Effect of oxymatrine on neutrophil function based on zebrafish inflammation model and primary neutrophil inflammatory responses.

Sophora flavescens Ait. (SFA), an extensively utilized herb for the treatment of fevers, inflammatory disorders, ulcers and skin diseases related to bur, contains oxymatrine (OMT) as its principal active constituent. OMT exerts regulatory effects over inflammation, oxidative stress and apoptosis. Neutrophils, critical regulators of the inflammation response, have not been thoroughly elucidated regarding the protective properties and underlying mechanisms of OMT-mediated anti-inflammation. This study was aim to explore the protective effect of OMT on neutrophils under inflammatory conditions and delve into its potential mechanism. Leveraging the advantages of zebrafish, an animal model with a real-time dynamic observation system, we established an in vivo caudal fin wound model and a copper sulfate induced-inflammation model in zebrafish line Tg (mpx:GFP). The result revealed that OMT significantly attenuated neutrophil migration, upregulated the mRNA expression levels of JNK, casp3, mapk14a, mapkapk2a and map2k1 damaged by zebrafish caudal fin wound model, and downregulated mRNA expression levels of JNK, casp3, mapk14a, mapkapk2a and map2k1 in the copper sulfate injury model. In vitro experiments demonstrated that OMT modulated the chemotaxis response of primary neutrophils from mice, enhanced phagocytosis, reduced oxidative stress and alleviated inflammation level. We hypothesize that the OMT may exert its anti-inflammatory effects by regulating primary neutrophils through the MAPK signaling pathway.

Comparison of Protective Effect of Tri-circulator and Coenzyme Q10 on Myocardial Injury and the Mechanism Study by Zebrafish Model.

Tri-Circulator (TC) is a product comprising coenzyme Q10 (CoQ10), Salvia miltiorrhiza, and Panax notoginseng. Individually, each of these constituents has demonstrated protective effects on myocardial injury. The purpose of this study is to evaluate the protective efficacy of TC on heart function and compare the differential effects between CoQ10 and TC. Two myocardial injury models of zebrafish, the hypoxia-reoxygenation model (H/R) and the isoproterenol (ISO, a ß-receptor agonist) model, were used in this experiment. The zebrafish subjects were divided into 4 groups: control, H/R, TC, and CoQ10. Heart rate, stroke volume (SV), cardiac output (CO), ejection fraction (EF), fractional area change (FAC), and pericardial height were monitored to assess changes in heart function. The gene expression of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) was studied as markers of injury/stress. TC significantly suppresses elevated heart rate induced by H/R and prevents the decrease of heart rate induced by ISO. It alleviates the pericardial infusion induced by ISO, whereas CoQ10 does not possess a similar effect. Both TC and CoQ10 significantly inhibit the decline in SV, CO, EF, and FAC induced by H/R and ISO, and suppress the expression of ANP and BNP in cardiomyocytes induced by ISO. It is noteworthy that TC demonstrates a more pronounced effect on EF, FAC, ANP, and BNP gene expression compared to CoQ10. Both TC and CoQ10 have a protective effect on myocardial injury of zebrafish. However, TC exhibits a greater efficacy compared to CoQ10 alone in mitigating myocardial injury.

Identification of histone deacetylase inhibitors as neutrophil recruitment modulators in zebrafish using a chemical library screen.

Tissue injury-induced neutrophil recruitment is a prerequisite for the initiation and amplification of inflammatory responses. Although multiple proteases and enzymes involved in post-translational modification (PTM) of proteins regulate leukocyte recruitment, an unbiased functional screen of enzymes regulating inflammatory leukocyte recruitment has yet to be undertaken. Here, using a zebrafish tail fin amputation (TFA) model to screen a chemical library consisting of 295 compounds that target proteases and PTM enzymes, we identified multiple histone deacetylase (HDAC) inhibitors that modulate inflammatory neutrophil recruitment. AR-42, a pan-HDAC inhibitor, was shown to inhibit neutrophil recruitment in three different zebrafish sterile tissue injury models: a TFA model, a copper-induced neuromast damage and mechanical otic vesicle injury (MOVI) model, and a sterile murine peritonitis model. RNA sequencing analysis of AR-42-treated fish embryos revealed downregulation of neutrophil-associated cytokines/chemokines, and exogenous supplementation with recombinant human IL-1ß and CXCL8 partially restored the defective neutrophil recruitment in AR-42-treated MOVI model fish embryos. We thus demonstrate that AR-42 non-cell-autonomously modulates neutrophil recruitment by suppressing transcriptional expression of cytokines/chemokines, thereby identifying AR-42 as a promising anti-inflammatory drug for treating sterile tissue injury-associated diseases.

Salviae miltiorrhizae Liguspyragine Hydrochloride and Glucose Injection Protects against Myocardial Ischemia-Reperfusion Injury and Heart Failure.

Purpose: Myocardial ischemia-reperfusion (MIR) injury is a common stimulus for cardiac diseases like cardiac arrhythmias and heart failure and may cause high mortality rates. Salviae miltiorrhizae liguspyragine hydrochloride and glucose injection (SGI) has been widely used to treat myocardial and cerebral infarctions in China even though its pharmacological mechanisms are not completely clear. Methods: The protective effect and mechanism of SGI on MIR injury and heart failure were investigated through the H9c2 cell model induced by hypoxia/reoxygenation (H/R) and rapamycin, zebrafish model induced by H/R and isoprenaline, and rat MIR model. Results: SGI significantly reduced the infarct size and alleviated the impairment of cardiac functions in the MIR rat model and H/R zebrafish model and promoted cell viability of cardiomyocyte-like H9c2 cells under H/R condition. Consistently, SGI significantly downregulated the serum level of biomarkers for cardiac damage and attenuated the oxidative damage in the MIR and H/R models. We also found that SGI could downregulate the increased autophagy level in those MIR and H/R models since autophagy can contribute to the injurious effects of ischemia-reperfusion in the heart, suggesting that SGI may alleviate MIR injury via regulating the autophagy pathway. In addition, we demonstrated that SGI also played a protective role in the isoproterenol-induced zebrafish heart failure model, and SGI significantly downregulated the increased autophagy and SP1/GATA4 pathways. Conclusion: SGI may exert anti-MIR and heart failure by inhibiting activated autophagy and the SP1/GATA4 pathway.

Anti-obesity effects of galacto-oligosaccharides in obese rats.

Galacto-oligosaccharides (GOS) are commonly used as prebiotic with a variety of known metabolic benefits; however, whether GOS plays a protective role in obesity remains unknown. Here, we demonstrate that GOS prevented obesity in a rat model of obesity induced by a high-fat diet. Our results showed that GOS effectively slowed weight gain in diet-induced obese rats without affecting energy intake. GOS significantly suppressed the hypertrophy and hyperplasia of white adipose tissue and markedly reduced the ratio of the fat/body. Consistently, GOS significantly improved serum total cholesterol, triglycerides, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels, indicating the weight loss activity of GOS. Interestingly, GOS also significantly increased the expression levels of browning proteins, including uncoupling protein 1, peroxisome proliferator-activated receptor-γ, peroxisome proliferator-activated receptor-γ coactivator 1α, and PR domain 16, in both white and brown adipose tissue. Furthermore, we found that GOS markedly increased the expression levels of liver X receptor α, peroxisome proliferation-activated receptor-α, low-density lipoprotein receptor, and cholesterol 7α-hydroxylase proteins in the liver of obese rats. Taken together, we concluded that GOS inhibits obesity by accelerating the browning of white fat cells and the thermogenesis of brown fat cells and that GOS improves host lipid homeostasis by promoting cholesterol catabolism.

Diagnostic Value of Circular RNA hsa_circ_0002874 Expression in Peripheral Blood of Patients with Gastric Cancer.

The purpose of this study was to determine whether circular RNA hsa_circ_0002874 could serve as a novel biomarker for the diagnosis of gastric cancer (GC). The expression level of hsa_circ_0002874 mean (interquartile range [IQR]) in the plasma of patients with GC, patients with benign gastric lesions, and healthy individuals was 3.482 (IQR, 1.524-9.048), 1.261 (IQR, 0.817-2.000), and 1.00 (IQR, 0.726-1.382), respectively, whereas there was no significant difference between the latter 2 groups. The plasma expression level of hsa_circ_0002874 was significantly correlated with tumor stage (U = 234.0; P < .001) and lymph node metastasis (U = 240.0; P < .001). The receiver operating characteristic (ROC) curve showed that the sensitivity of the combined determination of hsa_circ_0002874 and the serum markers CEA and CA19-9 was 95.8% in patients with GC compared with that of the healthy group and 93.0% compared with that of patients with benign gastric tumor lesions. The specificity of hsa_circ_0002874 in differentiating GC from benign lesions was 98.3%. The results showed that plasma hsa_circ_0002874 may prove to be a useful biomarker for auxiliary diagnosis, the grading of malignant neoplasms, and the prognostic prediction of GC.

The effects of COST on the differentiation of 3T3-L1 preadipocytes and the mechanism of action.

The objectives of this study were to explore the effect of COST (one thousand Da molecular weight chitosan oligosaccharide) on the differentiation of 3T3-L1 preadipocytes and to determine the mechanism of action. 3T3-L1 preadipocytes were used as the target cells, and the induction of the methods for the differentiation of 3T3-L1 preadipocytes was based on classic cocktails. The MTT assay was used to filtrate the concentration of COST. On the 6th day of induced-differentiation, the differentiation of 3T3-L1 cells was detected by Oil Red O staining. The expression of PPARγ and C/EBPα mRNA was determined using real-time fluorescence quantitative PCR (Q-PCR). COST inhibited 3T3-L1 preadipocyte differentiation in a dose-dependent manner and decreased lipid accumulation. At the molecular level, the expression of the transcription factors, PPARγ and C/EBPα, was reduced by COST during adipogenesis. These results indicate that COST effectively inhibited the differentiation of 3T3-L1 preadipocytes. The mechanism is related to the down-regulation expression of PPARγ and C/EBPα.

Closed-circuit anesthesia prolongs the neuromuscular blockade of rocuronium.

BACKGROUND: Volatile anesthetics are known to potentiate the neuromuscular blocking effect of nondepolarizing muscle relaxants. The influences of anesthetic techniques, closed-circuit anesthesia (CCA) and high flow semi-closed anesthesia (SCA), on the neuromuscular blockade of rocuronium has not yet been studied in detail. This study was purposed to compare the effects of isoflurane conveyed in minimal flow (CCA) and in high flow (SCA) on the neuromuscular blockade of rocuronium. METHODS: Fifty females scheduled for elective laparoscopic gynecological surgery were enrolled for study and randomly assigned to receive either CCA (n = 25) or SCA (n = 25). Anesthesia was induced with fentanyl 2 micrograms/kg, thiopental 5 mg/kg and rocuronium 0.6 mg/kg. Two percent isoflurane in high O2 flow (3 l/min) was given for 10 min to all patients initially to wash isoflurane in the functional residual capacity of both lungs and the breathing circuit. After the wash in, for CCA group, the O2 flow was reduced to 300 ml/min with isoflurane vaporizer setting adjusted to 3-5% for anesthesia maintenance, while for SCA group, anesthesia was maintained with 1.5-2% isoflurane in 3 l/min O2 flow throughout the surgery. Electromyogram was used to determine neuromuscular blockade. Rocuronium (0.15 mg/kg) was given to maintain muscle relaxation when T1 reached 25% of control. We maintained the anesthetic depth until the recordings of T1 twitch response which reached 75% was completed. Onset time, duration, recovery index and intubating conditions were recorded. The hemodynamic parameters and the inhaled/exhaled concentrations were also measured every 15 min after skin incision in both groups. RESULTS: The onset time and intubating conditions were similar in both groups. In comparison with SCA group, longer clinical durations (54.1 +/- 14.4 vs. 45.4 +/- 9.2 min, P < 0.05), longer durations of maintained dose (41.1 +/- 11.1 vs. 30.2 +/- 8.6 min, P < 0.01) and longer recovery index (34.2 +/- 10.7 vs. 20.9 +/- 5.4 min, P < 0.0001) were observed in CCA group. CONCLUSIONS: We conclude that CCA may further prolong the neuromuscular blocking effect of rocuronium than SCA.

Intracranial subdural hematoma after unintended durotomy during spine surgery.

PURPOSE: To report a case of intracranial subdural hematoma occurring after a spinal dural tear that was made unintentionally during the course of a posterior laminectomy and spinal fusion at the L(5)-S(1) level. The possible physiopathological mechanisms are discussed. CLINICAL FEATURES: On the fourth postoperative day, a 59-yr-old woman displayed persistent headache following unintended durotomy during spine implant revision. Perioperative blood loss was 2840 mL and intravascular replacement was about 3000 mL. She was hydrated with iv fluids and treated with non-steroidal anti-inflammatory drugs. The symptoms improved but persisted. With the aggravation of the headache complicated with unconsciousness and the appearance of focal neurological signs on the eighth day, a computed tomography was obtained and revealed a right subdural hematoma. Following surgical drainage, the patient made an uneventful recovery. CONCLUSION: This case reminds us that subdural hematoma formation can complicate durotomy during spine surgery. Neurological deterioration in the postoperative period should prompt clinicians to rule out the diagnosis and intervene rapidly as appropriate.