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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
Objective: To assess the relative contributions of postharvest processing and geographical source to phytochemical variation of Corydalis Rhizoma, and rhizome of Corydalis yanhusuo, and to examine what phytochemical components are the most sensitive to the differences of each factor and how they change. Methods: HPLC fingerprinting and LC-MS coupled with chemometric approaches were applied. Results: The results of principal component analysis (PCA) and hierarchical cluster analysis (HCA) explicitly demonstrated the postharvest processing could produce a greater impact on the phytochemical profiles of Corydalis Rhizoma than geographical source. The contents of most compounds increased after water boiling while decreased after sulphur-fumigation. Protopine, coptisine, and palmatine were the most variable components in processing. Geographical sources also led to a remarkable phytochemical differentiation, in which the environmental variation of the three regions might play a role. Dehydrocorybulbine, coptisine, dehydrocorydaline, and protopine varied most among the three production regions and decreased sequentially in Zhejiang, Shaanxi, and Jiangsu provinces, China. Conclusion: Both postharvest processing and geographical source should be enhanced with the priority for the former in the quality control of Corydalis Rhizoma. The application of boiling is supported but the consistency should be improved in practice. Sulphur-fumigation is strongly suggested to be abandoned. © 2013 Tianjin Press of Chinese Herbal Medicines.
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With the completion of human genome, we are entering the functional genomic age. To further reveal the nature of life, glycomics comes into being and becomes a research focus. This review gives a deep insight on the current research progress of glycomics, including the research target, the structural classification and metabolism of sugar chain, the biological significance, research approaches, and the correlation between glycomics and liver diseases.
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Objective: To retrospectively analyze the routine clinical laboratory parameters for hepatocellular carcinoma, in an attempt to search for parameters for diagnosis of HCC. Methods: The pre-operation clinical laboratory data, such as tumor makers, and serological biochemical indices, hepatitis B virus (HBV) infection markers, and HBV DNA titers, were collected from 828 patients who were pathologically diagnosed as having HCC; then the correlation between these data with tumor size and the pathological grades of HCC was analyzed. Results: It was found that 97.9% of the 828 patients were infected with HBV and 70.9% of them were accompanied by liver fibrosis. We also found that the tumor size was correlated with albumin (Alb), globulin (GLB), A/ G, spartate aminotransferase (AST), ratio of aspartate to alanine aminotransferase (AST/ALT), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), alpha-L-fucosidase (AFU), alpha fetoprotein (AFP) and tumor grades; meanwhile, the pathological grades of tumor was correlated to PALB, GGT and tumor size(all P<0.05). Conclusion: The progress of HCC is correlated with several clinical laboratory biochemistry indices and virus indices. The combined application of these indices is worth studying for auxiliary diagnosis of HCC.
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Objective: To investigate the association between polymorphisms in α1 (I) collagen (COL1A1) gene promoter region and hepatic cirrhotic/fibrotic patients with hepatitis B infection. Methods: Three polymorphisms within COL1A1 promoter region were genotyped in 111 Chinese liver cirrhotic patients with chronic HBV infection and 95 healthy controls by direct DNA sequencing. Allelic and genotypic associations of these polymorphisms with liver cirrhosis were examined. The association between haplotype distribution, haplotype frequency and liver cirrhosis was analyzed by haplotype software. Results: No polymorphisms at position - 1662del/insT of COL1A1 gene were observed in subjects of both groups. There was no statistical difference in the genotype frequencies at position - 1997T>G or - 1363C>G in both groups. The frequency of haplotype - 1997T/-1363C in the diseased group was significantly higher than that in the normal control group (PG, - 1363C>G might be correlated with liver cirrhosis.
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Objective: To compare the expression profiles of serum glycomics between hepatocellular carcinoma(HCC) patients and normal adults, in an attempt to search for new biomarkers for HCC. Methods: Serum samples were obtained from HCC patients with hepatitis B virus(HBV) infection (n = 100) and normal adults(n = 130). Profiles of serum N-glycome were determined by capillary electrophoresis (CE) based on DNA sequencing equipment. Results: Each sample had a profile with 11 peaks; the peaks were adjusted and quantified. Peaks 1, 2, 9, and 11 in HCC group were higher than those in the control group; peaks 6, 8 and 10 were lower in HCC group than those in control group (P<0.05). The changes of peak values of several peaks were correlated with tumor markers such as alpha-fetoprotein(AFP), HBV DNA copies, albumin and prothrombin time. Conclusion: N-glycome profiles in serum are significantly different between HCC patients and normal adults, which may provide evidence for clinical diagnosis of HCC.
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<p><b>OBJECTIVE</b>To identify compounds that may be responsible for catnip response of Actinidia macrosperma, and compare chemical compositions in the wild and in vitro regenerated plants.</p><p><b>METHODS</b>GC-MS and relative retention indices with n-alkanes as reference points were used for compound identification, and component relative percentage was calculated based on GC peak areas without using correction factors.</p><p><b>RESULTS</b>There are 28 compounds (92.72%) and 15 compounds (93.88%) identified in the essential oils from the wild and regenerated plants, respectively. Dihydronepetalactone, iridomyrmecin, and dihydroactinidiolide, which are believed to be attractive to felines, are present in both wild and regenerated plants. Actinine was not detected, and beta-pheylethyl alcohol was only present in wild plant. In addition, short-chain enol derivatives, messengers in chemical communication, are commonly present in wild plant of A. macrosperma, but absent in regenerated one.</p><p><b>CONCLUSION</b>Dihydronepetalactone, iridomyrmecin, and dihydroactinidiolide are responsible for the catnip response of A. macrosperma.</p>
Subject(s)
Animals , Cats , Actinidia , Chemistry , China , Nepeta , Chemistry , Oils, VolatileABSTRACT
T polymorphism,was selected as putative promoter.The recombinant constructions containing-1328-+812 of TGF-?_1 gene and CAT reporter gene(phTGF2.14T,phTGF2.14C)were constructed and transfected into HepG2 cells with liposomal trans- fection method,then the transfected HepG2 cells were treated with IL-10(4 ng/ml),HGF(10 ng/ml)or IFN-?(20 ng/ml). Reporter gene activity was analyzed by ELISA.Results:Reporter gene activity in cells transfected with phTGF2.14C was sig- nificantly higher than those transfected with phTGF2.14T(P
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Objective:To analyze the relationship between hepatitis B virus(HBV)gene mutation at 1896 in precore region with genotype and replication of HBV and the liver function of patients.Methods:HBV precore 1896 site mutation,the genotype of HBV and serum content of HBV DNA were determined by PCR in 60 patients positive of HBV DNA.Chemiluminescence miacropaticle immunoassay(CMIA)was used for detection of serum HBeAg and HBeAb.Liver function parameters were ob- tained by routine biochemistry method.Results:The alanine aminotransferase(ALT)level in HBV with 1896 site mutation was significantly higher than that in the wildtype virus.Site mutation at 1896 had no correlation with HBeAg,HBV genotype and HBV DNA content.HBV DNA content in patient with genotype C was significantly higher than that with genotype B(P
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Objective:To retrospectively analyze the routine clinical laboratory parameters for hepatocellular carcinoma,in an attempt to search for parameters for diagnosis of hepatocellular carcinoma(HCC).Methods:The pre-operation clinical labo- ratory data,such as tumor makers,and serological biochemical indices,hepatitis B virus(HBV)infection markers,and HBV DNA titers,were collected from 828 patients who were pathologically diagnosed as having HCC;then the correlation between these data with tumor size and the pathological grades of HCC was analyzed.Results:It was found that 97.9% of the 828 pa- tients were infected with HBV and 70.9% of them were accompanied by liver fibrosis.We also found that the tumor size was correlated with albumin(ALB),globulin(GLB),A/G,aspartate aminotransferase(AST),ratio of aspartate to alanine amin- otransferase(AST/ALT),gamma-glutamyl transferase(GGT),alkaline phosphatase(ALP),alpha-L-fucosidase(AFU),al- pha-fetoprotein(AFP)and tumor grades;meanwhile,the pathological grades of tumor was correlated to prealbumin(PALB), GGT and tumor size(all P
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With the completion of human genome,we are entering the functional genomic age.To further reveal the nature of life,glycomics comes into being and becomes a research focus.This review gives a deep insight on the current research progress of glycomics,including the research target,the structural classification and metabolism of sugar chain,the biological significance,research approaches,and the correlation between glycomics and liver diseases.