Objective:To explore the remodeling of brain function 2 years after anterior cruciate ligament reconstruction (ACLR) and its relationship with functioning and behavior.Methods:Forty-eight volunteers who had received ACLR from the same surgeon were divided into a coping and a non-coping group, each of 17. Another 14 health volunteers formed the control group. Resting-state functional magnetic resonance imaging was used to record blood oxygen level-dependent signals from the members of all three groups in the 0.01 to 0.08Hz band. One-way analysis of variance was applied to the differences in low frequency amplitude (ALFF) observed.Results:The results of multiple comparisons with Gaussian random field theory correction showed that the differences in bilateral putamen ALFF values among the three groups were statistically significant. The signals from the right cerebellar area 8 and the bilateral putamen were significantly stronger among the non-coping group on average. Pearson correlation analysis showed that the ALFF values from the right cerebellar 8 region were positively and significantly more correlated with the symmetry of a subject′s Y balance function test results in the coping group compared with the non-coping group.Conclusions:Subcortical brain function remodeling occurs in ACLR patients returning to exercise after surgery, and a Y-balance function test can indirectly reflect such remodeling. That can provide a basis for designing programs for rehabilitating advanced brain functions.

Objective To explore the difference of brain function remodeling in patients with differ-ent motor ability and record the brain function index of patients returning to exercise 2 years after an-terior cruciate ligament reconstruction(ACLR).Methods Patients undergoing ACLR in year 2017 and 2018(2 years after ACLR)were selected and randomly divided into a return-to-exercise(CP,n=7)group,a non-return-to-exercise(NP,n=7)group.Moreover,8 healthy counterparts of the CP group were chosen into a healthy control(HC)group.Resting state functional magnetic resonance imaging(rs-fMRI)was used to obtain the blood oxygen level dependent signals,and the amplitude of low-frequen-cy fluctuation(ALFF)was computed across the typical band(0.01～0.08 Hz),sub-band Slow-4(0.027～0.073 Hz)and Slow-5(0.01～0.027 Hz).Meanwhile,brain maps were obtained and two-sample t-tests were performed among different groups(P<0.005).Results In the typical frequency band,the average ALFF value was higher in the CP and HC groups than the NP group for the Cerebelum_Crus1,but lower in the CP group than the NP group for the Occipital_Mid,higher in the CP group than the HC group for the Putamen and higher in the NP group than the HC group for the Frontal_Mid_Orb.More-over,in the Slow-4 band,the ALEF level was lower in the CP group than the NP group for the Oc-cipital_Mid,higher in the CP group than the HC group for the Putamen,and higher in the NP group than the HC group for the Frontal_Mid_Orb and cerebellum_Crus2.However,in the Slow-5 band,the ALEF values were higher in the CP group than the NP group for the Occipital_Inf and Precen-tral,but lower for the caudate.In the same band,those values were higher in the CP group than in the HC group for the Cerebellum_Crus1,but lower in the NP group than the HC group for the Cere-bellum_Crus1 and Supp_Motor_Area,and higher in the NP group than the HC group for the Fron-tal_Mid_Orb.Conclusion The patients returning to exercise after ACLR have higher cerebellar remodel-ing and lower visual compensation than those not,and display higher basal ganglia and cerebellar nerve remodeling than healthy controls,suggesting that functional compensation occurs in the former pa-tients.Moreover,Slow-4,Slow-5 and other sub-bands can complement the classical frequency bands and are worthy of further study.

OBJECTIVETo explore the function of a novel nonsense mutation c.1476G>A of ITGB3 gene using an in vitro expression system.

METHODSAn eukaryotic expression vector containing ITGB3 c.1476G>A cDNA was generated by site-directed mutagenesis and transformed into E.coli. Plasmid DNA was extracted and sequenced to confirm the target mutations. Wild-type and mutant recombination plasmids were transfected into Chinese hamster ovarian cancer (CHO) cells by nonliposome method, and the stable expression cells were harvested by G418 screening. The ITGB3 gene mRNA transcription and GPIIIa expression level in CHO cells were detected with real-time quantitative PCR, Western blotting and flow cytometry, respectively.

RESULTSThe eukaryotic expression vectors of wild ITGB3 cDNA and c.1476G>A mutant were successfully constructed. CHO cells with stable expression were obtained after transfection and screening. Compared with the wild-type transfected cells, the amount of CD61 antigen expression was 37% and mRNA transcription level was only 6% in the mutant-transfected cells. Full length GPIIIa protein was found only in the stably wild-type-transfected cells, but not in mutant-transfected cells by Western blotting analysis.

CONCLUSIONThe ITGB3 c.1476G>A mutation can decrease the transcription level and further affect GPIIIa synthesis and CD61 antigen expression.

Animals ; Base Sequence ; Blood Platelets ; cytology ; metabolism ; CHO Cells ; Cloning, Molecular ; Codon, Nonsense ; genetics ; Cricetinae ; Cricetulus ; Humans ; Integrin beta3 ; genetics ; metabolism ; Molecular Sequence Data ; Plasmids ; genetics ; metabolism ; Point Mutation

OBJECTIVETo study the serological characteristics and molecular mechanism for a rare Pk phenotype of the P1Pk blood group system.

METHODSThe blood group of the proband was identified by serological techniques. The coding region and flanking intronic sequences of the β-1,3-N-acetylgalactosyltransferase gene (B3GALANT1) associated with the Pk phenotype were analyzed using polymerase chain reaction sequence-based typing.

RESULTSThe proband was identified as having a rare Pk phenotype including anti-P in her serum. The blood group of her daughter and husband showed a P2 phenotype. The nucleotide sequences of the B3GALANT1 gene of her husband and two randomly-chosen individuals were the same as the reference sequence (GenBank AB050855). Nucleotide position 433 C>T homozygous mutation in the B3GALANT1 was found in the proband, which has resulted in a stop codon at amino acid position 145, which may produce a premature protein capable of decreasing or inhibiting the activity of the β -1,3-N-acetylgalactosyltransferase. The nucleotide position 433 C/T heterozygous in the B3GALANT1 was found in her daughter.

CONCLUSIONThe Pk phenotype resulted from 433 C>T mutation in the B3GALANT1 gene has been identified.

ABO Blood-Group System ; genetics ; Adult ; Base Sequence ; Blood Grouping and Crossmatching ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; N-Acetylgalactosaminyltransferases ; genetics ; Pedigree ; Phenotype ; Point Mutation

OBJECTIVETo explore the molecular basis for an individual with rare p phenotype in the P1Pk blood group system.

METHODSErythrocyte blood group antigens and antibodies in serum were identified in the proband and five family members with a serological method. Coding regions and flanking untranslated regions of the α1,4-galactosyltransferase gene (A4GALT) encoding P1Pk antigens were amplified with polymerase chain reaction and directly sequenced. The haplotypes of A4GALT in the parents of the proband were also analyzed by cloning sequencing.

RESULTSThe proband was found with a rare p phenotype with anti-Tja antibody in his serum by serological method. The other family members all had a common P2 phenotype. The results of DNA sequencing showed that a cytosine was inserted at nucleotide position 1026 to 1029 (1026_1029insC) of both alleles of the A4GALT gene in the proband. The mutation has caused a reading frame shift and formed a mutant protein by extending 92 amino acid residues. The other family members were either heterozygous for the insertion or of the wild type at above position.

CONCLUSIONThe 1026_1029insC mutation of the A4GALT gene is probably responsible for the p phenotype identified for the first time in Chinese population. The individual with the p phenotype possesses anti-Tja antibody.

ABO Blood-Group System ; genetics ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Female ; Frameshift Mutation ; Galactosyltransferases ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutagenesis, Insertional ; Pedigree ; Phenotype ; Young Adult

OBJECTIVETo establish a polymerase chain reaction sequencing-based typing (PCR SBT) method for HLA-DPB1 exons 2 and 3, and to analyze their polymorphisms.

METHODSBased on the sequences of HLA-DPB1 loci, locus-specific primers were designed and applied to amplify the target sequences encompassing the entire exons 2 and 3 of HLA-DPB1. The amplification products were digested by enzymes and directly sequenced in both directions. The genotype was assigned by Assign 3.5+ SBT software.

RESULTSSpecific target fragment was obtained with the PCR amplification, and good quality electropherogram was derived by direct sequencing. Among 242 individuals from Zhejiang Han population, 18 HLA-DPB1 alleles were detected. Alleles with a frequency of > 0.05 have included DPB1*05:01:01/135:01 (0.4112), DPB1*02:01:02 (0.1901), DPB1*04:01:01 (0.1136) and DPB1*02:02 (0.0620). A novel HLA-DPB1*168:01 allele has also been identified. Nine polymorphism sites were founded in the exon 3 region, which included a new SNP site 517 A>T.

CONCLUSIONThe PCR-SBT method for exons 2 and 3 of HLA-DPB1 is reliable, which allowed detection of polymorphisms in exon 3 of the HLA-DPB1 gene.

Alleles ; Exons ; HLA-DP beta-Chains ; genetics ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic

Objective To construct a prokaryotic expression system for major histocompatibility complex class Ⅰ chain-related gene B ( MICB) and to establish an ELISA method for the detection of anti-MICB antibodies in patients with kidney transplantation. Methods The MICB cDNA fragments were ob-tained by RT-PCR with a pair of specific primers. The MICB cDNA and the prokaryotic expression vector pET-28a were digested by two restriction enzymes and ligated to construct the recombinant expression plas-mid pET-28a-MICB. The transformed E. coli BL21 DE3 strains carrying recombinant expression plasmid were induced by IPTG to express MICB protein. The expressed recombinant proteins were identified by Western blot assay and purified by Ni-NTA Spin column. The purified proteins were coupled to ELISA for the detection of anti-MICB antibodies in patients with kidney transplantation. Results Three common MICB fragments contained the exons 2 and 3 were obtained. The recombinant proteins were expressed in E. coli BL21 DE3 strains carrying pET-28a-MICB and successfully purified by the Ni-NTA Spin column. Results of the Western blot assay confirmed that the obtained proteins were the target proteins. The ELISA method was successfully established and used for the detection of anti-MICB antibodies in 24 patients with kidney trans-plantation. The absorbance values indicated that the sensitivities of three recombinant MICB proteins were different. Conclusion The expression system for MICB gene was successfully constructed. The established ELISA for the detection of anti-MICB antibodies would pave the way for further investigation on the correla-tions between MICB protein and transplantation immunity.

[ABSTRACT]OBJECTIVETo investigate the clinic significance of nasal septal swell body by observing and measuring it in the normal and deviated nasal septum on CT images.METHODSThe locations of the nasal septal swell bodies on horizontal CT images in 50 normal subjects and 30 patients with deviated nasal septum were studied, and their length, width and the thicknesses of the mucosa of the both sides were measured. The data were analyzed with SPSS.RESULTSSeptal swell bodies were observed in most of CT images. The swell body was fusiform and located anterior to the middle turbinate, with mean(SD) width of (10.30±1.27) mm and length of (31.35±5.18) mm. There no marked difference in thickness of the nasal septal swell body between two sides of the nasal septum in normal nasal septum, but the thickness of the nasal septal swell body in camber side was thicker than that in the other side of the deviate nasal septum.CONCLUSIONThe shape and location of spetal swell body suggests its potential capacity may be to alter the nasal airflow. Additional study is required for its clinical significance.

Objective To investigate the effects of methylated CpG islands in the promoter region on the expression of killer cell immunoglobulin-like receptor 3DL1 (KIR3DL1).Methods Three voluntary unpaid blood donors carrying high expression allele KIR 3DL1*01502 and three donors carrying low expres-sion allele KIR3DL1*005 were recruited in this study .The nucleotide sequences and the methylated CpG islands in the promoter regions of KIR 3DL1*01502 allele and KIR3DL1*005 allele were analyzed .The NK cells expressing KIR3DL1*01502 and KIR3DL1*005 were respectively treated with 5-aza for the dem-ethylation of CpG islands within the promoters .The expression of KIR3DL1 protein on the surface of NK cells was measured with flow cytometer .Results Two differences at nucleotide sites -65 and -269 were detected within the promoter regions of KIR3DL1*01502 and KIR3DL1*005, resulting in two distinct CpG islands.The CpG islands within the promoter of KIR 3DL1*01502 allele were highly methylated .The ex-pression of KIR3DL1 protein on NK cells which carried KIR 3DL1*01502 allele was significantly increased after the demethylation of CpG islands .However , the treatment of demethylation had no significant effects on the expression KIR3DL1 protein on NK cells harboring KIR3DL1*005 allele.Conclusion The methylated CpG islands within the promoter of KIR 3DL1*01502 allele affected the antigen expression on the surface of NK cells.Different KIR3DL1 alleles might show different mechanisms in regulating antigen expression .

OBJECTIVE: To observe anatomic structure of jugular foramen region by endoscope, to provide anatomic data for avoiding damnification in the surgery. METHOD: We performed the retrolabyrinthine and retrosigmoid endoscopic surgery on 8 fomalin-fixed adult cadaver specimens and observed the structures of jugular foramen by endoscope and compared the different surgeries at the same time. We excised the calvarium and cereburm and exposured and observed the nerves and vessels. Moreover we measured the the distance from internal accoustic pore to glossopharyngeal and analyse the data by SPSS. RESULT: All retrolabyrinthine endoscopic surgeries were performed successfully. Only 4 postsigmoid endoscopic surgeries were performed without damage of cerebellum which is the major obstacles. The distance from internal accoustic pore to glossopharyngeal was(8.26 ± 1.05) mm. About half of posterior inferior cerebellar arteries located to inboard of nerves. CONCLUSION The jugular foramen region endoscopic surgery can be performed successfully by retrolabyrinthine. The "lockhole" technology by retrosigmoid is more difficult for blocking of cerebella. The internal acoustic porus is a fixed structure of the cerebellopontine angleand a perfect landmark to the surgery.
Adult ; Cadaver ; Endoscopy ; Foramen Magnum ; anatomy & histology ; Humans ; Jugular Veins ; anatomy & histology ; Temporal Bone ; anatomy & histology