Objective:To construct a fusion protein of extracellular domain peptide fragment of muscle specific kinase ( MuSK) and fluorescent protein mCherry ,and used as antigen in the detection of antibodies against MuSK ( MuSKAb ) in the sera of patients with myasthenia gravis ( MG).Methods:The mCherry gene was amplified by PCR from vector pRSET-B and cloned into pGEM-T Easy Vector,and furthermore, cloned into Eukaryotic expression vector pMT /BiP/V5-His ( MuSK), which contains MuSK extracellular domain 22-452 amino acid peptide fragment gene to construct the fluorescent fusion protein gene MuSK -mCherry.The recombinant vector was subsequently transfected into drosophila S 2 cells for expression.The expressed fusion proteins were verified in confocal mi-croscope ,and used as antigen in the detection of MuSKAb in sera of MG patents in fluorescence immunoprecipitation test .Results:The fluorescent fusion protein MuSK-mCherry was successfully constructed and expressed.The MuSKAb in sera of patents with MG could be detected in fluorescence immunoprecipitation test using the constructed MuSK-mCherry fusion protein as antigen.Conclusion: It is available to use the constructed fluorescent fusion protein MuSK-mCherry as antigen in fluorescence immunoprecipitation test for the detection of MuSKAb in sera of patents with MG.

Objective To establish a zebrafish model of thrombosis induced by three kinds of inducers and observe the anti?thrombotic effect of a Chinese traditional medicine, Guanxinning tablet ( GXN) . Methods The zebrafish models of thrombosis was induced by using 1?5μmol/L phenyl hydrazine, 80μmol/L arachidonic acid and 5 mg/L ponatinib, re?spectively, and were treated with various concentration of GXN, clopidogrel or asprin. The thrombus in the tail vein was observed under microscope, Erythrocytes in the zebrafish heart were stained with o?dianisidine and the erythrocyte staining intensity was assessed with a NIS?Elements DTM image analyzer, and the anti?thrombolic effect of GXN was calculated. Results Venous thrombus was significantly increased and the staining intensities of erythrocytes in the heart were signifi?cantly decreased after induction by phenyl hydrazine, arachidonic acid or Ponatinib ( P <0?001 ) , respectively. At the same time, GXN showed an incresing anti?thrombolic effect in the zebrafish models (P<0?001) in a dose?effect manner, with a IC50 of GXN of 44?32 mg/L,138?5 mg/L and 459?5 mg/L, respectively. Conclusions The zebrafish models of thrombosis are successfully established by phenyl hydrazine, arachidonic acid or Ponatinib, respectively, by different for?mation mechanisms. GXN has been shown to have an anti?thrombosis effect, probably, by multiple target effects.

Objective:To investigate the association of single nucleotide polymorphisms (SNPs) of receptor-associated protein at the synapse ( rapsyn ) with myasthenia gravis ( MG ).Methods: The genomic DNA was extracted from peripheral blood cells , sampled from 132 patients with MG and 153 control individuals.The 8 exons of rapsyn gene were amplified by PCR ,then the products of PCR sequenced directly.Each sequence was compared with wild-type rapsyn gene , and the association between mutation and clinical symptoms of MG analysed.Results:No mutation was found in the exons 1,2,4,5,6,7,and 8 of rapsyn gene both in MG patients and control group compared with the wild-type rapsyn gene.However,a new SNP,L222R[CTG>CGG(2)] or T665G,was found in exon-3.The allele and genotype frequencies of SNP L 222R met Hardy-Weinberg genetic equilibrium (P>0.05),indicating the group repre-sentativeness.The allele frequencies of G were not statistically different between patient and control groups ( P>0.05 ).There were differences in the 3 genotypes TT , TG and GG between patient ( 42.4% vs 48.5% vs 9.1%) and control ( 49.0% vs 33.3% vs 17.6%) groups ( P<0.05 ).The genotype frequencies of GG were statistically higher in control group than that in patient group , showing a recessive model of inheritance.Conclusion: The SNPs in the rapsyn gene are associated with MG in this study.L222R ( T665 G) is a new SNP found and allele G might be a protective factor for MG.

Objective To analyze the correlation between the molecular subtypes of non-mass enhancement (NME)breast cancer and MRI findings.Methods A retrospective analysis of MRI images and clinicopathological data of 62 patients with NME breast cancer who received 3.0T breast MRI multimodal scan from January 201 5 to June 201 9 was conducted.Molecular subtypes were classified by immunohistochemical staining.The differences between MRI findings of NME breast cancer and molecular subtypes were compared by ch-i square test and Fisher exact probability method,and the correlation between them was analyzed by cross contingency table.Results The differences of molecular subtypes of NME breast cancer in the internal enhancement and peritumoral edema were statistically significant (P＜0.05),while the differences in the TIC,lesion distribution type and peritumoral vascular were not statistically significant (P＞0.05).Conclusion Luminal A type was less prone to peritumoral edema in NME breast cancer.Luminal B type was mainly heterogeneous enhancement. HER-2 overexpression type and triple negative type are mainly clustered ring enhancement,and prone to peritumoraledema.Different molecular subtypes of NME breast cancer have certain MRI characteristics,which is expected to provide a non-invasive evaluation for the preoperative treatment of NME breast cancer.

Objective: To explore the advantages and clinical efficacy of free chimeric perforator flap based on the descending branch of circumflex femoral artery applied to tongue reconstruction after advanced tongue cancer resection. Methods: From October 2013 to December 2018, 57 cases received tongue and oral base reconstruction surgeries using the descending branch of circumflex femoral artery chimeric perforator flap, including 39 males and 18 females, ranged from 20 to 76 years old. And all cases were with stage T3 and T4 tongue cancers, including 35 cases of squamous cell carcinoma, 7 cases of low differentiation cancer, 5 cases of oncosarcoma, and 10 cases of adenoid cystic carcinoma. The tongue was reconstructed by using perforator flap and muscle flap to fill the dead space at the oral floor. The artery anastomoses with the superior thyroid artery or facial artery in the receiving area, and the vein anastomoses with the internal jugular vein in the receiving area. The shape, function and local complications of the reconstructed tongue were observed after operation. Results: Of 57 cases, only one case had partial necrosis of flap, while other 56 cases with chimeric perforator flap survived. Postoperative gastric tube and tracheal cannula were removed in all patients, no cases with oral fistula. All donor sites were sutured in one stage. Postoperative radiotherapy was performed in 41 of the patients. All patients were followed up for 3 to 60 months (average of 20.7 months), with satisfactory esthetic and functional results in reconstructed tongues. Only linear scars were left in the donor areas of the legs, and no lower limb dysfunction was observed. Conclusions The descending branch of circumflex femoral artery chimeric perforator flap can used for repairing simultaneously the defects of both tongue and oral base. It is helpful to avoid the occurrence of oral fistula and to provide the reconstructed tongue with a good function. It is a good choice to use the descending branch of circumflex femoral artery chimeric perforator flap for tongue reconstruction after resection of advanced tongue cancer resection.