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OBJECTIVES@#To investigate the effect of recombinant human fibroblast growth factor 21 (rhFGF21) on the proliferation and mineralization of cementoblasts and its mechanism.@*METHODS@#Hematoxylin eosin, immunohistochemical staining, and immunofluorescence were used to detect the expression and distribution of fibroblast growth factor 21 (FGF21) in rat periodontal tissues and cementoblasts (OCCM-30), separately. Cell Counting Kit-8 was used to detect the proliferation of OCCM-30 under treatment with rhFGF21. Alkaline phosphatase staining and Alizarin Red staining were used to detect the mineralization state of OCCM-30 after 3 and 7 days of mineralization induction. The transcription and protein expression of the osteogenic-related genes Runx2 and Osterix were detected by real-time quantitative polymerase chain reaction (PCR) and Western blot analysis. The expression levels of genes of transforming growth factor β (TGFβ)/bone morphogenetic protein (BMP) signaling pathway in OCCM-30 were detected through PCR array analysis.@*RESULTS@#FGF21 was expressed in rat periodontal tissues and OCCM-30. Although rhFGF21 had no significant effect on the proliferation of OCCM-30, treatment with 50 ng/mL rhFGF21 could promote the mineralization of OCCM-30 cells after 7 days of mineralization induction. The transcriptional levels of Runx2 and Osterix increased significantly at 3 days of mineralization induction and decreased at 5 days of mineralization induction. Western blot analysis showed that the protein expression levels of Runx2 and Osterix increased during mineralization induction. rhFGF21 up-regulated Bmpr1b protein expression in cells.@*CONCLUSIONS@#rhFGF21 can promote the mineralization ability of OCCM-30. This effect is related to the activation of the TGFβ/BMP signaling pathway.
Subject(s)
Humans , Rats , Animals , Dental Cementum , Core Binding Factor Alpha 1 Subunit/metabolism , Cell Differentiation , Bone Morphogenetic Proteins/metabolism , Transforming Growth Factor beta/pharmacologyABSTRACT
OBJECTIVE：To improve the quality standard of Taraxaci Herba ，and to evaluate the quality of T. officinale from different origins. METHODS ：Based on the provisions of the 2020 edition of Chinese Pharmacopoeia （part Ⅰ）under the item “Taraxaci Herba ”，the method of content determination was added for the detection of water-soluble extracts （hot extraction method）and alcohol-soluble extracts （hot extraction method ），total flavonoids ，chlorogenic acid ，caffeic acid ，cichoric acid and isochlorogenic acid A. HPLC fingerprint was established by using 42 batches of T. officinale from 8 producing areas as object ，and principal component analysis was performed on the basis of above results. RESULTS ：The contents of alcohol-soluble extracts in 42 batches of T. officinale were 15.30％-30.40％，and those of water-soluble extracts were 27.59％-38.96％. The concentration of total flavonoids（UV spectrophotometry ），chlorogenic acid ，caffeic acid ，cichoric acid and isochlorogenic acid A （HPLC method ）were 0.016-0.096，0.003-0.196，0.004-0.117，0.025-1.578，0.002-0.152 mg/mL，respectively （all R2＞0.999）；RSDs of precision ， stability and repeatability tests were all lower than 2.00％（n＝6）；average sample recovery were 98.97％-103.53％，and RSDs were 1.19％-1.58％. The contents of total flavonoids ，chlorogenic acid ，caffeic acid ，cichoric acid and isochlorogenic acid A were 0.734％ -3.700％ ，0.004％ -0.123％ ，0.006％ - 0.087％ ，0.073％ -1.499％ ，0.005％ -0.109％ respectively in 42 batches of T. officinale. For 42 batches of T. officinale samples in HPLC fingerprint ，RSDs of the relative retention time of the common peak were 0-0.94％，and RSDs of the relative peak area were 0-125.57％. Among them ，the similarity of 39 batches of samples was all higher than 0.900. Results of principal component analysis showed that the quality of T. officinale from Shaanxi province was better，followed by medicinal materials from Hebei province. CONCLUSIONS ：Tentatively，the contents of alcohol-soluble extract，water-soluble extract ，total flavonoids ，chlorogenic acid ，caffeic acid ，cichoric acid and isochlorogenic acid A shall not be less than 17.0％，27.0％，1.383％，0.024％，0.021％，0.450％，0.021％ for Taraxaci Herba. In addition to the low content of caffeic acid in T. officinale from Shaanxi province ，the other indexes were better ；the content of caffeic acid in T. officinale from Hebei province was higher than that from Shaanxi province ，and other indicators were slightly lower than that from Shaanxi province. The quality of comprehensive evaluation of T. officinale from other origins was relatively poor ，and the quality of different batches of medicinal materials from the same origin was unstable.
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Neuroendocrine prostate cancer(NEPC) is a prostate cancer subtype with a very high degree of malignancy and a special molecular phenotype.NEPC is not sensitive to endocrine therapy, and there are currently no specific drugs, so there is a lack of effective clinical treatment.New advances in NEPC therapeutic include chemical therapy, targeted drug therapy based on molecular phenotype and other non-targeted drug therapy. This article summarizes the current treatment methods, pharmaceutical, and clinical research results for NEPC, aiming to deepen clinicians' more comprehensive understanding of NEPC patients' treatment strategies.
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Objective:To study the expression characteristics of Dickkopf1 (DKK1) in different time and space during tooth development of the postnatal mice, and to provide the theoretical basis for clarifying the mechanism of Wnt signaling pathway in regulating the tooth development.Methods:The postnatal Kunming mice at days 0.5, 6.5, 12.5, 18.5, 24.5, and 30.5 respectively after birth were selected and divided into various groups by time,three in each group.The mice in each group were sacrificed and the paraffin sections of mandibular bone including the first molar were prepared at the thickness of 5 μm, followed by HE staining and immunohistochemical staining in order to detect the expressions of DKK1 in tooth tissue and periodontal tissue.Results:At 0.5 d after birth, the mandibular first molar tooth germ was in the bell stage.At 6.5 d the enamel development of mandibular first molar was almost completed, and the epithelium root sheath extended to the root direction.At 12.5 d the dentin development of crown was completed, with the root formatted about 1/3. At 18.5 d the root had formatted about 2/3.At 24.5 d the root had reached the full length.At 30.5 d the apical foramen was narrow, and the root development was basically completed.There was no DKK1 expression at 0.5 d, but it expressed in the odontoblasts and predentin at 6.5 d. From days 12.5 to 30.5,the expressions of DKK1 were positive in periodontal ligament, alveolar bone, and cellular cementum as odontoblasts, which were gradually increased with the prolongation of time.However, no expression of DKK1 was detected in the pulp.Conclusion:DKK1 shows regular expressions at different tooth developmental stages after birth, suggesting its potential role in the growth of dentin and periodontal tissues.