HOXA13 upregulation in gastric cancer is associated with enhanced cancer cell invasion and epithelial-to-mesenchymal transition.

The article "HOXA13 upregulation in gastric cancer is associated with enhanced cancer cell invasion and epithelial-to-mesenchymal transition, by Y.-X. He, X.-H. Song, Z.-Y. Zhao, H. Zhao, published in Eur Rev Med Pharmacol Sci 2017; 21 (2): 258-265-PMID: 28165563" has been retracted by the authors due to controversial issues with author contributions. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/12092.

[Determination of hydrazine in workplace air by gas chromatography with large bore capillary column].

Objective: To determine hydrazine quantitatively in workplace air by gas chromatography with large bore capillary column. Methods: In October 2019, hydrazine in the air was adsorbed by acid silica gel tube sampling and desorped using sulfuric acid solution. After derivatization with furfural and extraction, the content of hydrazine was determined by DM-FFAP capillary column gas chromatography with flame ionization detector. Results: The linear regression equation was y=353.8x+21.2 (r=0.9998) between 0.1-2.0 µg/ml of target concentration. The detection limit was 0.030 µg/ml. The lower limit of quantification was 0.100 µg/ml. If 15 L air sample was collected, the minimum detection concentration was 0.004 mg/m(3) and the minimum quantitative concentration was 0.013 mg/m(3) respectively. The average desorption efficiency was 86.5%-89.4%. The recovery was 94.4%-97.1%. The relative standard deviation was 1.6%-4.9%. Hydrazine and furfural derivative was 2-furaldehyde hydrazine. Conclusion: The method has symmetrical peak shape of hydrazine derivatives chromatographic peaks, short analysis time, easy operation, and is suitable for the determination of the concentration of hydrazine in the air in the workplace.

MicroRNA-146a regulates the expression of the Aß1-42 protein in Alzheimer's disease.

OBJECTIVE: To investigate the expression and clinical significance of microRNA-146a, Aß1-42, and tau protein in the peripheral blood of patients with Alzheimer's disease (AD). PATIENTS AND METHODS: A total of 98 AD patients admitted to our hospital were selected as the experimental group and 50 healthy individuals were selected as the control group. The correlations between microRNA-146a, Aß1-42, and tau protein were analyzed using receiver operating curves to evaluate the value of microRNA-146a in predicting AD. Bioinformatics analysis was performed to preliminarily explore the possible mechanisms of microRNA-146a in the pathogenesis of AD. RESULTS: MicroRNA-146a, Aß1-42 and tau protein were differentially expressed between AD patients and healthy individuals (p<0.05). microRNA-146a was negatively correlated with Aß1-42 (r=-0.882, p<0.05) but was not correlated with tau protein (p>0.05). The ROC curve showed that the area under the curve for microRNA-146a could be used to predict AD with an accuracy of 0.879 (95% CI 0.812-0.947). Bioinformatics analysis showed that the differentially expressed genes (DEGs) of microRNA-146a may be involved in the pathogenesis of AD through the regulation of multiple signaling pathways, including the Toll-like receptor signaling pathway, the neurotransmitter regulatory signaling pathways, and other pathways that affect the inflammatory response, synapse formation, and other biological processes. CONCLUSIONS: MicroRNA-146a, Aß1-42 and tau protein are differentially expressed in the peripheral blood of AD patients. These proteins may be involved in the pathogenesis of AD by regulating the activity of multiple signaling pathways and the expression of the Aß1-42 protein.

Endovascular treatment of visceral aneurysms and pseudoaneurysms.

Visceral artery aneurysm (VAA) is a rare and potentially life-threatening condition, defined as true artery aneurysms and pseudoaneurysms of splanchnic circulation and renal artery. This study reports our experience in the diagnosis and endovascular treatment of visceral artery aneurysms (VAAs) over a 10-year period. Between 2008 and 2018, a total of 24 VAAs in 21 patients were diagnosed by clinical symptoms and a combination of imaging techniques, such as Doppler ultrasound, computed tomography angiogram, and catheter angiogram. All patients underwent endovascular treatment to exclude aneurysms. Oral antiplatelet medicine was administered, and imaging examination was performed during follow-up. Technical success was achieved in all 21 patients, and no periprocedural complications occurred. Endovascular coiling alone was employed in 10 aneurysms. Coiling was combined with gelfoam in 2 aneurysms. Coiling was assisted by stent in 4 aneurysms. Covered stents were deployed in 8 aneurysms individually. Clinical symptoms disappeared or highly improved in all patients after treatment. None of the patients showed recurrent symptoms after discharge. However, two cases with new aneurysms after 6 and 8 months, respectively, and one case with in-stent thrombosis after 12 months were reported during follow-up. This study may justify the efficacy of percutaneous endovascular coil embolization and stent deployment. It also provides beneficial experience about how to choose appropriate various endovascular strategies based on both clinical symptoms and aneurysm anatomy condition.

MiR-223 alleviates thrombus and inflammation in thromboangiitis obliterans rats by regulating NLRP3.

OBJECTIVE: The aim of this study was to observe the regulatory effects of micro ribonucleic acid (miR)-223 on thromboangiitis obliterans (TAO) rats, and to explore the potential regulatory mechanism. MATERIALS AND METHODS: Online database TargetScan was used to predict the downstream regulatory targets of miR-223. A total of 45 Sprague Dawley (SD) rats were randomly divided into three groups, including sham operation group (Sham group), Model group, and miR-223 agonist group (miR-223 mimic group). TAO model was successfully established in rats through the injection of lauric acid via the femoral artery. The content of serum thromboxane B2 (TXB2) and endothelin (ET) was measured via enzyme-linked immunosorbent assay (ELISA). The pathological changes in the left hind limb were detected via hematoxylin-eosin (HE) staining. Moreover, the expressions of interleukin-6 (IL-6) and IL-1ß in the tissues of the rat left hind limb were determined via immunohistochemistry. In addition, the protein expression of Nod-like receptor protein 3 (NLRP3) in tissues was determined using Western blotting. RESULTS: TargetScan database predicted that NLRP3 was the downstream target gene of miR-223. Compared with the Sham group, Model group exerted significantly higher content of serum TXB2 and ET, severe lesions in the rat left hind limb, as well as significantly increased expressions of IL-6 and IL-1ß and protein expression of NLRP3 in tissues of the rat left hind limb (p<0.05). Besides, compared with the Model group, miR-223 mimic group showed remarkably lower content of serum TXB2 and ET, improved lesions in the rat left hind limb, as well as decreased expressions of IL-6 and IL-1ß and protein expression of NLRP3 in the tissues of the rat left hind limb (p<0.05). CONCLUSIONS: MiR-223 agonist can alleviate thrombus and inflammatory response in TAO rats. The possible underlying mechanism may be related to targeted regulation on NLRP3 inflammasome expression.

[Long-term analysis of safety and efficacy of standard percutaneous nephrolithotomy in patients with solitary kidneys].

OBJECTIVE: To evaluate the long-term efficacy and safety of ultrasound-guided percutaneous nephrolithotomy (PCNL) in the treatment of patients with solitary kidney stones. METHODS: The clinical data of 22 patients with solitary kidney stones treated with PCNL in Peking University People's Hospital from September 2008 to June 2014, with the follow-up data of more than 5 years were analyzed retrospectively. Perioperative indicators, postoperative stone free rate (SFR) and incidence of complications were recorded. Ultrasonography was used to evaluate the long-term stones recurrence rate. Serum creatinine and estimated glomerular filtration rate (eGFR) were used to assess the long-term renal function. RESULTS: In this group of 22 patients, the average age was (50.3±11.8) years, with 10 cases of anatomic solitary kidneys, 12 functional solitary kidneys, and the median stone diameter was 1.65 (1.1-3.9) cm. All the patients had multiple stones, including 7 cases of staghorn stones. The median pre-operative serum creatinine was 104.5 (60.0-460.0) µmol/L, and the mean eGFR was (60.3±29.4) mL/min, showing no statistically significant difference compared with that before surgery. The mean operative time was (88.2±42.0) min, and there were 11 cases of single-channel and double-channel PCNL. The median serum creatinine on the first day after surgery was 102.0 (63.0-364.0) µmol/L, and the mean eGFR was (58.0±25.1) mL/min. The mean postoperative hospital stay was (8.7±5.2) days. In this group, 5 patients (22.7%) presented short-term complications, among which 4 patients presented postoperative infection and massive hemorrhage at the same time, which improved after conservative treatment, and 1 patient presented pleural injury and improved after closed thoracic drainage. Two patients (9.1%) developed long-term complications, and ureteral stricture occurred 3 months after operation, which improved after balloon dilatation. The median follow-up time was 6.2 (4.7-11.1) years. The median serum creatinine at the last follow-up was 104.0 (72.4-377.0) µmol/L, and the mean eGFR was (60.1±23.7) mL/min, showing no statistically significant difference compared with that before surgery. Renal function decreased in 6 patients (27.3%). Initial and final SFR were 72.7% and 100%, respectively. During the 6.2-year follow-up, 9 patients (40.9%) experienced recurrence of kidney stone. After stone recurrence, 13 lithotomy surgeries were performed, and the SFR by the latest follow-up was 63.6%. CONCLUSION: This study had the longest follow-up time for patients with solitary kidney stones after PCNL reported at home and abroad. Ultrasound-guided standard PCNL was safe and effective in the treatment of solitary kidney stones. Long-term follow-up results showed that the recurrence rate of kidney stones was still high, but the long-term renal function was stable after operation, and some patients showed mild renal function decline.

Up-regulation of exosomal miR-106a may play a significant role in abdominal aortic aneurysm by inducing vascular smooth muscle cell apoptosis and targeting TIMP-2, an inhibitor of metallopeptidases that suppresses extracellular matrix degradation.

OBJECTIVE: Abdominal aortic aneurysm (AAA) rupture is a dramatic and lethal clinical condition with a high risk of death. Emerging evidence indicates a role for miRNAs in AAA pathogenesis, therefore we aimed to identify miRNAs that differently expressed in exosomes from AAA patients and explore the underlying mechanisms of how miR-106a plays its role in this disease. PATIENTS AND METHODS: Exosomes were isolated from plasma of AAA patients, as well as from the tissue-conditioned culture medium. The exosomal expression profiles of several miRNAs including miR-106a were analyzed by quantitative RT-PCR. To determine the potential role of miR-106a in the pathogenesis of AAA, miR-106a was overexpressed in vascular smooth muscle cells (VSMCs), and then cell viability and apoptosis were evaluated by performing CCK-8 assay and flow cytometry, respectively. Afterwards, enzyme-linked immunosorbent assay (ELISA) was applied to assess the expression levels of some proteins involved in the modulation of extracellular matrix (ECM) homeostasis. Furthermore, the target gene of miR-106a was predicted and verified through Dual-Luciferase reporter assay. RESULTS: MiR-106a was up-regulated in exosomes from plasma of those patients with AAA as compared with healthy peers. Likely, increased level of miR-106a was observed in exosomes released from AAA tissue in comparison to those from adjacent normal tissues. Enhanced expression of miR-106a in VSMCs suppressed cell viability but promoted cellular apoptosis, whereas inhibition of miR-106a in VSMCs resulted in a significant decrease in the percentage of apoptotic cells compared to the control group. In addition, the protein levels of matrix metalloproteinases (MMPs, including MMP-2 and MMP-12) secreted from VSMCs were significantly up-regulated, while their inhibitor TIMP-2 was down-regulated due to miR-106a overexpression. Finally, TIMP-2 was validated subsequently as the direct target of miR-106a through Dual-Luciferase reporter assay. CONCLUSIONS: In aggregate, our results suggest that increased expression of miR-106a promotes VSMC cell apoptosis and down-regulates TIMP-2 through directly targeting its 3'-UTR, which in turn restores MMP production and ultimately accelerates ECM degradation. Therefore miR-106a is proposed to play a crucial role in AAA development and this will provide an update on the understanding of the clinical value of miRNAs as novel therapeutic targets for the treatment of this disease.

Association between hsa_circ_0006156 expression and incidence of gastric cancer.

OBJECTIVE: To screen the differentially expressed circular ribonucleic acids (circRNAs) related to gastric cancer and to explore their associations with the clinicopathological features of gastric cancer. PATIENTS AND METHODS: Cancer tissues of 50 gastric cancer patients undergoing surgical resection in our hospital from April 2015 to December 2018 were collected as an experimental group, while the para-carcinoma tissues were used as the control group. First, the differentially expressed circRNAs were screened by analyzing the circRNA profile in the microarray. Then, the expression of hsa_circ_0006156 in tissues was detected via Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) in both groups. The potential associations of the relative expression level of hsa_circ_0006156 with clinicopathological features and prognosis were analyzed according to the clinical data of gastric cancer patients. RESULTS: Six significantly downregulated circRNAs in gastric cancer patients were screened out. The results of RT-qPCR showed that the expression level of hsa_circ_0006156 was significantly lower in gastric cancer tissues than that in para-carcinoma tissues (p<0.05). Accordingly, 50 gastric cancer patients were divided into hsa_circ_0006156 high expression group and hsa_circ_0006156 low expression group based on the fold change of hsa_circ_0006156 in para-carcinoma tissues than that of gastric cancer tissues (fold change>3). The expression level of hsa_circ_0006156 was not correlated with the age and gender of gastric cancer patients (p>0.05) but correlated with the lymph node metastasis (p<0.05), nerve invasion (p<0.05), and degree of tumor differentiation (p<0.05). The expression level of hsa_circ_0006156 was also significantly associated with the progression-free survival (PFS) and overall survival (OS) of patients (p<0.05). According to the multivariate analysis of variance, the PFS of gastric cancer patients was associated with nerve invasion, lymph node metastasis, and hsa_circ_0006156 expression (relative risk coefficient=1.742, 2.329, and 3.003). Meanwhile, the OS was associated with lymph node metastasis, nerve invasion, degree of tumor differentiation, and hsa_circ_0006156 expression (relative risk coefficient =1.604, 2.405, 2.114, and 2.004). Moreover, the survival analysis revealed that PFS was markedly prolonged in the hsa_circ_0006156 high expression group compared with that in the hsa_circ_0006156 low expression group. CONCLUSIONS: The expression of hsa_circ_0006156 substantially declines in gastric cancer tissues, which is related to the differentiation degree, presence, or absence of lymph node metastasis and prognosis of gastric cancer patients. Therefore, hsa_circ_0006156 may clinically serve as a biomarker for the prognostic prediction of gastric cancer patients.

[Long-term follow-up of the efficacy of tension-free vaginal tape and trans-obturator tape for different types of stress urinary incontinence].

OBJECTIVE: To evaluate the long-term efficacy of tension-free vaginal tape (TVT)and trans-obturator tape (TOT)for different types of female stress urinary incontinence (FSUI). METHODS: The clinical data of all female patients with stress urinary incontinence (SUI) who underwent mid-urethral slings (MUS) in Peking University People's Hospital from January 2008 to June 2016 were retrospectively analyzed, and all the patients were followed up. Based on the level of abdominal leak point pressure (ALPP), the patients with ALPP lower than or equal to 60 cmH2O (1 cmH2O=0.74 mmHg) were regarded as intrinsic sphincter deficiency (ISD) type, and the patients with ALPP higher than 60 cmH2O were regarded as non-ISD type. According to the degree of remission of leakage of urine by using the International Consultation on Incontinence Questionnaire Short Form (ICI-Q-SF), the surgical outcomes were divided into three categories: cured, improved or failed. The efficacy of TVT and TOT for different types of FSUI was evaluated by comparing the cure rate of TVT with the cure rate of TOT in ISD type and non-ISD type separately. RESULTS: A total of 170 patients were included in this study. The patients were among 30-78 years, and the follow-up period were among 12-110 months. In the study, 117 patients belonged to non-ISD type. Of whom 30 underwent TVT with a cure rate of 86.7% (26/30), and the other 87 underwent TOT with a cure rate of 69.0% (60/87). The cure rate of TVT was higher than that of TOT in non-ISD type, however, Chi-square test showed that there was no significant difference (χ2=3.589, P>0.05). In addition, 53 patients belonged to ISD type, of whom 16 underwent TVT with a cure rate of 87.5% (14/16), and the other 37 underwent TOT with a cure rate of 51.4% (19/37). The cure rate of TVT was higher than that of TOT in ISD type, and Chi-square test showed that the difference was statistically significant (χ2=6.212, P<0.05). CONCLUSION: For the treatment of FSUI, MUS can achieve satisfactory long-term efficacy. For non-ISD type, the cure rates of TVT and TOT are similar, while for ISD type, the cure rate of TVT is higher than that of TOT.

Vessel traffic safety in busy waterways: A case study of accidents in western shenzhen port.

Throughout the world, busy waterways near large ports witness heavy vessel traffic in recent decades. The waterways are characterized by high risk in terms of loss of life, property, and pollution to environment. To facilitate maritime safety management with satisfactory efficiency and efficacy, the authors propose a framework of safety indexes to evaluate the risk level in busy waterways according to the accident severity, fatality rate and special indicators of maritime transportation. The safety indexes consist of Safety Evaluation Index (SEI) and Safety Warning Index (SWI), and are derived from the proposed risk criteria of Chinese vessel traffic. As a case study, data on vessel traffic accidents reported in the Western Shenzhen Port, South China from 1995 to 2015 are analyzed. The actual risk level of this area during the period is calculated under the framework. The implementation of the safety indexes indicate that the risk criteria and safety indexes are practicable and effective for the vessel traffic management. The methodology based on long-term accident data can significantly support the risk analysis in the macroscopic perspective for busy ports and waterways, such that SWI can act as threshold to trigger actions, while SEI can act as an indicator to measure safety status.

[Simultaneous determination of 21 organophosphrous pesticides in human serum by QuEChERS-gas chromatography-mass spectrometry].

Objective: A method for the simultaneous determination of organophosphrous pesticides inhuman serum was developed. Methods: Using C18 and PSA as absorbent, 21 organophosphrous pesticides including methamidophos, dichlorvos, acephate, Omethoate, demeton and etc. in human serum were determined by combing QuEChERS with GC-MS, organophosphrous pesticides were qualitatedsimultaneously by retention time and characteristic ions. Their contents were determined by internal standard method of quantitative ions. Results: The linear range was from0to 0.60µg/ml with a good correlation coefficients (r ≥ 0.9961) . The LODs of this methodranged from 0.0009 to 0.0111 µg/ml. The average recoveries of3 organophosphrous pesticides (methamidophos、dichlorvos and acephate) ranged from 71.8%~85.3%, The average recoveries of other 18 organophosphrous pesticidesranged from 82.9%~102.9%. The relative standard deviations were0.8%~7.0%. Conclusion: The convenient operation and versatility of this method are suitable forthe fast screening and accurate detection of 21 organophosphrous pesticides in human serum.

HOXA13 upregulation in gastric cancer is associated with enhanced cancer cell invasion and epithelial-to-mesenchymal transition.

OBJECTIVE: In this study, we investigated the association between HOXA13 dysregulation and gastric cancer progression. We also explored the functional role of HOXA13 in invasion and epithelial-to-mesenchymal transition (EMT) of gastric cancer cells and the possible signaling pathway it might involve in. MATERIALS AND METHODS: The microarray (E-GEOD-19826) examined the transcription profiles of 12 adjacent normal/tumor-matched gastric tissues was downloaded from the ArrayExpress and reanalyzed. Immunohistochemistry (IHC) staining was performed to assess HOXA13 expression in 23 stage I and 69 stage II/III/IV gastric cancer tissues. The human gastric cancer cell line AGS and SGC-7901 cells were transfected with HOXA13 siRNA and then were subjected to detection of epithelial and mesenchymal markers and cell invasion. The involvement of HOXA13 in TGF-ß signaling was further studied. RESULTS: HOXA13 is one of the most upregulated genes in gastric cancer tissues compared to adjacent normal tissues. Also, HOXA13 is further upregulated in the higher stage tumors. HOXA13 staining was significantly stronger in stage II/III/IV tumors than in stage I tumors. HOXA13 siRNA significantly restored the epithelial property and reduced the mesenchymal property of the cancer cells. Transwell assay showed that HOXA13 siRNA impaired the invasion capability of the cancer cells. The gastric cancer cells with HOXA13 knockdown had decreased expression of p-SMAD2 and p-SMAD3. CONCLUSIONS: This study provides additional evidence about the association between HOXA13 upregulation and gastric cancer progression. Also, we showed that HOXA13 contributes to invasion and EMT of gastric cancer cells via the TGF-b signaling pathway.

[The role definition of lateral arm free flap in reconstruction after head and neck cancer surgery].

Application of free flap is one of the important repair means in head and neck surgery. A variety of free flaps, such as anterolateral thigh flap, have showed unique advantages in repair for tissue defects after resection of head and neck tumor, and have became increasing popularity. Lateral arm flee flap is an important repair means in plastic surgery, which has developed more than 30 years, but the application of this flap for reconstruction in head and neck surgery is relatively backward, with few reports. This review focuses on the creativity and innovation, the relationship between anatomy and clinical application, and the application status and prospects for lateral arm flee flap in individual head and neck reconstruction surgery.

A targeting drug delivery system for ovarian carcinoma: transferrin modified lipid coated paclitaxel-loaded nanoparticles.

The transferring modified lipid coated PLGA nanoparticles, as a targetable vector, were developed for the targeting delivery of anticancer drugs with paclitaxel (PTX) as a model drug to the ovarian carcinoma, which combines the advantages and avoids disadvantages of polymeric nanoparticles and liposomes in drug delivery. A transmission electron microscopy (TEM) confirmed the lipid coating on the polymeric core. Physicochemical characterizations of TFLPs, such as particle size, zeta potential, morphology, encapsulation efficiency, and in vitro PTX release, were also evaluated. In the cellular uptake study, the TFLPs were more efficiently endocytosed by the A2780 cells with high expression of transferrin receptors than HUVEC cells without the transferrin receptors. Furthermore, the anticancer efficacy of TFLPs on the tumor spheroids was stronger than that of lipid coated PLGA nanoparticles (LPs) and PLGA nanoparticles. In the in vivo study, the TFLPs showed the best inhibition effect of the tumor growth for the ovarian carcinoma-bearing mice. In brief, the TFLPs were proved to be an efficient targeting drug delivery system for ovarian carcinoma.

The evaluation of Lugol's iodine solution staining combined with endoscope for diagnosis of non-erosive reflux disease in clinic.

AIM: To evaluate the value of Lugol's iodine solution staining combined with endoscope on the diagnosis of non-erosive reflux disease (NERD). PATIENTS AND METHODS: A total of 96 gastroesophageal reflux disease patients were selected to participate in this study. The patients were stained on esophageal mucosa by Lugol's iodine solution and examined at routine endoscopy. The shallow staining and/or non-staining group patients were treated with esomeprazole and mosapride citrate, and then the changes in Lugol's iodine staining, Gerd Q (Gerd questionnaire) scoring and histological characters of esophageal mucosa were recorded before and after treatment. RESULTS: As the results, a total of 68 patients were diagnosed as NERD, and 36 of 68 patients were observed with uniform staining and 32 of 68 patients were observed with shallow staining and/or non-staining. After 4 weeks for treatment, 28 of 32 patients with shallow staining and/or non-staining became uniform staining and 4 of 32 patients were still with shallow staining and/or non-staining. Before and after treatment, the Gerd Q scoring of uniform staining groups and shallow staining and/or non-staining groups all had a significant difference (p < 0.05). Compared to the routine endoscopy, the detection rate of abnormal esophageal mucosa was significantly increased by Lugol's iodine solution staining combined with endoscope. CONCLUSIONS: Lugol's iodine solution staining combined with routine endoscopy, Gerd Q scoring and histomorphology can be used to evaluate the diagnosis and therapeutic effect of NERD.

Deletion of estrogen receptor beta accelerates early stage of bone healing in a mouse osteotomy model.

UNLABELLED: This study examined the role of estrogen receptor (ER) beta during mouse femoral fracture healing by employing ER knockout (KO) mice. The fracture healing in KO mice was enhanced in the early stage of neovascularization and the middle stage of endochondral ossification. INTRODUCTION: This study was conducted to examine the role of ER beta during fracture healing. METHODS: Female ERbeta knockout (KO) mice (18 weeks old) and age-matched female wild-type (WT) mice underwent open osteotomy on the right femur. They were sacrificed at 1, 2, 4 and 6 weeks post-fracture. The sera and callus samples were subjected to the following analyses: micro-computed tomography (CT)-based angiography, micro-CT evaluation, histological examination, histomorphometry examination, real-time polymerase chain reaction (PCR) analysis, biochemical marker, and mechanical testing. RESULTS: Micro-CT-based angiography showed that the total vessel volume at the fracture site was larger in the KO group than the WT group at 1 and 2 weeks post-fracture. Micro-CT analysis revealed that the callus volume was significantly higher in the KO group from week 2 to week 4 post-fracture when compared with the WT group consistent with the histological data. Analysis of biochemical markers indicated that circulating P1NP levels in the KO mice were significantly higher than in the WT mice from week 2 to week 4 and that temporal expression of circulating C-terminal telopeptide of type I collagen (CTX) levels was also higher in the KO mice than in the WT mice. These results were consistent with quantitative real-time PCR analysis. The ultimate load, stiffness, and energy to failure were significantly higher in the KO mice than in the WT mice at week 4. CONCLUSIONS: The fracture healing in KO mice was enhanced in the early stage of neovascularization and the middle stage of endochondral ossification, but not by the end of healing. Blockade of ERbeta can be considered as another therapeutic strategy for osteoporotic fracture and non-union fracture.