Your browser doesn't support javascript.
Show: 20 | 50 | 100
Results 1 - 20 de 3.292
J. appl. oral sci ; 31: e20220313, 2023. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1421901


Abstract Bone morphogenetic protein 9 (BMP9) tends to be associated with various inflammatory responses of diseases, but its relationship with pulpitis remains unknown. Objective This study aimed to evaluate the effects and mechanisms of BMP9 in pulpitis. Methodology A rat model of pulpitis was used to evaluate the expression of BMP9, which was also analysed in Porphyromonas gingivalis lipopolysaccharide (Pg-LPS)-stimulated human dental pulp cells (hDPCs). The effects and mechanism of BMP9 on the regulation of inflammatory factors and matrix metalloproteinase-2 (MMP2) were evaluated using real-time quantitative PCR, western blotting, and immunocytofluorescence. Moreover, the migration ability of THP-1 monocyte-macrophages, treated with inflammatory supernate inhibited by BMP9, was previously tested by a transwell migration assay. Finally, a direct rat pulp capping model was used to evaluate in vivo the influence of the overexpression of BMP9 in pulpitis. Results The expression of BMP9 decreased after 24 h and increased after 3 and 7 d in rat pulpitis and inflammatory hDPCs. The overexpression of BMP9 inhibited the gene expression of inflammatory factors (IL-6, IL-8, and CCL2) and the secretion of IL-6 and MMP2 in Pg-LPS-stimulated hDPCs. The level of phosphorylated Smad1/5 was upregulated and the levels of phosphorylated ERK and JNK were downregulated. The inflammatory supernate of hDPCs inhibited by BMP9 reduced the migration of THP-1 cells. In rat pulp capping models, overexpressed BMP9 could partially restrain the development of dental pulp inflammation. Conclusion This is the first study to confirm that BMP9 is involved in the occurrence and development of pulpitis and can partially inhibit its severity in the early stage. These findings provided a theoretical reference for future studies on the mechanism of pulpitis and application of bioactive molecules in vital pulp therapy.

Article in Chinese | WPRIM | ID: wpr-953946


Colorectal cancer (CRC) is one of the refractory malignant tumors of the digestive system worldwide. With limitations, the available clinical therapies are usually suspended or show unsatisfactory effect. Therefore, it is urgent to find and develop new candidate drugs specifically targeting the cancer with ideal efficacy, low toxicity, and low cost, and the solutions can be found in traditional Chinese medicine (TCM) which has a long history. In TCM, sovereign, ministry, assistant, and guiding medicinals are selected based on the syndrome differentiation, and it has shown remarkable efficacy on CRC in recent years. In particular, Chinese medicinal compounds and monomers from Chinese medicinals which have been applied in clinical settings are advantageous in the treatment of CRCs, as they improve the quality of life, alleviate clinical symptoms and toxic and side effects of chemotherapy, and prolong the survival of patients. Therefore, we retrieved the English and Chinese articles with "CRC", "TCM", "compound" and "monomer" as keywords, and summarized the progress in the treatment of CRC with Chinese medicinal compounds and monomers from Chinese medicinals from four aspects of "replenishing Qi and invigorating spleen", "clearing heat and removing toxin", "nourishing liver and kidney", and "tonifying Qi and nourishing blood". However, Chinese medicine features multiple components, multiple targets, and multiple pathways, and in-depth research should be carried out on the application of Chinese medicinal compounds and monomers from Chinese medicinals in the treatment of CRC, in an attempt to minimize the pain and side effects and maximize the therapeutic effect. This study is expected to provide new insight into the treatment of CRC and a reference for further research on the efficacy and mechanism of Chinese medicine.

Biol. Res ; 562023.
Article in English | LILACS-Express | LILACS | ID: biblio-1429910


Background Knowledge about regulating transcription factors (TFs) for osteoblastogenesis from mesenchymal stem cells (MSCs) is limited. Therefore, we investigated the relationship between genomic regions subject to DNA-methylation changes during osteoblastogenesis and the TFs known to directly interact with these regulatory regions. Results The genome-wide DNA-methylation signature of MSCs differentiated to osteoblasts and adipocytes was determined using the Illumina HumanMethylation450 BeadChip array. During adipogenesis no CpGs passed our test for significant methylation changes. Oppositely, during osteoblastogenesis we identified 2462 differently significantly methylated CpGs (adj. p < 0.05). These resided outside of CpGs islands and were significantly enriched in enhancer regions. We confirmed the correlation between DNA-methylation and gene expression. Accordingly, we developed a bioinformatic tool to analyse differentially methylated regions and the TFs interacting with them. By overlaying our osteoblastogenesis differentially methylated regions with ENCODE TF ChIP-seq data we obtained a set of candidate TFs associated to DNA-methylation changes. Among them, ZEB1 TF was highly related with DNA-methylation. Using RNA interference, we confirmed that ZEB1, and ZEB2, played a key role in adipogenesis and osteoblastogenesis processes. For clinical relevance, ZEB1 mRNA expression in human bone samples was evaluated. This expression positively correlated with weight, body mass index, and PPARγ expression. Conclusions In this work we describe an osteoblastogenesis-associated DNA-methylation profile and, using these data, validate a novel computational tool to identify key TFs associated to age-related disease processes. By means of this tool we identified and confirmed ZEB TFs as mediators involved in the MSCs differentiation to osteoblasts and adipocytes, and obesity-related bone adiposity.

Braz. j. biol ; 83: e246592, 2023. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1339408


Abstract Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37oC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.

Resumo As células-tronco mesenquimais (MSCs) possuem grande potencial para aplicação em procedimentos terapêuticos ligados a terapia celular e engenharia de tecidos, considerando-se a plasticidade e capacidade de formação em diferentes tipos celulares por elas. Dada a abrangência no emprego das MSCs, há necessidade de se compreender melhor algumas propriedades relacionadas à diferenciação osteogênica, particularmente liga à biomateriais usados em engenharia de tecidos. Este projeto objetiva o desenvolvimento de uma metodologia de análise empregando-se a FT-Raman para identificação e quantificação de componentes bioquímicos presentes em meios de cultura condicionados por MSCs, com ou sem indução à diferenciação osteogênica. Todos os experimentos foram realizados entre as passagens 3 e 5. Para essas análises, as MSCs foram cultivadas sobre arcabouços de polímeros biorreabsorvíveis de poli (hidroxibutirato-co-hidroxivalerato) (PHBV) e o poli (ε-caprolactona) (PCL). As MSCs (GIBCO®) foram inoculadas nos polímeros puros e na mistura 75:25 de PHBV / PCL (amostras densas e porosas). As células foram mantidas em DMEM (com baixa glicose) contendo GlutaMAX® e 10% de SFB a 37oC com 5% de CO2 por 21 dias. A própria placa foi usada como controle. Os meios de cultura condicionados foram coletados e analisadas em FT-Raman para sondagem de grupos funcionais, bem como possíveis variações moleculares associadas com a diferenciação e metabolismo celular. Foi possível discernir grupos funcionais de moléculas específicas no meio condicionado, como colesterol, fosfatidilinositol, triglicerídeos, forma Beta de polipeptídeos, regiões de amida e ligações de hidrogênio de proteínas, além da expressão de DNA. Na presente avaliação, a FT-Raman apresentou como uma técnica de resolução limitada, uma vez que modos vibracionais de estiramento próximos ou mesmo iguais podem ser expressos por moléculas diferente, dificultando a análise. Não houve variações nas leituras entre as amostras estudadas, concluindo-se que a FT-Raman não atendeu às expectativas nas condições estudadas.

Animals , Rats , Mesenchymal Stem Cells , Osteogenesis , Polyesters , Spectrum Analysis, Raman , Culture Media, Conditioned , Cell Proliferation , Tissue Scaffolds
Article in Chinese | WPRIM | ID: wpr-969308


@#Osteoclasts are the only cells responsible for bone resorption in the body, and osteoblasts are the main cells responsible for bone regeneration in the body. Under physiological conditions, these cells maintain a dynamic balance to maintain bone homeostasis. It was widely believed that the imbalance of bone metabolism is mainly affected by the expression of related inflammatory factors. However, with the gradual expansion of related studies in recent years, autophagy has been shown to be closely related to the differentiation, apoptosis and functions of osteoclasts and osteoblasts. AMP-activated protein kinase (AMPK) is an important regulator of energy metabolism in vivo and is involved in the regulation of autophagy and bone homeostasis in bone metabolism-related cells. Periodontitis is a chronic infectious disease, and its typical symptoms are alveolar bone resorption. At present, controlling the level of periodontal inflammation and alveolar bone resorption more effectively in clinical practice remains a challenge. The detection of AMPK and autophagy levels in bone metabolism-related cells shows certain prospects for the clinical prevention and treatment of periodontitis in the future. Therefore, this article reviews the regulation of periodontal inflammation levels and bone homeostasis through cell autophagy related to AMPK-mediated bone metabolism.

Article in Chinese | WPRIM | ID: wpr-965663


ObjectiveTo investigate the clinical value of Longbeisan application at Shenque (CV 8) combined with oral administration of Chinese medicine in the treatment of premature ejaculation (PE). MethodA total of 98 PE patients treated in the andrology department of the First Affiliated Hospital of Henan University of Chinese Medicine at the same time period were randomly assigned into an observation group and a control group, with 49 patients in each group. The observation group received Longbeisan application at Shenque (CV 8) combined with oral treatment of Chinese medicine according to syndrome differentiation, and the control group was treated with dapoxetine hydrochloride tablets. The treatment in both groups lasted for 8 weeks. The intravaginal ejaculatory latency time (IELT), Chinese index of premature ejaculation-5 (CIPE-5) score, patient's sexual life satisfaction, spouse's sexual life satisfaction, effective rate, and adverse reaction incidence were compared between the two groups. ResultAfter treatment, the observation group had higher total effective rate than the control group [(85.71% (42/49) vs. 67.35% (33/49), χ2=6.262, P<0.05]. The IELT, CIPE-5 score, and patient's and spouse's satisfaction scores after treatment increased compared with those before treatment (P<0.01), and the increases were more significant in the observation group (P<0.05, P<0.01). The clinical effect of the observation group was better than that of the control group. During the treatment, 7 (7/49,14.29%) patients in the control group and 2 ,2/49,4.08%) patients in the observation group showed adverse reactions, which indicated the safety of the observation group was better than that of the control group (χ2=9.000, P<0.05). In the follow-up period, 11 (11/49,22.45%) patients in the control group and 3 (3/49,6.12%) patients in the observation group showed aggravation of symptoms, which meant that the observation group had better lasting effect (χ2=0.317, P<0.05). ConclusionLongbeisan application at Shenque (CV 8) combined with oral administration of Chinese medicine has better clinical effect, stronger safety, and longer effect than dapoxetine hydrochloride in the treatment of PE.

Article in Chinese | WPRIM | ID: wpr-965642


Da Qinjiaotang is a common classical prescription for the treatment of stroke. It originates from Collection of Writings on the Mechanism of Disease, Suitability of Qi, and the Safeguarding of Life as Discussed in the Basic Questions (《素问病机气宜保命集》) by physician LIU Wansu, and is composed of Gentianae Macrophyllae Radix, Glycyrrhizae Radix et Rhizoma, Chuanxiong Rhizoma, Angelicae Sinensis Radix, Paeoniae Radix Alba, Asari Radix et Rhizoma, Notopterygii Rhizoma et Radix, Saposhnikoviae Radix, Scutellariae Radix, Gypsum Fibrosum, Angelicae Dahuricae Radix, Atractylodis Macrocephalae Rhizoma, Rehmanniae Radix, Rehmanniae Radix Praeparata, Poria, and Angelicae Pubescentis Radix. Doctors of all dynasties have disputed the composition principle of the prescription and argued whether its treatment of stroke belongs to the theory of "internal wind" or "external wind". Through collating and analyzing ancient and modern literature related to the indications of Da Qinjiaotang, this paper was dedicated to the origin of syndrome differentiation and treatment of Da Qinjiaotang. According to LIU Wansu's original works, Da Qinjiaotang is a prescription for the treatment of "internal wind", and in the prescription, wind medicinal herbs such as Gentianae Macrophyllae Radix, Notopterygii Rhizoma et Radix and Angelicae Pubescentis Radix removes stagnation, clears sweat pore, and makes qi and blood channels flow smoothly. However, later generations, affected by the idea of "external wind", believe that this prescription is used for the treatment of "external wind". Ancient physicians gradually supplemented the symptoms of stroke, such as wry eye and mouth, hemibody pain and limb numbness, which were treated by Da Qinjiaotang, and Da Qinjiaotang was also applied to the treatment of other diseases, such as tendon dryness, convulsion and arthralgia. Modern doctors still explain the disease pathogenesis from the theory of "external wind" as deficiency in channels and collaterals and the entry of pathogenic wind, and the prescription has the effect of dispersing wind, clearing heat and nourishing and activating blood. In clinical practice, Da Qinjiaotang is mainly used to treat cerebrovascular diseases and peripheral facial paralysis in nervous system diseases, gouty arthritis and rheumatic arthritis in the rheumatic immune system and skin diseases. The above findings facilitate the research and development of Da Qinjiaotang.

Article in Chinese | WPRIM | ID: wpr-965524


Objective To investigate the genetic diversity and genetic differentiation of different geographical isolates of Gohieria fusca.. Methods G. fusca isolates were sampled from Wuhu (WH), Bengbu (BB) and Bozhou cities (BZ) of Anhui Province and Jiaxing City of Zhejiang Province (JX). Mitochondrial cytochrome b (Cytb) and ribosomal internal transcribed spacer (ITS) genes were amplified in WH, BB, BZ and JX isolates of G. fusca using PCR assay. The gene sequences were edited and aligned using the software Chromas 2 and DNASTAR 1.00, and the haplotype, haplotype diversity (Hd) and nucleotide polymorphism (Pi) of each isolate were calculated using the software DnaSP 5.10.00. The genetic differentiation among isolates (Fst) and gene flow value (Nm) were estimated using the software MEGA 10.2, and a phylogenetic tree was built. Tests of neutrality and analysis of molecular variance (AMOVA) were performed using the software Arlequin 3.1 and a haplotype network was built based on the Median-Joining network using the software Network 10.2. Results PCR assay showed that the sizes of the Cytb and ITS genes were 372 bp and 1 301 to 1 320 bp, respectively. All four isolates of G. fusca presented high genetic diversity based on mitochondrial Cytb and ITS genes (Hd = 0.804, Pi = 0.006 91). AMOVA showed genetic differentiation among geographical isolates of G. fusca (Fst = 0.202 40, P < 0.05), and the genetic variation was mainly caused by intra-population variations (79.76%). Gene flow analysis showed a high level of gene flow among G. fusca isolates (Nm > 1). Tests of neutrality based on Cytb gene measured a Tajima’s D value of −1.796 31 (P < 0.05) and a Fu’s FS value of −3.293 98 (P < 0.05) in WH isolate of G. fusca, indicating population expansion in WH isolate of G. fusca. Haplotype network analysis and phylogenetic analysis revealed no remarkable geographical distribution pattern among different geographical isolates of G. fusca. All four isolates of G. fusca presented high genetic diversity (Hd = 0.985, Pi = 0.011 97). AMOVA showed moderate level of genetic differentiation between four isolates (Fst = 0.104 62, P < 0.05). The tests of neutrality based on ITS genes measured a Tajima’s D value of −6.088 20 and a Fu’s FS value of −1.935 99 (both P > 0.05) in the whole isolate of G. fusca, indicating no obviously population expansion. Conclusions The four geographical isolates of G. fusca have high genetic diversity and remarkable genetic differentiation. Since a high level of gene flow is detected among different geographical isolates of G. fusca, no obvious geographical distribution pattern of G. fusca is found.

Article in Chinese | WPRIM | ID: wpr-964433


Objective@# To explore the effects of red LED light mediated by the Kelch-like ECH-associated protein 1-nuclear factor erythroid 2-related factor 2/heme oxygenase-1 (KEAP1-NRF2/HO-1) pathway on osteogenic differentiation and oxidative stress damage of human periodontal ligament stem cells (hPDLSCs) induced by high glucose, which provides a basis for the application of red light-emitting diode (LED) light in cell antioxidative damage.@*Methods@#hPDLSCs were identified by flow cytometric analysis, alkaline phosphatase (ALP) staining and Alizarin red-S staining; hPDLSCs were pretreated in a high glucose environment for 48 hours and irradiated with 1, 3, or 5 J/cm2 red LED light. A CCK-8 assay was performed to choose the radiant exposure that had the strongest effect on promoting the cell proliferation rate for subsequent experiments. hPDLSCs were divided into a control group, a high glucose group and a high glucose+light exposure group. ALP staining, ALP activity, Alizarin red-S staining and quantitative calcified nodules were used to detect the osteogenic differentiation of hPDLSCs; qRT-PCR and Western blot were used to detect the gene and protein expression levels of ALP, runt-related transcription factor 2 (RUNX2) and osterix (OSX); the relative mRNA expression levels of antioxidant enzyme-related genes superoxide dismutase 2 (SOD2) and catalase (CAT) in hPDLSCs were detected by qRT-PCR; reactive oxygen species (ROS) levels were detected by fluorescence microscopy and flow cytometry; the tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels in cell supernatants were detected by ELISA; the NRF2-specific inhibitor ML385 was used to inhibit the NRF2 pathway; ALP staining and ALP activity were used to detect the markers of early osteogenic differentiation; qRT-PCR was used to detect the gene expression of ALP, RUNX2 and OSX; and the protein expression levels of KEAP1, NRF2 and HO-1 were detected by Western blot.@*Results @# Identified, and irradiant exposure of 5 J/cm2 was chosen for subsequent experiments. Red LED light irradiation (5 J/cm2) improved the osteogenic differentiation of hPDLSCs induced by high glucose (P<0.05), increased the mRNA and protein levels of ALP, RUNX2 and OSX (P<0.05), upregulated the mRNA expression levels of SOD2 and CAT (P<0.05), reduced the levels of ROS (P<0.05), and reduced TNF-α and IL-1β levels in the cell supernatants (P<0.05). When ML385 was added to inhibit the NRF2 pathway, the ALP activity of cells was decreased (P<0.05); the gene expression levels of ALP, RUNX2 and OSX were downregulated (P<0.05); the protein level of KEAP1 was upregulated (P<0.05); and the protein levels of NRF2 and HO-1 were downregulated (P<0.05)@*Conclusion@#Red LED light may promote the proliferation and osteoblastic differentiation of hPDLSCs induced by high glucose through the KEAP1-NRF2/HO-1 pathway and reduce the oxidative stress damage to hPDLSCs induced by high glucose.

Article in Chinese | WPRIM | ID: wpr-964281


Objective To explore the effects of Cordyceps sinensis extract (CSE) on osteoporosis and RANKL-mediated osteoclastogenesis. Methods Bone marrow-derived macrophages (BMMs) was isolated from the bone marrow of C57BL/6 mice. CSE was added in osteoclast differentiation. Osteoclasts were stained by tartrate-resistant acid phosphatase (TRAP). The nearly mature osteoclasts were planted on hydroxyapatite plates and the area of bone lacunae was observed by microscope. The F-actin belt was stained by DAPI and phylloeptide and the number of nuclei was observed by confocal microscopy. The expressions of DC-STAMP, ATP6V0D2, TRAP, CTSK, and NFATC1 were detected by q-PCR. The protein expression of the MAPK pathway was detected by Western Blot. The in vivo experiments were carried out by administering CSE to the ovariectomized mice daily through gavage. After 6 weeks of intervention, mouse femurs were taken for morphological analysis. Peripheral blood was taken for ELISA. Results CSE represses osteoclastogenesis, bone resorption, F-actin belts formation, osteoclast specific gene expressions and MAPK signaling pathways in vitro. In vivo study indicated that CSE prevents OVX-induced osteoporosis and preserves bone volume by repressing osteoclast activity and function. It also increases the serum ALP, BGP content, and reduces TRAP content. Conclusion CSE can attenuate osteoclast formation and OVX-induced osteoporosis, suggesting potential clinical therapeutic effects for osteoporosis.

Article in Chinese | WPRIM | ID: wpr-961149


Objective @#To investigate the role and mechanism of bone formation caused by the ratio of advanced platelet-rich fibrin (A-PRF) and β-tricalcium phosphate (β-TCP) in rabbit femur defect model, which provides a new idea for clinical treatment of bone defect.@*Methods @#Twenty-four New Zealand white rabbits were divided into model group, 1∶1 complex group (A-PRF∶β-TCP=1∶1), 2∶1 complex group (A-PRF∶β- TCP=2∶1) and 4∶1 complex group (A-PRF∶β- TCP=4∶1), with 6 rabbits in each group. Femoral defect models were constructed in each group. In the composite group, the bone defect was filled with composite material, while in the model group, no material was filled. After 8 weeks, the animals were euthanized and specimens were collected. Bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular separation (Tb.SP) and trabecular number (Tb.N) in femoral defect tissue were measured by micro-CT and photographed. Hematoxylin - eosin staining was used to detect the pathological changes of new bone tissue. The morphological changes of the new bone tissue were observed by scanning electron microscopy. Determination of phospho-mitogen activated protein kinase p38 (p-p38MAPK), CCAAT/enhancer binding protein homologous protein (CHOP) and phospho-cysteine aspartic protease-3 (p-Caspase3) in newborn femur by ELISA. The mRNA expressions of osteoprotegerin (OPG), bone morphogenetic protein-2 (BMP-2), receptor activator of nuclear factor kappa-B ligand (RANKL) and p38MAPK were detected by real-time quantitative PCR. The expression of OPG, BMP-2, RANKL, p-p38MAPK and p-Caspase3 protein in the new bone tissue was observed by immunohistochemistry. @*Results @#In the model group, bone formation in the femoral defect area was slow and osteogenic quality was poor. Compared with the model group, the bone formation and neocapillaries of femoral defect area in the complex group was good, BMD, BV.TV, Tb.Th, Tb.N were increased, and Tb.Sp were decreased, the expressions of p-p38MAPK, CHOP and p-Caspase3 were decreased, and the mRNA and protein expressions of OPG and BMP-2 were increased. The mRNA expression of RANKL and p38MAPK was decreased. Apoptosis in new bone tissue of each group showed the lowest apoptosis rate in samples of the 2∶1 complex group (P<0.05); A-PRF: β-TCP=2∶1 ratio has the best osteogenic effect. @*Conclusion@#The complex composed of A-PRF and β-TCP can promote the expression of OPG, inhibit the expression of RANKL and phosphorylation of p38MAPK, reduce the apoptosis of new bone tissue cells, and promote osteogenic differentiation.

Article in Chinese | WPRIM | ID: wpr-960928


Respiratory diseases are common, frequently-occurring clinical diseases. As the prevalence rate is increasing year by year, they have become a problem that seriously affects public health. The diseases are mainly located in the lung by traditional Chinese medicine (TCM) syndrome differentiation. Lung governs Qi and controls breathing and is also an organ for the storage of phlegm. Clinically, phlegm and Qi are often used for the treatment. Banxia Houputang (BHT), originated from Synopsis of the Golden Chamber (《金匮要略》), was used to treat plum-stone Ai (globus hystericus) at first. It is composed of Rhizoma Pinelliae, Cortex Magnoliae Offcinalis, Poria, Rhizoma Zingiberis Recens, and Folium Perillae, and treats diseases with the core pathogensis of mutual obstruction of phlegm and Qi. BHT has the effects of moving Qi, dissipating mass, descending adverse Qi, and resolving phlegm, which basically correspond to the pathological characteristics of the lungs. Clinical studies have confirmed that modified BHT can be used either alone or in combination with western medicine to treat chronic pharyngitis, asthma, chronic obstructive pulmonary disease, pneumonia, obstructive sleep apnea, upper airway cough syndrome and other respiratory diseases, with significant effects. It effectively improves the symptoms and signs of the diseases and reduces the recurrence rate. Basic research has shown that BHT plays anti-inflammatory, anti-oxidative stress, anti-apoptotic, autophagy-regulating, and iron overload-regulating roles by regulating the targets in multiple pathways. This paper, by combing the relevant literature in recent years, conducted a systematic review on BHT from the three aspects of syndrome analysis, clinical treatment research and mechanism research, with a view to providing theoretical basis and reference for the mechanism research of BHT in treating respiratory diseases and for expanding its clinical application.

Ter. psicol ; 40(3)dic. 2022.
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1424678


Antecedentes La diferenciación del self es la capacidad intrapsíquica de distinguir las emociones de las cogniciones, y la capacidad interpersonal de mantener vínculos significativos y ser autónomos. Objetivo Analizar la relación entre la diferenciación y las habilidades sociales, y las diferencias en la diferenciación en función de la asistencia a psicoterapia en una muestra española. Método La muestra está formada por 126 sujetos españoles, 78 mujeres y 48 hombres, de entre 18 y 65 años, que contestaron un cuestionario sociodemográfico, la Escala de Diferenciación del Self y la Escala de Habilidades Sociales. Resultados Se observaron relaciones entre la diferenciación y las habilidades sociales. Además, la fusión con los otros y el corte emocional predecían las habilidades sociales. Por otro lado, los sujetos que habían realizado una terapia anteriormente y aquellos que nunca habían acudido a terapia tenían unos niveles de diferenciación más altos que aquellos que acudían a terapia en el momento del estudio. Conclusiones Existen asociaciones entre la diferenciación del self, las habilidades sociales y la asistencia a terapia. Se discuten los resultados y se sugieren futuras líneas de investigación.

Background Differentiation of self is the intrapsychic capacity to distinguish emotions from cognitions and the interpersonal capacity to maintain significant bonds and to be autonomous. Objetive To analyze the relationship between differentiation and social skills, and the differences in differentiation according to therapy attendance in a Spanish sample. Method The sample consisted of 126 Spanish subjects, 78 women and 48 men, aged between 18 and 65 years, who answered a sociodemographic questionnaire, the Differentiation of Self Scale and the Social Skills Scale. Results Significant relationships between differentiation of self and social skills were observed. Furthermore, fusion with others and emotional cutoff predicted social skills. On the other hand, subjects who had previously undergone therapy and those who had never attended therapy had higher levels of differentiation than those who were attending therapy at the time of the study. Conclusion There are associations between differentiation of self, social skills, and therapy attendance. Results are discussed and future lines of research are suggested.

Braz. dent. j ; 33(5): 35-45, Sep.-Oct. 2022. graf
Article in English | LILACS-Express | LILACS, BBO | ID: biblio-1403781


Abstract To investigate osteoclast formation in vivo and if leukotriene B4 (LTB4) loaded in microspheres (MS) could be used as a therapeutical strategy to promote a sustained delivery of the mediator and prevent osteoclast differentiation. Methods: In vivo, apical periodontitis was induced in mice to investigate osteoclast differentiation and signaling in absence of 5-lipoxygenase (5-LO). In vitro, LTB4-MS were prepared using an oil-in-water emulsion solvent extraction-evaporation process. Characterization and efficiency of LTB4 encapsulation were investigated. J774A.1 macrophages were cultured in the presence of monocyte colony-stimulating factor (M-CSF) and ligand for receptor activator of nuclear factor kappa B (RANKL) and then stimulated with LTB4-MS. Cytotoxicity, in vitro MS-LTB4 uptake, osteoclast formation and gene expression were measured. Results: We found that 5-LO negatively regulates osteoclastic formation in vivo during apical periodontitis development. In vitro, LTB4-MS were up-taken by macrophages and were not cytotoxic to the cells. LTB4-MS inhibited osteoclast formation and the synthesis of osteoclastogenic genes Acp5, Mmp9, Calcr and Ctsk. LTB4-MS inhibited differentiation of macrophages into an osteoclastic phenotype and cell activation under M-CSF and RANKL stimulus.

Resumo O objetivo deste trabalho foi Investigar a formação de osteoclastos in vivo e se o leucotrieno B4 (LTB4) incorporado em microesferas (MS) poderia ser usado como estratégia terapêutica para promover uma entrega sustentada do mediador e prevenir a diferenciação dos osteoclastos. Métodos: In vivo, a periodontite apical foi induzida em camundongos para investigar a diferenciação e sinalização de osteoclastos na ausência de 5-lipoxigenase (5-LO). In vitro, LTB4-MS foi preparado usando um processo de evaporação e extração de solvente de emulsão de óleo em água. A caracterização e a eficiência do encapsulamento do LTB4 foram investigadas. Macrófagos J774A.1 foram cultivados na presença de fator estimulador de colônia de monócitos (M-CSF) e ligante para o receptor ativador do fator nuclear kappa B (RANKL) e, então, estimulados com LTB4-MS. Citotoxicidade, captação in vitro de MS-LTB4, formação de osteoclastos e expressão gênica foram avaliadas. Resultados: A via 5-LO regula negativamente a formação de osteoclastos in vivo durante o desenvolvimento da periodontite apical. In vitro, LTB4-MS foram fagocitadas pelos macrófagos e não foram citotóxicos para as células. LTB4-MS inibiu a formação de osteoclastos e a síntese dos genes pró-osteoclastogênicos Acp5, Mmp9, Calcr e Ctsk. Conclusões: LTB4-MS inibiu a diferenciação de macrófagos em um fenótipo osteoclástico e a ativação celular sob estímulo de M-CSF e RANKL.

Rev. latinoam. psicopatol. fundam ; 25(3): 643-665, 2022.
Article in Portuguese | LILACS-Express | LILACS, INDEXPSI | ID: biblio-1424070


O objetivo do presente artigo é analisar a estreita relação entre o fenômeno das automutilações e a problemática dos limites, tendo como ponto de partida o ataque à pele como condição de possibilidade para essa prática. A pele remete à sensorialidade e às primeiras noções de eu, tendo como função proteger nossa individualidade e fundamentar nossa troca com o outro. Ela contém uma premissa de integração, constituindo-se como uma fronteira que marca os limites da própria existência. Nas automutilações, a agressão à pele denuncia um prejuízo na construção da diferenciação entre sujeito e objeto, deixando em evidência uma confusão nos limites do sujeito. Nossa hipótese é a de que o recurso ao corpo, mais especificamente o ataque à pele, nas automutilações, surge como uma tentativa de contenção do eu em momentos nos quais o sujeito sente que pode haver o risco da perda da integridade narcísica.

Resumos This paper analyzes the close relationship between self-mutilation and the problem of limits, understanding the attack on the skin as a condition of possibility for this practice. The skin, which leads us to sensoriality and the first notions of self, acts to protect our individuality and support our interactions with the Other. It contains a premise of integration, constituting a boundary that marks the limits of existence itself. In self-mutilations, the attack on the skin reveals a loss in the the differentiation between subject and object, revealing a confusion in the subject's limits. The text posits that the recourse of the body, by attacking the skin, in self-mutilations, emerges as an attempt to contain the self in moments when the subject feels at risk of losing their narcissistic integrity.

Cet article analyse la relation étroite entre le phénomène d'automutilation et le problème des limites, en comprenant l'attaque de la peau comme une condition de possibilité de cette pratique. La peau, qui renvoie à la sensorialité et aux premières notions de soi, agit pour protéger notre individualité et soutenir nos interactions avec l'Autre. Elle contient une prémisse d'intégration, constituant une frontière qui marque les limites l'existence elle-même. Dans les automutilations, l'atteinte à la peau révèle une perte dans la construction de la différenciation entre sujet et objet, témoignant d'une confusion dans les limites du sujet. Le texte postule que le recours au corps, en attaquant la peau, dans les automutilations, émerge comme une tentative de contenir le soi dans les moments où le sujet sent en danger de perdre son intégrité narcissique.

El objetivo de este artículo es analizar la estrecha relación entre el fenómeno de las automutilaciones y el problema de los límites, teniendo como punto de partida el ataque a la piel como condición de posibilidad para esta práctica. La piel se refiere a la sensorialidad y a las primeras nociones del yo, teniendo como función proteger nuestra individualidad y fundamentar nuestro intercambio con el otro. Contiene una premisa de integración, constituyendo una frontera que marca los límites de la existencia misma. En las automutilaciones, la agresión a la piel revela una pérdida en la construcción de la diferenciación entre sujeto y objeto, evidenciando una confusión en los límites del sujeto. Nuestra hipótesis es que recurrir al cuerpo, más específicamente atacar la piel, en las automutilaciones, aparece como un intento de contener al yo en momentos en los que el sujeto siente que puede haber riesgo de pérdida de la integridad narcisista.

Rev. peru. med. exp. salud publica ; 39(2): 227-235, abr.-jun. 2022. tab, graf
Article in Spanish | LILACS | ID: biblio-1395048


RESUMEN Los organoides son estructuras miniaturizadas, generadas principalmente a partir de células madre pluripotentes inducidas, que se cultivan en el laboratorio conservando sus características innatas o adquiridas. Tienen el potencial de reproducir procesos de desarrollo biológico, modelar procesos patológicos que permitirán el descubrimiento de nuevos fármacos y propicien la medicina regenerativa. Sin embargo, estas experiencias requieren perfeccionamiento constante porque pueden haberse realizado variaciones en la constitución de estos órganos. Por ello, el presente artículo tiene como objetivo revisar la información actualizada sobre organoides y sus procesos experimentales básicos y recientes, empezando por la gastrulación, para tratar de imitar, en lo posible, la formación de las tres capas: ectodermo, mesodermo y endodermo, incluyendo los factores que intervienen en la inducción, diferenciación y maduración en la generación de estos organoides. Asimismo, el diseño y preparación de medios de cultivo altamente especializados que permitan obtener el órgano seleccionado con la mayor precisión y seguridad. Se realizó una búsqueda de artículos originales y de revisión publicados en PubMed, Nature y Science. Los artículos se seleccionaron por sus resúmenes y por su texto completo. Las conclusiones de este articulo destacan las ventajas futuras en el uso y aplicaciones de los organoides.

ABSTRACT Organoids are tiny structures, mainly generated from induced pluripotent stem cells, which are cultured in the laboratory while retaining their innate or acquired characteristics. They have the potential to reproduce biological development processes, model pathological processes that will enable the discovery of new drugs and promote regenerative medicine. However, these processes require constant improvement because variations may have occurred in the constitution of the organs. Therefore, this article aims to review updated information on organoids and their basic and recent experimental processes, starting with gastrulation, in an attempt to mimic, as much as possible, the formation of the three layers: ectoderm, mesoderm and endoderm; as well as the information regarding the factors involved in the induction, differentiation and maturation during the generation of organoids. Likewise, the design and preparation of highly specialized culture media that allow obtaining the selected organ with the highest precision and safety. We searched for original and review articles published in PubMed, Nature and Science. Articles were selected for their abstracts and full text. The conclusions of this article highlight the future advantages in the use and applications of organoids.

Organoids , Signal Transduction , Cell Differentiation , Gastrulation , Induced Pluripotent Stem Cells
Clinics ; 77: 100019, 2022. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1375188


Abstract Objectives Osteoblasts are derived from Bone Marrow-derived Mesenchymal Stem Cells (BM-MSCs), which play an indispensable role in bone formation. In this study, the authors aim to investigate the role of IRF4 in the osteogenic differentiation of BM-MSCs and its potential molecular mechanism. Methods The authors used lentivirus infection to overexpress IRF4 in BM-MSCs. The expression of IRF4 and osteogenesis-related genes were detected by qRT-PCR and western blot analysis. The osteogenic differentiation of BM-MSCs was evaluated by Alkaline Phosphatase (ALP) activity, Alizarin red staining, and Alkaline Phosphatase (ALP) staining. Chromatin Immunoprecipitation (ChIP), Dual-Luciferase reporter assay and RNA Immunoprecipitation Assay were applied to confirm the regulatory mechanism between IRF4, miR-636 and DOCK9. Results The authors found IRF4 was down-regulated during the osteogenic differentiation of BM-MSCs, and IRF4 overexpression could decrease the osteogenic differentiation of BM-MSCs by specifically promoting the reduction of Alkaline Phosphatase (ALP) activity and down-regulating osteogenic indicators, including OCN, OPN, Runx2 and CollA1. Mechanistically, IRF4 activated microRNA-636 (miR-636) expression via binding to its promoter region, and Dedicator of Cytokinesis 9 (DOCK9) was identified as the target of miR-636 in BM-MSCs. Moreover, the damage in the capacity of osteogenic differentiation of BM-MSCs induced by IRF4 overexpression could be rescued by miR-636 inhibition. Conclusions In summary, this paper proposed that IRF4/miR-636/DOCK9 may be considered as targets for the treatment of osteoporosis (OP).

Article in Chinese | WPRIM | ID: wpr-956550


Objective:To prepare the hydrogel scaffolds with different concentrations of laponite and compare their osteogenic properties.Methods:The scaffolds of gelatin/sodium alginate hydrogel into which laponite was added according to the mass ratios of 0%, 1%, 2%, and 3% were assigned into groups T0, T1, T2, and T3. In each group, the compressive modulus was measured and the leaching solution for 24 h extracted to measure the ion release. Bone marrow mesenchymal stem cells (BMSCs) were cultured in the extract medium from each group and common medium (blank group) ( n=3) in the in vitro experiments to determine the expression of osteogenic genes Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and type I collagen after 7 days of culture. In the in vivo experiments, the scaffolds were implanted into the femoral condyle defects in rats, and a blank group with no scaffolds was set. The bone repair in each group was evaluated by hematoxylin-eosin(HE) staining and immunohistochemical staining. Results:The compressive modulus in group T2 [(139.05±6.43) kPa] was significantly higher than that in groups T0, T1 and T3 [(68.83±3.76) kPa, (101.18±3.68) kPa and (125.40±3.28) kPa] ( P<0.05). The ion contents of lithium, magnesium and silicon released from the 24 h leaching solution in group T2 were (0.031±0.005) μg/mL, (3.047±0.551) μg/mL and (5.243±0.785) μg/mL, insignificantly different from those in group T3 ( P> 0.05) but significantly larger than those in group T1 ( P>0.05). The in vitro experiments showed that the expression levels of Runx2, ALP and type I collagen in group T2 were 1.59±0.11, 2.02±0.08 and 1.06±0.17, significantly higher than those in the other groups ( P<0.05). HE staining showed that the implanted hydrogel was tightly bound to the bone tissue. Immunohistochemical staining showed that the numbers of Runx2 and osteocalcin positive cells in group T2 were significantly higher than those in the other groups. Conclusions:With ideal biocompatibility, hydrogel scaffolds with different concentrations of laponite can slowly release the decomposed ions of lithium, magnesium and silicon to promote the osteogenic differentiation of BMSCs and the repair of bone defects in vivo. A 2% concentration of laponite in the hydrogel scaffolds may result in the best results.

Chinese Journal of Endemiology ; (12): 613-618, 2022.
Article in Chinese | WPRIM | ID: wpr-955757


Objective:To investigate the combined effect of fluoride exposure and low nutrition on osteogenesis and osteoclastic differentiation in rats.Methods:SD rats were divided into four groups by the method of random number table, namely normal nutrition group, low nutrition treatment group, fluoride exposure group and co-treatment of fluoride and low nutrition group according to 2 × 2 factorial experimental design, 8 rats in each group, half male and half female. Five months after the experiment, immunohistochemistry was used to test the expression levels of femoral alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteoprotegerin (OPG) and receptor activator of nuclear factor kappa B ligand (RANKL). Analysis of variance of factorial design was used to determine the interaction between fluoride exposure and low nutrition on osteogenesis and osteoclastic differentiation.Results:The immunohistochemical results of bone tissue showed that there were significant differences in the expression levels of osteogenesis differentiation markers ALP and Runx2 between different groups ( F = 25.98, 17.77, P < 0.001). Compared with normal nutrition group (0.005 2 ± 0.002 7, 0.003 1 ± 0.001 4), the expression levels of ALP and Runx2 in fluoride exposure group were higher (0.019 5 ± 0.005 0, 0.014 4 ± 0.004 4, P < 0.05). There was no significant difference between low nutrition treatment group (0.002 6 ± 0.001 8, 0.004 4 ± 0.003 2) and co-treatment of fluoride and low nutrition group (0.003 6 ± 0.000 7, 0.002 9 ± 0.000 8, P > 0.05). The expression levels of ALP and Runx2 in co-treatment of fluoride and low nutrition group were lower than those of fluoride exposure group ( P < 0.05). There were significant differences in the expression level osteoclastic differentiation marker of RANKL and the ratio of RANKL/OPG ( F = 10.50, 31.05, P < 0.001). Among them, the RANKL/OPG ratio (0.115 3 ± 0.039 5) in fluoride exposure group was lower than that in normal nutrition group (1.426 3 ± 0.777 2), and the RANKL expression level and RANKL/OPG ratio (0.019 5 ± 0.007 7, 7.258 7 ± 3.674 3) in co-treatment of fluoride and low nutrition group were higher than those in normal nutrition group (0.004 4 ± 0.002 5, 1.426 3 ± 0.777 2, P < 0.05). However, there was no significant difference in the RANKL expression level and RANKL/OPG ratio (0.004 0 ± 0.001 9, 2.022 3 ± 0.753 7) in low nutrition treatment group ( P > 0.05). The expression level of RANKL and the ratio of RANKL/OPG in the co-treatment of fluoride and low nutrition group were higher than those in low nutrition treatment group and fluoride exposure group ( P < 0.05). The 2 × 2 analysis of variance of factorial design showed that fluoride exposure and low nutrition had interaction on ALP, Runx2, RANKL expression levels and RANKL/OPG ratio ( F = 4.38, 19.39, 22.12, 108.00, P < 0.05), antagonistic effect on ALP and Runx2 expression, synergistic effect on RANKL expression and RANKL/OPG ratio. Conclusions:In rat bone tissue, fluoride exposure promotes osteogenesis differentiation, inhibits osteoclastic differentiation dominated by active osteogenic function. The interaction between fluoride and low nutrition on osteogenesis and osteoclastic differentiation is antagonistic osteogenesis differentiation and synergistic promotion of osteoclastic differentiation. Normal nutrition conditions are material basis of osteogenesis differentiation, and low nutrition is the inducement of enhanced osteoclastic differentiation.

Article in Chinese | WPRIM | ID: wpr-954797


The growth plate (cartilage tissue) is the key to bone development and linear growth.However, as the adolescence proceeds, the proliferation capacity of the growth plate will be continuously consumed, and finally the growth plate will be closed.A variety of regulatory factors control chondrocyte proliferation and differentiation through different mechanisms.Endocrine regulators (including growth hormone, insulin-like growth factor, thyroxine, sex hormone, glucocorticoid, etc.) and transcription factors play an important role in regulating the development of growth plates through systematic modulation.In addition, such local regulatory factors as Indian hedgehog protein, parathyroid hormone-related peptide, bone morphogenetic protein and fibroblast growth factor also regulate the development of the growth plate.In this paper, the regulatory mechanism for chondrocyte proliferation and differentiation was summarized.