MicroRNA-513b-5p inhibits epithelial mesenchymal transition of colon cancer stem cells through IL-6/STAT3 signaling pathway.

OBJECTIVE: To reveal the mechanisms by which miR-513b-5p inhibits metastasis of colon cancer stem cells (CCSCs) through IL-6/STAT3 in HCT116 cells. METHODS: Sphere formation media and magnetic cell sorting were used to enrich and screen CCSCs. We used a colony formation assay, cell proliferation and viability assays, and a nude mouse transplantation tumor assay to identify CCSCs. ELISA was performed to identify IL-6 in the cell culture medium, and the growth, viability, wound healing, and transwell migration of distinct cell groups were compared to differentiate them. Dual-luciferase reporter assay, RT-PCR, and/or Western Blot analysis were conducted to determine the correlation between them. RESULTS: CD133+CD44+ HCT116 cells were shown to have higher cloning efficiency, greater proliferation ability and viability, and stronger tumorigenicity. A dual-luciferase reporter assay revealed that miR-513b-5p negatively affected STAT3 expression. RT-PCR and/or Western Blot analysis suggested that miR-513b-5p negatively affected STAT3 and Vimentin, while positively affecting E-cadherin expression. The STAT3 overexpression vector + miR-513b-5p inhibitor cell group had the highest efficiency, greatest proliferation ability and viability, and the highest IL-6 level in the experiments. CONCLUSIONS: Mir-513b-5p inhibited the epithelial-mesenchymal transition (EMT) of CCSCs through IL-6/STAT3. This potential mechanism may provide a new therapeutic target for colon cancer.

Relationship Between Ultra-Processed and Minimally Processed Food Intake and Cardiovascular Health Among US Women of Reproductive Age.

Objectives: Ultra-processed food (UPF) intake is associated with worse cardiovascular health (CVH), but associations between unprocessed/minimally processed foods (MPFs) and CVH are limited, especially among women of reproductive age (WRA). Materials and Methods: For 5,773 WRA (20-44 years) in National Health and Nutrition Examination Survey (NHANES) 2007-2018, we identified UPFs and MPFs using the Nova classification and based on 24-hour dietary recalls. We calculated usual percentages of calories from UPFs and MPFs using the National Cancer Institute's usual intake method. Seven CVH metrics were scored, and CVH levels were grouped by tertile. We used multivariable linear and multinomial logistic regression to assess associations between UPFs and MPFs and CVH. Results: The average usual percentage of calories from UPFs and MPFs was 57.2% and 29.3%, respectively. There was a graded, positive association between higher UPF intake and higher odds of poor CVH: adjusted odds ratios (aORs) for the lowest versus highest CVH were 1.74 (95% confidence interval: 1.51-2.01), 2.67 (2.07-3.44) and 4.66 (3.13-6.97), respectively, comparing quartile 2 (Q2)-Q4 to the lowest quartile (Q1) of UPF intake. Higher MPF intake was associated with lower odds of poor CVH: aORs for the lowest CVH were 0.61 (0.54-0.69), 0.39 (0.31-0.50), and 0.21 (0.14-0.31). Patterns of association remained consistent across subgroups and in sensitivity analyses. Conclusions: Higher UPF intake was associated with worse CVH, while higher MPF intake was associated with better CVH among WRA in the United States. Our analyses highlight an opportunity for WRA to improve nutrition and their CVH.

Multivariate analysis of prognostic factors in patients with lung cancer.

Objective: Lung cancer is the leading cause of cancer-related mortality in China. The purpose of this study was to determine the effect of non-therapeutic and therapeutic factors of patients with lung cancer on survival rate. Methods: In this retrospective study, a total of 458 patients diagnosed as lung cancer at the Department of Thoracic Surgery, the Fourth Affiliated Hospital of Hebei Medical University from September 2008 to October 2013 were enrolled. The COX proportional hazards model was used to analyze the possible factors affecting the survival of patients. Model variables included age, sex, family history, smoking, tumor location, pathological type, stage, chemotherapy, radiotherapy, operation, and targeted therapy. Results: The median survival time (MST) was 32.0 months (95% CI: 29.0-34.0 months), while the 1-, 3-, and 5-year survival rates were 70.74%, 36.90%, and 30.13%, respectively. The univariate analysis showed that stage, chemotherapy, radiotherapy, and operation significantly affected the median survival time of patients. Multivariate cox regression analysis suggested that sex (female vs male, 2.096, 95% CI: 1.606-2.736), stage (stage I vs IV, 0.111, 95% CI: 0.039-0.314; stage II vs IV, 0.218, 95%CI: 0.089-0.535), chemotherapy (no vs yes, 0.469, 95% CI: 0.297-0.742), and operation (no vs yes, 2.667, 95% CI: 1.174-6.055) were independently associated with the survival of patients with lung cancer. Conclusion: Our study showed that male, early stage, operation were protective factors for the survival of patients, while female, advanced stage, chemotherapy were risk factors for the survival of patients. Larger studies are required to address the usefulness of these prognostic factors in defining the management of patients with lung cancer.

Chronic atrophic gastritis in different ages in South China: a 10-year retrospective analysis.

OBJECTIVES: To explore the prevalence, characteristics, age distribution and etiology changes of chronic atrophic gastritis (CAG) in South China. METHODS: This study included all patients who underwent endoscopy examinations from 2011 to 2020 in our hospital. Patients were divided into groups 1 (2011-2015) and 2 (2016-2020). The prevalence, characteristics, age distribution and etiology changes of CAG were compared between groups. RESULTS: Overall CAG prevalence was 20.92% (24,084/115,110) from 2011 to 2020; prevalence significantly differed between groups (18.78%, 8468/45,087, in group 1 and 22.30%, 15,616/70,023, in group 2). Patients with CAG had significantly younger age (under 45) and more corpus atrophy and more autoimmune atrophic gastritis (AAG) in group 2 than in group 1. AAG prevalence in group 2 was 30.11% (4702/15,616) significantly higher than 13.57% (1149/8468) in group 1. 82 patients with AAG later exhibited gastric cancer without obvious clinical features over the decade. CONCLUSIONS: CAG is increasing and seems starting earlier among people during the study period. We need to focus on diagnosis and treatment of corpus related atrophy and AAG, especially for the young. Laboratory examination, endoscopic biopsy and surveillance are important for CAG.

Tubiechong patching promotes tibia fracture healing in rats by regulating angiogenesis through the VEGF/ERK1/2 signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The external use of traditional Chinese medicine (TCM) to treat fractures has a long history of clinical application and theoretical basis, and is also one of the characteristic treatment methods of TCM with significant efficacy and many advantages. Among the commonly used external Chinese medicines, Tubiechong is noteworthy. AIM OF THE STUDY: To elucidate whether local patching of Tubiechong can promote fracture healing and explore its mechanism of action. MATERIALS AND METHODS: A rat tibia fracture model was constructed by the modified Einhorn modeling method. X-ray films were taken to evaluate the progress of fracture healing. Serum bone alkaline phosphatase (BALP), osteocalcin (BGP) and the C-terminal content of collagen type I (CTX-I) were analyzed by ELISA. CD31 immunohistochemistry was used to evaluate angiogenesis in the tibia segment. The effects of Tubiechong decoction (TD) on HUVEC proliferation, migration and invasion were detected by MTT assay, wound healing assay and Transwell migration assay, respectively. RNA-seq was performed to identify differentially expressed genes (DEGs). Enrichment of functions and signaling pathway analysis were performed based on the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Quantitative real time polymerase chain reaction (qRT-PCR) was used to study gene expression levels. Western blotting (WB) was used to detect the expression of relevant regulatory proteins. RESULTS: The healing time of rat tibia fractures in the three TD dose groups was shortened. The serum levels of BALP, BGP and CTX- I in the TD-treated group were higher than those in the NC group. The X-ray results showed that on the 7th day after surgery, the fracture healing degree of the high-dose TD group was significantly better than that of the NC group, and the fracture healing degrees of each TD treatment group were significantly higher than those of the NC group on the 14th, 17th, and 21st days after the operation. The CD31 immunohistochemistry results showed that the number of blood vessels and the vascular area in the TD treatment group were higher than those in the NC group. In vitro, TD promoted the proliferation, wound healing and migration of HUVECs. GO analysis of transcriptome sequencing results showed that TD significantly altered the expression of genes related to cell growth, metabolism, and motility. According to KEGG annotations, VEGFA was upregulated. Eight DEGs were enriched in the VEGFA-VEGFR2 signaling pathway, of which six were upregulated. KEGG signaling pathway analysis showed that the most abundant DEGs were in mitogen-activated protein kinase (MAPK) signaling pathway. qRT-PCR showed that VEGFA gene expression in HUVECs was 7.8 times that of the control group after 1 mg/mL TD treatment for 24 h, and WB experiments showed that its protein expression was 3 times that of the control group. WB results showed that the phosphorylated ERK gene was highly expressed, while the expression levels of phosphorylated P38 and phosphorylated JNK protein remained unchanged. CONCLUSION: Tubechong patching therapy promotes tibia fracture healing in rats by regulating angiogenesis through the VEGF/ERK1/2 signaling pathway.

Comparison of the efficacy of small-incision clamp-assisted reduction and open reduction for the treatment of femoral shaft fractures with an anterograde intramedullary nail: a retrospective study.

BACKGROUND: To compare the efficacy of small-incision clamp-assisted reduction with open reduction for the treatment of femoral shaft fractures by anterograde intramedullary nailing. METHODS: The data of 63 patients with femoral shaft fractures, treated between January 2016 and June 2021, were retrospectively analyzed. All patients received anterograde intramedullary nail fixation, and the OA/OTA classification of fractures was 32-C. The average follow-up period was 13 months (range: 11-14 months). According to the method of fracture reduction, patients were divided into a small-incision clamp-reduction group (referred to as the clamp-reduction group) and an open-reduction group. The reduction time, operative time, the number of fluoroscopy, intraoperative blood loss, postoperative VAS score, postoperative time to discharge, and the rates of intraoperative and postoperative complications were compared between the two groups. RESULTS: There were statistically significant differences in reduction time, operative time, the number of fluoroscopy, intraoperative blood loss, postoperative VAS score, postoperative time to discharge (t = 6.718, - 11.679, 18.963, - 11.609, - 22.432, - 7.187; P < 0.05). In the clamp-reduction group, there was no intraoperative blood transfusion. However, there were one case of wound infection and one case of deep vein thrombosis after operation. In the open-reduction group, ten patients received intraoperative blood transfusion, one patient developed hemorrhagic shock, two patients developed wound infection, and two patients developed bone nonunion during follow-up. CONCLUSIONS: Both groups had good functional recovery after operation. However, compared with open reduction, clamp reduction is a safer reduction method with shorter operation time, less intraoperative blood loss, less postoperative pain, shorter hospital stay and fewer postoperative complications.

Relationship Between Ultraprocessed Food Intake and Cardiovascular Health Among U.S. Adolescents: Results From the National Health and Nutrition Examination Survey 2007-2018.

PURPOSE: Studies of the association between ultraprocessed foods (UPF) and cardiovascular disease risk factors have been mainly focused on the adult population. This study examined the association between usual percentage of calories (%kcal) from UPF and the American Heart Association's seven cardiovascular health (CVH) metrics among U.S. adolescents aged 12-19 years. METHODS: We used data from the National Health and Nutrition Examination Survey 2007-2018 (n = 5,565). The NOVA food system was used to classify UPF according to the extent and purpose of food processing. Each CVH metric was given a score of 0, 1, or 2 (poor, intermediate, or ideal health, respectively). Scores of six metrics were summed (excluding diet) to categorize CVH as low (0-7), moderate (8-10), or high (11-12). The National Cancer Institute's methods were used to estimate usual %kcal from UPF. Multivariable linear regression and multinomial logistic regression were used to evaluate the association between UPF and CVH. RESULTS: Among youth, 12.1% had low CVH, 56.3% moderate, and 31.6% high. The mean usual %kcal from UPF was 65.7%. Every 5% increase in calories from UPF was associated with .13 points lower CVH scores (p < .001). Comparing Q2, Q3, and Q4 to Q1 of UPF intake, the adjusted odds ratios for low versus high CVH were 1.43 (95% confidence interval 1.16-1.76), 1.86 (1.29-2.66), and 2.59 (1.49-4.55), respectively. The pattern of association was largely consistent across subgroups. CONCLUSIONS: U.S. adolescents consume about two thirds of daily calorie from UPF. There was a graded inverse association between %kcal from UPF and CVH score.

Association between ultraprocessed food intake and cardiovascular health in US adults: a cross-sectional analysis of the NHANES 2011-2016.

BACKGROUND: Higher intake of ultraprocessed foods (UPFs) might be associated with increased risk of cardiovascular disease. OBJECTIVES: Our objective was to examine the association between usual percentage of calories (%kcal) from UPFs and the American Heart Association's "Life's Simple 7" cardiovascular health (CVH) metrics in US adults. METHODS: We analyzed data from 11,246 adults aged ≥20 y from the NHANES 2011-2016 (a cross-sectional, nationally representative survey). UPF designation was assigned on the basis of the NOVA classification system, according to the extent and purpose of food processing. Each CVH metric was given a score of 0, 1, or 2 representing poor, intermediate, or ideal health, respectively. Scores of the 6 metrics (excluding diet) were summed, and CVH was categorized as inadequate (0-4), average (5-8), or optimum (9-12). We used the National Cancer Institute's methods to estimate the usual %kcal from UPFs, and multivariable linear and multinomial logistic regression to assess the association between UPFs and CVH, adjusted for age, sex, race and Hispanic origin, education, and poverty. RESULTS: The weighted prevalence of inadequate, average, and optimum CVH was 8.0%, 51.7%, and 40.3%, respectively. The mean usual %kcal from UPFs was 55.4%, and midpoint of quartiles of intake ranged from 40.4% (quartile 1) to 70.5% (quartile 4). Every 5% increase in calories from UPFs was associated with 0.14 points lower CVH score (P < 0.001). The adjusted ORs for inadequate CVH were 1.40 (95% CI: 1.23, 1.60), 1.82 (1.45, 2.29), and 2.57 (1.79, 3.70), respectively, comparing quartiles 2, 3, and 4 with quartile 1 of UPF intake. The pattern of association was largely consistent across subgroups. CONCLUSIONS: Usual %kcal from UPFs represented more than half of total calorie intake in US adults. A graded inverse association between %kcal from UPFs and CVH was observed.

Correlation of gene polymorphisms of vascular endothelial growth factor with grade and prognosis of lung cancer.

BACKGROUND: Vascular endothelial growth factor (VEGF) gene is highly polymorphic, and single nucleotide polymorphisms (SNP) of VEGF gene are associate with cancer prognosis. This study aimed to analyze the correlation of VEGF gene polymorphisms with grade and prognosis of lung cancer. METHODS: A total of 458 Chinese patients with primary lung cancer were enrolled from September 2008 to October 2013. The genotypes of -2578C > A, -1154G > A, - 460 T > C, and + 405G > C were analyzed in white blood cells from patients using polymerase chain reaction based restriction fragment length polymorphism. RESULTS: Our data showed that -1154G > A polymorphism was significantly associated with tumor stages, but all four tested VEGF gene polymorphisms had no significant effect on survival. CONCLUSIONS: VEGF polymorphisms may relate to stage of lung cancer in Chinese population.

Prevalence and Cardiovascular Health Impact of Family History of Premature Heart Disease in the United States: Analysis of the National Health and Nutrition Examination Survey, 2007-2014.

Background Because family history is a known risk factor for heart disease, it is important to characterize its public health impact in terms of population prevalence of family history of heart disease, the burden of heart disease attributable to family history, and whether family history interacts with modifiable risk factors for heart disease. Methods and Results We used population data from NHANES (the National Health and Nutrition Examination Survey [2007-2014]) to measure the association of self-reported family history of premature heart disease ( FHPHD ) with cardiovascular disease (n=19 253) and to examine the association between cardiovascular health metrics and FHPHD (n=16 248). Using logistic regression and multivariable adjustment, family history odds ratios were 5.91 (95% CI , 3.34-10.44) for ages 20 to 39, 3.02 (95% CI, 2.41-3.79) for ages 40 to 59, and 1.87 (95% CI , 1.54-2.28) for age ≥60 for cardiovascular disease. The prevalence of cardiovascular disease for the population with a FHPHD (15.72%; 95% CI , 13.81-17.64) was more than double the prevalence of cardiovascular disease for those without a family history (6.25%; 95% CI , 5.82-6.69). Compared with participants with optimum cardiovascular health, the prevalence ratio for FHPHD was 1.98 (95% CI , 1.40-2.79) for those with inadequate cardiovascular health. Conclusions Millions of people who are at high risk of having cardiovascular disease could be identified using FHPHD . FHPHD can become an important component of public health campaigns that address modifiable risk factors that plan to reduce the overall risk of heart disease.

Cardiovascular Health Among Non-Hispanic Asian Americans: NHANES, 2011-2016.

Background Asian Americans are the fastest growing population in the United States, but little is known about their cardiovascular health (CVH). The objective of this study was to assess CVH among non-Hispanic Asian Americans (NHAAs) and to compare these estimates to those of non-Hispanic white (NHW) participants. Methods and Results Merging NHANES (National Health and Nutrition Examination Survey) data from 2011 to 2016, we examined 7 metrics (smoking, weight, physical activity, diet, blood cholesterol, blood glucose, and blood pressure) to assess CVH among 5278 NHW and 1486 NHAA participants aged ≥20 years. We assessed (1) the percentage meeting 6 to 7 metrics (ideal CVH), (2) the percentage meeting only 0 to 2 metrics (poor CVH), and (3) the overall mean CVH score. We compared these estimates between NHAAs and NHWs and among foreign-born NHAAs by birthplace and number of years living in the United States. The adjusted prevalence of ideal CVH was 8.7% among NHAAs and 5.9% among NHWs ( P<0.001). NHAAs were significantly more likely to have ideal CVH (adjusted prevalence ratio: 1.42; 95% CI, 1.29-1.55) compared with NHWs. Among NHAAs, there was no significant difference in ideal CVH between US- and foreign-born participants, nor by number of years living in the United States. With lower body mass index thresholds (<23, normal weight) for NHAAs, there were no statistically significant differences in the adjusted prevalence of ideal CVH (6.5% versus 5.9%, P=0.216) between NHAAs and NHWs. Conclusions NHAAs had a higher prevalence of overall ideal CVH compared with NHWs. However, when using a lower body mass index threshold for NHAAs, there was no difference in ideal CVH between the groups.

MicroRNA-485-5p suppresses the proliferation, migration and invasion of small cell lung cancer cells by targeting flotillin-2.

This study is aimed to elucidate the mechanisms underlying the role of miR-485-5p in small cell lung cancer (SCLC). The expression of miR-485-5p were quantified with real time quantitative PCR and it was found that the level of miR-485-5p was lower in SCLC tissues than normal tissues. In cultured SCLC cell lines, overexpression of miR-485-5p reduced cell proliferation, migration, and invasion in vitro, whereas knockdown of miR-485-5p performed contrary. FLOT2 expression was obviously upregulated and negatively correlated with miR-485-5p expression level in SCLC tissues. Overexpression of miR-485-5p significantly inhibited the protein expression of flotillin-2 (FLOT2) in cultured SCLC cells. Luciferase reporter assay confirmed that FLOT2 was a direct target of miR-485-5p in SCLC cells. It is concluded that miR-485-5p, as a tumor suppressor, inhibits the growth and metastasis in SCLC by targeting FLOT2. Upregulation of miR-485-5p expression may be an attractive strategy for SCLC therapy.

Association between cardiovascular health metrics and depression among U.S. adults: National Health and Nutrition Examination Survey, 2007-2014.

PURPOSE: The American Heart Association has identified seven modifiable cardiovascular health (CVH) metrics, including four health behaviors (body mass index, smoking, physical activity, and dietary intake) and three health factors (total cholesterol, blood pressure, and fasting glucose). We sought to examine the association between CVH metrics and depression. METHODS: We analyzed data on 14,561 adults aged 20 years or older from the National Health and Nutrition Examination Survey 2007-2014. Depressive symptoms were assessed using the Patient Health Questionnaire; a score of 0-4, 5-9, and 10 or higher represented no or minimal, mild, moderate or severe depressive symptoms, respectively. CVH was categorized as inadequate, average, or optimum. We used multinomial logistic regression to assess the association between CVH and depression, adjusted for age, gender, race or ethnicity, education, and alcohol use. RESULTS: Prevalence of inadequate, average, and optimum CVH were 6.1%, 59.7%, and 34.2%; 14.9% and 7.8% of adults had mild and moderate/severe depression, respectively. Compared with participants with optimum CVH, prevalence ratios for moderate or severe depression were 4.39 (95% confidence interval, 3.32-5.80) and 2.64 (2.15-3.24) for those with inadequate and average CVH, respectively. The corresponding prevalence ratios for mild depression were 2.11 (1.77-2.52) and 1.36 (1.19-1.55). The association appeared to be stronger for CVH behaviors. CONCLUSIONS: There was a graded association between CVH metrics, particularly for health behaviors, and mild and moderate/severe depression among U.S. adults.

Income Disparities and Cardiovascular Risk Factors Among Adolescents.

BACKGROUND AND OBJECTIVES: Socioeconomic disparities in cardiovascular health among adults have been documented, but disparities during adolescence are less understood. In this study, we examined secular trends in cardiovascular risk factors and disparities among US adolescents. METHODS: We analyzed NHANES data from 1999 to 2014, including 11 557 (4854 fasting) participants aged 12 to 19 years. To examine trends in cardiovascular risk factors, adolescents were stratified into 3 groups on the basis of family poverty-income ratio: low income (poverty-income ratio, <1.3), middle income (≥1.3 and <3.5), and high income (≥3.5). RESULTS: From 1999 to 2014, the prevalence of obesity increased (16.3%-20.9%, P = .001) but only among low- and middle-income adolescents, with significant disparities in prevalence by income (21.6% vs 14.6% among low- versus high-income adolescents, respectively, in 2011-2014). In addition, there were significant and persistent disparities in the prevalence of smoking (20.7% vs 7.3% among low- versus high-income adolescents, respectively, in 2011-2014), low-quality diet (68.9% vs 55.4%), and physical inactivity (25.6% vs 17.0%). No significant disparities were observed in the prevalence of prediabetes and diabetes, hypertension, or hypercholesterolemia, although the prevalence of prediabetes and diabetes nearly doubled (11.9%-23.1%, P < .001) among all adolescents from 1999 to 2014. Overall, the prevalence of adolescents with 2 or more risk factors declined, but this decline was only significant for high-income adolescents (44.1%-29.1%, P = .02). CONCLUSIONS: Recent improvements in cardiovascular health have not been equally shared by US adolescents of varying socioeconomic status.

Conversion therapy and suitable timing for subsequent salvage surgery for initially unresectable hepatocellular carcinoma: What is new?

AIM: To review the conversion therapy for initially unresectable hepatocellular carcinoma (HCC) patients and the suitable timing for subsequent salvage surgery. METHODS: A PubMed search was undertaken from 1987 to 2017 to identify articles using the keywords including "unresectable" "hepatocellular carcinoma", "hepatectomy", "conversion therapy", "resection", "salvage surgery" and "downstaging". Additional studies were investigated through a manual search of the references from the articles. The exclusion criteria were duplicates, case reports, case series, videos, contents unrelated to the topic, comments, and editorial essays. The main and widely used conversion therapies and the suitable timing for subsequent salvage surgery were discussed in detail. Two members of our group independently performed the literature search and data extraction. RESULTS: Liver volume measurements [future liver remnant (FLR)/total liver volume or residual liver volume/bodyweight ratio] and function tests (scoring systems and liver stiffness) were often performed in order to justify whether patients were suitable candidates for surgery. Successful conversion therapy was usually defined as downstaging the tumor, increasing FLR and providing subsequent salvage surgery, without increasing complications, morbidity or mortality. The requirements for performing salvage surgery after transcatheter arterial chemoembolization were the achievement of a partial remission in radiology, the disappearance of the portal vein thrombosis, and the lack of extrahepatic metastasis. Patients with a standardized FLR (sFLR) > 20% were good candidates for surgery after portal vein embolization, while other predictive parameters like growth rate, kinetic growth rate were treated as an effective supplementary. There was probably not enough evidence to provide a standard operation time after associating liver partition and portal vein ligation for staged hepatectomy or yttrium-90 microsphere radioembolization. The indications of any combinations of conversion therapies and the subsequent salvage surgery time still need to be carefully and comprehensively evaluated. CONCLUSION: Conversion therapy is recommended for the treatment of initially unresectable HCC, and the suitable subsequent salvage surgery time should be reappraised and is closely related to its previous therapeutic effect.

Overexpression of Bmi­1 promotes epithelial­mesenchymal transition in CD133+Hep G2 cells.

Cancer stem cells (CSCs) and epithelial­mesenchymal transition (EMT) are critical factors contributing to tumor metastasis and recurrence. The BMI1 proto­oncogene (Bmi­1) promotes the development and progression of hematologic malignancies and of several types of solid tumors. The aim of the present study was to explore the mechanism by which Bmi­1 may promote invasion and migration of hepatocellular carcinoma Hep G2 cells. CD133 antigen is a transmembrane glycoprotein and regarded as a cancer stem cells marker in hepatocellular carcinoma. CD133+Hep G2 cells were enriched by magnetic­activated cell sorting and exhibited greater viability compared with CD133­Hep G2 cells, as measured by Cell Counting kit­8 assay. Then, Bmi­1 was overexpressed in CD133+Hep G2 cells by transfection with the Bmi­1/pcDNA3.1(+) expression plasmid, and overexpression was confirmed by reverse­transcription­polymerase chain reaction and western blotting. Overexpression of Bmi­1in CD133+Hep G2 cells resulted in the downregulation of E­cadherin and upregulation of Vimentin at the protein level. The invasion and migration abilities of CD133+Hep G2 cells were increased in the Bmi­1/pcDNA3.1(+)­transfected group, as measured by Transwell invasion and wound healing assays, respectively. In conclusion, Bmi­1 promoted invasion and migration of CD133+Hep G2 cells most likely through inducing EMT. The present findings may offer a potential novel target for the development of hepatocellular carcinoma therapies.

Total and regional adiposity measured by dual-energy X-ray absorptiometry and mortality in NHANES 1999-2006.

OBJECTIVE: Associations of overall and regional body fat measured by dual-energy X-ray absorptiometry with total and cause-specific mortality in the National Health and Nutrition Examination Survey (NHANES) 1999-2006 were investigated. METHODS: This study included 9,471 participants (≥20 years) free of major chronic diseases at baseline. Death information was obtained from the National Death Index (mean follow-up duration: 8.8 years). RESULTS: A total of 682 participants died after 12 months since baseline, with 206 and 170 deaths attributed to cardiovascular diseases (CVD) and cancer, respectively. The second quartile of fat mass percentages (FM%) was used as the reference to account for potential nonlinearity. In the multivariate-adjusted model, participants in the highest quartile of total FM% had increased total mortality; hazard ratio (HR; 95% confidence interval, 95% CI) was 1.48 (1.07-2.04; P < 0.05). Higher total and trunk FM%, but not leg FM%, were significantly associated with an increased CVD mortality; HRs (95% CIs) in the highest quartiles of total, trunk, and leg FM% were 2.24 (1.17-4.31), 1.93 (1.02-3.66), and 1.50 (0.77-2.94), respectively. CONCLUSIONS: Higher total body fat was associated with increased total mortality in U.S. adults. Higher total and trunk fat contents were also associated with increased CVD mortality, although fat accumulation in the lower body was not an independent predictor of mortality.

Bmi-1 promotes the invasion and migration of colon cancer stem cells through the downregulation of E-cadherin.

Metastasis and recurrence are the challenges of cancer therapy. Recently, mounting evidence has suggested that cancer stem cells (CSCs) and epithelial-mesenchymal transition (EMT) are critical factors in tumor metastasis and recurrence. The oncogene, Bmi-1, promotes the development of hematologic malignancies and many solid tumors. The aim of the present study was to elucidate the mechanisms through which Bmi-1 promotes the invasion and migration of colon CSCs (CCSCs) using the HCT116 colon cancer cell line. Sphere formation medium and magnetic­activated cell sorting were used to enrich and screen the CCSCs. CD133 and CD44 were regarded as markers of CCSCs and they were found to be co-expressed in the HCT116 colon cancer cell line. Colony formation assay, cell proliferation assay and viability assay using the Cell Counting Kit-8, and transplantation assay using nude mice injected with CCSCs were used to examine the CCSCs. The CD133+CD44+ HCT116 cells exhibited greater cloning efficiency, an enhanced proliferative ability, increased cell viability and stronger tumorigenicity; these cells were used as the CCSCs for subsequent experiments. In addition, the invasive and migratory abilities of the CD133+CD44+ HCT116 cells were markedly decreased when Bmi-1 was silenced by small interfering RNA (siRNA). The results of RT-qPCR and western blot analysis suggested that Bmi-1 had a negative effect on E-cadherin expression. On the whole, our findings suggest that Bmi-1 promotes the invasion and migration of CCSCs through the downregulation of E-cadherin, possibly by inducing EMT. Our findings thus indicate that Bmi-1 may be a novel therapeutic target for the treatment of colon cancer.

Regulation of activating protein-4-associated metastases of non-small cell lung cancer cells by miR-144.

Activating protein-4 (AP4) has been recently shown to regulate the cancer metastases in some cancers including non-small cell lung cancer (NSCLC). Specifically, AP4 regulates mTor/p21 and transforming growth factor ß (TGFß) receptor signaling pathway to increase an epithelial-mesenchymal transition process to augment cell invasiveness. Nevertheless, how AP4 is regulated in NSCLC has not been studied. Here, we showed that in the specimens from the NSCLC patients, the levels of miR-144 were significantly decreased and the levels of AP4 were significantly increased, compared to the paired non-tumor lung tissue. The levels of miR-144 and AP4 inversely correlated in patients' specimens. Bioinformatics analyses revealed that miR-144 targeted the 3'-UTR of AP4 mRNA to inhibit its translation, confirmed by luciferase-reporter assay. Moreover, miR-144 overexpression inhibited AP4-mediated cell invasiveness, while miR-144 depletion increased AP4-mediated cell invasiveness in NSCLC cells. Together, our data suggest that miR-144 suppression may be the cause of the increased levels of AP4, as well as the augmented cancer metastases, in NSCLC.