Spatiotemporal expression of inducible nitric oxide synthase and cyclooxygenase 2 in the spinal cord during early stage sciatic nerve crush injury

BACKGROUND： Previous studies have shown that inducible nitric oxide synthase （iNOS） and cyclooxygenase 2 （COX-2） participate in inflammatory immune responses and neuropathic pain following peripheral nerve injury. However, few reports have addressed time-dependent expression of iNOS and COX-2 following peripheral nerve injury. OBJECTIVE： To investigate spatiotemporal expression of iNOS and COX-2 during early stage sciatic nerve crush injury.DESIGN, TIME AND SETTING： The randomized, controlled, animal experiment was performed at the Laboratory of Applied Anatomy, Department of Human Anatomy and Neurobiology, Central South University, China from September 2006 to September 2007.MATERIALS： Mouse anti-rat iNOS monoclonal antibody and goat anti-rat COX-2 monoclonal antibody （Transduction Laboratory, USA）, as well as biotinylated rabbit anti-mouse lgG and biotinylated rabbit anti-goat IgG （Santa Cruz Biotechnology, USA） were used in the present study.METHODS： A total of 48 healthy, adult, Sprague Dawley rats were randomly assigned to three groups. In the model group （n = 32）, crush injury to the right sciatic nerve was established using an artery clamp. The model group was further assigned to four subgroups according to survival time （6,12, 24, and 72 hours）, respectively （n = 8）. Sham surgery （n = 8） and normal control （n = 8） groups were also established.MAIN OUTCOME MEASURES： iNOS and COX-2 expression was detected in the L4-6 spinal cord with immunohistochemistry. Gray values of iNOS- and COX-2-postive cells in the anterior horn and posterior horn of spinal cord, as well as quantification of iNOS- and COX-2-positive cells in the anterior horn of spinal cord, were measured.RESULTS： iNOS and COX-2 expression gradually increased in the anterior horn and posterior horn of the spinal cord on the damaged side over time from 6 hours following sciatic nerve injury （P〈0.05） and peaked at 72 hours. Simultaneously, the number of iNOS- and COX-2-positive cells similarly increased in the anterior horn of spinal cord on the damaged side （P〈 0.05）.CONCLUSION： iNOS and COX-2 expression increased in the spinal cord during early stage sciatic nerve crush, which suggested that iNOS and COX-2 participate in occurrence and development of inflammatory immune responses following peripheral nerve injury.

Expression and Significance of Cyclooxygenase 2 Gene in Lung Cancer

To study the expression of cyclooxygenase 2 (COX-2) gene and its relationship with clinicopathological characteristics of lung cancer, expression of the COX-2 mRNA was evaluated by reverse transcription polymerase chain reaction (RT-PCR) in cancerous tissues and paired adjacent non-cancerous tissues from 56 patients and benign lesions from 12 patients. Our results showed that expression of COX-2 gene was detected in a significantly greater proportion of cancerous tissues (60.7 %) than adjacent noncancerous tissues (10.7 %, P<0.01) and benign lesions (3/12, P<0.05). Expression of COX-2 gene was higher in adenocarcinoma than in squamous carcinoma (P<0.01). There was no significant relationship between COX-2 gene expression and patients' age, sex, histological type of tumors, differentiation degree and TNM stages (P>0.05). The up-regulation of COX-2 gene in lung cancer tissues especially in adenocarcinoma suggested that COX-2 may play a role in the lung carcinogenesis and COX-2 gene may serve as a potential therapeutic target in lung cancer.

Aspirin inhibits the proliferation of tobacco-related esophageal squamous carcinomas cell lines through cyclooxygenase 2 pathway

Background Cigarette smoking has been verified as the risk factor of esophageal squamous cell carcinoma （ESCC）. Overexpression of cyclooxygenase 2 （COX-2） is shown in ESCC. The objective of this study was to investigate the effects of cigarette smoking ethanol extract （EE） on the proliferation of the human ESCC cell lines, and to explore the correlation between the proliferation rate of human ESCC cell lines and the expression pattern of COX-2. Whether aspirin can inhibit the proliferation of the ESCC cell lines pretreated with EE, and regulate the mRNA expression levels of COX-2 are also examined. Methods Two human ESCC cell lines were selected. EC109 was poorly differentiated and EC9706 was highly differentiated. EC109 and EC9706 were treated with EE and aspirin for different time course. The cell growth of ESCC was measured by MTT reduction assay and the expression of COX-2 was measured by RT-PCR and Western blot analysis. Results EE promoted the proliferation of EC109 and EC9706 in dose- and time-dependent manners. In the concentration range （10-100 pg/ml for EE） and in the time range （24-72 hours） after addition of EE, the cell proliferation was prominent in an up-scaled manner respectively. Aspirin could inhibit the proliferation of cell lines EC109 and EC9706 pretreated with EE for 5 hours, in a dose-dependent manner. In the concentration range （0.5-8.0 mmot/L for aspirin）, the cell growth inhibition was prominent in an up-scaled manner accordingly （P〈0.05）. The effect of EE on cell proliferation was correlated with the up-regulation of COX-2 gene. However, the cell growth inhibition of aspirin was correlated with the down-regulation of COX-2 gene. Conclusions EE can stimulate the proliferation of human ESCC cell lines EC109 and EC9706, most likely through up-regulating the expression of COX-2. Aspirin can inhibit the proliferation of ESCC cell lines induced by EE, which suggests it may be advantageous in the chemoprevention and therapy of human tobacco-related ESCC. And its effect is likely to be related with modulating COX-2 activity.

Effect of Propyl Gallate on Activity of Cyclooxygenase 1 and 2 in Mice's Peritoneal Macrophages*

Objective: To investigate the effect of Red Peony 801 (propyl gallate,PrG) on cyclooxygenase (COX) activity in murine peritoneal macrophages. Methods: A screening model for COX inhibitors in vitro based on murine peritoneal macrophages was used. COX-1 activity was reflected by the level of 6-ketoprostaglandin F1α(6-keto-PGF1α) in supernatants of cultured macrophages which were stimulated with calcium ionophore A23187 for a short-term, while COX-2 activity was reflected by the level of prostaglandin E2 (PGE2) in supernatants of cultured macrophages which were stimulated with lipopolysaccharide (LPS) for a long-term. Results: PrG did not affect A23187-induced, COX-1-derived 6-keto-PGF1α synthesis at the concentrations of 1×10-5, 5×10-6 mol/L (P>0.05), but enhanced 6-keto-PGF1α synthesis at the concentrations of 1×10-6, 5×10-7, 1×10-7 mol/L (P<0.01) in vitro, and showed a good dose-dependent manner. It inhibited LPS-induced, COX-2-derived PGE2 synthesis at the concentrations of 1×10-5,1×10-6 mol/L (P< 0. 05). Conclusion: Within the range of 1×10-5 to 1×10-7 mol/L, PrG activated COX-1 at lower concentrations and inhibited COX-2 at higher concentrations in murine peritoneal macrophages.

Tolerance of neurite outgrowth to Rho kinase inhibitors decreased by cyclooxygenase-2 inhibitor

In this study, PC12 Adh cells and Neuro-2a cells were treated with Rho-associated kinase inhibitors （Y27632 and Fasudil）, a cyclooxygenase-1 selective inhibitor （SC560）, and a cyclooxygenase-2 inhibitor （NS398）. We found that these cells became tolerant to Rho-associated kinase inhibitors, as neurite outgrowth induced by these inhibitors diminished following more than 3 days of exposure in either cell line. The proteins cyclooxygenase-2 and cytosolic prostaglandin E synthetase were upregulated at day 3. NS398 decreased the tolerance to neurite outgrowth induction in both cell lines, whereas SC560 had almost no effect. These findings indicate that cells become tolerant to neurite outgrowth induced by Rho-associated kinase inhibitors, this is at least partly associated with upregulation of proteins involved in the cyclooxygenase-2 pathway, and cyclooxygenases-2 inhibition prevents this tolerance.

Quantitative analysis of cyclooxygenase 2 in the posterior longitudinal ligament of cervical spondylotic myelopathy

Background Cervical spondylotic myelopathy （CSM）, in part, results from degeneration of the posterior longitudinal ligament （PLL）, which mechanically compresses the spinal cord. Much research was done on the ossification of PLL, but not concerning the non-ossifying degeneration of cervical PLL. The degeneration of cervical PLL may be related to inflammation. The aim of this study was to elucidate the pathological features of the PLL and the role of cyclooxygenase 2 （COX-2） in the degeneration of the PLL in CSM. Methods A total of 23 PLL specimens were collected during surgery from patients with CSM for the histological and immunohistochemical （type II collagen and Ki-67） study. For the control group 14 cervical PLL autopsy specimens were investigated in the same manner, mRNA expression of COX-2 was quantitatively measured by real-time reverse transcription-polymerase chain reaction （RT-PCR） from 18 PLL specimens of patients with CSM and 18 PLL specimens of autopsy cases. Immunohistochemistry was used to evaluate the cellular location of COX-2 in PLL. Results A distinct amount of fibrotic area, chondrometaplastic tissue and calcification were found in the PLL of the patient group, compared with the control group. Type II collagen was apparent around chondrometaplastic cells. Ki-67 positive reaction was less than 5%. A COX-2 positive reaction was found in 9 of the patient specimens （39.1%） in which the COX-2 was released from vascular endothelial cells in the PLL. However, such reactions were not found in the control group. Real-time PCR showed that the mRNA expression level of COX-2 in the patient group was significantly higher than that in the control group （P〈0.01）. Conclusions Chondrometaplastic tissue producing type II collagen was identified as the most predominant pathological feature in the degenerative PLL. The higher expression of COX-2 might be related to degeneration of the PLL in CSM.

Effects of Electroacupuncture Intervention on Expression of Cyclooxygenase 2 and Microglia in Spinal Cord in Rat Model of Neuropathic Pain

Objective： To investigate the effect of electroacupuncture（EA） treatment on the expression of cyclooxygenase（COX） 2 and microglia in spinal cord by using rat model of neuropathic pain, and to probe into the relationship between COX 2 and microglia. Methods： The rats were randomly divided into 6 groups, including normal control group, model group, sham group, EA 1 group（distant acupoints ＋ local acupoints）, EA 2 group（local acupoints）, and EA 3 group（distant acupoints）. Thermal withdrawal latencies were evaluated at 1 day preoperatively and 3, 5 and 7 days postoperatively. At 7 days postoperatively, the spinal COX 2 m RNA was detected by reverse-transcription polymerase chain reaction. Double immunofluorescent staining technology was applied to screen and verify the relationship between altered COX 2 and microglia. Results： Compared with the model group, thermal withdrawal latencies increased after EA treatment（P〈0.01）. The expressions of COX 2 m RNA were up-regulated in spinal cord of rat on day 7 after surgery（P〈0.05）. Compared with the model group, EA stimulation（EA 1 and EA 2 groups） reversed the up-regulation of COX 2 m RNA expression（P〈0.05）. EA 1 and EA 2 groups might have better treatment effect compared with the EA 3 group. Fluorescent images displayed COX 2 and microglia expressed at common areas. Conclusions： EA was effective in analgesic and anti-inflammatory. EA has decreased the expression of spinal COX 2 m RNA in the trend of the therapeutic effect of ＂distant acupoints ＋ local acupoints＂, and ＂local acupoints＂ intervention may be superior to that of ＂distant acupoints＂ intervention. Microglia may be related to the formation of COX 2.

Comparative analysis of clinicopathological correlations of cyclooxygenase-2 expression in resectable pancreatic cancer

AIM:To perform a comparative analysis of clinicopathological correlations of cyclooxygenase2 (COX2) expression in pancreatic cancer, examined by monoclonal and polyclonal antibodies.METHODS: The COX2 expression in 85 resection specimens of pancreatic ductal adenocarcinoma was immunohistochemically examined using both monoclonal and polyclonal antibodies. The final immunoscores were obtained by multiplying the percentage of positive cells with the numeric score reflecting the staining intensity.COX2 expression levels were classified into three categories (0, 1+, and 2+) and the clinicopathological correlations were statistically evaluated and analyzed.RESULTS: The positive tumor expression rates of COX2 were 80.5% using monoclonal antibody and 69.4% using polyclonal antibody. In the KaplanMeier analysis, no significant correlations were found between levels of COX2 expression and overall survival (OS), but trends to longer OS were found in COX2 negative cases using monoclonal antibody. Significantly longer disease free survival was revealed in COX2 negative cases using monoclonal antibody (P = 0.019). No correlations between COX2 expression levels and grade (G), tumor (T) status and nodal (N) status were demonstrated. Low histological grade showed a strong association with a longer OS (P < 0.001). Correlation of survival and T status revealed a shorter OS in T3 tumors, but the results reached only marginal statistical significance (P = 0.070). In the multivariate Cox proportional hazards regression model, histological grade, T and N status remained valuable predictors of a worse survival with borderline significance for T [hazards ratio (HR) = 4.18 for G (if G = 3, P < 0.001); HR = 1.64 for T (if T = 3, P = 0.065); HR = 2.53 for N (if N = 1, P = 0.006)]. Higher grade, T or N status was associated with a worse OS. CONCLUSION: The immunohistochemically assessed level of COX2 expression does not seem to represent a valuable independent prognostic factor and is not superior to the conventional prognostic factors.

Extract of buckwheat sprouts scavenges oxidation and inhibits pro-inflammatory mediators in lipopolysaccharide-stimulated macrophages (RAW264.7)

OBJECTIVE： Buckwheat has been considered as a potential source of nutraceutical components on the world market of probiotic foodstuffs. The purpose of this study was to evaluate the effects of tartary buckwheat （Fagopyrum tataricum） sprouts on oxidation and pro-inflammatory mediators. METHODS： The anti-oxidant effects of buckwheat extract （BWE） and rutin were evaluated by using 1,1-diphenyl-2-picrylhydrazyl （DPPH）-and nitric oxide （NO）-scavenging activities, serum peroxidation and chelating assays. Lipopolysaccharide （LPS）-stimulated RAW264.7 cells were used to evaluate anti-inflammatory activities of buckwheat and rutin. NO production in LPS- stimulated RAW264.7 cells was determined by using Griess reagent. The expressions of inducible nitric oxide synthase （iNOS）, cyclooxygenase-2 （COX-2）, nuclear factor-kappa B （NF-κB） p65 subunit in cytosolic and nuclear portions were determined by Western blot analysis. Also, the production of inflammatory cytokines interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） was determined by enzyme-linked immunosorbent assay. RESULTS： Inhibitory concentration 50 values for DPPH- and NO-scavenging activities of BWE were 24.97 and 72.54 μg/mL respectively. BWE inhibited serum oxidation and possessed chelating activity. Furthermore, BWE inhibited IL-6 and TNF-a production in LPS-stimulated RAW264.7 cells. Also, BWE inhibited iNOS and COX-2 expression and NF-KB p65 translocation. CONCLUSION： Buckwheat sprouts possessed strong antioxidant activity and inhibited production of pro-inflammatory mediators in the applied model systems. Thus, buckwheat can be suggested to be beneficial in inflammatory diseases by inhibiting the free radicals and inflammatory mediators.

COX-2 in liver,from regeneration to hepatocarcinogenesis:What we have learned from animal models?

The use of animals lacking genes or expressing genes under the control of cell-specific promoters has signifi cantly increased our knowledge of the genetic and molecular basis of physiopathology,allowing testing of functional hypotheses and validation of biochemical and pharmacologic approaches in order to understand cell function.However,with unexpected frequency,gene knockout animals and,more commonly,animal models of transgenesis give experimental support to even opposite conclusions on gene function.Here we summarize what we learned on the role of cyclooxygenase 2(COX-2) in liver and revise the results obtained in 3 independent models of mice expressing a COX-2 transgene specifi cally in the hepatocyte.Upon challenge with pro-inflammatory stimuli,the animals behave very differently,some transgenic models having a protective effect but others enhancing the injury.In addition,one transgene exerts differential effects on normal liver physiology depending on the transgenic animal model used.

Thymoquinone suppresses migration of Lo Vo human colon cancer cells by reducing prostaglandin E2 induced COX-2 activation

AIM To identify potential anti-cancer constituents in natural extracts that inhibit cancer cell growth and migration.METHODS Our experiments used high dose thymoquinone(TQ) as an inhibitor to arrest Lo Vo(a human colon adenocarcinoma cell line) cancer cell growth, which was detected by cell proliferation assay and immunoblotting assay. Low dose TQ did not significantly reduce Lo Vo cancer cell growth. Cyclooxygenase 2(COX-2) is an enzyme that is involved in the conversion of arachidonic acid into prostaglandin E2(PGE2) in humans. PGE2 can promote COX-2 protein expression and tumor cell proliferation and was used as a control.RESULTS Our results showed that 20 μmol/L TQ significantly reduced human LoV o colon cancer cell proliferation. TQ treatment reduced the levels of p-PI3 K, p-Akt, p-GSK3β, and β-catenin and thereby inhibited the downstream COX-2 expression. Results also showed that the reduction in COX-2 expression resulted in a reduction in PGE2 levels and the suppression of EP2 and EP4 activation. Further analysis showed that TG treatment inhibited the nuclear translocation of β-catenin in LoV o cancer cells. The levels of the cofactors LEF-1 and TCF-4 were also decreased in the nucleus following TQ treatment in a dose-dependent manner. Treatment with low dose TQ inhibited the COX-2 expression at the transcriptional level and the regulation of COX-2 expression efficiently reduced LoV o cell migration. The results were further verified in vivo by confirming the effects of TQ and/or PGE2 using tumor xenografts in nude mice.CONCLUSION TQ inhibits LoV o cancer cell growth and migration, and this result highlights the therapeutic advantage of using TQ in combination therapy against colorectal cancer.

环氧化酶-2和血管内皮生长因子C表达与喉癌淋巴管转移

环氧化酶-2(cyclooxygenase-2,COX-2)是前列腺素合成的关键酶,在头颈鳞状细胞癌(简称鳞癌)等多种恶性肿瘤中表达上调,并与肿瘤发生和发展有关。淋巴管转移是喉鳞癌最常见的转移方式,血管内皮生长因子C(vascular endothelial growth factor C,VEGF-C)

环氧化酶-2和Survivin mRNA在舌癌中的表达及临床意义

1资料与方法 1.1一般资料。收集我科2006～2011年舌癌手术切除标本60例,其中男37例,女23例,年龄40～75岁,所有患者手术前均未经化疗、放疗或者免疫治疗。另收集舌外伤、舌修整正常舌黏膜组织30例,全部标本1份经甲醛溶液固定,用于病理诊断,另1份置液氮中冻存备做逆转录聚合酶链式反应（RT-PCR）。

Influence of L-arginine on the Expression of eNOS and COX2 in Experimental Pulmonary Thromboembolism

The influence of L-arginine on endothelial nitric oxide synthase （eNOS） and cyclooxygenase 2 （COX2） was observed in experimental pulmonary thromboembolism and the action mechanism on pulmonary thromboembolism was explored. Wistar rats were randomly divided into control group, model group and treatment group. Pulmonary thromboembolism models were established by auto-blood back transfusion, and L-Arg 100 mg/kg was intraperitoneally injected after successful model preparation. The animals were sacrificed at 3 h, 1 day, 3 days and 7 days after embolism. Plasma NO, TXB2 and 6-Keto-PGF1 α were detected. The expression of eNOS and COX2 protein and mRNA in pulmonary tissues was detected by immunohistochemistry and RT-PCR respectively. The results showed that pulmonary thrombosis could be seen post pulmonary embolism and inflammatory reaction was significant. Plasma NO was decreased （P〈0.01）, and the levels of TXB2, 6-Keto-PGF1α and T/P ratio were all elevated. The expression of eNOS protein and mRNA in the pulmonary tissue was down-regulated （P〈0.05）, while that of COX2 protein and mRNA was upregulated （P〈0.01）. In treatment group, the level of NO was increased, the levels of TXB2 and T/P ratio were decreased, but the level of 6-Keto-PGF1 α was increased. The expression of eNOS protein and mRNA in pulmonary tissue was upregulated （P〈0.05）, while that of COX2 protein and mRNA was down-regulated （P〈0.05）. In conclusion, L-arginine can educe the role of pulmonary tissue protection through up-regulating the expression of intra-pulmonary NOS and down -regulating COX2 in pulmonary thromboembolism.

塞来昔布对鼻咽癌放射治疗增敏作用的临床研究

我们从2006年1月～2008年1月在鼻咽癌放射治疗中,应用塞来昔布配合增敏作用,报道如下。1资料与方法1.1一般资料。鼻咽癌初诊患者64例,男44例,女20例,年龄28～68岁。Karnofsky评分70分以上。临床分期采用1992年福州分期标准,血、尿常规、肝肾功能、心电图均正常,近1个月内未进行抗肿瘤药物治疗。

The Effects of Nimesulide Combined with Cisplatin on Lung Cancer

To study the effects of cyclooxygenase 2 selective inhibitor Nimesulide (NIM) combined with Cisplatin (DDP) on human lung cancer and the possible mechanisms, the proliferation and apoptosis of human lung cancer cell line A549 were evaluated by MTT reduction assay and flow cytometry respectively. The inhibitory effect on neoplasia in vivo was tested on nude mice subcutaneously implanted tumor. Our results showed that NIM and DDP could inhibit A549 cell proliferation in a concentration-dependent pattern; this action was enhanced when NIM (25 μmol/L) was given in combination with DDP and they worked in a synergistic or additive pattern as DDP concentration ≥1 μg/ml. NIM and DDP could induce A549 cells apoptosis and the action was augmented when used in combination (P<0.01). NIM and DDP could inhibit the growth of subcutaneously implanted tumors on nude mice (P<0.05,P<0.01) and the inhibitory rate of NIM combined with DDP was significantly higher than that of NIM or DDP group (P<0.01, P<0.01). It is concluded that combined use of NIM and DDP has significant synergistic antitumor effects on lung cancer cell line A549 and in animals in vivo. The synergy may be achieved by growth inhibition and apoptosis induction.

Component Analysis of Shuanghuanglian Freeze-dried Powder and Its Effects on Human Hepatocyte Function when Combined with Cefradine

[Objectives]To detect the contents of components in Shuanghuanglian freeze-dried powder and to explore the effects of Shuanghuanglian freeze-dried powder and its components on human hepatocytes(HL-7702)alone or in combination with cefradine.[Methods]High performance liquid chromatography(HPLC)was used to detect the contents of baicalin,wogonin,chlorogenic acid and forsythin,the main components of Shuanghuanglian freeze-dried powder.HL-7702 cells were cultured with Shuanghuanglian freeze-dried powder and the main components of Shuanghuanglian freeze-dried powder alone or in combination with cefradine.Enzyme linked immunosorbent assay(ELISA)was used to detect the contents of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the cell supernatant after culture,and HPLC was used to detect the expression level of adenosine diphosphate(ADP)and adenosine triphosphate(ATP);agarose gel electrophoresis was used to detect the expression of cyclooxygenase 2(COX-2)and heme oxygenase-1(HO-1)in HL-7702 cells.[Results]In Shuanghuanglian freeze-dried powder for injection,the content of baicalin was the highest,and the content of wogonin was the lowest.Compared with the control group,the expressions of AST and ALT in human hepatocytes(HL-7702)in high-dose baicalin group,forsythin group and wogonin group decreased(P<0.05,P<0.01),while the expression of ALT in chlorogenic acid+cefradine group(0.046 mg/mL)and forsythin+cefradine group(0.046 mg/mL)increased(P<0.05,P<0.01),and the expression of AST had no significant difference(P>0.05);the results in the low-dose group were similar to those in the high-dose group.Compared with the control group,ATP expression in chlorogenic acid group,chlorogenic acid+cefradine group(0.046 mg/mL)and forsythin+cefradine group(0.046 mg/mL)in the high-dose group decreased(P<0.05,P<0.01),and ADP expression was not significantly different(P>0.05);in the low-dose group,the expression of ATP and ADP increased in baicalin group(P<0.05),but decreased in wogonin group,baicalin+cefradine group(0.046 mg/mL)and wogonin group+cefradine group(0.046 mg/mL)(P<0.05,P<0.01).Compared with the control group,the expressions of COX-2 and HO-1 in HL-7702 cells in the cefradine group showed no significant difference(P>0.05).The expression of HO-1 and COX-2 in the different dose groups of Shuanghuanglian and the group combined with cefradine increased,and the difference was significant(P<0.05,P<0.01).[Conclusions]The components of Shuanghuanglian freeze-dried powder for injection had effects on hepatocytes,of which baicalin had a significant effect,and the effect of cefradine on hepatocytes was increased when used in combination with cefradine.

Efficacy and safety of perioperative parecoxib for acute postoperative pain treatment in children： a meta-analysis

Perioperative parecoxib administration reduces postoperative pain, opioid consumption, and adverse events in adult patients. However, the efficacy and safety of parecoxib in children remain unclear. This meta-analysis included related published studies to address this concern. Eight databases in the literature until February 2015 were systematically explored to identify randomized controlled trials （RCTs） comparing perioperative parecoxib administration and placebo/standard treatments for acute postoperative pain in children. Primary outcomes were postoperative pain scores and adverse events. The Face, Legs, Activity, Crying, Consolability scale was used to score pain in children younger than 6 years, whereas the Visual Analog Scale was used in children older than 6 years. Secondary outcomes were sedation scores （measured using the Ramsay scale）, agitation scores （measured using the Sedation-Agitation Scale）, and opioid consumption. The methodological quality of RCTs was independently assessed in accordance with the ＂Risk of bias＂ of Cochrane Collaboration. Data were analyzed using Review Manager 5.2. Twelve RCTs involving 994 patients met the inclusion criteria. Compared with children who received placebo treatment, those who received parecoxib demonstrated lower early （2 h） and later （12 h） postoperative pain scores; lower incidence rates of postoperative nausea, vomiting, and agitation; higher early （1 h） postoperative sedation scores; and lower agitation scores. Similarly, children who received parecoxib had lower early （2 h） and later （12 h） postoperative pain scores, lower incidence rates of postoperative nausea and vomiting, and lower early （1 h） postoperative sedation scores compared with those who received standard treatments; however, these children showed no significant difference in agitation scores. Unfortunately, data on the effect of parecoxib on opioid consumption were insufficient. Overall, these results suggested that perioperative parecoxib administration was associated with less acute postoperative pain and fewer adverse events compared with placebo or standard treatments. Parecoxib administration also resulted in less emergence agitation compared with placebo treatment and less excessive sedation concern compared with standard treatments. However, the long-term effects, effects on opioid consumption, and patient satisfaction of parecoxib administration warrant further investigation.

Anti-helicobacter pylori effect of total alkaloids of sophora alopecuroides in vivo

Background Helicobacterpylori （H.pylori） infection could lead to most gastroduodenal diseases and is even identified as a carcinogen of gastric cancer.Total alkaloids of sophora alopecuroides （TASA） is widely used in herbal remedies to treat various infectious diseases,including stomach-associated diseases.This study is aimed at evaluating the antimicrobial activity of TASA on H.pylori-infected BALB/c mice mouse gastritis.Methods Totally 120 BALB/c mice were orally inoculated with H.pylori Bacterial liquid to construct BALB/c mice H.pylori infection gastritis animal model,after the model was successfully created.We randomly assigned 100 infected mice into 10 treatment groups,the first group （normal saline）; the second group （bismuth pectin）; the third group （omeprazole）; the fourth group （TASA 2 mg/d）; the fifth group （TASA 4 mg/d）; the sixth group （TASA 5 mg/d）; the seventh group （TASA ＋ bismuth pectin）; the eighth group （TASA ＋ omeprazole）; the ninth group （bismuth pectin ＋ clarithromycin ＋ metronidazole）;the tenth group （omeprazole ＋ clarithromycin ＋ metronidazole）,5 other non-infected mice as negative control.Mice were orally inoculated twice a day and 7 days continuously.Then the mice were killed 4 weeks after treatment,we used realtime PCR to detect 16sDNA of H.pylori to test both the colonization and the clearance mice of bacteria of each treatment.We applied hematoxylin and eosin （HE） staining and immunostaining of mice gastric mucosa to observe the general inflammation and related factors interleukin 8 （IL-8）,cyclooxygenase 2 （COX-2）,and nuclear factor-kappa B （NF-KB） expression change after treatments.Results Firstly,we ensured that after 6-week intragastric administration,the bacteria colonization reached an exceed peak which is far higher than positive threshold （P ＜0.001）; secondly,after treatments,it is revealed that TASA combined with omeprazole or bismuth pectin showed promising antimicrobial activity against H.pylori as well as conventional triple therapy （P ＜0.001）; thirdly,HE staining showed that the inflammation on mice gastric mucosal membrane were also relieved obviously in TASA combined treatments and conventional triple therapy compared with normal saline treated mice,moreover,from immunohistochemistry results,H.pylori-induced IL-8,COX-2,and NF-KB were consistently suppressed in seventh,eighth,ninth,and tenth group to a certain extent.Conclusion These results open the possibility of taking TASA as an anti-inflammatory agent for H.pylori gastritis.

Efficacy of horse oil on lipopolysaccharide-induced inflammation in human keratinocytes

OBJECTIVE: To investigate the efficacy of horse oil on lipopolysaccharide(LPS)-induced inflammation in human keratinocytes.METHODS: Western blot analysis was performed to measure the expression of cyclooxygenase-2(COX-2) and IκBα. ELISA was used to analyze prostaglandin E2(PGE2) levels.RESULTS: Horse oil decreased LPS-induced COX-2 and PGE2 levels in a dose-dependent manner. Nuclear factor-kappa B(NF-κB) plays a key role in the expression of inflammatory cytokines and mediators. Therefore, we investigated the influence of horse oil on the NF-κB signaling pathways. Horse oil inhibited translocation of NF-κB from the cytosol to the nucleus. Furthermore, LPS-induced degradation of IκBα was recovered by horse oil. The activation of p38 mitogen-activated protein kinase(MAPK) reportedly induces degradation of IκBα In agreement with this, LPS activated p38 MAPK and caused IκBα degradation. Conversely, horse oil inhibited LPS-induced p38 MAPK activation and IκBαdegradation. In addition, a specific p38 MAPK inhibitor, SB203580, blocked IκBα degradation.CONCLUSION: Horse oil decreased COX-2 and PGE2 by inhibiting p38 MAPK activation, IκBα degradation, and the translocation of NF-κB.