[Effects of Hydrogen Sulfide on Inflammatory Factors and Mitochondrial Energy Metabolic Disorders After Reperfusion Injury in Rats].

Objective To explore the effect of hydrogen sulfide on inflammatory factors and energy metabolism of mitochondria after limbs reperfusion injury in rats. Methods Sixty rats were divided into three groups:sham operation group,control group(ischemia-reperfusion injury + saline group),and experimental group(ischemia-reperfusion injury + H2S group).Wistar rat models of limb ischemia-reperfusion injury were established.Skeletal muscle samples were collected to determine the levels of necrosis decomposition products [including myoglobin(MB),lipoprotein complex(LPC)and lipid peroxide(LPO)];blood samples were collected to determine the levels of interleukin(IL)-1,IL-6 and tumor necrosis factor-α(TNF-α);mitochondria were extracted for mitochondrial transmembrane potential measurement and ATP content detection.Statistical analysis was made on the test results. Results After ischemia reperfusion injury,the levels of MB,LPO,and LPC in skeletal muscle,liver,lung and renal tissues of the control group were significantly increased(MB:Pskeletal muscle =0.003,Pliver =0.001,Plung =0.001,Pkidney =0.001;LPO:Pskeletal muscle =0.001,Pliver =0.001,Plung =0.001,Pkidney =0.002;LPC:Pskeletal muscle =0.000,Pliver =0.002,Plung =0.002,Pkidney =0.003),and hydrogen sulfide treatment during ischemia reperfusion significantly inhibited the production of MB,LPO,and LPC(MB:Pskeletal muscle =0.021,Pliver =0.036,Plung =0.005;LPO:Pskeletal muscle =0.003,Pliver =0.008,Plung =0.010,Pkidney =0.015;LPC:Pskeletal muscle =0.002,Pliver =0.026,P lung =0.007,P kidney =0.006).Ischemia/reperfusion of lower extremity in rats resulted in increased levels of IL-1,IL-6,and TNF-α in the serum of rats,and the levels of IL-1,IL-6,and TNF-increased over time,with statistically significant differences in IL-1,IL-6,and TNF-α among groups at 3 h(IL-1:P3 h =0.019,P6 h =0.011,P9 h =0.009,$P_{12_{h}}$=0.008,and P15 h =0.002;IL-6:P3 h =0.026,P6 h =0.009,P9 h =0.002, $P_{12_{h}}$=0.002,P15 h =0.003;TNF-α:P3 h =0.002,P6 h =0.002,P9 h =0.005,$P_{12_{h}}$=0.002,P15 h =0.003).The levels of IL-1,IL-6,and TNF-α in serum were significantly inhibited during ischemia reperfusion(IL-1:P3 h =0.035,P6 h =0.039,P9 h =0.012,$P_{12_{h}}$=0.005,P15 h =0.006;IL-6:P3 h =0.042,P6 h =0.025,P9 h =0.023,$P_{12_{h}}$=0.006,P15 h =0.005;TNF-α:P3 h =0.005,P6 h =0.003,P9 h =0.022,$P_{12_{h}}$=0.005,P15 h =0.005),and such inhibitory effects became even more obvious over time.After limb ischemia and reperfusion in the control group,the mitochondrial transmembrane potential of skeletal muscle cells significantly decreased compared with that of the sham group(t=6.698;P=0.001).After hydrogen sulfide treatment,the mitochondrial membrane potential energy of the experimental group was significantly higher than that of the control group(t=7.507,P = 0.000).The ATP level in the mitochondria of ischemia reperfusion rats in the control group was significantly lower than that in the sham group(t=7.526,P = 0.000).The content of mitochondrial ATP in the experimental group was significantly higher than that in the control group after hydrogen sulfide treatment(t=8.604,P = 0.000). Conclusions Hydrogen sulfide can alleviate the injury of skeletal muscle and distal organs after limb ischemia-reperfusion and reduce local inflammatory reaction.In addition,it is valuable in alleviating mitochondrial transmembrane potential and energy metabolism disorders during reperfusion injury.

[Detection of immunity and antioxidant indexes in dairy cows infected with Cryptosporidium].

OBJECTIVE: To detect the immune status and antioxidant system indexes of cows infected with Cryptosporidium. METHODS: Fecal samples of 325 dairy cows were collected at a farm in Anhui and examined by floating saturated solution. 7 positive cows and 7 negative cows from the farm were selected as infection group and non-infection group, respectively. Blood samples were taken from cow's jugular vein before feeding in the morning. 19 indexes of total protein (TP), albumin (ALB), IgG, IgM, IgA, phagocytic rate of white blood cells, T lymphocyte transformation rate, IL-2, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), NO, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glucose (GLU), triglyceride (TG), Cl-, and Ca2+ were tested, respectively. RESULTS: The infection rate of 325 cows was 31.7% (103/325). The Cryptosporidium was identified as C. andersoni according to the morphology and size of oocysts. Compared with the non-infection group, there was no significant difference in the concentration of TP, ALB, IgM, IgA, GSH-Px, ALT, AST, ALP and Cl- (P > 0.05). The concentration of MDA and NO in the infection group increased by 59.9% and 28.1% (P < 0.05 or 0.01), and that of IgG, SOD, GLU, TG, Ca2+, IL-2 and the activities of T lymphocyte transformation rate, phagocytic rate of white blood cells decreased by 32.9%, 11.1%, 18.6%, 78.9%, 14.5%, 7.0%, 22.0%, and 20.2%, respectively (P < 0.05). CONCLUSION: The change of antioxidant and immune indexes shows that the capability of eliminating free radicals and the immune function have decreased in the Cryptosporidium andersoni-infected cows.

Clinical analysis of the risk factors of slow coronary flow.

Slow coronary flow (SCF) phenomenon is a coronary microvascular disorder characterized by the delayed passage of contrast in the absence of obstructive epicardial coronary disease, and is an important clinical entity because it may be the cause of precordial pain when the body is at rest and/or during exercise. Although clinical and pathological features of SCF have been previously described, its etiopathogenesis remains unclear. The present study aims to investigate the risk factors of slow coronary flow, in order to provide the foundation for further exploration of potential mechanisms of SCF. A total of 47 consecutive patients with documented slow coronary flow, and 33 patients with normal coronary flow--as defined by TIMI frame count (TFC)--were recruited for this study. Clinical information was collected, and biochemical indicators including high-sensitivity C-reactive protein (hs-CRP), and a marker of systemic inflammation were detected. Logistic regression analysis was performed for statistical analysis. SCF patients had a higher level of serum uric acid (323.2 ± 79.3 vs. 282.8 ± 82.4 µmol/l, p = 0.03), 2-h postprandial blood glucose (8.6 ± 2.7 vs. 7.5 ± 1.8 mmol/l, p = 0.04), platelet count (165.9 ± 51.6 × 10(3) vs. 127.0 ± 32.0 × 10(3) cells/µl, p = 0.0003) and hs-CRP (3.4 ± 0.8 vs. 2.0 ± 0.9 mg/l, p < 0.0001) than those of patients in the control group. No marked differences in other variables were observed between the two groups. Logistic regression showed serum uric acid level (χ(2) = 3.84, ß = 0.007, p = 0.049), 2-h postprandial blood glucose (χ(2) = 5.02, ß = 0.277, p = 0.025) and blood platelet count (χ(2) = 12.16, ß = 0.026, p = 0.001) were independent predictors of SCF. When hs-CRP was included in the multivariate model, hs-CRP (χ(2) = 21.19, ß = 1.90, p < 0.0001) was the only independent predictor of SCF. These findings suggested that an elevation of serum uric acid level, 2-h postprandial blood glucose, and blood platelet count might be the causes of SCF, and inflammation was likely to be implicated in the causal pathway leading to SCF.

Clinical effect of vestibular rehabilitation on benign paroxysmal positional vertigo: A protocol for systematic review.

BACKGROUND: This study will evaluate the clinical effect of vestibular rehabilitation (VR) on benign paroxysmal positional vertigo (BPPV). METHODS: In this study, we will identify relevant trials on the topic published in MEDLINE, EBASE, Web of Science, Cochrane Library, Scopus, CINAHL, CBM, and CNKI from inception to the present. We will also search conference proceedings, thesis/dissertation, ongoing trials in clinical trial registry, and reference lists of included studies. Two researchers will independently carry out record selection, data extraction, and study quality assessment, respectively. Any disagreement will be arbitrated and solved with the help of a third researcher. If necessary, we will conduct random-effects meta-analysis to pool the effect estimates of included trials determined to be acceptable heterogeneity. RESULTS: We will summarize the latest evidence to assess the effect of VR for the treatment of patients with BPPV. CONCLUSION: The findings of this study will help determine whether or not VR is effective in treating BPPV. OSF REGISTRATION: osf.io/k83y5.

Dynamic changes in HCN2, HCN4, KCNE1, and KCNE2 expression in ventricular cells from acute myocardial infarction rat hearts.

AIMS: To investigate dynamic changes in the expression of HCN2, HCN4, as well as KCNE1, and KCNE2 mRNA and protein levels in ventricular cells from acute myocardial infarction (AMI) rat hearts. MAIN METHODS: An AMI model was induced by ligating the left anterior descending coronary artery (LAD) of Sprague-Dawley rats. The rats were randomly divided into four experimental groups: 24-hour (24h) post-AMI, 1-week (1w) post-AMI, 2-week (2w) post-AMI, and 4-week (4w) post-AMI; sham-operated control rat groups were established in parallel for each time point. HCN2, HCN4, KCNE1, and KCNE2 mRNA and protein levels were measured by reverse transcription-polymerase chain reaction (RT-PCR) and by immunohistochemistry and Western blot, respectively. KEY FINDINGS: Ventricular arrhythmias occurred in all the post-AMI groups, particularly in the 1w and 2w post-AMI groups. Although HCN2, HCN4, KCNE1, and KCNE2 genes were expressed in the left ventricular myocardium of sham-operated control rats, their expression increased in rat ischemic left ventricular myocardium, with dynamic changes in expression observed 4 weeks after AMI. HCN2, HCN4, and KCNE2 protein levels were highest at 1w and KCNE2 protein levels peaked at 2w post-AMI. SIGNIFICANCE: The expression of the HCN2, HCN4, as well as KCNE1, and KCNE2 genes in ventricular cells from AMI rat hearts underwent dynamic changes, reaching peak levels at 1 or 2weeks post-AMI. The increased expression maybe related to ventricular arrhythmogenesis after AMI.

The individual and combined effects of deoxynivalenol and aflatoxin B1on primary hepatocytes of Cyprinus carpio.

Aflatoxin B(1) (AFB(1)) and deoxynivalenol (DON) are important food-borne mycotoxins that have been implicated in animal and human health. In this study, individual and combinative effects of AFB(1) and DON were tested in primary hepatocytes of Cyprinus carpio. The results indicated that the combinative effects of AFB(1) and DON (0.01 µg/mL AFB(1) and 0.25 µg/mL DON; 0.02 µg/mL AFB(1) and 0.25 µg/mL DON; 0.02 µg/mL AFB(1) and 0.5 µg/mL DON) were higher than that of individual mycotoxin (P < 0.05). The activity of AST, ALT and LDH in cell supernatant was higher than that of control group (P < 0.05) when the mycotoxins were exposed to primary hepatocytes for 4 h. The decreased cell number was observed in tested group by inverted light microscopy. The mitochondrial swelling, endoplasmic reticulum dilation and a lot of lipid droplets were observed in primary hepatocytes by transmission electron microscope. Therefore, this combination was classified as an additive response of the two mycotoxins.

Deoxynivalenol Induces Intestinal Damage and Inflammatory Response through the Nuclear Factor-κB Signaling Pathway in Piglets.

Deoxynivalenol (DON) is highly toxic to animals and humans, but pigs are most sensitive to it. The porcine mucosal injury related mechanism of DON is not yet fully clarified. Here, we investigated DON-induced injury in the intestinal tissues of piglet. Thirty weanling piglets [(Duroc × Landrace) × Yorkshire] were randomly divided into three groups according to single factor experimental design (10 piglets each group). Piglets were fed a basal diet in the control group, while low and high dose groups were fed a DON diet (1300 and 2200 µg/kg, respectively) for 60 days. Scanning electron microscopy results indicated that the ultrastructure of intestinal epithelial cells in the DON-treated group was damaged. The distribution and optical density (OD) values of zonula occludens 1 (ZO-1) protein in the intestinal tissues of DON-treated groups were decreased. At higher DON dosage, interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α mRNA levels were elevated in the intestinal tissues. The mRNA and protein levels of NF-κB p65, IκB-α, IKKα/ß, iNOS, and COX-2 in the small intestinal mucosa were abnormally altered with an increase in DON concentration. These results indicate that DON can persuade intestinal damage and inflammatory responses in piglets via the nuclear factor-κB signaling pathway.

A sensitive immunochromatographic assay using colloidal gold-antibody probe for rapid detection of fumonisin B1 in corn.

A sensitive immunochromatographic assay (ICA) using a colloidal gold-antibody probe for the rapid detection of fumonisin B1 (FB1) in corn samples was developed. The colour density of the test line correlated with the concentration of FB1 in the range 2-40 ng ml(-1) by the assay, and the detection limit for FB1 was 2 ng ml(-1). The linear range for FB1 was 50-1000 µg kg(-1), and the visual limit detection of the test was 1000 µg kg(-1) in corn samples. The ICA to detect FB1 is sensitive, specific and rapid. Specific anti-FB1 monoclonal antibody (mAb) and FB1-ovalbumin (FB1-OVA) conjugate antigen were prepared. FB1 mAb, labelled with colloidal gold, was used as the probe on the immunochromatographic strip. FB1-OVA and goat-anti-mouse IgG were coated onto a nitrocellulose (NC) membrane as test lines and control lines, respectively. FB1 in samples will competitively combines the FB1 mAb with the FB1-OVA in an NC membrane and the results are directly observed by the colour of the detection and quality control lines. The concentrations of FB1 mAb labelled with colloidal gold, detecting antigen and goat anti-mouse IgG, were optimised. The results indicate that the test strip is specific for FB1, with no cross-reactivity to other toxins. The strip assay for FB1 was simple, only needing one step without complicated assay performance and expensive equipment, and the total time for visual evaluation was less than 10 min. A survey of 24 corn samples from Hefei, China, was performed with the test strip and HPLC, and the detection results showed that the developed ICA and the HPLC were in excellent agreement. Hence, the developed ICA can be used as a method for rapid detection of FB1 in corn samples.

Effect of initial fluid resuscitation on subsequent treatment in uncontrolled hemorrhagic shock in rats.

SUMMARY: Using a modified uncontrolled hemorrhage shock model with massive splenic and vascular injury, we evaluated outcome and tissue oxidation injury with different resuscitation interventions during prehospital and hospital phases. The aim of our study was to explore the effect of initial fluid resuscitation on subsequent treatment of uncontrolled hemorrhagic shock in rats. Uncontrolled hemorrhagic shock was produced in 114 Wistar rat by sharp transection both of the splenic parenchyma at one location between the major branches of the splenic artery into the spleen and of one of the major branches of the splenic artery. Experimental design consisted of three phases: a "prehospital phase" (resuscitation with balanced saline to a mean arterial pressure (MAP) of 40, 50, 60, 80, and 100 mmHg, respectively, when MAP reached 30 mmHg), followed by a "hospital phase" (120 min, including control of hemorrhage and resuscitation with balanced saline and whole blood (2:1) or balanced saline alone to a MAP >80 mmHg), and a 240-min observation phase. Blood loss, infused volume, hematocrit, and survival rate were recorded. At the end of the experiment, survivors were sacrificed, and the lung, kidney, and distal ileum were harvested for determination of malondialdehyde (MDA) content and total antioxidative capacity (T-AOC). All rats that were resuscitated to a MAP >80 mmHg in the prehospital phase and received balanced saline alone in the hospital phase died, whereas those that had been resuscitated to a MAP of 40 or 50 mmHg during the prehospital phase and then resuscitated with balanced saline and whole blood in the hospital phase survived throughout the experiment. The animals whose MAP was kept higher than 80 mmHg had significantly higher MDA content and lower T-AOC than those whose MAP was maintained 40, 50, or 60 mmHg during the prehospital phase. In the hospital phase, resuscitation with balanced saline and whole blood not only relieved tissue damage but also improved the survival, as indicated by 44.4% survival rate in the rats that resuscitated to a MAP of 80 or 100 mmHg in the prehospital phase. These results suggested that in our uncontrolled hemorrhagic shock model, limited resuscitation in the prehospital phase had benefit for subsequent treatment in the hospital phase in terms of alleviated tissue damage and improved survivorship.