The selective sponging of miRNAs by OIP5-AS1 regulates metabolic reprogramming of pyruvate in adenoma-carcinoma transition of human colorectal cancer.

RNA interactomes and their diversified functionalities have recently benefited from critical methodological advances leading to a paradigm shift from a conventional conception on the regulatory roles of RNA in pathogenesis. However, the dynamic RNA interactomes in adenoma-carcinoma sequence of human CRC remain unexplored. The coexistence of adenoma, cancer, and normal tissues in colorectal cancer (CRC) patients provides an appropriate model to address this issue. Here, we adopted an RNA in situ conformation sequencing technology for mapping RNA-RNA interactions in CRC patients. We observed large-scale paired RNA counts and identified some unique RNA complexes including multiple partners RNAs, single partner RNAs, non-overlapping single partner RNAs. We focused on the antisense RNA OIP5-AS1 and found that OIP5-AS1 could sponge different miRNA to regulate the production of metabolites including pyruvate, alanine and lactic acid. Our findings provide novel perspectives in CRC pathogenesis and suggest metabolic reprogramming of pyruvate for the early diagnosis and treatment of CRC.

Salvia Miltiorrhiza Bge.f.alba Ameliorates the Progression of Monocrotaline-Induced Pulmonary Hypertension by Protecting Endothelial Injury in Rats.

Pulmonary hypertension (PH) is a life-threatening disease that is characterized by elevated pulmonary blood pressure, abnormally thickened pulmonary arteries, and right ventricular hypertrophy. Monocrotaline (MCT) has been used to generate an experimental model of PH in rats, with PH initiated from injuries of lung vascular endothelium. Salvia Miltiorrhiza Bge.f.alba is a widely used traditional herb in China, known to exert protective effects on vascular endothelial cell injury in animal experiments. However, the role of Salvia Miltiorrhiza Bge.f.alba in PH remains unclear. Thus, we investigated the effects of the aqueous extract of Salvia Miltiorrhiza Bge.f.alba (AESM) on MCT-induced PH and explored the pertinent mechanism. PH was induced in rats by a single subcutaneous injection of MCT (60 mg/kg body weight). Low or high dose (4.6 g/kg or 14 g/kg body weight) of AESM was then administered orally for 21 days to PH rats. Hemodynamic study showed that AESM reduced mean pulmonary artery pressure and improved right ventricle function. Lung pathological analysis revealed that AESM reduced wall thickness and lumen stenosis of pulmonary vessels. Also AESM ameliorated right ventricular hypertrophy. Measurement of biochemical parameters indicated that AESM decreased endothelin-1 and thromboxane A2 in plasma and increased nitrogen monoxide and prostacyclin in the plasma and reduced the increase of transforming growth factor ß1 in lung tissue. Our results suggest that AESM may ameliorate the progression of MCT-induced PH in rats, at least in part by its protective effect on endothelial injury. Therefore, Salvia Miltiorrhiza Bge.f.alba could be useful in the treatment of PH.

Prognostic significance of BMP7 as an oncogene in hepatocellular carcinoma.

This study aims to evaluate the association between BMP7 tissue expression and patient prognosis in hepatocellular carcinoma (HCC). The expression of BMP7 mRNA in HCC was characterized using real-time PCR and 30 pairs of fresh frozen HCC tissues and corresponding noncancerous tissues. BMP7 protein expression in HCC was confirmed using immunohistochemistry on a tissue microarray chip. Finally, BMP7 expression was correlated with conventional clinicopathological features of HCC and patient outcome. The expression of BMP7 mRNA and protein in HCC cells was much higher than in normal hepatic cells. Our results showed that the high expression of BMP7 in HCC was related to tumor size (p < 0.001), histological differentiation (p = 0.041), serum AFP (p = 0.007), and tumor stage (p < 0.001). Kaplan-Meier survival analysis showed that a high-expression level of BMP7 resulted in a significantly poor prognosis of HCC patients. Multivariate analysis revealed that BMP7 expression level was an independent prognostic parameter for the overall survival rate of HCC patients. These findings provide evidence that a high-expression level of BMP7 serves as a biomarker for poor prognosis for HCC. Thus, we speculate that BMP7 may be a potential target of antiangiogenic therapy for HCC.

2,2'-Dichloro-N,N'-[1,3-phenyl-enebis(methyl-ene)]diacetamide.

The complete mol-ecule of the title compound, C(12)H(14)Cl(2)N(2)O(2), is generated by a crystallographic twofold axis with two C atoms of the central benzene ring lying on the axis. In the crystal, N-H⋯O hydrogen bonds link the mol-ecules into chains parallel to the c axis.

2-tert-Butyl 4-methyl 3,5-dimethyl-1H-pyrrole-2,4-dicarboxyl-ate.

In the title mol-ecule, C(13)H(19)NO(4), except for two C atoms of the tert-butyl group, the non-H atoms are almost coplanar (r.m.s. deviation = 0.2542 Å). In the crystal, mol-ecules are linked into centrosymmetric dimers by two inter-molecular N-H⋯O hydrogen bonds, forming an R(2) (2)(10) ring motif.

A simple hydrazone probe for recognition of Al3+ and PPi and its applicability in lysosomal imaging.

A simple hydrazone probe (1) was designed and synthesized for the successive detection of Al3+ and pyrophosphate (PPi) in almost 100% buffer environment. The probe provided O2N donor set for chelation with Al3+, leading to a distinct fluorescence boost at 510 nm. The in-situ formed 1-Al3+ complex detected PPi with an "on-off" behavior. The detection limits for Al3+ and PPi were 35.7 nM and 76 nM, respectively. Benefiting from the existence of morpholine as lysosome-targeting group, probe 1 was successfully applied to the detection of Al3+ and PPi in lysosomes.

Spontaneous epidural hematoma of thoracic spine presenting as Brown-Séquard syndrome: report of a case with review of the literature.

BACKGROUND: Spontaneous spinal epidural hematoma (SSEH) is an uncommon clinical entity. It produces a severe neurological deficit and prompt decompression is usually the first choice of treatment. Brown-Séquard syndrome is commonly seen in the setting of spinal trauma or an extramedullary spinal neoplasm, but rarely caused by SSEH. METHODS: Case report and literature review. FINDINGS: A previously healthy man presented with Brown-Séquard syndrome below T5-T6 cord segment secondary to spontaneous epidural hematoma. He opted for conservative treatment, which was followed by rapid resolution. CONCLUSIONS: Although Brown-Séquard syndrome as a presenting feature of SSEH is rare, it does exist in exceptional case, which should be taken into consideration for differential diagnosis. Prompt surgical decompression is an absolute surgical indication widely accepted for patient with progressive neurological deficit. However, SSEH presenting with incomplete neurological insult such as Brown-Séquard syndrome might have a benign course. Successful non-operative management of this problem does not make it a standard of care, and surgical decompression remains the standard treatment for SSEH.

3-Methyl-4-{[(3-{[(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl-idene)(phen-yl)meth-yl]amino-meth-yl}benz-yl)amino](phen-yl)methyl-idene}-1-phenyl-1H-pyrazol-5(4H)-one.

The complete mol-ecule of the title compound, C(42)H(36)N(6)O(2), is generated by a crystallographic twofold axis with two C atoms of the central phenyl group lying on the axis. In the independent part of the mol-ecule, one amino group is involved in an intra-molecular N-H⋯O hydrogen bond, and the two adjacent phenyl rings are twisted from the plane of the pyrazolone ring with dihedral angles of 6.82 (3) and 88.32 (6)°. The crystal packing exhibits no classical inter-molecular contacts.

Nontraumatic acute paraplegia associated with cervical disk herniation.

BACKGROUND: Acute paraplegia is a true emergency. It is often the result of trauma but is rarely reported in association with cervical disk herniation in patients without antecedent injury. METHODS: Case report. FINDINGS: This 75-year-old man presented with acute paraplegia due to severe compression of the spinal cord by herniation of the C4-C5 cervical disk. He underwent emergency diskectomy and anterior fusion. Postoperatively, his neurologic functions improved gradually. CONCLUSIONS: Cervical disk herniation should be considered in the differential diagnosis of nontraumatic acute paraplegia. Pre-existing narrowed canal is an important predisposing factor and excessive neck movements are believed to be triggering factors. Immediate early decompressive surgery is recommended to avoid irreversible progression of neurologic deficit.

Accuracy and Precision Evaluation of International Standard Spherical Model by Digital Dental Scanners.

With the popularization of digital technology and the exposure of traditional technology's defects, computer-aided design and computer-aided manufacturing (CAD/CAM) has been widely used in the field of dentistry. And the accuracy of the scanning system determines the ultimate accuracy of the prosthesis, which is a very important part of CAD/CAM, so we decided to evaluate the accuracy of the intraoral and extraoral scanners. In this study, we selected the sphere model as the scanning object and obtained the final result through data analysis and 3D fitting. In terms of trueness and precision, the scanner of SHINING was significantly different from that of others; however, there was no significant difference between TRIOS and CEREC. SHINING showed the lowest level of accuracy, with CEREC slightly lower than TRIOS. The sphere model has also been proven to be scanned successfully.

A novel strategy incorporated the power of mesenchymal stem cells to allografts for segmental bone tissue engineering.

Mesenchymal stem cells (MSCs) hold great promise for bone regeneration. However, the power of mesenchymal stem cells has not been applied to structural bone allografts in clinical practice. This study designed a new strategy to enhance the efficiency of allografts for segmental bone regeneration. Isolated MSCs were cultured to form a cell sheet. The MSC sheet was then wrapped onto structural allografts. The assembled structures were cultured in vitro to evaluate the differentiation potential of MSC sheet. The assembled structures were implanted subcutaneously into nude mice as well as into the segmental radius defect of rabbits to investigate the efficiency of MSC sheets to repopulate allografts for bone repair. MSC sheets, upon assembling on bone grafts, showed similar differentiation properties to the in situ periosteum in vitro. After implantation the MSC sheets accelerated the repopulation of bone grafts in nude mice. Moreover, MSC sheets induced thicker cortical bone formation and more efficient graft-to-bone end fusion at the segmental bone defects in rabbits. This study thus presented a novel, more efficient, and practical strategy for large weight-bearing bone reconstruction by using MSC sheets to deliver large number of MSCs to repopulate the bone allografts.

The influence of the pulsatility of the blood flow on the extent of platelet adhesion.

A new improved flow system was developed to study the influence of blood flow pulsatility on platelet adhesion on adhesive proteins and bio-medical materials. The pulsatility was introduced by changing the shear rate every 15 s in blood that was aspirated through a perfusion chamber by a syringe pump. The advantage of this new system is that it avoids system related platelet activation. At steady low shear rate (300/s) after 5 min a collagen type III surface was covered for 24.2+/-3.8% with platelets. At steady high shear rate (1300/s) platelet coverage to collagen was 48.8+/-6.8%. When pulsatility was introduced by changing the shear rate was every 15 s form 300/s to 1300/s and vice-versa, platelet coverage after 5 min was increased to 60.4+/-4.0% (p<0.001). After 5 min perfusion samples were taken from the perfusate and the extent of platelet activation was measured. The significant difference in surface expression of P-selectin on platelets is only seen when comparing pulse flow with control (no flow). We concluded that a significant increase in platelet activation during blood pulsatile flow compared with steady flow, which results in an increased platelet adhesion to collagen.

[Clinical observation on the effect of joint mobilization in treating elderly patients after distal radius fractures operation].

OBJECTIVE: To investigate the effect of joint mobilization on postoperative wrist joint function, pain and grip strength for elderly patients with distal radius fracture. METHODS: From January 2015 to June 2016, a total of 67 elderly patients with distal radius fracture were randomly divided into routine exercise group and joint mobilization group. Among them, 37 patients in the routine exercise group underwent conventional distal radius fracture postoperative joint function exercise regimen, including 16 males and 21 females with a mean age of (67.8±3.2) years old ranging from 60 to 72 years old;the injured side was dominant in 23 cases and non-dominant in 14 cases;injury mechanism was fall in 26 cases, traffic accident in 11 cases; for AO type, 6 cases were type B3, 18 cases were type C1, 7 cases were type C2, 6 cases was type C3. Other 30 patients in the joint mobilization group underwent joint mobilization on the basis of the routine exercise group including 14 males and 16 females with a mean age of (67.1±4.0) years old ranging from 61 to 74 years old; the injured side was dominant in 21 cases and non-dominant in 9 cases;injury mechanism was fall in 25 cases, traffic accident in 5 cases;for AO type, 8 cases were type B3, 13 cases were type C1, 6 cases were type C2, 9 cases were type C3. The wrist joint activity, Gartland-Werley wrist joint function score, VAS pain score and grip strength were observed at 3 months afrer treatment. RESULTS: After 3 months' treatment, the VAS in the routine exercise group was higher than that of the joint mobilization group (P<0.05). The grip strength of affected side in both groups were lower than that of contralateral side, but the average grip strength of affected side in joint mobilization group was higher than that in routine exercise group(P<0.05). In routine exercise group, the average angle of flexion, extension, radial deviation were significantly higher than those of joint mobilization group(P<0.05). But ulnar deviation angle in routine exercise group compared with joint mobilization group had no significant difference (P>0.05). In the comparison of each item of Gartland-Werley, there was no significant difference between two groups in residual deformity and complication(P>0.05); the average score of subjective score, objective score and total score in routine exercise group were significantly higher than those of the joint mobilization group (P<0.05). The wrist function Gartland-Werley score in routine exercise group after treatment was excellent in 21 cases, good in 10, 6 in fair, while in joint mobilization group, excellent in 23, good in 6, fair in 1(P<0.05). CONCLUSIONS: The application of joint mobilization in the treatment of elderly patients with distal radius fracture can improve the joint activity and obtain better wrist function after surgery.

Quantitative estimation of the high-intensity zone in the lumbar spine: comparison between the symptomatic and asymptomatic population.

BACKGROUND CONTEXT: The high-intensity zone (HIZ) on magnetic resonance imaging (MRI) has been studied for more than 20 years, but its diagnostic value in low back pain (LBP) is limited by the high incidence in asymptomatic subjects. Little effort has been made to improve the objective assessment of HIZ. PURPOSE: To develop quantitative measurements for HIZ and estimate intra- and interobserver reliability and to clarify different signal intensity of HIZ in patients with or without LBP. STUDY DESIGN: A measurement reliability and prospective comparative study. PATIENT SAMPLE: A consecutive series of patients with LBP between June 2010 and May 2011 (group A) and a successive series of asymptomatic controls during the same period (group B). OUTCOME MEASURES: Incidence of HIZ; quantitative measures, including area of disc, area and signal intensity of HIZ, and magnetic resonance imaging index; and intraclass correlation coefficients (ICCs) for intra- and interobserver reliability. METHODS: On the basis of HIZ criteria, a series of quantitative dimension and signal intensity measures was developed for assessing HIZ. Two experienced spine surgeons traced the region of interest twice within 4 weeks for assessment of the intra- and interobserver reliability. The quantitative variables were compared between groups A and B. RESULTS: There were 72 patients with LBP and 79 asymptomatic controls enrolling in this study. The prevalence of HIZ in group A and group B was 45.8% and 20.2%, respectively. The intraobserver agreement was excellent for the quantitative measures (ICC=0.838-0.977) as well as interobserver reliability (ICC=0.809-0.935). The mean signal of HIZ in group A was significantly brighter than in group B (57.55±14.04% vs. 45.61±7.22%, p=.000). There was no statistical difference of area of disc and HIZ between the two groups. The magnetic resonance imaging index was found to be higher in group A when compared with group B (3.94±1.71 vs. 3.06±1.50), but with a p value of .050. CONCLUSIONS: A series of quantitative measurements for HIZ was established and demonstrated excellent intra- and interobserver reliability. The signal intensity of HIZ was different in patients with or without LBP, and significant brighter signal was observed in symptomatic subjects.

Degenerative changes in the interspinous ligament.

OBJECTIVE: The aim of this study was to investigate the imaging assessment of interspinous ligament degeneration (ISLD) in patients with or without low back pain (LBP). METHODS: Sixty patients with LBP were enrolled in Group A and 60 subjects frequency-matched by age and sex in Group B. An MRI-based grading system for ISLD was scored and ranged from Type A (normal) to Type D (severe). The lumbar disc was also graded according to degeneration at four lumbar levels. RESULTS: Type A ISLD was the most prevalent type with 161 levels (67.1%) in Group A and 172 (71.7%) in Group B. Type D was the least frequent, seen in 13 levels in Group A and 3 in Group B. There was a significantly higher incidence of Type D ISLD in Group A than Group B (5.4% vs. 1.3%, p<0.05). The average age of patients with Type D ISLD in Group A was higher than Types A, B and C (Type A and B p<0.01, Type C p<0.05). In Group B, the age of patients with Type D ISLD was significantly higher than those with Type A (p<0.05). Although disc grade increased in advanced ISLD in both groups, only the difference between Type D and Types A and B in Group A were statistically significant (p<0.05). CONCLUSION: More advanced ISLD grades were less common in patients with or without LBP. Advanced change of the ISL was more common in patients with LBP. ISLD occurred in more severe disc degeneration.

Cyclic tensile strain facilitates the ossification of ligamentum flavum through ß-catenin signaling pathway: in vitro analysis.

STUDY DESIGN: Histological, immunohistochemical, and real-time reverse transcription-polymerase chain reaction analyses of the expression of cell signaling and transcriptional factors in human ossification of ligamentum flavum (OLF). OBJECTIVE: To test the hypothesis that ß-catenin plays a role in the ossification of OLF cells in response to cyclic tensile strain. SUMMARY OF BACKGROUND DATA: Several studies have investigated the roles of biomechanical and metabolic factors in the development and progression of OLF, based on the importance of genetic and biological factors. The process of ossification includes enchondral ossification, although such pathology remains poorly defined. METHODS: Using real-time reverse transcription-polymerase chain reaction, we analyzed the mRNA expression levels of signaling factors known to be involved in the ossification process (ß-catenin, Runx2, Sox9, and osteopontin) in cultured OLF cells subjected to cyclic tensile strain. Cyclic tensile strain was produced by Flexercell FX-3000 (Flexercell International, Hillsborough, NC), applied for 0, 6, 12, or 24 hours. The localization of these factors was examined in decalcified paraffin OLF sections by immunohistochemistry. Controlled samples were harvested from nonossified ligamentum flavum of patients who underwent thoracic posterior surgical procedures. RESULTS: Under resting conditions (no tensile strain), the mRNA levels of ß-catenin, Runx2, Sox9, and osteopontin in cultured OLF cells were significantly higher than in the control non-OLF cells. Application of cyclic tensile strain to OLF cells resulted in significant increases in mRNA expression levels of ß-catenin, Runx2, Sox9, and osteopontin at 24 hours. Hypertrophic chondrocytes present around the calcification front were immunopositive for Runx2 and osteopontin. Immunoreactivity of ß-catenin and Sox9 was strongly present in premature chondrocytes in the fibrocartilage area. CONCLUSION: Our results indicated that cyclic tensile strain applied to OLF cells activated their ossification through a process mediated by the ß-catenin signaling pathway.

Routinely using prophylactic antibiotic may not effectively prevent intervertebral disc infection: a new strategy to preventing postoperative intervertebral disc infection.

After spine surgery, intervertebral disc infection is a less common but disastrous complication. Since invasive techniques and procedures are increasingly used in the diagnosis and treatment of human disc diseases, the possibility of disc infection also increases. Today a large proportion of spinal operation is contributed to degenerative disc diseases (DDDs) in the elderly. With the degeneration of the disc, vertebral end plate undergoes sclerosis, and then the efficacy of antibiotics depends upon the permeability of the intervertebral disc, it is therefore likely that the penetration of the antibiotics is totally different in normal and degenerative discs. Antibiotics are often administrated prophylactically in spinal procedures to reduce the risk of infection of the disc space. However, the guideline for prophylactic use of antibiotics does not take into account the unique environment of intervertebral disc and the changes occurred in degenerative disc. Routinely using prophylactic antibiotic may not result in adequate antibiotic concentration in intervertebral disc to prevent disc infection, especially in DDD patients. We hypothesized that during the administration of antibiotic, prolonged lying flat on bed could decrease the internal pressure of the disc, and then increase the penetration of antibiotic into disc. According to different stages of end-plate sclerosis on MRI, combination of lying flat and increasing duration and concentration of antibiotics may be a new strategy to prevent postoperative intervertebral disc infection.

Ossification process involving the human thoracic ligamentum flavum: role of transcription factors.

INTRODUCTION: Ossification of the ligamentum flavum (OLF) of the spine is associated with serious neurologic compromise, but the pathomechanism of this process remains unclear. The objective of this study was to investigate the pathomechanism of the ossification process, including the roles of various transcriptional factors in the ossification of human thoracic ligamentum flavum. METHODS: Sections of the thoracic ligamentum flavum were obtained from 31 patients with OLF who underwent posterior thoracic decompression, and from six control patients free of OLF. Cultured ligamentum flavum cells (n = 6, each) were examined with real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis for Sry-type high-mobility group box 9 (Sox9), runt-related transcription factor 2 (Runx2), muscle segment homeobox 2 (Msx2), Osterix, distal-less homeobox 5 (Dlx5), and AP-1. The harvested sections were examined with hematoxylin-eosin, the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method, and immunohistochemistry for the transcriptional factors. RESULTS: Compared with the control, the OLF showed disorganization of the elastic fiber bundles and abundant hypertrophic chondrocytes in the ossification front. TUNEL-positive chondrocytes were found near the ossified plaques. The mRNA expression levels of Sox9, Runx2, Msx2, and AP-1 in cultured cells from the ligamentum flavum of OLF patients were significantly different from those of the control. OLF samples were strongly immunoreactive to Sox9, Runx2, and Msx2 at proliferating chondrocytes in the fibrocartilage area. Hypertrophic chondrocytes were positive for Runx2, Osterix, Dlx5, and AP-1. CONCLUSIONS: The ossification process in OLF seems to involve chondrocyte differentiation under the unique expression of transcriptional factors. Accumulation of hypertrophic chondrocytes was evident around the calcified area at the ossification front, and we suggest that the differentiation of these cells seems to be concerned with the ossification process.

Electrospun nanofibrous matrix improves the regeneration of dense cortical bone.

Numerous in vitro studies have indicated the potential of using electrospun nanofibrous scaffolds for tissue regeneration. However, few reports have demonstrated their utility in real tissue repair models. The present investigation tested the hypothesis that electrospun poly-L-lactic acid (PLLA) nanofibrous membrane leads to dense cortical bone regeneration and improves the efficacy of currently-used collagenous guided bone regeneration (GBR) membrane. In vitro, the function of bone marrow-derived mesenchymal stem cells (BMSCs) on nanofibrous scaffolds was evaluated. In an in vivo experiment, large bony defects were created in rabbit tibia and treated with a nanofiber-reinforced bilayer membrane, nanofibrous membrane, or collagenous membrane alone. Three and six weeks after operation, bone defect healing was assessed radiologically and histologically. In vitro differentiation studies showed that BMSCs had much higher expression of Runx2 and collagen type I, alpha 1 mRNAs, when cultured on nanofibrous scaffolds. The radiographic and histological data both showed that the group treated with bilayer membrane had more bony tissue formation at 3 weeks. Moreover, at 6 weeks, only the bilayer membrane-treated bone defects displayed better regeneration of cortical bone tissue, whereas in the other groups the defects were filled with spongy bone-like tissue. The results demonstrated that electrospun nanofibrous membrane improves the regeneration of cortical bone, suggesting that this type of membrane can be combined with current collagenous GBR membrane to improve guided bone regeneration technology.