Objective To explore the expression change of intestinal epithelial tight junction (IJ)protein claudin-1 in mice with fulminant hepatic failure (FHF).Methods FHF was induced with a method that combined intraperitoneal injection of lipopolysaccharide (LPS,10 mg/kg) and D-galactosamine (GalN,800 mg/kg).Control saline (2 ml/kg,ip),LPS (10 mg/kg,ip) and GaIN (800 mg/kg,ip) were also detected.The effect of administration of anti-tumor necrosis factor alpha (TNF-α) IgG antibody (anti-TNF-α IgG,100 μg/per) on the level of TNF-α was assessed before administration of D-galactosamine/lipopolysaccharide.At the 2nd h,6th h,9th h,12th h,24th h after injection in FHF group,the 9th h after injection in control groups and 9th h after injection in anti-TNF-α IgG group,the mice were killed for the collection of large intestine specimens.Claudin-1 was analyzed with immunohistochemistry,Western blotting,and real-time quantitative PCR.Results Tight junction protein claudin-1 was localized along the apical region of the lateral plasma membrane representing the region of tight junctions in surface and crypt epithelial cells.Weakly distributed density of claudin-1 in intestinal mucosa was found in mice with FHF from the 9th h after injection.Compared to saline group,Western blotting analysis demonstrated markedly reduced claudin-1 expression in mice with FHF at the 6th h and 9th h after injection (6th h:0.8600±0.0208 vs 1.0,P ＜0.05; 9th h:0.6633 ±0.0328 vs 1.0,P ＜0.01).Furthermore,the expression of claudin-1 mRNA was markedly reduced at the 6th h,9th h,and 12th h after injection in mice with FHF (6th h:0.3067 ±0.1291 vs 1.0,P ＜0.05; 9th h:0.2233 ±0.1155 vs 1.0,P ＜0.01 ; 12th h:0.5275 ±0.1222 vs 1.0,P ＜0.05).Compared to saline group,no significant difference in claudin-1 expression was found with prophylactic treatment with anti-TNF-α-IgG antibody in mice with FHF at the 9th h after injection (protein:0.9533 ±0.0186 vs 1.0,P ＞0.05; mRNA:0.85 ±0.1437 vs 1.0,P ＞0.05).Conclusions The expression of tight junction protein claudin-1 was reduced at both protein and mRNA levels in intestinal epithelial cells that were induced by TNF-α in mice model of FHF.

Objective To get TDRP1 gene of sterile and fertile boar of the Banna minipig inbred line (BMI), predict its function by bioinformatics analysis, and detect its expression patterns in the fertile boar.Methods Based on the NM_001198925 sequence, we designed specific primers and amplified BMI TDRP1 using RT-PCR method for sequen-cing and bioinformatics analysis.Meanwhile, the expression of TDRP1 in 17 organ tissues ( heart, liver, spleen, lung, kidney, thymus, lymph nodes, skin, duodenum, stomach, cerebrum, cerebellum, testis, epididymis, seminal vesicle, prostate, and bulbourethral gland) of fertile BMI boar and in the testis of sterile and fertile BMI boars was analyzed by semi-quantitative RT-PCR.Results The experiment obtained 680 bp cDNA sequence ( GenBank accession number:KJ186786) of BMI TDRP1, which encodes a protein of 186 amino acids with a predicted molecular weight (Mw) of 20.49 kDa and isoelectric point (pI) 5.86, and no signal peptide.It was a nuclear protein with a probability of 94.1%and had a leucine-rich nuclear export signals.Homology analysis of protein sequences revealed that BMI TDRP1 showed high identi-ty with that of humans, macaca mulatta, mouse and rat.The RT-PCR analysis showed that TDRP1 had a similar expression in the testes of sterile and fertile BMI boars.It was highly abundant in the seminal vesicle and prostate, moderately ex-pressed in cerebellum and testis and weakly expressed in cerebrum and kidney, while undetected in other 11 organ tissues. Conclusions We have cloned TDRP1 complete coding sequence, and found 2 SNPs,showing no difference in sequences and the testis mRNA expression levels between the fertile and sterile BMI boars.The multi-tissue transcription profile shows different expression levels in different organ tissues, being high in the seminal vesicle and prostate.The results of this study provide a foundation for further insight into the role of this gene in spermatogenesis.

Objective:To clone swine leukocyte antigen,class II,DO alpha (SLA-DOA) gene from Banna mini-pig inbred line (BMI) and detect its mRNA expression level in 19 important tissues.Methods:The complete eDNA sequence of SLA-DOA gene was cloned by RT-PCR method from BMI and the mRNA expression pattern in BMI important tissues was examined by semi-quantative RT-PCR method.Nucleotide and protein sequences of SIA-DOA were used to carry out bioinformatics analysis and construct the phylogenetic tree.Results:The eDNA length of BMI SLA-DOA was 1 079 bp,which encoded a protein of 250 amino acids with molecular weight (Mw) 27.81 kD,and isoelectric point (pI) 6.48.Genome structure analysis showed it localized to Sus scrofa chromosomes 7 and consisted of four exons and three introns.Semi-quantitative expression analysis showed that SLA-DOA gene expressed highly in the lymph nodes and stomach;weakly in the heart,skin and duodenum and none in other 14 tissues.Functional bioinformatics analysis indicated that SLA-DOA protein contained one signal poptide,one transmembrane region,three conserved domains,four O-GlcNAc glycosylation sites,18 potential phosphorylation sites and to be located in the cytoplasm with 94.1％ certainty.Phylogenetic analysis demonstrated that BMI (pig) had the closest relationship with cattle.Conclusion:This study have successfully cloned the SLA-DOA gene of Banna mini-pig inbred line,performed bioinformatics analysis and tissue expression profile analysis.It will provide a basis for the studies of BMI xenotransplantation.

In order to remove the endotoxin from the blood of endotoxemia patients, we prepared a new adsorbent with heparin space arm and polymyxin B (PMB) ligand. The carrier of chloromethyl polystyrene resin was activated and heparin space arm was grafted, and then PMB ligand was immobilized onto adsorbent with glutaraldehyde. We employed in vitro FITC-lipopolysaccharide (FITC-LPS) static adsorption to characterize the adsorption properties on the adsorbent, and conducted in vitro lipopolysaccharide (LPS) static adsorption to measure quantitavely the adsorption capacity and rate, and then evaluated the blood compatibility. The in vitro static adsorption indicated that the adsorbent had the removal rate of LPS above 70% with the adsorption equilibrium time for 2 hours. Blood compatibility experiment showed that the adsorbent had little negative effects on blood cells and plasma protein, and their adsorption rates were less than 10% for hemocytes and 20% for plasma protein respectively. This adsorbent exhibited high selectivity, high adsorption capacity and good biocompatibility, and presented a promising clinical application in the treatment of endotoxemia.
Adsorption ; Endotoxemia ; therapy ; Endotoxins ; isolation & purification ; Hemofiltration ; instrumentation ; methods ; Heparin ; chemistry ; Humans ; Ion Exchange Resins ; chemistry ; Ligands ; Polymyxin B ; chemistry ; Sorption Detoxification ; methods

Objective:To investigate the effect of concentrated growth factor (CGF) on proliferation and differentiation in Beagle adipose-derived stem cells (ADSCs).Methods:ADSCs were isolated from adipose tissue of healthy Beagles and cultured.The multidirectional differentiation potential of ADSCs was identified.The ADSCs were assigned to a CGF group and a control group.The rate of proliferation was analyzed by CCK-8 assay.The osteogenic differentiation capability was detected by ALP staining after the osteoinduction.Bone formationrelated gene expression was detected by RT-PCR.Results:CGF promoted the proliferation of ADSCs in vitro.ADSCs in the CGF group showed higher level of ALP activity than that in the control group (P＜0.05).CGF stimulated the expression of the genes associated with osteogenesis,such as Col-I and Runx2.Conclusion:CGF can promote the proliferation and osteogenic differentiation in ADSCs in vitro.

Accurate segmentation of pediatric echocardiograms is a challenging task, because significant heart-size changes with age and faster heart rate lead to more blurred boundaries on cardiac ultrasound images compared with adults. To address these problems, a dual decoder network model combining channel attention and scale attention is proposed in this paper. Firstly, an attention-guided decoder with deep supervision strategy is used to obtain attention maps for the ventricular regions. Then, the generated ventricular attention is fed back to multiple layers of the network through skip connections to adjust the feature weights generated by the encoder and highlight the left and right ventricular areas. Finally, a scale attention module and a channel attention module are utilized to enhance the edge features of the left and right ventricles. The experimental results demonstrate that the proposed method in this paper achieves an average Dice coefficient of 90.63% in acquired bilateral ventricular segmentation dataset, which is better than some conventional and state-of-the-art methods in the field of medical image segmentation. More importantly, the method has a more accurate effect in segmenting the edge of the ventricle. The results of this paper can provide a new solution for pediatric echocardiographic bilateral ventricular segmentation and subsequent auxiliary diagnosis of congenital heart disease.
Adult ; Humans ; Child ; Heart Ventricles/diagnostic imaging* ; Echocardiography ; Image Processing, Computer-Assisted