Ameliorative effect of pedunculoside on sepsis-induced acute lung injury, inflammation and pulmonary fibrosis in mice model via suppressing AKT/NF-κB pathway.

BACKGROUND/OBJECTIVES: Sepsis-induced acute lung injury (ALI) is the typical complications of sepsis with a high global incidence and mortality. Inhibition of inflammatory response is a crucial and effective strategy for sepsis-induced ALI. Pedunculoside (PE) has been shown to have an anti-inflammatory effect on various diseases. However, the effect and mechanism of PE on sepsis-induced ALI remain unknown. MATERIALS/METHODS: A mice model of sepsis-induced ALI was constructed by cecal ligation and puncture (CLP). The effect of PE on the CLP-induced mice were assessed using pathological staining, terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL), reverse transcription quantitative polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay (ELISA) and western blot assays. RESULTS: PE reduced pathological symptoms and scores, apoptosis and the W/D ratio of lung tissues in CLP-induced mice. Besides, PE decreased the level of interleukin (IL)-1ß, IL-6 and tumor necrosis factor (TNF)-α), pulmonary fibrosis and the expression of fibrosis markers. Mechanically, PE inhibited AKT/NF-κB signaling in CLP-induced mice. Activation of AKT/NF-κB pathway abolished the ameliorative effect of PE on the pathological symptoms, the release of inflammatory factors and pulmonary fibrosis of CLP-induced mice. CONCLUSION: PE improved inflammation and pulmonary fibrosis by inhibiting AKT/NF-κB pathway in CLP-induced mice.

Predictive Model of Cerebral Vasospasm in Subarachnoid Hemorrhage Based on Regression Equation.

In order to explore the regression equation for the prediction model of subarachnoid hemorrhage and cerebral vasospasm, the nomogram prediction model of SCVS occurrence was established. This study is a retrospective analysis of 125 cases of aSAH admitted to a hospital; the patients were divided into SCVS group and non-SCVS group. Select SIRI as a simple and reliable marker of inflammation, analyze its correlation with SCVS and its predictive value, and analyze the predictive value of SIRI to SCVS through ROC curve. Based on the SIRI inflammation level and other related risk factors, a nomogram prediction model for the occurrence of SCVS was built. The experimental results show that the SIRI level of patients in the SCVS group was significantly higher than that of the non-SCVS group, and logistic regression analysis found that SIRI is an independent risk factor for SCVS. SIRI = 3.63 × 109/L is the best cutoff value for diagnosing the occurrence of SCVS. When TC = 2.24 mmol/L and SIRI = 3.63 × 10%/L, its Youden Index is the largest (0.312, 0.296) and is the best cutoff value for predicting the occurrence of SCVS; at the same time, its prediction accuracy (area under the ROC curve (AUC)), sensitivity, specificity, the positive predictive value, and negative predictive value are 0.743, 72.70%, 80.10%, 77.53%, and 94.24% and 0.725, 70.60%, 76.90%, 73.49%, and 93.59%. Nomogram prediction model establishment and evaluation combined with the results of multifactor analysis are used to build an individual nomogram prediction model. The model has good prediction consistency (C-index = 0.685, P < 0.01). ROC analysis results showed that the model that combined SIRI and other standard variables (AUC = 0.896, 95% CI was 0.803-0.929, P < 0.001) was better than the model that did not combine SIRI (AUC = 0.859, 95% CI was 0.759-0.912, P < 0.001) and the model based only on SIRI (AUC = 0.725, 95% CI was 0.586-0.793, P = 0.001) has better predictive value for SCVS. Joint SIRI will optimize the prediction performance of the nomogram model and improve the early recognition and screening capabilities of SCVS.

Identification of an Immune-Related Biomarker Model Based on the CircRNA-Associated Regulatory Network for Esophageal Carcinoma.

Esophageal carcinoma (ESCA) is one of the most frequent types of malignant tumor that has a dismal prognosis. This research applied datasets aimed from the GEO and TCGA to create a prognostic signature for forecasting the clinical outcome of ESCA patients on the basis of a circRNA-associated regulatory network. Methods. A regulatory network associated with ESCA was established based on transcriptome data of circRNAs, miRNAs, and mRNAs. Functional annotations were implemented to further explore the mechanism of ESCA. Cox relative regression method was applied to create a risk signature. Besides, the immune microenvironment of the signature was investigated by utilizing the CIBERSORT algorithm. Results. Based on 27 DEcircRNAs, 65 DEmiRNAs, and 780 DEmRNAs, the circRNA-miRNA-mRNA network was finally set up. Functional enrichment unearthed that the regulatory network might participate in phosphorylation negative regulation, MAPK pathway, and PI3K/AKT pathway. This study established a risk scoring signature based on the seven immune-related genes (IRGs) (MARP14, RASGR1, PTK2, HMGB1, DKK1, RARB, and IGF1R), which was validated for its reliability. A stable and accurate nomogram combining immune-related risk scores with clinical features was constructed. Furthermore, we observed that the risk model was also related to the immunocyte infiltration. Conclusion. Our study successfully created a circRNA-associated regulatory network and further developed an immune-related model to forecast the clinical outcome of ESCA patients as well as to assess their immune status.

The prognostic value of routine coagulation tests for patients with heat stroke.

OBJECTIVE: To evaluate the prognostic value of routine coagulation tests for patients with heat stroke. METHODS: This was a multi-center retrospective study. Patients who arrived at the hospital <24 h after the onset of Heat Stroke (HS) were included. The routine coagulation variables were detected within 24 h after the onset, including the lowest platelet count (PLC). RESULTS: 60-day mortality rate was 20.9%. The median Prothrombin Time-International Normalized Ratio (PT-INR) of the non-surviving patients was significantly higher than that of the survivors (P < 0.01). The median Activated Partial Thromboplastin Time (APTT) in non-surviving patients was significantly higher than in the surviving patients (P < 0.01). A Cox regression analysis revealed that 60-day mortality was associated with PT-INR (P = 0.032) and APTT (P = 0.004). The optimal PT-INR point for predicting 60-day mortality rate was 1.7. The optimal APTT point for predicting 60-day mortality was 51.45. Patients with increased PT-INR (≥1.7) levels had, overall, a significantly reduced survival time (P < 0.01). Patients with elevated APTT (≥51.45) also had a decrease in survival time (P < 0.01). The prognostic scoring, with increased PT-INR (≥1.7) and prolonged APTT (≥51.45) at one point each, was also demonstrated to be useful in predicting 60-day mortality. Patients whose temperature fell to 38.9 °C within 30 min had significantly lower levels of PT-INR and APTT within 24 h than those who took longer to cool down. CONCLUSIONS: A prolonged APTT and elevated PT-INR within 24 h are independent prognostic factors of 60-day mortality in HS.

Omega-3 polyunsaturated fatty acids inhibit the increase in cytokines and chemotactic factors induced in vitro by lymph fluid from an intestinal ischemia-reperfusion injury model.

OBJECTIVE: To investigate the active factors and the intervention effect of ω-3 polyunsaturated fatty acids (PUFAs) during intestinal ischemia-reperfusion (I/R) injury, which causes the inflammation of monocytes-macrophages cultured in lymph fluid and stimulated with ω-3 PUFAs. METHODS: Forty-eight Sprague-Dawley male rats were randomly divided into the following two groups: A. (N + D) group and B. (I/R + D) group. The rats in the (N + D) group were drained of lymph for 180 min; the rats in the (I/R + D) group were subjected to 60 min ischemia by clamping the superior mesenteric artery followed by 120 min reperfusion and 180 min of lymph draining. Lymph fluid from each group was further divided into 4 subgroups, respectively: lymph group (A1, B1); eicosopentaenoic acid (EPA)-treated group (A2, B2); EPA + docosahexaeonic acid (DHA)-treated group (A3, B3); and DHA-treated group (A4, B4), then cultured monocyte-macrophage cell line. RESULTS: The levels of tumor necrosis factor-α, interleukin (IL)-1 ß, IL-6, soluble cell adhesion molecule-1, chemotactic factors macrophage chemoattractant protein-1, macrophage inflammatory protein-2, and high mobility group box protein 1 in the B1 group were significantly higher than in the A1 group. Importantly, addition of EPA, EPA + DHA, and DHA to the culture media significantly reduced the levels of the above-mentioned factors. Cell stimulation with EPA, EPA + DHA, and DHA also significantly decreased the expression of Toll-like receptor 4, nuclear factor-κB p65, macrophage chemoattractant protein-1, and macrophage inflammatory protein-2 with the combined treatment of EPA and DHA showing the strongest effect. CONCLUSIONS: The factors induced in lymph during intestinal I/R injury can cause inflammation in vitro. These data provide in vitro evidence that ω-3 PUFAs provide a protective effect by reducing the inflammatory response caused by intestinal I/R lymph. Moreover, the synergism of EPA and DHA had the greatest effect, which is possibly mediated through Toll-like receptor 4 and nuclear factor-κB p65.

Impact of intestinal ischemia/reperfusion and lymph drainage on distant organs in rats.

AIM: To investigate the impact of intestinal ischemia/reperfusion (I/R) injury and lymph drainage on distant organs in rats. METHODS: Thirty-two Sprague-Dawley male rats, weighing 280-320 g, were randomly divided into blank, sham, I/R, and ischemia/reperfusion and drainage (I/R + D) groups (n = 8). All rats were subjected to 60 min ischemia by clamping the superior mesenteric artery, followed by 120 min reperfusion. The rats in the I/R + D group received intestinal lymph drainage for 180 min. In the sham group, the abdominal cavity was opened for 180 min, but the rats received no treatment. The blank group served as a normal and untreated control. A chromogenic limulus assay kit was used for quantitative detection of serum endotoxin. The serum concentrations of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1ß, soluble cell adhesion molecules (sICAM-1), and high mobility group protein box 1 (HMGB1) were determined with an enzyme-linked immunosorbent assay kit. Histological evaluations of the intestine, liver, kidney, and lung were performed by hematoxylin and eosin staining and immunohistochemistry. HMGB1 protein expression was assayed by western blot analysis. RESULTS: The serum levels of endotoxin and HMGB1 in the I/R and I/R + D groups were significantly higher than those in the sham group (endotoxin, I/R and I/R + D vs sham: 0.033 ± 0.004 EU/mL, 0.024 ± 0.003 EU/mL vs 0.017 ± 0.009 EU/mL, respectively, P < 0.05; HMGB1, I/R and I/R + D vs sham: 5.473 ± 0.963 EU/mL, 4.906 ± 0.552 EU/mL vs 0.476 ± 0.406 EU/mL, respectively, P < 0.05). In addition, endotoxin and HMGB1 were significantly lower in the I/R + D group compared to the I/R group (P < 0.05). The serum inflammatory factors IL-6, IL-1ß, and sICAM-1 in the I/R and I/R + D groups were significantly higher than those in the sham group (IL-6, I/R and I/R + D vs sham: 41.773 ± 9.753 pg/mL, 19.204 ± 4.136 pg/mL vs 11.566 ± 2.973 pg/mL, respectively, P < 0.05; IL-1ß, I/R and I/R + D vs sham: 144.646 ± 29.378 pg/mL, 65.829 ± 10.888 pg/mL vs 38.178 ± 7.157 pg/mL, respectively, P < 0.05; sICAM-1, I/R and I/R + D vs sham: 97.360 ± 12.714 ng/mL, 48.401 ± 6.547 ng/mL vs 33.073 ± 5.957 ng/mL, respectively; P < 0.05). The serum TNF-α in the I/R group were significantly higher than in the sham group (45.863 ± 11.553 pg/mL vs 18.863 ± 6.679 pg/mL, respectively, P < 0.05). These factors were significantly lower in the I/R + D group compared to the I/R group (P < 0.05). The HMGB1 immunohistochemical staining results showed no staining or apparent injury in the blank group, and slight staining at the top of the microvillus was detected in the sham group. In the I/R group, both the top of villi and the basement membrane were stained for HMGB1 in most areas, and injury in the I/R + D group was less than that in the I/R group. HMGB1 expression in the liver, kidney, and lung of rats in the I/R + D group was significantly lower than the rats in the I/R group (P < 0.05). CONCLUSION: Lymph drainage could block the "gut-lymph" pathway, improve intestinal barrier function, and attenuate distant organ injury incurred by intestinal I/R.

The effects of n-3 PUFA and intestinal lymph drainage on high-mobility group box 1 and Toll-like receptor 4 mRNA in rats with intestinal ischaemia-reperfusion injury.

The aim of the present study was to investigate the impacts of n-3 PUFA and lymph drainage (D) on intestinal ischaemia-reperfusion (I/R) injury in rats. A total of forty-eight Sprague-Dawley male rats were randomly divided into three groups (n 16): normal diet (N), enteral nutrition (EN) and EN plus n-3 PUFA. Each group was further divided into lymph drainage (I/R+D) and non-drainage (I/R) sub-groups (n 8). After 5 d with different nutrition regimens, the rats were subjected to 60 min ischaemia by clamping the superior mesenteric artery, followed by 120 min reperfusion. At the same time, the rats in the I/R+D sub-groups were treated with intestinal lymph drainage for 180 min. Organs were harvested and we detected the cytokine, endotoxin, and expression of Toll-like receptor (TLR) 4 mRNA and its endogenous ligand high-mobility group box 1 (HMGB1). We found that the serum levels of HMGB1, inflammatory cytokine and endotoxin in the three I/R+D sub-groups were significantly lower than those in the N (I/R) and EN (I/R) sub-groups (P < 0·05). The activation of NF-κB and the expression of HMGB1 and TLR4 mRNA significantly increased in the jejunum, ileum, liver and lung after intestinal I/R injury, but notably lower in the I/R+D groups than those in I/R (P < 0·05). The injury degree and HMGB1 expression were decreased in the n-3 PUFA group than in the N and EN groups. We preliminarily concluded that nutrition with n-3 PUFA and/or intestinal lymph drainage may reduce HMGB1 and inflammatory cytokine in serum and lymph and inhibit the expression and signal transmission of TLR4 mRNA, thereby alleviating intestinal I/R injury in rats.

[Effects of lymphatic drainage and omega-3 polyunsaturated fatty acids on intestinal ischemia-reperfusion injury in rats].

OBJECTIVE: To investigate the effects of lymphatic drainage and omega-3 polyunsaturated fatty acid (omega-3PUFA) on high mobility group box 1 (HMGB1), inflammatory cytokines and endotoxin in rats with intestinal ischemia-reperfusion (I/R) injury. METHODS: A total of 72 SD rats were randomly divided into drainage-alone group, I/R group, ischemia-reperfusion plus drainage (I/R + D) group (n = 8 each) and 3 groups with 16 rats undergoing gastrostomy in each group: normal diet (N) group, enteral nutrition (EN) group and enteral nutrition & omega-3PUFA (PUFA) group. And they were further divided into 2 subgroups (n = 8). The rats in I/R and I/R + D groups were subjected to a 60-min ischemia follow by 120-min reperfusion injury of superior mesenteric artery. When the rats suffered I/R injury, intestinal lymph was drained for 180 min in the I/R + D group. The rats in the drainage-alone group received 180-min lymph drainage without I/R injury. After 5 days with different nutrition regimes, the models were established similarly. The rats in the I/R + D sub-groups were treated with intestinal lymph drainage for 180 min. The serum and lymph samples were collected post-operatively. Endotoxin was detected by a Limulus kit. The inflammatory cytokines and high mobility group box 1 (HMGB1) were analyzed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Endotoxin, inflammatory cytokines and lymphatic HMGB1 of lymphatic in the I/R + D group were higher than those in the drainage-alone group [all P < 0.05, IL-6: (30 +/- 8) pg/ml vs (20 +/- 6) pg/ml, endotoxin: (0.029 +/- 0.011) U/ml vs (0.008 +/- 0.005) U/ml]. The serum levels of endotoxin and inflammatory cytokines in the I/R + D group were lower than those in the I/R group (P < 0.05). The lymphatic levels of TNF-alpha (tumor necrosis factor-alpha) and HMGB1 in the N and EN groups were higher than those in the PUFA group[ TNF-alpha: (46 +/- 17) pg/ml, (54 +/- 16) pg/ml vs (28 +/- 9) pg/ml, HMGB1: (4.8 +/- 1.6) ng/ml, (5.3 +/- 1.8) ng/ml, (3.0 +/- 1.0) ng/ml, all P < 0.05)]. The serum levels of endotoxin, inflammatory cytokines and HMGB1 in the PUFA (I/R) group were lower than those in the N (I/R) group (P < 0.05). The levels of TNF-alpha and HMGB1 were lower in the PUFA (I/R + D) group than those in the N (I/R + D) group (both P < 0.05). CONCLUSION: Lymphatic drainage may reduce the levels of endotoxin, inflammatory cytokines and HMGB1 so as to alleviate the intestinal I/R injury. The intervention of omega-3PUFA has some protective effect through relieving inflammation.

[The sheltering effects of ω-3 polyunsaturated fatty acids and lymphatic drainage on distant organs after intestinal ischemia reperfusion injury in rats].

OBJECTIVE: To investigate the sheltering effects of ω-3 polyunsaturated fatty acid (ω-3PUFA) and lymphatic drainage on distant organs in intestinal ischemia-reperfusion injury in rats. METHODS: Forty-eight healthy Sprague-Dawley (SD) male rats (SPF grade) were randomly divided into 3 groups (16 rats in each group): normal diet group (N), enteral nutrition group (EN), enteral nutrition and ω-3PUFA group(PUFA group). Each group was divided into lymphatic drainage (I/R + D) group and no-drainage (I/R) group (n = 8). Each rats received gastrostomy. After given different nutrition for five days, the rats subjected to 60 min ischemia and 120 min reperfusion injury of the superior mesenteric artery. When the rats subjected to ischemia-reperfusion injury, drained intestinal lymph for 180 min in the I/R + D group. The serum level of alanine aminotransferase (ALT) and level of myeloperoxidase (MPO), nitric oxide (NO), total of nitric oxide synthase (tNOS), inducible nitric oxide synthase (iNOS) of lung were detected. The organ injury of lung and liver and the expression of high mobility group box 1(HMGB1, the endogenous ligand of TLR4) in these organs were investigated too. RESULTS: The serum level of ALT in PUFA I/R + D and I/R group and EN I/R + D group were significantly lower than that in normal diet I/R group: (46 ± 20), (53 ± 15), (46 ± 21) and (100 ± 60) U/L (P < 0.05), respectively. The level of MPO, NO, tNOS, iNOS in lung in the I/R + D group were significantly lower than those in I/R group (P < 0.05): MPO (0.73 ± 0.15):(0.85 ± 0.10) unit/grams wet slice; NO (0.72 ± 0.51):(1.79 ± 1.32) µmol/gprot; tNOS (0.46 ± 0.15):(0.78 ± 0.27) U/mgprot; iNOS (0.06 ± 0.04):(0.11 ± 0.07) U/mgprot, respectively. The level of tNOS in PUFA I/R group was significantly lower than that in normal diet I/R group: (0.56 ± 0.13):(0.78 ± 0.27) U/mgprot (P < 0.05). MPO, NO, INOS levels in PUFA group were reduced compared with those in EN and normal diet group. HE stained sections and HMGB1 immunohistochemistry results showed that the organ injury in I/R group was severer than that in I/R + D group. The expression of HMGB1 increased in I/R group. The organ injury and the expression of HMGB1 in PUFA group were less than that in the other two main groups. CONCLUSIONS: Lymphatic drainage can alleviate injury of distant organs after intestinal ischemia-reperfusion in rats. ω-3 polyunsaturated fatty acids can increase body resistance to injury and promote recovery.

Lymph duct ligation during ischemia/reperfusion prevents pulmonary dysfunction in a rat model with ω-3 polyunsaturated fatty acid and glutamine.

OBJECTIVE: The release of injurious factors into the mesenteric lymph from the ischemic intestine has been shown to contribute to lung injury and systemic inflammation after severe injury. We studied the effects of lung injury and systemic inflammatory reaction after intestinal ischemia/reperfusion and mesenteric lymph duct ligation with different nutritional statuses. METHODS: Rats (n = 72) were fed with a normal diet or received one of three diets (enteral nutrition, glutamine, or ω-3 polyunsaturated fatty acid) that were isocaloric and isonitrogenous. After 7 d, rats were subjected to 60 min of intestinal ischemia, ischemia plus mesenteric lymph duct ligation, or sham procedures. After 3 d of ischemia, the lymph nodes, lung, intestinal, liver, and blood were harvested and analyzed. RESULTS: In the different groups, lung injury, including levels of myeloperoxidase, nitric oxide, nitric oxide synthase, and the index of alveolar apoptosis, were partly prevented by mesenteric lymph duct ligation (P < 0.05). Likewise, the rats with ischemia/reperfusion, but not those with duct ligation plus ischemia/reperfusion, had a significant increase in intestinal permeability and decreased mucosal thickness. The serum cytokine and endotoxin concentrations were also lower in the lymph duct ligation groups, although there was no significant difference between lymph duct ligation and sham procedure. The lung and intestinal injuries were attenuated in the groups fed with glutamine and ω-3 polyunsaturated fatty acid. CONCLUSION: These results indicate that lymph duct ligation prevents lung injury, a systemic inflammation reaction, and gut-barrier dysfunction. Enteral glutamine and ω-3 polyunsaturated fatty acid modified the gut inflammation, prevented lung injury, and attenuated the systemic inflammation reaction.

[Integrated Chinese and Western medicine and clinical nutrition].

Nutritional support, an important measure for critical patients subject to monitoring, is widely used in clinical practice now. Nutrients have been used early in the Chinese medicine therapy. A number of Chinese medicine prescriptions show nutritional improvement and immune function enhancing effects on critical and/or postoperative patients, and some Chinese herbs are nutrient substances. Although the theoretical bases of Chinese medicine and Western medicine are different, they could work together in the clinical nutritional treatment to form a therapeutic measure with Chinese characteristics, which could promote the heritage and development of Chinese medicine. A discussion regarding the relationship between Chinese medicine drug-therapy, acupuncture and nutrition was also given.