Effect of walking with blood flow restriction in elderly women with osteoporosis/osteopenia / Efeito da caminhada com restrição do fluxo sanguíneo em idosas com osteoporose/osteopenia

Abstract Introduction The preservation of bone mass in elderly women is associated with better levels of practice of systematic physical exercises. Aerobic training combined with blood flow restriction seems to be a new alternative that determines this process, but knowledge gaps are still observed when referring to exercise associated with blood flow restriction (BFR) and adaptations on bone variables. Objective To analyze the chronic effects of aerobic training with and without BFR on bone mineral density and bone biomarker osteocalcin concentrations in older women. Methods Thirty women were randomized into the following groups: walking on a treadmill at low intensity with BFR; moderate treadmill walking with no BFR; only BFR (no exercise) for 20 minutes, twice a week, for 24 weeks. Bone mineral density was measured before and 24 weeks after intervention. Blood serum osteocalcin concentrations were measured before, 12 and 24 weeks after intervention. Results There were no differences between groups in bone mineral density (femoral neck, p = 0.31; total femur, p = 0.17; lumbar spin, p = 0.06) and osteocalcine (W(2) = 0.27; p = 0.87) ouctomes after 24 weeks of intervention. Conclusion There was no difference between walking training, blood flow restriction only, or walking+blood flow restriction on bone mineral density and osteocalcin concentrations after 24-weeks of intervention in older women with osteopenia/osteoporosis.

Resumo Introdução A preservação da massa óssea em mulheres idosas está associada a melhores níveis de prática de exercícios físicos sistemáticos. O treinamento aeróbico combinado com restrição de fluxo sanguíneo (RFS) parece ser uma nova alternativa que determina esse processo, mas ainda são observadas lacunas de conhecimento quando se refere ao exercício associado à RFS e adaptações nas variáveis ósseas. Objetivo Analisar os efeitos crônicos do treinamento aeróbico com e sem RFS na densidade mineral óssea e nas concentrações do biomarcador ósseo osteocalcina em mulheres idosas. Métodos Trinta mulheres foram randomizadas nos seguintes grupos: caminhada em esteira de baixa intensidade com RFS; caminhada moderada em esteira sem RFS; apenas RFS (sem exercícios) por 20 minutos, duas vezes por semana, durante 24 semanas. A densidade mineral óssea foi medida antes e 24 semanas após a intervenção. As concentrações séricas de osteocalcina no sangue foram medidas antes, 12 e 24 semanas após a intervenção. Resultados Não houve diferenças entre os grupos na densidade mineral óssea (colo do fêmur, p = 0,31; fêmur total, p = 0,17; giro lombar, p = 0,06) e osteocalcina (W(2) = 0,27; p = 0,87) após 24 semanas de intervenção. Conclusão Não houve diferença entre treinamento de caminhada, apenas restrição de fluxo sanguíneo ou caminhada + restrição de fluxo sanguíneo na densidade mineral óssea e nas concentrações de osteocalcina após 24 semanas de intervenção em mulheres idosas com osteopenia/osteoporose.

Regulation of osteogenic differentiation under hypoxia by EphrinB2/EphB4 in MC3T3-E1 cells / 口腔疾病防治

Objective @# To investigate the effect of erythropoietin producing hepatocyte kinase receptor ligand B2-erythropoietin producing hepatocyte kinase receptor B4 (EphrinB2/EphB4) on the osteogenic differentiation of MC3T3-E1 cells in a hypoxic environment to provide experimental evidence for hypoxia regulation of osteoblast differentiation.@*Methods @# Control groups and cobalt chloride (CoCl2)-induced hypoxia groups were set up first. qRT-PCR was used to detect the mRNA expression of the osteogenic markers alkaline phosphatase (ALP), collogen1 (COL I), runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN). ALP staining was used to detect the activity of cell alkaline phosphatase after osteogenic induction. The mRNA and protein expression levels of hypoxia inducible factor-1α (HIF-1α), EphrinB2 and EphB4 in the two groups were detected via qRT-PCR and Western blot. Then, the CoCl2 + inhibitor group was established. NVP-BHG712, an EphB4 phosphorylation inhibitor, was added to this group to prevent EphrinB2 from binding to EphB4 and producing signals. qRT-PCR and Western blot were used to detect the mRNA and protein expression of osteogenic markers, including ALP, RUNX2, COL I, and OCN. ALP staining and Alizarin red S staining were used to measure osteoblast differentiation and mineralization. @*Results @# Compared with the control group, the mRNA expression of the osteogenic differentiation markers ALP, RUNX2, COL-1, and OCN in MC3T3-E1 cells increased, and ALP activity and mineralization were enhanced under CoCl2-induced hypoxia in vitro (P<0.05). Additionally, the expression of HIF-1α, EphrinB2 and EphB4 was upregulated at the mRNA and protein levels under hypoxia (P<0.05). When NVP-BHG712 was used to block the connection between EphrinB2 and EphB4, the expression of osteogenic markers and ALP activity and mineralization were decreased (P<0.05).@*Conclusion@#EphrinB2/EphB4 can promote osteogenic differentiation of MC3T3-E1 cells and increase the expression of osteogenic markers and tissue mineralization in a hypoxic environment.

Expressions of serum N-terminal osteocalcin and cytokeratin 5/6 in primary lung cancer patients with bone metastasis and their clinical significances / 肿瘤研究与临床

Objective:To explore the expressions of serum N-terminal osteocalcin (N-MID) and cytokeratin (CK) 5/6 in primary lung cancer patients with bone metastasis and their clinical significances.Methods:The clinical data of 96 patients with primary lung cancer admitted to Chengdu Second People's Hospital between February 2019 to February 2022 were retrospectively analyzed. All patients were divided into the bone metastasis group (38 cases) and the non-bone metastasis group (58 cases) according to whether bone metastasis occurred, and 45 healthy people who underwent physical examination during the same period were treated as the healthy control group. The expression levels of serum N-MID and CK5/6 in the bone metastasis group, the non-bone metastasis group and the healthy control group were compared. Logistic regression was used to analyze the factors affecting bone metastasis in patients with primary lung cancer; receiver operating characteristic (ROC) curve analysis was used to analyze the value of the expression levels of serum N-MID and CK5/6 in predicting bone metastasis in patients with primary lung cancer.Results:The composition ratio of patients with pathological stage Ⅲ-Ⅳ, serum bone-derived alkaline phosphatase and N-MID expression levels in the bone metastasis group were higher than those in the non-bone metastasis group (all P < 0.05). The expression level of serum N-MID in the bone metastasis group and non-bone metastasis group was higher than that in the healthy control group [(26.0±5.3) ng/ml, (15.3±3.1) ng/ml vs. (9.9±1.7) ng/ml, F = 224.27, P < 0.001], and there were statistically significant differences in the serum N-MID expression level of the pairwise comparison among the three groups (all P < 0.05). The expression level of serum CK5/6 in the bone metastasis group and the non-bone metastasis group was lower than that in the healthy control group [(3.6±0.7) ng/ml, (7.3±1.4) ng/ml vs. (10.6±2.4) ng/ml, F = 178.11, P < 0.001], and there were statistically significant differences in the serum CK5/6 expression level of the pairwise comparison among the three groups (all P < 0.05). Multivariate analysis showed that CK5/6, N-MID and bone-derived alkaline phosphatase were independent affecting factors for bone metastasis in patients with primary lung cancer ( OR = 4.088, 3.615, 2.892, all P < 0.05). ROC curve analysis showed that the optimal cut-off values of serum N-MID and CK5/6 expression levels for predicting bone metastasis in patients with primary lung cancer were 18.59 ng/ml and 4.71 ng/ml; the corresponding the area under the curve (AUC) was 0.881 and 0.862, respectively; and the specificity and AUC of the combination of serum N-MID and CK5/6 in predicting the bone metastasis in patients with primary lung cancer was 98.28% and 0.937 (95% CI 0.869-0.977), respectively; the AUC predicted by the combination of both was higher than that by serum N-MID or CK5/6 single (all P < 0.001). Conclusions:The expression levels of serum N-MID and CK5/6 in primary lung cancer patients with bone metastasis are abnormally changed. Clinical detection of serum N-MID and CK5/6 expression levels may be used as sensitive indicators for predicting the bone metastasis in patients with primary lung cancer.

Clinical effects of Jiakang Pingxiao prescription combined with methiimidazole on hyperthyroidism / 中国基层医药

Objective:To investigate the clinical effects of Jiakang Pingxiao prescription combined with methiimidazole on hyperthyroidism. Methods:A total of 100 patients with hyperthyroidism admitted to Shanxian Central Hospital from February 2018 to January 2021 were included in this study. They were randomly divided into a study group and a control group, with 50 patients in each group. The control group was treated with methiimidazole, and the study group was treated with Jiakang Pingxiao prescription combined with methiimidazole. Thyroid function, serum levels of osteocalcin (OCN), β-CTx, hypersensitive C-reactive protein, and interleukin-6 (IL-6) were compared between the two groups. Results:After treatment, serum levels of thyroid stimulating hormone (TSH), free triiodothyronine (FT3), free thyroxine (FT4) in the study group were (3.10 ± 1.36) mU/L, (5.76 ± 1.25) pmol/L, (15.22 ± 1.95) pmol/L, respectively, which were significantly lower than (4.88 ± 1.47) mU/L, (7.13 ± 1.32) pmol/L, (19.07 ± 2.02) pmol/L in the control group ( t = 5.27, 4.71, 6.29, all P < 0.05). Serum OCN, β-CTx, hS-CRP, and IL-6 in the study group were (17.36 ± 2.62) μg/L, (0.32 ± 0.04) μg/L, (4.07 ± 0.86) mg/L, and (1.38 ± 0.21) pg/L, respectively, which were significantly lower than (26.05 ± 2.88) μg/L, (0.51 ± 0.09) μg/L, (6.23 ± 0.91) mg/L, (1.89 ± 0.28) pg/L in the control group ( t = 12.37, 10.40, 7.39, 8.57, all P < 0.05). The incidence of adverse reactions in the study group was significantly lower than that in the control group [6.00% (3/50) vs. 12.00% (3/50), χ2 = 14.78, P < 0.05). Conclusion:Jikang Pingxiao prescription combined with methiimidazole can effectively reduce the inflammatory responses in patients with hyperthyroidism, inhibit the expression of OCN and β-CTX in the serum, and improve thyroid function. The combined method is scientific and reasonable, and is suitable for clinical application. It has good therapeutic effects on hyperthyroidism and is worthy of clinical promotion.

Effects of teriparatide on bone formation in rats with experimentally induced premaxillary expansion

ABSTRACT Objective: This study aimed to evaluate the effects of systemic teriparatide on sutural bone formation after premaxillary suture expansion in rats. Material and Methods: Twenty Wistar male rats (8-10 weeks old) were randomly divided into two groups, namely, control (C, n=10) and teriparatide (T, n=10). An expansion force was applied to the maxillary incisors using helical spring for a seven-day expansion period, for both groups. On the eighth day, the rats were kept for a seven-day consolidation period, and then 60 µg/kg teriparatide (once a day) was administered to group T subcutaneously for seven days. Then, all the rats were sacrificed, and histological sections were stained with hemotoxylin-eosin for examination. Anti-osteonectin, anti-osteocalcin, anti-Vascular endothelial growth factor (VEGF) and anti-transforming growth factor beta (TGF-β) were evaluated by immunohistochemical analysis in the midpalatal suture area. Results: Histologically, the newly formed bone tissue was observed to be larger in group T than in group C. The number of immunoreactive osteoblasts for osteonectin, osteocalcin and VEGF antibodies was significantly higher in group T than in group C (p = 0.0001). The TGF-β antibody showed a mild reaction in group T, but did not reach significance in comparison with group C (p ˃ 0.05). Conclusion: Systemic teriparatide application following the premaxillary expansion of the suture area may stimulate bone formation and add to the consolidation of the expansion in rats by regulating osteonectin, osteocalcin and VEGF.

RESUMO Objetivo: O presente estudo teve como objetivo avaliar os efeitos do uso sistêmico da teriparatida na formação óssea sutural após a expansão da pré-maxila em ratos. Material e Métodos: Vinte ratos machos da raça Wistar (com oito a dez semanas de vida) foram divididos aleatoriamente em dois grupos: controle (C, n=10) e teriparatida (T, n=10). Uma força de expansão foi aplicada aos incisivos superiores, usando uma mola helicoidal, por um período de expansão de sete dias em ambos os grupos. No oitavo dia, os ratos iniciaram um período de sete dias de consolidação, nos quais 60 µg/kg de teriparatida foram administrados (uma vez ao dia), por via subcutânea, para o grupo T. Posteriormente, todos os ratos foram sacrificados e cortes histológicos corados com hemotolixina-eosina foram examinados. Por meio de análise imuno-histoquímica da região da sutura palatina mediana, avaliou-se a presença de anti-ostenectina, anti-osteocalcina, anti-fator de crescimento endotelial vascular (VEGF) e anti- fator transformador de crescimento (TGF-β). Resultados: Histologicamente, observou-se que o tecido ósseo recém-formado foi maior no grupo T do que no grupo C. O número de osteoblastos imunorreativos para anticorpos de osteonectina, osteocalcina e VEGF foi significativamente maior no grupo T do que no grupo C (p = 0,0001). O anticorpo TGF-β mostrou uma pequena reação no grupo T; porém, sem diferença significativa para o grupo C (p ˃ 0,05). Conclusão: O uso sistêmico de teriparatida após a expansão da sutura na região da pré-maxila pode estimular a formação óssea e melhorar a consolidação da expansão em ratos, por meio da regulação de osteonectina, osteocalcina e VEGF.

CCDC134 regulates the osteogenic differentiation of human dental pulp stem cells / 口腔疾病防治

Objective @#To study the regulatory effect of coiled-coil domain containing 134 (CCDC134) on the osteogenic differentiation of human dental pulp stem cells (hDPSCs).@*Methods @# HDPSCs were isolated and cultured from dental pulp tissue and transfected with NC-CCDC134, shCCDC134 and CCDC134 lentiviruses. They were divided into the control group, negative control group, CCDC134 downregulation (shCCDC134) group and CCDC134 overexpression (CCDC134) group. Surface markers of hDPSCs (Stro-1, CD105, CD34, CD45) were detected by flow cytometry; colony formation was analyzed by toluidine blue staining; ALP expression was estimated by ALP staining; mineralized nodule formation was evaluated by alizarin red staining; lipid droplet formation was examined by oil red staining; and gene and protein expression of CCDC134, Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), bone morphogenetic protein-2 (BMP-2), and mothers against decapentaplegic homolog 1 (SMAD1) was detected by qPCR and western blot, respectively. Further, a BMP-2 activator (BMP-2) and inhibitor (Dorsomorphin) were used to down-regulate and up-regulate CCDC134, respectively (shCCDC134, shCCDC134+BMP-2, CCDC134, CCDC134+Dorsomorphin), in hDPSCs. The hDPSC aggregates were subcutaneously transplanted into nude mice for 2 months, and new bone formation was detected by H&E staining. The BMP-2/SMAD1 signaling in each group was detected by qPCR.@*Results@#hDPSCs showed high expression of mesenchymal markers and low expression of hematopoietic markers. Compared with the control group, the expression of CCDC134 was increased in the osteogenic-induced hDPSCs (P < 0.05). Compared with the negative control group, the expression of CCDC134 was decreased in the shCCDC134 group, whereas it was increased in the CCDC134 group (P < 0.05). The mineralized nodules, osteogenic genes and proteins in the shCCDC134 group were decreased (P < 0.05), while they were increased in the CCDC134 group (P < 0.05). The expression of BMP-2/SMAD1 signaling decreased in the shCCDC134 group, while it increased in the CCDC134 group (P < 0.05). Compared to the shCCDC134 group, osteogenic genes and proteins increased in the shCCDC134+BMP-2 group, and subcutaneous new bone formation increased in nude mice (P < 0.05). The indexes of the CCDC134+Dorsomorphin group decreased compared with the CCDC134 group (P < 0.05).@*Conclusion@#CCDC134 promotes the osteogenic differentiation of hDPSCs by regulating the BMP-2/SMAD1 signaling pathway.

Association between dental fluorosis and serum biomarkers of bone metabolism in children / 中华地方病学杂志

Objective:This study was aimed to explore the associations between the risk of dental fluorosis and the serum biomarkers of bone metabolism in children.Methods:A total of 502 children aged 7 - 12 years were selected by cluster sampling from 4 primary schools in Tongxu County, Kaifeng City, Henan Province from April to May 2017. Morning urine and fasting peripheral blood samples were collected from each participant. Urinary fluoride concentration was determined by fluoride ion selective electrode method. Serum calcium, phosphorus, alkaline phosphatase (ALP), bone-specific alkaline phosphatase (BALP), osteocalcin (OC), calcitonin (CT) and parathyroid hormone (PTH) levels were measured using an automatic biochemical analyzer. Dean method was used to evaluate the prevalence of dental fluorosis in children, and the participants were divided into dental fluorosis group ( n = 173) and control group ( n = 329) after being diagnosed by trained physicians for their dental fluorosis. The associations between the risk of dental fluorosis and the serum biomarkers of bone metabolism in children were analyzed by logistic regression. Results:The levels of serum phosphorus (mmol/L: 1.54 ± 0.19 vs 1.58 ± 0.21) and OC (ng/ml: 11.59 ± 5.22 vs 12.78 ± 5.88) in children in dental fluorosis group were significantly lower than those in children in control group ( P < 0.05). Logistic regression analysis showed that serum OC level affected the risk of dental fluorosis [odds ratio ( OR) = 0.96, 95% confidence interval ( CI): 0.92 - 0.99, P < 0.05]. The relative contribution of the biomarkers of bone metabolism to the risk of dental fluorosis in descending order were serum OC (36.34%), phosphorus (25.89%), BALP (13.16%), PTH (9.73%), calcium (9.44%), CT (3.72%) and ALP (1.72%). Conclusions:The prevalence of dental fluorosis in children is related to the changes of serum biomarkers of bone metabolism. Serum OC plays an important role in the occurrence of dental fluorosis.

Serum osteocalcin and diabetic peripheral neuropathy in male patients with diabetes / 中华内分泌代谢杂志

Objective:To investigate the association between serum bone turnover marker osteocalcin and the distal symmetric poly neuropathy(DSPN) in male diabetic patients.Methods:Clinical data from 370 male diabetic patients who admitted to Zhongshan Hospital, Fudan University from January 2020 to November 2021 were collected. These patients were grouped into tertiles by serum osteocalcin level: T1 group(osteocalcin<9.2 ng/mL, n=123), T2 group(osteocalcin 9.2-13 ng/mL, n=122), and T3 group(osteocalcin≥13 ng/mL, n=125). The percentage ratios of DSPN were compared among these groups. Using logistic regression model, the adjusted odds ratio ( OR) for DSPN was calculated. Results:There were 50(40.7%), 29(23.8%), 49(39.2%) patients with DSPN in T1, T2, and T3 group respectively. The ratio of patients with DSPN in osteocalcin T2 group were lower than that in the T1 and T3 groups. Further logistic regression showed a 133.9%( OR=2.339, 95% CI 1.097-4.988, P=0.028) and a 134.2%( OR= 2.342, 95% CI 1.040-5.275, P=0.039) increased risk for DSPN in the T1 and T3 group respectively compared with the T2 group, even after adjusted for age, diabetic duration, HbA 1C, diabetic complications, β cross-linked C-telopeptide of type Ⅰ collagen(CTXβ), 25-hydoxy vitamin D(25-OHD), bone mineral density, and treatment. Conclusions:The serum levels of bone turnover marker osteocalcin were associated with the occurrence of DSPN in male diabetic patients, a moderate level of bone turnover(the serum osteocalcin level of between 9.2 and 13 ng/mL for instance) might be protective for male diabetic patients from DSPN.

Platelet-concentrated and platelet poor-plasma promote different pattern on immunohistochemical expression of TGF-β1, however they impairs the osteoneogensis in calvarial defects treated with autograft due suppression of osteocalcin

ABSTRACT Purpose Herein we evaluated the effects of platelet concentrate (PC) and platelet-poor plasma (PPP) on bone repair using noncritical defects in the calvaria of rabbits and compared them to the presence of TGF-β1 and osteocalcin on reparative sites. Methods Five noncritical defects of 8.7 mm in diameter were created on the calvaria of 15 animals. Each defect was treated differently, using autograft (ABG), ABG associated with PC (ABG + PC), ABG with PPP (ABG + PPP), isolated PPP, and blood clot (control). The animals were submitted to euthanasia on the second, fourth and sixth week post-surgery. Results The defects that received ABG+PC or PPP demonstrated lower bone formation when compared to specimens that received ABG in the same period. These results coincided to significant higher immunopositivity for TGF-β1 for specimens that received PC, and lower presence of cytokine in the group PPP. However, either higher or lower presence of TGF-β1 were also correlated to lower presence of osteocalcin. Likewise, these results were similar to findings in specimens treated only with PPP when compared to control. Conclusions PC and PPP were not effective when applied in association with ABG. Similarly, isolated use of PPP was not beneficial in optimizing the bone repair.

Influência do LED Violeta associado ou não ao gel clareador na inflamação, mineralização e maturação colágena do tecido dentinário e pulpar / Influence of violet LED associated or not with peroxide gel on inflammation, mineralization, and collagen fiber maturation in dentin and pulp tissue

Objetivo: Avaliar a influência do LED Violeta, associado ou não ao gel clareador a base de peróxido de hidrogênio (PH) a 17,5% no complexo dentino-pulpar de ratos. Materiais e métodos: Molares superiores de 80 ratos foram distribuídos nos grupos (n = 10): CONT ­ sem tratamento, PH ­ 1 aplicação de 30 minutos de PH a 17,5%, LED ­ 1 aplicação de 20 minutos do LED Violeta, e PH+LED - aplicação do PH e LED Violeta. Imediatamente (T0), e aos 7 (T1), 15 (T2) e 30 dias (T3) após o tratamento, os ratos foram eutanasiados e as maxilas processadas para avaliação histológica, imunoistoquímica (IL-17, IL-23 e osteocalcina), e de picrosírius red, sendo realizados os testes de Wilcoxon e Mann-Whitney e Teste-T pareado e Teste-T, respectivamente. (α = 0,05). Resultados: Necrose e infiltrado inflamatório severo foram observados nos grupos PH e PH+LED. Apenas o grupo PH+LED manteve a imunomarcação severa para IL-17 e IL-23, diferindo do grupo LED e PH que apresentaram moderada imunomarcação em T0. Os grupos PH e PH+LED apresentaram severa imunomarcação de OCN em T2 e moderada imunomarcação em T3. O grupo LED apresentou menor quantidade de fibras imaturas em T2 e T3 que o grupo CONT. Conclusão: A terapia com LED violeta não induziu inflamação e fibrose no tecido pulpar, apesar de acelerar a maturação das fibras de colágeno da dentina e, quando associada ao peróxido de hidrogênio, pode tornar os dentes mais sensíveis(AU)

Objective: Was to evaluated the influence of the Violet LED, associated or not with a 17.5% hydrogen peroxide (HP) bleaching gel in the dentin-pulp complex of rats. Materials and methods: Upper molars of eighty Wistar rats were distributed in the groups (n = 10): CONT - without treatment, HP - 1 application of 30 minutes of 17.5% HP, LED - 1 application of 20 minutes of the Violet LED, and HP+LED - application of HP and LED Violet. Immediately (T0), and at 7 (T1), 15 (T2) and 30 days (T3) after treatment, the rats were euthanized and the jaws were processed for histological, immunohistochemical evaluation (IL-17, IL-23 and osteocalcin), and picrosirius red, with Wilcoxon and Mann-Whitney tests and paired T-test and T-test, respectively (α = 0.05). Results: Necrosis and severe inflammatory infiltrate were observed in the PH and PH+LED groups. Only the PH+LED group maintained severe immunostaining for IL-17 and IL-23, differing from the LED and PH group which presented moderate T0 immunostaining. The PH and PH+LED groups presented severe immunostaining of OCN in T2 and moderate immunostaining in T3. The LED group had a lower amount of immature fibers in T2 and T3 than the CONT group. Conclusion: Violet LED therapy induced no inflammation and fibrosis in the pulp tissue, however accelerating the maturation of dentin collagen fibers and, when associated with hydrogen peroxide, can make the teeth more sensitive(AU)

Clinical efficacy of methimazole in the treatment of hyperthyroidism / 中国基层医药

Objective:To investigate the clinical efficacy of methimazole in the treatment of hyperthyroidism.Methods:A total of 110 patients with hyperthyroidism who received treatment in Zhuji Central Hospital between January 2016 and June 2019 were included in this study. They were randomly assigned to receive treatment either with propylthiouracil (control group, n = 55) or methimazole (observation group, n = 55) for 6 successive months. Thyroid function indicators, bone metabolism indicators, clinical efficacy, and adverse events were compared between the control and observation groups. Results:After treatment, free triiodothyronine (FT 3), free thyroxine (FT 4) and thyroid stimulating hormone (TSH) levels in the observation group were (4.46 ± 1.02) pmol/L, (14.45 ± 2.16) pmol/L and (1.89 ± 0.64) mU/L respectively, which were significantly lower than those in the control group [(6.37 ± 1.38) pmol/L, (18.54 ± 4.46) pmol/L and (3.47 ± 0.99) mU/L, t = 8.254, 6.121, 9.940, all P < 0.05). Calcitonin level in the observation group was significantly higher than that in the control group [(68.62 ± 6.75) ng/L vs. (61.45 ± 6.47) ng/L, t = 5.687, P < 0.05]. Bone Gla-protein level in the observation group was significantly lower than that in the control group [(6.38 ± 1.64) ng/L vs. (8.21 ± 2.19) ng/L, t = 4.960, P < 0.05]. Total effective rate in the observation group was significantly higher than that in the control group [92.73% (51/55) vs. 78.18% (43/55), χ2= 4.681, P < 0.05]. Adverse reaction rate in the observation group was significantly lower than that in the control group [9.09% (5/55) vs. 25.45% (14/55), χ2= 5.153, P < 0.05]. Conclusion:Methimazole is safe and effective in the treatment of hyperthyroidism, which can effectively improve thyroid function and bone metabolism. This study is of certain clinical significance and innovation.

Kaempferol promotes osteogenic differentiation of mouse bone marrow mesenchymal cells under tension stress via the mTORC1 signaling pathway / 口腔疾病防治

Objective @#To investigate the activation of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway molecules during the process by which kaempferol (Kae) promotes osteogenic differentiation of mouse bone marrow mesenchymal cells (BMMCs) under cyclic and uniaxial tension.@*Methods @#BMMCs isolated and cultured in vitro were subjected to uniaxial dynamic tension with a 10% shape variable. The appropriate concentration of Kae was selected by cytotoxicity testing. The endogenous mTOR signal was inhibited by pp242. Four hours after traction, alkaline phosphatase (ALP) and osteocalcin (OCN) were detected by chemical colorimetry and ELISA, and the relative concentration of intracellular calcium was detected by flow cytometry. Phosphorylation of mTOR, 4E/BP1, and ribosomal protein S6 kinases (S6K), which are the main molecules of the endogenous mTORC1 signaling pathway, and expression of osteogenic transcription factors (Runx2 and Osterix) were detected by western blotting (WB), and mRNA expression levels of the above factors were detected by qRT-PCR.@*Results @# The cytotoxicity test showed that 10 μmol/L Kae had little inhibitory effect on cell proliferation but had the strongest osteogenic ability. Four hours after stretching, Kae effectively promoted the osteogenic differentiation of BMMCs. The expression of ALP was （153.04 ± 18.72） U/mg, the expression of OCN was （1.64 ± 0.25） U. The mRNA and protein levels of Runx2 and Osterix were upregulated, and the intracellular calcium content was decreased. The mRNA and protein phosphorylation of mTOR and S6K was upregulated, and the opposite effect was observed with 4E/BP1. After pp242 was added to inhibit mTOR signaling, mTOR and S6K mRNA and protein phosphorylation were downregulated, but 4E/BP1 mRNA and protein phosphorylation was upregulated. The osteogenic differentiation of BMMCs was also significantly inhibited, mRNA and protein expression of Runx2 and Osterix were significantly downregulated, ALP and OCN expression were downregulated, and intracellular calcium content was increased. @* Conclusion@#Kae promotes osteogenic differentiation of mouse BMMCs under uniaxial dynamic tension through the mTORC1 signaling pathway.

Locking plate external fixation combined with membrane induction technology for the treatment of open and comminuted tibial fractures with bone defects / 中国骨伤

OBJECTIVE@#To explore clinical effect of locking plate external fixation combined with membrane induction technology in treating open and comminuted tibial fractures with bone defects.@*METHODS@#Totally 92 patients of open and comminuted tibial fractures with bone defects were chosen form January 2018 to July 2019, and randomly divided into external fixation group and internal fixation group, 46 patients in each group. In external fixation group, there were 29 males and 17 females, aged from 25 to 62 years old, with an average of (37.45±10.92) years old;according to AO classification, 15 patients were type A, 22 patients were type B and 9 patients were type C;according to Gustilo classification, 21 patients were typeⅡ, 10 patients were type ⅢA, 10 patients were type ⅢB, 5 patients were type Ⅲ C;treated by fracture reduction with locking plate external fixation. In internal fixation group, there were 31 males and 15 females, aged from 23 to 60 years old, with an average of(36.88±10.64) years old;according to AO classification, 18 patients were type A, 20 patients were type B and 8 patients were type C; according to Gustilo classification, 22 patients were typeⅡ, 11 patients were type ⅢA, 7 patients were type ⅢB, 6 patients were type Ⅲ C;treated by traditional open reduction with plate internal fixation. Operation time, intraoperative blood loss, incision length, hospital stay, fracture healing time and lower limb full weight-bearing time and postoperative complications between two groups were observed and compared, bone mineral density, osteocalcin, blood calcium and phosphorus before operation and 1 month after operation.@*RESULTS@#All patients were followed up from 12 to 18 months with an average of (14.92±2.46) months. Operation time, intraoperative blood loss, incision length, hospital stay, fracture healing time and lower limb full weight-bearing time of external fixation group were significantly better than that of internal fixation group(@*CONCLUSION@#Locking plate external fixation combined with membrane induction technology in treating open and comminuted tibial fractures with severe post-traumatic bone defects has advantages of less trauma, reliable fixation, shorter fracture healing time, and could improve bone metabolic activity with less postoperative complications.

Comparative Study Between Periostin And Osteocalcin As Biomarkers For Osteoporosis And Fracture Risk In Egyptian Postmenopausal Women

Objective: This study aimed to compare between periostin and osteocalcin as biomarkers in Egyptian postmenopausal women with osteoporosis and to explore their possible relationship with fracture risk. Methods: This study included 90 postmenopausal females recruited from Al-Hussein University Hospital, Cairo, Egypt; divided into three groups; 35 postmenopausal osteoporotic females with low fracture risk (group I), 35 postmenopausal osteoporotic females with high fracture risk (group II), and 20 apparently healthy controls. Serum periostin, osteocalcin, and estrogen were measured by Enzyme Linked Immunosorbent Assay (ELISA). Fracture risk assessment was calculated. Alkaline phosphatase (ALP), total and ionized calcium, Aspartate transaminase (AST), and Alanine transaminase (ALT) were measured spectrophotometrically. Results: The diagnostic performance of periostin for discriminating high fracture risk from low fracture risk groups showed the specificity of (68.6 %) and sensitivity of (100 %), while for osteocalcin the specificity was (51.4 %) and the sensitivity was (68.6 %) respectively. Moreover, the multi Receiver Operating Characteristics (multi-ROC) curve for periostin and osteocalcin together revealed improved specificity and sensitivity of (100 %) each. Conclusion: Periostin was superior to osteocalcin in discriminating high fracture risk from low fracture risk postmenopausal osteoporotic groups. Moreover, dual use of both markers gave the highest discriminative power between low and high fracture risk groups with 100 % specificity and sensitivity.

Study of bone turnover markers, procollagen type 1 N-terminal propeptide and osteocalcin in healthy and type 2 diabetes mellitus postmenopausal women

Background: Individuals with diabetes mellitus are at increased risk of metabolic bone disease due to decrease in bone strength and quality. Several bone turnover markers like serum procollagen type I N propeptide (P1NP) and serum osteocalcin are powerful tools for studying osteoporosis and fracture risk across population to provide diagnostic and prognostic information of bone health. The aim of this study was to recognize possible correlation of levels of serum P1NP and osteocalcin in type-2 diabetic (T2DM) postmenopausal women as compared to healthy postmenopausal women.Methods: The study included 100 proven cases of type-2 diabetic postmenopausal women with age matched healthy postmenopausal women as controls. P1NP, osteocalcin, and other relevant parameters were measured. Differences between diabetics and controls were analyzed.Results: The body mass index was higher in diabetic group as compared to controls. The HbA1c% was (6.94±1.43) in diabetic group and (5.57±1.21) in non-diabetics. Low serum level of 25 (OH) D was observed both in diabetic and non-diabetic groups but significantly lower in T2DM. Procollagen type 1 N propeptide was lower in diabetic group (37.59±17.20 ng/mL) as compared to non-diabetic (52.14±24.82 ng/mL). Osteocalcin was lower (15.64±8.06 ng/ml) as compared to non-diabetic group (21.85±9.12 ng/ml). Lower osteocalcin and P1NP levels found in this study suggests slower bone metabolism with reduced bone formation in postmenopausal diabetics.Conclusions: Serum procollagen type 1 N propeptide and osteocalcin in postmenopausal diabetic women were lower as compared to non-diabetic group.

Study of bone formation marker levels in rheumatoid arthritis

Background: Rheumatoid arthritis (RA) is an autoimmune disease of unknown cause that affects the joints principally. The disease affects between 0.5% and 1% of the adult population worldwide. Two to three times as many women as men suffer from the disease. Osteocalcin (OC) is a small protein of 49 amino acids long. OC is the most abundant non-collagenous protein in bone. OC originates from osteoblasts and is deposited into bones or released into circulation, where it correlates with histological measures of bone formation. Bone alkaline phosphatase (ALP) is a glycoprotein that is found on the surface of osteoblasts. This enzyme reflects the biosynthetic activity of these bone-forming cells. The presence of OC and ALP in the circulation may, therefore, provide a specific chemical index of osteoblastic activity. Objectives: This study was undertaken to estimate the values of serum OC and ALP among patients with RA and healthy control groups and to compare and find out any changes in levels of serum OC and ALP between the study and control groups. Materials and Methods: It was a case–control study done on 76 RA patients and 76 age- and sex-matched healthy individuals. Serum OC and serum ALP values were evaluated among all 76 cases and 76 controls. Serum OC was measured using immunoenzymatric assay and ALP was measured by colorimetric method. Statistical analysis was performed and results were tabulated and analyzed. Results: Mean ± standard deviation of serum OC level is significantly higher (P < 0.001) among cases (18.50 ± 8.72 ng/ml) than controls (9.98 ± 7.68 ng/ml). Similarly, the values of ALP are higher (P < 0.001) among cases (216.22 ± 59.96 IU/L) than controls (164.17 ± 50.70 IU/L). A significantly positive correlation was found between serum OC and serum ALP levels. Patient with the highest mean value of serum OC also has the highest values of ALP. The values of serum ALP and OC levels increase significantly in both early and late stages when compared with control values. Conclusions: A significant difference between the values of serum OC and ALP among cases and controls was seen in the study. Levels of both these parameters are elevated in subjects with RA compared to controls. Furthermore, the levels of serum OC correlated with the levels of serum ALP. This study demonstrates that increased bone formation is associated with RA together with bone resorption.

Increased serum osteocalcin levels and vitamin K status by daily cheese intake

Background:Cheese is a major source of long-chained vitamin K2 variants. How intake of vitamin K2 rich cheese affects vitamin K and osteocalcin has not been studied. The aim was to establish a maximum efficacy dose (MED) after daily intake of vitamin K2-rich cheese (Jarlsberg®) based on increase in ratio between carboxylated and undercarboxylated osteocalcin during a five-week diet.Methods:20 healthy healthy volunteers (HV) were recruited. The daily intake of Jarlsberg®cheese in the study varied from 20 to 152g. Clinical investigation was performed initially and after three, four and five weeks with measurement of vital signs, hematological and biochemical variables, carboxylatedand undercarboxylated osteocalcin and vitamin K. The ratio OR=carboxylated/undercarboxylated osteocalcin was the main variable.Results:The MED decreased with treatment duration and was estimated to 57 g/day (95% CI: 47-67)after five weeks diet, resulting in a mean OR increase of 30% (95% CI: 23.8-36.8). Both OR and serum osteocalcin followed a quadratic dose response curve. For osteocalcin,a maximal increase of 46% was estimated at 59 g/day for five weeks. The serum content of long-chained vitamin K2 increased significantly with increasing cheese dose. The increase were mainly obtained the first three weeks and kept unchanged the following two weeks. The cheese doses close to the MED caused nearly significant reductionsin total cholesterol, LDL-cholesterol, the LDL/HDL ratio and significant reduction in the blood pressures after five weeks diet (p?0.05). Conclusions: MED of Jarlsberg® cheese was estimated to 57g/day. Daily intake of Jarlsberg®cheese increasedthe osteocalcin level, vitamin K2andpositively affected the lipid patterns and blood pressure.

Experimental acute sepsis reduced number of osteocalcin immunolabeled cells in periodontal ligament

Abstract The aim of this study was to evaluate the effect of acute sepsis in the periodontal ligament, alveolar and furcation bone in absence of periodontitis induction through histological and immunohistochemical analyses. A septic rat model was established by cecal ligation and puncture (CLP). Twelve rats were randomly divided into CLP (n=6) and Sham (n=6) groups. The animals were euthanized at 24 h and hemimandibles were submitted to histomorfometric (bone matrix, collagenous fibers, fibroblasts, osteocytes, inflammatory cells, and blood vessels) and immunohistochemical (BMP-2/4, RANKL and osteocalcin) evaluation in alveolar bone, furcation bone and periodontal ligament. Our results demonstrated that histomorphometric parameters were similar in alveolar bone, furcation bone and periodontal ligament of Sham and CLP rats. Regarding to immunohistochemical analyses, the number of BMP-2/4 and RANKL immunolabeled cells was also similar in both groups. Furthermore, it was detected a reduction in the osteocalcin immunolabeled cells in periodontal ligaments of CLP compared to Sham rats (p=0.0014). In conclusion, the acute sepsis induction resulted in reduced number of osteocalcin labelled cells in periodontal ligament region. Moreover, no significant histological differences were observed in the periodontium of rats under acute sepsis. Considering the role of osteocalcin in bone remodeling, the study contributes to revealing the importance of careful periodontal evaluation in the presence of sepsis.

Resumo O objetivo deste estudo foi avaliar o efeito da sepse aguda no ligamento periodontal, osso alveolar e osso da furca por meio de análise histológica e imunohistoquímica. O modelo de sepse em ratos foi estabelecido pelo procedimento de ligação e perfuração do ceco (CLP). Doze ratos foram divididos de forma randomizada em ratos sépticos (n=6) e controle - grupo Sham (n=6). Os animais foram eutanasiados após 24 horas e suas hemimandíbulas foram submetidas aos procedimentos histotécnicos para análise histomorfométricos (matriz óssea, fibras colágenas, fibroblastos, osteócitos, células inflamatórias e vasos sanguíneos) e imunohistoquímicos (BMP-2/4, RANKL e osteocalcina) no osso alveolar, osso de furca e ligamento periodontal. Nossos resultados demonstraram que os parâmetros histomorfométricos foram similares no osso alveolar, osso de furca e ligamento periodontal dos animais do grupo sepse e do grupo Sham. Em relação à análise por imunohistoquímica, o número de células imunomarcadas para BMP-2/4 e RANKL também foi similar em ambos os grupos. Entretanto, houve redução (p=0.0014) no número de células imunomarcadas para osteocalcina no ligamento periodontal de ratos sépticos em relação ao grupo Sham. Como conclusão, o estabelecimento de sepse aguda resultou em um número reduzido de células imunomarcadas para osteocalcina na região do ligamento periodontal (p=0,0014). Além disso, não foram observadas diferenças histológicas significativas no periodonto de ratos na presença de sepse aguda. Considerando o papel da osteocalcina na remodelação óssea, este estudo contribui para revelar a importância da avaliação periodontal cuidadosa na presença de sepse.

Study of Serum Osteocalcin Level as a Biomarker of Severity in Patients of NAFLD

Introduction: Osteocalcin has been considered an establishedmarker of bone formation; but the newer evidences suggestedits role in the obesity, metabolic syndrome, and type 2 diabetesmellitus. Thus, it plays an important part both in bone andenergy metabolism.Methodology: This study was conducted in the Department ofBiochemistry, Department of Pharmacology & Department ofMedicine, PMCH, Patna. Two groups were included in thisstudy. One is Non-NAFLD group and another group is NAFLD.Non-NAFLD group having 130 cases while NAFLD grouphaving 44 cases. The duration of study was over a period ofone year.Results: This study revealed that serum osteocalcin levels areinversely associated with NAFLD.Conclusion: This study conclude that, osteocalcin isconsidered as a biomarker of severity in patients of NAFLD.

Relationships between serum osteocalcin levels and C-peptide in post-traumatic male patients / 中华急诊医学杂志

Objective To investigate the relationships between serum osteocalcin (OC) levels and glycometabolism markers in nondiabetic post-traumatic male patients.Methods Populaitons were selected at the Department of Emergency Medicine of Shanghai Jiao Tong University Affiliated Sixth People's Hospital from October 2017 to February 2019.The age,injury severity score (ISS),and characteristic indicators were recorded.The inclusion criteria were age ≥ 18 years and blood collection time ＜ 24 h after the injury.The exclusion criteria were emergency surgery,acute brain trauma,and hemoglobin A1c (HbA1c) ≥ 6.0％.The patients were divided into two groups by fasting plasma glucose (FPG):stress hyperglycemia (SH) (FPG＞7.8 mmol/L) and nonstress hyperglycemia (NO-SH) (FPG ≤ 7.8 mmol/L) groups.The fasting venous blood samples were collected and examined.The characteristics and biochemical indicators in the two groups were compared statistically by LSD-t test,rank sum test and ANOVA,and the relationships between serum OC levels and glycometabolism markers were analyzed by partial correlation analysis.Results A total of 395 traumatic patients were enrolled and divided into the SH group (n=182) and NO-SH group (n=213).There were no differences in ISS,fasting insulin (FINS),and C-peptide (C-P) levels between groups.Age,HbAlc and FPG were higher (P=0.041,P=0.037,P＜0.01),while the OC level was lower (P=0.023),in the SH group than those in the NO-SH group.The serum OC level did not correlate with HbAlc,FPG,and FINS,but negatively correlated with C-P by partial correlation analysis (r=-0.262,P=0.008).The multivariate linear regression analysis showed that C-P was an independent factor affecting serum OC levels after trauma (β=-0.655,P=0.043).Conclusion A correlation existed between the serum OC level and glycometabolism markers in nondiabetic post-traumatic male patients.