Modified bilateral carotid artery ligation to establish a vascular dementia rat model and investigate changes in cerebral blood flow and effects on angiogenesis-related proteins / 中国实验动物学报

Objective Apply modified bilateral carotid artery ligation to establish a VD rat model to observe changes in cerebral blood flow and expression of angiogenic proteins.Methods Thirty-six SD male rats were randomly divided into a sham group(n = 18)and model group(n = 18).In the sham group,only the bilateral carotid artery was isolated without ligation,whereas in the model group,the bilateral carotid artery was ligated to establish the VD model.The Morris water maze behavior test was applied before and 14 days after modeling.Variation in cerebral blood flow was detected by laser speckle contrast imaging.Protein expression of HIF-1α,VEGF,and HO-1 was detected by Western Blot.IL-4 and IL-10 contents were measured by ELISA.Results At 14 days after modeling,escape latency was significantly prolonged and the frequency of crossing the platform had significantly decreased in the model group compared with the sham group(P<0.05).At 2 hours,3 days,and 7 days after modeling,cerebral blood flow in the model group was significantly lower than that in the sham group(P<0.05).At 14 and 21 days after modeling,no significant difference was found in cerebral blood flow between sham and model groups(P>0.05).In the model group,cerebral blood flow was decreased to a minimum at 2 hours after modeling(P<0.05)and then began to recover.The peak of recovery occurred at 3～7 days after modeling and returned to the level before modeling on day 14 after modeling.At postoperative day 21,expression of HIF-1α,VEGF,and HO-1 proteins in the hippocampus of the model group was increased remarkably(P<0.05)and the serum contents of IL-4 and IL-10 in the model group were significantly increased compared with those in the sham group(P<0.05).Conclusions The variation in cerebral blood flow in the VD rat model established by the modified bilateral carotid artery ligation was dependent on time.At postoperative day 21,HIF-1α,VEGF,and HO-1 in the hippocampus were increased significantly,which was accompanied by increased levels of IL-4 and IL-10.

Serological feature and molecular mechanism for a case with A307 subgroup / 中华医学遗传学杂志

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.

Application and optimization of CRISPR/Cas system in bacteria / 生物工程学报

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.

Identification and pedigree analysis of ABO subgroup B303 / 中华医学遗传学杂志

<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>

Clinical Value of Serum PIVKA-lI and AFP Detection for Primary Hepatocellular Carcinoma / 现代检验医学杂志

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.