Filamin C is Essential for mammalian myocardial integrity.

FLNC, encoding filamin C, is one of the most mutated genes in dilated and hypertrophic cardiomyopathy. However, the precise role of filamin C in mammalian heart remains unclear. In this study, we demonstrated Flnc global (FlncgKO) and cardiomyocyte-specific knockout (FlnccKO) mice died in utero from severely ruptured ventricular myocardium, indicating filamin C is required to maintain the structural integrity of myocardium in the mammalian heart. Contrary to the common belief that filamin C acts as an integrin inactivator, we observed attenuated activation of ß1 integrin specifically in the myocardium of FlncgKO mice. Although deleting ß1 integrin from cardiomyocytes did not recapitulate the heart rupture phenotype in Flnc knockout mice, deleting both ß1 integrin and filamin C from cardiomyocytes resulted in much more severe heart ruptures than deleting filamin C alone. Our results demonstrated that filamin C works in concert with ß1 integrin to maintain the structural integrity of myocardium during mammalian heart development.

CD248-expressing cancer-associated fibroblasts induce non-small cell lung cancer metastasis via Hippo pathway-mediated extracellular matrix stiffness.

Metastasis is a crucial stage in tumour progression, and cancer-associated fibroblasts (CAFs) support metastasis through their participation in extracellular matrix (ECM) stiffness. CD248 is a possible biomarker for non-small cell lung cancer (NSCLC)-derived CAFs, but its role in mediating ECM stiffness to promote NSCLC metastasis is unknown. We investigated the significance of CD248+ CAFs in activating the Hippo axis and promoting connective tissue growth factor (CTGF) expression, which affects the stromal collagen I environment and improves ECM stiffness, thereby facilitating NSCLC metastasis. In this study, we found that higher levels of CD248 in CAFs induced the formation of collagen I, which in turn increased extracellular matrix stiffness, thereby enabling NSCLC cell infiltration and migration. Hippo axis activation by CD248+ CAFs induces CTGF expression, which facilitates the formation of the collagen I milieu in the stromal matrix. In a tumour lung metastasis model utilizing fibroblast-specific CD248 gene knockout mice, CD248 gene knockout mice showed a significantly reduced ability to develop tumour lung metastasis compared to that of WT mice. Our findings demonstrate that CD248+ CAFs activate the Hippo pathway, thereby inducing CTGF expression, which in turn facilitates the collagen I milieu of the stromal matrix, which promotes NSCLC metastasis.

Metformin induces tolerogenicity of dendritic cells by promoting metabolic reprogramming.

Dendritic cells (DCs) can mediate immune responses or immune tolerance depending on their immunophenotype and functional status. Remodeling of DCs' immune functions can develop proper therapeutic regimens for different immune-mediated diseases. In the immunopathology of autoimmune diseases (ADs), activated DCs notably promote effector T-cell polarization and exacerbate the disease. Recent evidence indicates that metformin can attenuate the clinical symptoms of ADs due to its anti-inflammatory properties. Whether and how the therapeutic effects of metformin on ADs are associated with DCs remain unknown. In this study, metformin was added to a culture system of LPS-induced DC maturation. The results revealed that metformin shifted DC into a tolerant phenotype, resulting in reduced surface expression of MHC-II, costimulatory molecules and CCR7, decreased levels of proinflammatory cytokines (TNF-α and IFN-γ), increased level of IL-10, upregulated immunomodulatory molecules (ICOSL and PD-L) and an enhanced capacity to promote regulatory T-cell (Treg) differentiation. Further results demonstrated that the anti-inflammatory effects of metformin in vivo were closely related to remodeling the immunophenotype of DCs. Mechanistically, metformin could mediate the metabolic reprogramming of DCs through FoxO3a signaling pathways, including disturbing the balance of fatty acid synthesis (FAS) and fatty acid oxidation (FAO), increasing glycolysis but inhibiting the tricarboxylic acid cycle (TAC) and pentose phosphate pathway (PPP), which resulted in the accumulation of fatty acids (FAs) and lactic acid, as well as low anabolism in DCs. Our findings indicated that metformin could induce tolerance in DCs by reprogramming their metabolic patterns and play anti-inflammatory roles in vitro and in vivo.

An in-library ligation strategy and its application in CRISPR/Cas9 screening of high-order gRNA combinations.

Simultaneous targeting multiple genes is a big advantage of CRISPR (clustered regularly interspaced short palindromic repeats) genome editing but challenging to achieve in CRISPR screening. The crosstalk among genes or gene products is a common and fundamental mechanism to ensure cellular stability and functional diversity. However, the screening approach to map high-order gene combinations to the interesting phenotype is still lacking. Here, we developed a universal in-library ligation strategy and applied it to generate multiplexed CRISPR library, which could perturb four pre-designed targets in a cell. We conducted in vivo CRISPR screening for potential guide RNA (gRNA) combinations inducing anti-tumor immune responses. Simultaneously disturbing a combination of three checkpoints in CD8+ T cells was demonstrated to be more effective than disturbing Pdcd1 only for T cell activation in the tumor environment. This study developed a novel in-library ligation strategy to facilitate the multiplexed CRISPR screening, which could extend our ability to explore the combinatorial outcomes from coordinated gene behaviors.

Primary Mouse Aortic Smooth Muscle Cells Exhibit Region- and Sex-Dependent Biological Responses In Vitro.

Despite advancements in elucidating biological mechanisms of cardiovascular remodeling, cardiovascular disease (CVD) remains the leading cause of death worldwide. When stratified by sex, clear differences in CVD prevalence and mortality between males and females emerge. Regional differences in phenotype and biological response of cardiovascular cells are important for localizing the initiation and progression of CVD. Thus, to better understand region and sex differences in CVD presentation, we have focused on characterizing in vitro behaviors of primary vascular smooth muscle cells (VSMCs) from the thoracic and abdominal aorta of male and female mice. VSMC contractility was assessed by traction force microscopy (TFM; single cell) and collagen gel contraction (collective) with and without stimulation by transforming growth factor-beta 1 (TGF-ß1) and cell proliferation was assessed by a colorimetric metabolic assay (MTT). Gene expression and TFM analysis revealed region- and sex-dependent behaviors, whereas collagen gel contraction was consistent across sex and aortic region under baseline conditions. Thoracic VSMCs showed a sex-dependent sensitivity to TGF-ß1-induced collagen gel contraction (female > male; p = 0.025) and a sex-dependent proliferative response (female > male; p < 0.001) that was not apparent in abdominal VSMCs. Although primary VSMCs exhibit intrinsic region and sex differences in biological responses that may be relevant for CVD presentation, several factors-such as inflammation and sex hormones-were not included in this study. Such factors should be included in future studies of in vitro mechanobiological responses relevant to CVD differences in males and females.

Validating the knowledge bank approach for personalized prediction of survival in acute myeloid leukemia: a reproducibility study.

Reproducibility is not only essential for the integrity of scientific research but is also a prerequisite for model validation and refinement for the future application of predictive algorithms. However, reproducible research is becoming increasingly challenging, particularly in high-dimensional genomic data analyses with complex statistical or algorithmic techniques. Given that there are no mandatory requirements in most biomedical and statistical journals to provide the original data, analytical source code, or other relevant materials for publication, accessibility to these supplements naturally suggests a greater credibility of the published work. In this study, we performed a reproducibility assessment of the notable paper by Gerstung et al. (Nat Genet 49:332-340, 2017) by rerunning the analysis using their original code and data, which are publicly accessible. Despite an open science setting, it was challenging to reproduce the entire research project; reasons included: incomplete data and documentation, suboptimal code readability, coding errors, limited portability of intensive computing performed on a specific platform, and an R computing environment that could no longer be re-established. We learn that the availability of code and data does not guarantee transparency and reproducibility of a study; paradoxically, the source code is still liable to error and obsolescence, essentially due to methodological and computational complexity, a lack of reproducibility checking at submission, and updates for software and operating environment. The complex code may also hide problematic methodological aspects of the proposed research. Building on the experience gained, we discuss the best programming and software engineering practices that could have been employed to improve reproducibility, and propose practical criteria for the conduct and reporting of reproducibility studies for future researchers.

Efficacy of using tidal volume challenge to improve the reliability of pulse pressure variation reduced in low tidal volume ventilated critically ill patients with decreased respiratory system compliance.

BACKGROUND: The prediction accuracy of pulse pressure variation (PPV) for fluid responsiveness was proposed to be unreliable in low tidal volume (Vt) ventilation. It was suggested that changes in PPV obtained by transiently increasing Vt to 8 ml/kg accurately predicted fluid responsiveness even in subjects receiving low Vt. We assessed whether the changes in PPV induced by a Vt challenge predicted fluid responsiveness in our critically ill subjects ventilated with low Vt 6 ml/kg. METHODS: This study is a prospective single-center study. PPV and other parameters were measured at a Vt of 6 mL/kg, 8 mL/kg, and after volume expansion. The prediction accuracy of PPV and other parameters for fluid responsiveness before and after tidal volume challenge was also analyzed using receiver operating characteristic (ROC) curves. RESULTS: Thirty-one of the 76 subjects enrolled in the study were responders (41%). Respiratory system compliance of all subjects decreased significantly (26 ± 4.3). The PPV values were significantly higher in the responder group than the non-responder group before (8.8 ± 2.7 vs 6.8 ± 3.1) or after (13.0 ± 1.7 vs 8.5 ± 3.0) Vt challenge. In the receiver operating characteristic curve (ROC) analysis, PPV6 showed unsatisfactory predictive capability with an area under the curve (AUC) of 0.69 (95%CI, 0.57-0.79, p = 0.002) at a Vt of 6 mL/kg. PPV8 andΔPPV6-8 showed good predictive capability with an AUC of 0.90 (95% CI, 0.81-0.96, p < 0.001) and 0.90 (95% CI, 0.80-0.95, P < 0.001) respectively. The corresponding cutoff values were 11% for PPV8 and 2% for ΔPPV6-8. CONCLUSIONS: PPV shows a poor operative performance as a predictor of fluid responsiveness in critically ill subjects ventilated with a tidal volume of 6 mL/ kg. Vt challenge could improve the predictive accuracy of PPV to a good but not excellent extent when respiratory system compliance decreased significantly.

Ferumoxytol-ß-glucan Inhibits Melanoma Growth via Interacting with Dectin-1 to Polarize Macrophages into M1 Phenotype.

Background: Regulating the polarization of macrophages to antitumor M1 macrophages is a promising strategy for overcoming the immunosuppression of the tumor microenvironment for cancer therapy. Ferumoxytol (FMT) can not only serve as a drug deliver agent but also exerts anti-tumor activity. ß-glucan has immuno-modulating properties to prevent tumor growth. Thus, a nanocomposite of FMT surface-coated with ß-glucan (FMT-ß-glucan) was prepared to explore its effect on tumor suppression. Methods: Male B16F10 melanoma mouse model was established to explore the antitumor effect of FMT-ß-glucan. The viability and apoptotic rates of B16F10 cells were detected by cell counting kit-8 and Annexin-V/PI experiments. The levels of M1 markers were quantified by quantitative reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay. Phagocytic activity and intracellular reactive oxygen species (ROS) in macrophages were evaluated by the neutral red uptake assay and flow cytometry, respectively. Small interfering RNA (siRNA) transfection was applied to knock down the Dectin-1 gene in RAW 264.7 cells. Results: FMT-ß-glucan suppressed tumor growth to a greater extent and induced higher infiltration of M1 macrophages than the combination of FMT and ß-glucan (FMT+ß-glucan) in vivo. In vitro, supernatant from FMT-ß-glucan-treated RAW 264.7 cells led to lower cell viability and induced more apoptosis of B16F10 cells than that from the FMT+ß-glucan group. Moreover, FMT-ß-glucan boosted the expression of M1 type markers, and increased phagocytic activity and ROS in RAW 264.7 cells. Further research indicated that FMT-ß-glucan treatment promoted the level of Dectin-1 on the surface of RAW 264.7 cells and that knockdown of Dectin-1 abrogated the phosphorylation levels of several components in MAPK and NF-κB signaling. Conclusion: The nanocomposite FMT-ß-glucan suppressed melanoma growth by inducing the M1 macrophage-activated tumor microenvironment.

Efficacy of High-Flow Nasal Cannula Oxygen Therapy in Patients with Mild Hypercapnia.

OBJECTIVES: To investigate the indications of high-flow nasal cannula (HFNC) oxygen therapy among patients with mild hypercapnia and to explore the predictors of intubation when HFNC fails. METHODS: This retrospective study was conducted based on the Medical Information Mart for Intensive Care IV (MIMIC-IV) database. Adult patients with mild hypercapnia (45 < PaCO2 ≤ 60 mmHg) received either HFNC or non-invasive ventilation (NIV) oxygen therapy. Propensity score matching (PSM) was implemented to increase between-group comparability. The Kaplan-Meier method was used to estimate overall survival and cumulative intubation rates, while 28-day mortality and 48-h and 28-day intubation rates were compared using the Chi-squared test. The predictive performances of HR/SpO2 and the ROX index (the ratio of SpO2/FiO2 to respiratory rate) at 4 h were assessed regarding HFNC failure, which was determined if intubation was given within 48 h after the initiation of oxygen therapy. The area under the receiver operating characteristic curve (AUC) for HR/SpO2 and the ROX index were calculated and compared. RESULTS: A total of 524,520 inpatient hospitalization records were screened, 106 patients in HFNC group and 106 patients in NIV group were successfully matched. No significant difference in 48-h intubation rate between the HFNC group (the treatment group) and the NIV group (the control group) (14.2% vs. 8.5%, p = 0.278); patients receiving HFNC had higher 28-day intubation rate (26.4% vs. 14.2%, p = 0.029), higher 28-day mortality (17.9% vs. 8.5%, p = 0.043), and longer ICU length of stay (4.4 vs. 3.3 days, p = 0.019), compared to those of NIV group. The AUC of HR/SpO2 at 4 h after the initiation of HFNC yielded around 0.660 for predicting 48-h intubation, greater than that of the ROX index with an AUC of 0.589 (p < 0.01). CONCLUSION: Patients with impending respiratory failure had lower intubation rate, shorter ICU length of stay, and lower mortality when treated mild hypercapnia with NIV over HFNC. As opposed to the ROX index, a modest, yet improved predictive performance is demonstrated using HR/SpO2 in predicting the failure of HFNC among these patients.

Real-time MR-guided brain biopsy using 1.0-T open MRI scanner.

OBJECTIVES: To evaluate the safety, feasibility and diagnostic performance of real-time MR-guided brain biopsy using a 1.0-T open MRI scanner. METHODS: Medical records of 86 consecutive participants who underwent brain biopsy under the guidance of a 1.0-T open MRI scanner with real-time and MR fluoroscopy techniques were evaluated retrospectively. All procedures were performed under local anaesthesia and intravenous conscious sedation. Diagnostic yield, diagnostic accuracy, complication rate and procedure duration were assessed. The lesions were divided into two groups according to maximum diameters: ≤ 1.5 cm (n = 16) and > 1.5 cm (n = 70). The two groups were compared using Fisher's exact test. RESULTS: Diagnostic yield and diagnostic accuracy were 95.3% and 94.2%, respectively. The diagnostic yield of lesions ≤ 1.5 cm and > 1.5 cm were 93.8% and 95.7%, respectively. There was no significant difference in diagnostic yield between the two groups (p > 0.05). Mean procedure duration was 41 ± 5 min (range 33-49 min). All biopsy needles were placed with one pass. Complication rate was 3.5% (3/86). Minor complications included three cases of a small amount of haemorrhage. No serious complications were observed. CONCLUSIONS: Real-time MR-guided brain biopsy using a 1.0-T open MRI scanner is a safe, feasible and accurate diagnostic technique for pathological diagnosis of brain lesions. The procedure duration is shortened and biopsy work flow is simplified. It could be considered as an alternative for brain biopsy. KEY POINTS: • Real-time MRI-guided brain biopsy using a 1.0-T open MRI scanner is safe, feasible and accurate. • No serious complications occurred in real-time MRI-guided brain biopsy. • Procedure duration is shortened and biopsy work flow is simplified.

Predictive role of estimated glomerular filtration rate prior to surgery in postsurgical acute kidney injury among very elderly patients: a retrospective cohort study.

Background: Acute kidney injury (AKI) is a common complication after surgery. Because of unpredictable and variable age-dependent physical decline, the incidence, risk factor of postsurgical AKI and the predictive power of estimated glomerular filtration rate prior to surgery (eGFRpreSurg) has not been fully elucidated in very elderly patients. Methods: All discharged patients aged ≥80 years without chronic kidney disease who underwent surgery prior to intensive care unit (ICU) admission from 2017 through 2018 were included. Clinical, biological and surgical data were recorded. Mean of outpatient creatinine values from the year prior to ICU admission was used as the baseline value to determine the occurrence of AKI. Postoperative AKI was diagnosed according to Kidney Disease Improving Global Outcomes criteria. Results: Among 243 very elderly postoperative patients admitted during the study period, 48 had AKI during their ICU stay. The occurrence of postsurgical AKI was associated with longer ventilation times (p < .001) and higher mortality (p < .001). The eGFRpreSurg, which is calculated based on the Modification of Diet in Renal Disease study equation, was a risk factor for postoperative AKI (OR = 2.662, p = .010). The incidence of postoperative AKI was significantly higher among patients with lower eGFRpreSurg than among those with an eGFRpreSurg ≥ 70 mL/min/1.73 m2 (p = .003). Conclusion: Postsurgical AKI in very elderly patients has a high incidence and is a risk factor for mortality. Our study confirmed that eGFRpreSurg could be used as an index for AKI risk stratification.

Long non-coding RNA LINC00968 acts as oncogene in NSCLC by activating the Wnt signaling pathway.

Long non-coding RNAs (lncRNAs) have played critical roles in a variety of cancers, including non-small cell lung cancer (N SCLC). In our study, we focused on the biological function and clinical significance of lncRNA LINC00968 in NSCLC. It was indicated that LINC00968 was significantly increased in LUAD tissues, LUSC tissues and NSCLC cells compared to their corresponding controls. Inhibition of LINC00968 was able to repress NSCLC growth, migration, and invasion in vitro while upregulation of LINC00968 reversed this process. Additionally, downregulation of LINC00968 induced apoptosis capacity of A549 cell. Apoptosis-related proteins BCL-2 were decreased and BAX was increased by knockdown of LINC00968, respectively. Meanwhile we observed that Wnt signaling pathway was involved in the LINC00968-induced NSCLC progression. Finally, in vivo tumor xenografts were established using A549 cells to detect the function of LINC00968 in NSCLC tumorigenesis. Silencing LINC00968 greatly inhibited NSCLC tumor progression, which was consistent with the in vitro tests. In conclusion, we have uncovered that LINC00968 could be regarded as a novel prognostic biomarker and therapeutic target in NSCLC diagnosis and treatment.

1.0T MR-guided percutaneous coaxial cutting needle biopsy in pancreatic lesion diagnosis.

BACKGROUND: Pancreatic carcinoma is a common cause of cancer deaths worldwide. Biopsy is often required for the initial diagnosis of pancreatic masses. Biopsy can be performed endoscopically or percutaneously with computed tomography (CT) and ultrasound (US) guidance. MRI offers many inherent advantages over CT and US. PURPOSE/HYPOTHESIS: To prospectively evaluate the feasibility, accuracy, and safety of MRI-guided percutaneous coaxial cutting needle biopsy of pancreatic lesions using an open 1.0T high-field MR scanner. STUDY TYPE: Prospective. POPULATION: Thirty-one patients with 31 pancreatic lesions underwent MR-guided percutaneous coaxial cutting needle biopsy. FIELD STRENGTH/SEQUENCE: 1.0T T2 WI-TSE PDW-aTSE T1 WI-TFE. ASSESSMENT: Final diagnosis was confirmed by surgery and clinical follow-up for at least 12 months. The accuracy, sensitivity, and specificity were calculated. Complications were recorded. STATISTICAL TESTS: There was no statistical analysis in this study. RESULTS: The procedure was technically successful and final biopsy samples were adequate for histopathological examination in all patients. Biopsy pathology revealed malignant pancreatic tumor in 25 patients (25/31, 80.6%), and benign pancreatic lesions were present in six patients (6/31, 19.4%). The final diagnosis was pancreatic malignancy in 27 patients and benign disease in four patients, which was confirmed by surgery and clinical follow-up. Two biopsy results were false-negative. The diagnostic accuracy in biopsies was 93.5% (29 of 31). The sensitivity to detect a malignant disease was 92.6% (25 of 27), and the specificity was 100%. All patients tolerated the procedure well; minor peripancreatic hemorrhage was found in two patients after the procedure, and none had major complications either during or after the procedure. DATA CONCLUSION: MRI-guided percutaneous biopsy of pancreatic lesions using an open 1.0T high-field scanner has high diagnostic accuracy, which is feasible and safe for use in clinical practice. LEVEL OF EVIDENCE: 2 Technical Efficacy: Stage 2 J. MAGN. RESON. IMAGING 2018;48:382-388.

MR-guided percutaneous biopsy of solitary pulmonary lesions using a 1.0-T open high-field MRI scanner with respiratory gating.

OBJECTIVES: To prospectively evaluate the feasibility, safety and accuracy of MR-guided percutaneous biopsy of solitary pulmonary lesions using a 1.0-T open MR scanner with respiratory gating. METHODS: Sixty-five patients with 65 solitary pulmonary lesions underwent MR-guided percutaneous coaxial cutting needle biopsy using a 1.0-T open MR scanner with respiratory gating. Lesions were divided into two groups according to maximum lesion diameters: ≤2.0 cm (n = 31) and >2.0 cm (n = 34). The final diagnosis was established in surgery and subsequent histology. Diagnostic accuracy, sensitivity and specificity were compared between the groups using Fisher's exact test. RESULTS: Accuracy, sensitivity and specificity of MRI-guided percutaneous pulmonary biopsy in diagnosing malignancy were 96.9 %, 96.4 % and 100 %, respectively. Accuracy, sensitivity and specificity were 96.8 %, 96.3 % and 100 % for lesions 2.0 cm or smaller and 97.1 %, 96.4 % and 100 %, respectively, for lesions larger than 2.0 cm. There was no significant difference between the two groups (P > 0.05). Biopsy-induced complications encountered were pneumothorax in 12.3 % (8/65) and haemoptysis in 4.6 % (3/65). There were no serious complications. CONCLUSIONS: MRI-guided percutaneous biopsy using a 1.0-T open MR scanner with respiratory gating is an accurate and safe diagnostic technique in evaluation of pulmonary lesions. KEY POINTS: • MRI-guided percutaneous lung biopsy using a 1.0-T open MR scanner is feasibility. • 96.9 % differentiation accuracy of malignant and benign lung lesions is possible. • No serious complications occurred in MRI-guided lung biopsy.

Extremely low frequency magnetic fields regulate differentiation of regulatory T cells: Potential role for ROS-mediated inhibition on AKT.

Our previous studies showed that extremely low frequency magnetic fields (ELF-MFs) inhibited tumor growth and change proportion of splenic regulatory T cells (Treg cells). Here, we focus on the effect of ELF-MFs on lung metastatic melanoma mouse model and the regulatory mechanism of ELF-MFs on the differentiation of Treg cells. Tumor-bearing mice were exposed to sham ELF-MFs and ELF-MFs (0.4 T, 7.5 Hz) 2 h/day for 27 days. Metastatic tumor burden of lung was significantly decreased after ELF-MF treatment. Compared to the control group, expressions of matrix metalloproteinase (MMP2, MMP9) and forkhead box P3 (Foxp3) in lung nodules significantly decreased in the ELF-MF group. Moreover, in vitro, after stimulated with anti-CD3, anti-CD28 antibodies and transforming growth factor-ß (TGF-ß) and treated with ELF-MFs for 2 h, expression of Foxp3 in total T cells was significantly decreased. Differentiation rate of Treg cells was inhibited from 32.0% to 22.1% by ELF-MFs. Furthermore, reactive oxygen species (ROS) was increased and phospho-serine/threonine protein kinase (p-AKT) was inhibited in both T cells and Jurkat cells. ROS scavenger N-acetyl-l-cysteine reversed inhibition of AKT pathway and expression of Foxp3 from 18.6% to 26.6% in T cells. Taken together, our data show that ELF-MF exposure promoted the inhibitory effect of ROS on AKT pathway and decreased Foxp3 expression, which provides an explanation for why ELF-MF exposure can inhibit differentiation of Treg cells and enhance antitumor effect in metastatic melanoma mouse model.

Mutant LRP6 Impairs Endothelial Cell Functions Associated with Familial Normolipidemic Coronary Artery Disease.

Mutations in the genes low-density lipoprotein (LDL) receptor-related protein-6 (LRP6) and myocyte enhancer factor 2A (MEF2A) were reported in families with coronary artery disease (CAD). We intend to determine the mutational spectrum of these genes among hyperlipidemic and normolipidemic CAD families. Forty probands with early-onset CAD were recruited from 19 hyperlipidemic and 21 normolipidemic Chinese families. We sequenced all exons and intron-exon boundaries of LRP6 and MEF2A, and found a novel heterozygous variant in LRP6 from a proband with normolipidemic CAD. This variant led to a substitution of histidine to tyrosine (Y418H) in an evolutionarily conserved domain YWTD in exon 6 and was not found in 1025 unrelated healthy individuals. Co-segregated with CAD in the affected family, LRP6Y418H significantly debilitated the Wnt3a-associated signaling pathway, suppressed endothelial cell proliferation and migration, and decreased anti-apoptotic ability. However, it exhibited no influences on low-density lipoprotein cholesterol uptake. Thus, mutation Y418H in LRP6 likely contributes to normolipidemic familial CAD via impairing endothelial cell functions and weakening the Wnt3a signaling pathway.

Serum IL-17F combined with VEGF as potential diagnostic biomarkers for oral squamous cell carcinoma.

Although interleukin (IL) 17A can promote angiogenesis in several tumors, there are limited clinical evidences on cancer about the correlation between serum vascular endothelial growth factor (VEGF) and IL-17F, which is the most homologous to IL-17A. In this study, serum concentration of IL-17F and VEGF from healthy (n = 28), leukoplakia (n = 15), and oral squamous cell carcinoma (OSCC) groups (n = 85) were assessed and showed that IL-17F level was remarkably downregulated from healthy group (394.3 pg/ml) to OSCC group (82.96 pg/ml). Conversely, the OSCC group had a highest level of VEGF (P < 0.05) in whole groups, and there was a negative correlation between IL-17F and VEGF in serum or in the peripheral blood mononuclear cells (PBMCs) at mRNA level. Moreover, the lowest ratio of IL-17F/VEGF was found in OSCC patients (P < 0.05) and lower ratio of IL-17F/VEGF correlated to higher tumor stage and lymph node metastasis. Furthermore, the serum level of IL-17F and the ratio of IL-17F/VEGF were positively associated with the numbers of CD3(+) CD4(+) T cells, which indicated that serum IL-17F could originate from PBMCs during the development of OSCC, and could be used for the diagnosis by effectively distinguishing OSCC patients from healthy individuals.

MRI-guided percutaneous transpedicular biopsy of thoracic and lumbar spine using a 0.23t scanner with optical instrument tracking.

PURPOSE: To prospectively evaluate the safety and accuracy of magnetic resonance imaging (MRI)-guided percutaneous transpedicular biopsy of thoracic and lumbar spine using 0.23T magnetic resonance imaging with optical tracking. MATERIALS AND METHODS: Sixty-seven thoracic and lumbar spine lesions in 67 patients underwent MRI-guided percutaneous transpedicular biopsy using 0.23T MRI with optical tracking. These lesions were divided into two groups according to the location: 16 lesions in the thoracic spine and 51 lesions in the lumbar spine. The diagnostic accuracy, sensitivity, and specificity were calculated, and comparison of the two groups was performed using Fisher's exact test. Each patient was monitored for complications. RESULTS: All specimens obtained were sufficient for diagnosis. Histological examination of MRI-guided percutaneous biopsy revealed 38 malignant and 29 benign lesions. The final diagnoses from surgery or clinical follow-up were 42 malignant lesions and 25 benign lesions. The combined diagnostic performance of MRI-guided percutaneous transpedicular thoracic and lumbar biopsy in diagnosing malignant tumors was as follows: accuracy, 94%; sensitivity, 89%; and specificity, 100%. There was no significant difference between the two groups (P = 1, Fisher's exact test). No serious complications occurred. CONCLUSION: MRI-guided percutaneous transpedicular biopsy is a safe and accurate diagnostic technique to evaluate thoracic and lumbar spine lesions.

Gene delivery with IFN-γ-expression plasmids enhances the therapeutic effects of MSCs on DSS-induced mouse colitis.

OBJECTIVE: Interferon-γ (IFN-γ) is known to enhance the immunosuppressive properties of mesenchymal stem cells (MSCs). The aim of this study was to determine whether gene modification with IFN-γ-expression plasmids could boost the therapeutic effects of MSCs on DSS-induced colitis. METHODS: We first reconstructed pcDNA3.1-IFNγ plasmids, transfected them to human umbilical cord derived MSCs, and detected the basic characters of MSCs including immune phenotype, cell vitality, proliferation, apoptosis and cell cycle progression after transfection. Subsequently, we analyzed the inhibition effect of IFN-γ-MSCs on T cell proliferation in vitro. Finally, we induced colitis in female C57BL/6 mice by 3 % DSS treatment and evaluated the therapeutic efficacy of IFN-γ-MSCs on colitis. RESULTS: Transfection with pcDNA3.1-IFNγ did not change the basic characters of MSCs. Interestingly, IFN-γ-MSCs showed more potent immunosuppressive effects on the proliferation of T cells compared to normal MSCs. Furthermore, systemic infusion with IFN-γ-MSCs more efficiently ameliorated DSS-induced mouse colitis including colitis-related ease of body weight, increase of colon length, decrease of disease activity index, and improvement of small intestine tissues structure. In addition, IFN-γ-MSCs increased the populations of Foxp3(+) Tregs and Th2 cells both in mesenteric lymph node and spleen, upregulated indoleamine 2, 3-dioxygenase expression, and suppressed inflammatory cytokine production in mouse colon. CONCLUSIONS: Gene delivery with IFN-γ-expression plasmids enhanced the therapeutic effects of MSCs on DSS-induced mouse colitis. This study provides an effective therapeutic strategy of MSCs for inflammatory diseases.

Boosting Oxygen Reduction at Pt(111)|Proton Exchange Ionomer Interfaces through Tuning the Microenvironment Water Activity.

A proton exchange ionomer is one of the most important components in membrane electrode assemblies (MEAs) of polymer electrolyte membrane fuel cells (PEMFCs). It acts as both a proton conductor and a binder for nanocatalysts and carbon supports. The structure and the wetting conditions of the MEAs have a great impact on the microenvironment at the three-phase interphases in the MEAs, which can significantly influence the electrode kinetics such as the oxygen reduction reaction (ORR) at the cathode. Herein, by using the Pt(111)|X ionomer interface as a model system (X = Nafion, Aciplex, D72), we find that higher drying temperature lowers the onset potential for sulfonate adsorption and reduces apparent ORR current, while the current wave for OHad formation drops and shifts positively. Surprisingly, the intrinsic ORR activity is higher after properly correcting the blocking effect of Pt active sites by sulfonate adsorption and the poly(tetrafluoroethylene) (PTFE) skeleton. These results are well explained by the reduced water activity at the interfaces induced by the ionomer/PTFE, according to the mixed potential effect. Implications for how to prepare MEAs with improved ORR activity are provided.