D-lactic Acid Levels Research Articles

Probiotics ameliorate renal ischemia-reperfusion injury by modulating the phenotype of macrophages through the IL-10/GSK-3β/PTEN signaling pathway.

After renal ischemic reperfusion injury, a series of pathological changes, such as impaired intestinal barrier function, intestinal flora, and endotoxin translocation, are caused by intestinal ischemia and hypoxia, which then trigger systemic inflammatory responses and affect the condition and prognosis of the patients. In this study, a rat model of ischemia-reperfusion injury was established by examining changes in renal function, intestinal barrier function, inflammatory index, oxidative stress, and macrophage phenotypes to evaluate the effect of probiotic VSL#3 on renal ischemia-reperfusion injury. The results showed that, after VSL#3 intervention, the levels of BUN, Scr, Cys C, PRO, and NGAL were all significantly decreased compared with the I/R group, while the value of Ccr showed a significant increase. In addition, the concentrations of MPO, IL-1β, TNF-α, IL-6, ED-1, and PCNA were all significantly lower than those in the I/R group, while the levels of endotoxin, DOA, and D-lactic acid were significantly decreased. Furthermore, the proteins associated with intestinal barrier functions, such as ZO-1, Occludin, and Claudin-1, were significantly upregulated compared with the I/R group. Overall, the VSL#3 intervention group was able to maintain the required number of beneficial intestinal flora and to inhibit the proliferation of harmful bacteria. At the same time, the VSL#3 intervention could also prevent the decrease in the levels of CAT, GSH-PX, H2O2, and T-SOD, while downregulating the expression of Keap1 and Nrf2. After the intervention with the VSL#3, the expression levels of CD68 and CD86 proteins were significantly decreased, while the expression levels of CD163 and CD206 proteins were significantly higher. Further experiments confirmed that the expression of iNOS protein was significantly decreased after the VSL#3 intervention, and the expression of Arg-1 and Ym1 proteins was significantly increased. The VSL#3 was able to induce high expressions of p-GSK-3β and p-PTEN proteins, while the use of IL-10 antibody impaired the effect of the VSL#3. In summary, this research confirms that probiotics can alleviate renal dysfunction caused by ischemia and reperfusion by protecting the intestinal barrier function and maintaining the functions of intestinal flora. The pathway screening test of this study suggests that IL-10/GSK3β/PTEN may play an important role in the process of the prototypic VSL#3 inducing M2 transformation of macrophages.

Clinical study of Rhubarb water extract gavage combined with external application of mirabilite and western medicine in the treatment of severe acute pancreatitis

Objective To investigate the effects of Rhubarb water extract gavage combined with external application of mirabilite and western medicine in the treatment of severe acute pancreatitis (SAP). Methods A total of 62 patients with SAP meeting the inclusion criteria were randomly divided into 2 groups, with 31 cases in each group. The control group was treated with western medicine routine treatment. The study group was given the combination of raw and rhubarb water infusion and mirabilite external application on the basis of the control group. After treated with 28 d, the duration of blood purification, abdominal pain disappeared time, blood amylase recovery time, bowel peristalsis recovery time, hospitalization time and 28 d mortality were observed in the 2 groups. Results The duration of blood purification time, abdominal pain disappeared time, blood amylase recovery time, bowel peristalsis recovery time, hospitalization time of the study group were significantly lower than those of the control group (t=2.434, 2.507, 2.804, 2.318, 2.614, P<0.05). After treatment 1, 3, 7 d, the level of D-lactic acid in the study group was respectively which were significantly lower than those of the control group (t=38.752, 36.975, 36.018, P<0.05). The level of DAO in the study group was respectively which were significantly lower than those of the control group (t=4.119, 4.632, 3.907, P<0.05). The level of ET in the study group was respectively which were significantly lower than those of the control group (t=25.699, 23.805, 24.472, P<0.05). The level of hs-CRP in the study group was respectively which were significantly lower than those of the control group (t=4.791, 5.025, 5.288, P<0.05). The level of PCT in the study group was respectively which were significantly lower than those of the control group (t=4.350, 4.742, 6.012, P<0.05). The APACHE II scores in the study group were respectively which were significantly lower than those of the control group (t=3.988, 4.350, 5.722, P<0.01). Conclusions The application of Rhubarb water extract gavage combined with external application of mirabilite and western medicine in the treatment of severe acute pancreatitis can protect the intestinal mucosal barrier of SAP patients, reduce the level of inflammatory reaction, shorten the time of symptom relief and improve the curative effect. Key words: Severe acute pancreatitis; Rheum officinale; Natrii sulfas; Hypersensitive C reactive protein; procalcitonin

Protective role of down-regulated microRNA-31 on intestinal barrier dysfunction through inhibition of NF-\u03baB/HIF-1\u03b1 pathway by binding to HMOX1 in rats with sepsis

BackgroundIntestinal barrier dysfunction is a significant clinical problem, commonly developing in a variety of acute or chronic pathological conditions. Herein, we evaluate the effect of microRNA-31 (miR-31) on intestinal barrier dysfunction through NF-κB/HIF-1α pathway by targeting HMOX1 in rats with sepsis.MethodsMale Sprague-Dawley rats were collected and divided into the sham group, and the cecum ligation and perforation group which was subdivided after CACO-2 cell transfection of different mimic, inhibitor, or siRNA. Levels of serum D-lactic acid, diamine oxidase and fluorescence isothiocyanate dextran, FITC-DX concentration, and bacterial translocation were detected. Superoxidedismutase (SOD) activity and malondialdehyde (MDA) content were evaluated using the colorimetric method and an automatic microplate reader, respectively. Additionally, the levels of tumor necrosis factor, interleukin (IL)-6, and IL-10 were tested using enzyme-linked immunosorbent assay. The expression of miR-31, HMOX1, NF-κB, HIF-1α, IκB, ZO-1 and Occludin were assessed by reverse transcription quantitative polymerase chain reaction and Western blot analysis.ResultsInhibition of miR-31 decreased intestinal mucosal permeability and intestinal barrier function. The increased levels of miR-31 could cause oxidative damage and affect the expression of inflammatory factors in intestinal tissue of rats. HMOX1 was confirmed as a target gene of miR-31. MiR-31 affected intestinal mucosal permeability and intestinal barrier function, as well as oxidative damage and inflammation level by regulating HMOX1. Down-regulation of miR-31 inhibited NF-κB/HIF-1α pathway related genes by regulating HMOX1 expression. Furthermore, inhibition of miR-31 increased survival rates of rats.ConclusionOverall, the current study found that inhibition of miR-31 protects against intestinal barrier dysfunction through suppression of the NF-κB/HIF-1α pathway by targeting HMOX1 in rats with sepsis.

Epigenetics and the vaginal microbiome: influence of the microbiota on the histone deacetylase level in vaginal epithelial cells from pregnant women

Histone deacetylase (HDAC) influences the acetylation status of histones at gene promotor loci, providing an epigenetic mechanism that regulates gene expression. We determined if variations in the composition of the vaginal microbiome in pregnant women were associated with alterations in the level of HDAC1 in vaginal epithelial cells and whether this influenced the concentration of compounds present in vaginal fluid. Vaginal epithelial cells were obtained from 150 women in their first trimester of pregnancy, lysed and assayed for HDAC1 by ELISA. Composition of the vaginal microbiome was determined by classification of sequences amplified from the V1-V3 region of bacterial ribosomal 16S rRNA genes. Vaginal secretions were assayed for total protein, matrix metalloproteinase (MMP)-8, the 70kDa heat shock protein (hsp70) and the D- and L-lactic acid isomers. Lactobacilli were numerically dominant in 119 (79.3%) of the women, with Lactobacillus crispatus being the most prevalent (45.3% of women). Gardnerella was the most prevalent non-Lactobacillus species (10.7% of women). The median HDAC1 level in epithelial cells was 6.1 ng/mL when lactobacilli predominated vs. 20.5 ng/mL when non-lactobacilli were dominant (P=0.0039). Levels were lowest when L. crispatus was dominant (3.8 ng/mL) and highest with Streptococcus dominance (38.1 ng/mL). The concentration of HDAC1 was negatively correlated with the D-lactic acid level (P=0.0183) and positively correlated with concentrations of MMP-8 and hsp70 (P<0.0001) in the vaginal fluid. We propose that the composition of the vaginal microbiome and level of D-lactic acid, by influencing the HDAC1 level in vaginal epithelial cells, may epigenetically contribute to variations in the concentration of compounds in vaginal fluid.

PO-105 Exercise regulates HMGB1 / TLR4 / NF- κ B pathway by H2S to improve OJ intestinal injury

Objective To study the effect of aerobic exercise on the damage of intestinal mucosal barrier function caused by obstructive jaundice（OJ）and to explore its mechanism of action.&#x0D; Methods 50 male KM mice were randomly divided into 5 groups: sham operation group (S), model group (M), exercise group (TM), DL-Propargylglycine + exercise (PT) group and sodium hydrosulfide + exercise (NT) group.In addition to the S group which are in the common bile duct to the abdominal wall hanging 48 hours to build mouse obstructive jaundice model. In the PT group, PAG (40 mg/kg) was intraperitoneally injected 7 days after surgery; NaHS (50 μmol/kg) was intraperitoneally injected in the NT group 7 days after surgery; TM group, NT group and PT group were graded at 0%, and the speed was 10m/min no weight training (30min/day).After 6 weeks, HE staining was used to observe the morphological changes of the intestinal mucosa.Biochemical analysis was used to detect the concentration of hydrogen sulfide (H2S) in blood and ileum, and total bilirubin (TBIL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) Liver function, diamine oxidase (DAO), D-lactic acid intestinal barrier function biochemical index; qRT-PCR and immunohistochemical staining were used to observe the expression changes of H2S-mediated related channel mRNA and protein（HMGB1, TLR4 and NF-Kbp6）in intestinal tissues.&#x0D; Results HE staining showed that the intestinal mucosa of group M was atrophied and the villus was broken.Compared with M group, the intestinal mucosa arrangement in TM group was relatively regular. Compared with TM group, intestinal mucosa atrophy in PT group, fluff hair loss, sparseness and disorder, partial mucosa The layer was separated from the lamina propria and the gland was severely damaged. The intestinal mucosa of the NT group was relatively regular, and the changes of intestinal mucosa atrophy were restored. Serum test results showed that H2S levels were higher in the TM group than in the M group; compared with the TM group, the PT group decreased and the NT group increased. DAO level: The TM group was lower than the M group; compared with the TM group, the PT group was elevated and the NT group was decreased. Changes in serum D-lactic acid levels were similar to DAO. The results of qRT-PCR and immunohistochemical staining showed that the expressions of HMGB1, TLR4 and NF-Kbp6 mRNA and protein in the intestinal tissues of mice in TM group were significantly lower than those in M group and PT group, and the mRNA and protein expression levels in NT group were the lowest.&#x0D; Conclusions Aerobic exercise inhibits the HMGB1 / TLR4 / NF-κB signaling pathway through the H2S / CSE system, thereby exerting a protective effect on the intestinal mucosal barrier.

Effect of Simvastatin on the Intestinal Rho/ROCK Signaling Pathway in Rats With Sepsis

BackgroundSimvastatin may alleviate the intestinal barrier dysfunction induced by sepsis. This study aimed to investigate the role of the Ras homolog (Rho)/Rho-associated coiled-coil forming protein kinase (ROCK) signaling pathway in the intestinal barrier of simvastatin-treated rats with sepsis. Materials and MethodsMale Wistar rats were pretreated with simvastatin (0.2 μg/g of body weight) for 1 week before cecal ligation and puncture. Twenty-four hours after cecal ligation and puncture, the condition of bacterial translocation was evaluated. Plasma levels of intestinal fatty acid binding protein, D-lactic acid and inflammatory factors, and oxidative stress in the intestine were determined. The intestinal injury scores, as well as the protein levels of Rho, ROCK1, and tight junction proteins ZO-1 and occludin were analyzed. ResultsTreatment with simvastatin alleviated the sepsis-induced increases in the plasma concentration of intestinal fatty acid binding protein and D-lactic acid, as well as the number of colony-forming units in the bacterial culture of the blood, liver, spleen, and kidney. In addition, simvastatin effectively reduced the intestinal levels of tumor necrosis factor α, interleukin-6, high-mobility group box 1, and malondialdehyde and increased the activity of superoxide dismutase in rats with sepsis. Staining with hematoxylin and eosin showed that severe intestinal injury occurred in the sepsis group, which was reduced by the treatment of simvastatin. Furthermore, the expression of Rho and ROCK1 was significantly downregulated and the protein expression levels of ZO-1 and occludin were significantly increased in simvastatin-treated rats (P < 0.05). ConclusionsSimvastatin can ameliorate the intestinal barrier dysfunction caused by sepsis by inhibiting the Rho/ROCK signaling pathway and reducing the levels of inflammatory factors and oxidative stress in the intestine, which also increase the expression of tight junction proteins.

Lactobacillus frumenti Facilitates Intestinal Epithelial Barrier Function Maintenance in Early-Weaned Piglets.

Increased intestinal epithelial barrier function damages caused by early weaning stress have adverse effects on swine health and feed utilization efficiency. Probiotics have emerged as the promising antibiotic alternatives used for intestinal barrier function damage prevention. Our previous data showed that Lactobacillus frumenti was identified as a predominant Lactobacillus in the intestinal microbiota of weaned piglets. However, whether the intestinal epithelial barrier function in piglets was regulated by L. frumenti is still unclear. Here, piglets received a PBS vehicle or PBS suspension (2 ml, 108 CFU/ml) containing the L. frumenti by oral gavage once a day during the period of 6–20 days of age prior to early weaning. Our data demonstrated that oral administration of L. frumenti significantly improved the intestinal mucosal integrity and decreased the serum endotoxin and D-lactic acid levels in early-weaned piglets (26 days of age). The intestinal tight junction proteins (including ZO-1, Occludin, and Claudin-1) were significantly up-regulated by L. frumenti administration. The serum immunoglobulin G (IgG) levels, intestinal secretory immunoglobulin A (sIgA) levels, and interferon-γ (IFN-γ) levels were significantly increased by L. frumenti administration. Furthermore, our data revealed that oral administration of L. frumenti significantly increased the relative abundances of health-promoting microbes (including L. frumenti, Lactobacillus gasseri LA39, Parabacteroides distasonis, and Kazachstania telluris) and decreased the relative abundances of opportunistic pathogens (including Desulfovibrio desulfuricans and Candida humilis). Functional alteration of the intestinal bacterial community by L. frumenti administration was characterized by the significantly increased fatty acids and protein metabolism and decreased diseases-associated metabolic pathways. These findings suggest that L. frumenti facilitates intestinal epithelial barrier function maintenance in early-weaned piglets and may be a promising antibiotic alternative used for intestinal epithelial barrier function damage prevention in mammals.

Safety of lactic acid and calcium lactate when used as technological additives for all animal species.

The additives under assessment are lactic acid and calcium lactate. In 2015, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) delivered an Opinion on the safety and efficacy of lactic acid and calcium lactate. The FEEDAP Panel could not conclude on the safety of lactic acid in pre‐ruminants and poultry. Following this opinion, the European Commission gave the possibility to the applicant to submit complementary information in order to complete the assessment on the safety for all animal species. Based on the studies submitted in chickens for fattening and laying hens, no safe concentration of lactic acid and calcium lactate in complete feed for these species could be identified. Owing to the absence of data on tolerated dietary levels of d‐lactic acid, no conclusion on the safety of lactic acid in milk replacer for pre‐ruminants is possible. Since a safe concentration of lactic acid (and calcium lactate) was established only for pigs and cattle, and not for a third major animal species, no extrapolation to any other species is possible.

A study of the damage of the intestinal mucosa barrier structure and function of Ctenopharyngodon idella with Aeromonas hydrophila.

The aim of this study is to explore the effect of Aeromonas hydrophila on the intestinal mucosal barrier structure and intestinal permeability in grass carp (Ctenopharyngodon idella). Histopathological examinations showed that A. hydrophila induced severe intestinal lesions, including inflammatory cell infiltration and intestinal villus fusion and swelling. Messenger RNA (mRNA) expression of the inflammatory cytokines TNF-α, IL-1β, IL-8, IL-10 and MyD88 was significantly increased after infection with A. hydrophila. The permeability of intestinal mucosa was determined using Evans blue (EB) and D-lactic acid. The results indicated that the levels of EB and serum D-lactic acid were significantly increased after infection with A. hydrophila (p<0.05). Our results also indicated that the intestinal mucosal barrier injury induced by A. hydrophila infection was closely associated with the expression of the tight junction (TJ) protein zonula occludens-1 (ZO-1), occludin, claudin b and claudin c as well as the activity of Na+, K+-ATPase and Ca2+, Mg2+-ATPase. Lower mRNA levels of occludin and lower Na+, K+-ATPase and Ca2+, Mg2+-ATPase activity in the intestines were observed after challenge. ZO-1 and claudin c were significantly increased 24h after infection with A. hydrophila. The most interesting finding was that claudin b also significantly increased 24h after challenge and then decreased to lower levels at 72, 120 and 168h post-infection compared to the PBS-treated control group. The results demonstrated that grass carp infection with A. hydrophila induced intestinal inflammation and impaired the structure and function of the intestinal mucosal barrier.

Jinhong Tablet Reduces Damage of Intestinal Mucosal Barrier in Rats with Acute Biliary Infection via Bcl-2/Bax mRNA and Protein Regulation.

Objective To explore the effects and mechanism of Jinhong Tablet on intestinal mucosal barrier function and SIRS in rats with acute biliary infection. Methods 36 SD male rats were divided into three groups: sham operation (control), acute biliary infection (ABI) model, and Jinhong Tablet (Jinhong) group. Jinhong group were force-fed with Jinhong Tablet, while the other two groups received oral saline. At days 3 and 5, morphological changes of intestinal mucosa were assessed. Serum diamine oxidase (DAO), D-lactate, and endotoxin levels were measured. And the genes bcl-2 and bax in intestinal tissues were tested by real-time PCR and Western blotting. Results Intestinal damage was significantly less severe in Jinhong group compared with ABI group, as indicated by Chiu's scoring, TUNEL analysis, and serum DAO, D-lactic acid, and endotoxin levels. Additionally, the expression of bax mRNA and protein was decreased and the ratio of bcl-2/bax mRNA and protein was increased compared with ABI group. Conclusion Jinhong Tablet had a positive intervention on acute biliary infection through improving inflammation and intestinal mucosal barrier, inhibiting excessive apoptosis of intestinal epithelial cells via bax and bcl-2 gene, and protein regulation.

385: D-lactic Acid and Matrix Metalloproteinase-2 (MMP-2) levels in vaginal secretions predict cervical length in mid-trimester pregnant women

385: D-lactic Acid and Matrix Metalloproteinase-2 (MMP-2) levels in vaginal secretions predict cervical length in mid-trimester pregnant women

Intestinal barrier dysfunction and its related factors in patients with sepsis

Objective: To investigate the relationship between related factors of intestinal barrier dysfunction in patients with sepsis or septic shock and severity of the condition. Methods: A prospective observational study was conducted in 31 sepsis patients, 28 septic shock patients, and 21 postoperative patients without sepsis (control group) who were admitted to intensive care unit (ICU) of General Hospital of Ningxia Medical University between November 2015 and June 2016. Blood samples were collected from the patients within 24 hours following admission to ICU. D-lactic acid and endotoxin levels were measured by enzymatic method, serum high-sensitivity C-reactive protein (hsCRP) level by immune scatter turbidimetry. An arterial blood gas (ABG) measurement was carried out every 8 hours within the first 24 hours after admission to ICU, and average arterial blood lactate levels were calculated. Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score, Sequential Organ Failure Assessment (SOFA) score of the patients within 24 hours following ICU admission were recorded. The patients with sepsis or septic shock were followed up for 28 days after admission to ICU, and divided into survival group (n=44) and death group (n=15). The patients with sepsis or septic shock were divided into two groups according to the site of infection, i. e. intra-abdominal infection group (n=37) and extra-abdominal infection group (n=22). Results: (1) In the control, sepsis, and septic shock groups, D-lactic acid [mg/L, M(P25, P75) ] were 11.68(7.49, 14.92), 19.78 (12.25, 34.85), and 32.45 (16.03, 46.95), respectively; endotoxin levels [U/L, M(P25, P75)] were 10.60(7.59, 13.39), 16.12(10.09, 20.23), and 17.31(14.09, 23.77), respectively. The levels of serum D-lactic acid and endotoxin in the patients with sepsis or septic shock were significantly higher than those in the control group (all P<0.01); while no statistically significant differences existed in these two indices between the sepsis and septic shock groups (both P>0.05). There were no statistically significant differences in serum D-lactic acid and endotoxin levels between the intra-abdominal infection group and the extra-abdominal infection group [20.07(14.70, 38.97)vs 21.65 (14.53, 56.56)mg/L; 17.23(13.38, 20.85)vs 17.17(9.93, 20.81)U/L; both P>0.05]. There were no statistically significant differences in levels of serum D-lactic acid and endotoxin between the survival group and the death group [21.65(15.11, 39.00) vs 19.78(14.41, 80.93)mg/L; 17.09(12.62, 20.42) vs 19.26(13.22, 26.27)U/L, both P>0.05]. (2) In the sepsis and septic shock patients, serum D-lactate level was significantly related to mean arterial blood lactate concentration, APACHE Ⅱ score, and SOFA score in the first 24 hours after admission to ICU (r=0.499, 0.447, 0.469, all P<0.01); serum endotoxin level was correlated with hsCRP, APACHE Ⅱ score, and SOFA score (r=0.224, 0.388, 0.393, all P<0.05). (3) Multivariate linear regression analysis showed that D-lactic acid level was independently associated with average arterial blood lactate concentration as well as with SOFA score(R2=0.34, F=19.91, P<0.01), and endotoxin was independently associated with only SOFA score(R2=0.14, F=12.68, P<0.01). Conclusions: Regardless of the site of infection, patients with sepsis or septic shock often have intestinal barrier injury, which is correlated with the severity of disease, but does not independently affect patient outcome. Tissue hypoperfusion in the early stage of sepsis may be one of the causes of intestinal barrier injury.

Effects of glutamine on markers of intestinal inflammatory response and mucosal permeability in abdominal surgery patients: A meta-analysis.

The present meta-analysis was carried out to determine whether supplementation with glutamine (Gln) would reduce the intestinal inflammatory response and mucosal permeability in patients undergoing abdominal surgery. The PubMed, EMBASE, Web of Science, and The Cochrane Library databases were searched for randomized controlled trials on the effects of supplementation with Gln, and published from August, 1966 to June 2014. Inclusion criteria for the meta-analysis were: i) Study design was a randomized controlled trial, ii) study included patients undergoing abdominal surgery, iii) study patients received a supplementation with Gln peptide (Ala-Gln or Gly-Gln) whereas control patients did not use any supplements, and iv) study outcomes included inflammatory markers [C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), and interleukin (IL)-6, and IL-2 receptor] and markers of intestinal permeability [lactulose/mannitol, diamine oxidase, D(−)lactic acid, and endotoxin]. Qualities of controlled trials were assessed using the Jadad score. Meta-analyses were performed with fixed- or random-effect models depending on the heterogeneity of studies. There were 21 trials meeting the inclusion criteria. The meta-analysis revealed that the levels of CRP, TNF-α, and IL-6 in patients supplemented with Gln were significantly lower than those in control patients, whereas the levels of IL-2 receptor were increased by Gln supplementation. Gln also significantly decreased the lactulose/mannitol ratio, the levels of diamine oxidase and endotoxin, and tended to decrease the levels of cyclic D-lactic acid. In conclusion, Gln appears to effectively reduce the inflammatory response and intestinal mucosal permeability in patients after abdominal surgery.

Protective effects of heparin on early septic shock of intestinal mucosal injury in dogs

Objective Determination of plasma markers of D-lactic acid and I-FABP and analyzed to verify the early septic shock in dogs and intestinal fatty acid binding protein expression levels of D-lactate changes and improvements to septic shock heparin early jejunal villus microcirculation in dogs and intestinal fatty acid binding protein and the expression of D-lactic acid levels, confirmed the protective effect of heparin on intestinal mucosal injury. Methods Beagles early septic shock by intravenous injection of LPS Methods. The 40 animals were randomly random number divided into control group, LPS group, the basic treatment group, heparin group. Stable after 1 hour, i. e. the point T0, to modeling LPS 1 hour, i. e. at T1, after 5 hours was observed sequential T2, T3, T4, T5, T6 point. Observed at the corresponding time points (T1, T3, T6 time points) canine blood collected by 1 500 rpm, 15 minutes after centrifugation, the plasma to the EP tube into aliquots stored at -80 ℃, measured by ELISA intestinal fatty acid binding protein (IFABP) and D-lactate (D-lac). Results heparin group IFABP (21.6±2.3) pg / mL LPS comparison group IFABP (100.48±8.42) pg / mL will be significantly decreased, with statistical significance, P 0.05, on the basis of adequate volume resuscitation on heparin therapy, I-FABP expression levels can be significantly reduced. Conclusions The presence of septic shock early intestinal mucosal microcirculation and intestinal mucosa. Under the premise of its full capacity of recovery, heparin early septic shock in dogs with intestinal mucosal microcirculation improvement, thereby protecting the intestinal mucosal barrier function. Key words: Sepsis / Septic shock; Heparin; Intestinal mucosa; Microcirculation; Lipopolysaccharides; Intestinal binding protein; D-lactic acid; Orthogonal polarization spectral imager

Glutamine decreases intestinal mucosal injury in a rat model of intestinal ischemia-reperfusion by downregulating HMGB1 and inflammatory cytokine expression.

Intestinal ischemia-reperfusion (IR) is a common clinical pathophysiological process that is common in severe trauma, major surgery, and in post-resuscitation. Glutamine (Gln) reduces intestinal IR injury, however, its mechanism of action remains to be determined. High mobility group box 1 (HMGB1) protein, nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), and interleukin-1 (IL-1) are mediators involved in the pathophysiology of intestinal IR injury. The aim of the present study was to investigate the effects of Gln on the intestinal mucosa of HMGB1 expression following IR to determine whether Gln relieved intestinal IR injury in the intestinal mucosal barrier. Forty-eight Sprague-Dawley rats were included in the present study. A model of intestinal ischemia-reperfusion injury was established by clamping the superior mesenteric artery of the rats to cause ischemia, followed by restoring blood flow. The animals were randomly divided into the control (n=24) and the Gln (n=24) groups for the experiments. The two groups of rats were given enteral nutrition with equal heat, nitrogen (heat 125.4 kJ/kg/day, nitrogen 0.2 g/kg/day). The Gln group of rats was fed with enteral nutrition plus 3% Gln, while the control rats were fed with enteral nutrition plus 3% soybean protein. After 7 days, the HMGB1 and plasma levels of NF-κB, TNF-α, IL-1, Gln, D-lactic acid and diamine oxidase (DAO) were observed. The changes in the morphology of intestinal mucosa were observed using electron microscopy. The plasma levels of TNF-α, IL-1, D-lactic acid and DAO, and the level of HMGB1, NF-κB, TNF-α and IL-1 in intestinal mucosa were significantly higher after IR (p<0.05), while the plasma level of Gln was lower in the two groups. In the control group, the plasma level of IL-1, TNF-α, DAO and D-lactic acid, and that of HMGB1, NF-κB, TNF-α, and IL-1 in intestinal mucosa were significantly higher, while the plasma level of Gln was lower than that prior to modeling on the 3rd and 7th days of the experiment. In the Gln group, the plasma level of IL-1, TNF-α, DAO and D-lactic acid, and that of HMGB1, NF-κB, IL-1, and TNF-α in intestinal mucosa were significantly higher (p<0.05) compared to the control on the 3rd and 7th days of the experiment. By contrast, after the 7th day, the plasma level of IL-1, TNF-α, DAO and D-lactic acid, and the level of HMGB1, NF-κB, IL-1, TNF-α in intestinal mucosa were significantly lower in the Gln group, while the plasma level of Gln was significantly higher than those in control group and after IR on the 7th day of the experiment. Additionally, the structure of villi and recess was damaged, villi was sparse and short, and considerable inflammatory cell influx embellished the lamina propria, lymphangiectasia, and edema after IR. On the 7th day, compared to after IR, the intestinal villi and recess structure of the controls was significantly restored in the Gln group. In conclusion, Gln repaired the intestinal mucosal injury in IR by reducing the expression of HMGB1 and inflammatory cytokines, and reducing the permeability of the intestinal mucosa.

Intestinal Conditioning After Cardiac Arrest: The Use of Normothermic Extracorporeal Membrane Oxygenation in the Non-Heart-Beating Animal Model.

The effect of normothermic extracorporeal membrane oxygenation (NECMO) on small bowel preservation in a clinically relevant large animal model of expected donation after cardiac death (eDCD) was evaluated. Thirty domestic crossbred donor pigs were divided into five groups. The first group served as the live donation (LD) group, the second group served as the donation after cardiac death (DCD) group, and the remaining were further assigned into three subgroups: E1 group (1 h NECMO support), E3 group (3 h NECMO support), and E5 group (5 h NECMO support). Pathology, electron microscopy, energy metabolism, cell apoptosis, and tight junction (TJ) protein expression level of intestinal mucosa and the level of plasma d-lactic acid were evaluated in normal, cardiac death and at the end of extracorporeal support, respectively. The mean arterial pressure and PaO2 were maintained over 60 and 267 mmHg during NECMO support, respectively. One hour of extracorporeal support could improve the energy status in intestines of the DCD group. Although the histologic damage and apoptosis of the E1 group had no significant difference with those of the LD and DCD groups (P > 0.05), the levels of intestinal mucosa TJ protein decreased (P < 0.05), and plasma d-lactic acid increased progressively (P < 0.05). With the extension of extracorporeal support, the degree of intestinal mucosa damage and intestinal permeability gradually increased, as well as the content of adenosine triphosphate in intestinal mucosa. The normothermic extracorporeal support for 1 h in DCD is beneficial for improving the energy status and viability of the bowel. However, the integrity of intestinal mucosa was destroyed gradually as extracorporeal support time went by. And the activation of intestinal epithelial apoptosis and hyperoxia might be the factors that lead to intestinal mucosa injury.

Impaired function of the intestinal barrier in a novel sub-health rat model.

Sub-health is a state featuring a deterioration in physiological function between health and illness, and the sub-health condition has surfaced as life-threatening in humans. The aim of the present study was to establish a sub-health model in rats, and investigate the function of the intestinal barrier in the sub-health rats and rats following intervention. To establish a sub-health model, the rats were subjected to a high-fat and sugar diet, motion restriction and chronic stress. Their serum glucose and triglyceride levels, immune function and adaptability were then measured. The levels of diamine oxidase and D-lactic acid in the plasma were analyzed as markers of the intestinal permeability. The protein and mRNA expression levels of anti-apoptotic YWHAZ in the colonic tissue was detected using immunohistochemical and reverse transcription-quantitative polymerase chain reaction analyses In the present study, the sub-health rat model was successfully established, and sub-health factors increased the intestinal permeability and reduced the expression of YWHAZ. Providing sub-health rats with normal living conditions did not improve the function of the intestinal barrier. In conclusion, the results of the present study demonstrated that intestinal disorders in the sub-health rat model may result from the damage caused by reduce intestinal barrier function as well as the decreased expression levels of YWHAZ. Additionally, rats in the sub-health condition did not recover following subsequent exposure to normal living conditions, suggesting that certain exercises or medical intervention may be necessary to improve sub-health symptoms.

Effects of lactadherin on plasma D-lactic acid and small intestinal MUC2 and claudin-1 expression levels in rats with rotavirus-induced diarrhea.

The aim of the present study was to investigate the effects of lactadherin on plasma D-lactic acid and small intestinal mucin (MUC) 2 and claudin-1 expression levels in rats with diarrhea induced by rotavirus (RV) infection. A total of 75 seven-day-old healthy Sprague-Dawley rats were randomly divided into the following five groups: Control (C), RV infection (RVI), lactadherin before rotavirus infection (LBRI), lactadherin after rotavirus infection (LARI), and blank (B). On day 4 of artificial feeding, the rats in groups RVI, LBRI and LARI were intragastric administered 1×106 PFU RV; whereas the rats in groups C and B were intragastrically administered an equal volume of maintenance solution from the RV supernatant and normal saline, respectively. In the LBRI and LARI groups, rats received daily intragastric administration of 0.25 mg lactadherin for three days prior to and following infection with RV, respectively. The course of diarrheal symptoms was observed in each group and samples were collected on days 1, 4, and 7 post-infection in order to determine the mucosal morphology, plasma D-lactic acid levels and the expression levels of MUC2 and the intracellular junction protein, claudin-1, in the small intestine. On day 4 post-infection, the rats in group RVI demonstrated severely damaged small intestines and typical diarrheal characteristics, as detected by light microscopy; whereas rats in groups LBRI and LARI demonstrated intact small intestinal villi with partial vacuolation of epithelial cells and changes in the position of their nuclei. Electron microscopy demonstrated that the rats in the RVI group had sparse, shortened, disordered intestinal microvilli and widened intercellular junctions; whereas those in groups LBRI and LARI had long intestinal microvilli sparser compared with groups B and C and slightly widened intercellular junctions. Plasma D-lactic acid levels were increased in groups RVI, LBRI and LARI, as compared with groups B and C, and the greatest levels were detected in the RVI group on days 1, 4 and 7 post-infection. In addition to maintaining intestinal permeability, lactadherin enhanced the expression levels of MUC2 and reduced the expression of claudin-1; therefore, further protecting the intestinal epithelial barrier, which may contribute to the prevention and treatment of diarrhea induced by infection with RV.

Effects of early enteral nutrition on the gastrointestinal motility and intestinal mucosal barrier of patients with burn-induced invasive fungal infection.

Objective:To evaluate the effects of early enteral nutrition on the gastrointestinal motility and intestinal mucosal barrier of patients with burn-induced invasive fungal infection.Methods:A total of 120 patients with burn-induced invasive fungal infection were randomly divided into an early enteral nutrition (EN) group and a parenteral nutrition (PN) group (n=60). The patients were given nutritional support intervention for 14 days, and the expression levels of serum transferrin, albumin, total protein, endotoxin, D-lactic acid and inflammatory cytokines were detected on the 1st, 7th and 14th days respectively.Results:As the treatment progressed, the levels of serum transferrin, albumin and total protein of the EN group were significantly higher than those of the PN group (P<0.05), while the levels of serum endotoxin and D-lactic acid of the form group were significantly lower (P<0.05). After treatment, the expression levels of IL-6 and TNF-α were decreased in the EN group, which were significantly different from those of the PN group (P<0.05). During treatment, the incidence rates of complications such as abdominal distension, diarrhea, sepsis, nausea, vomiting and gastric retention were similar. The mean healing time of wound surface was 9.34±0.78 days in the EN group and 12.46±2.19 days in the PN group, i.e. such time of the former was significantly shorter than that of the latter (P<0.05).Conclusion:Treating patients having burn-induced invasive fungal infection by early enteral nutrition support with arginine can safely alleviate malnutrition and stress reaction, strengthen cellular immune function and promote wound healing, thereby facilitating the recovery of gastrointestinal motility and the function of intestinal mucosal barrier.

Shenfu injection alleviates intestine epithelial damage in septic rats

BackgroundShenfu injection (SFI) promotes tissue microcirculation and oxygen metabolism. We aimed to assess its effects on intestinal epithelial damage in septic rats. MethodsFifty Sprague-Dawley rats were randomly divided into sham operation (Sham), sepsis (cecal ligation and puncture [CLP]), and SFI (low-dose, middle-dose, high-dose) groups (n = 10). For Sham animals, the abdominal cavity was opened and closed. For other groups, severe sepsis was induced by CLP. After surgery, saline (Sham and CLP rats) and SFI (treatment groups) were administered intraperitoneally. Samples were collected 12 hours after injection. Serum tumor necrosis factor α, diamineoxidase, and d-lactate levels and ileal mucosal damage and ultrastructural change, as well as protein and messenger RNA expression of tight junction markers, including Claudin-3 and zonula occludens protein-1 in ileal mucosa's epithelial cells, were assessed. All animal experiments were carried out under aseptic conditions. ResultsCompared with Sham animals, serum tumor necrosis factor α, DAO, and d-lactic acid levels in CLP animals were significantly higher; the ileal mucosal damage was more severe; and the expression levels of tight junction markers were significantly decreased. These indexes were significantly improved in SFI groups, in a concentration-dependent manner, compared with CLP rats. Sham animals displayed orderly arranged ileal mucosal villi, continuous tight junctions between epithelial cells, intact organelles, and microvilli. Compared with CLP animals (with obvious damage in these structures), an overt improvement was observed in SFI groups, especially in the high-dose SFI group, with tight junctions clearly visible between epithelial cells. ConclusionsShenfu injection significantly alleviates intestinal epithelial damage in septic rats, in a dose-dependent manner.