Specific RNA m6A modification sites in bone marrow mesenchymal stem cells from the jawbone marrow of type 2 diabetes patients with dental implant failure / 国际口腔科学杂志·英文版

The failure rate of dental implantation in patients with well-controlled type 2 diabetes mellitus (T2DM) is higher than that in non-diabetic patients. This due, in part, to the impaired function of bone marrow mesenchymal stem cells (BMSCs) from the jawbone marrow of T2DM patients (DM-BMSCs), limiting implant osseointegration. RNA N6-methyladenine (m6A) is important for BMSC function and diabetes regulation. However, it remains unclear how to best regulate m6A modifications in DM-BMSCs to enhance function. Based on the "m6A site methylation stoichiometry" of m6A single nucleotide arrays, we identified 834 differential m6A-methylated genes in DM-BMSCs compared with normal-BMSCs (N-BMSCs), including 43 and 790 m6A hypermethylated and hypomethylated genes, respectively, and 1 gene containing hyper- and hypomethylated m6A sites. Differential m6A hypermethylated sites were primarily distributed in the coding sequence, while hypomethylated sites were mainly in the 3'-untranslated region. The largest and smallest proportions of m6A-methylated genes were on chromosome 1 and 21, respectively. MazF-PCR and real-time RT-PCR results for the validation of erythrocyte membrane protein band 4.1 like 3, activity-dependent neuroprotector homeobox (ADNP), growth differentiation factor 11 (GDF11), and regulator of G protein signalling 2 agree with m6A single nucleotide array results; ADNP and GDF11 mRNA expression decreased in DM-BMSCs. Furthermore, gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses suggested that most of these genes were enriched in metabolic processes. This study reveals the differential m6A sites of DM-BMSCs compared with N-BMSCs and identifies candidate target genes to enhance BMSC function and improve implantation success in T2DM patients.

Preliminary study on cerebrospinal fluid proteomics of Erxian Decoction against neurogenesis impairment in late-onset depression / 中国中药杂志

This study aims to elucidate the underlying mechanism of Erxian Decoction(EXD) against neurogenesis impairment in late-onset depression(LOD) rats based on cerebrospinal fluid(CSF) proteomics. A total of 66 20-21-month-old male Wistar rats were randomized into naturally aged(AGED) group, LOD group, and EXD group. All rats received chronic unpredictable mild stress(CUMS) for 6 weeks for LOD modeling except for the AGED group. During the modeling, EXD group was given EXD(ig, twice a day at 4 g·kg~(-1)) and other groups received equivalent amount of normal saline(ig). After modeling, a series of behavioral tests, such as sucrose preference test(SPT), open-field test(OFT), forced swimming test(FST), and Morris water maze test(MWMT) were performed. Immunofluorescence method was used to detect the number of Ki-67/Nesti-positive cells and BrdU/DCX-positive cells in the hippocampal DG area of each group. High-concentration corticosterone(CORT) was combined with D-galactose(D-gal) to simulate the changes of LOD-related stress and aging and the proliferation and differentiation of primary neural stem cells of hippocampus in each group were observed. Data independent acquisition(DIA)-mass spectrometry(MS) was used to analyze the differential proteins in CSF among groups and bioinformatics analysis was performed to explore the biological functions of the proteins. Behavioral tests showed that sucrose consumption in SPT, total traveling distance in OFT, and times of crossing the platform in MWMT were all reduced(P<0.01) and the immobility time in FST was prolonged(P<0.01) in the LOD group compared with those in the AGED group, suggesting that LOD rats had developed depression symptoms such as anhedonia, decreased locomotor activity ability, and cognitive dysfunction. Behavioral abnormalities were alleviated(P<0.01, P<0.05) in the EXD group as compared with those in the LOD group. Immunofluorescence results demonstrated that Ki-67/Nesti-positive cells and BrdU/DCX-positive cells in the hippocampal DG area were fewer(P<0.05) in LOD group than in the AGED group, and the positive cells in the EXD group were more(P<0.05) than those in the LOD group. In vitro experiment showed that the proliferation and differentiation of primary hippocampal neural stem cells under the CORT+D-gal treatment were reduced(P<0.01). The proliferation rate of neural stem cells decreased(P<0.05) in CORT+D-gal+LOD-CSF group but increased(P<0.01) in CORT+D-gal+EXD-CSF group compared with that in the CORT+D-gal group. A total of 2 620 proteins were identified from rat CSF, with 135 differential proteins between the LOD group and AGED group and 176 between EXD group and LOD group. GDF11, NrCAM, NTRK2, and GhR were related to neurogenesis and 39 differential proteins were regulated by both LOD and EXD. EXD demonstrated obvious anti-LOD effect, as it improved the locomotor activity ability and cognitive function of LOD rats and protected the proliferation and differentiation of hippocampal neural stem cells. EXD exerts anti-LOD effect by regulating the proteins related to neurogenesis in CSF, such as GDF11, NrCAM, NTRK2, and GhR and maintaining hippocampal neurogenesis.

GDF11 ameliorated myocardial ischemia reperfusion injury by antioxidant stress and up-regulating autophagy in STZ-induced type 1 diabetic rats

Abstract Purpose: To investigate whether GDF11 ameliorates myocardial ischemia reperfusion (MIR) injury in diabetic rats and explore the underlying mechanisms. Methods: Diabetic and non-diabetic rats subjected to MIR (30 min of coronary artery occlusion followed by 120 min of reperfusion) with/without GDF11 pretreatment. Cardiac function, myocardial infarct size, creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), superoxide dismutase (SOD) 15-F2tisoprostane, autophagosome, LC3II/I ratio and Belcin-1 level were determined to reflect myocardial injury, oxidative stress and autophagy, respectively. In in vitro study, H9c2 cells cultured in high glucose (HG, 30mM) suffered hypoxia reoxygenation (HR) with/without GDF11, hydrogen peroxide (H2O2) and autophagy inhibitor 3-methyladenine (3-MA) treatment, cell injury; oxidative stress and autophagy were assessed. Results: Pretreatment with GDF11 significantly improved cardiac morphology and function in diabetes, concomitant with decreased arrhythmia severity, infarct size, CK-MB, LDH and 15-F2tisoprostane release, increased SOD activity and autophagy level. In addition, GDF11 notably reduced HR injury in H9c2 cells with HG exposure, accompanied by oxidative stress reduction and autophagy up-regulation. However, those effects were completely reversed by H2O2 and 3-MA. Conclusion: GDF11 can provide protection against MIR injury in diabetic rats, and is implicated in antioxidant stress and autophagy up-regulation.

Effects of Low-Dose Decitabine on Soluble CD44, GDF11 Levels and Hematopoietic Function in Elderly Patients with MDS / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of low-dose decitabine on levels of soluble CD44 and GDF11, and hematopoietic function in elderly patients with myelodysplastic syndrome (MDS).@*METHODS@#Ninety-nine patients with senile myelodysplastic syndrome (MDS) admitted to our hospital from October 2015 to October 2017 were divided into group A, B and C according to their treatment, each with 33 cases.The patients in group A were treated with low-dose decitabine, the patients in group B were treated with usual dose of decitabine, and the patients in group C were treated with low-dose decitabine plus G-GSF, cytarabine, and aclarithromycin. The changes of soluble CD44, GDF11 levels and hematopoietic function (sTfR/E) were compared before and after treatment. The clinical remission rate and adverse reaction rate in 3 groups were analyzed.@*RESULTS@#Before treatment, the levels of CD44, GDF11 and sTfR/E were not significantly different between the 3 groups (P＞0.05). After treatment, the levels of CD44 and GDF11 were significantly decreased in these groups, while the serum levels of sTfR/E were significantly increased, and there was no significant difference between the 3 groups (P＞0.05). After treatment, the total effective rates of A, B, and C 3 group were 82.3%, 81.8%, and 78.8%, respectively, without statistically significant difference (P＞0.05). During the treatment, the incidence of non-hemotoxic adverse reactions in group A was 8.8%, significantly lower than that in group B and C (30.3%, 27.3%) (P＜0.05, P＜0.05), the incidence of hemotoxic adverse reactions in group A was 39.4%, significantly lower than that 63.6% and 66.7% in group B and C (P＜0.05, P＜0.05).@*CONCLUSION@#Low-dose decitabine alone is effective in treating elderly patients with MDS as compared with conventional dose and combination therapy, moreover can significantly reduce the levels of CD44 and GDF11, improve hematopoietic function and low the adverse reactions. Thereby the low dose of decitabine may be a new choice for clinical treatment of MDS.

Substitutos ósseos sintéticos na Implantodontia / Synthetic bone substitutes for Implant Dentistry ­ a literature review

Objetivo: analisar sistematicamente, na literatura científica, o uso de substitutos ósseos sintéticos na regeneração óssea para Implantodontia. Material e métodos: uma busca foi realizadas nas bases de dados PubMed, Cochrane, LILACS e SciELO, nos últimos quinze anos, combinando as palavras-chave "regeneração óssea", "materiais biocompatíveis", "implantes dentários", e "materiais aloplásticos". Resultados: Dos 199 artigos inicialmente recuperados, apenas 27 foram selecionados, incluindo revisões sistemáticas/ metanálises (n=2), revisões da literatura (n=1), estudos clínicos (n=9) e pré-clínicos (n=12), relato de caso (n=1) e tese (n=1). Nos modelos animais, o vidro bioativo é capaz de provocar formação óssea à distância, inibir a migração apical do epitélio juncional, e gerar maior deposição de cemento na superfície radicular. Partículas esféricas geram melhor dissolução e integração com o novo osso circundante, e bons resultados são vistos nas técnicas de ROG e RTG. Em um estudo clínico randomizado, o vidro bioativo misturado ao osso autógeno para regeneração de defeitos intraósseos reduziu significativamente a profundidade de sondagem, com ganho de nível clínico de inserção, e resolução dos defeitos já aos seis meses. Nos modelos animais, a tríade hidroxiapatita (HA), beta-fosfato tricálcio (ß-TCP), e fosfato de cálcio bifásico (HA+ ß-TCP) tem se mostrado biocompatível, biorreabsorvível e osteocondutora. Um estudo clínico controlado com HA+ ß-TCP/ membrana revelou melhor preservação óssea vertical e horizontal comparado ao coágulo/membrana, nas TCFCs de seis meses. Regenerações ósseas verticais significativas com estes materiais sintéticos são potencializadas pelo uso dos fatores de crescimento. Conclusão: substitutos ósseos sintéticos demonstram uso promissor para regeneração. Entretanto, a evidência clínica deve ser substancialmente aumentada.

Objective: to systematically analyze in the scientifi c literature the use of synthetic bone substitutes for bone regeneration in implant dentistry. Material and methods: a search was conducted at the PubMed, Cochrane, LILACS and SciELO databases considering the last fi fteen years, and combining the keywords "bone regeneration", "biocompatible materials", "dental implants", and "alloplastic materials". Results: of the 199 articles initially retrieved, only 27 were selected, including systematic reviews/meta-analysis (n=2), literature reviews (n=1), clinical (n=9) and pre-clinical (n=12) studies, case report (n=1) and thesis (n=1). In animal models, the bioactive glass can cause bone formation at distance, inhibit apical migration of the junctional epithelium, and generate greater deposition of cementum over the root surface. Spherical particles generate better dissolution and integration with the new surrounding bone, and good results are seen in the ROG and RTG techniques. In a randomized study, the bioactive glass mixed with autogenous bone to regenerate intra-osseous defects signifi cantly reduced probing depths, with clinical attachment level gains, and defect resolution as early as 6 months. In animal models, the triad hydroxyapatite (HA), beta-tricalcium phosphate (ß-TCP), and biphasic calcium phosphate (HA + ß-TCP) has been shown to be biocompatible, bioresorbable, and osteoconductive. A controlled clinical study with (HA + ß-TCP/membrane) showed better vertical and horizontal bone preservation compared to clot / membrane at the 6 month CBCT images. Also, signifi cant vertical bone regeneration with these synthetic materials is enhanced by the use of growth factors. Conclusion: the synthetic bone substitutes are good candidates for regeneration. However, the level of clinical evidence must be substantially increased.

Construction of a recombinant adenovirus co-expressing bone morphogenic proteins 9 and 6 and its effect on osteogenesis in C3H10 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus co-expressing bone morphogenic protein (BMP) 9 and BMP6 and observe its effect on the osteogenesis in C3H10 cells.</p><p><b>METHOD</b>The full-length sequences of BMP9 and BMP6 were amplified from AdEasy vector by PCR and cloned into the shuttle plasmid pASG2 vector to construct the co-expression shuttle plasmid pASG2-BMP9, 6 followed by homologous recombination with plasmid pAdeasy-1 in BJ5183. After confirmation by restriction endonuclease digestion, the recombinant vector was transfected into HEK293 cells, and high-titer recombinant adenovirus (Ad-BMP9, 6) was collected after amplification. Ad-BMP9, 6 was then transduced into C3H10 cells in vitro, and the mRNA expression of BMP9 and BMP6 was detected by RT-PCR. The osteogenic capability of the transfected cells was observed by alkaline phosphatase staining and calcium-alizarin red staining.</p><p><b>RESULTS</b>AdBMP9,6 was constructed successfully and effectively infected in C3H10 cells, in which high expressions of BMP6 and BMP9 were detected. C3H10 cells infected with Ad-BMP9,6 showed stronger alkaline phosphatase and calcium-alizarin red staining than the cells transfected by either BMP9 or BMP6 alone.</p><p><b>CONCLUSION</b>The recombinant adenovirus co-expressing BMP9 and BMP6 we constructed shows a more potent effect than the adenoviruses expressing either BMP9 or BMP6 alone in inducing the osteogenic differentiation of C3H10 cells into osteoblasts.</p>

Orientações para uso clínico da BMP / Orientations for clinical use of BMP

Qual é a origem das BMPs e qual a vasta sinonímia empregada para identificá-las?As proteínas morfogenéticas ósseas são as responsáveis pela sinalização para a indução da formação óssea. As BMPs (bone morphogenetic proteins) representam uma família com mais de vinte proteínas relatadas, as quais fazem parte da família dos fatores ß de crescimento e transformação (TGF-ß), ativando e inibindo os fatores de diferenciação e crescimento (GDFs).

What is the origin of BMPs and how extensive is the synonymy employed to identify them?The morphogenetic proteins of the bone are responsible for signaling to induce bone formation. BMPs (bone morphogenetic proteins) represent a family with more than 20 reported proteins, which are part of the transforming growth ß factor family (TGF-ß), activating and inhibiting the differentiation and growth factors (GDF).

Effect of human bone morphogenetic protein 2, 3, 6, and 12 on osteosarcoma cell line UMR106 / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of human bone morphogenetic protein (hBMPs) 2/3/6 and 12 on osteosarcoma cell UMR106.@*METHODS@#Adenovirus-BMP2/3/6 and 12 (AdBMP2/3/6 and12) were used to treat the cell line. Their proliferation, apoptosis, and transmigration were detected by Trypan blue exclusion test, TdT-mediated biotinylated-dUTP nick end labeling (TUNEL), acridine orange-ethidium bromide (AO/EB) double fluorescent dye staining, and transwell-room test, respectively. The alkaline phosphatase (ALP) activity was detected to reflect the differentiation of tumors.@*RESULTS@#Compared with the control groups, the cell survival rate of the experimental groups treated with AdBMP2/3/6 and 12 showed a significant time-dependent decrease (P<0.01). The apoptosis indexes were increased significantly (P<0.01) and the results from TUNEL and AO/EB method were consistent. The cell numbers of transmembrane significantly decreased at 24,48, and 72 h (P<0.01). AdBMP2/3/6 and 12 treatment enhanced the activity of ALP activity from day 3 and this effect might still be observed up to day 9 of the treatment (P<0.01).@*CONCLUSION@#hBMPs2/3/6 and 12 can inhibit the proliferation and transmigration, and induce their apoptosis and differentiation in osteosarcoma cell line UMR106.