ABSTRACT
OBJECTIVE@#To establish a culture system for human nasal mucosal organoids with controllable differentiation to reproduce the structure and function of the source tissue through staged expansion-differentiation culture.@*METHODS@#Fresh samples of surgically resected middle turbinate and nasal polyp tissues were collected, from which the nasal mucosa epithelial cells were isolated by enzymatic digestion and filtration for continuous culture at the air-liquid interface for expansion (EO group) or staged culture for expansion and differentiation (DO group). Immunohistochemical staining was used to characterize the structure, cellular composition and ciliary function of nasal mucosal organoids in the two groups. The secretion function of the differentiated nasal mucosal organoids in DO group was evaluated using PAS staining.@*RESULTS@#Both of the two organoid culture systems yielded vacuolar or solid spherical 3D organoids, and their diameters increased progressively with time. On day 16 of culture, more vacuolar organoids occurred in DO group, while more solid spherical organoids were seen in EO group, and the proportion of vacuoles was significantly greater in DO group than in EO group [(54.67±13.26)% vs (21.67±8.57)%, P < 0.05]. Short tandem repeat (STR) test of the nasal mucosal organoids and the source tissue showed a 100% match between them. On day 21 of culture, scanning and transmission electron microscopy of the nasal mucosal organoids identified ultrastructure of cilia in DO group and short villi structure in most of the organoids in EO group. Immunohistochemical staining showed positivity for P63 (basal cells), β-tubulin (ciliated columnar cells), and MUC5AC (goblet cells) in the organoids. Compared with those in EO group, the organoids in DO group showed significantly greater percentages of ciliated cells [(7.95±1.81)% vs (27.04±5.91)%, P < 0.05] and goblet cells [(14.46±0.93)% vs (39.85±5.43)%, P < 0.05) with a similar percentage of basal cells [(56.91±14.12)% vs (53.42±15.77)%, P > 0.05]. The differentiated nasal mucosal organoids in DO group were positively stained for glycogen.@*CONCLUSION@#The staged expansion-differentiation culture method allows more stable and prolonged growth of the cultured cells in vitro to produce organoids with controllable differentiation closely resembling the morphological structure and functions (ciliary function and secretory function) of the source tissue.
Subject(s)
Humans , Cell Differentiation , Cells, Cultured , Epithelial Cells , Nasal Mucosa , OrganoidsABSTRACT
Cancer is a threat to human health. New treatments for cancer can significantly prolong the survival time of patients， but fail to improve the adverse reactions induced by chemoradiotherapy. Improving patient outcomes still requires the effort of cancer researchers. In recent years， the anti-tumor effects of active components from Chinese herbs have received wide attention. Kaempferol， a flavonoid mainly found in the medicinal plant Kaempferia galanga， can be used to treat obesity， cardiovascular diseases， diabetes and other diseases. It has also exhibited good efficacy in inhibiting the occurrence and development of liver cancer， colon cancer， lung cancer， ovarian cancer， and other malignant tumors. Kaempferol mainly exerts the anti-cancer effect by inducing apoptosis. Specifically， it promotes the production of intracellular reactive oxygen species （ROS） and triggers cell apoptosis through the mitochondrial pathway. Besides， it is capable of interfering with the cancer cell cycle， causing most cancer cells to arrest in the G2/M phase， and inducing cell autophagy， a programmed cell death， thus inhibiting cell migration and invasion and angiogenesis， synergistically improving chemotherapeutic drug efficacy， and reducing adverse effects. Kaempferol acts on a series of intracellular and extracellular targets to participate in the regulation of tumor cell signaling pathways， involving phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）， epidermal growth factor receptor （EGFR）， mitogen activated protein kinase （MAPK）， and Wnt signaling pathways， with the PI3K/Akt signaling pathway being most significant. In addition， kaempferol also plays an important regulatory role in tumor epigenetics. This paper reviewed the anti-tumor effect and mechanism of kaempferol， aiming to provide reference for in-depth study on its prevention and treatment of tumor and the development of new anti-tumor drugs.
ABSTRACT
OBJECTIVE@#To establish a method of fingerprint position, sample transfer and fingerprint DNA extraction in contact samples.@*METHODS@#Sixty-six cases were visualized by 502 glue fingerprint fumigation. Two methods, ordinary wipe and acetone wipe, were used to transfer cast-off cells of fingerprints from testing samples, respectively. DNA was extracted and purified by ultramicro magnetic bead kit. The data was resolved on genetic analysis after amplification.@*RESULTS@#In 33 samples, 30 samples got better STR analysis by acetone wipe method. The peak range was 1,000-4,000 RFU and peak shapes were equable. It was hard to get ideal STR typing by ordinary wipe method.@*CONCLUSION@#The samples are visualized by 502 glue fingerprint fumigation and the case-off cells are transferred by acetone wipe method. The method shows better STR analysis result, which might be a better method for forensic science practice.