TRAF3 of blunt snout bream participates in host innate immune response to pathogenic bacteria via NF-κB signaling pathway.

Tumor necrosis factor receptor-associated factor 3 (TRAF3) is a multifunctional adaptor protein primarily involved in both bacterial defense and antiviral immunity in living organisms. However, the knowledge on TRAF3 in blunt snout bream (Megalobrama amblycephala), a freshwater fish with economic values, remained unclear. In the present study, we identified and characterized successfully Traf3 gene from M. amblycephala (maTraf3). The maTraf3 cDNA contained a 1722 bp open reading frame that encoded a protein of 573 amino acid residues. The deduced amino acid sequence comprised of a RING finger domain, two zinc finger motifs, a coiled-coil region and a MATH domain. Analysis of the transcriptional patterns of maTraf3 revealed that it was ubiquitously distributed in various tissues tested from M. amblycephala, with the abundance of expression in spleen and muscle. Following a challenge with Aeromonas hydrophila and lipopolysaccharide stimulation, the expression of maTraf3 was strongly enhanced at different time points in vitro and in vivo. MaTRAF3 was identified as a cytosolic protein and suggested to form aggregates or be associated with vesicles scattering in the cytoplasm. NF-κB transcription was activated by maTraf3 in reporter assay. The overexpression of maTraf3 produced high levels of pro-inflammatory cytokines such as IL-1ß, IL-6, IL-8 and TNF-α, implying its immune-regulatory role in M. amblycephala. Taken together, our results obtained in this study demonstrated the crucial role of maTraf3 in mediating host innate immune response to pathogen invasion via NF-κB signaling pathway, which might indicate a novel therapeutic approach to combat bacterial infection in fish.

[Curative effect of surgery on moyamoya disease: an analysis of 47 patients].

OBJECTIVE: To evaluate the effect of surgery on 47 patients with moyamoya disease by retrospective analysis.
 Methods: A total of 47 patients with moyamoya disease were enrolled from August, 2010 to According to the improved treatment in August, 2013, all cases were divided into two groups: a pre-improved group and a post-improved group. According to different surgical methods, they were divided into two subgroups: an indirect revascularization subgroup and a combined revascularization subgroup.
 Results: The cerebral ischemia in 77.4% of patients was relieved after the surgery. There was significant difference in outcomes of patients between the pre-improved group and the post-improved group (P<0.05), while there was no significant difference between the pre-improved indirect revascularization subgroup and the pre-improved combined revascularization subgroup. There was also no significant difference between the post-improved indirect revascularization subgroup and the post-improved combined revascularization subgroups (P>0.05).
 Conclusion: Surgical treatment can improve the outcomes of patients with moyamoya disease, but there is no significant difference in surgical effects between indirect and combined revascularization.

Identification and expression analysis on bactericidal permeability-increasing protein/lipopolysaccharide-binding protein of blunt snout bream, Megalobrama amblycephala.

Bactericidal permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP) belong to the lipid transfer protein/lipopolysaccharide-binding protein family and play a critical role in the innate immune response to Gram-negative bacteria. In the present study, a novel BPI/LBP from blunt snout bream, Megalobrama amblycephala (maBPI/LBP) was isolated by RACE techniques. The open reading frame (ORF) of maBPI/LBP gene encoded a polypeptide of 474 amino acids with a putative 18-aa hydrophobic signal peptide. Structurally, the maBPI/LBP showed highly similar to those of BPI/LBPs from invertebrate and teleost, LBPs and BPIs from mammal, which contained an N-terminal BPI/LBP/CETP domain BPI1 with a LPS-binding domain, a C-terminal BPI/LBP/CETP domain BPI2, and proline-rich domain. The homologous identities of deduced amino acid sequences displayed that the maBPI/LBP possessed significant similarity (96.61% and 90.07%) with those of grass carp and common carp, respectively. The recombinant protein of maBPI/LBP showed effectively kill Gram-negative bacteria. The maBPI/LBP gene was expressed in a wide range of normal tested tissues, with the highest expression levels in the kidney. The experiments revealed that the mRNA expression of maBPI/LBP in spleen considerably up-regulated from 2 h to 8 h post LPS stimulation, and peaked rapidly at 2 h (7.40-fold, P < 0.05), which confirmed that maBPI/LBP was the absolute sensitive to LPS stimulation. Furthermore, the level of maBPI/LBP mRNA expression reached the maximum for a second time at 24 h after LPS stimulation. These results suggested that maBPI/LBP was a constitutive and inducible acute-phase protein contributing to the host immune defense against pathogenic bacterial infection in M. amblycephala. This study will further our understanding of the function of BPI/LBP and the molecular mechanism of innate immunity in teleost.

Transdural Collateral Circulation Indicates Cerebral Ischemia in Moyamoya Disease.

BACKGROUND: Moyamoya disease (MMD) is an idiopathic occlusive cerebrovascular disorder. The development of collateral circulation originates from the dural and pial collaterals. Currently, the clinical significance of transdural collateral in MMD has not been established. We sought to study the relationship between transdural collateral circulation and the side of relative cerebral ischemia in MMD. METHODS: Data from MMD patients were collected at Xiangya Hospital from January 2016 to April 2022. A collateral circulation grading system with scores was established, the dominant side of transdural collateral with a higher point. Cerebral perfusion was used to identify the side of relative cerebral ischemia. RESULTS: A total of 102 patients were recruited. Results of digital subtraction angiography showed that 74 (72.5%) patients had transdural collaterals. The transdural collaterals were more common in patients with infarctions than in those with headaches or transient ischemic attacks (P = 0.0074). The dominant side for the formation of transdural collateral circulation was more easily found at the side of relative cerebral ischemia (P < 0.0001). Additionally, the side of the brain with a higher score of transdural collaterals was more likely to experience relative cerebral ischemia (P < 0.0001). There was no significant difference in the formation of transdural collateral circulation between ischemic and hemorrhagic MMD patients. CONCLUSIONS: Transdural collateral circulation was common in MMD patients. The transdural collaterals were associated with the occurrence of infarction. Transdural collaterals were well established on the cerebral ischemic side, which indicated higher ischemic levels in the ipsilateral than contralateral side.

rac-(1R,2S,6R,7R)-4-{[(1E)-(2-Chloro-phen-yl)methyl-idene]amino}-1-isopropyl-7-methyl-4-aza-tricyclo-[5.2.2.0]undec-8-ene-3,5-dione.

The title compound, C(21)H(23)ClN(2)O(2), was synthesized from N-amino-α-terpinene maleimide and 2-chloro-benzaldehyde. There are two independent mol-ecules in the asymmetric unit which are linked via an inter-molecular C-H⋯O hydrogen bond. The crystal studied was found to be a partial merohedral twin, with a 0.74 (7):0.26 (7) domain ratio.

Circular RNA in osteoarthritis: an updated insight into the pathophysiology and therapeutics.

Osteoarthritis (OA) is a common joint disease that mainly results in chronic pain, stiffness and dysfunction in elderly individuals. The molecular mechanisms in the pathogenesis of OA are still unclear, and available treatments are unable to slowdown the development of OA or reverse the tissue damage. Circular RNAs (circRNAs), a novel type of non-coding RNA, are ubiquitous, stable, evolutionally conserved, tissue-specific and functional. An increasing number of studies have revealed that many circRNAs are differentially expressed in OA-affected joint tissues and engage in the pathogenesis of OA by functioning as miRNA sponges. In this review, we briefly introduce the biogenesis, characteristics and functions of circRNAs, and shed light on the important role of circRNAs in the occurrence and progression of OA and their potential diagnostic and therapeutic value in this disease based on the research over the last five years.

Epiphyseal distraction and hybrid reconstruction using polymethyl methacrylate construct combined with free non-vascularized fibular graft in pediatric patients with osteosarcoma around knee: A case report.

BACKGROUND: In children with osteosarcoma around the knee joint without epiphysis involvement, joint-sparing surgery seems to be an ideal way to retain knee joint function. However, there are two points of debate with regard to the technique: How to accurately achieve a safe surgical margin, and how to achieve intercalary reconstruction of the massive bone defect following resection of the tumor. CASE SUMMARY: We present the case of an 8-year-old girl with osteosarcoma of the distal femur without involvement of the epiphysis. Epiphyseal distraction was applied to separate the epiphysis and metaphysis, and this provided a safe surgical margin. The massive bone defect was reconstructed with a custom-made antibiotic-loaded polymethyl methacrylate (PMMA) construct combined with a free non-vascularized fibular graft. Six months after surgery, bone union between the autograft and host bone was confirmed in both the proximal and distal femur by computer tomography (CT) examination. Moreover, considerable callus formation was found around the PMMA construct. After 28 mo of follow-up, there was no sign of recurrence or metastasis. The patient could walk without any aid and carry out her daily life activities satisfactorily. CONCLUSION: In cases of osteosarcoma without epiphysis involvement, epiphyseal distraction can be easily applied to obtain a safe margin. Hybrid reconstruction with an antibiotic-loaded PMMA construct combined with a free non-vascularized fibular graft has the advantages of being easy to manufacture, less time-consuming to place, and less likely to get infected, while also ensuring bone union. Our case provides an alternative technique for biological reconstruction after joint-sparing surgery in patients with osteosarcoma around the knee without epiphyseal involvement.