Preoperative hs-CRP/HDL ratio is associated with increased risk for postoperative SIRS in elderly patients: a retrospective cohort study.

BACKGROUND: Systemic inflammatory response syndrome (SIRS) greatly affects postoperative lives of afflicted aged patients. This study aimed to determine whether preoperative high hs-CRP/HDL ratio (CHR) was associated with an increased risk of postoperative SIRS in the elderly population. METHODS: This retrospective cohort study included data on patients aged ≥ 65 years who underwent general anesthesia surgery at two clinical centers between January 2015 and September 2020. The primary exposure was preoperative CHR which was divided into two groups (≤ 12.82 and > 12.82) based on its normal range in our hospital, and the primary outcome was the incidence of postoperative SIRS. Targeted maximum likelihood estimation analyses were used to model the exposure-outcome relationship. RESULTS: The analysis included 5595 elderly patients, of whom 1410 (25.20%) developed SIRS within three postoperative days. Targeted maximum likelihood estimation analysis revealed that elderly patients with CHR > 12.82 vs. CHR ≤ 12.82 was associated with increased risk of postoperative SIRS (aOR = 1.40, 95% CI [1.33, 1.48], P < 0.001). Those results were consistent both in subgroup analyses and sensitivity analyses. Compared with patients with CHR ≤ 12.82, patients with CHR > 12.82 had a higher prevalence of postoperative SIRS (49.06% vs. 22.70%), postoperative in-hospital mortality (3.40% vs. 0.65%), a longer hospital stay after surgery [10 (IQR, 6-16) vs. 8 (IQR, 5-11) days] and higher direct medical cost [10070 (IQR, 6878-15577) vs. 7117 (IQR, 4079-10314) euros, all P < 0.001]. CONCLUSIONS: In elderly patients, preoperative CHR > 12.82 was significantly associated with a higher risk of postoperative SIRS.

Development and validation of a machine learning model to predict prognosis in HIV-negative cryptococcal meningitis patients: a multicenter study.

PURPOSE: To predict prognosis in HIV-negative cryptococcal meningitis (CM) patients by developing and validating a machine learning (ML) model. METHODS: This study involved 523 HIV-negative CM patients diagnosed between January 1, 1998, and August 31, 2022, by neurologists from 3 tertiary Chinese centers. Prognosis was evaluated at 10 weeks after the initiation of antifungal therapy. RESULTS: The final prediction model for HIV-negative CM patients comprised 8 variables: Cerebrospinal fluid (CSF) cryptococcal count, CSF white blood cell (WBC), altered mental status, hearing impairment, CSF chloride levels, CSF opening pressure (OP), aspartate aminotransferase levels at admission, and decreased rate of CSF cryptococcal count within 2 weeks after admission. The areas under the curve (AUCs) in the internal, temporal, and external validation sets were 0.87 (95% CI 0.794-0.944), 0.92 (95% CI 0.795-1.000), and 0.86 (95% CI 0.744-0.975), respectively. An artificial intelligence (AI) model was trained to detect and count cryptococci, and the mean average precision (mAP) was 0.993. CONCLUSION: A ML model for predicting prognosis in HIV-negative CM patients was built and validated, and the model might provide a reference for personalized treatment of HIV-negative CM patients. The change in the CSF cryptococcal count in the early phase of HIV-negative CM treatment can reflect the prognosis of the disease. In addition, utilizing AI to detect and count CSF cryptococci in HIV-negative CM patients can eliminate the interference of human factors in detecting cryptococci in CSF samples and reduce the workload of the examiner.

Tree-based, two-stage risk factor analysis for postoperative sepsis based on Sepsis-3 criteria in elderly patients: A retrospective cohort study.

Background: Sepsis remains the leading cause of postoperative death in elderly patients and is defined as organ dysfunction with proven or suspected infection according to Sepsis-3 criteria. To better avoid potential non-linear associations between the risk factors, we firstly used a tree-based analytic methods to explore the putative risk factors of geriatric sepsis based on the criteria in the study. Methods: Data of 7,302 surgical patients aged ≥ 65 years at the Third Affiliated Hospital of Sun Yat-sen University from January 2015 to September 2020 were collected. An analytic method that combined tree-based analysis with the method of Mantel-Haenszel and logistic regression was adopted to assess the association between 17 putative risk factors and postoperative sepsis defined by the Sepsis-3 guideline by controlling 16 potential confounding factors. Results: Among the 16 potential covariates, six major confounders were statistically identified by the tree-based model, including cerebrovascular diseases, preoperative infusion of red blood cells, pneumonia, age ≥ 75, malignant tumor and diabetes. Our analysis indicated that emergency surgery increases the risk of postoperative sepsis in elderly patients by more than six times. The type of surgery is also a crucial risk factor for sepsis, particularly transplantation and neurosurgery. Other risk factors were duration of surgery > 120 min, administration of steroids, hypoalbuminemia, elevated creatinine, blood urea nitrogen, hematocrit, platelets, glucose, white blood cell count, abnormal neutrophil-to-lymphocyte ratio and elevated hsCRP-to-albumin ratio. Conclusions: Our study uses an effective method to explore some risk factors for postoperative sepsis in elderly by adjusting many potential confounders and it can provide information for intervention design.

G-CSF promotes the viability and angiogenesis of injured liver via direct effects on the liver cells.

BACKGROUND: Presently, liver transplantation is the only treatment strategy for liver failure (LF). Although granulocyte-colony stimulating factor (G-CSF) exhibits protective functions in LF, it is not clear whether it directly affects the liver cells. METHODS AND RESULTS: We established an injured liver cell model and observed that G-CSF treatment promoted cell viability and enhanced Ki67 and VEGF-A expression. Thereafter, human umbilical vein endothelial cells (HUVECs) were cultured in a conditioned medium collected from the G-CSF-treated injured liver cells. HUVECs' proliferation and tubule formation were promoted. Furthermore, in an injured liver mouse model, confirmed via haematoxylin-eosin staining, we evaluated serum alanine aminotransferase activity, Ki67 expression, and microvessel density (MVD). G-CSF treatment significantly relieved liver injury, upregulated Ki67 expression, and enhanced MVD in the injured mouse liver tissue. Additionally, AKT and ERK signal targets were explored, and it was demonstrated that the effects of G-CSF on injured liver cells were mediated through the AKT and ERK signalling pathways. CONCLUSIONS: G-CSF promotes injured liver viability and angiogenesis by directly affecting injured liver cells via the AKT and ERK signalling pathways. These findings improve our understanding of the role of G-CSF in recovery from LF.

Granulocyte Colony-Stimulating Factor Accelerates the Recovery of Hepatitis B Virus-Related Acute-on-Chronic Liver Failure by Promoting M2-Like Transition of Monocytes.

Background and Aim: Acute-on-chronic liver failure (ACLF) has a high mortality rate. The role of granulocyte colony-stimulating factor (G-CSF) in ACLF remains controversial. Monocytes/macrophages are core immune cells, which are involved in the initiation and progression of liver failure; however, the effect of G-CSF on monocytes/macrophages is unclear. The study aimed to verify the clinical efficacy of G-CSF and explore the effect of it on monocytes in hepatitis B virus (HBV)-related ACLF (HBV-ACLF) paitents. Methods: We performed a large randomized controlled clinical trial for the treatment of HBV-ACLF using G-CSF. A total of 111 patients with HBV-ACLF were prospectively randomized into the G-CSF group (5 µg/kg G-CSF every day for 6 days, then every other day until day 18) or the control group (standard therapy). All participants were followed up for at least 180 days. The relationship between monocyte count and mortality risk was analyzed. The effect of G-CSF on the phenotype and function of monocytes from patients with HBV-ACLF was evaluated using flow cytometry in vivo and in vitro experiments. Results: The survival probability of the G-CSF group at 180 days was higher than that of the control group (72.2% vs. 53.8%, P = 0.0142). In the G-CSF-treated group, the monocyte counts on days 0 and 7 were independently associated with an evaluated mortality risk in the fully adjusted model (Model 3) [at day 0: hazard ratio (HR) 95% confidence interval (CI): 15.48 (3.60, 66.66), P = 0.0002; at day 7: HR (95% CI): 1.10 (0.50, 2.43), P=0.8080]. Further analysis showed that after treatment with G-CSF in HBV-ACLF patients, the expression of M1-like markers (HLA-DR and CD86) in monocytes decreased (HLA-DR: P = 0.0148; CD86: P = 0.0764). The expression of MerTK (M2-like marker) increased (P = 0.0002). The secretion of TNF-α, IL-6, and IL-10 from monocytes decreased without lipopolysaccharide (LPS) stimulation (TNF-α: P < 0.0001; IL-6: P= 0.0025; IL-10: P = 0.0004) or with LPS stimulation (TNF-α: P = 0.0439; P = 0.0611; IL-10: P = 0.0099). Similar effects were observed in vitro experiments. Conclusion: G-CSF therapy confers a survival benefit to patients with HBV-ACLF. G-CSF can promote the anti-inflammatory/pro-restorative phenotype (M2-like) transition of monocytes, which may contribute to the recovery of ACLF.Clinical Trial Registration Number: ClinicalTrials.gov, identifier (NCT02331745).

Immunosuppressant and neuromyelitis optica spectrum disorder: optimal treatment duration and risk of discontinuation.

BACKGROUND AND PURPOSE: Preventing relapse by immunosuppressants (ISs) is critical for the prognosis of neuromyelitis optica spectrum disorder (NMOSD); however, the optimal duration of IS treatment is still under discussion. The objective was to explore the optimal duration of IS treatment and the risk of IS discontinuation for NMOSD. METHOD: This cohort study was conducted at a major neurological center that housed the largest NMOSD database in South China. Eligible participants were patients with NMOSD undergoing IS treatment. The main outcome measures were changes in relapse risk based on IS treatment duration, clinical outcomes and predictors of relapse following IS discontinuation. RESULTS: In total, 343 patients were included in this study. The duration of IS treatment was strongly associated with a decrease in relapse risk (hazard ratio [HR] 0.53, p < 0.001). Continuous IS treatment resulted in decreased relapse HRs within 5 years of receiving IS medication, with a mild rebound starting at 5 years. Rituximab reduced the risk of NMOSD relapse to approximately zero within 3 years. The rate of relapse after IS withdrawal was high (77.5%). As opposed to other ISs, a delayed relapse following rituximab withdrawal was observed in this study. Longitudinal extensive transverse myelitis (HR = 2.023, p = 0.006) was associated with a higher risk of relapse after IS discontinuation. CONCLUSIONS: Long-term IS medication for NMOSD is generally suitable. Patients with longitudinal extensive transverse myelitis had a higher risk of relapse after IS discontinuation. Future studies should explore individualized strategies of rituximab maintenance treatment.

Anticolon Cancer Targets and Molecular Mechanisms of Tao-He-Cheng-Qi Formula.

Background: Tao-He-Cheng-Qi Formula (THCQF) is a traditional Chinese medicine that has been proven to have antitumor effects. The aim of this study was to elucidate the molecular targets and mechanisms of THCQF against colon cancer and construct a prognostic model based on network pharmacology, bioinformatics analysis, and in vitro experiments. Methods: Potential THCQF compounds and targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine databases. Differentially expressed genes for colon cancer were screened in The Cancer Genome Atlas and Gene Expression Omnibus databases. The anticolon cancer mechanisms of THCQF were explored using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Molecular docking simulations and molecular dynamics analysis were used to evaluate the binding between target proteins and active compounds. Finally, the identified compounds were used to treat colon cancer cells from the HCT116 cell line, and expression of mRNA and protein after relevant posttreatment were tested using real-time polymerase chain reaction and western blotting. Results: A total of 27 anticolon cancer targets of THCQF were selected, among which four genes (CCNB1, CCNA2, IL1A, and MMP3) were shown to effectively predict patient outcomes in a prognostic colon cancer model. GO and KEGG enrichment analyses indicated that the activity against colon cancer of THCQF was associated with the interleukin (IL)-4 and IL-3 signaling pathways. Two compounds in THCQF, aloe emodin (AE) and quercetin (QR), were shown to efficiently bind to cyclin B1, the protein encoded by CCNB1. Finally, incubation of HCT116 cells with AE and QR significantly decreased CCNB1 mRNA expression and cyclin B1 levels. Conclusions: Taken together, the results indicate that AE and QR are the pivotal active compounds of THCQF, and CCNB1 is the main molecular target through which THCQF exerts its anticolon cancer effects. The study findings provide insight for studies investigating the anticancer effects of other traditional Chinese medicines.

Serum NfL associated with anti-NMDA receptor encephalitis.

INTRODUCTION: Neurofilament light chains (NfL) have been reported as potential markers for neuronal-axonal injury in neuroinflammatory diseases. In the current study, we describe serum NfL levels as a prognostic marker for anti-N-methyl-D-aspartate receptor encephalitis (NMDARE). METHODS: Serum levels of NfL of 64 patients with anti-NMDARE and 84 healthy controls were measured by Simoa. The anti-NMDAR Encephalitis One-Year Functional Status (NEOS) score, Modified Rankin Scale (mRS) scores, and clinical and cerebrospinal fluid parameters were evaluated in patients with anti-NMDARE. Meanwhile, we performed a receiver-operator characteristic analysis to assess the power of the serum NFL in predicting the 1-year functional status. RESULTS: Serum NfL levels were significantly elevated in patients with anti-NMDARE compared to healthy controls (p < 0.001, padjusted < 0.001), especially in patients with severe impairments (mRS > 3 vs ≤ 3, p = 0.035) or with limited response to treatment (vs. favorable outcome, p = 0.011). Serum NFL was positively associated with the initial admission mRS (r = 0.23, p = 0.072) and 1-year mRS (r = 0.29, p = 0.018). The AUC of serum NfL and NEOS score for 1-year poor functional status was 0.697 (95% CI 0.527-0.866, p = 0.011), 0.753 (95% CI 0.616-0.890, p = 0.001), respectively. Furthermore, AUC of the combination of serum NfL and NEOS was 0.815 (95% CI 0.680-0.950, p < 0.001). CONCLUSION: Our findings show that serum NfL levels evaluated in anti-NMDAR encephalitis may be a good predictor of the risk of 1-year poor functional status.

Fatigue Life Appraisal and Its Corrected Stress Intensity Factor for Repaired Off-CentrallyCracked Aluminum Plates.

This paper presents an experimental study on the fatigue life estimation of off-centrally cracked aluminum plates. Typical theoretical equations for off-central, central and edge cracks were reviewed and compared in terms of their sensitive parameters and applicability. A finite element model has been validated in its capacity in modelling the influences of eccentricity and crack size on the boundary correction coefficients. The Forman equation has been employed along with numerical results for the prediction of fatigue lives. Based on the test data, the fatigue life results of aluminum plates with and without patched laminate repair have been compared with codified fatigue classes. It is demonstrated that the repair at the crack tip close to the plate edge is effective in the fatigue life improvement for off-centrally crackedaluminum plates.

Comprehensive analysis of the long non-coding RNA-associated competitive endogenous RNA network reveals novel prognostic biomarkers in Wilms' tumor.

Wilms' tumor (WT) is one of the most common types of renal carcinoma in children. The aim of the present study was to construct a competitive endogenous RNA (ceRNA) regulation network and explore novel prognostic biomarkers for WT. The expression profiles were downloaded from The Cancer Genome Atlas database to identify differentially expressed RNAs (DERNAs). Based on the interactions between microRNAs (miRNAs) and mRNAs/long non-coding RNAs (lncRNAs), a ceRNA network was constructed. Functional enrichment analyses were subsequently conducted to explore the functions of the ceRNA-associated DEmRNAs. Survival analysis was performed to screen for prognosis-associated RNAs and the χ2 test was used to assess the associations between prognosis-associated RNA expression and histology classification/clinical staging. The present study identified 1,784 lncRNAs, 114 miRNAs and 3,337 mRNAs, which were abnormally expressed in WT compared with that in normal samples. By prediction, pairing and network analysis, a ceRNA network consisting of 38 DElncRNAs, 18 DEmiRNAs and 99 DEmRNAs was established. These DEmRNAs were significantly enriched in pathways associated with the occurrence and development of WT. By combining the expression data with survival analysis, seven prognosis-associated RNAs were identified (P<0.05). Of these seven RNAs, two (zinc finger and BTB domain containing 4; and deleted in lymphocytic leukemia 2) were significantly associated with clinical staging and histology classification. Lastly, the expression levels of the seven RNAs were verified in the Gene Expression Omnibus database. The present study revealed that 7 RNAs might be considered as novel prognostic biomarkers and potential treatment targets for therapy in WT. In addition, the ceRNA regulation network could provide novel strategies for further studies on lncRNAs and miRNAs in WT.

Association between Eating Away from Home and Hyperuricemia: A Population-Based Nationwide Cross-Sectional Study in China.

Hyperuricemia (HU) is a risk factor for different kinds of chronic noncommunicable diseases, and eating away from home (EAFH) may play an important role in their development, which has been ignored greatly so far. This study aimed to investigate the association between EAFH and HU in different models. A cross-sectional study involving 8,322 participants of the China Health and Nutrition Survey (CHNS) was conducted. Logistic regression models were used to analyze the data. We found that participants who consumed more away-from-home food had a higher risk for HU, and the adjusted odds ratio (aOR) and 95% confidence interval (CI) (for each increment in grades of EAFH) were 1.11 (1.02, 1.20) in a multiadjusted model (adjusted for age, gender, province, net individual income, body mass index, smoking, leisure-time physical activities, energy intake, and sleep duration). As for stratified analyses, the aOR (95% CI) of EAFH was 1.12 (1.01, 1.24) for men and 1.06 (0.92, 1.21) for women. Similar results can be found in the middle-aged and obese population, with aOR (95% CI) of EAFH as 1.17 (1.05, 1.30) and 1.15 (1.03, 1.29), respectively. In conclusion, EAFH is positively associated with the prevalence of HU.

Clinical significance of tumor necrosis factor receptor 2 in middle and lower thoracic esophageal squamous cell carcinoma.

High expression and role of tumor necrosis factor receptor 2 (TNFR2) in cancer progression and prognosis has been reported in several types of tumors. However, its role in esophageal carcinoma (EC) remains unknown. In the present study, TNFR2 expression in middle and lower thoracic esophageal squamous cell carcinoma (ESCC) was detected using immunohistochemistry (IHC). Chi-square test revealed that TNFR2 was positively correlated with invasion depth, advanced clinical stage and low differentiation degree. Furthermore, survival analysis revealed that TNFR2 was positively correlated with poor overall survival (OS). Moreover, univariate COX regression analysis revealed that clinical stage, lymph node involvement, and invasion depth can affect the OS of ESCC patients, while multivariate COX regression analysis revealed that lymph node involvement and invasion depth can affect the OS of ESCC patients. In middle thoracic ESCC patients, TNFR2 was positively correlated with invasion depth, advanced clinical stage and poor OS. Furthermore, univariate and multivariate COX regression analysis both revealed that clinical stage, lymph node involvement, and invasion depth can affect OS. In lower thoracic ESCC patients, TNFR2 was positively correlated with low differentiation degree. Furthermore, the positive correlation of TNFR2 with poor OS did not reach statistical significance. In addition, univariate COX regression analysis revealed that only lymph node involvement could affect OS. All the results suggest that TNFR2 can play an important role in the progression and poor prognosis of ESCC patients. Moreover, the role of TNFR2 in the prognosis of middle thoracic ESCC patients was earlier and stronger than in lower thoracic ESCC patients.

High expression of tumor necrosis factor receptor 2 in tissue is associated with progression and prognosis of esophageal squamous cell carcinoma.

In recent years, the role of TNFR2 has attracted much attention for its promotion role in several types of tumors. However, in ESCC, the clinical relevance of TNFR2 is still unknown. In this study, we detected TNFR2 expression in ESCC tissues using immunohistochemistry. The χ2 test showed that TNFR2 was positively correlated with invasion depth, advanced clinical stage, and low differentiation; survival analysis showed that TNFR2 was positively correlated with poor OS; univariate Cox regression analysis showed that clinical stage, lymph node involvement, and invasion depth were all correlated with OS; and multivariate Cox regression analysis showed that lymph node involvement and invasion depth were independent prognostic factors. In male cases, TNFR2 was positively correlated with invasion depth, advanced clinical stage, low differentiation, and poor OS; univariate Cox regression analysis showed that clinical stage, lymph node involvement, and invasion depth were all associated with OS; and multivariate Cox regression analysis showed that lymph node involvement was an independent prognostic factor. In female cases, TNFR2 was positively correlated with invasion depth, advanced clinical stage, and poor OS; univariate Cox regression analysis showed that only lymph node involvement was associated with OS. All the results confirmed that TNFR2 in ESCC tissues was positively correlated with progression and poor prognosis of ESCC patients. Lymph node involvement and invasion depth can be treated as independent prognostic factors.

Cellular behaviours of bone marrow-derived mesenchymal stem cells towards pristine graphene oxide nanosheets.

OBJECTIVES: Graphene oxide (GO), the derivative of graphene with unique properties, has attracted much attention for applications in dental implants. The aim of this study was, by two biomimetic cell culture methods, to investigate the quantitative relationship between the concentration of pristine GO nanosheets and their cellular behaviours towards bone marrow-derived mesenchymal stem cells (BMSCs). MATERIALS AND METHODS: The cells were firstly characterized according to their morphology, self-renewal capabilities and multipotency. Subsequently, adhesion density, proliferation, alkaline phosphatase activity and mineralization of BMSCs treated with various concentrations of GO were analysed. In addition, osteogenic-related proteins were measured for further verification of the GO-induced osteogenic differentiation. RESULTS: Pristine GO nanosheets inhibited the proliferation of BMSCs at a high concentration of 10 µg/mL during the first 3 days with two seeding methods and facilitated proliferation of BMSCs at a low concentration of 0.1 µg/mL after 5 days with a sequential-seeding method compared to a co-seeding method. Analogously, osteogenic differentiation was promoted when BMSCs were treated with 0.1 µg/mL of GO. Both the proliferation and differentiation showed concentration-dependent behaviour. Interestingly, Wnt/ß-catenin signalling pathway appeared to be involved in osteogenic differentiation induced by pristine GO nanosheets. CONCLUSIONS: Pristine GO nanosheets at a concentration of 0.1 µg/mL provide benefits to promote BMSCs proliferation and osteogenesis under a sequential-seeding method, contributing to the use of GO for dental implantation.

Tumor necrosis factor receptor 2 promotes growth of colorectal cancer via the PI3K/AKT signaling pathway.

Tumor necrosis factor receptor 2 (TNFR2) is the receptor for tumor necrosis factor α (TNF-α). TNFR2 differs from tumor necrosis factor 1 (TNFR1) in various ways and is mainly expressed in hematopoietic and endothelial cells. However, studies about its functions in tumors are limited. The contributions of TNFR2 in colorectal cancer (CRC) remain unknown. In the present study, it was found that TNFR2 was positively associated with Ki67 expression in CRC tissues using immunohistochemistry (IHC), and western blot analysis found that Ki67 was upregulated by overexpressing TNFR2 in SW1116 cells and inhibited by silencing TNFR2 in HT29 cells. Methyl thiazolyl tetrazolium assay found that growth of SW1116 cells overexpressing TNFR2 was significantly increased compared with the control group and that the growth of HT29 cells subsequent to silencing TNFR2 was significantly decreased compared with the control group. Clone formation assay found that more clones were formed in SW1116 cells overexpressing TNFR2 than the control group, and less clones formed in HT29 cells subsequent to silencing TNFR2 than the control group. In addition, western blot analysis found that phosphorylation of protein kinase B (AKT) was activated subsequent to overexpressing TNFR2 in SW1116 cells, and inhibited following silencing of TNFR2 in HT29 cells. Additionally, treatment using LY294002 significantly abrogated the promotion of Ki67 expression, growth and clone formation abilities induced by TNFR2 overexpression in SW1116 cells. All the results suggest that TNFR2 can significantly promote CRC growth via the phosphoinositide 3-kinase/AKT signaling pathway; this provides evidential support for taking TNFR2 as a new target for CRC treatment.

Downregulation of miR-221/222 by a microRNA sponge promotes apoptosis in oral squamous cell carcinoma cells through upregulation of PTEN.

MicroRNA-221 and microRNA-222 (miR-221/222) have been identified as oncogenes and confirmed to be overexpressed in various types of cancer. However, the regulation mechanism of miR-221/222 in oral squamous cell carcinoma (OSCC) remains to be fully elucidated. Previously, an miR-221/222 sponge was successfully constructed and its effect on the downregulation of miR-221/222 expression was investigated. In the present study, the dual luciferase reporter assay revealed a phosphatase and tensin homolog (PTEN) deletion on chromosome 10 to be a target gene of miR-221/222. It was also demonstrated that miR-221/222 suppression by transfection with an miR-221/222 sponge in vitro resulted in upregulation of PTEN. Notably, the proliferation and invasiveness of the miR-221/222 sponge-transfected cells was significantly inhibited, while apoptosis was promoted, when determined by Cell Counting Kit-8, Transwell assays and flow cytometry. The results of the present study prove that miR-221/222 may downregulate the expression of PTEN in OSCC cells and function as oncogenes, providing a novel insight into the underlying mechanism of OSCC tumorigenesis. The present study suggests that upregulating the expression of PTEN by downregulation of miR-221/222 may be a potential treatment for OSCC.

Resveratrol inhibits inflammation and ameliorates insulin resistant endothelial dysfunction via regulation of AMP-activated protein kinase and sirtuin 1 activities.

BACKGROUND: Resveratrol is a phytoalexin with beneficial effects on human health. The aim of the present study was to investigate the effects of resveratrol on endothelial dysfunction involved in insulin signaling and inflammation. METHODS: Endothelial cells were stimulated with palmitate (PA) to induce insulin resistance characterized by a loss of insulin-mediated nitric oxide (NO) production. Diabetes was induced in rats by fructose feeding. The effects of resveratrol and the mechanisms involved were investigated using an aortic relaxation assay and Western blot analysis. RESULTS: In endothelial cells, 0.1-10 µmol/L resveratrol suppressed IκB kinase ß (IKKß)/nuclear factor-κB phosphorylation, as well as tumor necrosis factor-α and interleukin-6 production, and restored the insulin receptor substrate-1 (Irs-1)/Akt/endothelial NO synthase signaling pathway. Furthermore, resveratrol effectively inhibited the mitogenic actions of insulin by decreasing the secretion of endothelin-1 and plasminogen activator inhibitor-1. It also positively regulated AMP-activated kinase (AMPK) and sirtuin 1 (SIRT1) activation, which contributed to the inhibition of inflammation implicated in endothelial insulin resistance. Stimulation with PA and long term-fructose feeding impaired insulin-mediated vessel dilation in rat aorta, whereas pretreatment of aortic rings with resveratrol (0.1-10 µmol/L) or treatment of rats with 5 or 20 mg/kg resveratrol counteracted these changes. CONCLUSION: The results indicate that resveratrol inhibits inflammation and facilitates insulin phosphatidylinositol 3-kinase signaling by beneficial modulation of IRS-1 function partly via regulation of AMPK and SIRT1 activity in the endothelium.

MiR-222 targeted PUMA to improve sensitization of UM1 cells to cisplatin.

microRNAs have been shown to play critical roles in regulating the chemosensitivity of cancer cells. As a member of the oncogenic miRNAs (oncomiRs), miR-222 has been reported to drive the oncogenesis of many types of malignancies. However, little is known concerning the specific role of miR-222 in human oral squamous cell carcinoma (OSCC). The present study explored the role and mechanism of miR-222 in increasing the expression of p53 up-regulated modulator of apoptosis (PUMA) and enhancing the sensitivity of OSCC to cisplatin (CDDP). Results showed that antisense (As)-miR-222 inhibits the expression of miR-222. In contrast, PUMA was dramaticallyup-regulated. IC50 values were significantly decreased in cells treated with As-miR-222 combined with CDDP, to a greater extent than in cells treated with CDDP alone. Furthermore, As-miR-222 enhanced apoptosis and inhibited the invasiveness of UM1 cells. Analysis of the above data suggested that, in UM1 cells, there might be a regulatory loop between miR-222 and PUMA, and that miR-222 inhibition increased the chemosensitivity to CDDP. These findings demonstrated that down-regulation of miR-222 could enhance the chemosensitivity of human OSCC cells to CDDP, and that the combination of As-miR-222 and CDDP could be an effective therapeutic strategy by boosting the expression of PUMA for controlling the growth of OSCC.

miR-222 regulates the cell biological behavior of oral squamous cell carcinoma by targeting PUMA.

Previous reports have shown that low expression of p53 upregulated modulator of apoptosis (PUMA) and abnormal expression patterns of a number of miRNAs may be associated with poor prognosis in various types of human malignancies. As a member of the oncomiRs, miR-222 has been found to be upregulated in oral squamous cell carcinoma (OSCC). We hypothesized that there was an important relationship between miR-222 and PUMA in OSCC based on the prediction of the target genes of miR-222. In the present study, Pre-miR-222, As-miR-222 and the empty vector, were used to treat OSCC cells, respectively. Using the non-transfected cells as blank control, the expression levels of miR-222 and the PUMA gene were evaluated by RT-PCR and western blotting. Cell proliferation and migration abilities were analyzed by MTT and Transwell assays. Cell cycle distribution and apoptosis were assessed by flow cytometry. Our results demonstrated that, when compared with the blank control group, OSCC cells in the Pre-miR-222 transfection group showed increased expression of miR-222 and decreased expression of PUMA, enhanced proliferation and invasion abilities, and decreased apoptosis. In contrast, the above indices in the As-miR-222 transfection group confirmed the opposite results when compared with those in the Pre-miR-222 transfection group. In addition, no significant differences between the empty vector transfection group and the control group were noted. Our results suggest that miR-222 targets the expression of PUMA in OSCC cells and affects cell growth, invasive and apoptotic abilities. Thus, PUMA may be a possible new target for the treatment of OSCC.