Histone deacetylase 3 promotes innate antiviral immunity through deacetylation of TBK1

TANK-binding kinase 1 (TBK1), a core kinase of antiviral pathways, activates the production of interferons (IFNs). It has been reported that deacetylation activates TBK1; however, the precise mechanism still remains to be uncovered. We show here that during the early stage of viral infection, the acetylation of TBK1 was increased, and the acetylation of TBK1 at Lys241 enhanced the recruitment of IRF3 to TBK1. HDAC3 directly deacetylated TBK1 at Lys241 and Lys692, which resulted in the activation of TBK1. Deacetylation at Lys241 and Lys692 was critical for the kinase activity and dimerization of TBK1 respectively. Using knockout cell lines and transgenic mice, we confirmed that a HDAC3 null mutant exhibited enhanced susceptibility to viral challenge via impaired production of type I IFNs. Furthermore, activated TBK1 phosphorylated HDAC3, which promoted the deacetylation activity of HDAC3 and formed a feedback loop. In this study, we illustrated the roles the acetylated and deacetylated forms of TBK1 play in antiviral innate responses and clarified the post-translational modulations involved in the interaction between TBK1 and HDAC3.

The screening and identification of internalized nanobody against EpCAM / 药学学报

Epithelial cell adhesion molecule (EpCAM) is a popular target for cancer therapy. In this research, 3 nanobodies with high specificity and endocytosis activity against EpCAM were developed, which provides a basis for the study of immunotoxin based on EpCAM. In our preliminary experiments, we have immunized a camel with EpCAM-Fc antigen and constructed a high-quality phage display library. Seventeen nanobodies with different complementarity determining region (CDR) 3 sequences have been screened after 3 rounds of biopanning by phage display technology. The animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of Fudan University School of Pharmacy. After purification, 7 nanobodies showed high cell binding activity by fluorescent activated cell sorting (FACS) identification. Furthermore, 3 nanobodies presented high endocytosis activity based on FACS and laser confocal microscopy, which also showed high affinity to EpCAM measured by ForteBio. According to this study, we aimed to provide a novel alternative approach to the EpCAM-targeted therapy and to provide guidance for the study of nanobody based immunotoxins for other targets.

Construction and screening of phage display library for TIM-3 nanobody / 药学学报

Immunotherapy is a new strategy for cancer treatment that has the potential to treat all types of cancer. T cell immunoglobulin and mucin-domain-containing molecule-3 (TIM-3) is a key negative regulator of T cell activation. TIM-3 blockage using anti-TIM-3 monoclonal antibody therapy has a great appeal and special advantages. Nanobodies, derived from heavy chain fragment in camelid animals, are now proving clinical values in the development of antibody drugs. In this study, we have immunized camel with TIM-3 antigens and then constructed phage display library. Moreover, 29 nanobodies with different complementarity-determining regions sequences have been screened from the phage display library by phage display technology. In addition, we successfully constructed the cell line stably expressing TIM-3, and screened 10 TIM-3 nanobodies with high specificity and high affinity using flow cytometry. Our study will lay the foundation for the future screening and development of anti-TIM-3 whole humanized functional nanobody.

Treatment of femoral shaft fractures by closed reduction and interlocking intramedullary nailing / 中国骨伤

<p><b>OBJECTIVE</b>To discuss the clinical effect of closed reduction and interlocking intramedullary nailing in the treatment of femoral shaft fracture.</p><p><b>METHODS</b>From March 2006 to December 2011,103 patients with femoral shaft fracture were treated by closed reduction and interlocking intramedullary nailing including 76 males and 27 females with an average age of 36 years old ranging from 19 to 55 years old. According to AO classification,there were 64 cases with type A,27 with type B, 12 with type C. Thirteen cases were open fractures including 5 cases with Gustilo type I , 8 with Gustilo type II . The time of bone healing were observed after operation, the knee function recovery was evaluated by HSS scoring standard at 1 year after operation.</p><p><b>RESULTS</b>The intraoperative complications included femoral neck fracture in 1 case and proximal femoral fracture in 1 case,both of the patients were treated with reconstructive intramedullary interlocking nail and the fractures healed postoperatively. One patient was suffered from common peroneal nerve injury,which were fully recovered at 4 months later after medical treatment. All the patients were followed up from 12 to 28 months (averaged 22 months). All of the fractures were healed well and the average healing time was 3 to 9 months (averaged 5 months). All the hip joints were recovered to normal function. The average HSS was 90.89±5.06 at 1 year after operation.</p><p><b>CONCLUSION</b>Interlocking intramedullary nailing is the preferred treatment for patients with femoral shaft fracture. Carefully operating and closed reduction can reduce the complications.</p>

ESD versus EMR for precancerous lesions and early cancer at gastroesophageal junction / 中华消化内镜杂志

ObjectiveTo evaluate the efficacy and safety of endoscopic submucosal dissection (ESD) for precancerous lesions and early cancer at gastroesophageal junction (GEJ) by comparing endoscopic mucosal resection (EMR) with ESD.MethodsData of patients with GEJ precancerous lesions or early cancer,who received EMR ( n =51 ) or ESD ( n =28) were reviewed to compared the en bloc resection rate,R0 resection rate,operation time,complication and recurrence rate between 2 methods.ResultsEn blcc resection and R0 resection rates of ESD group (92.9％,78.6％ respectively) were significantly higher than those of EMR group (45.1％,43.1％ respectively).Local recurrence rate in ESD group (3.6％,1/28) was significantly lower than that of EMR group ( 19.6％ ).Complications including perforation,delayed hemorrhage,stricture were not significantly different between EMR and ESD groups.Mean operation time of ESD group (64.3 ±27.1 min) was significantly longer than that of EMR group (27.6 ± 14.1 min)(P ＜0.05).ConclusionESD,with a higher cure rate and en bloc rate and a lower local recurrence rate,is superior to EMR for precancerous lesions and early cancer at GEJ.