Construction of a Spink5 conditional knockout mouse model and analysis of its phenotype / 中华皮肤科杂志

Objective:To construct a serine protease inhibitor Kazal type-5 (Spink5) conditional knockout mouse model, and to identify its phenotype.Methods:B cell-specific Spink5 conditional knockout mice of genotype Mb1 cre/+Spink5 floxp/floxp were constructed by using clustered regularly interspaced short palindromic repeats (CRISPR) /CRISPR-associated protein 9 (Cas9) technology, and served as the knockout group. Mice of genotype Mb1 +/+Spink5 floxp/floxp served as the control group. The mice of genotype Mb1 cre/+Spink5 floxp/floxp or Mb1 +/+Spink5 floxp/floxp were sacrificed when they were 4 to 6 weeks old, splenic mononuclear cells were isolated, and B lymphocytes and non-B lymphocytes were sorted by flow cytometry and fluorescence-activated cell sorting. Genotype identification was performed by PCR, and protein expression of lymphoepithelial Kazal-type-related inhibitor (LEKTI) was determined by Western blot analysis. Skin tissues were resected from the mice, and subjected to hematoxylin-eosin staining for measuring the epidermal thickness. Immunofluorescence staining was performed to determine fluorescence intensity of LEKTI protein in the mouse skin tissues. Paired t test or two-independent-sample t test was used for comparisons between groups. Results:Genotype identification results demonstrated that the stable B lymphocyte-specific Spink5 conditional knockout mouse model was successfully constructed. Western blot analysis revealed that the relative protein expression of LEKTI in the B lymphocytes in the knockout group was 0.01 ± 0.02, which was significantly lower than that in the non-B lymphocytes in the knockout group (0.66 ± 0.11, t = 9.99, P < 0.001) , and that in the B lymphocytes in the control group (1.08 ± 0.13, t = 13.78, P < 0.001) . Among 39 mice in the knockout group, 4 presented with dry skin and scattered scaly hypertrophic maculopapules. The epidermal thickness of the lesional skin tissues in the knockout group was 90.42 ± 21.31 μm, significantly higher than that of the non-lesional skin tissues in the knockout group (29.71 ± 3.63 μm, t = 5.05, P = 0.002) and that of normal skin tissues in the control group (12.42 ± 2.21 μm, t = 6.74, P < 0.001) . Immunofluorescence staining showed no significant difference in the fluorescence intensity of LEKTI protein among the lesional skin tissues (46.21 ± 1.21) , non-lesional skin tissues (46.62 ± 2.13) in the knockout group and normal skin tissues in the control group (47.69 ± 1.71, P > 0.05) . Conclusion:The B lymphocyte-specific Spink5 conditional knockout mouse model was successfully constructed, which provides a basis for further exploring mechanisms underlying skin barrier defects and immune dysfunction in Netherton syndrome.

Blood-cerebrospinal fluid (CSF) barrier dysfunction means reduced CSF flow not barrier leakage - conclusions from CSF protein data / Disfunção da barreira hemato-liquórica significa redução do fluxo de líquido cefalorraquidiano, e não quebra de barreira - conclusões a partir de estudos sobre as proteínas do LCR

ABSTRACT Background: Increased concentrations of serum proteins in cerebrospinal fluid (CSF) are interpreted as blood-CSF barrier dysfunction. Frequently used interpretations such as barrier leakage, disruption or breakdown contradict CSF protein data, which suggest a reduced CSF flow rate as the cause. Results: Even the severest barrier dysfunctions do not change the molecular size-dependent selectivity or the interindividual variation of the protein transfer across barriers. Serum protein concentrations in lumbar CSF increase with hyperbolic functions, but the levels of proteins that do not pass the barrier remain constant (brain proteins) or increase linearly (leptomeningal proteins). All CSF protein dynamics above and below a lumbar blockade can also be explained, independent of their barrier passage, by a reduced caudally directed flow. Local accumulation of gadolinium in multiple sclerosis (MS) is now understood as due to reduced bulk flow elimination by interstitial fluid (ISF). Nonlinear change of the steady state in barrier dysfunction and along normal rostro-caudal gradients supports the diffusion/flow model and contradicts obstructions of diffusion pathways. Regardless of the cause of the disease, pathophysiological flow blockages are found in bacterial meningitis, leukemia, meningeal carcinomatosis, Guillain-Barré syndrome, MS and experimental allergic encephalomyelitis. In humans, the fortyfold higher albumin concentrations in early fetal development decrease later with maturation of the arachnoid villi, i.e., with beginning CSF outflow, which contradicts a relevant outflow to the lymphatic system. Respiration- and heartbeat-dependent oscillations do not disturb net direction of CSF flow. Conclusion: Blood-CSF and blood-brain barrier dysfunctions are an expression of reduced CSF or ISF flow rate.

RESUMO Introdução: Concentrações aumentadas de proteínas séricas no líquido cefalorraquidiano são interpretadas como disfunção da barreira (hemato-liquórica) sanguínea do LCR. Interpretações frequentemente usadas, como vazamento de barreira (quebra ou rompimento de barreira), rompimento ou quebra, contradiz os dados de proteína do LCR, que sugerem uma taxa de fluxo reduzida do LCR como a causa. Resultados: Mesmo as disfunções de barreira mais graves não alteram a seletividade dependente do tamanho molecular nem a variação interindividual da transferência de proteína através de barreiras. As concentrações de proteínas séricas no LCR lombar aumentam com as funções hiperbólicas, mas as proteínas que não passam a barreira permanecem constantes (proteínas do cérebro) ou aumentam linearmente (proteínas leptomeningeais). Toda a dinâmica das proteínas do LCR acima e abaixo de um bloqueio lombar também pode ser explicada, independente de sua passagem pela barreira, por um fluxo caudal reduzido. O acúmulo local de gadolínio na esclerose múltipla (EM) é agora entendido como decorrente da redução da eliminação do bulk flow pelo fluido intersticial (FIS). A mudança não linear do estado estacionário na disfunção da barreira e ao longo dos gradientes rostro-caudais normais apoia o modelo de difusão/fluxo e contradiz as obstruções das vias de difusão. Independentemente da causa da doença, os bloqueios fisiopatológicos do fluxo são encontrados na meningite bacteriana, leucemia, carcinomatose meníngea, síndrome de Guillain-Barré, EM e encefalomielite alérgica experimental. Em humanos, as concentrações de albumina quarenta vezes mais altas no desenvolvimento fetal inicial diminuem tarde com a maturação das vilosidades aracnoides, isto é, com o início do fluxo de LCR, o que contradiz um fluxo relevante para o sistema linfático. As oscilações dependentes da respiração e do batimento cardíaco não perturbam a direção do fluxo do LCR. Conclusão: As disfunções das barreiras hemato-liquórica e hemato-encefálica são uma expressão da redução da taxa de fluxo do LCR ou FIS.

Relationship between airway epithelial barrier dysfunction and allergic asthma / 国际儿科学杂志

Airway epithelial barrier dysfunction has been observed in allergic asthma patients.Inhaled allergens have shown a disruptive effect on the airway epithelial barrier.Airway barrier related genetic variation and epigenetics are also involved in the airway epithelial barrier dysfunction.A lot of studies believe that airway epithelial barrier dysfunction underlie the development of asthma.The study of the relationship between allergic asthma and airway epithelial barrier function will be helpful to understand the pathogenesis of allergic asthma and provide new ideas for the treatment of allergic asthma.

Advances in pathogenesis of intestinal barrier dysfunction in children / 国际儿科学杂志

Intestinal barrier dysfunction refers to the damage or even atrophy of the intestinal mucosa,disorder of intestinal microbial population and increased intestinal permeability caused by a variety of factors,resulting in the insertion of bacterial and/or endotoxin translocation into other tissues and/or blood circulation,induction and/or aggravation systemic multiple organ dysfunction and inflammatory response.The destruction of the intestinal barrier is associated with many gastrointestinal disorders,bur also accompanied by parenteral pathological conditions,such as allergic diseases.Therefore,maintaining a healthy intestinal barrier is critical to children.At present,there are relatively few studies on the intestinal barrier dysfunction in children.This paper reviews the recent progress in the pathogenesis of intestinal barrier in children.

Current status on the intestinal barrier dysfunction in HIV infection / 肠外与肠内营养

Highly active antiretroviral therapy (HAART) can effectively suppress thehuman immunodeficiency virus (HIV) replication andsignificantly reduce morbidity and mortality of HIV-infectedpatients,which howevercan't completely remove the virus,and eventually progressinto chronic viral-infection disease.Chronic HIVinfection destroys host immune system,leading to intestinal barrier damage,intestinal mucosal dysfunction,microbial translocation,and further accelerates the disease progress.The reconstruction of intestinalmicroflora balance and improvement of intestinal mucosa function areessential to reestablish the host immune system.This paper will review the current research advanceson intestinal barrier damage of HIV infection and gut-target therapy of AIDS.The aim is to provide valid evidences for further research targeting improvement of treatment strategiesandreduction of morbidity and mortality in HIV infection.

Effects of hydrogen-rich medium on lipopolysaccharide-induced intestinal epithelial barrier dysfunction of human colon carcinoma cells / 中华危重病急救医学

Objective To investigate the effects of hydrogen-rich medium on lipopolysaccharide (LPS)-induced intestinal epithelial barrier dysfunction of human intestinal epithelial (Caco2) cells. Methods Caco2 cells (passages 28-35) were purchased from the Cell Bank of the Shanghai Institute of Cell Biology, Chinese Academy of Sciences in Shanghai, China, and they were cultured in Dulbecco minimum essential medium (DMEM) containing 20% fetal bovine serum. These cells were randomly divided into four groups: control group (group A), hydrogen-rich medium group (group B), LPS group (group C) and LPS + hydrogen-rich medium group (group D). Cells were cultured with normal medium in group A and group C or with hydrogen-rich medium in group B and group D. Meanwhile, 1 g/L LPS was simultaneously added into group C and group D, while an equivalent volume of normal saline was added into group A and group B instead. In vitro intestinal epithelial models were reproduced with monolayer filter-grown Caco2 and intestinal epithelium. The trans-epithelial electrical resistance (TEER) in models of each group was measured at different incubation times (0, 3, 6, 12, 24 and 48 hours). Cell viability and cytotoxicity were assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) release assay, respectively, after incubation for 24 hours. The expression levels of claudin-1 and occludin were respectively determined at 6, 12 and 24 hours of incubation by Western Blot assay. The morphological structure of claudin-1 and occludin was respectively observed after incubation for 24 hours with immunofluorescence staining. Results There was no statistical significance in variables between group A and group B. Compared with group A, it was shown that TEER was time-dependently decreased in groups C and D after 6 hours. Compared with group C, TEER in group D was increased after 6 hours. Compared with group A, the cell viability was significantly reduced in group C [(67.2±7.9)% vs. (100.0±0.0)%, P < 0.05] and cell injury was obvious [LDH release rate: (38.5±2.1)% vs. (1.2±0.3)%, P < 0.05]; the expression levels of claudin-1 and occludin at 6, 12, 24 hours were significantly down-regulated [claudin-1 (gray value): 0.351±0.079, 0.272±0.075, 0.190±0.049 vs. 0.518±0.030; occludin (gray value): 0.416±0.044, 0.290±0.062, 0.226±0.019 vs. 0.602±0.038, all P < 0.05], and the structure of claudin-1 and occludin were profoundly disrupted. Compared with group C, it was shown that the cell viability was significantly increased in group D [(88.8±7.4)% vs. (67.2±7.9)%, P < 0.05] and cell injury was significantly abated [LDH release rate: (16.4±4.3)% vs. (38.5±2.1)%, P < 0.05]; the expression levels of claudin-1 and occludin were significantly up-regulated at 24 hours [claudin-1 (gray value): 0.428±0.046 vs. 0.190±0.049, occludin (gray value): 0.466±0.071 vs. 0.226±0.019, both P < 0.05]; the disrupted structures of claudin-1 and occludin were partially recovered. Conclusion Hydrogen-rich medium can effectively attenuate LPS-induced dysfunction of intestinal epithelial barrier in human Caco2 cells by ameliorating cell viability as well as regulating claudin-1 and occludin expression and structure.

Intestinal barrier dysfunction and necrotizing enterocolitis / 国际儿科学杂志

Necrotizing enterocolitis ( NEC) is one of the most serious diseases of digestive system dur-ing neonatal period,which is one of the main cause of premature death. The components which maintain the in-testinal barrier function of newborns,especially the premature infants,are always underdeveloped,and easily to be damaged. Thus,the formation of tight junctions between epithelial cells is broken,the early intestinal peristal-sis established delayed,and the secretion of sIgA is reduced. These pathogenic factors induce serious complica-tions,such as intestinal barrier dysfunction,bacterial translocation and sepsis. Hypoxia ischemia,inflammation, infection can either cause intestinal mechanical barrier damage. The delay of micro ecological barrier establish-ment,the immature of immune barriers,intestinal microcirculation dysfunction are all involved in the occurrence of NEC. In addition,miRNA also plays an important role in the regulation of intestinal epithelial cell differentia-tion,structure and barrier function. Pathological changes of NEC are the result of intestinal barrier dysfunction, and the injury of intestinal barrier function will aggravate NEC pathological changes. Therefore, understanding the role of intestinal barrier dysfunction in the pathogenesis of NEC may improve the prevention and treatment of NEC.

The protection of carbachol on intestinal barrier function in patients with trauma / 中国中西医结合急救杂志

Objective To observe the protection of carbachol on intestinal barrier function in patients with trauma. Methods A prospective randomized controlled trial was conducted. Seventy patients after trauma with a definite diagnosis of multiple organ dysfunction syndrome(MODS)from Department of Critical Care Medicine in Hebei United University Affiliated Hospital were included. According to random number table,the patients were divided into a carbachol treatment group(37 cases)and a mosapride citrate treatment group(33 cases),and all the patients in the two groups were treated by antibacterial drugs,supportive agents for organ function,surgery, etc symptomatic treatment. Based on the conventional treatment,in the carbachol treatment group,carbachol was administered through a stomach tube at the dose of 0.2 mg/kg,twice a day,and the dose was doubled if no exhaust or defecation persisted for 3 days after treatment,while in the mosapride group,mosapride citrate was given at the dose of 5 mg once and thrice a day,the therapeutic course of both groups being 7 days. On the 1st,3rd,5th, 7th day after admission,peripheral venous fasting blood in early morning was collected,the activity of diamine oxidase(DAO),expression rates of CD11b+and CD18+in polymorphonuclear neutrophil(PMN),contents of tumour necrosis factor-α(TNF-α)and interleukin-10(IL-10) were detected,and the clinical curative effects were observed. Results Compared to the mosapride citrate treatment group,the total effective rate was significantly higher in the carbachol treatment group on the 7th day after treatment〔70.3%(26/37)vs. 45.5%(15/33),P<0.05〕. The activity of DAO,expression rates of CD11b+and CD18+in PMN,contents of TNF-αand IL-10 in the carbachol treatment group were decreased with the extension of time,and reached valley values on the 7th day,the differences were statistically significant in the comparisons with those in mosapride citrate treatment group at the same time point〔DAO(mg/L):3.21±0.52 vs. 3.93±0.51,CD11b+:(14.89±2.16)% vs.(28.92±1.59)%,CD18+:(53.67±2.44)% vs. (72.46±4.08)%, TNF-α(ng/L):111.44±16.42 vs. 129.73±18.74, IL-10(ng/L):67.71±38.83 vs. 121.45±40.23,all P<0.05〕. At the various time points,the above indexes had no obvious changes in mosapride citrate treatment group. Conclusion Carbachol can ameliorate the ischemic/reperfusion(I/R)injury in patients with intestinal barrier dysfunction after trauma,decrease the release of inflammatory cytokines in vivo,and promote peristalsis of intestinal tract,therefore carbachol has clinical value of protecting intestinal barrier function.

Pathophysiology of psoriasis.

Psoriasis is a chronic inflammatory papulosquamous disease characterized by multiple remissions and relapses. For long, it was believed to be primarily a disorder of keratinization. However, the successful use of traditional immunosupressants and newer immunomodulatory agents in the treatment of psoriasis led to the belief that psoriasis is primarily a disease of Th1 cell immune dysregulation. Recent developments have brought up several new findings such as the role of Th17 cells and evidence of skin barrier dsysfunction in psoriasis, akin to atopic dermatitis. The present review aims to focus on these new developments and explain the pathogenesis of psoriasis on the basis of currently available information.

Pathophysiology of psoriasis.

Psoriasis is a chronic inflammatory papulosquamous disease characterized by multiple remissions and relapses. For long, it was believed to be primarily a disorder of keratinization. However, the successful use of traditional immunosupressants and newer immunomodulatory agents in the treatment of psoriasis led to the belief that psoriasis is primarily a disease of Th1 cell immune dysregulation. Recent developments have brought up several new findings such as the role of Th17 cells and evidence of skin barrier dsysfunction in psoriasis, akin to atopic dermatitis. The present review aims to focus on these new developments and explain the pathogenesis of psoriasis on the basis of currently available information.

Clinical use of a ceramide-based moisturizer for treating dogs with atopic dermatitis

In humans, skin barrier dysfunction is thought to be responsible for enhanced penetration of allergens. Similar to conditions seen in humans, canine atopic dermatitis (CAD) is characterized by derangement of corneocytes and disorganization of intercellular lipids in the stratum corenum (SC) with decreased ceramide levels. This study was designed to evaluate the effects of a moisturizer containing ceramide on dogs with CAD. Dogs (n = 20, 3~8 years old) with mild to moderate clinical signs were recruited and applied a moisturizer containing ceramide for 4 weeks. Transepidermal water loss (TEWL), skin hydration, pruritus index for canine atopic dermatitis (PICAD) scores, and canine atopic dermatitis extent and severity index (CADESI) scores of all dogs were evaluated. Skin samples from five dogs were also examined with transmission electron microscopy (TEM) using ruthenium tetroxide. TEWL, PICAD, and CADESI values decreased (p < 0.05) and skin hydration increased dramatically over time (p < 0.05). Electron micrographs showed that the skin barrier of all five dogs was partially restored (p < 0.05). In conclusion, these results demonstrated that moisturizer containing ceramide was effective for treating skin barrier dysfunction and CAD symptoms.

Clinical study of octreotide in the treatment of intestinal endotoxemia / 中国医师进修杂志

Objective To observe the clinical effect of continuous intravenous pumping of octreotide in the treatment of intestinal endotoxemia.Methods Eighty patients with intestinal obstruction and non-surgical treatment were divided into group Ⅰ with 34 cases who received conventional-treatment and group Ⅱ with 46 cases who received conventional-treatment combined with octreotide 24 h continuous intravenous pumping.White blood cell count ( WBC ),diamine oxidase (DAO),D-lactic acid (D-LA) and endotoxin were detected before treatment and at 24 h,48 h,4 d after treatment.Results The content of WBC,DAO,D-LA and endotoxin in two groups all reached peak at 48 h after treatment.The difference of the content of WBC,DAO,D-LA and endotoxin between two groups had no statistical significance at 24 h after treatment (P ＞ 0.05).The content of WBC,DAO,D-LA and endotoxin of group Ⅱ at 48 h and 4 d after treatment were lower than those of group Ⅰ.And the difference at 48 h after treatment had statistical significance[(18.40 ±0.10)× 109/L vs.(20.60 ± 2.36) × 109/L,(6.12 ± 1.02) kU/L vs.(8.02 ± 1.54) kU/L,(2.14 ±0.21) mg/L vs.(3.34 ± 0.04) mg/L,(1.65 ±0.16) kEU/L w.(2.23 ±0.36) kEU/L] (P ＜ 0.01).While the difference at 4 d after treatment had no statistical significance(P＞ 0.05 ).Body temperature at 48 h after treatment,gastrointestinal decompression capacity,anus exhaust time of group Ⅱ were (37.60 + 3.01 )℃,(320.00 ± 76.14) ml/d,(54.00 ± 0.94) h respectively,and they all were superior to those of group Ⅰ[(38.50 ± 2.21 ) ℃,(500.00 ± 80.32) ml/d,(68.00 ± 1.02) h] (P ＜0.01).Conclusions Continuous intravenous pumping of octreotide can effectively protect the intestinal mucosal barrier function,improve intestinal permeability,reduce the trmslocation of intestinal flora,inhibit the incidence and development of enterogenous endotoxemia.And it provides new evidence to support the clinioal application of octreotide in patients with intestinal endotoxemia.

Oxidative stress and apoptosis in gut barrier dysfunction of severe acute pancreatitis / 中华急诊医学杂志

Objective By means of animal study,investigated the gut barrier function in severe acute pancreatitis ( SAP),and role of inflammatory factors releasing,gut mucosa oxidative stress,cell apoptosis in it.Methods The animal experiment was done in the animal center of first people' s hospital,shanghai jiaotong university.Twenty four BALB/c mice were randomized ( random number) divided into two groups with twelve mice each group.The SAP group,mice received six intraperitoneal injections of cerulein at 1-hour intervals, the dose was 50μg/kg, then given one intraperitoneal injection of 10 mg/kg lipopolysaccharide ( LPS from E.Coli) for the induction of severe acute pancreatitis.The control ( sham operation) group,the mice received intraperitoneal injection of 2 ml normal saline for six times at 1-hour intervals.All the animals of each group were averaged to two batches,4 h and 8h after being operated respectively,to be anesthetized and adopted blood and tissue specimen.Then we observed the pathological change of pancreas and gut,scored it.We measured the blood value of diamine oxidase ( DAO),amylase and tumor necrosis factor-α (TNF-α).We detected content of malondialdehyde (MDA),superoxide dismutase (SOD),glutathione (GSH) and activity of xanthine oxidase (XO) in gut mucosa.We detected the casepase-3 activity and cell apopotosis by means of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) in gut mucosa,and conculated the apopotosis index (AI).Then using the PASW 18.0 software,we analyzed the data by anova and t-test,to make sure if the values were statistically different between the two groups and the mechanism of gut barrier dysfunction in panreatitis.Results At 4 h and 8 h after operation,the SAP-group-mice had significantly higher pancreas pathological score (P ＜0.01 ),blood amylase value ( P ＜ 0.05 ),gut pathological score and blood DAO and TNF-α value ( P ＜0.01 ),compared with the contral-group-mice.The gut mucosa MDA content and XO activity of mice in SAP group were significantly higher than which in control group ( P ＜ 0.01 ). The SAP-group-mice had significantly lower gut mucosa SOD content ( P ＜ 0.01 ) and GSH content ( P ＜ 0.05 ),compared with the contral-group-mice.The gut mucosa cells of mice in SAP group had significantly higher caspase-3 activity and apoptosis index than which in control group ( P ＜ 0.01 ).Conclusions In severe acute pancreatitis,inflammatory factors such as TNF-αwere waterfall-style released,induced gut mucosa suffer from ischemia-reperfusion injury,then serious oxidative stress developed in mucosa and activated caspase-3 pathway,inducing gut mucosa cells apoptose seriously,which was an important mechanism of gut barrier dysfunction.

The protective effects of bifidobacterial adhesin on ischemic reperfusion injury of intestine in rats / 中华内科杂志

ObjectiveTo investigate the protection effect of bifidobacterial adhesin for intestine ischemia/reperfusion (I/R) injury on gut barrier function in rat.MethodsSeventy-two male SD rats were randomly divided into sham operation group (n =24), I/R model group (n =24) and pretreatment group of bifidobacterial adhesin (pretreatment group, n = 24).Six rats were anatomized at 6 h, 1 d, 4 d and 7d after inducing I/R model in each group, respectively.The pathological changes of the terminal ilea and the blood levels of TNFα, IL-6, IL-10, diamine oxidase (DAO), and the activity and content of D-lactic acid were observed.ResultsThe blood levels of TNFα, IL-6, DAO and D-lactic acid in I/R model group were significantly higher than sham operation group at all time points (P ＜0.05) , while the blood level of IL-10 was no significantly change.The activity of IL-6 and DAO in pretreatment group was significantly lower than I/R model group at all time points (P ＜ 0.05), the blood level of TNFαt in pretreatment group was significantly lower than I/R model group at 1 d, the blood level of D-lactic was significantly lower than I/R model group at 4 d and 7 d (P ＜ 0.05). Intestinal pathological damages were obviously milder in pretreatment group than I/R model group at all time points (Chiu's pathological scores: 6 h, 3.22 ±0.22 vs 3.57 ±0.20;1 d,3.77 ±0.13 vs 3.90 ±0.12;4 d,2.93 ±0.23 vs 3.07 ±0.21;7 d,2.10 ±0.30 vs 2.22 ±0.17,all P ＜ 0.05).ConclusionThe pretreatment of bifidobacterial adhesin could protect the intestinal mucosa from I/R injury, and alleviate intestinal ischemic reperfusion injury.

The effects of triggering receptor-1 present on myeloid cellsn on intestinal barrier dysfunction with severe acute pancreatitis / 中华急诊医学杂志

ObjectiveTo investigate the relationship between the expression of triggering receptor1 present on myeloid cells ( TREM-1 ) in intestinal tissue and intestinal barrier dysfunction in severe acute pancreatitis (SAP). MethodsSixty-four male Wistar rats were randomly (random number) divided into sham operation group ( SO group, n = 32) and SAP group ( n = 32 ). The SAP model was established by retrograde injection of 5％ sodium deoxycholate into bile-pancreatic duct. Specimens from blood and intestinal tissue were collected 2, 6, 12 and 48 hours after modeling. The levels of D-lactate, diamine oxidase (DAO) and endotoxin in serum were measured with an modified spectro-photometric method. The expressions of TREM-1, IL-1β and TNF-αt mRNA in terminal ileum were detected by RT-PCR. All data were processed with SPSS version 16. 0 package to make one-way ANOVA and Spearman correlation analysis. ResultsThe serum levels of D-lactate, DAO and endotoxin were significantly increased at all intervals in SAP group compared with SO group ( P ＜ 0. 05 ). The expressions of TREM-1, IL-1β and TNF-α mRNA in terminal ileum of rats in SAP group at all intervals were significantly higher than those in SO group (P ＜ 0. 05 ). The expression of TREM-1 mRNA was positively correlated with expressions of IL-1 β and TNF-α mRNA ( r = 0. 956, P = 0. 044; r = 0. 986, P = 0. 015 ), but correlation was not found between expressions of IL-1β mRNA and TNF-α mRNA ( P = 0. 133 ). ConclusionsThe expression of TREM-1mRNA in intestinal tissue of rats with SAP is elevated, leading to the release of inflammatory cytokines and intestinal mucosal injury, indicating TREM-1 might play an important role in the genesis of intestinal barrier dysfunction in rats with SAP.

Clinical study on bacterial translocation in patients after digestive tract reconstruction / 中国现代普通外科进展

Objective:To investigate gut barrier dysfunction and bacterial translocation (BT) in patients who underwent digestive tract reconstruction and to study the relationship between BT and acute systemic inflammatory state (SIRS). Method： Sixty patients who underwent selective digestive tract reconstruction were observed. Blood were collected before surgery and 1, 3, 5 days after surgery to detect plasma diamine oxidase(DAO) and bacterial DNA. PCR analysis was performed with β-Galactosidase gene of Eschenchia coli and 16SrRNA gene as target gene. The SIRS of all the patients were observed for 10 days. Result：All the PCR results before operation were negative, while there was positive in 14 patients after digestive tract reconstruction. There were 23 patients with SIRS after surgery, and 12 patients PCR result were positive among 23 patients with SIRS. 85.7% of the patients(12/14) with positive PCR result had SIRS, while 23.9% patients (11/46) with negative PCR result had SIRS (P<0.01).The positive PCR rate in SIRS was 52.2% (12/23), which was remarkably higher than that without SIRS(5.4%, 2/37, P<0.01).The levels of plasma DAO in patients with positive PCR result was significantly higher than those of the patients with negative PCR result (P<0.01). The levels of plasma DAO in patients with SIRS was significantly higher than those of patients without SIRS (P<0.01). Conclusion:The gut barrier dysfunction was closely related to BT, and BT was closely related to postoperative SIRS. PCR analysis can be used in early diagnosis of BT, the positive PCR result might be a useful early warning sign of postoperative SIRS.

Relationship between expression of triggering receptor-1 on myeloid cells in intestinal tissue and intestinal barrier dysfunction in severe acute pancreatitis / 世界急诊医学杂志（英文）

BACKGROUND: Triggering receptor expressed on myeloid cells-1 (TREM-1) in the intestine was upregulated and correlated with disease activity in inflammatory bowel diseases. Membrane-bound TREM-1 protein is increased in the pancreas, liver and kidneys of patients with severe acute pancreatitis (SAP), suggesting that TREM-1 may act as an important mediator of inflammation and subsequent extra-pancreatic organ injury. This study aimed to investigate the relationship between the expression of TREM-1 in intestinal tissue and intestinal barrier dysfunction in SAP. METHODS: Sixty-four male Wistar rats were randomly divided into a sham operation group (SO group, n=32) and a SAP group (n=32). A SAP model was established by retrograde injection of 5％sodium deoxycholate into the bile-pancreatic duct. Specimens were taken from blood and intestinal tissue 2, 6, 12, and 48 hours after operation respectively. The levels of D-lactate, diamine oxidase (DAO) and endotoxin in serum were measured using an improved spectro-photometric method. The expression levels of TREM-1, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) mRNA in terminal ileum were detected by real-time reverse transcription-polymerase chain reaction (RT-PCR). Specimens of the distal ileum were taken to determine pathological changes by a validated histology score. RESULTS: The serum levels of D-lactate, DAO and endotoxin were significantly increased in each subgroup of SAP compared with the SO group (P<0.01, P<0.05). The expression levels of TREM-1, IL-1β and TNF-α mRNA in the terminal ileum in each subgroup of SAP were significantly higher than those in the SO group (P<0.01, P<0.05). The expression level of TREM-1mRNA was positively correlated with IL-1β and TNF-α mRNA (r=0.956, P=0.044; r=0.986, P=0.015), but the correlation was not found between IL-1β mRNA and TNF-α mRNA (P=0.133). Compared to the SO group, the pathological changes were aggravated significantly in the SAP group. CONCLUSIONS: The expression level of TREM-1 in intestinal tissue of rats with SAP was elevated, leading to the release of inflammatory mediators and intestinal mucosal injury. This finding indicates that TREM-l might play an important role in the development of intestinal barrier dysfunction in rats with SAP.

Skin Care for Atopic Dermatitis / 대한피부과학회지

BACKGROUND: Interplay of numerous constitutional and precipitating factors are involved in the development of atopic dermatitis (AD). Most of these etiologic factors are not controllable, but barrier dysfunction can be managed. Correction of barrier abnormalities has been accepted as a first-line therapy with effective anti-inflammatory therapy. OBJECTIVE: We wished to establish standardized skin care guidelines for Korean patients who suffer from atopic dermatitis. METHODS: The skin care guidelines were proposed by the Korean Atopic Dermatitis Association (KADA), with a particular emphasis for barrier dysfunction in AD from a review of more than 100 published studies and related documents in the clinical literature by a task force team (TFT) of the KADA. The TFT also evaluated the reliability of the studies based on scientific evidence and the size of the study populations. The TFT disregarded controversial findings and summarized all of the collected studies. RESULTS: Skin care guidelines were proposed by the KADA, particularly for barrier dysfunction in AD. A warm bath for approximately 20 minutes was recommended once daily. Soap can be used two or three times per week and aggressive skin scrubbing should be avoided. A topical moisturizer should be applied at least twice a day. Immediate application of emollient is required after bathing. The donning of clothing made of cotton is recommended. Nails should be cut short to reduce skin damage caused by a skin scratch. CONCLUSION: Application of moisturizer immediately after appropriate bathing may be the most important treatment for skin care. A topical steroid should be used whenever AD develops. Nail care and proper clothing are also be helpful for the prevention of dermatitis aggravation. Proper ways to bathe, use of moisturizer and topical steroids and nail care and appropriate clothing were considered.

Advancement in Intestinal Barrier Dysfunction of Severe Acute Pancreatitis / 中国普外基础与临床杂志

Objective To summarize the recent progress in pathogenetic,diagnostic and therapeutic researches on the intestinal barrier dysfunction(IBD) of severe acute pancreatitis(SAP).Methods The advancement of IBD in SAP,which was published recently at home and abroad,was collected and reviewed.Results The pathogenesis of IBD in patients with SAP was complex.Ischemia-reperfusion injury,endotoxin,inflammatory mediators and gastrointestinal hormone played an important role in the process of IBD.There were many ways to detect IBD,and the ratio of lactulose and mannitol,plasma diamine oxidase were relatively ideal markers.Medical therapies,such as treatment of SAP and maintaining the perfusion of intestines,were essential to cure IBD.On this basis,the propulsives,nutritional support and traditional Chinese drugs should be administered reasonably.Conclusions IBD is a sophisticated process of pathophysiology.In recent years,abundant of animal experiments and clinical researches have provided new clue for prevention and cure of IBD,but further researches are still needed on the mechanism of the cells and molecules implicated.

Effects of vasodilator stimulated phosphoprotein on intestinal barrier dysfunction after hemorrhagic shock in rats / 中华消化外科杂志

Objective To observe the effects of vasedilator stimulated phosphoprotein (VASP) of small intestine mucous membrane on the intestinal barrier dysfunction during hemorrhagic shock (HS) in rats. Methods Forty Wistar rats were divided into normal group, HS 1 hour, HS 2 hours and HS 4 hours groups, as well as HS 2 hours + cyclic adenosine monophosphate (cAMP) treatment group. The activity of sermn diamine oxidase (DAO), content of hematoplasma D-lactic acid of each group were determined, and their relationship with the expression of VASP was analyzed. Results The expression of VASP was increased, serum DAO activity and hematoplasma D-lactic acid content were decreased by cAMP in rats at 2 hours after HS. There were differences upon the levels of VASP expression, DAO activity and hematoplasma D-lactic acid content between HS 2 hours group and HS 2 hours + cAMP treatment group ( t = 18.62, 9.28, 2.83, P < 0.05 ). The serum DAO activity increased while VASP expression decreased significantly after HS, which showed an obvious negative correlation between the two indexes (r=-0.95, P<0.05). Conclusions The decrease of VASP contributes to the intestinal barrier dysfunction after HS in rats, while the intestinal barrier dysfunction can be alleviated by cAMP.