Chemical structures, biosynthesis, bioactivities, and utilisation values for the diterpenes produced in tobacco trichomes.

Cembranoids and labdanes are two important types of diterpenes in tobacco (Nicotiana genus) that are predominantly found in the leaf and flower glandular trichome secretions. This is the first systematic review of the biosynthesis, chemical structures, bioactivities, and utilisation values of cembranoid and labdane diterpenes in tobacco. A total of 131 natural cembranoid diterpenes have been reported in tobacco since 1962; these were summarised and classified according to their chemical structure characteristics as isopropyl cembranoids (1-88), seco-cembranoids (89-103), chain cembranoids (104-123), and polycyclic cembranoids (124-131). Forty natural labdane diterpenes reported since 1961 were also summarised and divided into epoxy side chain labdanes (132-150) and epoxy-free side chain labdanes (151-171). Tobacco cembranoid and labdane diterpenes are both formed via the methylerythritol 4-phosphate pathway and are synthesised from geranylgeranyl diphosphate. Their biosynthetic pathways and the four key enzymes (cembratrienol synthase, cytochrome P450 hydroxylase, copalyl diphosphate synthase, and Z-abienol cyclase) that affect their biosynthesis have been described in detail. A systematic summary of the bioactivity and utilisation values of the cembranoid and labdane diterpenes is also provided. The agricultural bioactivities associated with cembranoid and labdane diterpenes include antimicrobial and insecticidal activities as well as induced resistance, while the medical bioactivities include cytotoxic and neuroprotective activities. Further research into the cembranoid and labdane diterpenes will help to promote their development and utilisation as plant-derived pesticides and medicines.

[Management Practice of Outsourcing Hospital Tests to Independent Clinical Laboratories].

With the development of medical technology and the deepening of medical reform, hospital laboratory test continues to expand. Affected by factors such as technology and cost, the business of outsourcing laboratory test to independent clinical laboratories develops rapidly. However, this cooperation mode has not been carried out for a long time and lacks systematic management experience. Through the analysis of the motivation of hospital delivery, this study expounds the classification, judgment basis and requirements for suppliers of third-party clinical laboratory delivery, as well as the operation practice of laboratory test delivery, so as to provide reference for more standardized and effective testing delivery for hospitals.

Effect of Lanthanum Addition on Formation Behaviors of Inclusions in Q355B Weathering Steel.

The effect of lanthanum addition on the formation behaviors of inclusions in Q355B weathering steel was investigated by laboratory experiments and thermodynamic calculations. The results demonstrate that the main inclusions in weathering steel without La addition are large-sized irregular Al2O3 and MnS, with an average size of about 5.35 µm. As La content increases from 0.0075 to 0.0184 wt.%, the dominant inclusions transform from MnS, LaAlO3, and Al2O3-LaAlO3 into MnS, La2O3, and LaAlO3-La2O3. Meanwhile, the average size of inclusions significantly decreases from 3.4 to 2.48 µm and the distribution is more dispersive. When the La content increases to 0.0425 wt.%, the original MnS and Al2O3 inclusions are completely modified into La2O2S and La2O3 but the inclusions demonstrate serious agglomeration and growth. The thermodynamic calculations indicate that Al2O3 and various lanthanum-containing inclusions are formed in the liquid phase. As the La content in molten steel increases from 0 to 0.0425 wt.%, the Al2O3 inclusion is inclined to be modified into lanthanum oxide and lanthanum oxysulfide and the modification process is Al2O3 → LaAlO3 → La2O3 → La2O2S, which is very consistent with the experimental observations.

Effect of Cold Rolling and Annealing on the Microstructure and Texture of Erbium Metal.

Erbium metal with purity ≥ 99% was cold rolled to 30%, 40%, 50%, and 60% deformations and the Er metal of 60% deformation was annealed at different temperatures for 1 h. The effect of cold rolling deformation and annealing on the microstructure and texture evolution of Er metal was investigated by XRD, EBSD, Microhardness tester, and OM. P is the orientation index, which is used to judge the preferred orientation. The research results showed that grains were broken and refined gradually with increasing deformation, the average grain size was 3.37 µm, and the orientation distribution was uniform for 60% deformation; deformation twins appeared in the grain when the deformation was less than 40%, which contributed to the generation of (0001) plane orientation. Comparing with the initial state, the (011-0) plane orientation gradually weakened and the (111-0) plane orientation had a trend of further strengthening with the increasing deformation; the (1-21-0) plane orientation remained unchanged, but there was a gradual weakening trend when the deformation was greater than 50%. For 60% deformation of Er metal, the deformed microstructure was replaced by fine equiaxed grains with the increasing annealing temperature, and the high-performance Er metal with fine and uniform equiaxed grains can be obtained under annealing at 740 °C for 1 h.

Effects of dietary monocalcium phosphate supplementation on the anti-oxidative capacity, anti-bacteria function in immune organs of obscure puffer (Takifugu obscurus) after infection with Aeromonas hydrophila.

The purpose of the present study was to explore the impaired anti-bacteria ability in immune organs and immune systems of obscure puffer induced by chronic dietary phosphorus (P) deficiency. Fish were fed diets supplemented with 6 g/kg P (P6) and 0 g/kg P (P0) respectively for 15 weeks, and lower final body weight, feed intake, weight gain, whole body P content and bone P content were observed in fish fed P0 diet (P < 0.05). Then the fish were continued to feed for 3 weeks and intraperitoneal injection with PBS (P6+PBS) and Aeromonas hydrophila (A.hydrophila) (P6 + A.hydrophila and P0 + A.hydrophila), and sampled at 3, 6, 12 and 24 h. The results showed that dietary P deficiency lowered survival rate, total hemocyte count, whereas enhanced ROS production and apoptosis rate of obscure puffer compared to the 6 g/kg P supplemented group after infection. Moreover, compared to the P sufficient group, puffer fish fed P deficient diet decreased the expressions of antioxidant genes catalase (cat) and glutathione reductase (gr), immune-related genes toll-like receptor 2 (tlr-2) and anti-inflammatory factors transforming growth factor ß1 (tgf-ß1) and interleukin 11 (il-11) while increased pro-inflammatory cytokines tumor necrosis factor α (tnf-α), interleukin 1ß (il-1ß) and interleukin 8 (il-8) in head kidney post-infection. In addition, dietary P deficiency decreased the hepatic gene expressions of anti-apoptotic factor B-cell lymphoma 2 (bcl-2) and bax-inhibitor 1 (bi-1), accompanied by increasing the mRNA expressions of pro-apoptotic factor caspase 3, caspase 8 and caspase 9 compared to the P sufficient group after A.hydrophila infection. In conclusion, dietary P deficiency impaired the anti-bacteria function of the immune system as well as immune organs by increasing oxidative stress and aggravating the inflammatory response and apoptosis in obscure puffer under the A.hydrophila challenge.

Effects of dietary Senecio scandens buch-ham extracts on growth performance, plasma biochemical, histology and the expression of immune-related genes in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀).

The study mainly aimed at the effects of dietary Senecio scandens buch-ham extract (SSBE) on the growth performance, body composition, plasma biochemical index, intestinal and liver histology and the expression of antioxidant, apoptosis and inflammatory related genes in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀). Basal diets supplemented with SSBE (10:1) 0%, 0.05%, 0.1%, 0.2% and 0.4% were fed hybrid grouper for 8 weeks. The results showed that WGR and SGR were significantly increased in the week 2 and week 4 in Diet 0.05% group (P < 0.05). The total protein, globulin and albumin significantly increased whereas alanine aminotransferase, triglyceride and alkaline phosphate in the plasma were significantly decreased in Diet 0.1% group (P < 0.05). The villi length, width, muscle thickness and the cross-sectional area of intestine were improved in Diet 0.05% and Diet 0.1% group. The expression levels of PPAR-α and CPT-1 in the liver of hybrid grouper were significantly increased following the supplementation of SSBE (P < 0.05). The expression levels of antioxidant related genes (CAT, GPX, GR and Keap1) and anti-inflammatory factor (IL-10) in liver, head kidney and spleen of hybrid grouper decreased significantly (P < 0.05). In addition, diets supplemented with 0.05%-0.1% SSBE had a good liver-protecting effect, but it would have a detrimental effect on hepatocytes when the content exceeds 0.2%. The above results indicated that the suitable additive amount of SSBE in hybrid grouper feed was 0.05%-0.1%.

Xiaochaihu Decoction reduces hepatic steatosis and improves D-GalN/LPS-induced liver injury in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀).

Excessive lipid accumulation and chemical abuse can induce fatty liver diseases in fish, but the underlying mechanism and therapies are unknown. The present study aims to evaluate the effects of Xiaochaihu Decoction (XCHD) on the growth performance, lipid metabolism and antioxidant function of hybrid grouper in vitro and in vivo, and provide evidence as to whether it can be potentially used as a medicine for liver diseases in aquaculture. In vitro, steatosis model of hybrid grouper primary hepatocytes were incubated for 48 h in control or lipid emulsion (LE)-containing medium with or without 24 h post-treatment with XCHD. XCHD treatment reversed the LE-induced intracellular lipid accumulation, cell viability and hepatocytes morphological structure. In vivo, a total of 300 hybrid grouper with an average initial weight of 25.43 ±â€¯0.18 g were fed diets containing five graded levels of XCHD at 150-1200 mg/kg diet for 8 weeks. After that, a challenge trial was conducted by injection of D-GalN/LPS to induce liver injury. As a result, dietary supplementation with 150-300 mg/kg XCHD diets can significant improve growth performance and feed utilization (P < 0.05). Dietary XCHD down-regulated the expression of lipogenic-related genes (G6PD, DGAT2 and ME1) and up-regulated lipolysis-related genes (ATGL, PPARα and LPL) expression in the liver of hybrid grouper. Livers challenged with D-GalN/LPS exhibited extensive areas of vacuolization with the disappearance of nuclei and the loss of hepatic architecture. These pathological alterations were ameliorated by XCHD treatment. XCHD significantly down-regulated the D-GalN/LPS induced apoptosis-related genes caspase-3, caspase-9 and p53 mRNA expression and up-regulated the antioxidant-related genes CAT and MnSOD mRNA expression in dose dependent manner, respectively. XCHD potently reduced hepatic lipid accumulation and enhanced antioxidant capability in hybrid grouper and may be a potential fish-feed additive to prevent fatty liver diseases onset and progression.

Dietary Radix Bupleuri extracts improves hepatic lipid accumulation and immune response of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀).

In this study, two experiments were performed to explore the effect of Radix Bupleuri extracts (RBE) on growth, lipid deposition and metabolism and immune response of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) using in vitro and in vivo models. In vitro, we used 2 ml/L 20% lipid emulsion (LE)-induced steatosis in hybrid grouper primary hepatocytes, then RBE (200, 400 and 800 µg/ml) was added to the hepatocytes after (post-treatment) the incubation with 20% LE (2 ml/L) in the culture medium. We found that RBE markedly increased cell viability, which were consistent with hepatocytes morphological structure examination and lipid metabolism and immune related genes study. The above result suggested that RBE has a protective effect on this model of hepatocytes damage. In vivo, five graded levels of RBE at 0, 200, 400, 800 and 1600 mg/kg diet were supplemented to a basal diet with 15% lipid levels (high lipid), and fed to a total of 300 hybrid grouper with an average initial weight of 25.58 ±â€¯0.05 g for 8 weeks. Growth performance, liver histology, plasma biochemical parameters, and expression of genes involved in lipid metabolism and immune-related were measured. The study indicated that dietary RBE significantly improved growth performance and feed utilization and reduced hepatosomatic index. Dietary supplementation with 200-800 mg/kg RBE diets effectively decreased serum ALP, ALT, AST and LDH contents in fish. Furthermore, adipogenesis relative mRNA levels of DGAT2, G6PD, ME1 and DGKα in fish fed 200-400 mg/kg RBE diets were lower (P < 0.05) than in those fed RBE0 diets, while dietary supplementation with 200-800 mg/kg RBE diets up-regulated lipolysis-related genes (CPT1, LPL and PPARα) expression in the liver of hybrid grouper. Moreover, dietary RBE down-regulated the expression of apoptosis-related genes (caspase-9), up-regulated the expression of antioxidant genes (CAT) and immune-related genes (MHC2, IKKα and TGF-ß1). Thus, our data suggest that RBE suppressed lipid accumulation and enhanced immune capability in hybrid grouper both in vitro and in vivo. These results offer new insight into RBE as a hepatoprotective in fish.

Effects of dietary dandelion extracts on growth performance, liver histology, immune-related gene expression and CCl4 resistance of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀).

The study investigated the effects of dietary supplementation with dandelion extracts (DE) on growth performance, feed utilization, body composition, serum biochemical, liver histology, immune-related gene expression and CCl4 resistance of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀). A basal diet supplemented with DE at 0% (diet 0%), 0.1% (diet 0.1%), 0.2% (diet 0.2%), 0.4% (diet 0.4%) and 0.8% (diet 0.8%) were fed to hybrid grouper for 8 weeks. The results revealed that dietary DE had not a significant impact on growth performance and feed utilization (P > 0.05), but it could decrease the percent of crude lipids in whole body and increase the percent of crude protein in muscle (P < 0.05). Dietary DE increased the mRNA levels of antioxidant enzymes (catalase, glutathione peroxidase and glutathione reductase) and reduced inflammatory factor in the spleen and head-kidney of fish (P < 0.05), but reduced the expression of the liver antioxidant gene except for glutathione reductase (P < 0.05). Dietary supplementation with 0.2%-0.4% DE could effectively improve liver health. After injection of CCL4 by 72 h, fish fed Diet0.2% and Diet0.4% showed regular hepatocyte morphology while fish fed Diet 0%, Diet 0.1% and Diet 0.8% showed hepatocyte damage. Higher survival rate and total blood cell count was observed in fish fed 0.1%-0.4% dietary DE (P < 0.05). In conclusion, DE could be used as a functional feed additive to enhance liver function of farmed fish. The best level of it should be between 0.2% and 0.4%.

Liver transcriptome analysis and de novo annotation of the orange-spotted groupers (Epinephelus coioides) under cold stress.

Cold stress has caused great economic loss in fish culture worldwide. Orange-spotted grouper (Epinephelus coioides) is one of the most serious lost aquatic animals in 2016 cold fronts in South China. However, the molecular mechanism of grouper's cold resistance has remained largely unknown. In the present study, HiSeq™2000 (Illumina) was used to analyze the transcriptomic profiles of the liver from grouper under control temperature (CT, 28 °C) and low temperature (LT, 13 °C). Two normalized liver cDNA libraries of CT and LT groups were created. We obtained 51,944,970 and 51,905,036 clean reads from CT and LT groups, respectively. Comparing the LT group to the CT group, a total of 5905 significantly differentially expressed genes (DEGs) were identified, including 2093 up-regulated unigenes and 3812 down-regulated unigenes. GO annotation and functional enrichment analysis indicated that all of the DEGs were classified into three categories: biological process (23 subclasses), cellular component (18 subclasses) and molecular function (13 subclasses). KEGG analysis of the DEGs showed that 2732 DEGs were annotated to 253 signaling pathways. The most highly enriched pathways were cell adhesion molecules, Staphylococcus aureus infection, PPAR signaling pathway, Vibrio cholerae infection, primary immunodeficiency, fatty acid elongation, and we found cold stress mainly affects immunity, metabolic and signal transduction. Thirteen of the DEGs were further validated by qRT-PCR. Our results provide valuable information for further analysis of the mechanisms of groupers response under cold stress.

Effects of replacing fish meal with soybean meal on growth performance, feed utilization and physiological status of juvenile obscure puffer, Takifugu obscurus.

An 8-week feeding trial was conducted to evaluate the growth performance, feed utilization and physiological status of obscure puffer, Takifugu obscurus (13.03 ±â€¯0.14 g) fed diets in which fish meal (FM) was replaced with various levels of dehulled and defatted soybean meal (SBM): 0% (SBM0), 15% (SBM15), 30% (SBM30), 45% (SBM45), 60% (SBM60) and 75% (SBM75). No significant differences were observed in weight gain and specific growth rate (SGR) of fish when FM replacement level was lower than 30%, and the broken-line model of SGR showed the maximum replacement level was 40%. Fish fed the SBM-containing diets had a lower red blood cell value compared to the control. The hemoglobin and methemoglobin values showed a declining tendency as dietary SBM level increased. Plasma triacylglycerol, cholesterol and low-density lipoprotein cholesterol levels also showed a decreasing trend that was associated with the reduced crude lipid content of whole body as dietary SBM level increased. The activities of alanine aminotransferase and aspartate aminotransferase in fish fed the SBM-containing diets were all higher than those fed the control diet while glutathione peroxidase and catalase activities were lower than the control group. Results indicated that up to 40% FM protein, based on the broken-line analysis of SGR, can be replaced with SBM in diet for obscure puffer juveniles with supplemental lysine, methionine and taurine.

The hepatoprotective effects of Radix Bupleuri extracts against D-galactosamine/lipopolysaccharide induced liver injury in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀).

The present study is aiming at evaluating the hepatoprotective of Radix Bupleuri extracts (RBE) on the d-galactosamine/lipopolysaccharide (D-GalN/LPS) induced liver injury of hybrid grouper in vitro and in vivo. In vitro, RBE (0, 200, 400 and 800 µg/ml) was added to the hybrid grouper primary hepatocytes before (pretreatment) the incubation of the hepatocytes with D-GalN (20 mM) plus LPS (1 µg/ml) in the culture medium. RBE at concentrations of 200, 400 and 800 µg/ml significantly improved cell viability and inhibited the elevation of TNF-α, IL-1ß and IL-6 and significantly down-regulated the caspase-3, caspase-9 and P53 mRNA levels. In vivo administration of RBE at the doses of 0, 200, 400, 800 and 1600 mg/kg in the diet for 8 weeks prior to D-GalN (500 mg/kg) and LPS (20 µg/kg) intoxication. The study indicated that the RBE not only ameliorated liver injury, as evidenced by well-preserved liver architecture, but also significantly increased hepatic antioxidant enzymes activities in the D-GalN/LPS-induced liver injury animal model. Further demonstrating the protective effects of the RBE, we found that pretreatment with the RBE up-regulated the expression of antioxidant genes (GPx and MnSOD), while down-regulated apoptosis-related genes (caspase-3, caspase-9 and P53), immune related genes (MHC2 and TLR3) and pro-inflammatory cytokines (TOR and IKKα) mRNA expression in the liver of hybrid grouper. In brief, the present study showed that RBE can protect hepatocyte injury induced by D-GalN/LPS through elevating antioxidant enzyme activity and suppressing apoptosis and immune inflammatory responses. The results support the use of RBE as a hepatoprotective in fish.

Astragalin and dihydromyricetin as adjuncts to histidine­tryptophan­ketoglutarate cardioplegia enhances protection during cardioplegic arrest.

The present study used an in vitro model of cold cardioplegia in isolated working rat hearts to evaluate the possible effects of two flavonoids, astragalin and dihydromyricetin, as adjuncts to histidine­tryptophan­ketoglutarate (HTK) cardioplegia. The following three groups of male Sprague Dawley rats were evaluated: The HTK group, treated with HTK alone; the HTK­A group, treated with 10 µmol/l astragalin; and the HTK­D group, treated with 10 µmol/l dihydromyricetin. Isolated rat hearts were perfused with Krebs­Henseleit buffer for 30 min and incubated with the respective cardioplegic solution for 6 h at 4˚C. Subsequently, astragalin or dihydromyricetin was added to the cardioplegic solutions. Following 30 min of reperfusion, the left ventricular developed pressure (LVDP), maximum up/down rate of left ventricular pressure (±dp/dtmax) and heart rate were documented as indices of myocardial function using a physiological recorder. Myocardial infarct size (IS) was estimated using 2,3,5­triphenyltetrazolium chloride staining. Lactate dehydrogenase (LDH) and creatine kinase (CK) levels were also determined to assess the degree of cardiac injury. Cardiomyocyte apoptosis analysis was performed using an in situ cell death detection kit. In addition, malondialdehyde (MDA), superoxide dismutase (SOD), interleukin­6 (IL­6), tumor necrosis factor­α (TNF­α), C­reactive protein (CRP) levels, as well as the glutathione/glutathione disulfide (GSH/GSSG) ratio were determined and analyzed using ELISA kits. The protein levels of caspase­9 and B­cell lymphoma­2 (Bcl­2) were determined using western blot analysis. The results demonstrated that exposure to astragalin or dihydromyricetin significantly improved the recovery of LVDP (P<0.05 and P<0.01, respectively), the +dP/dtmax (P<0.05 for dihydromyricetin only) and the ­dP/dtmax (P<0.05 and P<0.01, respectively), increased SOD levels (P<0.05 and P<0.01, respectively) and GSH/GSSG ratios (P<0.05), reduced myocardial IS (P<0.05 and P<0.01, respectively), decreased CK, LDH, IL­6 (all P<0.05 and P<0.01, respectively), MDA (P<0.05), CRP (P<0.05) and TNF­α levels (P<0.05 and P<0.01, respectively), increased Bcl­2 levels (P<0.01) and decreased caspase­9 levels (P<0.01). The results indicated that the addition of either flavonoid (particularly dihydromyricetin) to HTK enhances protection during ischemia, decreases myocardial dysfunction by enhancing anti­inflammatory activities, attenuates myocardial oxidative injury and prevents apoptosis during ischemia/reperfusion.

Human Papillomavirus Infection and Vaccination: Awareness and Knowledge of HPV and Acceptability of HPV Vaccine among Mothers of Teenage Daughters in Weihai, Shandong, China.

In preparation for the introduction of human papillomavirus (HPV) vaccine, we investigated awareness and knowledge of HPV/HPV vaccine and potential acceptability to HPV vaccine among mothers with a teenage daughter in Weihai, Shandong, China. A cross-sectional survey was conducted in 2013 with a sample of 1850 mothers who had a daughter (aged 9-17 years) attending primary, junior and senior high schools. In the final sample (N = 1578, response rate 85.30%), awareness of HPV was reported by 305 (19.32%) mothers. Awareness varied significantly by daughter's age (P<0.01), mother's education level (P<0.01), mother's occupation (P<0.01), household income (P<0.01) and residence type (P<0.01). Knowledge about HPV/HPV vaccine was poor with a mean total score of 3.56 (SD = 2.40) out of a possible score of 13. Mothers with a higher education level reported higher levels of knowledge (P = 0.02). Slightly more than one-fourth (26.49%) of mothers expressed their potential acceptability of HPV vaccine for their daughters. Acceptability increased along with increased daughters' age (P<0.01), household income (P<0.01) and knowledge level (P<0.01). House wives and unemployed mothers had the highest acceptability (P<0.01). The most common reasons for not accepting HPV vaccination were "My daughter is too young to have risk of cervical cancer (30.95%)", "The vaccine has not been widely used, and the decision will be made after it is widely used (24.91%)", "Worry about the safety of the vaccine (22.85%)". Awareness and knowledge of HPV/HPV vaccines are poor and HPV vaccine acceptability is low among these Chinese mothers. These results may help inform appropriate health education programs in this population.

Molecular characterization of coxsackievirus A21 in Shandong, China.

Coxsackievirus A21 (CV-A21) is a rarely detected serotype belonging to the species Enterovirus C (EV-C). In this study, we report the isolation and genetic characterization of CV-A21 in Shandong Province, China, during 1997 to 2013. A total of 13 strains were obtained from surveillance of cases of acute flaccid paralysis (AFP) (n = 9) and from environmental sewage (n = 4). Sequence comparison of the VP1 genes revealed high nucleotide sequence similarity (94.1 % to 99.8 % identity) among these Shandong strains during the period of 17 years and 75.8 % to 98.5 % sequence identity to foreign strains. Bayesian phylodynamic evolutionary analysis of Shandong and global CV-A21 VP1 sequences revealed that the inferred CV-A21 ancestral sequence dated back to 1750 (1643-1841) and evolved with 2.943 × 10(-3) substitutions per site per year. Alignment of the deduced VP1 amino acid sequences revealed changes that might alter the hydropathicity of the encoded protein. The complete genome of one strain from 2013 was sequenced and evidence of recombination was detected by similarity plot and bootscanning analyses. This study describes the complete genome characterization and molecular epidemiology of CV-A21 in China and gives further insight into CV-A21 evolution.

Environmental Surveillance of Genogroup I and II Noroviruses in Shandong Province, China in 2013.

Noroviruses are the most common cause of epidemic gastroenteritis. However, the case-based surveillance is limited in China. In this study, we analyzed the results of environmental surveillance conducted in two cities of Shandong Province, China from January to December in 2013. Twenty-four sewage samples were collected and concentrated via membrane absorption/elution method. After reverse transcription-PCR, cloning and sequencing on ORF2 region, norovirus nucleic acid was detected in all 24 sewage samples. A total of 403 norovirus sequences of 16 genotypes were detected, among which GII.3 (22.6%), GI.2 (17.1%), GI.5 (13.4%), GI.3 (11.9%), GII.4 (7.7%), and GII.6 (6.7%) were the 6 most common genotypes. Phylogenetic analysis revealed multiple lineages within most common genotypes, especially in GI.3, whereas all GII.4 sequences belonged to Sydney 2012 strain. Recombination events were observed in 5 GI and 4 GII sequences within or near the ORF1/ORF2 overlap. This is the first report on systematic environmental surveillance on norovirus in China. The data presented here reveal co-circulation and high genetic diversity of multiple norovirus genotypes in the two cities, and suggest continued environmental surveillance can provide valuable information on norovirus circulation in the population.

The Influence of Casing-Sand Adhesion on Cementing Bond Strength.

In the petroleum industry, one of the most serious problems encountered during cementing is the failure at the bonding interface. Many measures including casing-sand adhesion have been developed to improve cementing bond strength. However, due to the lack of detailed study of the technique, many questions remain. The primary goal of this study is to investigate the influence of casing-sand adhesion on cementing bond strength, and to optimize parameters. An orthogonal experiment and a supplementary experiment were conducted. The results indicated that casing-sand adhesion can improve the cementing bond strength. The priority orders of key factors are: sand grain size, sand coverage, adhesive curing temperature and adhesive curing time. The optimal parameters recommended for application are: 1.6mm~1.9mm sand grain size, 60%~70% sand coverage, 30°C curing temperature and 60 hours curing time.