Identification and Characterization of the ATG8, a Marker of Eimeria tenella Autophagy / Identificação e Caracterização do ATG8, um marcador de autofagia em Eimeria tenella

Abstract Autophagy plays an important role in maintaining cell homeostasis through degradation of denatured proteins and other biological macromolecules. In recent years, many researchers focus on mechanism of autophagy in apicomplexan parasites, but little was known about this process in avian coccidia. In our present study. The cloning, sequencing and characterization of autophagy-related gene (Etatg8) were investigated by quantitative real-time PCR (RT-qPCR), western blotting (WB), indirect immunofluorescence assays (IFAs) and transmission electron microscopy (TEM), respectively. The results have shown 375-bp ORF of Etatg8, encoding a protein of 124 amino acids in E. tenella, the protein structure and properties are similar to other apicomplexan parasites. RT-qPCR revealed Etatg8 gene expression during four developmental stages in E. tenella, but their transcriptional levels were significantly higher at the unsporulated oocysts stage. WB and IFA showed that EtATG8 was lipidated to bind the autophagosome membrane under starvation or rapamycin conditions, and aggregated in the cytoplasm of sporozoites and merozoites, however, the process of autophagosome membrane production can be inhibited by 3-methyladenine. In conclusion, we found that E. tenella has a conserved autophagy mechanism like other apicomplexan parasites, and EtATG8 can be used as a marker for future research on autophagy targeting avian coccidia.

Resumo A autofagia desempenha um papel importante na manutenção da homeostase celular através da degradação de proteínas desnaturadas e outras macromoléculas biológicas. Nos últimos anos, muitos pesquisadores se concentraram no mecanismo da autofagia em parasitas apicomplexos, mas pouco se sabe sobre esse processo na coccidia aviária. No presente estudo, a clonagem, sequenciamento e caracterização de gene relacionado à autofagia Etatg8 foram investigados pela PCR quantitativa em tempo real (RT-qPCR), mancha ocidental (WB), ensaios indiretos de imunofluorescência (IFAs) e microscopia eletrônica de transmissão (TEM), respectivamente. Os resultados mostraram que o gene Etatg8 de E. tenella possui uma ORF de 375 bp, codificando uma proteína de 124 aminoácidos com estrutura e propriedades semelhantes à de outros apicomplexos. RT-qPCR revelou que Etatg8 é expresso durante os quatro estágios de desenvolvimento de E. tenella. Entretanto, seus níveis transcricionais foram significativamente mais elevados na fase de oocisto não esporulados. Os ensaios de manchas ocidental (WB) e de imunofluorescência (IFA) mostraram que a proteína EtATG8 foi lipidada para ligar-se à membrana do autofagossomo sob condições de deficiência nutritiva (em presença de rapamicina) e se agregar no citoplasma de esporozoítas e merozoítas. No entanto, o processo de produção de membrana do autofagossomo pode ser inibido por um inibidor de autofagia (3-meetiladeninatiladenina, 3-MA). Em conclusão, foi demonstrado que E. tenella tem um mecanismo de autofagia conservado, semelhante ao de outros parasitas apicomplexos, e que EtATG8 pode ser usado como um marcador para futuras pesquisas sobre autofagia direcionada à coccidiose aviária.

Efficacy of novel modified Nuss procedure in treatment of pectus excavatum after congenital heart disease operation: A case control study / 中国胸心血管外科临床杂志

@#Objective    To explore the feasibility and efficacy in therapy of pectus excavatum using novel modified Nuss procedure after congenital heart disease (CHD) operation. Methods    Thirty-six children (including 22 males and 14 females with an average age of 4.5±2.2 years ranging 2.8-18.0 years) with pectus excavatum after CHD operation from January 2011 to March 2015 were selected as an observation group. Thirty-eight pectus excavatum children (including 24 males and 14 females with an average age of 4.0±2.5 years ranging 2.8-20.0 years) without CHD from July to September 2013 were selected as a control group. The novel modified Nuss procedure was performed on the patients in both groups. The operation time, intraoperative blood loss, hospital stay as well as postoperative complications between two groups were reviewed and analyzed. Results    In the observation group, the operation time was 50-72 (60.50±3.60) min and hospital stay was 4-6 (4.41±0.80) d. Meanwhile, the intraoperative blood loss was 5-10 (5.82±0.35) ml. In the control group, the operation time was 12-45 (20.15±0.68) min, hospital stay was 4-7 (4.61±0.63) d and the intraoperative blood loss was 3-8 (4.62±0.28) ml. The operation time was significantly longer in the observation group than that in the control group (P<0.05). But there was no significant difference in intraoperative blood loss or hospital stay between the two groups (P>0.05). No cardiac rupture happened in the two groups. Conclusion    The novel modified Nuss procedure is safe and feasible for pectus excavatum after CHD operation with optimal outcomes.

Surgical treatment of 190 adults with pectus excavatum by newly modified orthopedic steel plate / 中国胸心血管外科临床杂志

@#To summarize the clinical experience of treating adults with pectus excavatum by newly modified orthopedic steel plate and Nuss procedure. Methods    The clinical data of 190 adults with pectus excavatum treated by newly modified Nuss procedure in Xinhua Hospital affiliated to Shanghai Jiaotong University School of Medicine from January 2011 to June 2016 were collected. There were 151 males and 39 females aged 21.80±3.96 years ranging from 18 to 45 years. The therapeutic efficacy, Haller index and the lung function index were also analyzed. Results    All patients recovered well after the operation. Two orthopedic steel plates were implanted in 12 patients. Pneumothorax was found in 7 patients postoperatively and 2 of them was treated by puncture extraction. There were 5 patients with poor healing of incision and all of them were healed after the debridement. Other complications such as steel plate shift was found in 2 patients. Both of them recovered after the reoperation. The duration of operation was 36–65 min. The intraoperative blood loss was 5–20 ml. Postoperative hospital stay was 4–7 days. Haller and lung function index improved after the operation (P<0.001). Conclusion    It is effective and safe to treat the pectus excavatum by newly modified orthopedic steel plate and Nuss procedure in adult patients.

Surgical strategy for giant mediastinal mass / 中国胸心血管外科临床杂志

@#Objective    To introduce the surgical and perioperative strategy for giant mediastinal mass. Methods    The clinical data of 21 patients with giant mediastinal mass who underwent surgical treatment in Xinhua Hospital of Shanghai from January 2007 to July 2016 were retrospectively reviewed. There were 14 males and 7 females, with a mean age of 34.62 ± 22.95 years (range: 11 months to 79 years), and mean weight of 58.07±22.24 kg (range: 10.8 to 90.5 kg). Their clinical manifestation, anesthesia methods, surgical treatment and the prognosis were analyzed. Results    The tumor volume ranged from 8 cm×6 cm×6 cm to 25 cm×25 cm×8 cm. For surgical approach, 12 patients received median sternotomy, 5 anterior lateral incision, 1 posterior lateral incision, 2 "L"-shape sternotomy, 1 cervical and thoracic "]"-shape incision. All patients were given mass radical resection, except one patient with two-stage resection. Twelve patients needed other tissues resection besides the single tomor resection. The operation time was 55-480 (207.86±87.67) min, blood loss volume 700 (10-4 000) ml, intraoperative blood transfusion 800 (0-4 100) ml, postoperative mechanical ventilation time 4.75 (0-87) h, postoperative drainage time 3-12 (7.43±2.66) d, the total drainage volume 295-4 940 (1 584.76±1 173.98) ml, average daily drainage volume 62-494 (204.90±105.76) ml, and postoperative hospital stay 7-47 (11.86±8.51) d. The postoperative complications included pericardial effusion in 1 patient, Horner   syndrome in 1, left recurrent laryngeal nerve injury with the left phrenic nerve injury in 1, right phrenic nerve injury in 1 and delayed wound healing in 1. The remaining patients recovered well. All patients were followed up for 1 month to 9 years. Till September 1, 2016, 5 patients died and 2 suffered recurrent tumor. Conclusion    It is safe to perform surgical treatment after comprehensive evaluation of patients with giant mediastinal mass, perioperative mortality is low, and prognosis in patients with benign tumor is good.