Depletion of circRNA circ_CDK14 inhibits osteosarcoma progression by regulating the miR-520a-3p/GAB1 axis.

Osteosarcoma (OS) is a lethal bone malignancy. Circular RNAs (circRNAs) have emerged as important regulators of OS development. CircRNA cyclin dependent kinase 14 (circ_CDK14) was reported to be a potential oncogene in OS. However, the mechanistic pathway by which circ_CDK14 functions in OS is largely unknown. The relative expression of circ_CDK14, microRNA (miR)-520a-3p, and GRB2 Associated Binding Protein 1 (GAB1) was evaluated by quantitative real-time PCR and western blot assays. Flow cytometry was employed to monitor cell cycle distribution and apoptosis. Methyl thiazolyl tetrazolium (MTT) and colony formation assays were performed to assess cell viability and colony formation ability, respectively. Western blot assay was also used to detect the expression of apoptosis-related proteins. Transwell assay was carried out to monitor cell migration and invasion. Additionally, the target association between miR-520a-3p and circ_CDK14 or GAB1 was confirmed by a dual-luciferase reporter assay. Xenograft assay was applied to investigate the role of circ_CDK14 in vivo. Circ_CDK14 and GAB1 expression was upregulated, while miR-520a-3p was downregulated in OS tissues and cells. Circ_CDK14 depletion hindered OS cell proliferation, metastasis, and tumorigenesis while facilitated apoptosis, which were all ameliorated by miR-520a-3p inhibition. Circ_CDK14 could sponge miR-520a-3p. miR-520a-3p targeted GAB1 to repress OS cell proliferation and metastasis. Circ_CDK14 knockdown blocked OS tumor growth in vivo. Circ_CDK14 might positively affect OS development by modulating the miR-520a-3p/GAB1 axis.

Roles of miR-182 in Intervertebral Disc Degeneration and its Potential Prognostic Value.

BACKGROUND: MicroRNAs were reported to be involved in the progression of intervertebral disc degeneration (IDD). This study focused on the potential prognostic value and the underlying mechanism of miR-182 in IDD. METHODS: The expression level of miR-182 in plasma samples from 60 IDD patients and 60 healthy controls were examined in the present study. Then, the relationship between miR-182 expression and clinical features of IDD patients was analyzed. Moreover, the nucleus pulposus (NP) cells were cultured and transfected with either miR-182 inhibitor, mimics, or NC to explore the effects of miR-182 on cell proliferation and apoptosis. Furthermore, expression levels of proliferation and apoptosis-related proteins Bcl-2, Bax, and Caspase-3 were also evaluated. RESULTS: The expression level of miR-182 was dramatically increased in plasma samples of IDD patients compared with the controls. Moreover, ROC analysis indicated that miR-182 was a feasible diagnostic indicator for the diagnosis of IDD. According to the Japanese Orthopaedic Association (JOA) score, the prognosis of patients with the lower expression levels of miR-182 was better than for those with the higher expression levels of miR-182 in IDD. Furthermore, the miR-182 inhibitor significantly increased the proliferation, decreased the apoptosis of human NP cells, and altered the expression of proliferation and apoptosis-related proteins Bcl-2, Bax, and Caspase-3. On the contrary, miR-182 mimics notably inhibited proliferation but promoted apoptosis of human NP cells and increased Bax and Caspase-3 expressions while reducing the Bcl-2 level. CONCLUSIONS: miR-182 was negatively correlated with the prognosis of the IDD patients and affected the proliferation and apoptosis of NP cells in vitro.

TiRobot-Assisted Percutaneous Cannulated Screw Fixation in the Treatment of Femoral Neck Fractures: A Minimum 2-Year Follow-up of 50 Patients.

OBJECTIVE: To assess the long-term clinical efficacy of TiRobot-assisted percutaneous cannulated screw fixation in the treatment of femoral neck fractures. METHODS: This retrospective study included 50 patients with unilateral femoral neck fractures who were treated with TiRobot-assisted percutaneous cannulated screw fixation from September 2017 to May 2018. After at least 2 years of follow-up, the results of treatment, including operation duration, frequency of fluoroscopy use, intraoperative bleeding, hospital stay, medical expense, screw placement accuracy, rate of fracture healing and necrosis of the femoral head, and Harris hip scores at the last follow up, were recorded and compared with those of 83 matched patients who underwent conventional manual positioning surgery. RESULTS: The TiRobot group had longer operation duration (83.3 ± 31.2 min vs 44.1 ± 14.8 min) and higher medical expenses (28,407.1 ± 7498.0 yuan vs 22,672.3 ± 4130.3 yuan) than the conventional group. The TiRobot group had significantly less intraoperative bleeding (11.3 ± 7.3 mL vs 51.6 ± 40.4 mL) and shorter hospital stay (8.6 ± 2.8 days vs 11.1 ± 3.41 days) than the conventional group. Screw parallelism (1.32° ± 1.85° vs 2.54° ± 2.99° on anteroposterior radiograph; 1.42° ± 2.25° vs 3.09° ± 3.63° on lateral radiograph) and distance between screws (58.44 ± 10.52 mm vs 39.69 ± 12.17 mm) were significantly improved. No significant difference was found between the two groups in terms of the use of fluoroscopy (40.1 ± 28.5 times vs 38.6 ± 21.0 times) and Harris hip scores at the last follow-up (93.2 ± 10.3 points vs 88.4 ± 11.9 points). Two cannulated screws penetrated the femoral head during manual insertion in the conventional group but not in the TiRobot group. The rate of nonunion and necrosis of the femoral head in the TiRobot group was reduced compared with that in the conventional group (0 vs 7.2%; 6.0% vs 24.1%). CONCLUSION: TiRobot-assisted percutaneous cannulated screw fixation of femoral neck fractures is accurate and minimally invasive and helps in reducing late complications, particularly necrosis of the femoral head and nonunion of fractures.

Upregulation of suppressor of cytokine signaling 3 ameliorates spinal degenerative disease in adolescents by mediating leptin and tumor necrosis factor-α levels.

Spinal degenerative changes may occur following the rapid growth observed in adolescents, causing a reduced quality of life. The suppressor of cytokine signaling (SOCS) is involved in various degenerative diseases. The current study recruited adolescents with spinal degenerative disease (SDD) to identify the effect of SOCS-3 on leptin and tumor necrosis factor-α (TNF-α) levels in this disorder. From January 2010 to January 2016, 120 adolescents (aged 14 to 25) were enrolled in the current study, with 68 diagnosed with SDD and the remaining 52 treated as controls. Nucleus pulposus cells (NPCs) were extracted and cultured in vitro. TNF-α levels in NPCs were determined using flow cytometry. Degenerative NPCs were then transfected with pCR3.1-SOCS-3 and ELISA was performed to determined TNF-α and leptin levels. RT-qPCR was performed to measure the mRNA level of SOCS-3 and leptin in NPCs and western blotting was utilized to detect the protein level of leptin and the extent of leptin receptor phosphorylation. The results revealed that TNF-α levels in degenerative NPCs were higher than those in normal NPCs. The overexpression of SOCS-3 reduced levels of TNF-α and leptin in degenerative NPCs. In addition, the upregulation of leptin increased SOCS-3 levels in a concentration-dependent manner. Furthermore, the expression of the leptin receptor and phosphorylated leptin receptor gradually decreased with increasing leptin concentrations and the level of phosphorylated leptin receptor negatively correlated with SOCS-3 expression. The inductive effect of leptin on the level of SOCS-3 and the inhibitory effect of SOCS-3 on the activity of leptin were identified. The current study demonstrated that SOCS-3 reduces leptin and TNF-α levels in degenerative NPCs from adolescents, indicating its potential role in the development of novel SDD therapies.

Thermal pretreatment promotes the protective effect of HSP70 against tendon adhesion in tendon healing by increasing HSP70 expression.

Tendon adhesion is a substantial challenge for tendon repair. Thermal pretreatment (TP) may decrease inflammation by upregulating heat shock proteins (HSPs). The present study intends to identify the function that TP serves when combined with HSP70 overexpression in tendon healing and adhesion in rats. Sprague­Dawley male rats were used to establish a surgically ablative tendon postoperative suture model, and the positive expression of the HSP70 protein was measured using immunohistochemistry. Changes to the blood vessels and collagenous fiber, in addition to the maximum tensile strength and the tendon sliding distance, were detected under a microscope. Finally, HSP70, tumor growth factor ß (TGF­ß), and insulin­like growth factor 1 (IGF­1) mRNA and protein levels were all determined by employing reverse transcription­quantitative polymerase chain reaction and western blot analysis methods. The positive expression of the HSP70 protein increased following TP. Furthermore, TP reduced the infiltration of inflammatory cells and improved the collagenous arrangement, accompanied by an increased maximum tensile force and tendon gliding distance following surgery. In addition, TP increased the mRNA and protein expression levels of HSP70, TGF­ß and IGF­1. Altogether, TP increases HSP70 expression, thereby reducing postoperative traumatic inflammation and establishing tendon adhesion and promoting tendon healing. Thus, TP may be a potential strategy for the treatment of tendon adhesion.

Down-regulation of long non-coding RNA ITGB2-AS1 inhibits osteosarcoma proliferation and metastasis by repressing Wnt/ß-catenin signalling and predicts favourable prognosis.

Osteosarcoma is the most common and highly aggressive bone neoplasm often occurs among adolescents and young people. In despite of the major advances in the treatment, the overall survival of osteosarcoma patients remains poor. Long non-coding RNAs (lncRNAs) have been identified as key regulators involved in tumorigenesis and progression of osteosarcoma. However, the exact roles and mechanisms of lncRNAs in osteosarcoma remain unclear. In this study, the functions and underlying molecular mechanisms of a novel lncRNA, ITGB2 antisense RNA1 (ITGB2-AS1) were investigated in osteosarcoma. The expression levels of ITGB2-AS1 were up-regulated in osteosarcoma tissue samples and cell lines determined by qRT-PCR assay. Osteosarcoma patients with high ITGB2-AS1 expression had a significantly poor prognosis. In addition, knockdown of ITGB2-AS1 inhibited the proliferation and induced apoptosis of osteosarcoma cells using MTT assay and flow cytometry detection. Furthermore, wound healing and transwell invasion assay revealed that depletion of ITGB2-AS1 suppressed the migration and invasion abilities of osteosarcoma cells. Molecular mechanism study indicated that ITGB2-AS1 inhibition impaired Wnt/ß-catenin signalling pathway in osteosarcoma cells. Taken together, our study suggested that ITGB2-AS1 played critical roles in the development and progression of osteosarcoma via Wnt/ß-catenin signalling, indicating that ITGB2-AS1 might be a valuable diagnostic biomarker and potential anticancer therapeutic target for osteosarcoma.

The Mitochondrial Na+/Ca2+ Exchanger is Necessary but Not Sufficient for Ca2+ Homeostasis and Viability.

Luongo et al. found that the mitochondrial Na+/Ca2+ exchanger (NCLX) was essential for Ca2+ homeostasis and viability. Here, we re-analyze their data in terms of fractal self-similarity and quantitative difference (QD). We calculated the 7-dimension data from NCLX conditional loss-of-function mouse models, and the 9-dimension data from NCLX overexpression (NCLX-Tg) models. RESULTS: The 9-dimension data of the NCLX-Tg and its tTA control were partially self-similar to each other, while the 7-dimension data in NCLX knockout models were not. CONCLUSION: The NCLX may be necessary but is not sufficient for Ca2+ homeostasis and viability.

Mitochondria-targeted platinum(II) complexes induce apoptosis-dependent autophagic cell death mediated by ER-stress in A549 cancer cells.

Agents with multiple modes of tumor cell death can be effective chemotherapeutic drugs. One example of a bimodal chemotherapeutic approach is an agent that can induce both apoptosis and autophagic death. Thus far, no clinical anticancer drug has been shown to simultaneously induce both these pathways. Mono-functional platinum complexes are potent anticancer drug candidates which act through mechanisms distinct from cisplatin. Here, we describe the synthesis and characterize of two mono-functional platinum complexes containing 8-substituted quinoline derivatives as ligands, [PtL1Cl]Cl [L1 = (Z)-1-(pyridin-2-yl)-N-(quinolin-8-ylmethylene) methanamine] (Mon-Pt-1) and [PtL2Cl]Cl [L2 = (Z)-2-(pyridin-2-yl)-N-(quinolin-8-ylmethylene) ethanamine] (Mon-Pt-2). In comparison to cisplatin, Mon-Pt-2 exhibited a greater in vitro cytotoxicity, was more effective in resistant cells and elicited a better anticancer effect. Mechanistic experiments indicate that Mon-Pt-2 mainly accumulates in mitochondria, and stimulates significant TrxR inhibition ROS release and an ER stress response, mediated by mitochondrial dysfunction, ultimately resulting in a simultaneous induction of apoptosis and autophagy. Importantly, compared to cisplatin, Mon-Pt-2 exhibits lower acute toxicity and better anticancer activity in a murine tumor model. To the best of our knowledge, Mon-Pt-2 is the first mono-functional platinum complex inducing pro-death autophagy and apoptosis of cancer cells.

Organometallic Gold(III) Complexes Similar to Tetrahydroisoquinoline Induce ER-Stress-Mediated Apoptosis and Pro-Death Autophagy in A549 Cancer Cells.

Agents inducing both apoptosis and autophagic death can be effective chemotherapeutic drugs. In our present work, we synthesized two organometallic gold(III) complexes harboring C^N ligands that structurally resemble tetrahydroisoquinoline (THIQ): Cyc-Au-1 (AuL1Cl2, L1 = 3,4-dimethoxyphenethylamine) and Cyc-Au-2 (AuL2Cl2, L2 = methylenedioxyphenethylamine). In screening their in vitro activity, we found both gold complexes exhibited lower toxicity, lower resistance factors, and better anticancer activity than those of cisplatin. The organometallic gold(III) complexes accumulate in mitochondria and induce elevated ROS and an ER stress response through mitochondrial dysfunction. These effects ultimately result in simultaneous apoptosis and autophagy. Importantly, compared to cisplatin, Cyc-Au-2 exhibits lower toxicity and better anticancer activity in a murine tumor model. To the best of our knowledge, Cyc-Au-2 is the first organometallic Au(III) compound that induces apoptosis and autophagic death. On the basis of our results, we believe Cyc-Au-2 to be a promising anticancer agent or lead compound for further anticancer drug development.

Crystal structure, cytotoxicity and action mechanism of Zn(II)/Mn(II) complexes with isoquinoline ligands.

Four µ2-Cl bridged dinuclear metal complexes with isoquinoline ligands, (MPDQ)2Zn2Cl4 (1) (MPDQ=4.5-methylenedioxy-1-pyridinedihydroisoquinoline), (PYP)2Zn2Cl4 (2) (PYP=5-pyridin-2-yl-[1,3]dioxolo[4,5-g]isoquinoline), (MPDQ)2Mn2Cl4 (3),and (PYP)2Mn2Cl4 (4) were synthesized and characterized. All complexes exhibited strong proliferation inhibition activity against various cancer cells. The underlying molecular mechanisms through which they caused the cancer cell death were also elucidated. Induction of apoptosis in MGC-803 cells by complex 2 was evidenced by annexin V+/PI- detection and DiD/DAPI staining assay. Further investigation revealed that complex 2 was able to induce intrinsic pathway-dependent apoptosis in cancer cells by triggering DNA damage which was caused by the overproduction of reactive oxygen species. Based on these studies, we suggest that Zn(II) complexes containing isoquinoline ligands can be developed as candidates for anti-cancer chemotherapeutics.

[Effects of plastic film mulching and rain harvesting modes on chlorophyll fluorescence characteristics, yield and water use efficiency of dryland maize].

The differences on chlorophyll fluorescence parameters, yield and water use efficiency of dryland maize were compared among full plastic film mulching on double ridges and planting in catchment furrows (FFDRF), half plastic film mulching on double ridges and planting in catchment furrows (HFDRF), plastic film mulching on ridge and planting in film-side (FS), and flat planting with no plastic film mulching (NM) under field conditions in dry highland of Loess Plateau in 2007-2012. The results showed that fluorescence yield (Fo), the maximum fluorescence yield (Fm), light-adapted fluorescence yield when PS II reaction centers were totally open (F), light-adapted fluorescence yield when PS II reaction centers closed (Fm'), the maximal photochemical efficiency of PS II (Fv/Fm), the actual photochemical efficiency of PS II in the light (Phi PS II), the relative electron transport rate (ETR), photochemical quenching (qP) and non-photochemical quenching (qN) in maize leaves of FFDRF were higher than that of control (NM), and the value of 1-qP was lower than that of control, at 13:00, chlorophyll fluorescence parameters values of FFDRF was significantly higher than control, which were increased by 5.3%, 56.8%, 10.7%, 36.3%, 23.6%, 56.7%, 64.4%, 45.5%, 23.6% and -55.6%, respectively, compared with the control. Yield and water use efficiency of FFDRF were the highest in every year no matter dry year, normal year, humid year and hail disaster year. Average yield and water use efficiency of FFDRF were 12,650 kg x hm(-2) and 40.4 kg x mm(-1) x hm(-2) during 2007-2012, increased by 57.8% and 61.6% compared with the control, respectively, and also significantly higher compared with HFDRF and PS. Therefore, it was concluded that FFDRF had significantly increased the efficiency of light energy conversion and improved the production capacity of dryland maize.

[Echo-tracking technology for evaluating the impact of blood pressure on vascular endothelial function].

OBJECTIVE: To evaluate the effect of blood pressure on vascular endothelial function using echo-tracking (ET) technology. METHODS: Thirty hypertensive (HP) patients, 30 subjects with high normal blood pressure (HN), and 30 normotensive control (NC) subjects were enrolled in this study. For each subject, conventional two-dimensional ultrasound was performed to measure the intima-media thickness (IMT), and an ET system was utilized to assess the carotid elasticity (Ep, ß, AC, AI, and PWVß). RESULTS: As the blood pressure increased, IMT, Ep, ß, AI, and PWVß values all increased and AC value decreased. Before excluding the confounding factors, the difference in IMT, Ep, ß, AC, AI, and PWVß values were significant between the 3 groups. After excluding the confounding factors, only PWVß value was significantly different between HN group and NC group; but between HP and NC group and between HP and HN group, the other parameters still showed significant differences. Systolic blood pressure had significant influences on IMT, Ep, AC, AI, and PWVß values, diastolic blood pressure significantly affected AI value, and pulse pressure significantly affected Ep and ß values. CONCLUSION: High normal blood pressure has no obvious effects on vascular function, and blood pressure is an independent risk factor of vascular endothelial dysfunction only in the stage of early hypertention. In early atherosclerosis, systolic blood pressure is the most significant factors affecting vascular endothelial function, followed by pulse pressure and diastolic blood pressure.

Diagnosis and spectrum of melamine-related renal disease: plausible mechanism of stone formation in humans.

BACKGROUND: An epidemic of urinary stones affecting children after consumption of melamine tainted milk is unfolding. We defined clinicopathological features of the disease for diagnosis, monitoring, and treatment of this group of patients. METHODS: A clinicopathological study on exposed children with ultrasonographic evidence of urolithiasis was conducted. Melamine and cyanuric acid levels in the urine were determined by mass spectrometry. RESULTS: Disease severity varied from acute renal failure with hydronephrosis to symptomatic or asymptomatic stones with or without abnormal urinalysis. All cases were aged <3 y with >50% cases having predisposing urinary metabolic risk factors for urolithiasis. Most of the stones were located in the renal pelvis and measured 2.5-18 mm by ultrasonography. We found a strong correlation between renal stone size and urinary melamine concentration. For stones <10 mm, a 10 microg/mmol creatinine increase in urinary melamine concentration is associated with approximately 1 mm increase in the size of the stone. The high degree of correlation strongly suggests that melamine is related to stone formation in humans. Using ROC analysis, we propose that patients who have a persistent melamine level above the optimal cut-off value of 7.1 microg melamine/mmol creatinine in urine might have a significant exposure of melamine-tainted products. Unlike melamine, urinary cyanuric acid is not significantly different between cases and controls. Pathophysiological findings from feeding animals with melamine and cyanuric acid may not be directly applicable to humans. CONCLUSION: Both melamine and urine metabolic lithogenic factors are important for the formation of melamine-related stones. Apart from aiding with case screening and confirmation, the urine melamine level might as well be an indicator of residual melamine load in the body and thus is useful for following-up and monitoring of the confirmed cases. As the stones are small and can be passed out spontaneously, follow-up of these patients with urine melamine will be a convenient tool for monitoring the melamine load of the patients.

[Correlation between power Doppler vascularity index and microvessel density in high-grade gliomas and adjacent edema].

OBJECTIVE: To investigate the correlation between power Doppler vascularity index (PDVI) and microvessel density (MVD) and evaluate the angiogenesis in high-grade gliomas and the adjacent edema in patients with glioma using intraoperative power Doppler ultrasound (PDUS) during gross total resection. METHODS: In 25 cases of high-grade gliomas undergoing gross total tumor resections, PDUS was performed intraoperatively and the regions of interest within the tumor and the adjacent edema were analyzed with Photoshop software to measure the tumoral and peritumoral blood flow quantified as PDVI. The tumoral and adjacent MVD were determined using immunohistochemical staining for CD34. The correlation between PDVI in the gliomas and the adjacent edema and MVD in the corresponding areas were analyzed using Spearman correlation test. RESULTS: The measurement of both PVDI and MVD revealed significant difference in vascularity between the gliomas and the adjacent edema (t=0.000, P<0.01), and PDVI was positively correlated to MVD measurement (r=0.7248 in the tumors and r=0.6608 in the adjacent edema). CONCLUSIONS: The difference in the vascularity between the tumor and adjacent edema allows their distinction by PDUS during operation for high-grade glioma. Intraoperative PDUS provides an accurate and reliable means for measuring vascularity in the glioma and the adjacent edema tissue.

[Minimally invasive closure of atrial septal defect without extracorporeal circulation: report of 5 cases].

OBJECTIVE: To review our experience with a new surgical approach for closure of atrial septal defect (ASD) through a minimally invasive method without extracorporeal circulation. METHODS: Five female patients (age range, 7-38 years) with secondary atrial septal defect ranging from 1.8 to 3.4 cm, underwent minimally invasive closure under ultrasonic cardiography guidance. RESULTS: The ASD was closed successfully in all of the 5 cases without residual shunt or displacement of occluder. The mean hospital stay was 7 days. CONCLUSION: This minimally invasive surgical approach is an easy and safe voay for clinical application.

[Association of platelet endothelial cell adhesion molecule-1 gene polymorphism with coronary heart disease].

OBJECTIVE: To investigate the association of Leu125Val and Ser563Asn polymorphism of the gene encoding platelet endothelial cell adhesion molecule-1(PECAM-1) with coronary heart disease. METHODS: This study included 156 patients with the diagnoses of coronary heart disease (CHD) and coronary lesions derived from electrocardiography, myocardial enzyme analysis and coronary angiography as the CHD group, and another 75 in-patients admitted within the same period who showed no signs of CHD in the above examinations constituted the control group. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed to examine the missense polymorphism of PECAM-1gene in the position of Leu125Val and Ser563Asn. RESULTS: There were significant differences between CHD and control group in terms of the allele frequencies and genotype distributions of PECAM-1 gene, and the differences were especially conspicuous in the allele frequencies of 125Val and 563Asn (P<0.05) and genotype distributions of 125Val/Val and 563Asn/Asn. CONCLUSION: PECAM-1 gene polymorphism 1 may be a genetic risk factor for coronary heart disease.

[Association between aldosterone synthase gene polymorphism and hypertrophic cardiomyopathy].

OBJECTIVE: To investigate the relationship between aldosterone synthase (CYP11B2) gene polymorphism and hypertrophic cardiomyopathy (HCM). METHODS: Fifteen HCM patients and 18 healthy subjects were enrolled in this study. Peripheral blood samples were collected from these subjects to exact genome DNA. PCR and Hae III restriction endonuclease digestion were employed to study -344C/T polymorphism of CYP11B2 gene. RESULTS: CYP11B2 gene showed a significant difference in CT genotype distribution in HCM groups as compared with that in the control groups (P<0.05). CONCLUSION: CT genotype of CYP11B2 gene may be one of factors responsible for the pathogenesis of HCM in a proportion of patients.