Mixture of polysaccharide and nucleic acid extracted from Bacillus Calmette-Guerin (BCG) enhances immune response of infectious bursal disease virus vaccine in chickens.

In this study, the immune response induced by a mixture of polysaccharide and nucleic acid extracted from Bacillus Calmette-Guerin (BCG) was evaluated in chickens inoculated with infectious bursal disease virus (IBDV) vaccine. After the mixture was injected intramuscularly at a dose of 0.075, 0.15 or 0.3 mg·kg(-1)·day(-1) for 3 days, the 14-day-old chickens were inoculated with the attenuated IBDV vaccine via intranasal and ocular routes. The relative weight of bursa of Fabricius (BF) and thymus, the serum IBD antibody titer, the CD4+/CD8+ ratio, and the concentrations of IFN-γ, IL-2 and IL-6 in peripheral blood were investigated on days 5, 15 and 25. The IBD antibody titer in BCG-treated groups was higher than in the negative control and only IBD-vaccinated chickens, indicating that the mixture of BCG can significantly enhance chicken humoral response. CD4+/CD8+ and the secretions of IFN-γ, IL-2 and IL-6 were also clearly increased compared with that in the negative control and IBD-vaccinated chickens, indicating that the mixture can also enhance the cell-mediated immune response. The results also showed that the relative weights of BF and thymus increased after chickens were inoculated with BCG, indicating that the BCG mixture can clearly enhance the immunity of IBD-vaccine and can be expected to be viewed as a candidate for a new type of immune adjuvant.

Protective effect of interleukin-10 and recombinant human keratinocyte growth factor-2 on ventilation-induced lung injury in rats.

A rat model of ventilation-induced lung injury (VILI) during anesthesia was generated to investigate the potential role and possible mechanism of interleukin-10 (IL-10) and recombinant human keratinocyte growth factor-2 (rhKGF-2) in protecting anesthetized rats against VILI. A total of 50 male SD rats were randomly divided into 5 groups (N = 10 each): control, VILI, IL-10, rhKGF-2, and IL-10 + rhKGF-2. The VILI (model) group was generated via ventilation, with a tidal volume of 20 mL/kg. Rats in the IL-10 and rhKGF-2 groups received 8 mg/kg IL-10 and 5 mg/kg rhKGF-2, respectively, prior to ventilation. The rats in the IL-10 + rhKGF-2 group received both 8 mg/kg IL-10 and 5 mg/kg rhKGF-2 72 h before ventilation. The total number of nucleated cells and neutrophils in the bronchial alveolar lavage fluid was quantified, and the pathological changes in the pulmonary tissues examined by hematoxylin and eosin staining. The transcript and protein levels of surfactant protein C (SP-C) in lung tissues were detected by real-time polymerase chain reaction and western blot analyses. The SP-C mRNA expression in both IL-10 and rhKGF-2 groups was similar to that in the VILI group. However, this was significantly elevated in the combined treatment group (P < 0.05), indicating that IL-10 and rhKGF-2 could synergistically protect the lung tissue from VILI via the enhancement of SP-C mRNA expression in lung tissues. The protein assay showed a decreased level of infiltration and activation of inflammatory cells, in addition to increased expression of SP-C, thereby confirming the efficacy of this treatment in preventing VILI during anesthesia.

Interference on cytoplasmic polyadenylation element-binding proteins affects the invasion ability of glioma stem cells.

Glioma stem cells derived from primary cultures were divided into an experiment group, a control group, and a blank group and subjected to cytoplasmic polyadenilation element-binding protein (CPEBs) interference, transfection with empty vector, and normal culture, respectively, to compare their invasion abilities. Western blotting showed that siRNA-3 had the strongest interfering effect on CPEBs. CPEBs were expressed in the experiment group with green fluorescence at an expression rate of over 70%. Significantly lower CPEB expression was observed in the experiment group compared to in the control and blank groups (P < 0.05). After 48-h treatment, the apoptotic rate in the experiment group was 21.43%, which was significantly higher than that in the blank (0.51%) and control (1.43%) groups (P < 0.05). After 3 days of treatment, the experiment group grew significantly more slowly than did the control and blank groups (P < 0.05). The transwell invasion assay showed that significantly fewer cells in the experiment group penetrated the membrane than did cells in the control and blank groups (P < 0.05). After CPEB interference, the growth, proliferation, and invasion of glioma stem cells were substantially inhibited, providing support for targeted therapy of glioma and for improving prognosis.

Investigation of the value of miR-21 in the diagnosis of early stage HCC and its prognosis: a meta-analysis.

The diagnostic and prognostic value of miR-21 has been examined for hepatocellular carcinoma (HCC), with inconsistent results. Present meta-analysis summarized the diagnostic accuracy and the predictive role for survival of miR-21 in patients with HCC. All eligible studies were searched using PubMed, EMBASE, and Chinese National Knowledge Infrastructure (CNKI) databases up to October 2014. For the diagnostic meta-analysis, the indices of miR-21 in the diagnosis of HCC were pooled using bivariate random-effect approach models. For the prognostic meta-analysis, data were synthesized with a random effect model, and the hazard ratio (HR) or odd ratio (OR) with its 95% confidence interval (95%CI) was used as the effect size estimate. Ten studies dealing with HCC were included. The overall pooled results for sensitivity, specificity, and the area under the curve (AUC) for the diagnostic meta-analysis (five studies) were 74.0 (95%CI = 61.0-85.0), 78.0 (95%CI = 67.0-86.0), and 0.83 (95%CI = 0.80-0.86), respectively. The combined data for the prognostic meta-analysis (seven studies) suggested that miR-21 overexpression in HCC correlated with poor overall survival [HR = 1.19 (95%CI = 0.44-1.94)], and higher miR-21 expression was associated with tumor, node, metastases (TNM) stage [OR = 0.34 (95%CI = 0.13-0.91)]. We concluded that miR-21 might be complementary to alpha fetal protein in HCC diagnosis, and might serve as an attractive estimator of HCC. We also demonstrated that miR-21 overexpression was associated with HCC TNM stage and with poor survival. As our study was limited, additional prospective studies are needed to validate these results.

Expression and clinical significance of IMP3 in microdissected premalignant and malignant pancreatic lesions.

INTRODUCTION: Insulin-like growth factor 2 (IGF-2) mRNA-binding protein 3 (IMP3) is overexpressed in pancreatic cancer, while remaining undetectable in the normal pancreas, indicating its important role in pancreatic cancer pathogenesis. The role of IMP3 in pancreatic carcinogenesis has not been fully understood. The main goal of this study was to probe the expression profile of IMP3 in different stages of pancreatic ductal adenocarcinoma (PDAC) development, and evaluate their prognostic significance in PDAC patients. MATERIALS AND METHODS: We used quantitative real-time RT-PCR combined manual microdissection to precisely detect IMP3 expression in 97 microdissected foci from 50 patients with PDAC. Nonparametric test, Log-rank test and Cox regression analysis were used to evaluate the clinical significance of DNMTs expression. RESULTS: Expression of IMP3 increased from normal duct to pancreatic intraductal neoplasia and to PDAC. IMP3 mRNA expression statistically correlated with TNM staging. Univariate analysis showed that high level of IMP3 expression, tumor differentiation, TNM staging and alcohol consumption were statistically significant risk factors. Multivariate analysis showed that high level of IMP3 expression and tumor differentiation were statistically significant independent poor prognostic factors. CONCLUSIONS: These results suggested that pancreatic carcinogenesis involves an increased IMP3 mRNA expression, and it may become valuable diagnostic and prognostic markers as well as potential therapeutic targets for pancreatic cancer.

Diagnostic performance of microRNA-29a for colorectal cancer: a meta-analysis.

Previous studies have revealed that the expression level of microRNA-29a (miR-29a) was remarkably different in colorectal cancer (CRC) patients and healthy controls, indicating that miR-29a can be used as a diagnostic marker of CRC, but the results have been inconsistent. We conducted this meta-analysis to assess the diagnostic performance of blood-based miR-29a for CRC. We performed a systematic review of studies published over the past two decades to investigate the diagnostic performance of serum miR-29a for the diagnosis of CRC. QUADAS-2 was used to evaluate the quality of the studies. Performance characteristics (diagnostic sensitivity, specificity, and other measures of accuracy) were pooled and examined using random-effect models. Five studies, which included 281 CRC patients and 299 healthy controls, met the inclusion criteria. The summary estimates for miR-29a in CRC diagnoses showed a diagnostic sensitivity of 0.59 (95%CI = 0.53-0.65), a specificity of 0.89 (95%CI = 0.85-0.93), and a diagnostic odds ratio of 12.22 (95%CI = 5.07-29.44). The area under curve and Q value for the summary receiver operating characteristic curves were 0.9128 and 0.8453, respectively. In conclusion, miR-29a may be a novel potential biomarker for CRC diagnosis.

Olfactory mucosal microstructural changes in a rat model of acute rhinosinusitis with dysosmia.

This study aimed to examine olfactory mucosal changes in a rat model of acute rhinosinusitis and to determine how acute rhinosinusitis induces dysosmia. Total 100 rats were divided into the experimental (N = 80) and control (N = 20) groups. An acute rhinosinusitis model was established in the experimental rats. From the experimental group, 20 rats each were sacrificed at 1, 2, 3, or 4 weeks. The control group was sacrificed at 1 week without inoculation. Olfaction was tested by the buried food pellet test before killing the rats, and the duration until the pellet was recorded and analyzed by a statistical software program. Mature olfactory receptor neurons (ORNs) and olfactory ensheathing cells (OECs) were examined. The durations spent by the experimental group in the BFPT were 402.9 ± 9.3, 453.7 ± 7.3, 351.9 ± 8.9, and 278.7 ± 8.1 s at 1, 2, 3, and 4 weeks, respectively. These durations were greater than that of the control group (178.3 ± 6.6 s), and the difference was statistically significant (P < 0.01). A reduction in ORNs and thinning of the epithelium were detected in the experimental group, most notably in the 2nd week and less in the fourth week. OECs were reduced in the 1st week, increased in the 2nd week, and almost completely recovered in the fourth week. In addition, some growing OECs were detected in the experimental group. Acute rhinosinusitis decreased both the mature ORNs and OECs, but OECs increased faster than the ORNs.

Expression of heat shock protein 90 alpha (Hsp90α) in primary neonatal rat myocardial cells exposed to various periods of heat stress in vitro.

The objective of this study was to investigate the mechanism of heat shock protein 90 alpha (Hsp90α) protection against heart damage resulting from heat stress by detecting Hsp90α mRNA, Hsp90α protein, protein localization, and cell damage in primary myocardial cells of neonatal rats in response to heat stress in vitro. The cells were heat-stressed at 42°C in an incubator with 95% air and 5% CO2 for different periods. Levels of Hsp90α, protein localization, enzymes, and cytopathological lesions were detected using Western blot, immunocytochemistry enzymatic assays, and cytopathological techniques. Aspartate aminotransferase, lactate dehydrogenase, and creatine kinase enzyme levels were elevated during heat stress, and acute cellular lesions that were characterized by vacuolar degeneration and necrosis were observed. Hsp90α levels decreased between 10 and 60 min of heat stress and increased after 360 and 480 min, while Hsp90α mRNA decreased after 360 min. These results indicate that heat stress might induce irreversible damage in certain myocardial cells. The elevated Hsp90α level at the end of heat stress and its positive signal in the cytoplasm of myocardial cells after heat stress could be associated with its protective role. Additionally, the consumption of Hsp90α exceeded its production in the first period of treatment.

Quantitative trait loci mapping for kernel row number using chromosome segment substitution lines in maize.

Unveiling the genetic architecture of grain yield and yield-related traits is useful for guiding the genetic improvement of crop plants. Kernel row number (KRN) per ear is an important yield component, which directly affects the grain yield of maize. In this study, we constructed a set of 130 chromosome segment substitution lines (CSSLs), using Nongxi531 as the donor parent and H21 as recipient parent, by continuous backcrossing and selfing. In total, 11 quantitative trait loci (QTL) were detected for KRN by stepwise regression under 3 environmental settings, with 9.87-19.44% phenotypic variation being explained by a single QTL. All 11 QTL were also detected by single-factor ANOVA across the 3 environments tested. Of these 11 QTL, 4 were identified across more than 2 environments, indicating that they are authentically expressed under different environments to control the formation and development of KRN in female maize inflorescences. The CSSLs harbored a greater number of favorable alleles for KRN compared to the H21 line, and could be employed as improved H21 lines in maize breeding programs.

The role of Hsp90α in heat-induced apoptosis and cell damage in primary myocardial cell cultures of neonatal rats.

To understand the mechanism underlying the sudden animal death caused by acute heart failure during heat stress, the relationships among the heat-induced pathological changes and apoptosis and the variations in the levels of protective Hsp90α and its mRNA in the heat-stressed primary myocardial cells of neonatal rats in vitro were studied by cytopathological observation, immunoblotting, RT-PCR, and analysis of the related enzymes. After a period of adaptive cell culture, the myocardial cells were immediately exposed to heat stress at 42°C for 10, 20, 40, 60, 120, 240, 360, and 480 min. Levels of creatine kinase increased from the beginning of heat stress, and the cells exposed to heat stress showed acute cellular lesions characterized by vacuolar degeneration and necrosis after 40 min of heat stress, suggesting that the myocardial cells in vitro were obviously stressed and damaged by higher temperature. The levels of cleaved caspase-3 and cytochrome C, which were related to apoptosis, increased significantly after 40 min of heat stress while the Hsp90α protein level significantly decreased. In contrast, after 6 h of exposure to heat stress, the levels of cleaved caspase-3 and cytochrome C decreased while those of Hsp90α significantly increased, suggesting that early depletion of Hsp90α coincides with a high rate of necrosis and apoptosis in heat-stressed myocardial cells, while the Hsp90α level in surviving cells increases again with significantly less apoptosis after 6 h of heat stress. These findings also indicate that apoptosis of myocardial cells occurs through the activation of the cytochrome C and caspase-3 pathway. The cell repair capacity of Hsp90α is overstrained in the early phase of heat treatment and needs some hours to stabilize. As a result, in the primary myocardial cells in vitro, Hsp90α shows protective activity against damage at the end period of the heat exposure.

Temporal variations of Hsp60 and HSF-1 in primary rat myocardial cells in vitro under heat stress.

The mechanisms involved in sudden animal death due to acute heart failure during heat stress are not well understood. We examined the relationship between heat stress-induced variations of protective Hsp60 and expression of its regulatory factor, HSF-1, in heat-stressed primary myocardial cells of neonatal rats in vitro through cardiac enzyme detection, immunoblotting, immunocytochemistry, and qPCR. Increases in cardiac damage-related enzyme levels demonstrated injury to myocardial cells after heat exposure at 42°C. Hsp60 expression levels fluctuated during heat stress; they decreased significantly after 20 min, then increased at 120 min and decreased again at 360 min after initiation of heat stress. The highest levels of Hsp60 were observed at 240 min, while the lowest were at 60 min. Damage to myocardial cells was characterized by increases in cardiac enzyme levels and low levels of Hsp60 due to functional disorder of myocardial cells at early stages of heat stress. However, the significant induction of hsp60 mRNA levels from the beginning up to 240 min of heat stress was not consistent with the classic regulatory mechanisms that link transcription and translation, suggesting that Hsp60 expression is delayed due to loss of Hsp60 during the early stages of heat stress. hsf-1 mRNA levels were significantly increased from 10 min of heat stress; however, HSF-1 protein levels did not simultaneously increase, indicating that HSF-1 is not the sole regulator of Hsp60 expression.

Hsp110 expression changes in rat primary myocardial cells exposed to heat stress in vitro.

We investigated and described the kinetics of heat shock protein (Hsp) 110 expression and distribution in rat primary myocardial cells exposed to heat stress in vitro. After incubation at 37°C for 72 h, myocardial cells were heat stressed at 42°C for 0, 10, 20, 40, 60, 120, 240, 360, and 480 min. Significant increases in aspartate transaminase, lactate dehydrogenase, and creatine kinase enzymatic activities in the myocardial cell culture media were observed during heat stress, suggesting that the integrity of the myocardial cells was altered. Immunocytochemical analysis revealed that the expressed Hsp110 was constitutively localized in the cytoplasm and in the nuclei in small amounts characterized by a granular pattern. Nuclear Hsp110 levels increased significantly after 240 min of heat stress compared with levels in the control. The overall levels of Hsp110 expression increased significantly after 20 min. After 240 min, Hsp110 levels were approximately 1.2-fold higher than those in the control. Increasing levels of hsp110 messenger RNA detected using real-time quantitative polymerase chain reaction were observed after 20 min of heat stress, and the levels peaked with a 10-fold increase after 240 min of heat stress. These results indicate that the expression of Hsp110 in primary myocardial cells in vitro is sensitive to hyperthermic stress and that Hsp110 is involved in the potential acquisition of thermotolerance after heat stress. Therefore, Hsp110 might play a fundamental role in opposing and alleviating heat-induced damage caused by hyperthermic stress in primary myocardial cells.