AIM and METHODES: To evaluate the possible signal transduction mechanism of nontargeted mutagenesis in vero cells induced by DNA damaging agent N-methyl-N-nitro-N-nitrosoguanidine(MNNG),the activation of c-Jun NH 2-terminal kinase/stress activated protein kinase(SAPK/JNK) pathway in vero cells induced by MNNG was studied. Western Blot analysis and Solid-phase kinase assay were used to measure the phosphorylation of JNK1 and kinase activity of JNKs, respectively. RESULTS: After 0.2 ?mol/L, 2.5 h MNNG or 1 mg/L, 1 h cycloheximide (CHM) treatment, the proportion of phosphorylated JNK1 in cell extract increased significantly, simultaneously the kinase activity of JNKs increased dramatically(6.7 and 3.0 folds respectively), as measured by the phosphorylation of c-Jun, a substrate of JNKs. CONCLUSION: Both 0.2 ?mol/L 2.5 h MNNG and 1 mg/L 1 h CHM treatment can induce the activation of JNK/SAPK pathway, one of the stress signal transduction pathways, in vero cells.

Background:Faecalibacterium prausnitzii( Fp) is one of the most abundant bacterium in human intestinal microbiota,and is closely correlated with the process of colitis-associated colorectal cancer(CAC). Aims:To observe the effect of Fp on CAC,and investigate the possible mechanism. Methods:The model of CAC was induced by azoxymethane (AOM)and dextran sodium sulfate( DSS). Fifty-two C57BL/ 6J mice were randomly divided into 4 groups:group A (AOM + DSS),group B(AOM + DSS + Fp),group C(AOM + DSS + Fp supernatant)and group D(control group). All the mice were sacrificed on day 92. DAI was assessed,serum levels of TNF-α and IL-10 were determined by ELISA. HE staining was used to examine the grade of tumor. Expressions of VEGF,COX-2,NF-κB in tumor tissue were measured by immunohistochemistry. Results:The tumorigenesis rates of group A,B,C were 100% ,100% and 77. 8% ,respectively;mainly were high-grade intraepithelial neoplasia. The tumor load in group A was significantly higher than that in group B (P < 0. 01),and the spleen index in group B was significantly higher than that in group C(P < 0. 01). Serum level of TNF-α was significantly lower(P < 0. 05)and IL-10 was significantly higher(P < 0. 05)in group A than that in group B. No significant differences in expressions of VEGF,COX-2,NF-κB were found among group A,B and C. Conclusions:Fp had no obvious effect on the occurrence rate of CAC,and Fp supernatant could decrease the incidence of CAC in mice. Fp and its supernatant could reduce the tumor load via regulating the expressions of TNF-α,IL-10.

ObjectiveTo investigate the effect of Naoxueshu oral liquid on cerebral hemorrhage caused by hypertension. Methods140 patients with cerebral hemorrhage were randomly dividied into two groups with a 1∶1 ratio,treatment group and control group. Patients in treatment group received Naoxueshu oral liquid 30 ml daily,while that in control received Naoxuekang oral liquid 30 ml daily. The changes of score on Chinese medical integration,National Institutes of Health (NIH) Stroke scale and the volume of intracranial hemorrhage were evaluated before and a month after treatment.ResultsThe decrease of NIH score of patients in treatment group was (12.73±3.94),but(4.72±3.01) with the control's (t=13.5327,P<0.001). The decrease of the volume of intracranial hemorrhage of patients in treatment group was (13.28±4.17) ml,but (8.37±7.24) ml with the control's(t=4.9228,P<0.001). ConclusionComparing with Naoxuekang,Naoxueshu can accelerate absorption of hematome and ameliorate the recovery of neurological disability more efficiently.

Objective To evaluate the effect for the treatment of severe asthma. Methods The data of 47 patients with severe asthma who were admitted to emergency department were retrospectively anayzed. Results Of total 47 patients ,45 were rescued, with the survival rate of 95.7%. Arterial blood gas was improved after treatment (P < 0.01). Conclusion Appropriate commencement, mode, strategy, and early weaning of mechanical ventilation, combined with the administration of bronchodilators and eorticosteroids are the important way to rescue patients with severe asthma.

Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; therapeutic use ; Hypolipidemic Agents ; therapeutic use ; Pharmacogenetics

Objective To evaluate the role of nuclear factor erythroid 2?related factor 2 ( Nrf2)∕antioxidant response element( ARE) signaling pathway in inhibition of lipopolysaccharide ( LPS)?induced inflammatory factor release from macrophages by hydrogen. Methods RAW264. 7 macrophages of mice were cultured in 6?well plates (2×106 cells∕well) and were divided into 4 groups (n=24 each) using a random number table: control group ( group C); LPS group; hydrogen?rich saline+LPS group ( group LPS+H2); Nrf2 small interference RNA (siRNA)+LPS+hydrogen?rich saline group (siRNA+LPS+H2 group) . LPS 1 μg∕ml was added in group LPS. In group LPS+H2 , LPS 1μg∕ml was added, and the cul?ture medium was then replaced with the culture medium containing 0. 6 mmol∕L hydrogen?rich saline. In group siRNA+LPS+H2 , after Nrf2?siRN was successfully transfected into the cells, the cells were continu?ously incubated for 24 h, and the culture medium was then replaced with the culture medium containing 0.6 mmol∕L hydrogen?rich saline after LPS 1 μg∕ml was added. At 24 h of incubation, the supernatant was sep?arated for determination of the lactic dehydrogenase (LDH) activity (using colorimetric method) and for detection of the concentrations of tumor necrosis factor?alpha ( TNF?α) , interleukin?1 beta ( IL?1β) , high mobility group box?1 (HMGB1) and IL?6 (by ELISA). The cells were collected for measurement of the proliferation of cells ( by methyl thiazolyl tetrazolium assay) and for determination of the expression of Nrf2 and heme oxygenase?1 ( HO?1) in cells ( by Western blot) . Results Compared with group C, the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly in?creased, the proliferation of cells was significantly decreased, and the expression of HO?1 in cells was sig?nificantly up?regulated in LPS and siRNA+LPS+H2 groups, and the expression of Nrf2 in cells was signifi?cantly up?regulated in LPS and LPS+H2 groups (P<0.05). Compared with group LPS, the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly decreased, the proliferation of cells was significantly increased, and the expression of Nrf2 and HO?1 in cells was sig?nificantly up?regulated in group LPS+H2 , and the expression of Nrf2 and HO?1 in cells was significantly down?regulated in group siRNA+LPS+H2 ( P<0.05) . Compared with group LPS+H2 , the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly increased, the proliferation of cells was significantly decreased, and the expression of Nrf2 and HO?1 in cells was signifi?cantly down?regulated in group LPS+H2+siRNA ( P<0.05) . Conclusion The mechanism by which hydro?gen inhibits LPS?induced inflammatory factor release from macrophages is related to the activation of Nrf2∕ARE signaling pathway in mice.

Objective To establish the HPLC fingerprint of Herba Selaginellae moellendorfii for identification and comparison with other species of the same genus and to determine the content of amentoflavone.Method Separation was performed on ZORBAX SB-C18 chromatographic column,acetonitrile(A)-0.5 %solution of acetic acid in water(B)as mobile phase with gradient elution and the flow rate was 1.0 mL?min-1.The detection wavelength was at 270 nm.The content determination of amentoflavone carried out synchronously.Results The characteristic eighteen peaks in the chromatogram consisted of the common pattern of Herba Selaginellae moellendorfii.The fingerprint coupling with similarity and Principal Component Analysis differentiated and classified the various species of selaginella.Nine species could be divided into 3 classes.The content of amentoflavone in Herba Selaginellae moellendorfii was 0.8～1.0 %.Conclusions The weighted similarity coefficient was calculated from 13 batches of Herba Selaginellae moellendorfii samples as high as more than 0.98.Among 9 species of Selaginella,5 species(S.biformis,S.involvens,S.doederileinii,S.trachyphylla,S.tamariscina)were sorted out by means of Principal Component Analysis in the same class with S.moellendorfii.Which indicated the possibility of bio-equivalence among the herbs of these six species,while S.picta,S.uncinata and S.delicatula are considered as adulterants of S.moellendorfii as their dissimilar fingerprints.

Objective To prepare polyclonal antibodies of anti Nogo-66, the extracellular region of one central nervous system neurite regeneration inhibitor Nogo, which could be used to further identification and functional study of Nogo molecule.Methods Preparing rabbit anti rat Nogo-66 polyclonal antibodies with a purified Nogo-66 fusion protein expressed in E.coli system. Studying its specificity by Western-blot and immuno-histochemical techniques and identifying its biological activity in PC12 cells.Results The high titer (1∶[KG-*2]10 000) anti rat Nogo-66 polyclonal antibodies were obtained.This antibody could specifically recognize the Nogo protein expressed in E.coli system.Immuno-histochemical staining indicated that the Nogo was widely expressed in rat spinal cord neurons and oligodendrocytes.It could effectively block the neurite extensioninhibition of Nogo protein in PC12.Conclusion Successful preparation of anti rat Nogo polyclonal antibodies provides a useful tool in identification or further functional study of Nogo molecule.

Objective To investigate the expression of vascular endothelial cell growth factor(VEGF) and its upregulation of matrix metalloproteinase-2(MMP-2)activation in synovial fluid of patients with rheumatoid arthritis(RA)as well as its role in the pathogeneses of RA.Mathods Expression of VEGF and receptor KDR in mononuclear cell(MNC)of synovial fluid of RA patients and controls were determined by Western blot;VEGF levels of supernantants from MNC was determined by ELISA;supernantants from KDR~+ MNC of synovial fluid of RA patients collected after incubation in serum-free medium with or without VEGF, their activity of MMP-2 was measured by gelatinolytic zymography;Boyden chamber-matrigel in vitro invasion assay was used to detect the invasive capacity in vitro in KDR~+ MNC of synovial fluid of RA patients incubat- ed with or without VEGF.Results The expressions of VEGF/KDR in MNC of synovial fluid of RA patients were significantly higher than those of controls;the MMP-2 activity and invasive ability of co-cuhured KDR~+ MNC with VEGF was higher than those of without VEGF.Conclusion VEGF upregulates MMP-2 activation and promotes invasion of MNC of synovial fluid of RA patients by interacting with receptor KDR,indicating that VEGF plays an important role in RA pathology.

Objective To evaluate the predicting value of serum procaleitonin (PCT) in treatment of acute exacerbations of chronic obstructive pulmonary disease (AECOPD) in elderly patients. Methods A total of 267 elderly patients requiring hospitalization for AECOPD were randomly assigned into 2 groups: standard therapy group (standard group, n= 135) and PCT-guided group(PCT group, n= 132). Standard group received antibiotics according to the guideline of attending physicians and PCT group were treated with antibiotics according to serum PCT levels.Length of hospitalization, clinical efficacy, costs of hospitalization and antibiotics, rate of antibiotics use, hospital mortality, rate of exacerbation and rehospitalization, frequency of exacerbation within 1 year were observed. Results Length of hospitalization, clinical efficacy, hospital mortality, rate of exacerbation and rehospitalization, frequency of exacerbation within 1 year were similar in 2 groups(all P＞0.05);costs of hospitalization and antibiotics, rate of antibiotics use of PCT group[10 882 (3808-16 651)yuan, 6934 (2390-10 660)yuan, 76.5%] were lower than those of standard group[13 637(4650-19 730)yuan, 8589(3144-12 117)yuan, 87.4%] (all P＜0.05). Conclusions PCT guidance offers an advantage over standard therapy in reducing antibiotic use and in lowering the costs of hospitalization in treatment of AECOPD in elderly patients.