Study on anisodamine hydrobromide improves Th17/Treg imbalance in resuscitated pigs / 中华危重病急救医学

Objective:To investigate whether anisodamine can regulate the ratio of helper T helper cells/regulatory T cells (Th17/Treg) and its protective effect on animals after resuscitation.Methods:Twenty-four Beijing white minipigs were randomly divided into sham operation group (Sham group), resuscitation and normal saline group (SA group), and resuscitation and anisodamine hydrobromide group (AH group), with 8 pigs in each group. In SA group and AH group, ventricular fibrillation was induced by continuous stimulation with intraventricular electrodes for 8 minutes and then resuscitated to establish ischemia/reperfusion (I/R) model. In SA group, after cardiopulmonary resuscitation (CPR), only normal saline was intravenously infused, while in AH group, normal saline and anisodamine hydrobromide were given intravenously at the same time point. Hemodynamic indexes, arterial blood gas analysis indexes, interleukins (IL-17, IL-10) levels in venous blood and IL-17/IL-10 ratio were recorded at 6 different time points: baseline, immediately after return of spontaneous circulation (ROSC), 1 hour, 2 hours, 4 hours and 6 hours after ROSC. The animals were sacrificed at 6 hours after ROSC, and intestinal lymphatic tissues were taken to observe pathological changes under light microscope. At the same time, the levels of IL-17 and IL-10 in intestinal lymphatic tissue were measured (the ratio of IL-17/IL-10 represents the ratio of Th17/Treg cytokines) to evaluate the immune status of the resuscitated animals. The bacterial translocations of different groups were evaluated by culturing intestinal lymphoid tissue.Results:With the extension of ROSC time, the levels of IL-17 in venous blood and the IL-17/IL-10 ratio in pig blood samples continued to decrease, while the levels of IL-10 continued to increase. From 2 hours after ROSC, the IL-17/IL-10 ratio in AH group was significantly higher than that in SA group continued until at 6 hours after ROSC (0.79±0.05 vs. 0.49±0.08, P < 0.05). Light microscopy showed that the number and size of lymph nodules in the cortex of intestinal lymphatic tissue were less in AH group, compared with SA group. Compared with Sham group, the levels of IL-17 and IL-17/IL-10 ratio also decreased in intestinal lymphatic tissue at 6 hours after ROSC [IL-17 (ng/L): 155.23±0.92, 178.76±7.25 vs. 209.21±19.82, IL-17/IL-10 ratio: 1.43±0.13, 1.92±0.18 vs. 3.30±0.31, all P < 0.05], and IL-10 increased significantly (ng/L: 109.85±11.60, 93.55±81.83 vs. 63.45±0.62, all P < 0.05); IL-17/IL-10 ratio in AH group was significantly higher than that in SA group (1.92±0.18 vs. 1.43±0.13, P < 0.05). Tissue culture indicated the intestinal bacterial translocation after resuscitation, suggesting that the animals had immunosuppression and the increased risk of intestinal secondary infection after resuscitation. Compared with SA group, the risk of bacterial translocation was lower than that in AH group [62.5% (5/8) vs. 87.5% (7/8), P < 0.05]. Conclusions:Anisodamine plays an immunomodulatory role by affecting the balance of Th17/Treg cytokines in resuscitated animals, so as to reduce the risk of intestinal secondary infection and has an organ protective effect.

Protection of intestinal immune barrier against ischemia/reperfusion injury in a swine model using anisodamine hydrobromide

Abstract Intestinal ischemia/reperfusion (I/R) causes barrier impairment and bacterial influx. This study explored the protective effects of anisodamine hydrobromide (AH) on intestinal I/R injury caused by cardiopulmonary resuscitation (CPR) after cardiac arrest (CA). After successful CPR, minipigs were randomly divided into two groups (n = 8): saline and AH (4 mg/kg), and then treated with saline or AH via central venous injection, respectively. The same procedures without ventricular fibrillation initiation were conducted in the Sham group (n = 8). Levels of interferon gamma (IFN-γ) and interleukin 4 (IL-4) were measured at different time points (0, 0.5, 1, 2, 4, and 6 h) in serum and 6 h in gut associated lymphoid tissues (GALTs) after the return of spontaneous circulation (ROSC) to evaluate changes in the proportion of T-helper type 1 (Th1) and T-helper type 2 (Th2). Moreover, the positive culture rates of GALTs were examined to evaluate bacterial translocation. AH treatment markedly alleviated aberrant arterial blood gas and hemodynamics as well as intestinal macroscopic and morphological changes after CPR. Moreover, AH treatment significantly increased IFN-γ and decreased IL-4 in both serum and GALTs. Furthermore, AH treatment dramatically decreased positive bacterial growth in GALTs. AH treatment mitigated immunosuppression caused by intestinal I/R and protected the intestinal immune barrier against bacterial translocation, thereby reducing the risk of secondary intestinal infection

Progress on the composition of intestinal mucosal barrier and its relationship with liver injury / 国际儿科学杂志

The complete mucosal barrier of the healthy intestine is the line of defense to prevent the translocation of substances.Many animal models and human pathological studies have proved that the changes of intestinal mucosal barrier function are closely related to the occurrence and treatment of liver disease.This review summarizes the composition of intestinal mucosal barrier, its interaction with liver injury and potential therapeutic targets.

The effects and mechanisms of chronic stress on intestinal and liver injuries in rats / 中华行为医学与脑科学杂志

Objective:To investigate the effects and its mechanism of chronic unpredictable stress on intestine and liver injuries in rats, and explore the possibility of the existence of brain-gut-liver axis.Methods:Twenty male SD rats were randomly divided into control group and stress group (with 10 in each group). The rats in the stress group were stimulated by chronic unpredictable stress for 4 weeks to prepare the chronic stress model. The rats in the control group were fed normally without stress stimulation. After modeling, ten rats in the control group and seven rats in the stress group were included. The depressive behavior of the two groups was evaluated by sugar water preference experiment. Then the rats were sacrificed. The diversity of gut flora in intestinal feces was analyzed by 16S rRNA sequencing analysis. The pathological injuries of ileum and liver were detected by HE staining. The expressions of occludin in ileum and Toll-like receptor 4 (TLR4) in liver were detected by immunohistochemistry. The expression of TLR4 protein in liver tissue was detected by Western blot. The level of lipopolysaccharide (LPS) in rat portal vein serum was detected by AZO chromogenic limulus test and blood biochemical method was used to detect liver function.Statistical analysis was performed using SPSS 25.0 software, and t-test or Mann-Whitney U test was used for comparison between the two groups. Using STAMP software, Wilcoxon rank sum test was used to analyze the difference in bacterial abundance between the two groups. Results:The consumption of sugar water ((7.86±0.90)ml) and the preference rate of sugar water ((43.06±5.65)%) in the stress group were lower than those in the control group ((15.10±1.51)ml, (76.81±6.44)%), and the difference were statistically significant ( t=11.33, 11.16, both P<0.01). Chronic stress caused pathological damage to rat ileum tissue. Compared with the control group, the ileum villi of rats in the chronic stress group were longer ((448.93±12.71)μm, (497.12±16.72)μm, t=-5.88, P<0.01) and thicker ((81.99±16.54)μm, (133.93±6.78)μm, t=-7.12, P<0.01), and the expression of occludin was significantly down-regulated ((0.236±0.011), (0.130±0.026), t=9.12 , P<0.01), the LPS level increased significantly ((18.83±2.62)EU/L, (38.64±2.51)EU/L, t=-5.79, P<0.01). The Beta diversity of rat intestinal flora changed under chronic stress, and the abundance of WPS-2 phylum in intestinal tract of rats in stress group was higher than that in control group ( t=2.76, P<0.05). Chronic stress caused pathological damage to the liver tissue of rats. Compared with the control group, the expression of TLR4 protein in the liver tissue of the chronic stress group increased ((0.169±0.014), (0.475±0.034), Z=-2.37, P<0.05). Compared with the control group, the ALT ((39.7±6.2)U/L, (82.9±43.1)U/L, Z=-2.35, P<0.05) and AST((130.9±28.9)U/L, (472.7±263.3)U/L, Z=-2.64, P<0.05) levels of the chronic stress group increased, especially in AST. Conclusion:Chronic stress cause synchronous damage to the intestine and liver in rats. The mechanism may be related to the results caused by chronic stress such as the changes of the diversity of intestinal flora, the increasing of intestinal permeability, the action of LPS translocated through portal vein blood on TLR4 in liver.

Enterocolitis necrotizante / Necrotizing enterocolitis

Resumen La enterocolitis necrotizante (ECN) es la urgencia más frecuente en el periodo neonatal asociada al sistema digestivo; afectando principalmente a los neonatos pretérmino con muy bajo peso al nacer. La etiología continúa siendo desconocida, se considera una enfermedad multifactorial, donde la prematuridad es el principal factor de riesgo, todo esto relacionado con la inmadurez del tracto gastrointestinal, una motilidad disminuida que genera una mayor permeabilidad en la mucosa y con esto facilita la translocación bacteriana. Su diagnóstico suele ser muy complejo y suele pasar desapercibido en muchas ocasiones generando una mortalidad importante de hasta el 30% donde su principal complicación es la perforación intestinal y el consiguiente shock séptico. Las opciones terapéuticas se dividen en 2 grupos: médico y quirúrgico, ambos con complicaciones importantes que afectan el desarrollo de los niños que la padecen donde las más importantes abarcan desde alteraciones del crecimiento y neurodesarrollo hasta síndrome de intestino corto y desnutrición.

Abstract Necrotizing enterocolitis (NEC) is the most common emergency in the neonatal period associated with the digestive system; mainly affecting preterm neonates with very low birth weight. Etiology remains unknown, considered a multifactorial disease, all this related to the immaturity of the gastrointestinal tract, a decreased motility that generates greater permeability in the mucosa and with this facilitates bacterial translocation. Diagnosis is usually very complex and often goes unnoticed on many occasions leading to a significant mortality of up to 30% where its main complication is intestinal perforation and consequent septic shock. Therapeutic options are divided into 2 groups: medical and surgical, both with major complications affecting the development of children with it where the most important from growth and neurodevelopmental alterations to short bowel syndrome and malnutrition.

Vitamin D Improves Intestinal Barrier Function in Cirrhosis Rats by Upregulating Heme Oxygenase-1 Expression

Intestinal barrier dysfunction always accompanies cirrhosis in patients with advanced liver disease and is an important contributor facilitating bacterial translocation (BT), which has been involved in the pathogenesis of cirrhosis and its complications. Several studies have demonstrated the protective effect of Vitamin D on intestinal barrier function. However, severe cholestasis leads to vitamin D depletion. This study was designed to test whether vitamin D therapy improves intestinal dysfunction in cirrhosis. Rats were subcutaneously injected with 50% sterile CCl₄ (a mixture of pure CCl₄ and olive oil, 0.3 mL/100 g) twice a week for 6 weeks. Next, 1,25(OH)₂D₃(0.5 µg/100 g) and the vehicle were administered simultaneously with CCl₄ to compare the extent of intestinal histologic damage, tight junction protein expression, intestinal barrier function, BT, intestinal proliferation, apoptosis, and enterocyte turnover. Intestinal heme oxygenase-1 (HO-1) expression and oxidative stress were also assessed. We found that vitamin D could maintain intestinal epithelial proliferation and turnover, inhibit intestinal epithelial apoptosis, alleviate structural damage, and prevent BT and intestinal barrier dysfunction. These were achieved partly through restoration of HO-1 and inhibition of oxidative stress. Taken together, our results suggest that vitamin D ameliorated intestinal epithelial turnover and improved the integrity and function of intestinal barrier in CCl₄-induced liver cirrhotic rats. HO-1 signaling activation was involved in these above beneficial effects.

Effects of vitamin D3 on intestinal mucosal barrier of mice with severe burns / 中华烧伤杂志

Objective@#To explore the effects of vitamin D3 on intestinal mucosal barrier of mice with severe burns.@*Methods@#Forty-two C57BL/6C male mice aged eight to twelve weeks were divided into vitamin D3 vehicle+ sham injury group of seven mice, vitamin D3 vehicle+ burn injury group of fourteen mice, vitamin D3+ sham injury group of seven mice, and vitamin D3+ burn injury group of fourteen mice according to random number table. Mice in vitamin D3 vehicle+ sham injury group and vitamin D3 vehicle+ burn injury group were injected with vehicle of vitamin D3 at a dose of 0.1 mL intraperitoneally at 1, 24, and 48 h before burn experiment. Mice in vitamin D3+ sham injury group and vitamin D3+ burn injury group were injected with vitamin D3 at a dose of 100 ng/kg dissolved in 0.1 mL vehicle intraperitoneally at the same time points. Mice in vitamin D3 vehicle+ burn injury group and vitamin D3+ burn injury group were inflicted with 30% total body surface area full-thickness dermal scald (hereinafter referred to as burn) on the back by 98 ℃ hot water for 3 to 4 seconds. And mice in vitamin D3 vehicle+ sham injury group and vitamin D3+ sham injury were treated with 37 ℃ water on the back for 3 to 4 seconds to simulate injury. Seven mice in vitamin D3 vehicle+ sham injury group and seven mice in vitamin D3+ sham injury group at post injury hour (PIH) 24, and seven mice in vitamin D3 vehicle+ burn injury group and seven mice in vitamin D3+ burn injury group at PIH 6 and 24 were sacrificed respectively to collect mesentery lymph nodes, spleens, livers, and intestinal tissue. The mesentery lymph nodes, spleens, and livers of mice in each group were collected to observe growth of bacteria, and number of bacteria was counted. Intestinal tissue of mice in each group was collected to detect protein expressions of zonal occludin 1 (ZO-1) and occludin by immunohistochemistry staining method, distribution of ZO-1 by immunofluorescence staining method, and expression of occludin by Western blotting. Data were processed with Kruskal-Wallis H test, Nemenyi test, one-way analysis of variance, t test, and Bonferroni correction.@*Results@#(1) At PIH 6 and 24, bacterial counts of mesentery lymph nodes, livers, and spleens of mice in vitamin D3 vehicle+ burn injury group were significantly higher than those of mice in vitamin D3 vehicle+ sham injury group (P<0.05). At PIH 6, bacterial counts of livers and spleens of mice in vitamin D3+ burn injury group were significantly lower than those of mice in vitamin D3 vehicle+ burn injury group (P<0.05). At PIH 24, bacterial counts of mesentery lymph nodes and livers of mice in vitamin D3+ burn injury group were significantly lower than those of mice in vitamin D3 vehicle+ burn injury group (P<0.05). (2) At PIH 6 and 24, expressions of ZO-1 and occludin of intestinal tissue of mice in vitamin D3 vehicle+ burn injury group were significantly lower than those of mice in vitamin D3 vehicle+ sham injury group, and expressions of ZO-1 and occludin of intestinal tissue of mice in vitamin D3+ burn injury group were close to those of mice in vitamin D3+ sham injury group. At PIH 6 and 24, expressions of ZO-1 and occludin of intestinal tissue of mice in vitamin D3+ burn injury group were significantly higher than those of mice in vitamin D3 vehicle+ burn injury group. (3) At PIH 6 and 24, compared with that of mice in vitamin D3 vehicle+ sham injury group, distribution of ZO-1 of intestinal mucosal epithelium of mice in vitamin D3 vehicle+ burn injury group was discontinuous. Distribution of ZO-1 of intestinal mucosal epithelium of mice in vitamin D3+ sham injury group was normal, and the distribution of ZO-1 of intestinal mucosal epithelium of mice in vitamin D3+ burn injury group was with good continuity. (4) At PIH 6 and 24, expressions of occludin of intestinal tissue of mice in vitamin D3 vehicle+ burn injury group were 0.720±0.003, 0.638±0.052 respectively, significantly lower than 0.918±0.003 of mice in vitamin D3 vehicle+ sham injury group (t=57.33, 5.36, P<0.05). At PIH 6 and 24, expressions of occludin of intestinal tissue of mice in vitamin D3+ burn injury group were 0.994±0.058, 1.064±0.060, close to 0.938±0.023 of mice in vitamin D3+ sham injury group (t=0.91, 1.96, P>0.05). At PIH 6 and 24, expressions of occludin of intestinal tissue of mice in vitamin D3 vehicle+ burn injury group were significantly lower than those of mice in vitamin D3+ burn injury group (t=4.75, 5.35, P<0.05).@*Conclusions@#Intestinal bacterial translocation can occur in the early stage of severe burn. And vitamin D3 plays a protective role in the intestinal mucosal barrier post severe burn to reduce the bacterial translocation by protecting tight junction proteins of intestinal epithelium.

Effect of broad-spectrum antibiotics on bacterial translocation in burned or septic rats / 中华医学杂志(英文版)

Background:@#Antibiotics are frequently used to treat critically ill patients, and its use is often accompanied by intestinal dysbiosis that might further lead to bacterial translocation (BT). Nevertheless, studies on the relationship between antibiotic therapy and BT are rare. In the present study, we investigated the effect of broad-spectrum antibiotics on BT in an experimental rat model of burn or sepsis injury.@*Methods:@#The septic rat model was simulated by a second insult with lipopolysaccharides after burn injury. Ninety-two male Sprague-Dawley rats were randomly divided into control, burn, and sepsis groups (n = 8 or 9, each group), and the latter two groups were then treated with imipenem or ceftriaxone for 3 or 9 days. The mesenteric lymph nodes, liver, lungs, and blood were collected at each time point under sterile conditions for quantitative bacterial culture and strain identification. The differences between the groups were compared by Fisher exact test or Mann-Whitney U test.@*Results:@#Only minimal Escherichia coli translocation to the mesenteric lymph nodes was observed in the normal control group, in which the BT rate was 12.5%. Burn injury did not affect the BT rate (Burn group vs. Control group, 12.5% vs. 12.5%, P = 1.000), whereas the BT rate showed an increased trend after the second insult with lipopolysaccharide (Sepsis group vs. Control group, 44.4% vs. 12.5%, P = 0.294), and many strains of Enterobacteria spp. were detected in distant organs (liver, lung, and blood) [Sepsis group vs. Control group, 0 (0,3) vs. 0 (0,0), U = 20, P = 0.045]. After the antibiotic treatment, BT to the distant organs was increased in burned rats [Burn IT3 group vs. Burn group, 0 (0,2) vs. 0 (0,0); Burn IT9 group vs. Burn group, 0 (0,1) vs. 0 (0,0); Burn CT9 group vs. Burn group, 0 (0,2) vs. 0 (0,0); all U = 20 and P = 0.076] but decreased in septic rats [Sepsis CT3 group vs. Sepsis group, 0 (0,0) vs. 0 (0,3), U = 20, P = 0.045]. The total amount of translocated bacteria, regardless of which antibiotic was used, was increased in burned rats [Burn IT9 group vs. Burn group, 2.389 (0,2.845) vs. 0 (0,2.301) Log10 colony-forming units (CFU)/g, U = 14, P = 0.034; Burn CT3 group vs. Burn group, 2.602 (0,3.633) vs. 0 (0,2.301) Log10 CFU/g, U = 10.5, P = 0.009], but there was a slightly decreased trend in septic rats [Sepsis IT9 group vs. Sepsis group, 2.301 (2,3.146) vs. 0 (0,4.185) Log10 CFU/g, U = 36, P = 0.721; Sepsis CT9 group vs. Sepsis group, 2 (0,3.279) vs. 0 (0,4.185) Log10 CFU/g, U = 32.5, P = 0.760]. Remarkably, the quantity of Enterococci spp. dramatically increased after broad-spectrum antibiotic treatment in both the burned and septic groups [Burn IT3 group vs. Burn group, 1 (0,5.164) vs. 0 (0,0) Log10 CFU/g, U = 16; Burn IT9 group vs. Burn group, 1 (0,2.845) vs. 0 (0,0) Log10 CFU/g, U = 16; Burn CT3 group vs. Burn group, 2.602 (0,3.633) vs. 0 (0,0) Log10 CFU/g, U = 8; Burn CT9 group vs. Burn group, 1 (0,4.326) vs. 0 (0,0) Log10 CFU/g, U = 16; Sepsis IT3 group vs. Sepsis group, 2.477 (0,2.903) vs. 0 (0,0) Log10 CFU/g, U = 4.5; Sepsis IT9 group vs. Sepsis group, 2 (0,3.146) vs. 0 (0,0) Log10 CFU/g, U = 9; Sepsis CT3 group vs. Sepsis group, 1.151 (0,2.477) vs. 0 (0,0) Log10 CFU/g, U = 18; Sepsis CT9 group vs. Sepsis group, 2 (0,3) vs. 0 (0,0) Log10 CFU/g, U = 13.5; all P < 0.05].@*Conclusions:@#Broad-spectrum antibiotics promote BT in burned rats but prevent BT in septic rats, especially preventing BT to distant organs, such as the liver and lung. Moreover, Enterococci spp. with high drug resistance and high pathogenicity translocated most after antibiotic treatment.

Effects of Endotoxin Affinity Adsorbent SPV on Intestinal Permeability and Bacterial Translocation in Hemorrhagic Shock Model Rats / 中国药房

OBJECTIVE： To observe the effects of endotoxin affinity adsorbent SPV on intestinal permeability and bacterial translocation in hemorrhagic shock model rats.  METHODS： Totally 85 male SD rats were randomly divided into normal group （5 rats）， shock group （each 5 rats at each time point， 20 rats in total）， SPV low-dose， medium-dose and high-dose groups （Montmorillonite powder 0.3 g， Polymyxin B sulfate 0.5 mg， Vitamin B6 5 mg dissolved in normal saline to obtain SPV solution 5 mL， as low dose； medium and high dose were 2 or 3 times as high as low dose. Each 5 rats of each group at each time point， 60 rats in total）. Administration groups were given SPV solution intragastrically 5， 10， 15 mL once， respectively； normal group and shock group were given normal saline 5 mL intragastrically once. Thirty minutes after last medication， other groups received femoral artery catheterization and bleeding to induce hemorrhagic shock model， except for normal group. The activities or contents of diamine oxidase （DAO）， endotoxin and D-lactic acid， positive rates of intestinal bacterial translocation were detected in each group at 1， 4， 8， 16 h after recovery. RESULTS： Compared with normal group， the activities of DAO of rats in shock group were enhanced significantly， and the serum contents of endotoxin and D-lactic acid were increased significantly （P＜0.05）. Compared with shock group， the activities of DAO were decreased significantly in SPV groups （at each time point during 1-16 h）； the serum contents of endotoxin and D-lactic acid （at each time point during 1-16 h）， positive rates of intestinal bacterial translocation （SPV low-dose group at each time point during 4-16 h， SPV medium-dose and high-dose groups at each time point during 1-16 h） were decreased significantly （P＜0.05）. Above indexes in SPV medium-dose and high-dose groups （at each time point during 1-16 h） were significantly lower than those of SPV low-dose group （P＜0.05）. There was no statistical significance in above indexes between SPV medium-dose group and high-dose group （P＞0.05）. CONCLUSIONS： The endotoxin affinity adsorbent SPV can improve the permeability of the intestinal wall and inhibit bacterial translocation in hemorrhagic shock model rats in dose-dependent manner. The effects of which may be associated with reducing the activities or contents of serum DAO， endotoxin， D-lactic acid， and down-regulating the positive rate of bacterial translocation.

Intestinal barrier and intestinal endotoxemia / 中国小儿急救医学

Intestinal dysfunction can be induced under infection,trauma or stress. The intestinal mu＿cosal barrier injury is a common pathophysiological process in critically ill patients. The bowel is one of the earliest and most severely affected organ under severe infection and tissue hypoxia＿ischemia. Intestinal dam＿age leads to a large number of bacterial flora shift,resulting in intestinal infection,intestinal endotoxemia and intestinal sepsis. In recent years, gastrointestinal barrier function has attracted people′s attention, which is closely related to the prognosis of critically ill patients. This article described the normal intestinal barrier function,the effects of trauma and stress on intestinal barrier function, bacterial group migration, and the effects of endotoxin translocation on the body and its prevention and treatment.

Association between intestinal barrier and bacterial translocation in patients with liver cirrhosis / 临床肝胆病杂志

Decompensated liver cirrhosis has various life-threatening complications such as spontaneous bacterial peritonitis, hepatic encephalopathy, and sepsis, and the development of such complications is closely associated with bacterial translocation. Pathological bacterial translocation in patients with liver cirrhosis is caused by the impairment of intestinal barrier function, and thus bacteria can cross the intestinal barrier and enter the mesenteric lymph nodes or other sites. Therefore, a better understanding of the association between intestinal barrier and bacterial translocation in liver cirrhosis can provide new theoretical support for the treatment of liver cirrhosis. This article discusses the four major components of the intestinal barrier, namely the mechanical barrier, the immune barrier, the chemical barrier, and the biological barrier, as well as their changes during bacterial translocation.

Influence of intestinal dysbacteriosis on liver cirrhosis / 临床肝胆病杂志

Liver cirrhosis is a progressive chronic liver disease due to one or more causes, with diffuse fibrosis in liver tissue, pseudolobules, and regenerative nodules as major histological features. Once liver cirrhosis enters the decompensated stage, the liver and several other organs are injured, which can hardly be recovered or reversed. This article introduces the process of Toll-like receptors in recognizing the changes in intestinal flora and the influence of this process on the development and progression of liver cirrhosis, as well as the protective effect of bile acid against liver cirrhosis by regulating intestinal flora. This article also reviews the advances in delaying liver cirrhosis after the treatment of intestinal dysbacteriosis and points out that the treatment of intestinal dysbacteriosis may become the major direction of liver cirrhosis treatment in future.

Intestinal barrier and intestinal endotoxemia / 中国小儿急救医学

Intestinal dysfunction can be induced under infection,trauma or stress. The intestinal mu-cosal barrier injury is a common pathophysiological process in critically ill patients. The bowel is one of the earliest and most severely affected organ under severe infection and tissue hypoxia-ischemia. Intestinal dam-age leads to a large number of bacterial flora shift,resulting in intestinal infection,intestinal endotoxemia and intestinal sepsis. In recent years,gastrointestinal barrier function has attracted people′ s attention,which is closely related to the prognosis of critically ill patients. This article described the normal intestinal barrier function,the effects of trauma and stress on intestinal barrier function,bacterial group migration,and the effects of endotoxin translocation on the body and its prevention and treatment.

Intestinal mucosal barrier damage and the development of cachexia in colon cancer patients / 中华普通外科杂志

Objective To investigate the association of bacterial translocation (BT) with cachexia in colonic cancer patients.Methods From September 2015 to May 2017 the clinical data of 292 colon cancer patients were studied at Qingdao Municipal Hospital.The bacteria in peripheral blood and mesenteric lymph nodes were detected by bacterial culture,and the bacterial DNA in peripheral blood was detected by PCR technique to determine the occurrence of bacterial translocation.Intestinal epithelial T-cell subsets and NK cells were evaluated using flow cytometry.Western blot and immunofluorescence were used to check tight junction proteins Occludin,Claudins-2,Zonula occluden-2 in intestinal epithelium.Fluorescence in situ hybridization and immunohistochemistry were used to detect the translocated bacteria and endotoxin.Results Compared with noncachectic patients,cachectic patients had a significandy higher BT ratio (27.8％ vs.7.2％,x2 =20.871,P ＜ 0.001).BT in the intestinal mucus layer was associated with lower levels of T-cell subsets and NK cells in the intestinal epithelium in BT(+) patients (CD3 + T:36.69％ ±5.87％ vs.41.63％ ±5.03％,CD4+T:44.08％ ±5.12％ vs.49.58％ ±7.01％,CD8+T:65.68％ ±5.51％ vs.61.43％ ± 5.58％,CD4+ T/CD8+ T:0.71％ ± 0.21％ vs.0.91％ ±0.23％,NK:27.86％ ± 3.93％ vs.34.69％ ± 4.52％,all P ＜ 0.01).Endotoxin was detected within the small intestinal wall in BT(+) patients and claudin-2 expression increased (0.63 ± 0.13 vs.0.21-± 0.06,t =-2.936,P ＜ 0.01),whereas Occludin and Zonula occluden-2 expressions decreased (0.37 ± 0.13 vs.0.84±0.17,0.16±0.07 vs.0.58±0.19,t=2.151,2.111,bothP＜0.05).Conclusions BTmay contribute to the development of colon cancer cacheria,and tight junction could be the gateway of BT.

16S rDNA-based molecular profiles of circulating bacteria in patients with severe acute pancreatitis / 肠外与肠内营养

Objective:This study aims to characterize the bacterial profile presenting in peripheral blood of severe acute pancreatitis (SAP) patients and investigate the potential role of circulating microorganisms in the development of systemic infection.Methods:A total of 30 patients with SAP were recruited in this study and divided into three groups:infected,sepsis and Septic shock (n =10 for each group).The peripheral blood was collected sterilely for extraction of DNA,which was subsequently amplified using the universe primers targeted the V6-V8 region of 16S ribosomal RNA genes.The amplicons were separated by denaturing gradient gel electrophoresis (DGGE),and then the gels were stained and photographed.The bands were cut out and sequenced to determine the closest bacterial relatives.Results:As shown in DGGE profile,multiple DNA bands (3 to 8 bands) were detected in peripheral blood from all (100％) of SAP patients complicated with septic shock.The microorganisms most frequently presenting in the blood of these cases included Escherichia coli,Bacillus coagulans,Pseudomonas putida,Pseudomonas aeruginosa,and Klebsiella pneumonia,with an incidence of 40 ％ or higher.In patients with sepsis,bacterial DNA consisting of 2 to 4 bands was observed in 90％ of the blood samples.The most common bacterial species was Pseudomonas putida (60％),followed by Shigella flexneri (40％),Staphylococcus aureus (30％) and Enterococcusfaecium (20％).By contrast,the positive rate of blood bacterial DNA was relatively lower in infected patients (70 ％).Of them,single bacterial species was commonly found in the blood samples.Conclusions:Our data showed that the bacterial profiles presenting in peripheral blood are distinct among SAP patients with different manifestations.Polymicrobial translocation could contribute to the development of systemic infection,offering novel insights into the pathogenesis of sepsis in SAE The findings are helpful for the prevention and treatment for bacterial infection and complications of SAP.

Reduction of venous pressure during the resection of liver metastases compromises enteric blood flow: IGFBP-1 as a novel biomarker of intestinal barrier injury

OBJECTIVES: Disruption of the intestinal barrier and bacterial translocation commonly occur when intestinal blood flow is compromised. The aim of this study was to determine whether liver resection induces intestinal damage. METHODS: We investigated intestinal fatty-acid binding protein and insulin-like growth factor binding protein levels in the plasma of patients who underwent liver resection. RESULTS: We show that liver resection is associated with significant intestinal barrier injury, even if the Pringle maneuver is not performed. CONCLUSION: We propose the use of insulin-like growth factor binding protein-1 as a novel biomarker of intestinal damage in such situations.

Bacterial translocation and mortality on rat model of intestinal ischemia and obstruction

Abstract Purpose: To develop an experimental model of intestinal ischemia and obstruction followed by surgical resection of the damaged segment and reestablishment of intestinal transit, looking at bacterial translocation and survival. Methods: After anesthesia, Wistar rats was subject to laparotomy, intestinal ischemia and obstruction through an ileal ligature 1.5cm of ileum cecal valve; and the mesenteric vessels that irrigate upstream of the obstruction site to approximately 7 to 10 cm were ligated. Abdominal wall was closed. Three, six or twenty-four hours after, rats were subject to enterectomy followed by an end to end anastomosis. After 24h, mesenteric lymph nodes, liver, spleen and lung tissues were surgically removed. It was studied survival rate and bacterial translocation. GraphPadPrism statistical program was used. Results: Animals with intestinal ischemia and obstruction for 3 hours survived 24 hours after enterectomy; 6hx24h: survival was 70% at 24 hours; 24hx24h: survival was 70% and 40%, before and after enterectomy, respectively. Culture of tissues showed positivity on the 6hx24h and negativity on the 3hx24h. Conclusion: The model that best approached the clinic was the one of 6x24h of ischemia and intestinal obstruction, in which it was observed bacterial translocation and low mortality rate.

Correlation between acute gastrointestinal injury severity and intestinal microenvironment in rabbits with severe multiple trauma / 中华创伤杂志

Objective To investigate the correlation between acute gastrointestinal injury severity and intestinal microenvironment in rabbits with severe multiple trauma.Methods A total of 60 New Zealand white rabbits were enrolled and randomly assigned into the experimental group (48 rabbits) and control group (12 rabbits).In experimental group,the models of traffic-induced injuries were successful made by using self-made small gravity traction colliders.There were nine rabbits with craniocerebral injury combined with damage of liver and spleen,three with four extremity fractures combined with damage of liver and spleen,12 with rib fractures combined with damage of lungs and pleural effusion,11 with epidural hematoma,contusion and laceration of brain as well as fractures of four extremities and pelvis,and nine with multiple fractures.The injuries had met the criteria of multiple severe traumas according to the injury severity score (ISS).The control group had similar condition with experimental group except for participation in injury model.The parameters at time points of 6,12,24 and 48 h after injury were observed.The parameters were:(1)Enzyme linked immunosorbent assay (ELISA) and high pressure liquid chromatography (HPLC) were used to detect the levels of diamine oxidase (DAO) and the ratio of lactulose to mannitol in urine in order to evaluate the permeability of intestinal mucosal barrier.(2) The small intestinal propulsive rate was detected by carbon pushing mcthod.The interstitial cells of Cajal in the snall intestine wall were observed by transmission electron microscopy.The expression of C-kit was detected by Western blot.By these means,the dynamic function of intestinal mucosal barrier was evaluated.(3) The damage degree of intestinal nucosal barrier was evaluated by pathological observation and Chiu score in the end of small intestine and colon.(4)The intestinal bacterial translocation was evaluated by intestinal microflora culture,mesenteric lymph nodes checking as well as translocation examination of liver and spleen.Results (1) The level of DAO plasma and urine lactulose/mannitol ratio at 6 h after injury increased to varied degrees,and reached the peak during 12-24 h,the correlation analysis of which showed that plasma DAO levels were positively correlated with AGI grade at 6,12 and 24 h (r =0.486 3,0.493 3,0.477 6,P ＜ 0.05).The ratio of urinary lactulose/ mannitol excretion was only positively correlated with AGI grade at 6 and 12 h (r =0.478 5,0.497 2,P ＜ 0.05).(2) The gastrointestinal transmission rate in the experimental group was (48.2 ± 5.2) ％,and that of the control group was (60.3 ± 3.0) ％.The gastrointestinal transmission rate of model group rabbits was significantly longer than that of control group (P ＜ 0.05).(3) The interstitial cells of Cajal decreased dramatically during 12-24 h after the injury,and the expression level of C-kit protein decreased consistently with the increase of AGI grade.(4) The intestinal and colonic mucosa tissues of the experimental group showed obvious necrosis at 12-24 h after injury,and the Chiu score increased gradually with the increase of AGI grade under light microscope.(5) The Enterobacter and Enterococcus in the experimental group increased significantly compared with that in the control group.But the numbers of Bifidobacterium,Lactobacillus,Lactobacillus,Bacteroides as well as the ratio of Bifidobacterium to Enterobacteriaceae significantly decreased(P ＜0.05).(6)The organ bacterial translocation rates were 24％,42％ and 62％ after injury in experimental group (P ＜0.05).Conclusions Acute gastrointestinal injury may occur early after severe multiple trauma,and the injury severity is closely related to the change of intestinal barrier function and bacterial translocation.Early attention and active correction of the change of intestinal microenvironment are of great importance for treatment of multiple trauma.

Research of Acute Necrotizing Pancreatitis Associated with Diabetes Mellitus on Early Bacterial Translocation in Rat / 中山大学学报(医学科学版)

[Objective] To explore whether diabetes mellitus (DM) can influence the early bacterial translocation (BT) and progression of acute necrotizing pancreatitis (ANP) for guiding the early clinical treatment.[Methods] 35 Wistar male rats were randomly allocated to 4 groups,Group ANP associatcd with DM (DM+ANP,n =10):DM underwent induction of ANP;Group DM (n =10):DM underwent laparotomy with only manipulation of the pancreas and duodenum;Group ANP (n =10):non-DM underwent induction of ANP;Group sham operation (SO,n =5):non-DM underwent SO.After 12 h of the induction of ANP or laparotomy,the following parameters were analyzed:bacterial culture and identification of portal vein blood,mesenteric lymph nodes (MLNs),pancreas and liver,and calculate the total incidence of BT;serum amylase and endotoxin levels of portal vein blood;histological assessment of pancreas and ileum lesions.[Results] All animals except 3 in group DM+ANP (mortality rates:30％) and 1 in group ANP (mortality rates:10％) survived the experiment.The total incidence of BT was 23/28 (82.1％) in group DM+ANP whereas 16/36 (44.4％) in group ANP (P =0.002).Gram-positive bacteria were 17/23 (73.9％),3/16 (18.8％) in group DM+ANP and group ANP,respectively (P =0.001).Amylase activity (2302 ± 346) U/L in group ANP increased significantly (P =0.000) compared with other groups.However,group DM+ANP (501 ± 142) U/L decreased significantly (P =0.001) in comparison to group SO.Regarding to endotoxin concentrations and the severity of pancreas and ileum lesions,group DM + ANP increased significantly compared with group ANP,group DM and group SO (P ＜ 0.05).[Conclusion] Gram-positive bacteria translocates more frequently than Gram-negative bacteria in the early period of DM+ANP rats.DM aggravates the progression of ANP and increases early bacterial translocation,endotoxemia and severity of pancreas and ileum lesions.

Gut-origin sepsis / 中国小儿急救医学

Gut-origin sepsis is paid more attention recently.Gut has become an important origin of sepsis for its full of bacteria and toxin and the role in immune regulation,which is hidden in infants and young children.We rarely diagnose gut-origin sepsis deliberately in clinic,but just as a source of infection.In this article,we discussed the pathogenesis including some debated problems and progress of treatment by literature review,hope to help clinicians improve the ability to diagnose and treat gut-origin sepsis.