isolated from the ethanol extract of the stem and root of the plant are reported. Four of them were identified as beta-sitosterol (I), lupeol (II), rubiadin 1-methylether (III) and rubiadin (IV). The fifth, an orange needle crystal (C17H14O6, mp 236, 5-238 degrees C) is a new anthraquinone compound named rhynchotechol. Its structure has been proved to be 1,6-dihydroxy-7,8-dimethoxy-2-methyl-9,10-anthraquinone by spectral analyses and chemical methods.

[Studies on synthesis of baogongteng A--a new myotic agent].

Baogongteng A, isolated from Erycibe obtusifolia Benth., is a new tropane alkaloid used for treating glaucoma. Using synthetic 6 beta-acetoxy-tropinone as starting material, racemic Baogongteng A(8) was synthesized. Synthetic Baogongteng A also shows myotic activities in rabbits, but the potency is half of that of the natural product.