Effect of Xijiao Dihuang decoction on microRNA expression in liver tissue of septic mice / 中华急诊医学杂志

Objective:To explore the mechanism of Xijiao Dihuang Ddecoction (XJDHT) against sepsis-induced liver injury based on transcriptomics.Methods:Sixty C57BL/6 mice were randomly (random number) divided into the sepsis group, sepsis treatment with XJDHT and control group, with 20 mice in each group. The sepsis mouse model was established by intraperitoneal (i.p.) injection of lipopolysaccharide (LPS). The control group was intraperitoneally injected with the same amount of normal saline. The sepsis treatment with XJDHT group was injected with XJDHT (crude drug 187.5 mg) twice a day 2 days before modeling. After modeling, gastric feeding was continued twice a day, while the control group and sepsis group were gavaged with the same amount of normal saline. At 72 h after LPS intervention, 9 mice in each group were randomly selected. After anesthesia, part of the liver were taken for small RNA and RNA sequencing and analysis, and part of the liver were taken for pathological examination.Results:XJDHT could improve the histopathological changes of liver in septic mice, and alleviate some abnormally expressed microRNAs (mmu-mir-292a-5p, mmu-mir-871-3p, mmu-mir-653-5p, mmu-mir-293-5p, mmu-mir-155-3p, mmu-mir-346-5p, mmu-mir-187-5p, mmu-mir-3090-3p) and their target genes.Conclusions:XJDHT can reduce the liver histopathological changes in septic mice, and its mechanism may be related to XJDHT regulating the expression of important key genes of liver of sepsis like mmu-mir-187-5p and its target genes such as ADAM8, irak3 and PFKFB3

Analysis of genetic variants in a pedigree affected with hereditary multiple osteochondroma / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a pedigree affected with hereditary multiple osteochondroma (HMO).@*METHODS@#Peripheral blood samples were collected from the proband and members of his pedigree with informed consent. Following extraction of genomic DNA, all coding exons and flanking intronic sequences (-10 bp) of the EXT1 and EXT2 genes were subjected to targeted capture and next generation sequencing (NGS). Suspected variant was verified by Sanger sequencing.@*RESULTS@#A heterozygous nonsense variant (c.1911C>A) was found in exon 10 of the EXT1 gene in the proband and his affected father but not in a healthy sister and normal controls. The variant was classified as a pathogenic based on the guidelines of the American College of Medical Genetics and Genomics (PVS1+PM2+PP1). Bioinformatic analysis predicted that the c.1911C>A variant may be disease-causing via nonsense-mediated mRNA decay and anomalous splicing.@*CONCLUSION@#The c.1911C>A variant probably underlay the disease in this pedigree. Discovery of this variant enriched the variant spectrum of HMO.

Analysis of a multiple osteochondroma case caused by novel splice mutation (c.1164+1G to A) of EXT1 gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect potential mutation of EXT1 gene in a pedigree affected with multiple osteochondroma and explore its pathogenic mechanism.</p><p><b>METHODS</b>The coding regions and their flanking sequences of the EXT1/EXT2 genes were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified by excluding possible single nucleotide polymorphisms and bioinformatics analysis. Transcripts of the EXT1 gene in the proband were analyzed by TA clone-sequencing, with its abundance compared with that of healthy controls.</p><p><b>RESULTS</b>DNA sequencing has identified in the proband a novel heterozygous point mutation (c.1164+1G to A) at the 5'splice sites of intron 3 of the EXT1 gene. The same mutation was not found in the healthy controls. Bioinformatics analysis indicated that the mutation is highly conserved and can lead to skipping of exon 3 or aberrant splicing. TA clone-sequencing indicated that the numbers of transcripts with skipping of exon 3 has significantly increased in the proband (< 0.05) compared with the controls.</p><p><b>CONCLUSION</b>The c.1164+1G to A mutation has resulted in skipping of exon 3 in a proportion of EXT1 gene transcripts. As the result, the number of transcripts with tumor suppressing function is relatively reduced and has ultimately led to the tumors.</p>

Gene-expression profiling of spleen in sepsis rat models / 中华急诊医学杂志

Objective To find out the differences in gene expression of spleen tissue in septic rats by using DNA microarrays.Methods Thirty male Wistar rats were randomly (random number) and equally divided into control group and sepsis group,and septic rat model was induced by cecal ligation puncture (CLP).The rats of control group were only subjected to a simulated operation without CLP.Gene expression profiles were studied by using RatRef-12 gene chip.Rat gene expression profile was showed by using microarray to detect the changes in gene expression pattern of rat spleen tissue after CLP.And subsequently,by using relevant computer software to screen and analyze,the comparison of differences in gene expression between the sepsis group and control group was made.Results Of 22 523 genes,205 differential genes were found between sepsis group and control group,accounting for 0.910％.Among them 98 genes showed up-regulation,with 48 known functional genes,and 107 genes showed down-regulation,with 64 known functional genes.The function of such different genes were associated mainly with apoptosis,inflammation and energy metabolism of spleen cells.Conclusions Splenic dysfunction may be attibuted to the abnormal expression of relevant genes subjected to apoptosis,inflammation and alteration of energy metabolism.It may be the cause of immunosuppression in the later stage of sepsis.

The effect of Ulinastatin on gene expression of septic rat's spleen tissue / 中华普通外科杂志

Objective To explore the effect of UTI (Ulinastatin) preconditioning on gene expression profiles of spleen tissue in septic rats by DNA microarray technology. Methods Forty-five male Wistar rats were equally divided into sham group,sepsis group and UTI group by means of random number table.In UTI group the rats were treated with intramuscular injection of UTI( 105 U/kg) one hour before cecal ligation and puncture.In sepsis group and sham group intramuscular balanced solution (5 ml/kg) was given.Cecal ligation and puncture was used to reproduce septic rat model. Gene expression profile was studied by using RatRef-12 Rat gene expression profile microarray to detect the changes in gene expression pattern of rat spleen tissue after cecal ligation and puncture.Then using related computer software was used to screen and analyze the relationship between the Sepsis/UTI group and sham group. Results In 22 523 genes,205 differential genes were found between sepsis group and sham group,accounting for 0.910％.Among them 98 genes were up-regulated,with 48 known functional genes and 32 genes only showed in this group;107 genes were down-regulated,with 64 known functional genes and 34 genes only showed in it.197 differential genes were found in UTI group and sham group,accounting for 0.875％.Among them 114 genes were up-regulated,with 35 known functional genes and 19 genes only showed in this group; 83 genes were down-regulated,with 49 known functional genes and 19 genes only showed in it. Conclusions Abnormal expression of genes in the spleen tissue of rats with sepsis owing to excessive inflammation and immune suppression were partly relieved by UTI preconditioning.UTI pretects spleen at genetic level.