Annexin A5 derived from matrix vesicles protects against osteoporotic bone loss via mineralization.

Matrix vesicles (MVs) have shown strong effects in diseases such as vascular ectopic calcification and pathological calcified osteoarthritis and in wound repair of the skeletal system due to their membranous vesicle characteristics and abundant calcium and phosphorus content. However, the role of MVs in the progression of osteoporosis is poorly understood. Here, we report that annexin A5, an important component of the matrix vesicle membrane, plays a vital role in bone matrix homeostasis in the deterioration of osteoporosis. We first identified annexin A5 from adherent MVs but not dissociative MVs of osteoblasts and found that it could be sharply decreased in the bone matrix during the occurrence of osteoporosis based on ovariectomized mice. We then confirmed its potential in mediating the mineralization of the precursor osteoblast lineage via its initial binding with collagen type I to achieve MV adhesion and the subsequent activation of cellular autophagy. Finally, we proved its protective role in resisting bone loss by applying it to osteoporotic mice. Taken together, these data revealed the importance of annexin A5, originating from adherent MVs of osteoblasts, in bone matrix remodeling of osteoporosis and provided a new strategy for the treatment and intervention of bone loss.

AnnexinA6: a potential therapeutic target gene for extracellular matrix mineralization.

The mineralization of the extracellular matrix (ECM) is an essential and crucial process for physiological bone formation and pathological calcification. The abnormal function of ECM mineralization contributes to the worldwide risk of developing mineralization-related diseases; for instance, vascular calcification is attributed to the hyperfunction of ECM mineralization, while osteoporosis is due to hypofunction. AnnexinA6 (AnxA6), a Ca2+-dependent phospholipid-binding protein, has been extensively reported as an essential target in mineralization-related diseases such as osteoporosis, osteoarthritis, atherosclerosis, osteosarcoma, and calcific aortic valve disease. To date, AnxA6, as the largest member of the Annexin family, has attracted much attention due to its significant contribution to matrix vesicles (MVs) production and release, MVs-ECM interaction, cytoplasmic Ca2+ influx, and maturation of hydroxyapatite, making it an essential target in ECM mineralization. In this review, we outlined the recent advancements in the role of AnxA6 in mineralization-related diseases and the potential mechanisms of AnxA6 under normal and mineralization-related pathological conditions. AnxA6 could promote ECM mineralization for bone regeneration in the manner described previously. Therefore, AnxA6 may be a potential osteogenic target for ECM mineralization.

Fluid Shear Stress Regulates Osteogenic Differentiation via AnnexinA6-Mediated Autophagy in MC3T3-E1 Cells.

Fluid shear stress (FSS) facilitates bone remodeling by regulating osteogenic differentiation, and extracellular matrix maturation and mineralization. However, the underlying molecular mechanisms of how mechanical stimuli from FSS are converted into osteogenesis remain largely unexplored. Here, we exposed MC3T3-E1 cells to FSS with different intensities (1 h FSS with 0, 5, 10, and 20 dyn/cm2 intensities) and treatment durations (10 dyn/cm2 FSS with 0, 0.5, 1, 2 and 4 h treatment). The results demonstrate that the 1 h of 10 dyn/cm2 FSS treatment greatly upregulated the expression of osteogenic markers (Runx2, ALP, Col I), accompanied by AnxA6 activation. The genetic ablation of AnxA6 suppressed the autophagic process, demonstrating lowered autophagy markers (Beclin1, ATG5, ATG7, LC3) and decreased autophagosome formation, and strongly reduced osteogenic differentiation induced by FSS. Furthermore, the addition of autophagic activator rapamycin to AnxA6 knockdown cells stimulated autophagy process, and coincided with more expressions of osteogenic proteins ALP and Col I under both static and FSS conditions. In conclusion, the findings in this study reveal a hitherto unidentified relationship between FSS-induced osteogenic differentiation and autophagy, and point to AnxA6 as a key mediator of autophagy in response to FSS, which may provide a new target for the treatment of osteoporosis and other diseases.

Vascular Calcification: New Insights Into BMP Type I Receptor A.

Vascular calcification (VC) is a complex ectopic calcification process and an important indicator of increased risk for diabetes, atherosclerosis, chronic kidney disease, and other diseases. Therefore, clarifying the pathogenesis of VC is of great clinical significance. Numerous studies have shown that the onset and progression of VC are similar to bone formation. Members of the bone morphogenetic protein (BMP) family of proteins are considered key molecules in the progression of vascular calcification. BMP type I receptor A (BMPR1A) is a key receptor of BMP factors acting on the cell membrane, is widely expressed in various tissues and cells, and is an important "portal" for BMP to enter cells and exert their biological effect. In recent years, many discoveries have been made regarding the occurrence and treatment of ectopic ossification-related diseases involving BMP signaling targets. Studies have confirmed that BMPR1A is involved in osteogenic differentiation and that its high expression in vascular endothelial cells and smooth muscle cells can lead to vascular calcification. This article reviews the role of BMPR1A in vascular calcification and the possible underlying molecular mechanisms to provide clues for the clinical treatment of such diseases.

Fluid shear stress activates YAP to promote epithelial-mesenchymal transition in hepatocellular carcinoma.

Epithelial-mesenchymal transition (EMT) mediated by fluid shear stress (FSS) in the tumor microenvironment plays an important role in driving metastasis of the malignant tumor. As a mechanotransducer, Yes-associated protein (YAP) is known to translocate into the nucleus to initiate transcription of genes involved in cell proliferation upon extracellular biophysical stimuli. Here, we showed that FSS facilitated cytoskeleton rearrangement in hepatocellular carcinoma cells, which led to the release of YAP from its binding partner, integrin ß subunit, in the cytomembrane. Moreover, we found that upregulation of guanine nucleotide exchange factor (GEF)-H1, a microtubule-associated Rho GEF, is a critical step in the FSS-induced translocation of YAP. Nuclear YAP activated the expression of the EMT-regulating transcription factor SNAI1, but suppressed the expression of N6-methyladenosine (m6 A) modulators; together, this promoted the expression of EMT-related genes. We also observed that FSS-treated HepG2 cells showed markedly increased tumorigenesis and metastasis in vivo. Collectively, our findings unravel the underlying molecular processes by which FSS induces translocation of YAP from the cytomembrane to the nucleus, contributes to EMT and enhances metastasis in hepatocellular carcinoma.

Autophagy modulates FSS-induced epithelial-mesenchymal transition in hepatocellular carcinoma cells.

Hepatocellular carcinoma is a highly fatal disease and threatens human health seriously. Fluid shear stress (FSS), which is caused by the leakage of plasma from abnormally permeable tumor blood vessels and insufficient lymphatic drainage, has been identified as contributing pathologically to cancer metastasis. Autophagy and epithelial-mesenchymal transition (EMT) are both reported to be involved in cancer cell migration and invasion, but little has been revealed about the interaction between autophagy and EMT under a tumor mechanical microenvironment. Here, we identified that exposure to 1.4 dyne/cm2 FSS could promote the formation of autophagosomes and significantly increase the expressions of autophagy-related markers of beclin1 and ATG7, and the ratio of LC3Ⅱ/Ⅰ in both of HepG2 and QGY-7703 cells. The FSS loading also elevated the levels of mesenchymal markers N-cadherin, Vimentin, Twist, Snail, and ß-catenin, while the epithelial markers E-cadherin showed a decrease. Once the autophagy was blocked by 3-methyladenine (3-MA) or knocking ATG5 down, the occurrence of FSS-induced EMT was inhibited dramatically according to the expression and translocation of E-cadherin, N-cadherin, and ß-catenin. Given the effect of EMT on cell migration, we observed that inhibition of autophagy could impede FSS-induced cell migration. Collectively, this study demonstrated that autophagy played a crucial role in FSS-induced EMT and cell migration in hepatocellular carcinoma.

Identifying competing interest disclosures in systematic reviews of surgical interventions and devices: a cross-sectional survey.

BACKGROUND: A competing interest is an important source of bias in research and disclosure is frequently employed as a strategy to manage it. Considering the importance of systematic reviews (SRs) and the varying prevalence of competing interests in different research fields, we conducted a survey to identify the range of competing interests in SRs assessing surgical interventions or devices and explored the association between the competing interest disclosures and authors' conclusions. METHODS: We retrieved SRs of surgical interventions and devices published in 2017 via PubMed. Information regarding general characteristics, funding sources, and competing interest disclosures were extracted. We conducted a descriptive analysis of the studies' characteristics and compared them between Cochrane SRs (CSRs) and non-Cochrane SRs using the Chi-square test. Results were expressed as odds ratio and their 95% confidence interval. RESULTS: One hundred fifty-five SRs published in 2017 were included in the study. More than half of the SRs (58.7%) reported their funding sources and 94.2% reported authors' competing interest disclosures. Among 146 SRs that stated competing interest disclosures, only 35 (22.6%) SRs declared at least one author had a competing interest. More than 40 terms were used to describe competing interests. Cochrane SRs (CSRs) were more likely to provide a detailed description of competing interests compared to those in non-CSRs (48.0% versus 25.4%, P = 0.023). No association between positive conclusions and competing interest disclosures was found (P = 0.484, OR = 0.43, 95%CI: 0.08, 2.16). In the subgroup analyses, SRs stating no competing interest disclosure were more likely to report positive conclusions than those stating at least one type of competing interest, but the difference is not significantly different (P = 0.406, OR = 1.38, 95%CI: 0.64, 2.98). CONCLUSION: In surgical SRs, there is a high percentage of competing interest disclosures but without detailed information. The identification and statement of competing interests with a detailed description, particularly the non-financial ones, needs improvement. Some efficient and effective methods/tools for identifying, quantifying, and minimizing potential competing interests in systematic reviews remains valuable.

[Effect of conditioned medium of vascular endothelial cells on the epithelial-mesenchymal transition of hepatocellular carcinoma cells].

This study aims to investigate the effect of substances secreted or metabolized by vascular endothelial cells on epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma cells under indirect co-culture condition. Human hepatocellular carcinoma cell line QGY-7703 was cultured in vitro, and then was co-cultured with conditioned medium of human umbilical vein endothelial cells (HUVEC). The morphological changes of QGY-7703 cells were observed by inverted phase contrast microscopy. The migration ability of QGY-7703 cells was analyzed by scratch-wound assays. The effect of conditioned medium on the expression and distribution of EMT related proteins was detected by Western blot and immunofluorescence assays, respectively. The results showed that the QGY-7703 cells gradually changed from polygonal to spindle shape, the migration ability promoted significantly, and both the expression and distribution of EMT related marker changed in a time-dependent manner after co-culturing. The results confirm that vascular endothelial cells can induce EMT in hepatocellular carcinoma cells under indirect co-culture condition.

[Effects of arsenic trioxide on migration, invasion and apoptosis of hepatocellular carcinoma HepG2 cells].

The article aims to explore the optimal concentration of arsenic trioxide (As 2O 3) on HepG2 of liver cancer cells, and the effect of As 2O 3 on the migration, invasion and apoptosis of HepG2 cells. In this study, the activity of HepG2 cells treated with 0, 1, 2, 4, 8, 16, 32 µmol/L As 2O 3 was tested by CCK-8 method, the semi-inhibitory concentration (IC50) was calculated, and the morphological changes of HepG2 cells were observed after the action of As 2O 3 at IC50 concentration for 12, 24, 48 h. The effect of As 2O 3 on cell migration and invasion ability was verified by wound healing experiment and Transwell invasion experiment. Western blot and qRT-PCR were used to detect the effects of As 2O 3 on the gene and protein expression levels related to cell migration, invasion and apoptosis. The results showed that, compared with the control group, the activity of HepG2 cells decreased with the increase of the concentration of As 2O 3 treatment, showing a dose-dependent effect, and its IC50 was 7.3 µmol/L. After 24 hours' treatment with 8 µmol/L As 2O 3, HepG2 cells underwent significant apoptosis, and its migration and invasion abilities were significantly reduced. In addition, the protein expression levels of RhoA, Cdc42, Rac1 and matrix metalloproteinase-9 (MMP-9) were down-regulated, the protein and mRNA expression levels of anti-apoptotic gene Bcl-2 were significantly down-regulated, and the protein and mRNA expression levels of pro-apoptotic genes Bax and Caspase-3 were significantly up-regulated. The above results indicate that certain concentration of As 2O 3 can inhibit the migration and invasion of hepatocellular carcinoma cells and promote the apoptosis of hepatocellular carcinoma cells.

Motherwort Injection for Preventing Postpartum Hemorrhage in Women with Vaginal Delivery: A Systematic Review and Meta-Analysis of Randomized Evidence.

BACKGROUND: Motherwort injection, a common traditional Chinese medicine, is widely used for the prevention of postpartum hemorrhage (PPH), which has been found to be potential benefit in clinical practice. OBJECTIVES: This study aimed to conduct a rigorous systematic review of randomized evidence to offer a comprehensive overview regarding the efficacy and safety of motherwort injection in maternal women with virginal delivery. METHODS: We included all randomized controlled trials involving pregnant women in vaginal delivery comparing motherwort injection or combination of motherwort injection and oxytocin with oxytocin alone for preventing postpartum hemorrhage. Paired reviewers independently screened citations, assessed risk of bias, and extracted data. Random-effects model by Mantel-Haenszal method was applied to pool the data. Predefined subgroup analyses and sensitivity analyses were conducted to explore the heterogeneity and robustness of results. The GRADE approach was used to rate the quality of evidence. MAIN RESULTS: 37 randomized controlled trials involving 7887 participants were included, all of which were at moderate to high risk of bias. Meta-analyses of eight trials showed no significant difference in blood loss and PPH events between oxytocin versus motherwort injection (very low quality). However, pooling of 29 trials suggested a reduced risk of blood loss (within 2 hours: MD -55.06mL, 95% CI -84.06 to -26.06; within 24 hours: MD -85.57 mL, 95% CI -94.26 to -76.88, very low quality), PPH events (RR 0.29, 95% CI 0.21 to 0.39, low quality), and adverse events (Peto OR 0.53, 95% CI 0.40 to 0.70, very low quality) in participants treated with motherwort injection and oxytocin versus oxytocin alone. CONCLUSIONS: The current evidence supports the suggestion that the additional use of motherwort injection on oxytocin had a preferable outcome. However, given that the evidence is not definitive with low quality, further careful designed and conducted randomized controlled trials in larger population are warranted to conform the effects.

Causal Effects of Genetically Predicted Cardiovascular Risk Factors on Chronic Kidney Disease: A Two-Sample Mendelian Randomization Study.

Observational studies have demonstrated that cardiovascular risk factors are associated with chronic kidney disease (CKD). However, these observational associations are potentially influenced by the residual confounding, including some unmeasured lifestyle factors and interaction risk factors. Two-sample mendelian randomization analysis was conducted in this study to evaluate whether genetically predicted cardiovascular risk factors have a causal effect on the risk of CKD. We selected genetic variants associated with cardiovascular risk factors and extracted the corresponding effect sizes from the largest GWAS summary-level dataset of CKD. Cardiovascular risk factors contain high density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol, total cholesterol (TC), triglyceride (TG), glycated hemoglobin (HbA1c), fasting glucose, systolic blood pressure (SBP) and diastolic blood pressure (DBP). A Bonferroni corrected threshold of P = 0.006 was considered as significant, and 0.006 < P < 0.05 was considered suggestive of evidence for a potential association. Genetically predicted DBP was significantly associated with CKD [odds ratio (OR) was 1.35 (95% confidence interval (CI) (1.10, 1.65); P = 0.004)]. There was suggestive evidence for potential associations between genetically predicted higher HDL cholesterol [OR: 0.88, 95%CI (0.80, 0.98), P = 0.025] and lower adds of CKD, and between higher SBP [OR: 1.36, 95%CI (1.07, 1.73), P = 0.013] and higher adds of CKD. However, genetically predicted LDL cholesterol, TC, TG, HbA1c, and fasting glucose did not show any causal association with CKD.

Fluid shear stress induces cell migration and invasion via activating autophagy in HepG2 cells.

Fluid shear stress (FSS) regulates the metastasis of hepatocellular carcinoma (HCC). In the present study, we aimed to study the role of autophagy in HCC cells under FSS. The results showed that FSS upregulated the protein markers of autophagy, induced LC3B aggregation and formation of autophagosomes. Inhibition of integrin by Cliengitide (Cli) or inhibition of the microfilaments formation both inhibited the activation of autophagy in HepG2 under FSS. In addition, Cli inhibited the microfilaments formation and expressions of Rac1 and RhoA in HepG2 cells under FSS. Finally, inhibition of autophagy suppressed the cell migration and invasion in HepG2 under FSS. In conclusion, FSS induced autophagy to promote migration and invasion of HepG2 cells via integrin/cytoskeleton pathways.

Quality of reporting in randomized controlled trials of therapeutic cardiovascular medical devices.

BACKGROUND: Therapeutic medical devices play an important role in the treatment of cardiovascular diseases. The reliability of the randomized controlled trial, which is the best design for assessing treatment effects, largely depends on the information found in published reports. Limited information regarding the quality of reporting about therapeutic medical devices in trials was provided. METHOD: A cross-sectional study was conducted to assess the reporting quality of randomized controlled trials that tested the effects of therapeutic cardiovascular medical devices. The quality of reporting was assessed against a modified Consolidated Standards of Reporting Trials checklist, including 47 items from the Consolidated Standards of Reporting Trials statement and Consolidated Standards of Reporting Trials extension. We also examined the specific items regarding medical devices. Univariable and multivariable linear regressions were undertaken to explore potential factors associated with Consolidated Standards of Reporting Trials scores. RESULT: Some 115 randomized controlled trials were identified. The mean (standard deviation) Consolidated Standards of Reporting Trials score was 20.5 (5.0). The extent of compliance with the Consolidated Standards of Reporting Trials reporting guideline differed substantially across items: 5 of the 47 items were reported adequately across trials (more than 90%), and 10 were reported adequately in less than 5% of trials. Less than 50% of the trials reported additional items related to the medical device. Multivariable regression analysis showed that trials published in general journals (coefficient 7.44, 95% confidence interval [CI]: 5.50-9.38), with larger sample sizes (coefficient 2.30, 95% CI: 0.76-3.83), and multiple-center studies (coefficient 3.14, 95% CI: 1.27-5.01) were associated with a higher quality of reporting. CONCLUSION: The overall reporting quality in randomized controlled trials of therapeutic medical device trials is suboptimal, particularly in terms of items regarding surgeons and hospitals. We suggest that the existing Consolidated Standards of Reporting Trials and extension should be modified to be more applicable to therapeutic medical devices.

Integrin-Induced Signal Event Contributes to Self-Assembled Monolayers on Au-Nanoparticle-Regulated Cancer Cell Migration and Invasion.

Gold nanoparticles (Au NPs) have received much attention because of their distinct physicochemical properties. The surface terminal functional groups of Au NPs can facilitate easy conjugation with biological molecules for targeting cancer cells and controlling drugs/genes release. However, little is known regarding molecular mechanisms involved in their regulation of cancer cell migration and invasion. In the present study, Au NPs were successfully conjugated with functional groups (CH3, NH2, OH and COOH) by self-assembled monolayer (SAM) technique. The endocytosis of SAM-Au NPs mediating HepG2 cell migration and invasion in integrin-induced cascaded events were examined. Our results showed that the combination of integrins-Caveolin-1 together contributed to the internalization of SAM-Au NPs. The CH3-Au NPs showed fast cell motility than COOH- and OH- groups by upregulating PI3K expression, but reducing FAK phosphorylation level. Additionally, CH3-Au NPs showed the strongest activated GTP-bound Rac1 and RhoA. Taken together, these results concluded that internalization of SAM-Au NPs inhibited cancer cell migration via FAK/PI3K and downstream Rho-GTPase signaling pathway in a time-dependent manner. This work provides a further understanding of SAM-Au NPs regulating cancer cell migration, which might be helpful to functionalize the Au NP surface in drug delivery system.

Design, Conduct, and Analysis of Surgical Randomized Controlled Trials: A Cross-sectional Survey.

BACKGROUND: Randomized controlled trial (RCT) testing surgical intervention faced challenges due to complexities of surgery and made it more difficult for surgeons and methodologists than pharmaceutical providers to build a well-design, conduct RCT. OBJECTIVE: We conducted a cross-sectional survey to address the methodological challenges of RCTs on surgical intervention and offer potential solutions. METHODS: We searched PubMed in order to summarize 2-arm parallel randomized trials for surgical interventions published in 2013. The information regarding general characteristics, general methodological and special surgical characteristics related to surgical trials comparing alternative procedures was gathered. RESULTS: Some 200 surgical trials were identified. The extent to which these trials in design, conduct and analysis differed substantially across items. The general information about sample size calculation (77.0%), lost to follow-up (71.5%), trial registration (55.5%), protocols of trials (56.0%), implementation of randomization (59.5%), concealment of randomization (56.0%); reporting of primary outcome as P value (67.0%). Surgery special information revealed that only 21.0% of trials considered surgeons' preference, approximately 12% to 50% of them controlled the quality of surgical interventions and none evaluated the effect of the learning curve. CONCLUSION: There is much room for improvement concerning the reported designs, conduct, and analysis of surgical RCTs. Considering the difficulty of surgical RCTs, some other approaches, such as surgeons' eligibility, performance of pilot studies, or implementation of pragmatic RCTs/expertise-based trials, should be feasibly implemented to overcome the presented challenges.

Fluid Shear Stress Promotes Autophagy in Hepatocellular Carcinoma Cells.

The autophagy in cancer cells is recognized as an essential hallmark of tumors, which can enhance cancer cell migration and invasion, and result in high incidence of tumor metastasis. The fluid shear stress (FSS) in tumor mechanical microenvironment plays a pivotal role in mediating the behaviors and functions of cells. In this study, the hepatocellular carcinoma cells were exposed to 1.4 dyn/cm2 FSS to explore whether FSS could induce autophagy. The results of TEM, Ad-mCherry-GFP labeled LC3B, and mRNA and protein expression of autophagy markers confirmed that FSS could induce autophagy in a time-dependent manner. Additionally, the inhibition of autophagy significantly downregulated the expression of PI3K, FAK and Rho GTPases, and attenuated the ability of cell migration, suggesting that FSS-induced autophagy depended on PI3K- FAK-Rho GTPases pathway. This study elucidated the role of FSS in inducing autophagy during tumor progression, which has emerged as a promising clinical strategy for cancer.