ABSTRACT
OBJECTIVE@#To explore the serological feature and molecular mechanism for a case with A307 subgroup of the ABO blood group system.@*METHODS@#Serological assay was carried out to determine the ABO blood group of the proband and his family members. Genotypes for exons 1 to 7 of the ABO gene were determined with sequence-specific primer polymerase chain reaction (SSP-PCR) and direct sequencing. The impact of the variant on the stability of alpha-1,3-N-acetylgalactosaminyltransferase (GTA) was predicted through construction of a 3D molecular model.@*RESULTS@#The proband, his brother and daughter were diagnosed with Aend phenotype by serological analysis. Their ABO genotype was determined as A307/O02, with heterozygous c.467C>T (p.P156L) and c.745C>T (p.R249W) variants identified in exon 7 of the ABO gene. Molecular modeling suggested that the p.R249W variant may alter the number of hydrogen bonds between the amino acids. The protein was predicted to have a decreased Δ Δ G value of thermodynamic stability.@*CONCLUSION@#The p.R249W variant may give rise to the A307 subgroup by reducing the stability of the GTA enzyme, leading to serological features of Aend phenotype.
ABSTRACT
Clustered regular interspaced short palindromic repeats (CRISPR) system has been widely used in recent years. Compared with traditional genome editing technology, CRISPR/Cas system has notable advantages, including high editing efficiency, high specificity, low cost and the convenience for manipulation. Type Ⅱ and Ⅴ CRISPR/Cas system only requires a single Cas9 protein or a single Cpf1 protein as effector nucleases for cutting double-stranded DNA, developed as genome editing tools. At present, CRISPR/Cas9 technology has been successfully applied to the genome editing of eukaryotes such as zebrafish, mice and human cells, whereas limited progress has been made in the genome editing of bacteria. In our review, we describe CRISPR/Cas system, its mechanism and summarize the optimization and progress of genome editing in bacteria.
Subject(s)
Animals , Humans , Mice , Bacteria , CRISPR-Cas Systems , Endonucleases , Gene EditingABSTRACT
<p><b>OBJECTIVE</b>To explore the molecular mechanism of a case of ABO discrepancies based on the results of blood group serology.</p><p><b>METHODS</b>Five cases of the two-generation pedigrees were analyzed. ABO genotypes were determined using serological tests. DNA sequence analysis was performed on exon 6, exon 7 and intron 3 of the 5 cases to confirm the genotypes of a proband with B subgroup and 4 family members.</p><p><b>RESULTS</b>There were 3 cases of subgroup AB3 and 1 case of subgroup B3 among the 5 family members. The genotypes were identified as A102/B303 and O02/B303, respectively. B303 differed from B101 by intron 3 point mutation (intron3 + 5G>A).</p><p><b>CONCLUSION</b>The point mutation of intron 3 (intron 3+5G>A) is specific in B303.</p>
ABSTRACT
Objective To discuss the clinical value of protein induced by vitaminK absence antagonist-Ⅱ (PIVKA-Ⅱ)and al-pha-fetoprotein (AFP)in diagnosing primary hepatocellular carcinoma (PHC).Methods There were 178 samples from in-patients in Fuzhou Infectious Disease Hospital,including 54 patients with PHC,39 patients with liver cirrhosis,55 patients with hepatitis and 30 cases of healthy.Serum levels of PIVKA-II and AFP levels were detected by LUMI-PULSEG1200 au-tomatic immunity analyzer and Abbott automatic immunity analyzer respectively,and the difference between the levels was compared.Analyzed the areas under the receiver operating characteristic curves (ROC-AUC)and compared the sensitivity and specificity of single PIVKA-II or AFP assay,and the combined detection of PHC.Results The serum level of PIVKA-Ⅱ in hepatocellular carcinoma group was 274 mAU/ml,which was higher than that in liver cirrhosis group (23 mAU/ml), chronic hepatitis group (26 mAU/ml)and healthy group (21 mAU/ml)(P<0.001),and the levels of AFP in PHC group was 84.0 ng/ml,which was higher than that in liver cirrhosis (21.78 ng/ml)and healthy groups (2.8 ng/ml).But it was not statistically significant (P=0.585)compared with those in the chronic hepatitis group (66.8 ng/ml),the results of re-ceiver operating characteristic (ROC)curve showed that the area under the curve of PIVKA-Ⅱ was 0.776,higher than the AFP (0.649),(Z=2.262,P=0.023 7).Serum PIVKA-Ⅱ (≥40 mAU/ml)had a sensitivity of 78.52% and a specificity of 76.23% in the diagnosis of PHC,While serum AFP (≥10 mg/ml)had a sensitivity of 77.78% and a specificity of 34.64%in the diagnosis of PHC.A combination of serum levels of PIVKA-Ⅱ and AFP could increase the sensitivity in the diagnosis of PHC (vs PIVKA-Ⅱ,P=0.031;vs AFP,P=0.016)and specificity (vs PIVKA-Ⅱ,P=0.004;vs AFP,P=0.001).Con-clusion Serum PIVKA-Ⅱ have high clinical application values in diagnosing PHC.A combination of serum levels of PIV-KA-Ⅱ and AFP could increase the sensitivity and specificity in diagnosis of PHC.