ABSTRACT Expression of housekeeping genes is relatively constant in different tissues and cells by RT-qPCR analysis. Housekeeping genes (HGs) are usually utilized as the reference to evaluate and compare mRNA expression abundances of target genes in different cells or tissues sampled. However, the expression stabilities of different HGs in diverse samples may appear divergence. Currently, there is no exact reference data of HGs in hen ovarian follicular tissues during egg-laying period available yet. In this study, we detected the expression of 18SrRNA, ACTB, HOXC8, GAPDH, alpha-A, and alpha-D mRNA in the varied-size ovarian follicles (1-8 mm in diameter and F5), hearts, livers, spleens, lungs, and breast muscles of the laying hens by RT-qPCR, to analyze the results via Ct value, geNorm, Normfinder, and Bestkeeper. The data showed that the expression levels of 18SrRNA, alpha-A, and alpha-D transcripts were more significantly stable than the other three genes for normalizing mRNA expression in the hen ovarian follicles examination. Moreover, alpha-D, 18SrRNA, and alpha-A were also most suitable for the expression normalization in the tissues of the heart, liver, spleen, lung and breast muscle. In contrast, 18SrRNA has the most stable mRNA expression levels in all tissues sampled, so it can serve as an excellent inner control for the evaluation of the transcription levels in chickens. It is a remarkable fact that HOXC8 as a candidate reference should be avoided. Our study establishes a set of stably expressed candidate inner references in the hen ovarian follicles and several tissues, it firstly provided an exact data for validation of the inner references in normalizing transcription levels of a target gene in chickens.

The stabilization of roadways in soft rock under high stress has always been a major concern for deep underground coal mines. This paper describes a case study focusing on the stability of a soft-rock roadway in Yuandian coal mine in Anhui Province, China. The Beiyi main return roadway in this mine was primarily supported by U-steel support; however, considerable deformation and severe failures occurred. Therefore, a new support design, the yielding bolt - grouting support design, was proposed based on the physical and mechanical parameters, in situ stress measurements, and the failure characteristics of the roadway. The mechanics of the yielding bolt - grouting support was analysed and compared to the normal bolt - grouting support. A field experiment was conducted in a 100 m long section of the roadway. The monitoring results showed that compared with the old support, the new support design reduced the deformations of roof, floor, and rib by 41.4%, 56.1%, and 64.7% respectively, and the maximum deformation rate decreased from 15.12 mm/d to 9.28 mm/d. This case study indicates that the yielding bolt - grouting support design is an effective method to support roadways in soft rock under high stress.

Immune thrombocytopenia (ITP) is a disease characterized by isolated thrombocytopenia. Abnormal effector T cell activation is an important mechanism in the pathogenesis of ITP. Regulatory T cells (Treg) have a strong immunosuppressive function for T cell activation and their importance in the pathophysiology and clinical treatment of ITP has been confirmed. Myeloid-derived suppressor cells (MDSCs) are other immunosuppressive cells, which can also suppress T cell activation by secreting arginase, iNOS and ROS, and are essential for Treg cells’ differentiation and maturation. Therefore, we speculate that MDSCs might also be involved in the immune-dysregulation mechanism of ITP. In this study, we tested MDSCs and Treg cells in peripheral blood samples of twenty-five ITP patients and ten healthy donors. We found that MDSCs and Treg cells decreased simultaneously in active ITP patients. Relapsed ITP patients showed lower MDSCs levels compared with new patients. All patients received immunosuppressive treatment including dexamethasone alone or in combination with intravenous immune globulin. We found that MDSCs’ level after treatment correlated with platelet recovery. Our study is the first that focused on MDSCs’ role in ITP. Based on our results, we concluded that circulating MDSCs could predict disease activity and treatment response in ITP patients. This preliminary conclusion indicates a substantial significance of MDSCs in the pathophysiology and clinical treatment of ITP, which deserves further investigation.

The effects of ultra-high pressure (UHP) treatment on the chemical properties, colour and sensory quality of young red wine were studied. UHP did not significantly affect the alcohol content, and the methanol content was higher than that of the control, with the latter first increasing and then decreasing with the increase in the pressure or period of treatment. The glycerol content was also markedly changed by UHP, although without a regular pattern. The pH value was not markedly changed by the treatments, and the contents of total acids and volatile acids were scarcely affected. The fructose and glucose contents were clearly changed according to the different treatment conditions. The tartaric acid, citric acid and lactic acid levels showed evident changes, whereas the malic acid level was not changed by UHP treatment. The trend of these changes was similar to what occurs during natural ageing. As the pressure of the treatment was increased to 200 MPa, the chroma value increased, followed by a decline upon higher pressure treatments. The hue of the wine was significantly changed by treatment at different pressures, reaching the highest value after treatment at 400 MPa and then decreasing with increasing pressure. The chroma and hue values of the wine were changed significantly according to the duration of the UHP. After UHP, the appearance, aroma and taste of the wine was improved, with the score for appearance obviously increased. When the wine was treated at 500 MPa for 30 min, its sensory quality received the highest score.

The Helanshan Donglu wine zone (China) is one of the most successful wine region of China, and the phenolic characterisation and antioxidant capacity of the primarily young, monovarietal wines from there were evaluated. The result showed that Helanshan Donglu wines contained abundant phenolic compounds, especially flavan-3-ols, and possessed high antioxidant capacity. In different grape varieties, the phenolic profiles varied greatly in both red and white wines. For red wines, Cabernet Sauvignon represented the highest total phenols (2 631 mg/L GAE), total flavonoids (1 840.83 mg/L CTE) and antioxidant capacity. Gamay and Cabernet Gernischt possessed the highest total flavan-3-ols (1 108.08 mg/L CTE) and total anthocyanins (258.78 mg/L CGE). Amongst the white wines, Chardonnay and Chenin blanc showed the highest phenolic contents and antioxidant capacity. (+)-Catechin and (-)-epicatechin were dominant phenolic constituents in both the red and white wines. Gallic acid and salicylic acid were the second most abundant in red wines, while gentisic acid was the second most abundant in white wines.

Gut-derived endotoxin and pathogenic bacteria have been proposed as important causative factors of morbidity and death during heat stroke. However, it is still unclear what kind of damage is induced by heat stress. In this study, the rat intestinal epithelial cell line (IEC-6) was treated with heat stress or a combination of heat stress and lipopolysaccharide (LPS). In addition, propofol, which plays an important role in anti-inflammation and organ protection, was applied to study its effects on cellular viability and apoptosis. Heat stress, LPS, or heat stress combined with LPS stimulation can all cause intestinal epithelial cell damage, including early apoptosis and subsequent necrosis. However, propofol can alleviate injuries caused by heat stress, LPS, or the combination of heat stress and LPS. Interestingly, propofol can only mitigate LPS-induced intestinal epithelial cell apoptosis, and has no protective role in heat-stress-induced apoptosis. This study developed a model that can mimic the intestinal heat stress environment. It demonstrates the effects on intestinal epithelial cell damage, and indicated that propofol could be used as a therapeutic drug for the treatment of heat-stress-induced intestinal injuries.

Recharge estimation in arid and semi-arid areas is very challenging. The chloride mass balance method applied in western South Africa fails to provide reliable recharge estimates near coastal areas. A relationship between rainfall events and water level fluctuations (WLF) on a monthly basis was proposed in the rainfall infiltration breakthrough (RIB) model for the purpose of groundwater recharge estimation. In this paper, the physical meaning of parameters in the CRD and previous RIB models is clarified, and the RIB model is reviewed with the algorithm improved to accommodate various time scales, namely, daily, monthly and annual scales. Recharge estimates on a daily and monthly basis using the revised RIB approach in 2 study areas, one in a sandy alluvial aquifer (Riverlands) and the other in the Table Mountain Group (TMG) shallow unconfined aquifer (Oudebosch), are presented, followed by sensitivity analysis. Correlation analysis between rainfall and observed WLF data at daily scale and monthly scale, together with recharge estimates obtained from other methods, demonstrates that the RIB results using monthly data are more realistic than those for daily data, when using long time series. Scenarios using the data from Oudebosch with different rainfall and groundwater abstraction inputs are simulated to explore individual effects on water levels as well as recharge rate estimated on a daily basis. The sensitivity analysis showed that the recharge rate by the RIB model is specifically sensitive to the parameter of specific yield; therefore, the accurate representative specific yield of the aquifer needs to be selected with caution. The RIB model demonstrated in these two cases can be used to estimate groundwater recharge with sufficiently long time series of groundwater level and rainfall available in similar regions. In summary, the RIB model is best suited for shallow unconfined aquifers with relatively lower transmissiv-ity; the utility of the RIB model for application in different climatic areas under different hydrogeological conditions needs to be further explored.

Massive hepatectomy associated with infection induces liver dysfunction, or even multiple organ failure and death. Glycyrrhizin has been shown to exhibit anti-oxidant and anti-inflammatory activities. The aim of the present study was to investigate whether glycyrrhizin could attenuate endotoxin-induced acute liver injury after partial hepatectomy. Male Wistar rats (6 to 8 weeks old, weighing 200-250 g) were randomly assigned to three groups of 24 rats each: sham, saline and glycyrrhizin. Rats were injected intravenously with lipopolysaccharide (LPS) 24 h after 70% hepatectomy. Glycyrrhizin, pre-administered three times with 24 h intervals 48 h before hepatectomy, prolonged the survival of rats submitted to partial hepatectomy and LPS injection, compared with saline controls. Glycyrrhizin was shown to attenuate histological hepatic changes and significantly reduced serum levels of aspartate aminotransferase, alanine aminotransferase, and lactic dehydrogenase, at all the indicated times (6 rats from each were sacrificed 1, 3, 6, and 9 h after LPS injection), compared with saline controls. Glycyrrhizin also significantly inhibited hepatocyte apoptosis by down-regulating the expression of caspase-3 and inhibiting the release of cytochrome C from mitochondria into the cytoplasm. The anti-inflammatory activity of glycyrrhizin may rely on the inhibition of release of tumor necrosis factor-a, myeloperoxidase activity, and translocation of nuclear factor-kappa B into the nuclei. Glycyrrhizin also up-regulated the expression of proliferating cell nuclear antigen, implying that it might be able to promote regeneration of livers harmed by LPS. In summary, glycyrrhizin may represent a potent drug protecting the liver against endotoxin-induced injury, especially after massive hepatectomy.