改良医用胶黏合假疝囊预防腹腔镜全腹膜外直疝修补术后血清肿临床观察

目的探讨应用改良医用胶黏合假疝囊技术预防腹腔镜全腹膜外直疝修补术后血清肿的可行性。方法纳入2017年4月至2019年5月西安市中心医院普一科收治的36例腹股沟直疝患者。所有患者采用TEP手术。术中直疝假疝囊均给予改良医用胶黏合预防术后血清肿。结果36例患者,其中男31例,女5例;年龄31~82岁,平均年龄67.5岁;体质量指数17.6~31.5 kg/m^(2);平均体质量指数24.6 kg/m^(2)。其中Ⅱ型疝13例,Ⅲ型疝23例,均为单侧腹股沟直疝;其中右侧疝25例,左侧疝11例。所有手术均顺利完成,无中转腹腔镜经腹腹膜前疝修补术病例,无感染等严重并发症。手术时间36.6~53.2 min,平均用时44.5 min,其中1例因游离弓状线外侧缘时分破腹膜,术中用3-0可吸收线缝合,因而手术时间延长。术中出血1~5 ml,平均3 ml;术后24~36 h下床,平均下床时间30 h;术后1~3 d出院,平均2 d;住院花费1135.42~1269.74元,平均1189.42元。术后经视觉模拟评分进行评估,所有患者术后1d评分为2~5分,平均3.5分。其中2例分别于术后1周和2周出现血清肿,经腹股沟超声检查证实,血清肿发生率为5.56%,1例因较小,未特殊处理,随访3个月后自行消失,1例给予2次局部抽液,3个月后消失。余患者均未出现明显血清肿。结论改良医用胶黏合假疝囊技术预防直疝TEP术后血清肿的技术,方法简便易行,效果可靠,值得推广及进一步研究。

Fish red blood cells express immune genes and responses

The erythrocytes or red blood cells(RBCs)and leucocytes or white blood cells(WBCs)are the most abundant cell types in the circulatory system of vertebrates.The RBCs are responsible for gas exchange while WBCs are solely involved with immunity.In this study,we present evidence that fish RBCs despite their respiratory role they also express immune-related genes and responses in the teleost fish,Nile tilapia(Oreochromis niloticus).Fluorescent staining identified the presence of numerous mitochondria and lysosomes in tilapia RBCs.RNA sequencing of tilapia RBCs and WBCs led to the assembly and annotation of 20.876 transcripts of which 1017 and 3628 transcripts were specific to RBCs and WBCs,respectively.Pairwise sequence comparisons identified 3251 transcripts that were significantly differential expressed of which 707(21.75%)were up-regulated and 2544(78.25%)down-regulated in RBCs when compared to WBCs.809 transcripts were assigned to 16 immune system pathways,21 and 163 transcripts were specifically expressed in tilapia RBCs and WBCs respectively,and 23 and 233 transcripts were highly expressed in RBCs and WBCs respectively.Expression of 7 immune genes(interferon regulatory factors(irf)1e5 and 9 and tlr3)in both WBCs and RBCs was validated by RT-PCR analysis.Quantitative real-time PCR revealed that all genes were up-regulated in poly(I:C)-challenged WBCs and irf(1,3,4,and 9)and tlr3 were also significantly up-regulated in poly(I:C)-challenged RBCs.Therefore,fish RBCs expressed immunity genes and responses and they may play a complementary role in vertebrate immunity.

microRNA expression profile of fish erythrocytes

MicroRNAs(miRNAs)are short noncoding sequences that can regulate the expression of target genes.Erythrocytes,also called red blood cells(RBCs),are the largest proportion of cells in blood.Erythrocytes deliver oxygen in the body of vertebrates and contribute to the body’s immune function.Illumina sequencing technology was used to analyze pooled RNA samples isolated from erythrocytes,in order to characterize their gene expression profile and increase the range of miRNAs identified in Nile tilapia erythrocytes.This study identified conserved(n=309)and novel(n=194)miRNAs.Quantitative reverse-transcription polymerase chain reaction and next-generation sequencing results indicated that miR-451 was the most abundant in RBCs,followed by let-7a-1,miR-2188 and miR-144.Moreover,the tissue expression mode of these highly expressed miRNAs was characterized.Results show that miR-451,miR-2188,and miR-144 were all highly expressed in RBCs but were low abundance in other tissues.miR-1 and miR-10b were also abundantly expressed in muscle,while miR-122 was abundantly expressed in the liver.let-7a-1,miR-10b,and miR-1 were significantly higher in the testis than in the ovary.The tissue distribution of miRNAs implies that these noncoding sequences have a potential role in regulating tissue differentiation or maintaining tissue characteristics.The results of a dual-luciferase reporter assay indicated that miR-144 and miR-10b inhibited the expression of the meningioma expressed antigen 5 gene in vitro.This study contributes to the Nile tilapia miRNA database and reveals the profile of erythrocyte miRNA expression.

Transcriptome analysis of gill from Lateolabrax maculatus and aqp3 gene expression

The Lateolabrax maculatus is an important marine fish in China that can adapt to a wide range of water salinities.The gill is the main organ involved in fish osmoregulation and in this study,a cDNA library of the L.maculatus gill was constructed to understand gill function in Lateolabrax maculatus and it adaptation to salinity.A total raw data of 5.3G base pairs was obtained,followed removal of low-quality and linker DNA sequences and it yielded a total of 23,038,590 clean reads.After splicing and clustering,31,976 unigenes were obtained.The average transcript length was 1327 bp and N50 was 2057 bp.In comparisons with other tissues analysis of the gill transcriptome revealed a high level of relative expression of apolipoprotein E,CD74,keratin,type I cytoskeleton 13,carbonic anhydrase,betaine-homocysteine S-methyltransferase,and aquaporin 3(AQP3)genes.The L.maculatus aqp3(Lm-aqp3)gene was specific for the gills Lm-aqp3 and this gene has a typical transmembrane structure of six alpha helices and a water channel domain.Phylogenetic analysis indicated that Lm-aqp3 clusters with the homologue genes from Labrus bergylta and Dicentrarchus labrax.Quantitative RT-PCR confirmed that Lm-aqp3 was highly abundant in the gills and low expressed in other tissues(stomach,liver,brain,spleen and testis).Analysis of Lm-aqp3 in freshwater aquaculture fish revealed that gene expression was significantly statistical higher than in seawater(20 ppt)fish,suggesting that Lm-aqp3 may play an important role in water metabolism and osmotic pressure regulation in L.maculatus.

Hemocytes transcriptome profile of the Chinese mitten crab(Eriocheir sinensis)

The Chinese mitten crab(Eriocheir sinensis)possesses an open circulatory system,in which hemolymph moves through interconnected sinuses or spaces surrounding organs called hemocoels.The hemocytes are classified into hyalinocytes,semigranulocytes,and granulocytes;and limited transcriptomics research is available.In this study,the transcriptome of the crab E.sinensis hemocytes was characterized.A total of 14,380,229 clean reads representing a total of 4.31 Gb nucleotides dataset were produced.A total of 67,047 contigs were obtained and 12.49%(8375)and 9.74%(6533)of the contigs were matched to data publicly available from the GenBank nr nucleotide and Uniprot databases,respectively.Among these contigs,4344 contigs belong to three categories of Gene Ontology,126 contigs to 21 subcategories of KEGG,and 4962 contigs to 25 categories of the COG database.A total of 508,379 and 345 transcripts of the E.sinensis showed>40%sequence similarity with transcripts expressed in the vertebrate blood cells from tilapia(Oreochromis niloticus),Chinese softshell turtle(Trionyx sinensis)and chicken(Gallus gallus)respectively.A total of 53,077 single nucleotide polymorphisms were identified and 9912 SSRs for microsatellite mining were found.The most frequent repeat motifs detected were dinucleotide repeats,accounting for 37.52%of the total repeats.Our data provides an important gene resource for the exploitation of molecular marker-assisted breeding,immune mechanisms and conservation of germplasm in the crab E.sinensis.