Joint Detection of Serum Vitamin D,Body Mass Index,and Tumor Necrosis Factor Alpha for the Diagnosis of Crohn’s Disease

Objective Vitamin D(VD)deficiency was reported to contribute to the progression of Crohn’s disease(CD)and affect the prognosis of CD patients.This study investigated the role of serum VD,body mass index(BMI),and tumor necrosis factor alpha(TNF-α)in the diagnosis of Crohn’s disease.Methods CD patients(n=76)and healthy subjects(n=76)were enrolled between May 2019 and December 2020.The serum 25-hydroxyvitamin D[25(OH)D]levels,BMI,and TNF-αlevels,together with other biochemical parameters,were assessed before treatment.The diagnostic efficacy of the single and joint detection of serum 25(OH)D,BMI,and TNF-αwas determined using receiver operating characteristic(ROC)curves.Results The levels of 25(OH)D,BMI,and nutritional indicators,including hemoglobin,total protein,albumin,and high-density lipoprotein cholesterol,were much lower,and the TNF-αlevels were much higher in the CD patients than in the healthy subjects(P<0.05 for all).The areas under the ROC curve for the single detection of 25(OH)D,BMI,and TNF-αwere 0.887,0.896,and 0.838,respectively,with the optimal cutoff values being 20.64 ng/mL,19.77 kg/m^(2),and 6.85 fmol/mL,respectively.The diagnostic efficacy of the joint detection of 25(OH)D,BMI,and TNF-αwas the highest,with an area under the ROC curve of 0.988(95%CI:0.968–1.000).Conclusion The joint detection of 25(OH)D,TNF-α,and BMI showed high sensitivity,specificity,and accuracy in CD diagnosis;thus,it would be effective for the diagnosis of CD in clinical practice.

Tumor necrosis factor alpha receptor 1 deficiency in hepatocytes does not protect from non-alcoholic steatohepatitis, but attenuates insulin resistance in mice

BACKGROUND End-stage liver disease caused by non-alcoholic steatohepatitis(NASH)is the second leading indication for liver transplantation.To date,only moderately effective pharmacotherapies exist to treat NASH.Understanding the pathogenesis of NASH is therefore crucial for the development of new therapies.The inflammatory cytokine tumor necrosis factor alpha(TNF-α)is important for the progression of liver disease.TNF signaling via TNF receptor 1(TNFR1)has been hypothesized to be important for the development of NASH and hepatocellular carcinoma in whole-body knockout animal models.AIM To investigate the role of TNFR1 signaling in hepatocytes for steatohepatitis development in a mouse model of diet-induced NASH.METHODS NASH was induced by a western-style fast-food diet in mice deficient for TNFR1 in hepatocytes(TNFR1ΔHEP)and their wild-type littermates(TNFR1fl/fl).Glucose tolerance was assessed after 18 wk and insulin resistance after 19 wk of feeding.After 20 wk mice were assessed for features of NASH and the metabolic syndrome such as liver weight,liver steatosis,liver fibrosis and markers of liver inflammation.RESULTS Obesity,liver injury,inflammation,steatosis and fibrosis was not different between TNFR1ΔHEP and TNFR1fl/fl mice.However,Tnfr1 deficiency in hepatocytes protected against glucose intolerance and insulin resistance.CONCLUSION Our results indicate that deficiency of TNFR1 signaling in hepatocytes does not protect from diet-induced NASH.However,improved insulin resistance in this model strengthens the role of the liver in glucose homeostasis.

Moxibustion Treatment Restoring the Intestinal Epithelium Barrier in Rats with Crohn's Disease by Down-Regulating Tumor Necrosis Factor Alpha,Tumor Necrosis Factor Receptor 1,and Tumor Necrosis Factor Receptor 2

Objective： To investigate whether moxibustion regulates tumor necrosis factor alpha （TNF-α）, tumor necrosis factor receptor 1 （TNFR1）, and TNFR2 in the intestinal mucosa and to explore whether moxibustion could be used by means of this mechanism, to repair the intestinal epithelium barrier disruption in Crohn＇s disease （CD）. Methods： The CD rat models were established by tdnitrobenzene sulfonic acid （TNBs）, randomly divided into a model control （MC） group, an herb-partition moxibustion （HPM） group, a mild-warm moxibustion （MWM） group, and a salicylazosulfapyridine （SASP） group, and all were compared with a normal control （NC） group. The HPM and MWM groups were treated by moxibustion at Tianshu （ST25） and Qihai （RN6） for 14 days, and the SASP group obtained the SASP solution orally for the same pedod of time. The intestinal epithelium morphology and TNF-α, TNFR1, and TNFR2 contents were observed by the transmission electron microscopy and enzyme linked immunosorbent assay. Results： The sevedty of morphological changes in CD intestinal epithelium was obviously improved, and the levels of TNF-α, TNFR1, and TNFR2 in the intestinal mucosa all significantly decreased in the HPM and MWM groups. However, there were no significant differences between the HPM and MWM groups. Conclusion： The moxibustion therapies （HPM and MWM） could reduce intestinal inflammation and restore intestinal epithelium barrier disruption in CD, which might be due to down- regulating TNF- α, TNFR1, and TNFR2 in intestinal mucosa and improving intestinal epithelium morphology.

Role of tumor necrosis factor-alpha in the pathogenesis of atrial fibrillation

Background Tumor necrosis factor-alpha （TNF-a） is a pleiotropic proinflammatory cytokine and contributes to many kinds of cardiovascular diseases via its receptors （TNFR1/TNFR2）. We hypothesize that TNF-a plays a role in the pathogenesis of chronic atrial fibrillation （AF）. Methods Sixty-seven consecutive patients who were scheduled to have cardiac surgery were enrolled into the study. Thirty-one patients with rheumatic heart disease （RHD） and AF were enrolled as study group （AF group）. The sinus rhythm （SR） control groups consisted of 20 patients with RHD and 16 patients with coronary artery disease （CAD）. Peripheral blood sample was collected before the operation. About 5 mm3 left atrial tissue was disserted during the operation and was separated into three parts for Western blotting, real time polymerase chain reaction （RT-PCR） and immunohistochemistry （IHC） analysis. Results Compared with the controls （RHD SR and CAD SR）, the levels of TNF-α （（14.40±5.45） pg/ml vs. （4.20±3.19） pg/ml vs. （2.68±2.20） pg/ml, P=0.000） and its soluble receptor 1 （sTNFR1） （（1623.9±558.6） pg/ml vs. （1222.3±175.6） pg/ml vs. （1387.5±362.2） pg/ml, P=0.001） in plasma were higher in patients with AF. TNF-a level had positive correlation with the left atrial diameter （LAD） （r=0.642, P=0.000）. Western blotting analysis showed that the protein levels of TNF-α （0.618±0.236 vs. 0.234±0.178 vs. 0.180±0.103, P=0.000） were higher in patients with AF. The RT-PCR analysis results demonstrated that the mRNA expression of TNF-a （0.103±0.047 vs. 0.031±0.027 vs. 0.023±0.018, P=0.000） increased in patients with AF. IHC analysis displayed that, comparing to the SR, the expression of TNF-α （0.125±0.025 vs. 0.080±0.027 vs. 0.070±0.023, P=0.000） increased in the AF group. The protein level and mRNA expression of TNF-α also had positive correlation with left atrium diameter （LAD） （r=0.415, P=0.000 and r=0.499, P=0.000）. Conclusions The results revealed that TNF-α elevated in the plasma and left atrial tissue and had positive correlation with LAD in patients of chronic AF. TNF-α miaht involve in the pathogenesis of chronic AF.

Effect of cholinesterase inhibitor galanthamine on circulating tumor necrosis factor alpha in rats with lipopolysaccharideinduced peritonitis

Background The nervous system, through the vagus nerve and its neurotransmitter acetylcholine, can down-regulate the systemic inflammation in vivo, and recently, a role of brain cholinergic mechanisms in activating this cholinergic anti-inflammatory pathway has been indicated. Galanthamine is a cholinesterase inhibitor and one of the centrally acting cholinergic agents available in clinic. This study aimed to evaluate the effect of galanthamine on circulating tumor necrosis factor alpha （TNF-a） in rats with lipopolysaccharide-induced peritonitis and the possible role of the vagus nerve in the action of galanthamine. Methods Rat models of lipopolysaccharide-induced peritonitis and bilateral cervical vagotomy were produced. In the experiment 1, the rats were randomly divided into control group, peritonitis group, and peritonitis groups treated with three dosages of galanthamine. In the experiment 2, the rats were randomly divided into sham group, sham plus peritonitis group, sham plus peritonitis group treated with galanthamine, vagotomy plus peritonitis group, and vagotomy plus peritonitis group treated with galanthamine. The levels of plasma TNF-α were determined in every group. Results The level of circulating TNF-α was significantly increased in rats after intraperitoneal injection of endotoxin. Galanthamine treatment decreased the level of circulating TNF-α in rats with lipopolysaccharide-induced peritonitis, and there was significant difference compared with rats with lipopolysaccharide-induced peritonitis without treatment. The 3 mg/kg dosage of galanthamine had the most significant inhibition on circulating TNF-α level at all the three tested doses. Galanthamine obviously decreased the TNF-a level in rats with lipopolysaccharide-induced peritonitis with sham operation, but could not decrease the TNF-α level in rats with lipopolysaccharide-induced peritonitis with vagotomy. Conclusion Cholinesterase inhibitor galanthamine has an inhibitory effect on TNF-α release in rats with Iipopolysaccharide-induced peritonitis, and the vagus nerve plays a role in the process of the action of galanthamine.

Tumor necrosis factor alpha affect hydrocortisone expression in mice adrenal cortex cells mainly through tumor necrosis factor alpha-receptor 1

Background Tumor necrosis factor alpha （TNF-α） is important in promoting relative adrenal insufficiency （RAI） due to systemic inflammatory response syndrome （SIRS）. We identified the TNF-α receptor involved in the inhibition of adrenal corticotrophin （ACTH）-stimulated hydrocortisone release by studying the expression of TNF-α receptors in adrenal cortex Y1 cells and the effect of downregulating TNF receptors on ACTH-stimulated hydrocortisone release. Methods We used real-time PCR and immunocytochemistry to evaluate the expression of TNF receptors on Y1 cells. TNF-receptor 1 （TNF-R1） DNA fragments corresponding to the short hairpin RNA （shRNA）-sequences were synthesized and cloned into pcDNATM 6.2-GW/EmGFP expression vector. Knockdown efficiency of TNF-R1 expression was evaluated in miRNA transfected and mock-miRNA transfected Y1 cells by quantitative real-time PCR （Q-PCR）. Hydro- cortisone expression levels were determined in TNF-Rl-knockdown and control Y1 cells treated with TNF-α and ACTH. Results Mouse adrenal cortex Y1 cells were positive for type I TNF-R1, but not type II TNF-receptor （TNF-R2）. Blocking TNF-R1 expression resulted in loss of TNF-a-mediated inhibition of ACTH-stimulated hydrocortisone expression suggesting a role for the TNF-R1 related signaling pathway in ACTH-stimulated hydrocortisone synthesis. Conclusion The inhibitory effect of TNF-α on ACTH-stimulated hydrocortisone synthesis was mediated via TNF-R1 in adrenal cortex.

NO ASSOCIATION BETWEEN TUMOR NECROSIS FACTOR ALPHA AND OBSESSIVE COMPULSIVE DISORDER IN CHINESE HAN POPULATION

Objective To investigate association between tumor necrosis factor alpha (TNF-α) and obsessive compulsive disorder (OCD) in Chinese Han population.Methods Plasma concentrations of TNF-α were measured in 61 drug-free patients who fulfilled DSM-Ⅳ criteria for OCD and 93 healthy controls.TNF-α concentrations in blood were determined by enzyme-linked immunosorbent assay (ELISA).Two polymorphisms of TNF-α gene were investigated in the same patients and healthy controls:-308 G/A and-238 G/A.The allelic and genotypic frequencies of these polymorphisms were examined by using DNA sequencing method.Results Plasma levels of TNF-α did not differ between patients and controls (P=0.292).No significant results were observed for any of the alleles at the-308 G/A or-238 G/A polymorphism of the TNF-α gene.Finally a haplotype consisting of genotypes of these two markers was also examined.No association was observed for any haplotype (P=0.108).Conclusion No significant differences were observed between patients and controls in plasma levels of TNF-α.There is no association between the-308 G/A and-238 G/A TNF-α gene polymorphisms and OCD in our Chinese samples.However,the results need to be replicated in larger samples.

Comparative analysis of Nε-carboxymethyl-lysine and inflammatory markers in diabetic and non-diabetic coronary artery disease patients

BACKGROUND Coronary artery disease(CAD)is a major cause of death worldwide,and India contributes to about one-fifth of total CAD deaths.The development of CAD has been linked to the accumulation of Nε-carboxymethyl-lysine(CML)in heart muscle,which correlates with fibrosis.AIM To assess the impact of CML and inflammatory markers on the biochemical and cardiovascular characteristics of CAD patients with and without diabetes.METHODS We enrolled 200 consecutive CAD patients who were undergoing coronary angiography and categorized them into two groups based on their serum glycosylated hemoglobin(HbA1c)levels(group I:HbA1c≥6.5;group II:HbA1c<6.5).We analyzed the levels of lipoproteins,plasma HbA1c levels,CML,interleukin-6(IL-6),tumor necrosis factor alpha(TNF-α),and nitric oxide.RESULTS Group I (81 males and 19 females) patients had a mean age of 54.2 ± 10.2 years, with a mean diabetes duration of4.9 ± 2.2 years. Group II (89 males and 11 females) patients had a mean age of 53.2 ± 10.3 years. Group I had moresevere CAD, with a higher percentage of patients with single vessel disease and greater stenosis severity in the leftanterior descending coronary artery compared to group II. Group I also exhibited a larger left atrium diameter.Group I patients exhibited significantly higher levels of CML, TNF-α, and IL-6 and lower levels of nitric oxide ascompared with group II patients. Additionally, CML showed a significant positive correlation with IL-6 (r = 0.596,P = 0.001) and TNF-α (r = 0.337, P = 0.001) and a negative correlation with nitric oxide (r=-4.16, P = 0.001). Oddsratio analysis revealed that patients with CML in the third quartile (264.43-364.31 ng/mL) were significantlyassociated with diabetic CAD at unadjusted and adjusted levels with covariates.

Inhibition of tumor necrosis factor-α reduces alveolar septal cell apoptosis in passive smoking rats

Background Recent studies have revealed that lung cell apoptosis plays an important role in pathogenesis of cigarette-induced chronic obstructive pulmonary disease （COPD）. Tumor necrosis factor alpha （TNF-α） is one of the most important cytokines which are involved in COPD. This study aimed at investigating the influence of its inhibitor, recombinant human necrosis factor-alpha receptor II：lgG Fc fusion protein （rhTNFR：Fc） on alveolar septal cell apoptosis in passive smoking rats. Methods Forty-eight rats were randomly divided into a normal control group, a passive smoking group, an rhTNFR：Fc intervention group and a sham intervention group. The passive smoking rats were treated by exposure to cigarette smoking daily for 80 days. After smoking for one month the rhTNFR：Fc intervention group was treated with rhTNFR：Fc by subcutaneous injection, the sham intervention group injected subcutaneously with a neutral preparation （normal saline 0.1 ml, manicol 0.8 ml, cane sugar 0.2 mg, Tris 0.024 mg as a control. Lung function was determined and the levels of TNF-a in serum and broncho-alveolar lavage fluid （BALF） were measured with enzyme-linked immunosorbnent assay （ELISA）. Lung tissue sections stained by hematoxylin and eosin （HE） were observed for study of morphological alternations. Mean linear intercept （MLI） and mean alveolar numbers （MAN） were measured and the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） method was carried out to determine the percentage of positive cells and distribution of apoptotic cells. Results Increased MLI and decreased MAN were found in the passive smoking group compared with both the normal control group and the rhTNFR：Fc intervention group （P〈0.05）. Forced expiratory volume in 0.3 second （FEVo.3）/forced vital capacity （FVC） and peak expiratory flow （PEF） were lower in the passive smoking group than that in the normal control group （P〈0.05）. Compared with the sham intervention group, FEVo.3/FVC and PEF increased in the rhTNFR：Fc intervention group （P〈0.05）. The levels of TNF-α in serum were higher in the passive smoking group than that in the normal control group （P〈0.05） and rhTNFR：Fc intervention group （P〈0.05）. Significant differences were found between the levels of TNF-α in the serum of the rhTNFR：Fc intervention group and sham intervention group （P〈0.05）. The levels of TNF-α in BALF were higher in the passive smoking group than that in the normal control group （P〈0.05）, but no significant differences of TNF-α levels in BALF were found between the passive smoking group and rhTNFR：Fc intervention group. The number of TUNEL positive cells in alveolar septa was significantly increased in the passive smoking group as compared with the normal control group and the rhTNFR：Fc intervention group （P〈0.05）. Conclusion This study provides preliminary evidence that rhTNFR：Fc may interfere with TNF-α and reduce alveolar septal apoptosis in smoking rats.

Gut region-specific TNFR expression:TNFR2 is more affected than TNFR1 in duodenal myenteric ganglia of diabetic rats

BACKGROUND Cytokines are essential in autoimmune inflammatory processes that accompany type 1 diabetes.Tumor necrosis factor alpha plays a key role among others in modulating enteric neuroinflammation,however,it has a dual role in cell degeneration or survival depending on different TNFRs.In general,TNFR1 is believed to trigger apoptosis,while TNFR2 promotes cell regeneration.The importance of the neuronal microenvironment has been recently highlighted in gut region-specific diabetic enteric neuropathy,however,the expression and alterations of different TNFRs in the gastrointestinal tract has not been reported.AIM To investigate the TNFR1 and TNFR2 expression in myenteric ganglia and their environment in different intestinal segments of diabetic rats.METHODS Ten weeks after the onset of hyperglycemia,gut segments were taken from the duodenum,ileum and colon of streptozotocin-induced(60 mg/body weight kg i.p.)diabetic(n=17),insulin-treated diabetic(n=15)and sex-and age-matched control(n=15)rats.Myenteric plexus whole-mount preparations were prepared from different gut regions for TNFR1/HuCD or TNFR2/HuCD double-labeling fluorescent immunohistochemistry.TNFR1 and TNFR2 expression was evaluated by post-embedding immunogold electron microscopy on ultrathin sections of myenteric ganglia.TNFRs levels were measured by enzyme-linked immunosorbent assay in muscle/myenteric plexus-containing(MUSCLE-MP)tissue homogenates from different gut segments and experimental conditions.RESULTS A distinct region-dependent TNFRs expression was detected in controls.The density of TNFR1-labeling gold particles was lowest,while TNFR2 density was highest in duodenal ganglia and a decreased TNFRs expression from proximal to distal segments was observed in MUSCLE-MP homogenates.In diabetics,the TNFR2 density was only significantly altered in the duodenum with decrease in the ganglia(0.32±0.02 vs 0.45±0.04,P<0.05),while no significant changes in TNFR1 density was observed.In diabetic MUSCLE-MP homogenates,both TNFRs levels significantly decreased in the duodenum(TNFR1:4.06±0.65 vs 20.32±3.1,P<0.001;TNFR2:11.72±0.39 vs 15.91±1.04,P<0.01),which markedly influenced the TNFR2/TNFR1 proportion in both the ganglia and their muscular environment.Insulin treatment had controversial effects on TNFR expression.CONCLUSION Our findings show diabetes-related region-dependent changes in TNFR expression and suggest that TNFR2 is more affected than TNFR1 in myenteric ganglia in the duodenum of type 1 diabetic rats.

Effect of Two Chinese Medicinal Compounds,Blood-Activating and Water-Draining Medicine,on Tumor Necrosis Factor α and Nuclear Factor κ B Expressions in Rats with Intracerebral Hemorrhage

Objective：To investigate the effect of blood-activating Chinese medicinal compounds and waterdraining Chinese medicinal compounds on tumor necrosis factor alpha（TNF-α）and nuclear factor kappaB（NF-κB）expressions in rats with intracerebral hemorrhage（ICH）at the acute stage,and to monitor their therapeutic effect and mechanism of action on inflammation and cerebral edema.Methods：A rat model of cerebral hemorrhage was achieved by injecting autologous arterial blood into the caudate nucleus.A total of 168 rats were randomly divided into 4 groups：blood-activating medicine group（n=42）,water-draining medicine group（n=42）,sham operated group（n=42）,and the model group（n=42）.A series of brain samples were obtained at days 1,3 and 5 after ICH from rats in all groups.Protein expression levels of TNF-αand NF-κB were measured by immunohistochemical staining and gene expression levels of TNF-αand NF-κB were measured by real-time fluorescent PCR.Results：Compared to the sham operated group,protein expression levels of TNF-αand NF-κB in the model group significantly increased（P〈0.01）.Protein and gene expressions of TNF-αfrom the blood-activating medicine group and water-draining medicine group significantly decreased when compared to those in the model group（P〈0.05）.Meanwhile,compared to the model group,the expression of NF-κB in the blood-activating medicine group significantly decreased（P〈0.05）,while expression of NF-κB in the water-draining medicine group did not differ（P〉0.05）.Conclusions：Blood-activating Chinese medicinal compounds and water-draining Chinese medicinal compounds can alleviate inflammation of peripheral tissue and cerebral edema.However,the blood-activating Chinese medicinal compounds were more effective than the water-draining Chinese medicinal compounds.The possible effective mechanism may be by means of inhibiting the activation of NF-κB so as to suppress the transcription of target genes including gene expression of TNF-α.

Altered expression of miR-125a and dysregulated cytokines in systemic lupus erythematosus: Unveiling diagnostic and prognostic markers

BACKGROUND Systemic lupus erythematosus(SLE)is a chronic autoimmune disorder impacting multiple organs,influenced by genetic factors,especially those related to the immune system.However,there is a need for new biomarkers in SLE.MicroRNA-125a(miR-125a)levels are decreased in T cells,B cells,and dendritic cells of SLE patients.MiR-125a plays a regulatory role in controlling the levels of tumor necrosis factor-alpha(TNF-α)and interleukin 12(IL-12),which are crucial pro-inflammatory cytokines in SLE pathogenesis.AIM To assess the levels of miR-125a,IL-12,and TNF-αin SLE patients’plasma,evaluating their diagnostic and prognostic value.METHODS The study included 100 healthy individuals,50 newly diagnosed(ND),and 50 SLE patients undergoing treatment.The patients were monitored for a duration of 24 wk to observe and record instances of relapses.MiR-125a expression was measured using real-time reverse transcription polymerase chain reaction,while ELISA kits were used to assess IL-12 and TNF-αproduction.RESULTS The results showed significantly reduced miR-125a expression in SLE patients compared to healthy individuals,with the lowest levels in ND patients.TNF-αand IL-12 expression levels were significantly elevated in SLE patients,especially in the early stages of the disease.Receiver operating characteristic curve analyses,and Cox-Mantel Log-rank tests indicated miR-125a,TNF-α,and IL-12 as proper diagnostic biomarkers for SLE.A negative correlation was found between plasma miR-125a expression and IL-12/TNF-αlevels in SLE patients.CONCLUSION Decreased miR-125a levels may be involved in the development of SLE,while elevated levels of IL-12 and TNF-αcontribute to immune dysregulation.These findings offer new diagnostic and prognostic markers for SLE.Moreover,the negative correlation observed suggests an interaction between miR-125a,TNF-α,and IL-12.Further research is necessary to uncover the underlying mechanisms that govern these relationships.

Changes of gut bacteria and immune parameters in liver transplant recipients

BACKGROUND: Liver transplantation is one of the most effective therapeutic options for patients with end-stage liver diseases, and gut microbiota is actively involved in potential infections in pretransplant and posttransplant patients. However, the diversity of gut microbiota and its relationship with the immune parameter of liver transplantation recipients are not well understood. METHODS: We collected fresh feces and blood samples from 190 participants in China from November 2004 to May 2008, including 28 healthy volunteers, 51 cirrhotic patients and 111 liver-transplanted patients. Six interesting gut bacteria, plasma endotoxin, serum cytokines (i.e., tumor necrosis factor alpha and interleukin-6) and fecal secretory IgA (SIgA) were investigated by real-time quantitative PCR, chromogenic limulus amoebocyte assay, sandwich-type enzyme-linked immunosorbent assay and radioimmunoassay, respectively. RESULTS: All Eubacteria, Bifidobacterium spp., Faecalibacterium prausnitzii and Lactobacillus spp. were significantly lower in the liver transplantation recipients while Enterobacteriaceae and Enterococcus spp. were significantly higher (P<0.05). Except for Enterococcus spp., other bacteria showed a tendency to restore to normal level along with the time after liver transplantation. Plasma endotoxin, interleukin-6 and fecal SIgA in cirrhotic patients increased significantly, but not in liver transplantation recipients. Plasma endotoxin and interleukin-6 were negatively correlated with all Eubacteria and the Bacteroides-Prevotella group, while tumor necrosis factor alpha was not significantly correlated with these six gut bacteria in cirrhotic patients.CONCLUSIONS: Our study demonstrates that abundant gut bacteria were altered significantly in both cirrhotic and liver transplantation patients, while plasma endotoxin and interleukin-6 increased remarkably in cirrhotic patients, showing significant correlations with gut microbiota. Interestingly, our data show a tendency for these gut bacteria to restore to normal levels in liver transplantation recipients.

Anti-inflammatory properties of lipoxin A4 protect against diabetes mellitus complicated by focal cerebral ischemia/reperfusion injury

Lipoxin A4 can alleviate cerebral ischemia/reperfusion injury by reducing the inflammatory reaction,but it is currently unclear whether it has a protective effect on diabetes mellitus complicated by focal cerebral ischemia/reperfusion injury.In this study,we established rat models of diabetes mellitus using an intraperitoneal injection of streptozotocin.We then induced focal cerebral ischemia/reperfusion injury by occlusion of the middle cerebral artery for 2 hours and reperfusion for 24 hours.After administration of lipoxin A4 via the lateral ventricle,infarction volume was reduced,the expression levels of pro-inflammatory factors tumor necrosis factor alpha and nuclear factor-kappa B in the cerebral cortex were decreased,and neurological functioning was improved.These findings suggest that lipoxin A4 has strong neuroprotective effects in diabetes mellitus complicated by focal cerebral ischemia/reperfusion injury and that the underlying mechanism is related to the anti-inflammatory action of lipoxin A4.

Protective Effect of Diallyl Trisulfide on Liver in Rats with Sepsis and the Mechanism

The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-operated group (group S, n=8), sepsis model group (group C, n=24), diallyl trisulfide (DATS)-treated group (group D, n=24). Animals in groups C and D were further divided into three subgroups according to different observation time points, with 8 rats in each sub-group.Rats in group D and C were intravenously injected with normal saline or DATS respectively at a dose of 20 mg/kg after the establishment of sepsis model. Eight rats in groups C and D were sacrificed at 3, 6 and 24 h post-CLP and their livers were harvested for detection of interleukin (IL)-1 receptor associated kinase-4 (IRAK-4), nuclear factor-κB (NF-κB), c-fos, c-jun, malondialdehydethhe (MDA) and superoxide dismutase (SOD), tumor necrosis factor alpha (TNF-α) and for pathological examination. The results showed that the levels of serum IRAK-4, NF-κB and TNF-α in hepatic tissues were higher in group C than group S (control group) (P<0.05). After DATS treatment, the levels of IRAK-4 and NF-κB in the hepatic tissues and serum TNF-α in group D were lower than those in group C (P<0.05). The levels of c-fos and c-jun and MDA in the hepatic tissues were higher in group C than in group S (P<0.05). After DATS treatment, the levels of c-fos and c-jun and MDA in the hepatic tissues were significantly lower in group D than in group C (P<0.05). When compared with group S group, concentration of SOD in the hepatic tissues in group C was significantly lower (P<0.05). After DATS treatment, the concentration of SOD in the hepatic tissues was higher in group D than in group C (P<0.05). These findings suggested that treatment with DATS could ameliorate sepsis-induced liver injury in rats. The protective effect might be related to its ability to inhibit the signal pathway of IRAK-4 and NF-κB, thereby decreasing the production of oxygen free radicals and down-regulating the expression of c-fos and c-jun.

Active tuberculosis in inflammatory bowel disease patients under treatment from an endemic area in Latin America

BACKGROUND There has been an increase in cases of inflammatory bowel disease(IBD)in recent years.There is also greater access and availability of immunosuppressive and biological agents,which increase the risk of opportunistic infection despite improving the quality of life and promoting mucosal healing.Tuberculosis(TB)remains a public health problem,and it has a high incidence in several countries.Therefore,knowledge of the risk of developing TB in patients with IBD is important.AIM To evaluate the risk of active TB in patients with IBD under treatment from an endemic area in Latin America.METHODS A standard questionnaire included demographic variables,clinical aspects of IBD disease,history of active TB during treatment,active TB characteristics and evolution,initial screening and results and time from the start of anti-tumor necrosis factor alpha(TNFα)to TB development.RESULTS Azathioprine,anti-TNFα and the combination of these two drugs were associated with a higher risk of active TB incidence.The TNFα blockers increased the relative risk of developing active TB compared to other treatments.All four multivariable models showed that the use of TNFα blockers alone or in combination with azathioprine was an important risk factor for the incidence of active TB.After adjustment for sex,age,type of IBD and latent TB,anti-TNFα with azathioprine increased the relative risk to 17.8 times more than conventional treatment.Late TB,which was diagnosed 3 mo after the start of anti-TNFα,was the most frequent.CONCLUSION Treatment with anti-TNFα increased the risk of active TB in IBD patients from an endemic area in Latin America.This risk was increased when anti-TNFα was combined with azathioprine.The time from the beginning of the treatment to the active TB diagnosis suggests a new TB infection.

Update on biologic therapies for juvenile idiopathic arthritis-associated uveitis

Juvenile idiopathic arthritis(JIA)is the most common rheumatic disease of childhood,and juvenile idiopathic associated uveitis(JIA-U)is the most frequently noted extra-articular manifestation.JIA-U can present asymptomatically and lead to ocular complications,so regular screening and monitoring are needed to prevent potentially sight-threatening sequelae.Topical glucocorticoids such as prednisolone acetate are usually the first line of treatment for anterior uveitis associated with JIA-U,but long-term use may be associated with cataract,ocular hypertension and glaucoma.Disease modifying anti-rheumatic drugs(DMARDs)such as methotrexate allow tapering of the corticosteroids to prevent long-term complications.Biologic therapies have been increasingly used as targeted therapies for JIA-U,particularly monoclonal antibodies targeting the proinflammatory cytokine TNF-αsuch as adalimumab and infliximab.One recent,multicenter,prospective,randomized clinical trial provided evidence of the efficacy of adalimumab with methotrexate for JIA-U compared to methotrexate alone.Another clinical trial studying the interleukin-6 inhibitor tocilizumab for JIA-U showed promise in tapering topical corticosteroids.Additionally,JAK inhibitors are emerging biologic therapies for JIA-U in patients refractory to TNF-αinhibitors,with a clinical trial assessing the efficacy of baricitinib for JIA-U underway.While clinical trials on these novel biologics are limited,further investigation of these agents may provide additional therapeutic options for JIA-U.

Relationship between peritoneal macrophages and inflammatory reaction in a rat model of severe acute pancreatitis

Objective To investigate the relationship between peritoneal macrophages(PMAs)and inflammatory reaction in a rat model of severe acute pancreatitis(SAP).Methods Sprague-Dawley rats were randomly divided into control group and SAP group.To induce SAP in rats,40 g/L sodium taurocholate(0.1 mL/100 g)was injected into the pancreatic duct through retrograde exposure of pancreatic bile duct in hepatic porta.One-third of rats were sacrificed at 3,6 or 12 h after modeling.PMAs were extracted,and incubated for 24 h in a humidified 5% carbon dioxide incubator.The expressions of tumor necrosis factor alpha(TNF-α)and interleukin-1β(IL-1β)mRNA in PMAs were measured by semi-quantitative RT-PCR.The levels of TNF-α and IL-1β in culture medium and serum were evaluated.The histological changes of pancreas were examined.Results The expressions of TNF-α mRNA and IL-1β mRNA in PMAs were significantly higher in SAP group than in control group at each time point(P<0.01).The concentrations of TNF-α and IL-1β in culture medium and serum were significantly elevated in SAP group compared with control group(P<0.01).The histological analysis of pancreas indicated that the damage was more severe in SAP group than in control group(P<0.01).Conclusion PMAs secrete cytokines into pancreatitis-associated ascitic fluid,and this study demonstrates a correlation between SAP and the activation of PMAs.

Therapeutic Mechanism of Santeng Dingtong Recipe (三藤定痛方) on Monosodium Urate Crystal-Induced Rabbit Arthritis

Objective: To study the therapeutic mechanism of Santeng Dingtong recipe (STDT) on monosodium urate crystal (MSU) induced rabbit arthritis Methods: Forty-two rabbits were randomly divided into six groups, 7 in each group. Group 1 received 0.9% saline 2. 5 ml/kg per day by gastrogavage (ig) for 10 days; Group 2, 3 and 4 received STDT 0.125 g/kg, 1.0 g/kg and 8.0 g/kg per day respectively by ig for 10 days; Group 5 received colchicine 4. 5 mg/kg per day by ig for 4 days; and Group 6 was untreated. MSU crystals 10 mg /500ul containing polymyxin B 10 u/ml was injected into the knee joints of Group 1-5 to make rabbit arthritis models. Leukocytes in synovial lavage fluids was then counted and differentiated; pathological injury of synovial membranes was observed under HE staining; interleukin-1 beta (IL-1B), tumor necrosis factor alpha (TNFa) and leukotriene B4 (LTB4) content in synovial lavage fluids were determined by ELISA. Results: MSU caused a rapid leukocyte infiltration and increased production of IL-1B, TNFa and LTB4 2 hrs after intra-articular injection. STDT inhibited neutrophil infiltration in synovial fluids dose-dependently, protected the synovial membrane against pathological injury and reduced the production of IL-1B, TNFa and LTB4; while colchicine did not decrease the level of TNFa, but significantly inhibited neutrophil infiltration in synovial fluid and reduced the production of IL-11B and LTB4. Conclusion: STDT exerts an anti-inflammatory effect in rabbit model of acute MSU arthritis, its mechanism being probably due to the decrease of XL-1B, TNFa and LTB4 synthesis.

Neuroprotective role of Batroxobin in cardiopulmonary resuscitation rabbits

BACKGROUND： Batroxobin has been found to have protective effect on cerebral ischemia-reperfusion, and cardiopulmonary resuscitation （CPR） is the common cause of global brain ischemia-reperfusion. OBJECTIVE： To observe the effect of Batroxobin on the morphological results of cerebral cortex and hippocampus in rabbit models of CPR, and the changes of serum concentration of tumor necrosis factor alpha （TNF- α ） after CPR. DESIGN： A randomized controlled observation. SETTING： Laboratory of the Department of Bums, Changhai Hospital affiliated to the Second Military Medical University of Chinese PLA. MATERIALS： Thirty healthy New Zealand rabbits of 2.5 - 3.0 kg, either male or female, were used. Kits for TNF- α determination were provided by LIFEKEY BioMeditech Company （USA）. METHODS： The experiments were carried out in the laboratory of Department of Bums, Changhai Hospital from February 2001 to January 2002. The 32 rabbits were randomly divided into sham-operated group （n=8）, conventional resuscitation group （n=12） and Batroxobin-treated group （n=12）. The animals in the conventional resuscitation group and Batroxobin-treated group were anesthetized, then induced into modified Pittsburg＇s model of mechanical ventricular fibrillation. Sham-operated group was discharged on the chest wall, which did not cause ventricular fibrillation. Conventional resuscitation group and Batroxobin-treated group were exposed to 6 minutes of cardiac arrest induced by ventricular fibrillation, then the resuscitation began. A dosage of 0.3 Bu/kg of Batroxobin was administered to the rabbits in the Batroxobin-treated group at the beginning of resuscitation. Blood sample was collected at 4 and 12 hours after CPR to determine the concentration of TNF- α in serum. After the second blood collection, brain tissue was taken out immediately, and the forms of nerve cells in cerebral cortex and hippocampal CAl region were observed under light microscope. MAIN OUTCOME MEASURES： ① TNF- α concentration in serum at 4 and 12 hours after CPR; ② Forms of nerve cells in cerebral cortex and hippocampal CAI region at 12 hours after CPR. RESULTS： All the 31 New Zealand rabbits were involved in the analysis of results. ①TNF-α concentration in serum： At 4 hours after CPR, the TNF-α concentrations in serum in the conventional resuscitation group and Batroxobin-treated group [（5.947±2.366）, （5.122±2.521） ng/L] were significantly higher than that in the sham-operated group [（2.604±1.623） ng/L, P 〈 0.05]. At 12 hours after CPR, the TNF- α concentration in serum in the conventional resuscitation group was （7.770±3.121） ng/L, it was significantly higher than that at 4 hours （P 〈 0.05）, also significantly higher than that in the Batroxobin-treated group [（5.425±2.280） ng/L, P 〈 0.05]. ② Forms of nerve cells： In the sham-operated group, no abnormality was found in the hippocampal CAI region and cerebral cortex. In the conventional resuscitation group, the pyramidal cells in hippocampal CAI region were lined up in disorders, and edema, puff, vacuolization, nucleus concentration and anachromasis were also observed appeared; Edema of nerve cells, vacuole, pyknosis appeared in cerebral cortex; microthrombosis appeared in some blood capillaries. As compared with the conventional resuscitation group, cellular edema was relieved and pyknosis of nerve cells were obviously reduced, and no microthrombosis was found in hippocampal CAI region and cerebral cortex in the Batroxobin-treated group. CONCLUSION： Batroxobin have neuroprotective effect on CPR rabbits, and may inhibit the excessive increase of TNF- α concentration in serum.