Smurf1-targeting microRNA-136-5p-modified bone marrow mesenchymal stem cells combined with 3D-printed ß-tricalcium phosphate scaffolds strengthen osteogenic activity and alleviate bone defects.

Suitable biomaterials with seed cells have promising potential to repair bone defects. However, bone marrow mesenchymal stem cells (BMSCs), one of the most common seed cells used in tissue engineering, cannot differentiate efficiently and accurately into functional osteoblasts. In view of this, a new tissue engineering technique combined with BMSCs and scaffolds is a major task for bone defect repair. Lentiviruses interfering with miR-136-5p or Smurf1 expression were transfected into BMSCs. The effects of miR-136-5p or Smurf1 on the osteogenic differentiation (OD) of BMSCs were evaluated by measuring alkaline phosphatase activity and calcium deposition. Then, the targeting relationship between miR-136-5p and Smurf1 was verified by bioinformatics website analysis and dual luciferase reporter assay. Then, a rabbit femoral condyle bone defect model was established. miR-136-5p/BMSCs/ß-TCP scaffold was implanted into the defect, and the repair of the bone defect was detected by Micro-CT and HE staining. Elevating miR-136-5p-3p or suppressing Smurf1 could stimulate OD of BMSCs. miR-136-5p negatively regulated Smurf1 expression. Overexpressing Smurf1 reduced the promoting effect of miR-136-5p on the OD of BMSCs. miR-136-5p/BMSCs/ß-TCP could strengthen bone density in the defected area and accelerate bone repair. SmurF1-targeting miR-136-5p-modified BMSCs combined with 3D-printed ß-TCP scaffolds can strengthen osteogenic activity and alleviate bone defects.

Molecular Link in Flavonoid and Amino Acid Biosynthesis Contributes to the Flavor of Changqing Tea in Different Seasons.

The present study was aimed to elucidate the flavor formation mechanism of Changqing tea. High-performance liquid chromatography (HPLC) analysis showed that the total catechins of Changqing tea was 65-160 mg/g, with 16-34 mg/g non-galloyated catechins and 49-126 mg/g galloylated catechins. Tea polyphenols and free amino acids account for 286-312 mg/g and 35-89 mg/g, respectively. Transcriptome of Changqing tea during different seasons revealed 316, 130 and 12 DEGs in comparisons of spring vs. autumn, spring vs. summer, and summer vs. autumn, respectively. Compared to spring, the genes involved in flavonoid biosynthesis and bitter imparted amino acids were up-regulated in summer and autumn. Metabolome analysis was conducted by using HPLC-MS; the result indicated that umami and kokumi contributing amino acids were decreased in summer and autumn compared with spring. It could be concluded that the coordination of flavonoid biosynthesis and amino acids biosynthesis resulted in the special flavor of Changqing tea.

Abnormal glucose regulation in Chinese patients with coronary artery disease: a gender analysis.

BACKGROUND: Diabetes and impaired glucose regulation are very common in patients with coronary artery disease (CAD). In this study, we aim to investigate the prevalence of abnormal glucose regulation in men and women in Chinese CAD patients. METHODS: In this retrospective study, 4100 patients (male, n = 2873; female, n = 1227)with CAD were enrolled. The mean age of these patients was 63 years. The demographic data, medical history, echocardiography findings and blood investigations were collected and analyzed. RESULTS: In this population, 953 (24%) patients had definite diagnosis of type 2 diabetes mellitus, including 636 males (23%) and 317 females (27%). There was a higher prevalence of diabetes in females than men (p < 0.05). For the remaining patients, 48% (n = 959) undergone an oral glucose tolerance test (OGTT), which revealed that 83 male patients (12%) and 41 female patients (16%) suffered from the type 2 diabetes (p > 0.05). 283 men (40%) and 105 women (41%) had impaired glucose regulation (IGR) (p > 0.05). Only 338 men (25%) and 109 women (19%) showed the normal glucose regulation, implying a higher prevalence of abnormal glucose regulation in females (p < 0.01). The odd ratio (OR) showed that women were more prone to have diabetes mellitus or IGT than men and the OR was 1.44 and 1.43 respectively. CONCLUSION: Abnormal glucose regulation is highly prevalent in CAD patients. The women are more prone to have diabetes mellitus or IGT than men.

Circular RNA_ANKIB1 accelerates chemo-resistance of osteosarcoma via binding microRNA-26b-5p and modulating enhancer of zeste homolog 2.

Osteosarcoma is a common bone malignancy in children and adolescents. Chemotherapeutic drug resistance is the major factor impacting the surgical outcome and prognosis of patients with osteosarcoma. This investigation assessed the role and mechanism of circular RNA_ANKIB1 in the development of osteosarcoma. The circular RNA (circ) _ANKIB1, microRNA (miR)-26b-5p, enhancer of zeste homolog 2 (EZH2) expression in OS samples was investigated through RT-qPCR. The EZH2, multidrug resistance protein 1 (MRP1), P-gp, and lipoprotein receptor-related protein (LRP) protein expressions were analyzed through western blot. The association between circ_ANKIB1 and the occurrence of clinic-pathological features in OS patients was assessed; the circular features of circ_ANKIB1 were analyzed. The hFOB1.19, KHOS, U2-OS OS cells were used to study the semi-inhibitory concentration IC50 of Doxorubicin (DXR)-resistant cells, clone formation, invasion, and apoptosis. The luciferase assay was used to study the binding of circ-ANKIB1 with miR-26b-5p and the targeting of miR-26b-5p with EZH2. In vivo experiments were performed via subcutaneous tumorigenic experiments. MiR-26b-5p in OS tissues and cells and DXR-resistant OS tissues and cells was silenced while circ_ANKIB1 and EZH2 were elevated. Circ_ANKIB1 silencing elevated miR-26b-5p, repressed EZH2, MRP1, P-gp, LRP, IC50, and elevated OS advancement. Circ_ANKIB1 bind miR-26b-5p. Reduced miR-26b-5p revered the influence of silencing circ_ANKIB1 on DXR resistant OS cells. MiR-26b-5p targeted EZH2, and EZH2 elevation reversed the impact of increasing miR-26b-5p on DXR resistant cells. Circ_ANKIB1 silencing suppressed DXR-resistant OS cells in vivo. In conclusion, Circ_ANKIB1 binds miR-26b-5p and modulates EZH2 to accelerate the chemo-resistance of osteosarcoma.

Fat mass and obesity associated (FTO)-mediated N6-methyladenosine modification of Krüppel-like factor 3 (KLF3) promotes osteosarcoma progression.

ARSTRACTN6-methyladenosine (m6A) methylation is the most common and abundant methylation modification of eukaryotic mRNAs, which is involved in tumor initiation and progression. The study aims to explore the potential role and the regulatory mechanism of fat mass and obesity associated (FTO) in osteosarcoma (OS) progression. In this study, we detected the expressions of Krüppel-like factor 3 (KLF3) in OS cells and tissues and found that the mRNA and protein levels of KLF3 were increased in OS cells and tissues and significantly related to tumor size, metastasis, and TNM stage and poor prognosis of OS patients. FTO promoted the proliferation and invasion and suppressed apoptosis of OS cells through cell experiments in vitro. Further mechanism dissection revealed that FTO and YTHDF2 enforced the decay of KLF3 mRNA and decreased its expression. FTO-mediated mRNA demethylation inhibited KLF3 expression in the YTHDF2-dependent manner. Moreover, KLF3 overexpression abrogated FTO-induced oncogenic effects on the proliferation and invasion of OS cells. Overall, our findings showed that FTO-mediated m6A modification of KLF3 promoted OS progression, which may provide a therapeutic target for OS.

Long non-coding RNA long intergenic non-coding 00641 mediates cell progression with stimulating cisplatin-resistance in osteosarcoma cells via microRNA-320d/myeloid cell leukemia-1 axis.

As a staple chemotherapy medicine, cisplatin (DDP) is extensively applied in cancer patients, but its drug resistance is limited. Numerous studies have elucidated that long non-coding RNA (lncRNA) performs as a pivotal agent in osteosarcoma (OS). Nevertheless, lncRNA long intergenic non-coding 00641 (LINC00641)'s functions in DDP resistance for OS remain obscure. The purpose of this study was to investigate the effect and mechanism of LINC00641 on drug resistance of OS. The tissues of both clinical cancer patients and the normal control were gathered. Detection of LINC00641, microRNA-320d (miR-320d) and myeloid cell leukemia-1 (MCL1) was conducted. After the selection of OS cell lines, the detection of cell advancement was applied. Series of experiments were conducted to verify the interaction of LINC00641, miR-320d and MCL1. Xenografted tumor model in vivo was utilized to determine the function of LINC00641. The data displayed, LINC00641 was prominently elevated in OS tissues and cells, especially in DDP-resistant tumors and cell lines. Knock-down LINC00641 was able to attenuate progression of DDP-resistant OS cells thus dampening their drug resistance toward DDP. Moreover, knock-downing LINC00641 gene was also able to manifest antagonism toward DDP-resistance in vivo. On the grounds of bioinformatics prediction, a direct binding of LINC00641 with miR-320d existed, whose target was MCL1. Meanwhile, LINC00641 modulated MCL1 via targeting miR-320d. Additionally, repressive LINC00641 blocked MCL1 via emulative interaction with miR-320d, thus expediting DDP-sensitivity of OS cells. All in all, it is found that LINC00641 is available to escalate drug resistance of DDP-resistant OS cells via mediation of miR-320d/MCL1 axis.

Downregulation of MIAT reduces the proliferation and migratory and invasive abilities of retinoblastoma cells by sponging miR-665 and regulating LASP1.

Long non-coding RNAs (lncRNAs) can function as onco-lncRNAs in several types of human cancer, including retinoblastoma (Rb). The present study investigated the potential role and regulatory mechanism of the lncRNA myocardial infarction-associated transcript (MIAT) in Rb. To do so, the expression levels of MIAT, microRNA (miR)-665, and LIM and SH3 protein 1 (LASP1) in Rb tissues from patients or Rb cells were analysed using reverse transcription quantitative PCR. The interactions between miR-665 and MIAT/LASP1 were confirmed by the dual-luciferase reporter assay. MTT, Transwell (to assess migration and invasion) and western blotting assays were used to explore the functions of the MIAT/miR-665/LASP1 axis on Rb progression in vitro. The results of the present study indicated that MIAT targeted miR-665. In Rb tissues and cell lines, high expression of MIAT was observed, whereas miR-665 was downregulated in Rb tissues. Furthermore, the proliferation and migratory and invasive abilities of Rb Y79 and HXO-RB44 cells were decreased following MIAT downregulation or miR-665 overexpression. In addition, LASP1 was identified as a target gene of miR-665. Both the decreased expression of miR-665 and the elevated expression of LASP1 reversed the suppressive effects of MIAT knockdown on the proliferation and migratory and invasive abilities of Y79 cells. Furthermore, MIAT silencing attenuated the development of Rb by regulating the miR-665/LASP1 axis. Taken together, these findings suggested that MIAT may be considered as a possible therapeutic target for Rb.

miR-27a promotes osteogenic differentiation in glucocorticoid-treated human bone marrow mesenchymal stem cells by targeting PI3K.

MicroRNA-27a (miR-27a) modulates osteogenic differentiation (OD); however, the mechanism by which it influences osteoclastic activity in the glucocorticoid (GC)-elicited osteoporotic bone is still unclear. Bone marrow was obtained from the proximal femur of patients (n = 3) with a femoral neck fracture and those (n = 3) with steroid-related osteonecrosis of the femoral head (ONFH). GC was applied to an established ONFH cell model from human bone marrow mesenchymal stem cells (hBMSCs). The miR-27a expression profiles were found to be downregulated in ONFH samples and GC-induced hBMSCs using microarray analysis and real-time quantitative polymerase chain reaction, whereas the OD capacity of hBMSCs was significantly reduced in the GC group compared with the control group. Subsequent transfection of an miR-27a mimic in hBMSCs revealed that the OD capacity of cells was remarkably strengthened in the GC group compared with the miR-control group. Bioinformatics software (TargetScan) predicted that phosphoinositide 3-kinase (PI3K) might be a potential miR-27a target, which was indicated by dual-luciferase reporter assay. Compared with the control group, the GC group exhibited a significantly downregulated protein expression level of PI3K and its downstream protein kinase B (Akt) and mammalian target of rapamycin (mTOR) expression. Furthermore, administration of 10 µM 740 Y-P, a cell-permeable phosphopeptide activator of PI3K, to hBMSCs increased the expression of Akt and mTOR. Treatment with 740 Y-P reversed the effect of miR-27a on OD in hBMSCs. In conclusion, miR-27a is thought to relieve ONFH and the OD repression in GC-induced hBMSCs by targeting the PI3K/Akt/mTOR pathway.

microRNA-199a counteracts glucocorticoid inhibition of bone marrow mesenchymal stem cell osteogenic differentiation through regulation of Klotho expression in vitro.

Osteogenic differentiation (OD) of bone marrow mesenchymal stem cells (BMSCs) is critically important for mitigation of osteoporosis. Glucocorticoids (GCs) are extensively used for treating chronic inflammation, although long-term exposure to GCs is capable of triggering osteoporosis. microRNAs (miRNAs) have been reported to play a critical role in bone diseases. In the present study, we treated BMSCs with dexamethasone (DEX) during OD to stimulate GC-mediated osteoporosis. Microarray and quantitative polymerase chain reaction (Q-PCR) assays demonstrated that miR-199a was upregulated during OD of BMSCs, while DEX treatment caused a significant reduction in miR-199a. Alkaline phosphatase (ALP) activity, Alizarin red (AR) staining, and Q-PCR were applied to assess the role of miRNA-199a overexpression in DEX-triggered OD inhibition. miR-199a was able to rescue OD and ALP activity, which were inhibited by DEX. Additionally, we observed that ALP, BMP2, COL1A1, and Runx2 were increased after transfection of miRNA-199a mimics. Furthermore, we confirmed that miRNA-199a facilitates OD of BMSCs through direct inhibition of Klotho protein and messenger RNA expression affecting the downstream fibroblast growth factor receptor 1/extracellular-signal-regulated kinase and Janus kinase 1/signal transducer and activator of transcription 1 pathways. This study indicates that miR-199a plays a critical role in preventing GC-mediated osteoblast differentiation and may function as a promising miRNA biomarker for osteoporosis.

MiR-138 Acts as a Tumor Suppressor by Targeting EZH2 and Enhances Cisplatin-Induced Apoptosis in Osteosarcoma Cells.

Chemotherapeutic insensitivity remains a major obstacle to treating osteosarcoma effectively. Recently, increasing evidence has suggested that microRNAs (miRNAs) are involved in drug resistance. However, the effect of miR-138 on cisplatin chemoresistance in osteosarcoma has not been reported. We used real-time PCR to detect the expression of mature miR-138 in osteosarcoma tissues and cell lines. Cell proliferation, invasion, and migration assays were used to observe changes to the osteosarcoma malignant phenotype. MiR-138 was downregulated in osteosarcoma tissues and cell lines, and miR-138 overexpression negatively regulated osteosarcoma cell proliferation, migration, and invasion. We also verified that EZH2 is a direct target of miR-138. Furthermore, enhancing EZH2 expression reduced the inhibitory effects of miR-138 on osteosarcoma. Proliferation, apoptosis assays and caspase-3 activity assay confirmed that elevated miR-138 expression enhanced osteosarcoma cell chemosensitivity to cisplatin by targeting EZH2. In conclusion, the present study demonstrates that miR-138 acts as a tumor suppressor by enhancing osteosarcoma cell chemosensitivity and supports its potential application for treating osteosarcoma in the future.

Anti-inflammatory flavonolignans from Hydnocarpus anthelminthica seeds.

A new flavonolignan, anthelminthicol A (1), together with four known compounds, was isolated from the EtOAc extracts of the seeds of Hydnocarpus anthelminthica. Their structures were elucidated using extensive spectroscopic techniques. Bioassay showed that compounds 3-5 could inhibit nitric oxide production in LPS-stimulated RAW 264.7 macrophage cell lines, with IC(50) values of 7.81, 9.38, and 10.55 µM, respectively.

[Mammography and magnetic resonance imaging for diagnosis of the intraductal papilloma of the breast].

OBJECTIVE: To investigate the features of intraductal papilloma of the breast in mammography and magnetic resonance imaging (MRI) and assess the diagnostic values of the two imaging modalities. METHODS: Fifteen patients with intraductal papilloma of the breast confirmed surgically and pathologically underwent X-ray examination of the breast, and 11 of them also received enhanced MRI. The imaging findings by mammography and MRI were compared. RESULTS: Enhanced MRI clearly displayed the location and morphology of the intraductal papilloma, and 7 patients showed smooth tumor margins and 2 showed irregular margins. On T(1)WI, the lesions were isointense or slightly hypointense, and appeared isointense or slightly hyperintense on T(2)WI. Some of the intraductal papillomas were seen encapsulated in the dilated ductal. The varying enhancement features of the lesions increased the difficulty in distinguishing from carcinoma. Mammography identified intraductal papillomas only in 2 of the 15 cases (13%) with lesion feature similar to that found by MRI. Fine cluster calcification was found in 1 case. CONCLUSION: MRI can more accurately define the location of the lesion than X-ray. In spite of some resemblance in the MRI findings between intraductal papillomas and breast carcinoma, MRI still serves as a useful diagnostic modality for intraductal papilloma that shows some characteristic findings.

Esthesioneuroblastoma methods of intracranial extension: CT and MR imaging findings.

INTRODUCTION: Esthesioneuroblastoma (ENB) is an aggressive neuroectodermal malignancy in the upper nasal cavity with local infiltration and lymphatic or hematogenous metastasis. The purpose of this paper is to document three types of direct intracranial extensions by ENB using computed tomography (CT) and magnetic resonance imaging (MRI). METHODS: Eleven patients with pathologically confirmed ENB were admitted in our hospital between December 2002 and December 2008. Their magnetic resonance (MR; n = 10) and CT (n = 8) images were retrospectively reviewed, and particular attention was paid to tumor location and extension, enhancement pattern, cervical lymph node metastasis, and Kadish stage. RESULTS: The majority of patients were male (8/11) with Kadish stage C tumor (10/11). Three types of direct intracranial extension by ENBs were put forward according to their MR and CT findings. The primary tumors were well-defined soft-tissue masses centered in the roof of the nasal cavity eroding into the paranasal sinuses (11/11), the contralateral nasal cavity (4/11), the cranial cavity (5/11), and the fossa orbitalis (3/11). The tumor parenchyma were hypointensity on T1-weighted images, heterogeneous hyperintensity on T2-weighted images, and isodensity or slight hyperdensity on CT images with scattered necroses (4/11) and marginal cysts(4/11). Their enhancements were significant and inhomogeneous. Cervical lymph nodes metastases were observed in four patients (4/11), but no pathologically proved distant metastasis was observed. CONCLUSION: Three types of direct intracranial extensions by ENB can be found on CT and MRI: cranio-orbital-nasal-communicating ENB, cranio-nasal-communicating ENB, and orbital-nasal-communicating ENB.

[Establishment of a rabbit model bearing transplanted endometrial carcinoma and magnetic resonance imaging features of the metastatic lymph nodes].

OBJECTIVE: To establish a rabbit model bearing endometrial carcinoma and observe the magnetic resonance imaging (MRI) features of the metastatic lymph nodes. METHODS: VX(2) tumor grafts were orthotopically embedded into the endometrium of the rabbits. Three weeks after the implantation, the tumor and the metastatic retroperitoneal lymph nodes were examined with MRI, and the signal intensities and the size of the lymph nodes were compared with those in normal rabbits. RESULTS: Orthotopic tumor growth was observed in all the rabbits. Tumor infiltration of the serosa and retroperitoneal lymph node metastasis occurred 3 weeks after tumor implantation. MRI demonstrated obviously lymph node enlargement in the tumor-bearing rabbits as compared with those of normal rabbits, while the signal intensity of the lymph nodes were comparable between them. CONCLUSION: The endometrial carcinoma in this rabbit model well simulate the metastatic behavior of human endometrial carcinoma, and may serve as a good model for testing the efficacy of the contrast agents for MRI of the lymph nodes.

[CT diagnosis and pathological findings of testicular lesion].

OBJECTIVE: To investigate the value of CT scan in the diagnosis and differential diagnosis of testicular space-occupying lesions. METHODS: In patients with testicular space-occupying lesions, 11 underwent examinations for such tumor markers as alpha-fetoprotein (AFP) and human chorionic gonadotropin (HCG). Plain CT scans were performed with a section thickness and interval of 2 mm followed by enhanced scans after intravenous bolus injection of 70 ml 60% ultravist. Spiral CT multiplanar reconstruction images were also obtained. RESULTS: Of the 13 patients, histopathologically confirmed seminoma was found in 3, endodermal sinus tumor in 1, teratoma in 2, mixed germinoma in 2, leucoma in 1 case, abscess in 2, and tuberculosis in 2. One seminoma patient had slightly elevated AFP level while 5 of the 6 noneminoma patients (83%) had elevated AFP level. CT scans displayed characteristic features for diagnosis of testicular space-occupying lesions. Spiral CT with multiplanar reconstruction allowed full view of the lesions and demonstrated their relationship with the surrounding structures. CONCLUSION: CT scan in combination with the clinical analysis and tumor marker examinations can be of important value in the diagnosis and differential diagnosis of testicular space-occupying lesions. The application of spiral CT multiplanar reconstruction also helps in the diagnosis.

[Comparison of digital radiography and conventional X-ray in the diagnosis of solitary pulmonary nodule with receiver operating characteristic analysis].

OBJECTIVE: To compare digital radiographs with conventional radiographs in the detection of solitary pulmonary nodules. METHODS: Thirty patients with solitary pulmonary nodule and 30 cases without pulmonary nodules were enrolled in the study. The existence of solitary pulmonary nodule was confirmed by chest computed tomography (CT) as well as biopsy. All patients examined by both digital radiography (group A) and conventional radiographs (group B) were reviewed by four experienced chest radiologists and four residents. Assessment was performed with receiver operating characteristic (ROC) analysis of the images in both groups. RESULTS: (1) Observer performance of the experienced radiologists in group B (Az=0.838) was superior to that in group A (Az=0.816) (P<0.05) in detection of solitary pulmonary nodule. For the residents, observer performance in group B (Az=0.842) was superior to that in group A (Az=0.712) (P<0.05). (2) There was no difference between the two groups (P=0.272), for the judgement of benign or malignant solitary pulmonary nodule. CONCLUSIONS: Digital radiographs is superior to conventional radiographs in detection of solitary pulmonary nodule. However, there was no significant differences in discrimination between benign and malignant solitary pulmonary nodules in the two groups.

[Mammographic findings of gynecomastia].

OBJECTIVE: To investigate the features of gynecomastia displayed by mammography. METHODS: Twelve patients with gynecomastia were examined with a high-performance GITTO-TECH mammograph (IMS Company, Italy), and the results were compared with those obtained from pathological examination. RESULTS: The 12 cases were pathologically confirmed as gynecomastia, 10 of which were also identified by mammography while 2 misdiagnosed as male breast cancer. CONCLUSION: Diagnosis of gynecomastia can be established when typical features are presented in mammography, and fine needle aspiration biopsy can be performed when possible for discrimination from male breast cancer.