High-quality RNA isolation from recalcitrant adipose tissue with high lipid content and low cell numbers is difficult. Many studies have made efforts to optimize methods for isolating RNA from adip... Method summary A total of 22 human orbital fat specimens of the same quality were split into two groups, classic and optimized. RNA were extracted by the traditional method and the optimized method... Tweetable abstract An optimized protocol based on TRIzol for RNA extraction from human fat is described. Graphic abstract Optimized protocol yielding RNA from lipid-rich specimens.

Retinoblastoma (RB) is a primary intraocular malignancy in childhood, and may develop relapse and metastatic disease. This study was to identify the stem-cell properties of primary retinoblastoma cells critical to tumorigenesis and metastasis. Primary cells were isolated from fresh human RB tissues after enucleation, and cultured in serum-free or serum-enriched conditions, with two RB cell-lines Weri-RB1 and Y79 for comparison. Proliferation of primary RB cells were well-maintained in serum-free condition of DMEM/F12 medium, and formed stem-cell like spheroids. The immaturity of cultured primary RB cells was demonstrated by tendency of highly expressed stem-cell markers (CD133, Nestin and OCT4) and suppressed mature retinal-cell markers (GFAP, MAP2 and Recoverin). CD133, a neural stem-cell marker being exclusively studied in RB, was found positive in small patches of cells in archival human RB by immunohistochemistry. Meanwhile, at initial isolation, insignificant CD133+ cells were detected by flow-cytometry, and substantial increase of positivity was observed after several days cultivated in serum-free condition. Cultured primary RB cells were engrafted in subretinal region of BALB/c nude mice for assessment of tumorigenicity. Strong tumorigenic activity and extensive progression of the xenograft retinoblastoma was induced by primary cells as compared with the two cell-lines. Again, immunohistochemistry confirmed that the stem-cell markers were emphasized in the xenograft tumor in mice. Our findings demonstrated that in comparison to the well-established RB cell-lines, cultured primary RB cells possess stem-cell like properties with highly expressed stem-cell markers, self-regenerative growth in culture, and strong in vivo oncogenic potentiality.

Purpose: The periocular features vary in different age groups and ethnic populations. In this study, the authors sought to determine whether age-related differences exist for certain periocular parameters between children and young adults of Chinese Han population. Methods: OU of 310 children (aged 7–10 years) and 301 young adults (aged 20–30 years) were evaluated by 2-dimensional photogrammetry. All the subjects were of Chinese Han population. The eyelid crease profiles were classified according to their morphology. Periocular measurements, including 11 linear and 3 angular parameters were acquired from standardized photographs, 3 indices were further derived. The presence of epicanthal fold was also recorded. Each of the parameters was compared between the age groups. Results: A statistical difference was found between the 2 age groups for the distribution of eyelid crease types. Statistical significances existed between the age groups for various parameters, these include palpebral fissure height and width, margin reflex distance, intercanthal and outercanthal width, distance from the medial end of the eyebrow to the medial canthus, distance from the lateral end of the brow to the lateral canthus, crease height and eyebrow height, angle of endocanthion and exocanthion, palpebral fissure inclination, palpebrae fissure index, canthal index, and angular index (p < 0.05). The incidence of epicanthal fold tended to decrease with increase of age. Conclusions: Age differences exist in periocular features between children and young adults of Chinese Han population. Standards of periocular measurements based on age and race could provide reliable guidelines for clinical examination and planning periocular surgery.