Naringin alleviates pneumonia caused by Klebsiella pneumoniae infection by suppressing NLRP3 inflammasome.

Klebsiella pneumoniae (Kpn) is an important pathogen of hospital-acquired pneumonia, which can lead to sepsis and death in severe cases. In this study, we simulated pneumonia induced by Kpn infection in mice to investigate the therapeutic effect of naringin (NAR) on bacterial-induced lung inflammation. Mice infected with Kpn exhibited increases in white blood cells (WBC) and neutrophils in the peripheral blood and pathological severe injury of the lungs. This injury was manifested by increased expression of the inflammatory cytokines interleukin (IL)- 18, IL-1ß, tumor necrosis factor-α (TNF-α) and IL-6, and elevated the expression of NLRP3 protein. NAR treatment could decrease the protein expression of NLRP3, alleviate lung inflammation, and reduce lung injury in mice caused by Kpn. Meanwhile, molecular docking results suggest NAR could bind to NLRP3 and Surface Plasmon Resonance (SPR) analyses also confirm this result. In vitro trials, we found that pretreated with NAR not only inhibited nuclear translocation of nuclear factor (NF)-κB protein P65 but also attenuated the protein interaction of NLRP3, caspase-1 and ASC and inhibited the assembly of NLRP3 inflammasome in mice AMs. Additionally, NAR could reduce intracellular potassium (K+) efflux, inhibiting NLRP3 inflammasome activation. These results indicated that NAR could protect against Kpn-induced pneumonia by inhibiting the overactivation of the NLRP3 inflammasome signaling pathway. The results of this study confirm the efficacy of NAR in treating bacterial pneumonia, refine the mechanism of action of NAR, and provide a theoretical basis for the research and development of NAR as an anti-inflammatory adjuvant.

Significance of serum connective tissue growth factor in patients with hepatocellular carcinoma and relationship with angiogenesis.

BACKGROUND: Connective tissue growth factor (CTGF) expression changes variously in different human malignancies. The role of CTGF in hepatocellular carcinoma (HCC) is still not clear. The purpose of this study was to investigate the role of serum CTGF in patients with HCC and its correlation with HCC angiogenesis. METHODS: CTGF, vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) were measured by ELISA in preoperative sera of 88 patients with HCC with tumor resection and 39 healthy subjects. The relationship between CTGF and HCC clinicopathological parameters was observed. Prognostic significance of CTGF for survival of patients with HCC was assessed by Kaplan-Meier analysis. RESULTS: Preoperative serum CTGF level was significantly higher in patients with HCC than in healthy subjects (median, 63.5 vs. 11.4 ng/ml; P < 0.001). Serum CTGF correlated significantly with a series of clinicopathological parameters (big tumor size, advanced pathological tumor-node-metastasis stage, absence of tumor capsule, portal vein invasion). Serum CTGF showed a significant correlation with disease-free survival and overall survival of patients with HCC. Patients with high serum CTGF (>63.5 ng/ml) had a poorer disease-free survival time than the others (CTGF < or = 63.5 ng/ml; median disease-free survival time, 9.6 vs. 19.3 months). Patients with high serum CTGF had poorer overall survival time (median, 13.1 months) than the others (median, 21.7 months). In multivariate Cox analysis, CTGF was identified as an independent and significant prognostic factor of survival of HCC patients. CONCLUSIONS: Serum CTGF plays an important role in the progression of HCC. Serum CTGF may be a potential indicator of angiogenesis of HCC.

Cell death-inducing DFF45-like effector may take part in neuronal apoptosis of the lumbar spinal cord after sciatic nerve injury caused by a firearm.

OBJECTIVE: To investigate neuronal apoptosis and gene regulation after peripheral nerve injury caused by a firearm. METHODS: A rabbit model of sciatic nerve injury caused by a firearm was used. Neuronal apoptosis was determined by using terminal transferase-mediated dUTP nick end-labeling and flow cytometry. Differentially expressed genes were screened with mouse_8192S GeneChips. Cell death-inducing DFF45-like effector (CIDE-B) gene expression in spinal cord after sciatic nerve injury was detected with reverse transcription-polymerase chain reaction. RESULTS: The size of the injury caused by the firearm was widely extended. Neuronal apoptosis was found in days 1, 3, and 7 groups. The expression of apoptosis-related gene CIDE-B was high according to analysis of the GeneChips and reverse transcription-polymerase chain reaction detection. The trend of CIDE-B expression was consistent with neuronal apoptosis after nerve injury. CONCLUSIONS: Neuronal apoptosis is an important pathological change after peripheral nerve injury caused by firearms, and CIDE-B may be an important regulator.

EVALUATION OF THE ACUTE AND SUBCHRONIC TOXICITY OF Aster tataricus L. F.

BACKGROUND: Aster tataricus L. f. is used as a traditional Chinese drug to relieve cough and asthma symptoms and to eliminate phlegm. However, Aster tataricus L. f. possesses toxicity, and little systematic research has been conducted on its toxic effects in the laboratory. METHODS AND MATERIALS: The acute group was administered 75% alcohol extract of Aster tataricus L. f. in a single dose. A subchronic toxicity study was performed via daily oral administration of Aster tataricus L. f. at a dose of 0.34 g/kg body weight in SD rats. The rats were divided into six groups: a petroleum ether extract (PEA) group, an ethyl acetate extract (EEA) group, an n-butyl alcohol extract (NEA) group, a remaining lower aqueous phases (REA) group, a 75% alcohol extract (AEA) group and a control group. Quantitative measurements of cytokines were obtained by fluorescence with a laser scanner using a Cy3 equivalent dye. RESULTS: The LD50 of the 75% alcohol extract of Aster tataricus L. f. was 15.74 g/kg bw. In the subchronic toxicity study, no significant differences were observed among groups in relative organ weights, urine traits, liver antioxidase levels, or cytokine levels. However, significant sporadic differences were observed in body weight gains, haematology indices, biochemistry values, and histopathology features in PEA, EEA group. In addition, sporadic changes in other groups in measures such as WBC, MCHC, CK, ALP, AST, ALT, LDH, T-BIL, LDL-C, HDL-C, and TC were observed. CONCLUSION: The toxicity study showed that Aster tataricus L. f. can produce toxic effects, mainly on the liver; much less on the heart. The LD50 was 15.74 g/kg BW in mice, and the subchronic toxicity study, used a dosage of 0.34 g/kg/d.BW, showed that the toxic components of Aster tataricus L. f. were mainly concentrated in the petroleum ether fraction, followed by the ethyl acetate fraction, the n-butyl alcohol fraction, the lower aqueous phase and the 75% ethanol extracts. Abbreviations: PEA, petroleum ether extract of Aster tataricus L. f.; EEA, ethyl acetate extract of Aster tataricus L. f.; NEA: n-butyl alcohol extract of Aster tataricus L. f.; REA: lower aqueous phases of Aster tataricus L. f.; AEA, 75% alcohol extract of Aster tataricus L. f.; WBC, white blood cell; RBC, red blood cell, PLT, platelet; HCT, haematocrit; MCV, mean corpuscular volume; HGB, haemoglobin; MCH, mean corpuscular haemoglobin; MCHC, mean corpuscular haemoglobin concentration; CREA, creatinine; LDH, lactate dehydrogenase; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol; T-BIL, total bilirubin; ALT, alanine aminotransferase; ALP, alkaline phosphatase; AST, aspartate aminotransferase; TP, total protein; ALB, albumin; Glu, glucose; TC, total cholesterol; TG, triglycerides; CK, creatine kinase; GSH, Glutathione; MDA, malondialdehyde; T-SOD, total superoxide dismutase; TNF, tumour necrosis factor; IFN, interferon; MCP, monocyte chemotactic protein C.

Identification of differentially expressed genes from spinal cord after firearm injury to rabbit sciatic nerves.

OBJECTIVE: Peripheral nerve regeneration depends on gene regulation by central neurons. To search for more effective treatment methods to improve the regeneration of wounded peripheral nerves, gene expression profile of spinal cord after firearm injury to rabbit sciatic nerves are studied with DNA micro-array technique. METHODS: A total of 54 rabbits were randomly divided into 4 groups: Groups d1, d3, d7 and normal control group. Lumbar spinal cords were sampled. RNA and mRNA were extracted, labeled by Cy3 and Cy5, and analyzed by mouse_8192S gene chips. RESULTS: A total of 1367, 923, and 61 genes with differential expression were found on day 1, day 3, and day 7 after trauma respectively. Five expressed sequence tag (EST) sequences demonstrated differential expression during 7 days after trauma. CONCLUSIONS: There is complex gene profile with differential expression after firearm nerve injury, among which AW701496, U84291, W13926, X04017 and AW822394 EST sequences may be important regulation factors that involved in regeneration of peripheral nerve injury.

Effect of artesunate on endotoxin-induced uveitis in rats.

PURPOSE: The purpose of this study was to explore the effect of artesunate on endotoxin-induced uveitis (EIU) in rats. METHODS: EIU was induced in male Long-Evans rats by footpad injection of lipopolysaccharide (LPS 200 µg in 0.1 mL PBS; LPS-only group [LG]). PBS (0.1 mL), prednisolone (10 mg/kg in 0.1 mL PBS; prednisolone group [PG]), and artesunate of three concentrations (1, 10, 100 mg/kg; artesunate groups A, B, C [AGA, AGB, AGC]; all in 0.1 mL of PBS) were injected intravenously, respectively, 1 hour after LPS injection. Twenty-four hours after LPS injection, infiltrating cells and protein concentration in the aqueous humor, as well as tumor necrosis factor (TNF)-α, nitric oxide (NO), prostaglandin E2 (PGE2), and monocyte chemoattractant protein (MCP)-1, were measured. RESULTS: Rats in AGA did not have significant difference in infiltrating cell number (61.6 × 10(5) cells/mL; range, 20.0-147.8 cells/mL; P = 0.287), protein concentration (48.0 mg/mL; range, 18.0-101.8 mg/mL; P = 0.349), NO concentration (86.9 µM/L; range, 30.5-192.5 µM/L; P = 0.363), but had significant difference in TNF-α (1144.3 pg/mL; range, 460.0-1840.0 pg/mL; P = 0.038), PGE2 (12.8 ng/mL; range, 4.8-27.2 ng/mL; P = 0.005), and MCP-1 (6136 pg/mL; range, 20.0-147.8 pg/mL; P = 0.009) with those of LG. Artesunate at 10 mg/kg and 100 mg/kg significantly suppressed the infiltrating cells, protein concentration, TNF-α, NO, PGE2, and MCP-1 in the aqueous humor induced by LPS (P = 0.011-0.000). CONCLUSIONS: Dose-dependent decreases of infiltrating cells, protein concentration, TNF-α, PGE2, NO, and MCP-1 in the aqueous humor by artesunate treatment after LPS injection indicate that artesunate can suppress the inflammation of EIU by inhibiting the production of inflammatory mediators.

Relationship between serum heparanase and microscopic venous invasion in patients with hepatocellular carcinoma.

The serum heparanase level of 92 patients with hepatocellular carcinoma (HCC) measured by enzyme-linked immunosorbent assay (median, 91.4 U/mL) was higher than that of 19 healthy control subjects. Serum heparanase levels were higher in patients with HCC characterized by large tumors (>5 cm), advanced pTNM stage (III and IV), tumor capsule absence, and portal vein invasion. Positive correlations between serum heparanase and tumor heparanase expression were observed in 92 patients with HCC, 53 among them treated with tumor resection. In these 53 patients, the rate of microscopic venous invasion was significantly higher in 18 cases with high serum heparanase levels (>91.4 U/mL) than in the other 35 cases with low heparanase levels (14/18 vs 10/35; P = .001). Serum heparanase and vascular endothelial growth factor were identified as independent predictive factors for HCC microscopic venous invasion. The postoperative recurrence-free time (median, 8.7 months; range, 4.1-22.9 months), recurrence rate (72% [13/18]), 1.5-year disease-free survival rate (29.7%) were significantly worse in these 18 patients.