A rapid and high-quality method for total RNA isolation from Haematococcus pluvialis.

Haematococcus pluvialis, as the most potential natural source of astaxanthin, which is a powerful antioxidant with high economic value, has attracted more and more scientific attention in recent years. An in-depth understanding of the mechanism for how H. pluvialis produces astaxanthin requires the intensive investigations on its genetic information. In particular, many reported studies were based on a variety of RNA analyses. However, it is difficult to extract RNA with high quality and quantity from H. pluvialis, because of the blockage from its thick cell wall and contamination by a large quantity of pigments, polysaccharides, and lipids. Therefore, we proposed an optimized Trizol-based RNA extraction method for H. pluvialis by investigating the effect of cell wall broken ways, algal strains, and cell growth status on total RNA isolation. Using this rapid, convenient, and cost-saving method, isolated H. pluvialis RNA had high quantity and quality (with an RNA integrity number of 7.0 and a concentration of 1604.1 ng/µL) equivalent to that isolated by commercial kit, enabling its applications into downstream RNA analyses.

Expression profile of tumor suppressor gene RASSF1 in lacrimal gland carcinoma.

We examined the expression pattern of the tumor sup-pressor gene RAS association domain family gene 1 (RASSF1) in lacri-mal gland carcinoma and analyzed its relationship with the oncogenesis and progression of tumors. Sixty-two patients (30 males, 32 females, average age = 47 ± 3.5 years) admitted with lacrimal gland carcinoma to the Department of Ophthalmology of our hospital between January 2012 and January 2014 were enrolled in this study. Based on tumor ma-lignancy, patients were classified into a malignant group (N = 25) and benign group (N = 37). Healthy lacrimal gland resections from trauma surgery (N = 35) were recruited as a healthy control group. Expres-sion profiles of RASSF1 in all groups were quantified using reverse transcription-polymerase chain reaction and western blotting. Recur-rence of lacrimal gland carcinoma was surveyed through postopera-tive follow-up. Expression levels of RASSF1 in samples from the ma-lignant and benign groups were significantly lower than those in the healthy group (P < 0.05). Furthermore, the malignant group showed lower RASSF1 expression than the benign group (P < 0.05). Postopera-tive follow-up identified 22 cases of recurrence in the malignant group, with a recurrence rate of 88%, while 15 cases in the benign group had a recurrence rate of 40.5%. A direct relationship exists between RASSF1 expression levels and the malignancy grade of lacrimal gland carci-noma. Patients with lower RASSF1 expression showed a higher recur-rence probability, indicating unfavorable prognosis. Therefore, measur-ing RASSF1 expression can be used as a diagnostic method for lacrimal gland carcinoma.