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1.  Factors Influencing the Abundance of the Side Population in a Human Myeloma Cell Line 
Bone Marrow Research  2011;2011:524845.
Side population (SP) refers to a group of cells, which is capable to efflux Hoechst 33342, a DNA-binding dye. SP cells exist both in normal and tumor tissues. Although SP abundance has been used as an indicator for disease prognostic and drug screening in many research projects, few studies have systematically examined the factors influencing SP analysis. In this study we aim to develop a more thorough understanding of the multiple factors involved in SP analysis including Hoechst 33342 staining and cell culture. RPMI-8226, a high SP percentage (SP%) human myeloma cell line was employed here. The results showed that SP% was subject to staining conditions including: viable cell proportion, dye concentration, staining cell density, incubation duration, staining volume, and mix interval. In addition, SP% was highest in day one after passage, while dropped steadily over time. This study shows that both staining conditions and culture duration can significantly affect SP%. In this case, any conclusions based on SP% should be interpreted cautiously. The relation between culture duration and SP% suggests that the incidence of SP cells may be related to cell proliferation and cell cycle phase. Maintaining these technical variables consistently is essential in SP research.
PMCID: PMC3196862  PMID: 22046565
2.  Quantitative analysis of two isoflavones in Pueraria lobata flowers from eleven Chinese provinces using high performance liquid chromatography 
Chinese Medicine  2010;5:14.
Pueraria lobata flower (Gehua) is a medicinal herb to treat intoxication, hepatic and gastrointestinal tract lesion induced by alcohol. This study aims to develop a new HPLC method for the determination of two major isoflavones in P. lobata flowers, namely tectoridin and 6"-O-xylosyl-tectoridin.
A high performance liquid chromatography (HPLC) method with a C18 column (250 mm × 4.6 mm, 5 μm) was developed for the quantitative analysis of tectoridin and 6"-O-xylosyl-tectoridin, the main isoflavone components in P. lobata flower. A simple gradient of acetonitrile/water (0 min 15:85; 35 min 50:50; 36 min 15:85; 40 min 15:85; v/v) was used, and 265 nm was selected as detection wavelength. Tectoridin and 6"-O-xylosyl-tectoridin were used as the external standards in quality control of P. lobata flower for the first time. The method was applied to practical use in quality assessment of eleven batches of P. lobata flower samples in Chinese herbal medicine market.
The peak area response was linear for tectoridin in the 11.8-236.4 μg/mL range with a correlation coefficient of 0.9996 (P < 0.001), and for 6"-O-xylosyl-tectoridin in the 10.33-185.99 μg/mL range with a correlation coefficient of 0.9984 (P < 0.001) respectively. The average recoveries were 102.7-103.7% for tectoridin and 95.7-103.2% for 6"-O-xylosyl-tectoridin (RSDs < 3%), and the intra-day and inter-day RSDs of the two components were less than 2%. This HPLC method was applied to assess the quality of P. lobata flower from eleven provinces in China. P. lobata flowers from northern China contained 26.46-43.28 mg/g of tectoridin and 30.90-48.23 mg/g of 6"-O-xylosyl-tectoridin comparing to 10.00-19.81 mg/g of tectoridin and 11.08-37.03 mg/g of 6"-O-xylosyl-tectoridin in those from southern China.
The results showed that P. lobata flowers from northern China contained more tectoridin and 6"-O-xylosyl-tectoridin than those from southern China.
PMCID: PMC2876171  PMID: 20416072

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