Simulation of FBR-Fenton/GAC process for recalcitrant industrial wastewater treatment with a computational fluid dynamics-kinetic model framework.

An integrated computational fluid dynamics (CFD)-kinetic model framework was developed to numerically describe the hydrodynamic and kinetic phenomena in a liquid-solid two phases Fluidized-bed reactor Fenton/granular activated carbon (FBR-Fenton/GAC) system. The model obtained excellent accuracy for predicting chemical oxygen demand (COD) removal in reverse osmosis concentrate (ROC) treatment under different operation conditions. Hydrodynamic evaluation demonstrated that under the quasi-steady state, the GAC particles were uniformly circulated in the bed region with two pairs of counter-rotating recirculation cells, and a clear interface layer formed between the solid and the liquid phases. Superficial liquid velocity highly affected the fluidized bed expansion and solid volume fraction, while its impact on the overall COD removal efficiency was negligible. Chemical evaluation revealed that GAC/H2O2 catalytic reaction enhanced the •OH production in FBR-Fenton/GAC process by 2.7 folds as compared to homogenous Fenton process. Fenton reaction mainly occurred in the upper liquid region and its kinetics for •OH generation significantly diminished by 75% within the first 10 min. GAC/H2O2 reaction took place in the fluidized bed region for continuous •OH generation with a relatively stable rate from 1.21 × 10-6 to 0.60 × 10-6 M/s. Along the ROC treatment with FBR-Fenton/GAC process, the simulated COD degradation rate decreased along the reaction time with 2.05 × 10-6 M/s and 2.93 × 10-7 M/s at 2 min and 60 min, respectively. Faster COD removal was attained in the fluidized bed region due to combining effects of •OH oxidation and GAC adsorption. The overall predicted COD concentration reduced from 122 to 35 mg/L, •OH oxidation and GAC adsorption contributed 59% and 41%, respectively, to the total COD removal.

Long non-coding RNA MIAT promotes non-small cell lung cancer progression by sponging miR-1246.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long non-coding RNA MIAT promotes non-small cell lung cancer progression by sponging miR-1246, by D. Lin, H.-P. Xu, J.-H. Lin, H.-H. Hu, Q. Wang, J. Zhang, published in Eur Rev Med Pharmacol Sci 2019; 23 (13): 5795-5801-DOI: 10.26355/eurrev_201907_18318-PMID: 31298331" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18318.

Genomic origin and EGFR-TKI treatments of pulmonary adenosquamous carcinoma.

BACKGROUND: Adenosquamous carcinoma (ASC) of the lung is a heterogeneous disease that is composed of both adenocarcinoma components (ACC) and squamous cell carcinoma components (SCCC). Their genomic profile, genetic origin, and clinical management remain controversial. PATIENTS AND METHODS: Resected ASC and metastatic tumor in regional lymph nodes (LNs) were collected. The ACC and SCCC were separated by microdissection of primary tumor. The 1021 cancer-related genes were evaluated by next-generation sequencing independently in ACC and SCCC and LNs. Shared and private alterations in the two components were investigated. In addition, genomic profiles of independent cohorts of adenocarcinomas and squamous cell carcinomas were examined for comparison. We have also carried out a retrospective study of ASCs with known EGFR mutation status from 11 hospitals in China for their clinical outcomes. RESULTS: The most frequent alterations in 28 surgically resected ASCs include EGFR (79%), TP53 (68%), MAP3K1 (14%) mutations, EGFR amplifications (32%), and MDM2 amplifications (18%). Twenty-seven patients (96%) had shared variations between ACC and SCCC, and pure SCCC metastases were not found in metastatic LNs among these patients. Only one patient with geographically separated ACC and SCCC had no shared mutations. Inter-component heterogeneity was a common genetic event of ACC and SCCC. The genomic profile of ASC was similar to that of 170 adenocarcinomas, but different from that of 62 squamous cell carcinomas. The incidence of EGFR mutations in the retrospective analysis of 517 ASCs was 51.8%. Among the 129 EGFR-positive patients who received EGFR-TKIs, the objective response rate was 56.6% and the median progression-free survival was 10.1 months (95% confidence interval: 9.0-11.2). CONCLUSIONS: The ACC and SCCC share a monoclonal origin, a majority with genetically inter-component heterogeneity. ASC may represent a subtype of adenocarcinoma with EGFR mutation being the most common genomic anomaly and sharing similar efficacy to EGFR TKI.

[Menopausal symptoms and quality of life of hormone receptor positive breast cancer patients at different endocrine therapy time].

Objective: To explore the menopausal symptoms and quality of life of hormone receptor positive (HR+ ) breast cancer patients at different endocrine therapy time. Methods: The HR+ breast cancer patients who were pathologically confirmed from 2011 to 2017 in the Sichuan Cancer Hospital were divided into three groups according to endocrine therapy time (<12 months, 12~36 months, >36 months) and analyzed by a cross-sectional study. The Menopausal symptoms and quality of life of these patients were measured using the modified Kupperman scale and the functional assessment of cancer therapy-breast cancer (FACT-B) scale. The differences of menopausal symptoms among different time groups and drug groups were analyzed by Chi-square test. The differences of quality of life and the effects of menopausal symptoms on quality of life were tested by covariance and multiple linear regression analyses. Results: The average score of menopausal symptom of 167 patients was 14.5±7.6 and the prevalence rate was 87.4% (146/167). Among all of the menopausal symptoms, the prevalence rate of insomnia was the highest (73.7%, 123/167). Besides insomnia and excitement, hot flashes was more prevalent in selective estrogen receptor modulator (SERM) users (64.8%, 79/122) , while osteoarthritis was more prevalent in aromatase inhibitor (AI) users (62.2%, 28/45). The total score of FACT-B of Patients was 104.5±15.5, and the compliance rate was up to 89.8% (150/167). However, the condition of each dimension was different, the compliance rates of social/ family and functional dimension were lowest, which were 73.0% (122/167) and 50.9% (85/167), respectively. The menopausal symptoms of patients at different time groups were 15.0±1.3, 14.0±6.9, 14.5±7.4, respectively, and the total score of FACT-B of patients at different time groups were 102.7±17.8, 105.0±12.9, 105.6±16.7, respectively, without significant differences (both P>0.05). Multiple linear regression analysis showed that menopausal symptoms impaired the quality of life of SERM users during the endocrine therapeutic period. The standardized regression coefficients of three time groups were -0.67, -0.30, -0.50, respectively, with the lowest effect on 12~36 months group. Conclusion: HR+ breast cancer patients will have a poor function recovery and social/ family return, who need more attention. Menopausal symptoms are common problems during endocrine therapy, and active measures should be taken to improve patients' quality of life.

Long non-coding RNA MIAT promotes non-small cell lung cancer progression by sponging miR-1246.

OBJECTIVE: Recently, long non-coding ribonucleic acids (lncRNAs) have attracted more attention for their roles in tumor progression. The aim of this study was to investigate the exact role of lncRNA MIAT in the progression of non-small cell lung cancer (NSCLC) and to explore the possible underlying mechanism. PATIENTS AND METHODS: MIAT expression in NSCLC tissue samples was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The association between the expression of MIAT and the prognosis of NSCLC patients were explored. Furthermore, the wound healing assay and the transwell assay were conducted in vitro. In addition, the luciferase assay and the RNA immunoprecipitation assay (RIP) were used to elucidate the underlying mechanism. RESULTS: The MIAT expression in NSCLC tissues was significantly higher than that of the corresponding normal tissues. Meanwhile, the MIAT expression was associated with the overall survival time of NSCLC patients. The migration and invasion of cells were significantly promoted after MIAT was over-expressed in vitro. Meanwhile, the cell migration and cell invasion were obviously remarkedly inhibited after MIAT knock-down in vitro. Bioinformatics analysis predicted that microRNA-1246 (miR-1246) was as a novel target for MIAT. The expression of miR-1246 was significantly down-regulated or up-regulated after the overexpression or down-expression of MIAT, respectively. Further mechanism assays showed that miR-1246 was a direct target of MIAT in NSCLC. CONCLUSIONS: MIAT enhanced the NSCLC cell migration and invasion via targeting miR-1246, which might be a potential biomarker in NSCLC.

Whole breast radiotherapy in the isocentric lateral decubitus position: Role of the immobilization device and table on clinical results.

PURPOSE: To evaluate clinical results and the "effect bolus" based on the table design of different linear accelerators in patients with breast cancer treated by previously published whole breast irradiation in the isocentric lateral decubitus position. MATERIAL AND METHODS: We studied 248 consecutive female patients with early stage breast cancer treated by conservative surgery followed by three-dimensional conformal whole breast irradiation in the isocentric lateral decubitus position between January 2013 and February 2014. Radiotherapy was performed on linear accelerators using a Varian. The energy used was 4 and 10MV photons or 6MV photons. All patients were evaluated weekly by the radiation oncologist, acute toxicity was assessed using the NCICTC v 3.0 scale. Late toxicity and cosmetic results were evaluated 18 months after the radiotherapy. Cosmetic results were defined as excellent, good, middle or bad. RESULTS: Among the 248 women included, the median age was 67 years (range: 35-91 years). All received whole breast radiotherapy with boost in 144 patients (58%). One-hundred-twenty patients received normofractionated and 124 patients hypofractionated whole breast radiotherapy. Median follow-up was 18 months. Acute skin toxicity in the whole breast radiotherapy in the isocentric lateral decubitus position was acceptable: there was 47% of grade 1 radiodermatitis, 50% of grade 2 and 3% grade 3 and no grade 4 for normofractionated radiotherapy; 89% of grade 1 dermatitis and 11% of grade 2 for hypofractionated radiotherapy; 89.7% of grade 0-1 dermatitis and 10.3% of grade 2 for the "flash" scheme and did not differ between the three linear accelerators (P=0.2, P=0.9 and P=0.2 respectively for the normofractionated radiotherapy, hypofractionated radiotherapy and the "flash"scheme). Late toxicity was acceptable with 84% of grade 0-1 fibrosis for normofractionated radiotherapy, 94% of patients for hypofractionated radiotherapy and 77% for "flash" scheme and did not differ between the three linear accelerators (P=0.44, P=1 and P=0.22 resp.). Most of patients (81%) had an excellent or a good cosmetic outcome. CONCLUSIONS: Whole breast radiotherapy in the isocentric lateral decubitus position is well tolerated. Clinical results are comparable based on different immobilization device allowed by linear accelerators. Particularly, there was no influence of the couch on skin tolerance and cosmetic results.

Patterns of locoregional failure in women with early-stage breast cancer treated by whole breast irradiation in the lateral isocentric decubitus position: Large-scale single-centre experience.

PURPOSE: The purpose of this study was to evaluate locoregional control and describe the patterns of failure in patients with breast cancer receiving whole breast radiotherapy in the isocentric lateral decubitus position technique. PATIENTS AND METHODS: In a series of 832 consecutive female patients with early-stage breast cancer including invasive and in situ tumours treated by breast-conserving surgery followed by three-dimensional conformal whole breast irradiation in the isocentric lateral decubitus position between 2005 and 2010, all patients who experienced locoregional recurrence were studied. Five-year recurrence-free and overall survival rates were calculated. Regional recurrence mapping patterns were also determined. RESULTS: The median age of this series of 832 women was 61.5 years (range: 29-90 years). Various types of fractionation were used: 50Gy in 25 fractions (17.9%), 66Gy in 33 fractions (50Gy in 25 fractions to breast followed by sequential boost to tumour bed to a total dose 66Gy in 33 fractions.) (46.5%), 40Gy in 15 fractions or 41.6Gy in 13 fractions (26.1%) and 30Gy in 5 fractions (9.5%). With a median follow-up of 6.4 years, only 36 patients experienced locoregional recurrence and no association with the fractionation regimen was identified (P=0.2). In this population of 36 patients, 28 (3.3%) had "in-breast" local recurrences (77.8%), two had local recurrences and regional lymph node recurrence (5.6%), and six had regional lymph node recurrence only (in non-irradiated areas; 16.6%). The median time to recurrence was 50 months. Complete mapping of patterns of recurrences was performed and, in most cases, local recurrences were situated adjacent to the primary tumour bed. Cases of local recurrences presented a significantly lower distant metastasis rate (P<0.001) and had a significantly longer overall survival compared to patients with regional lymph node recurrence (P<0.001). However, multivariate Cox regression analysis showed that the site of recurrence had no significant impact on overall survival (P=0.14). CONCLUSION: The results of this study indicate a low local recurrence rate. Further careful follow-up and recording of recurrences is needed to improve the understanding of patterns of recurrence.

[Locoregional control and patterns of locoregional failure in women with breast cancer treated by highly conformal electron beam irradiation]. / Caractérisation des rechutes locorégionales chez des patientes atteintes d'un cancer du sein traité par irradiation pariétale par électrons : expérience monocentrique de l'institut Curie.

PURPOSE: The purpose of this study was to evaluate locoregional control and describe the patterns of locoregional failure in women with breast cancer irradiated by a previously described post-mastectomy highly conformal electron beam radiotherapy technique. MATERIAL AND METHODS: We included all women irradiated by post-mastectomy highly conformal electron beam radiotherapy technique for non-metastatic breast cancer between 2007 and 2011 in our department. All cases of bilateral breast cancer were excluded. All patients who experienced locoregional recurrence have been studied. Mapping patterns of regional recurrences was also performed and compared with the European Society for Radiotherapy and Oncology (ESTRO) and Radiotherapy Oncology Group (RTOG) guidelines of volume definition and delineation guidelines. RESULTS: With a median follow-up of 64 months (range: 6-102 months), 5-year locoregional recurrence-free and overall survival probabilities were 90 % (95 % confidence interval [95 %CI]: 88.1-92.4) and 90.9 % (95 %CI: 88.9-93), respectively. Among the 796 patients included in the study, 23 patients (2.9 %) presented locoregional recurrences of them only 13 (1.6%) were presented with local recurrence. The majority of them presented aggressive biological features with grade III tumours in 17 patients (74 %) with high mitotic index in 16 cases (70 %) and triple negative tumours in 12 (52 %). Lymphovascular invasion was observed in 11 cases (48 %). In 14 cases the locoregional recurrences were diagnosed at the same time as the metastatic disease whereas 4 patients presented distant metastases secondarily. Locoregional recurrences occurred in 11 cases "in field" although adequate doses and volumes were used and in 12 cases "outfield", out of irradiated volume. Local recurrences occurred in 13 patients with 12 recurrences within the irradiated volumes. Regional recurrences occurred in 13 patients with 15 lymph nodes metastases identified. Four nodal recurrences occurred outside the ESTRO clinical target volume and within the RTOG clinical target volume and two occurred outside both RTOG and ESTRO clinical target volumes. CONCLUSION: In presented series, the local recurrence resulted mostly from of biologic radio resistance whereas regional recurrences were caused by geographical miss. A number of nodal recurrences could occur outside the target volumes defined by ESTRO and RTOG.

[Evaluation of an automatic delineation system for organs at risk and target lymph nodes volumes for patients treated for breast cancer]. / Évaluation d'un logiciel pour la délinéation automatique des organes à risques et des volumes cibles ganglionnaires chez des patientes prises en charge pour un cancer du sein.

PURPOSE: Intensity-modulated radiotherapy needs the strict delineation of target volumes as well as organs at risk and the time used for this procedure is long. The purpose of this study was to evaluate the Workflow Box system (Mirada Medical, UK) for automatic delineation and segmentation for everyday use of organs at risk and lymph nodes delineation in patients treated for early stage breast cancer. MATERIAL AND METHODS: Twenty patients' CT scans in treatment position for their breast cancer radiotherapy were delineated in respect of the ESTRO delineation guidelines to begin the creation of automatic delineation atlas. Then 30 other CT scans were delineated this time by the automatic delineation system and by the radiation oncologist (reference delineation plan). The precision of the delineation was evaluated using the overlap volume index and evaluation of standard deviation (SD). RESULTS: The study of organs at risk has shown that the mean overlap volumes were between 0.49 (SD=0.21) and 0.97 (ET=0.03). Five organs at risk out of nine had overlap volumes at least 0.8. The mean overlap volume for all organs at risk was 0.77 (SD=0.17). The system was less performing for the lymph nodes with a mean overlap volume of 0.43 (SD=0.1) and ranging between 0.23 (SD=0.13) and 0.52 (SD=0.1). The use of this system reduced the delineation time by 40% per patient. CONCLUSIONS: For patients with breast cancer, the system for automatic delineation and segmentation Workflow Box (Mirada Medical, UK) permitted to safely shorten the time for delineation with acceptable organs at risk delineation. Improvement of lymph node volumes is needed. A new evaluation will be realized after using the system in routine practice.

[Predictors of clinically significant bleeding events in bivalirudin-treated Chinese patients with acute myocardial infarction undergoing percutaneous coronary intervention].

Objective: This study was designed to further clarify the independent predictors of clinically significant bleeding events in bivalirudin-treated patients with acute myocardial infarction(AMI) undergoing percutaneous coronary intervention (PCI). Methods: A total of 3 023 AMI patients from 88 centers of China who underwent PCI and received periprocedural bivalirudin treatment between August 2012 and December 2015 were involved in this study.The primary outcome was clinically significant bleeding events defined as the Bleeding Academic Research Consortium(BARC) grades 2-5, with 30 days after PCI.A multivariate Logistic regression model was performed to identify the independent predictors of the primary outcome. Results: Bleeding events occurred in 88(2.9%) patients during the 30-day follow up, with clinically significant bleeding (BARC types 2-5) in 22(0.7%) and BARC types 3-5 in 7(0.2%). Multivariate regression analysis revealed radial access (OR: 0.196, 95%CI: 0.074-0.517, P=0.001) as the independent protector of the significant bleeding events, anemia (OR: 2.956, 95%CI: 1.024-8.528, P=0.045) and eGFR<30 ml/min (OR: 7.860, 95%CI: 1.515- 40.776, P=0.014) as independent risk factors. Conclusions: The rate of clinically significant bleeding complications in Chinese AMI patients undergoing PCI with concomitant use of bivalirudin is low in real-world clinical practice.Radial access is independent protective factor that reduces bleeding events, whereas anemia and eGFR <30 ml/min are independent risk factors that increase bleeding risk.

Effect of intracoronary tirofiban on platelet alpha-granule membrane protein and myocardial perfusion level during emergency percutaneous coronary intervention.

This study aimed to investigate the effect of intracoronary application of tirofiban on platelet alpha-granule membrane protein (GMP-140) and myocardial perfusion levels during emergency percutaneous coronary intervention (PCI). A total of 70 patients who accepted emergency PCI treatment were randomly divided into tirofiban and control groups. We determined GMP-140 and troponin I (cTnI) levels before and 12 h after surgery, as well as N-terminal pro-brain natriuretic peptide levels 1 and 7 days after surgery in the two groups. The results showed that GMP-140 and cTnI levels were significantly (P < 0.01) lower in the tirofiban group than in the control group 12 h after operation (17.99 ± 1.01 vs 24.56 ± 1.96 µg/L and 50.96 ± 2.20 vs 58.69 ± 2.34 ng/mL, respectively). The D-value of the N-terminal pro-brain natriuretic peptide levels between 1 and 7 days after operation was significantly higher in the tirofiban group than in the control group (894.19 ± 90.91 vs 829.50 ± 84.18 pg/mL; P < 0.01). The intracoronary application of tirofiban during emergency PCI clearly reduced the GMP-140 level, inhibited the activation function of platelets, improved myocardial perfusion, and helped recover cardiac function in patients.

Effects of single-dose atorvastatin on interleukin-6, interferon gamma, and myocardial no-reflow in a rabbit model of acute myocardial infarction and reperfusion

The mechanisms of statins relieving the no-reflow phenomenon and the effects of single-dose statins on it are not well known. This study sought to investigate the effects of inflammation on the no-reflow phenomenon in a rabbit model of acute myocardial infarction and reperfusion (AMI/R) and to evaluate the effects of single-dose atorvastatin on inflammation and myocardial no-reflow. Twenty-four New Zealand white male rabbits (5-6 months old) were randomized to three groups of eight: a sham-operated group, an AMI/R group, and an atorvastatin-treated group (10 mg/kg). Animals in the latter two groups were subjected to 4 h of coronary occlusion followed by 2 h of reperfusion. Serum levels of interleukin (IL)-6 were measured by enzyme-linked immunosorbent assay. The expression of interferon gamma (IFN-γ) in normal and infarcted (reflow and no-reflow) myocardial tissue was determined by immunohistochemical methods. The area of no-reflow and necrosis was evaluated pathologically. Levels of serum IL-6 were significantly lower in the atorvastatin group than in the AMI/R group (P<0.01). Expression of IFN-γ in infarcted reflow and no-reflow myocardial tissue was also significantly lower in the atorvastatin group than in the AMI/R group. The mean area of no-reflow [47.01% of ligation area (LA)] was significantly smaller in the atorvastatin group than in the AMI/R group (85.67% of LA; P<0.01). The necrosis area was also significantly smaller in the atorvastatin group (85.94% of LA) than in the AMI/R group (96.56% of LA; P<0.01). In a secondary analysis, rabbits in the atorvastatin and AMI/R groups were divided into two groups based on necrosis area (90% of LA): a small group (<90% of LA) and a large group (>90% of LA). There was no significant difference in the area of no-reflow between the small (61.40% of LA) and large groups (69.87% of LA; P>0.05). Single-dose atorvastatin protected against inflammation and myocardial no-reflow and reduced infarct size during AMI/R in rabbits. No-reflow was not dependent on the reduction of infarct size.

Effects of single-dose atorvastatin on interleukin-6, interferon gamma, and myocardial no-reflow in a rabbit model of acute myocardial infarction and reperfusion.

The mechanisms of statins relieving the no-reflow phenomenon and the effects of single-dose statins on it are not well known. This study sought to investigate the effects of inflammation on the no-reflow phenomenon in a rabbit model of acute myocardial infarction and reperfusion (AMI/R) and to evaluate the effects of single-dose atorvastatin on inflammation and myocardial no-reflow. Twenty-four New Zealand white male rabbits (5-6 months old) were randomized to three groups of eight: a sham-operated group, an AMI/R group, and an atorvastatin-treated group (10 mg/kg). Animals in the latter two groups were subjected to 4 h of coronary occlusion followed by 2 h of reperfusion. Serum levels of interleukin (IL)-6 were measured by enzyme-linked immunosorbent assay. The expression of interferon gamma (IFN-γ) in normal and infarcted (reflow and no-reflow) myocardial tissue was determined by immunohistochemical methods. The area of no-reflow and necrosis was evaluated pathologically. Levels of serum IL-6 were significantly lower in the atorvastatin group than in the AMI/R group (P<0.01). Expression of IFN-γ in infarcted reflow and no-reflow myocardial tissue was also significantly lower in the atorvastatin group than in the AMI/R group. The mean area of no-reflow [47.01% of ligation area (LA)] was significantly smaller in the atorvastatin group than in the AMI/R group (85.67% of LA; P<0.01). The necrosis area was also significantly smaller in the atorvastatin group (85.94% of LA) than in the AMI/R group (96.56% of LA; P<0.01). In a secondary analysis, rabbits in the atorvastatin and AMI/R groups were divided into two groups based on necrosis area (90% of LA): a small group (<90% of LA) and a large group (>90% of LA). There was no significant difference in the area of no-reflow between the small (61.40% of LA) and large groups (69.87% of LA; P>0.05). Single-dose atorvastatin protected against inflammation and myocardial no-reflow and reduced infarct size during AMI/R in rabbits. No-reflow was not dependent on the reduction of infarct size.

Molecular cloning, expression and variation analyses of the dopamine D2 receptor gene in pig breeds in China.

The dopamine D2 receptor (DRD2) is a crucial mediator for normal physiological processes. We cloned the pig DRD2 gene, investigated its distribution in tissues and identified polymorphisms by RT-PCR, quantitative real-time PCR and direct sequencing. Two Yorkshire pigs from Guangdong Academy of Agricultural Sciences (Guangzhou, China) were selected to clone the gene and investigate its expression; 16 individuals from four pig breeds (Yorkshire, Landrace, small-ear spotted, and Xinchang) were used to scan the variations. The two transcripts (DRD2L and DRD2S), obtained through insertion or deletion of exon 5 and part of 3'UTR, were found to encode 444- and 415-amino acid proteins, respectively. The 574-bp indel in 3'UTR comprises five miRNA targeting sites, based on bioinformatics predictions. The pig DRD2 gene expresses predominantly in the pituitary gland, and then in oviducts and the hypothalamus. Both DRD2L and DRD2S mRNA were detected in cerebrum, cerebellum, hypothalamus, pituitary gland, back muscle, oviduct, uterus, and testis tissues; DRD2L was more abundant than DRD2S. The DRD2 gene is located on chromosome 9 and contains seven exons. Sixty-one different sequences were identified in this gene; among seven in the coding region, only one altered the encoded amino acid. These findings will help us understand the functions of the DRD2 gene in pigs.

Polymorphisms associated with egg number at 300 days of age in chickens.

We looked for variations that could be associated with chicken egg number at 300 days of age (EN300) in seven genes of the hypothalamic-pituitary-gonadal axis, including gonadotrophin-releasing hormone-I (GnRH-I), GnRH receptor (GnRHR), neuropeptide Y (NPY), dopamine D2 receptor (DRD2), vasoactive intestinal polypeptide (VIP), VIP receptor-1 (VIPR-1), prolactin (PRL), and the QTL region between 87 and 105 cM of the Z chromosome. Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens, which were obtained from a company located in Guangdong Province of China. The C1704887T of VIPR-1 was found to have a highly significant association with EN300. The T5841629C of DRD2 and the C1715301T of VIPR-1 were significantly associated with EN300. A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300. H1H3 had the highest EN300. Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300. The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300. Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain GRB2-like 2 (SH3GL2) gene. We conclude that five SNPs, including C1704887T and C1715301T of VIPR-1, T5841629C of DRD2, and T32742468C and G32742603A of SH3GL2, would be useful as markers for breeding to increase chicken EN300.

The dopamine D2 receptor gene polymorphisms associated with chicken broodiness.

Chicken broodiness is a polygenic trait controlled by autosomal genes. Prolactin gene is a candidate of great interest in molecular studies of broodiness. However, another candidate dopamine D2 receptor (DRD2) gene has not been studied extensively. The objective of this study was to analyze the genetic effects of the DRD2 gene on chicken broodiness through linkage disequilibrium analyses, tag SNP selection, genetic diversity observation, 2-tailed test, and association analyses. In this study, we assayed 27 variations of this gene in 456 individuals from 6 chicken populations to observe linkage disequilibrium pattern, the tag SNP, and genetic diversity. Among the 6 populations, Taihe Silkies exhibited no characteristic between the square of the correlation coefficient of gene frequencies (r(2)) and physical distance. The other populations including Red Jungle Fowls, Xinghua chickens, Ningdu Sanhuang chickens (NDH), Baier Huang chickens, and Leghorn layers exhibited conspicuous characteristic of decreasing r(2) value over physical distance. Linkage disequilibrium decayed more rapidly in Red Jungle Fowls, Xinghua, and NDH than in Baier Huang and Leghorn layers. Allelic frequencies and genotype distributions in the 5 populations showed that A-38600G, I-38463D, T-32751C, A-16105G, A-6543G, C-6539T, and A+2794G were possibly associated with broodiness. Besides the above 7 sites, another 2 sites that might be associated with broodiness were screened by 2-tailed test. All 9 sites were used for association analyses with broodiness in 644 NDH chickens. A significant association (P < 0.05) was found between A-16105G and broody frequency (%), and the T+619C in intron 1 was significantly associated with duration of broodiness (P < 0.05). These findings suggested that the DRD2 gene should be included in future genetic studies of chicken broodiness and 2 SNP of A-16105G and T+619C might be markers for breeding against broodiness.

Tissue and allelic-specific expression of hsp70 gene in chickens: basal and heat-stress-induced mRNA level quantified with real-time reverse transcriptase polymerase chain reaction.

1. The 70 kDa heat shock proteins (hsp70) are a family of molecular chaperones, which promote protein folding and participate in many cellular functions. The objective of the current research was to investigate the relationship between tissue or allele and the expression of chicken hsp70 under normal growth conditions and during acute heat stress (44 degrees C for 4 h). 2. A total of 279 individuals were genotyped for two single nucleotide polymorphisms (A258G and C276G) in chicken hsp70 gene by single-strand conformation polymorphism (SSCP) analysis. 3. The mRNA abundance of chicken hsp70 genes was evaluated by real-time reverse transcriptase polymerase chain reaction (RT-PCR). The expression of hsp70 gene in the liver (9.83 +/- 0.84, 10(7)) was significantly higher than that in the muscle (4.42 +/- 0.36, 10(7)) under normal growth conditions. However, during acute heat stress, the expression of hsp70 gene in the brain (1.82 +/- 0.25, 10(9)) was the highest and was significantly different from those in the liver (1.08 +/- 0.16, 10(9)) and muscle (1.08 +/- 0.13, 10(9)). 4. The expression of hsp70 among different genotypes or haplotype combinations is quite different under normal and heat-stress conditions. The haplotype H3 (GC) is probably advantageous to improving heat resistance of chickens. 5. The results from the present study indicate that the expression of hsp70 in chickens is affected by heat stress, and is tissue- and allele-dependent.

Cholinergic and dopaminergic amacrine cells differentially express calcium channel subunits in the rat retina.

Immunofluorescence labeling was performed to study the expression of high voltage-activated Ca(2+) channel subunits on rat retinal cholinergic and dopaminergic amacrine cells, which were double labeled with antibodies against choline acetyltransferase and tyrosine hydroxylase, respectively. The alpha(1A) subunit was predominantly expressed on the processes but not on the somata of cholinergic amacrine cells, whereas staining for alpha(1B) and alpha(1E) was observed in both structures of the cells. Immunoreactivity of alpha(1C) and alpha(1D) was not found in the cholinergic amacrine cells. Dopaminergic amacrine cells, on the other hand, exhibited a differential expression pattern of the Ca(2+) channel subunits, with alpha(1A), alpha(1C) and alpha(1E) being expressed on both somata and processes and alpha(1B) predominantly on the processes of the cells. No alpha(1D) labeling was seen. These results suggest that Ca(2+) channel subunits differentially expressed on the cholinergic and dopaminergic amacrine cells may endow these two cell types with different physiological properties.

Signals from cone photoreceptors to L-type horizontal cells are differentially modulated by low calcium in carp retina.

Ca2+ plays crucial roles in both phototransduction and calcium-dependent glutamate release from the photoreceptor terminal. Modulation, by lowering extracellular Ca2+, of red-sensitive (R-) and short wavelength-sensitive (S-) cone-driven light responses of L-type horizontal cells (LHCs) was studied in the isolated superfused carp retina using intracellular recording techniques. Low Ca2+ (nominally Ca2+-free) Ringer's reduced responses of LHCs to both green (500 nm) and red (680 nm) flashes in darkness, with the former being suppressed more substantially than the latter. This differential suppression became more significant when contribution of R-cones to the green-light-induced responses was diminished by a moderate red (680 nm) background light. Application of IBMX, an inhibitor of phosphodiesterase (PDE), increased LHC responses to both red and green flashes equally, resembling the effect of low Ca2+ on phototransduction. In addition, photopic electroretinographic P III responses, reflecting the activity of cones, to red flashes were more potentiated by low Ca2+, compared to those to green flashes, whilst they were both equally potentiated by IBMX. Furthermore, low Ca2+ caused a more pronounced suppression of LHC responses to red flashes than those to green flashes in the presence of IBMX. It is postulated that reduction of LHC responses in low Ca2+ may be due to the 'saturation suppression' caused by the increased glutamate release from the photoreceptor terminal and the differential modulation may reflect a consequence of the dual action of low Ca2+ on the PDE activity in the photoreceptor outer segment and the synaptic strength between cones and LHCs.