Construction of molecular subgroups of ulcerative colitis.

OBJECTIVE: Ulcerative colitis (UC), a chronic inflammatory disease of the colon with unknown etiology, is characterized by remission and recurrence. At present, a considerable number of UC cases are misdiagnosed or delayed in diagnosis and treatment. We aimed to identify UC-related genes to aid the development of drugs for this condition. PATIENTS AND METHODS: Transcriptome data of 362 patients with UC and 126 control subjects were obtained from the Gene Expression Omnibus. The 362 patients with UC were subgrouped using unsupervised machine learning. R software was used to analyze the clinical characteristics of the subgroups, screen subgroup-specific genes, assess the relationships between gene modules and clinical characteristics using weighted gene co-expression network analysis, and perform Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the subgroups. RESULTS: Patients with UC were classified into two subgroups. Genes specific to subgroup I included IL21R, ATP8B2, and PLEKHO1. Severe disease tended to be associated with immune cell infiltration; anti-tumor necrosis factor (TNF)-α antibodies and ustekinumab may have been effective in this subgroup. Subgroup II-specific genes included SLC4A4, EPB41L4B, and PLCE1. Patients in this subgroup had mild clinical conditions; however, their disease was more likely to progress to colorectal cancer. Thus, 5-aminosalicylic acid-based drugs may be effective for the treatment of UC in these patients. CONCLUSIONS: We divided UC into two molecular subgroups based on transcriptome data, providing molecular evidence for the development of diagnostic methods and individualized treatment strategies for UC.

A new prognostic model for brain metastases of specific primary tumors with stereotactic radiotherapy.

PURPOSE: Stereotactic radiotherapy (SRT) was widely used in brain metastases (BM), especially in oligometastases. It is imperative to develop a new prognostic score to predict the overall survival (OS) of brain metastases based on prognostic factors for specific primary tumors. MATERIAL AND METHOD: One hundred and ninety-seven patients were involved in the training cohort to develop a new prognostic score to predict the overall survival (OS) of brain metastases for specific primary tumors. Independent prognostic factors were confirmed using a Cox regression model. The score was developed based on clinical prognostic factors of OS with Cox proportional hazards model. The result was validated in another cohort with 56 participants to evaluate the performance of the score. RESULTS: One hundred and ninety-seven patients with 329 brain metastases received SRT. For NSCLC, the significant prognostic factors were extracranial metastases, target therapy and number of brain metastases. For gastrointestinal cancer, the significant prognostic factors were target therapy and number of brain metastases. CONCLUSION: The prognostic factors scores were varied by the histologic types which can be used to efficiently stratify for selected patients with brain-metastasis.

Adenosine diphosphate-ribosylation factor-like 15 can regulate glycolysis and lipogenesis related genes in colon cancer.

This study was designed to investigate the potential key genes of ADP-ribosylation factor-like 15 (ARL15) regulating glycolysis and lipogenesis in colon cancer. Hematoxylin-eosin (HE) staining and immunohistochemistry were used to observe the expression of ARL15 in 10 normal colon tissues and 10 colon cancer tissues. Immunofluorescence staining was used to observe the expression position of ARL15 in normal human colorectal mucosa cells (FHC) and colon cancer cells (HCT116 and SW620) with a confocal microscope. The ARL15 plasmid and small interfering RNA (siRNA) were constructed. After transfection, the expression levels of glycolysis and lipogenesis regulatory enzymes and messenger RNA (mRNA) transcription of ARL15 in over-expressed and silenced colon cancer cells were detected by Western blotting and real-time quantitative PCR (qRT-PCR). High expression of ARL15 in colon cancer tissue and low expression in normal colon tissue, and all expression are in the cytosol. The expression position of ARL15 in the FHC, HCT116, and SW620 cells was consistent and mainly distributed in the cytosol. After the pCMV-3Tag-2-ARL15 plasmid was transfected in HCT116, the protein expressions of FASN, AKT, P-AKT, P-GSK, SREBP-1 (p125) (p<0.01), and AMPK (p<0.001) were higher than those in the control group. The mRNA transcription level of FASN, GSK, AMPKa1, and SREBP-1 gene was higher than control group after the over-expression of ARL15. After the ARL15-siRNA was transfected in HCT116, the protein expression levels of PKM2, PFK, FASN, AKT, P-AKT, P-GSK, and AMPK decreased compared with the control group, (p<0.05). The mRNA transcription level of FASN, GSK, AMPKα1 gene was lower than control group after the low-expression of ARL15 (p<0.05). After adding 2 µM JIB-04, ARL15 in HCT116 showed statistical differences compared with the control group at 12 h, 24 h and 36 h (p<0.05). After adding 2 µM JIB-04, the protein expression levels of AKT, p-GSK, FASN, AMPK and SREBP-1 in HCT116 cells decreased significantly after 24 h. It was also found that the expression levels of AKT, P-GSK, FASN, AMPK and SREBP-1 genes in the dose-adding group were significantly lower than those in the control group. In summary, ARL15 may promote the occurrence of colon cancer by increasing the expression of protein kinase B/AMP-activated protein kinase (AKT/AMPK) downstream regulatory enzymes for glycogenesis and lipogenesis. JIB-04 can target ARL15 and affect its expression as well as the expressions of glucose and lipid metabolity-related proteins in AKT and AMPK signaling pathways.

Impact of insulin use on outcomes of diabetic breast cancer patients: a systematic review and meta-analysis.

OBJECTIVE: The study aimed to assess the impact of insulin use on outcomes of breast cancer patients with diabetes mellitus (DM). MATERIALS AND METHODS: Databases of PubMed, Embase, and CENTRAL were searched to identify all types of studies comparing mortality or recurrence between insulin and non-insulin DM patients with breast cancer. Adjusted hazard ratios (HR) were pooled for a meta-analysis. RESULTS: Eleven studies were included. Meta-analysis indicated a statistically significant increased risk of all-cause mortality in insulin users as compared to non-users (HR: 1.52 95% CI: 1.23 to 1.86 I2=83% p<0.0001). Our results also demonstrated a statistically significant increase in the risk of breast cancer mortality amongst insulin users as compared to non-users (HR: 1.33 95% CI: 1.08 to 1.63 I2=43% p=0.007). Only four studies assessed the impact of insulin therapy on recurrence rates. Meta-analysis indicated a statistically significant increased risk of breast cancer recurrence in insulin users vs. non-users (HR: 1.43 95% CI: 1.13 to 1.80 I2=0% p=0.003). Mortality results were stable on sensitivity analysis. CONCLUSIONS: Diabetic breast cancer patients on insulin have increased mortality and recurrence rates as compared to insulin non-users. Owing to the several limitations of the review, results should be interpreted with caution. Future studies should assess the impact of timing, duration, dosage, and type of insulin therapy on clinical outcomes.

Pharmacological characterization of three chicken melanocortin-3 receptor mutants.

The melanocortin-3 receptor (MC3R) is a G protein-coupled receptor and potentially important in production traits. Three naturally occurring mutations (M54L, G104S, and L151R) in chicken MC3R (cMC3R) were reported previously to be associated with production traits. Here, we inserted the full-length cMC3R coding sequence into pcDNA3.1(+) and generated the 3 mutations by site-directed mutagenesis. The total and cell surface expression of the receptors was measured by flow cytometry. We analyzed the pharmacological characteristics, including binding and cyclic adenosine monophosphate (cAMP) and mitogen-activated protein kinase (MAPK) signaling, using 6 ligands ([Nle4, D-Phe7]-α-melanocyte stimulating hormone (MSH), α-, ß-, γ-, and D-Trp8-γ-MSHs, and agouti-related peptide). All mutants had similar total and cell surface expression as the wild-type (WT) cMC3R. M54L had similar pharmacological properties as the WT cMC3R. G104S did not exhibit any specific binding but had minimal response to α-, ß-, γ-, and D-Trp8-γ-MSH, although it generated 24% WT response when stimulated by NDP-MSH. Although L151R had normal binding, the responses to agonists were reduced to approximately 25% of that of the WT. In MAPK signaling, all 3 mutants showed significantly increased agonist-stimulated phosphorylation of extracellular signal-regulated protein kinases 1/2, indicating the existence of biased signaling at G104S and L151R. In summary, our studies demonstrated that although all 3 mutations are significantly associated with production traits, only G104S and L151R had severe defects in receptor pharmacology. How M54L might cause production trait differences remains to be investigated.

Expression levels of plasma miRNA-21 and NT-proBNP in children with Kawasaki disease and their clinical significance.

OBJECTIVE: The purpose of this study was to detect the expression levels of plasma microRNA-21 (miRNA-21) and NT-proBNP (N-terminal prohormone of brain natriuretic peptide) in children with Kawasaki disease (KD), as well as their clinical significance. PATIENTS AND METHODS: Children with KD (n=100) who were treated in our hospital from June 2017 to May 2019 were included. In the same period, non-KD children with febrile diseases were included as controls. Plasma levels of miRNA-21 and NT-proBNP were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and electrochemiluminescence, respectively. Then, the relationship between miRNA-21 and NT-proBNP in children with KD was analyzed by Pearson's correlation test. Potential factors influencing KD were analyzed by Multivariate logistic regression test. Finally, receiver operating characteristic (ROC) curves were depicted to assess the diagnostic potentials of miRNA-21 and NT-proBNP in KD. RESULTS: The results showed that miRNA-21 and NT-proBNP levels were higher in children with KD. Plasma level of miRNA-21 was positively correlated with NT-proBNP level in children with KD. Besides, both miRNA-21 and NT-proBNP were risk factors influencing the onset of KD. According to ROC curves, the sensitivity and specificity of miRNA-21 in diagnosing KD was 83% and 89%, respectively (AUC=0.9212, 95% CI: 0.8809-0.9614, cut-off value=1.985). NT-proBNP also displayed diagnostic potential in KD (AUC=0.9788, 95% CI: 0.9630-0.9946, cut-off value=265.6, sensitivity=88%, specificity=95%). CONCLUSIONS: MiRNA-21 and NT-proBNP are upregulated in plasma of children with KD. They are positively correlated with each other and serve as risk factors for KD. Both of them can be utilized as indicators of auxiliary diagnosis in KD.

Characterization of antibody and memory T-cell response in H7N9 survivors: a cross-sectional analysis.

OBJECTIVES: Despite the importance of immunological memory for protective immunity against viral infection, whether H7N9-specific antibodies and memory T-cell responses remain detectable years after the original infection is unknown. METHODS: A cross-sectional study was conducted to investigate the immune memory responses of H7N9 patients who contracted the disease and survived during the 2013-2016 epidemics in China. Sustainability of antibodies and T-cell memory to H7N9 virus were examined. Healthy individuals receiving routine medical examinations in a physical examination centre were recruited as control. RESULTS: A total of 75 survivors were enrolled and classified into four groups based on the time elapsed from illness onset to specimen collection: 3 months (n = 14), 14 months (n = 14), 26 months (n = 28) and 36 months (n = 19). Approximately 36 months after infection, the geometric mean titres of virus-specific antibodies were significantly lower than titres in patients 3 months after infection, but 16 of 19 (84.2%) survivors in the 36-month interval had microneutralization (MN) titres ≥40. Despite the overall declining trend, the percentages of virus-specific cytokine-secreting memory CD4+ and CD8+ T cells remained higher in survivors at nearly all time-points in comparison with control individuals. Linear regression analysis showed that severe disease (mean titre ratio 2.77, 95% CI 1.17-6.49) was associated with higher haemagglutination inhibition (HI) titre and female sex for both HI (1.92, 1.02-3.57) and MN (3.33, 1.26-9.09) antibody, whereas female sex (mean percentage ratio 1.69, 95% CI 1.08-2.63), underlying medical conditions (1.94, 95% CI 1.09-3.46) and lack of antiviral therapy (2.08, 95% CI 1.04-4.17) were predictors for higher T-cell responses. CONCLUSIONS: Survivors of H7N9 virus infection produced long-term antibodies and memory T-cell responses. Our findings warrant further serological investigation in general and high-risk populations and have important implications for vaccine design and development.

LncRNA MALAT1 knockdown alleviates myocardial apoptosis in rats with myocardial ischemia-reperfusion through activating PI3K/AKT signaling pathway.

OBJECTIVE: To observe the effect of long non-coding ribonucleic acid metastasis-associated lung adenocarcinoma transcript 1 (lncRNA MALAT1) on the myocardial ischemia-reperfusion (I/R) injury in rats and to explore its potential mechanism, to provide certain references for clinical prevention and treatment of myocardial I/R injury. MATERIALS AND METHODS: A total of 60 male Wistar rats were randomly divided into the Control group (n=20), I/R group (n=20) and I/R + MALAT1 small-interfering RNA (siRNA) group (n=20) using a random number table. The I/R model was established through recanalization after ligation of left anterior descending coronary artery (LAD), and the MALAT1 knockdown model was established via tail intravenous injection of MALAT1 siRNA in the I/R + MALAT1 siRNA group. The ejection fraction (EF%) and fractional shortening (FS%) of rats in each group were detected via echocardiography and the infarction area in each group was detected using 2,3,5-triphenyl tetrazolium chloride (TTC) assay. Moreover, the morphological changes in myocardial cells in each group were detected via hematoxylin-eosin (H&E) staining, and the myocardial apoptosis level was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. At the same time, the expression levels of the anti-apoptotic protein B-cell lymphoma-2 (Bcl-2) and pro-apoptotic protein Bcl-2 associated X protein (Bax) in myocardial tissues in each group were determined via Western blotting. Finally, the effect of MALAT1 knockdown on the phosphatidylinositol 3-hydroxy kinase/protein kinase B (PI3K/AKT) protein expression was detected via Western blotting. RESULTS: The expression level of lncRNA MALAT1 in myocardial tissues was significantly higher in the I/R group than that in the Control group (p<0.05). The MALAT1 knockdown could significantly improve the cardiac insufficiency caused by I/R injury, and increase both EF% and FS% in rats (p<0.05). In addition, the MALAT1 knockdown could markedly inhibit myocardial infarction caused by I/R injury and reduce the infarction area from (62.12 ± 1.29) to (27.66 ± 3.58; p<0.05). The results of the H&E staining showed that the myofilaments were arranged more orderly, the degrees of degradation and necrosis were lower and the cellular edema was significantly alleviated in the I/R + MALAT1 siRNA group compared with those in the I/R group. According to the results of TUNEL staining, the rats in I/R + MALAT1 siRNA group had a markedly lower level of myocardial apoptosis than the I/R group (p<0.05), and the Bax/Bcl-2 ratio also remarkably declined in the I/R + MALAT1 siRNA group (p<0.05). Furthermore, the results of Western blotting revealed that MALAT1 siRNA could significantly reverse the I/R injury-induced inhibition on the AKT phosphorylation (p<0.05). CONCLUSIONS: The MALAT1 knockdown can markedly improve the I/R-induced myocardial injury and promote the cardiac function of rats, whose mechanism may be related to the activation of the AKT signaling pathway by MALAT1 siRNA. Therefore, lncRNA MALAT1 is expected to be a new therapeutic target for myocardial I/R injury.

Down-regulation of miR-1236-3p is correlated with clinical progression and unfavorable prognosis in gastric cancer.

OBJECTIVE: MiRNAs have been considered to participate in many processes of various cancers, including gastric cancer (GC). However, the significance of miRNAs in the progression prognosis of GC is largely unknown. The goal of this study was to determine the expression level of miR-1236-3p in GC and its clinical association. PATIENTS AND METHODS: Clinical specimens from GC patients were obtained to quantify the expression level of miR-1236-3p using quantitative Real-time PCR. The correlation between the miR-1236-3p levels and the clinicopathological factors of the GC patients was analyzed. The association between miR-1236-3p expression and overall survival was estimated by the Kaplan-Meier method. The significance of different variables with respect to survival was analyzed using the univariate and multivariate Cox proportional hazards model. RESULTS: We found that miR-1236-3p was significantly downregulated in GC tissues compared to non-tumor gastric tissues (p < 0.01). The levels of miR-1236-3p expression were associated significantly with clinical stage (p = 0.002), lymph node metastasis (p = 0.000) and distant metastasis (p = 0.017). Kaplan-Meier survival analysis showed that patients in the high miR-1236-3p expression group had better overall survival than those in the low miR-1236-3p expression group (p = 0.0039). Moreover, we confirmed that miR-1236-3p was an independent poor prognostic factor for GC patients through univariate and multivariate analysis. CONCLUSIONS: Our findings provide evidence that miR-1236-3p expression is frequently decreased in GC tissues, and its low expression may be a significant prognostic factor for poor survival in GC patients.

Short-term combined treatment with exenatide and metformin is superior to glimepiride combined metformin in improvement of serum testosterone levels in type 2 diabetic patients with obesity.

Men with type 2 diabetes (T2D) and obesity are often characterised by low testosterone (T). We aimed to determine whether exenatide (EXE) combined metformin (MET) treatment has a better effect on serum total testosterone (TT) levels than glimepiride (GLI) combined MET treatment in men with T2D and obesity. In a multicentre, 12-week observational study, 176 obese T2D men with failed glycaemic control were included in the study: ninety men (mean age, 43.00 ± 8.50 years) in EXE + MET group and 86 men (mean age, 44.00 ± 7.00 years) in GLI + MET group. Serum TT levels were more significantly increased in EXE + MET group than GLI + MET group (121.72 ± 56.73 ng/dl versus 34.67 ± 16.30 ng/dl). The increasement of TT levels in those patients who lost body weight ≥5% was significantly greater than those who lost weight <5% in the two groups. The changes in TT levels are closely related to the changes in waist circumference (r = -.443, p < .001). Sexual function assessment of EXE + MET group was more significantly improved than GLI + MET group (p < .001). No serious adverse events were observed. In conclusion, short-term combined treatment with EXE and MET is superior to GLI combined MET treatment in the improvement of serum TT levels, which could lead to an improvement of sexual hypofunction in patients with obesity and T2D.

Relationship between the flavonoid composition and flower colour variation in Victoria.

Victoria (Nymphaeaceae), an annual or perennial aquatic plant genus, contains only two species: V. amazonica (Poepp.) J. C. Sowerby and V. cruziana A. D. Orb. Both species have large floating leaves and variable flower colour. Both Victoria species are night bloomers, which have white petals on the first blooming night that then turn pink or ruby red on the second blooming day. The mechanism of the colour change of Victoria petals during anthesis is still unclear. In this study, flavonoids in Victoria petals of both species were evaluated and quantified by high-performance liquid chromatography with photodiode array detection (HPLC-DAD) and by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) for the first time. In total, 14 flavonoids were detected in Victoria petals, including 4 anthocyanins and 10 flavonols. The flavonoid compositions differed across the two species, resulting in different colours between the inner and outer petals. With increased anthocyanin content across blooming days, the colour of Victoria flowers changed over time. The results of this study will improve understanding of the chemical mechanism of colour formation and lay the foundation for selective colour breeding in Victoria.

MiR-214 protects MC3T3-E1 osteoblasts against H2O2-induced apoptosis by suppressing oxidative stress and targeting ATF4.

OBJECTIVE: Fragility fracture is one of the common complications of osteoporosis. Elevated oxidative stress-induced apoptosis is thought to be one of the unfavorable factors to osteoblastic dysfunction, which increased the risk of bone fracture. However, the molecular mechanisms for oxidative stress-induced osteoblast cells apoptosis still need to be elucidated. This study aims to investigate the protective function of miR-214 in H2O2-induced apoptosis of MC3T3-E1 osteoblasts. MATERIALS AND METHODS: MC3T3-E1 cells were treated with 400 µM H2O2. Flow cytometry was adopted to detect the apoptosis rate; malondialdehyde (MDA) and glutathione peroxidase (Gpx) levels were used to determine the reactive oxygen species (ROS) level. Reverse transcription-polymerase chain reaction (RT-PCR) was employed to test the expression level of miR-214 and ATF4. After transfected MC3T3-E1 cells with miR-214 mimics and inhibitor, RT-PCR was used to detect activating transcription factor 4 (ATF4) expression level. RESULTS: H2O2 treatment increased ROS induced intracellular oxidative injury. Flow cytometry showed that 400 µM H2O2 induced the apoptosis of MC3T3-E1 cells. Moreover, RT-PCR showed decreased expression level of MiR-214. Furthermore, the apoptosis induced by high ROS level was reversed by increased miR-214 expression level. The regulatory ability of MiR-214 to apoptosis is by regulating ATF4 expression. CONCLUSIONS: miR-214 plays a protective role in H2O2 induced MC3T3 osteoblasts apoptosis and its protective effect is proceeded by regulating ROS level and ATF4 expression level.

The motility of esophageal sphincters during liquid and solid bolus swallows: a multicenter normative value study of high-resolution manometry in China.

BACKGROUND: It is gradually accepted that solid bolus swallow needs to be added to the procedure of manometry. The motility differences in the upper esophageal sphincter (UES) and lower esophageal sphincter (LES) were not well described. Sierra Scientific Instruments solid-state high-resolution manometry (HRM) system, the most popular HRM system in China, lacks the Chinese normative values for both liquid and solid bolus swallow parameters. METHODS: The esophageal HRM data of 88 healthy volunteers were analyzed. The parameters of both sphincters in resting stage were summarized and those during solid and liquid swallows were compared. KEY RESULTS: Normative HRM values of sphincter parameters in solid and liquid bolus swallows in China were established. The UES residual pressure of solid bolus swallows was lower than that of liquid bolus (0.3±5.5 mm Hg vs 4.8±5.9 mm Hg, P=.000). The time parameters of UES relaxation between two types of bolus swallows were similar. In solid bolus swallows, the intrabolus pressure (IBP) (13.8±5.1 mm Hg vs 10.9±5.7 mm Hg, P=.000) and LES relaxation time (11.0±2.1 seconds vs 8.7±1.3 seconds, P=.000) were higher. The 4-second integrated relaxation pressure between both bolus swallows was similar. CONCLUSIONS & INFERENCES: The function of the UES and LES between solid and liquid bolus swallows is different. Chinese HRM parameters are different from the Chicago Classification (http://www.chictr.org.cn, Number ChiCTR-EOC-15007147).

Low miR-133a expression is a predictor of outcome in patients with esophageal squamous cell cancer.

OBJECTIVE: Identification and development of new biomarkers could be beneficial for diagnosis and prognosis of esophageal squamous cell cancer (ESCC) patients. The aim of this study was to examine the clinical significance of miR-133a expression in tissues from ESCC patients. PATIENTS AND METHODS: Quantitative real-time reverse transcriptive-PCR (qRT-PCR) was performed to examine miR-133a expression levels in 126 ESCC tissues. Survival curves were plotted using the Kaplan-Meier method and differences in survival rates were analyzed using the log-rank test. Univariate and multivariate Cox regression analyses were conducted to determine whether miR-133a was an independent predictor of survival. RESULTS: We found that the levels of miR-133a in ESCC tissues exhibited lower than that in matched normal tissues (p < 0.01). Low miR-133a expression was positively associated with T stage of ESCC (p = 0.013) and tumor length (p = 0.007). Moreover, low levels of miR-133a was associated with lower overall survival (OS) (p = 0.001) and disease-free survival (DFS) (p = 0.001). According to multivariate analysis, miR-133a level was confirmed to be an independent prognostic factor for worse survival. CONCLUSIONS: MiR-133a may represent a prognostic biomarker and therapeutic target in esophageal squamous cell cancer prognosis and treatment.

MicroRNA-183 promotes proliferation and invasion in oesophageal squamous cell carcinoma by targeting programmed cell death 4.

BACKGROUND: Dysregulated microRNAs (miRNAs) can serve as oncogenes or suppressors and are associated with many cancers, including oesophageal squamous cell carcinoma (ESCC). METHODS: An alignment miRNA array was used to identify differentially expressed miRNAs in ESCC tissues. The expression of miR-183 and programmed cell death 4 (PDCD4) in oesophageal tissues from ESCC and early oesophageal carcinoma patients was examined by quantitative reverse transcriptase PCR and western blotting. A luciferase assay was performed to confirm miR-183 target genes. The effects of miR-183 on ESCC cells and the associated mechanisms were established by in vitro experiments. RESULTS: We identified 51 upregulated miRNAs and 17 downregulated miRNAs in our array, and miR-183 was one of the most upregulated miRNAs. An inverse correlation between miR-183 and PDCD4 levels was found in ESCC tissues. Upregulated expression of miR-183 was not correlated with tumour stage or lymphatic metastasis in ESCC patients. The luciferase assay confirmed that miR-183 directly interacted with the PDCD4 mRNA 3'-untranslated region in ESCC cells. Overexpression of miR-183 led to decreased PDCD4 protein levels and promoted ESCC cell proliferation and invasion. Inhibition of the PI3K/Akt signalling pathway increased PDCD4 protein levels and decreased miR-183 expression in ESCC cells. CONCLUSIONS: MiR-183 promotes ESCC cell proliferation and invasion by directly targeting PDCD4, which suggests that it is involved in the pathogenesis of ESCC.

Heroin abuse accelerates biological aging: a novel insight from telomerase and brain imaging interaction.

Heroin abuse and natural aging exert common influences on immunological cell functioning. This observation led to a recent and untested idea that aging may be accelerated in abusers of heroin. We examined this claim by testing whether heroin use is associated with premature aging at both cellular and brain system levels. A group of abstinent heroin users (n=33) and matched healthy controls (n=30) were recruited and measured on various biological indicators of aging. These measures included peripheral blood telomerase activity, which reflects cellular aging, and both structural and functional measures of brain magnetic resonance imaging. We found that heroin users were characterized by significantly low telomerase activity (0.21 vs 1.78; 88% reduction; t(61)=6.96, P<0.001; 95% confidence interval=1.12-2.02), which interacted with heroin use to affect the structural integrity of gray and white matter of the prefrontal cortex (PFC; AlphaSim corrected P<0.05), a key brain region implicated in aging. Using the PFC location identified from the structural analyses as a 'seed' region, it was further revealed that telomerase activity interacted with heroin use to impact age-sensitive brain functional networks (AlphaSim corrected P<0.05), which correlated with behavioral performance on executive functioning, memory and attentional control (Pearson correlation, all P<0.05). To our knowledge, this study is the first to attempt a direct integration of peripheral molecular, brain system and behavioral measures in the context of substance abuse. The present finding that heroin abuse is associated with accelerated aging at both cellular and brain system levels is novel and forms a unique contribution to our knowledge in how the biological processes of drug abusers may be disrupted.

First Report of Colletotrichum acutatum Associated with Stem Blight of Blueberry Plants in China.

An anthracnose disease was observed on stems of high-bush blueberry plants (Vaccinium corymbosum L.) in Liaoning Province, China in 2012. The typical symptoms consist of sudden wilting and dieback of stems during the growing season. Dark brown lesions originate from infected buds and kill portions of the stems. Lesions have grayish white centers, with the necrotic areas becoming 6 to 8 cm in length. Disinfected stem pieces were placed on potato dextrose agar (PDA) and incubated at 28°C for 5 to 7 days, after which the emerging colonies were transferred to fresh PDA. All isolates initially produced white growth, but turned pink after 7 days before becoming blackish green. The average colony diameter was 65.5 to 75.0 mm after 7 days. Conidia were aseptate, hyaline, fusiform to ellipsoid, 8.5 to 16.5 × 2.5 to 4.0 µm in size and single celled with two to seven oil globules. Setae were not found on the acervuli. These characteristics matched published descriptions of Colletotrichum acutatum (1) (teleomorph Glomerella acutata). Pathogenicity test was confirmed in 15 2-year-old healthy potted plants of cv. Berkeley. Stems of 10 plants were punctured with flamed needles and sprayed with 5 ml of conidial suspension (106 conidia per ml in sterile distilled water) of isolate LNSW1. Five control plants were inoculated with sterile distilled water. Seven days after inoculation, eight of the 10 blueberry plants exhibited stem lesions, leaf chlorosis, followed by branch dieback 15 days post-inoculation. The symptoms were similar to those observed on diseased plants in the field, and no lesions were observed on control plants. The pathogen was reisolated from the margin of lesions and identified by colony growth characteristics on PDA. PCR amplification of one isolate (LNSW1) was carried out by utilizing the universal rDNA-ITS primer pair ITS1/ITS4. The sequence (557 bp) of isolate LNSW1 (GenBank Accession No. JX392857) showed 99% identity to G. acutata (AB443950) and C. acutatum (AJ749672) in a BLAST search. An approximately 490-bp fragment was amplified from LNSW1 by the species-specific primer pair CaInt2/ITS4 (2). The pathogen was identified as G. acutata (asexual stage: C. acutatum J.H. Simmonds) on the basis of morphological characters, rDNA-ITS sequence analysis, and a PCR product with species-specific primers. To our knowledge, this is the first report of C. acutatum in high-bush blueberry plants in China. References: (1) C. Lei et al. Fungal Diversity 12:183, 2009. (2) S. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996.

Effects of breed and dietary nutrient density on the growth performance, blood metabolite, and genes expression of target of rapamycin (TOR) signalling pathway of female broiler chickens.

This study was conducted to compare the effects of exchanged diets with identical energy level on characteristics of slow-growing (WENs Yellow-Feathered Chicken, WYFC) and fast-growing (White Recessive Rock Chicken, WRRC) female chickens. A total of 1450 WYFC and 1150 WRRC 1-day-old female hatchlings were used. A high-nutrient-density (HND) diet and a low-nutrient-density (LND) diet were formulated for three phases. A completely randomized experimental design with a 2 × 2 factorial arrangement (diet and breed), each with five replicates of 145 and 115 birds, was applied. The results showed that WRRC had a higher body weight (BW), average daily feed intake and average daily gain than WYFC throughout the experiment (p<.05). WYFC that were provided with HND groups had a higher BW only in the starter and grower phases, whereas WRRC had a higher BW in the HND group than in LND groups throughout the experiment. The feed:gain ratio and protein efficiency ratio (PER) were better for WRRC in the starter and grower phases; however, these ratios were better for WYFC in the finisher period. The LND groups had a higher PER throughout the experiment for both breeds (p<0.05). The breast and leg muscle weights were higher for WRRC compared with WYFC during the grower and finisher phases (p<0.05). WRRC had a lower liver index but higher serum UA and alkaline phosphatase (ALP) concentrations than WYFC (p<0.05). No diet effect was observed on organ indices, muscle yields or blood responses. The gene expressions of Rheb, TOR, S6K1 and 4E-BP1 in gastrocnemius muscle were the highest in the WYFC-LND groups at 63 and 105 days (p<0.05). These findings suggested that different genotypes respond differently to changes in dietary nutrient density and that lower-nutrient-density diets are optimal for the long-term housing of broiler chickens.