Changes and their significances of IFN-γ, TNF-α and IL-2 level in mouse peripheral blood following intraperitoneal injection with influenza viruses

Objective To observe changes of IFN-γ, TNF-α and IL-2 levels in mouse peripheral blood following intraperitoneal injection with influenza viruses. Methods Mice were divided into 3 groups randomly and injected with human H3N2 influenza virus, avian H9N2 influenza virus and sterilized virus-free allantoic fluid, respectively. Sera in different groups were collected at several time points after virus injection, and the levels of IFN-γ, TNF-α and IL-2 in peripheral blood were tested by sandwich ELISA. Statistical analysis was made using analysis of variance and LSD-t test. Results The levels of IFN-γ and IL-2 in peripheral blood in influenza viruses injection groups were significantly higher than those in negative control group, and no significant difference in TNF-α level was found between influenza viruses injection groups and negative control group. Conclusion Intraperitoneal injection with human or avian influenza viruses can promote the production of IFN-γ and IL-2 in mouse peripheral blood, but it has little effect on the production of TNF-α.     

核心蛋白聚糖协同白细胞介素24对人外周血单个核细胞作用的体外研究

Objective To investigate the co-effect of decorin (DCN) and interleukin-24 (IL-24) on proliferation and Interferon-γ (IFN-γ) secretion of human peripheral blood mononuclear cells (PBMCs). Methods Recombinant plasmid pcDNA3. 1 (+)-DCN and pcDNA3. 1 (+)-IL-24 were constructed and transfected into PBMCs by liposome. After transfected with different plasmids, the PBMCs were divided into 6 groups;blank control group, Lipofectamine TM group, empty vector group, DCN group, IL-24 group, DCN and IL-24 group. PBMC proliferation was determined by methyl thiazolyl tetrazolium assay. IFN-7 expression in the supernatant was detected by ELISA. Flow cytometry was used to determine the surface expression of programmed death-1 (PD-1) on PBMCs. Statistical analysis was made using LSD-t test. Results The recombinant plasmid pcDNA3. 1 (+)-IL-24 was constructed successfully. PBMC proliferation and IFN-7 secretion were significantly higher in DCN and IL-24 group, and the expression of PD-1 was also upregulated obviously. Conclusion The combination of DCN and IL-24 could promote PBMC proliferation and strengthen the function of PBMCs in vitro.     

核心蛋白聚糖协同白细胞介素24对人外周血单个核细胞作用的体外研究

Objective To investigate the co-effect of decorin (DCN) and interleukin-24 (IL-24) on proliferation and Interferon-γ (IFN-γ) secretion of human peripheral blood mononuclear cells (PBMCs). Methods Recombinant plasmid pcDNA3. 1 (+)-DCN and pcDNA3. 1 (+)-IL-24 were constructed and transfected into PBMCs by liposome. After transfected with different plasmids, the PBMCs were divided into 6 groups;blank control group, Lipofectamine TM group, empty vector group, DCN group, IL-24 group, DCN and IL-24 group. PBMC proliferation was determined by methyl thiazolyl tetrazolium assay. IFN-7 expression in the supernatant was detected by ELISA. Flow cytometry was used to determine the surface expression of programmed death-1 (PD-1) on PBMCs. Statistical analysis was made using LSD-t test. Results The recombinant plasmid pcDNA3. 1 (+)-IL-24 was constructed successfully. PBMC proliferation and IFN-7 secretion were significantly higher in DCN and IL-24 group, and the expression of PD-1 was also upregulated obviously. Conclusion The combination of DCN and IL-24 could promote PBMC proliferation and strengthen the function of PBMCs in vitro.     

Effects of methione-enkaphalin on interleukin- 1, tumor necrosis factor-α and nitric oxide production from rat microglia stimulated with lipopolysaccharides

Methione-enkaphine (Met-Enk), an opioid pentapeptide with awide distribution in the central nervous system and peripheral tissue, was studied its effects on proinflamamtory cytokines, such as, interleukin-I (IL-1) and tumor necrosis factor-α(TNF-α) and nitric oxide (NO) which are involved in various physiopathological conditions in the CNS on rat microglia cultures. The biological activities of IL-1, TNF-α was tested by mouse thymocyte proliferation and L929 cytotoxity assay.NO levels were represented as nitrite concentration which is determined     

Mechanism study of OVA-induced allergic asthma in mice based on lipid metabolomics北大核心CSCD

Aim To investigate the mechanism and search for potential biomarkers of ovalbumin ( OVA ) -induced asthma in mice base on lipidomics. Methods A BALB/c mouse model of asthma was prepared by OVA. TNF-α, IL-4, IL-10, IFN-γ levels in BALF and IgE level in serum were measured by ELISA. The inflammatory changes in mouse lung tissue were observed using HE staining. Lipid mediators ( LMs) in lung tissue and serum were quantified with UPLC-MS/ MS strategy. Results IgE level in serum and TNF-α, IFN-γ levels in BALF were higher (P <0.05) of asthmatic mice.Typical inflammatory manifestations were seen in lung tissue of asthmatic mice. A total of 57 lipid mediators were quantified with UPLC-MRM. LMs metabolic profiles differed significantly in serum and lung tissue between asthmatic and normal mice, 17 significantly different LMs were found in lung tissue and 6 LMs were found in serum, and the differential metabolites were produced through the cyclooxygenase (COX), lipoxygenase (LOX) and cytochrome P450 oxidase (P450) metabolic pathways. Conclusions OVA-induced allergic asthma can cause disorder of lip-id mediators, LMs and cytokines are involved in the occurrence and development of asthma. The differential LMs have potential research value as biomarkers for the development of allergic asthma. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

康艾注射液对人结直肠癌Lovo细胞Th1/Th2类细胞因子的影响

Objective To explore the effect of Kang'ai injection on Th1/Th2 cytokine in human colon carcinoma cell line Lovo.Methods Lovo cells were cultured by conventional ways until they were in exponential growth phase and then divided into 4 groups:one control group and three experimental groups with different concentration of Kang'ai injection.After 72 hours culture,Th1 type cytokines (IL-2,IFN-γ) and Th2.type cytokines (IL-4,IL-10) in the culture supernatant of Lovo cells in each group were separately detected by double-antibody sandwich enzyme-linked immunosorhent assay (ELISA).Results After the coculture of Lovo cells and Kang'ai injection,Th2 type cytokine levels in the supernatants were significant lower than the control group (P＜0.05),and Th1 type cytokines in each group were of little change.Conclusions Kang'ai injection markedly downregulated Th2 type cytokines in Lovo cells,and contributed to the switching from Th2 to Th1.     

Plasma thromboxane B2 and 6-keto-prostaglandin-F1α in patients with type 2 diabetes mellitus

Objective To investigate the changes of plasma thrombexane B2 (TXB2) and 6-keto-prostaglan-din-F1α (6-keto-PGF1α) in patients with type 2 diabetic mellitus and their roles in the pathogenesis of diabetic macroangiopathy. Methods Plasma levels of TXB2 and 6-keto-PGF1α were measured with RIA in 60 cases of type 2 diabetic patients(43 patients with and 17 without diabetic macroangiopathy), with 31 cases of controls. Results Plasma levels of TXB2 in type 2 diabetic patients were significantly higher than those in controls. However, plasma levels of 6-keto-PGF1α in type 2 diabetic patients were significantly lower than those in controls. Changes of both indexes were more significant in type 2 diabetic patients with macroangiopathy than those without macroangiopathy. Plasma levels of TXB2 showed negative correlation with 6-keto-PGF 1α. Conclusion TXB2 and 6-keto-PGF 1 α may play important roles in the pathogenesis of diabetic macroangiopathy. Determination of TXB2 and 6-keto-PGF1α levels in type 2 diabetic patients may be useful for early detection of diabetic macroangiopathy.     

Plasma thromboxane B2 and 6-keto-prostaglandin-F1α in patients with type 2 diabetes mellitus

Objective To investigate the changes of plasma thrombexane B2 (TXB2) and 6-keto-prostaglan-din-F1α (6-keto-PGF1α) in patients with type 2 diabetic mellitus and their roles in the pathogenesis of diabetic macroangiopathy. Methods Plasma levels of TXB2 and 6-keto-PGF1α were measured with RIA in 60 cases of type 2 diabetic patients(43 patients with and 17 without diabetic macroangiopathy), with 31 cases of controls. Results Plasma levels of TXB2 in type 2 diabetic patients were significantly higher than those in controls. However, plasma levels of 6-keto-PGF1α in type 2 diabetic patients were significantly lower than those in controls. Changes of both indexes were more significant in type 2 diabetic patients with macroangiopathy than those without macroangiopathy. Plasma levels of TXB2 showed negative correlation with 6-keto-PGF 1α. Conclusion TXB2 and 6-keto-PGF 1 α may play important roles in the pathogenesis of diabetic macroangiopathy. Determination of TXB2 and 6-keto-PGF1α levels in type 2 diabetic patients may be useful for early detection of diabetic macroangiopathy.     

Plasma thromboxane B2 and 6-keto-prostaglandin-F1α in patients with type 2 diabetes mellitus

Objective To investigate the changes of plasma thrombexane B2 (TXB2) and 6-keto-prostaglan-din-F1α (6-keto-PGF1α) in patients with type 2 diabetic mellitus and their roles in the pathogenesis of diabetic macroangiopathy. Methods Plasma levels of TXB2 and 6-keto-PGF1α were measured with RIA in 60 cases of type 2 diabetic patients(43 patients with and 17 without diabetic macroangiopathy), with 31 cases of controls. Results Plasma levels of TXB2 in type 2 diabetic patients were significantly higher than those in controls. However, plasma levels of 6-keto-PGF1α in type 2 diabetic patients were significantly lower than those in controls. Changes of both indexes were more significant in type 2 diabetic patients with macroangiopathy than those without macroangiopathy. Plasma levels of TXB2 showed negative correlation with 6-keto-PGF 1α. Conclusion TXB2 and 6-keto-PGF 1 α may play important roles in the pathogenesis of diabetic macroangiopathy. Determination of TXB2 and 6-keto-PGF1α levels in type 2 diabetic patients may be useful for early detection of diabetic macroangiopathy.     

Plasma thromboxane B2 and 6-keto-prostaglandin-F1α in patients with type 2 diabetes mellitus

Objective To investigate the changes of plasma thrombexane B2 (TXB2) and 6-keto-prostaglan-din-F1α (6-keto-PGF1α) in patients with type 2 diabetic mellitus and their roles in the pathogenesis of diabetic macroangiopathy. Methods Plasma levels of TXB2 and 6-keto-PGF1α were measured with RIA in 60 cases of type 2 diabetic patients(43 patients with and 17 without diabetic macroangiopathy), with 31 cases of controls. Results Plasma levels of TXB2 in type 2 diabetic patients were significantly higher than those in controls. However, plasma levels of 6-keto-PGF1α in type 2 diabetic patients were significantly lower than those in controls. Changes of both indexes were more significant in type 2 diabetic patients with macroangiopathy than those without macroangiopathy. Plasma levels of TXB2 showed negative correlation with 6-keto-PGF 1α. Conclusion TXB2 and 6-keto-PGF 1 α may play important roles in the pathogenesis of diabetic macroangiopathy. Determination of TXB2 and 6-keto-PGF1α levels in type 2 diabetic patients may be useful for early detection of diabetic macroangiopathy.     

Study on mechanisms of hypertension in rat adult offspring following prenatal exposure to immuno-inflammatory stimulants

Objective Essential hypertension(EH)is one of the most common cardiovascular disease and the main causes of human fatility.Recently significant progress has been made in our lab,it was found that exterior stimulation during pregnancy may play a key role in chronicle adult disease.However,what factors affect the growth of fetus after those exterior stimulation and why has not been reported.Based on our previous finding,this study intends to investigate how immuno-inflammatory stimulation affect the development of embryo.Methods 1.Sprague-Dawley(SD)rats,dams in each group received i.p.injections of 0.79 mg·kg-1 LPS,8 mg·kg-1 zymosan or sterile saline respectively on their gestational days 8,10,and 12.2.The serums were collected in tail nick at 2 h after the last injection,and the amniotic fluid was mixed at 2,12,24,48 h after the last injection.TNF-α and IL-6 levels of serum and amniotic fluid were measured by RIA method.3.TNF-α and IL-6 mRNA levels were quantitated in amnion,placenta,amniotic fluid,Embryo and macrophage by real-time fluorescent quantitative-PCR.Results 1.The serum level of TNF-α and IL-6 in LPS group and zymosan group was higher than that in control group(P<0.01).It showed that there was immuno-imflammatory response after LPS or zymosan injection in rats.The mRNA levels of TNF-α and IL-6 was very higher in macrophage than in other organization.2.In embryo,the mRNA level of IL-6 was more than other organization,but the mRNA level of TNF-α was lower than other organization.However,the IL-6 mRNA level of LPS group and zymosan group was higher several dozens times than control group on 24 h and 48 h.Conclusions It suggested that IL-6 was important in the model that prenatalexposure to immuno-inflammatory stimulant results in increases of blood pressure and body weight in rats.     

The dynamic changes of serum tumor necrosis factor-α and the clinical significance in acute lung injury rats caused by mechanical trauma

Objective To determine the dynamic changes and mechanism of tumor necrosis factor-α(TNF-α) in serum in acute lung injury(ALI) induced by mechanical trauma in mechanical trauma rat model. Methods Totally 40 male Wistar rats were randomly divided into sham group and trauma group, sham-acute lung injury group and acute lung injury group. Noble-Collip drum was used to establish mechanical trauma rat model. Intraperitoneal injection of TNF-α established acute lung injury model. All rats after modeling of abdominal aortic blood sampling time points were killed and the serum levels of TNF-α were asasyed by ELISA. Results Serum TNF-α levels (334. 78 ±± 28) ng/L in the trauma group were significantly higher than those in the sham group( 177 ±10 ) ng/L (P ＜0. 01 ). The pathological results showed that both in trauma group and acute lung injury group lung infection were very obvious, while it was basically normal in both sham group and sham-acute lung injury group. Conclusions Serum TNF-αt levels of mechanical trauma may be associated with acute lung injury after mechanical trauma. Therefore, dynamic observation of the change of serum TNF-α after mechanical trauma will help the assessment and prognosis of the disease, it has important clinical significance.     

The dynamic changes of serum tumor necrosis factor-α and the clinical significance in acute lung injury rats caused by mechanical trauma

Objective To determine the dynamic changes and mechanism of tumor necrosis factor-α(TNF-α) in serum in acute lung injury(ALI) induced by mechanical trauma in mechanical trauma rat model. Methods Totally 40 male Wistar rats were randomly divided into sham group and trauma group, sham-acute lung injury group and acute lung injury group. Noble-Collip drum was used to establish mechanical trauma rat model. Intraperitoneal injection of TNF-α established acute lung injury model. All rats after modeling of abdominal aortic blood sampling time points were killed and the serum levels of TNF-α were asasyed by ELISA. Results Serum TNF-α levels (334. 78 ±± 28) ng/L in the trauma group were significantly higher than those in the sham group( 177 ±10 ) ng/L (P ＜0. 01 ). The pathological results showed that both in trauma group and acute lung injury group lung infection were very obvious, while it was basically normal in both sham group and sham-acute lung injury group. Conclusions Serum TNF-αt levels of mechanical trauma may be associated with acute lung injury after mechanical trauma. Therefore, dynamic observation of the change of serum TNF-α after mechanical trauma will help the assessment and prognosis of the disease, it has important clinical significance.     

细胞因子与2型糖尿病胰岛素抵抗的相关性研究

Objective To study on the changes of the IL-6、IL-8、TNF-α、CRP in the type 2 diabetes with insulin resistance and the relation of plasma glucose or insulin resistance and cytokines. Methods (1) To assay the serum level of cytokines in 125 cases with type 2 diabetes mellitus(T2DM) and 40 normal controls. (2)To measure the level of fasting plasma glucose (FPG), hemoglobin A1c (HBA1c), fasting insulin (FINS), fasting C-peptide (FCP). (3)To analyze the relation of the level of FPG、HBA1c、FINS、FCP and cytokines. Results (1)The serum levels of cytokines in T2DM group were significantly higher than in control groups, IL-6、CRP、TNF-α in T2DM group with insulin resistance were significantly increase than in T2DM group without insulin resistance; (2)There was significantly positively correlation between insulin resistance and the increasing of cytokine levels. Conclusion There is an excess activation of cytokines in T2DM, which is significantly correlated with insulin resistance. Cyto-kines plays an important role in the occurrence and development of T2DM and insulin resistance.     

Mechanism of seahorse ameliorating depression-like behavioral and pathological changes caused by chronic stress in zebrafish北大核心CSCD

Aim To investigate the effect of marine herbal seahorse on chronic unpredictable mild stress ( CUMS ) -induced depression-like model in zebrafish. Methods Adult zebrafish were divided into control, Stress,Stress + low dose (Stress +0.044% SH) and Stress + high dose (Stress +0. 22% SH) seahorse intervention groups, and depression-like behavior was identified by novel tank test (NTT), cortisol, interleukin ( IL )-6 and interferon (IFN )-γ levels were detected by ELISA. The levels of dopamine (DA) ,norepinephrine (NE), 5-hydroxytryptamine (5-HT) and 5-hydroxyin-doleacetic acid (5-HIAA) were determined by high performance liquid chromatography. The mRNA expression levels of tryptophan hydroxylase(TPH)-2 and 5-HT2A receptor were measured by real-time quantitative PCR. Results Compared with the control group, the Stress group showed significantly longer latency to reach the top in NTT, significantly reduced number of transfers to the top region and top residence time, significantly increased levels of cortisol and IL-6, IFN-γ protein, significantly reduced levels of DA and 5-HT in brain as well as increased metabolism rate of 5-HT, while 5-HT2A mRNA expression was up-regulated and TPH2 mRNA expression was down-regulated. In contrast, low-dose seahorse intervention effectively reduced anxiety, decreased cortisol and IL-6 and IFN-γ concentrations, increased monoamine neurotransmitter levels and reversed dysregulation of the 5-HT ergic system in CUMS zebrafish. Conclusion Seahorse may exert an-tidepressant effects through anti-inflammation and mod¬ulation of monoamine neurotransmitter levels. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

上海地区人群甲型H1N1流感抗体水平监测分析

Objective To understand levels of antibody to swine-origin influenza A(H1 N1) virus (A/ H1N1) among different age groups in Shanghai, and to compare the levels between antibodies to A/H1N1 and to 2008-2009 seasonal influenza vaccine virus A/Brisbane/59/2007(H1N1). Methods The anti-A/HlN1 antibody levels were determined among 5 age groups (0-5 months, 6 months-4 years, 5-24 years, 25-59 years, and ≥60 years) with routine micro-hemagglutination inhibition test. The positive rates of the antibodies among different age groups were compared by Pearson's X2 test and Fisher' s exact probability test. Results The positive rate of antibody to A/H1N1 in the whole population in Shanghai was 9.2% (37/404). The highest rate was 25.0% (21/84) in the age group of ≥60 years, the next was 10. 0% (8/80) in the age group of 25-59 years, the positive rates in other groups were lower. The result of Pearson's X2 test showed that there were statistically significant differences in the positive rates of the antibodies among different age groups. Of 329 sera positive for antibody to 2008 -2009 seasonal influenza vaccine virus A/Brisbane/59/ 2007 (H1N1), 31 had anti-A/HlNl antibodies, and of 75 sera negative for antibody to A/Brisbane/59/ 2007(H1N1), 6 had anti-A/HlNl antibodies. The result of Pearson's X2 test showed that there were no significant differences in positive rates of antibody to A/H1N1 between the two types of population. Conclusions The anti-A/HlNl levels in Shanghai population are low, and people are generally susceptible to A/H1N1. The sera positive for antibody to A/Brisbane/59/2007(H1N1) can not provide cross-protection against A/H1N1.     

上海地区人群甲型H1N1流感抗体水平监测分析

Objective To understand levels of antibody to swine-origin influenza A(H1 N1) virus (A/ H1N1) among different age groups in Shanghai, and to compare the levels between antibodies to A/H1N1 and to 2008-2009 seasonal influenza vaccine virus A/Brisbane/59/2007(H1N1). Methods The anti-A/HlN1 antibody levels were determined among 5 age groups (0-5 months, 6 months-4 years, 5-24 years, 25-59 years, and ≥60 years) with routine micro-hemagglutination inhibition test. The positive rates of the antibodies among different age groups were compared by Pearson's X2 test and Fisher' s exact probability test. Results The positive rate of antibody to A/H1N1 in the whole population in Shanghai was 9.2% (37/404). The highest rate was 25.0% (21/84) in the age group of ≥60 years, the next was 10. 0% (8/80) in the age group of 25-59 years, the positive rates in other groups were lower. The result of Pearson's X2 test showed that there were statistically significant differences in the positive rates of the antibodies among different age groups. Of 329 sera positive for antibody to 2008 -2009 seasonal influenza vaccine virus A/Brisbane/59/ 2007 (H1N1), 31 had anti-A/HlNl antibodies, and of 75 sera negative for antibody to A/Brisbane/59/ 2007(H1N1), 6 had anti-A/HlNl antibodies. The result of Pearson's X2 test showed that there were no significant differences in positive rates of antibody to A/H1N1 between the two types of population. Conclusions The anti-A/HlNl levels in Shanghai population are low, and people are generally susceptible to A/H1N1. The sera positive for antibody to A/Brisbane/59/2007(H1N1) can not provide cross-protection against A/H1N1.     

血清白细胞介素及肿瘤坏死因子水平与急性脑梗死关系的研究

目的 探讨急性脑梗死患者血清白细胞介素6(IL-6)、IL-8和肿瘤坏死因子α(TNF-α)水平的变化及与急性脑梗死的关系.方法 选择急性脑梗死患者90例,按梗死体积及神经功能缺损程度进行分组,采用酶联免疫吸附法检测血清IL-6、IL-8和TNF-α水平并与30例对照组比较.结果 脑梗死组IL-6、IL-8和TNF-α水平[(158.4±25.8)ng/L、(168.3±18.8)ng/L、(2.9±1.0)μg/L]明显高于对照组[(68.8±5.8)ng/L、(62.3±7.5)ng/L、(0.8±0.2)μg/L],2组比较差异有统计学意义(P＜0.05).小梗死组、中梗死组、大梗死组患者血清IL-6、IL-8和TNF-α水平依次逐级升高,组间比较差异有统计学意义(P＜0.05);轻型组、中型组、重型组患者血清IL-6、IL-8和TNF-α水平逐渐升高,组间比较差异有统计学意义(P＜0.05).结论 血清IL-6、IL-8和TNF-α水平与脑梗死体积及临床神经功能缺损程度密切相关,检测IL-6、IL-8和TNF-α有助于急性脑梗死患者病情及预后的判断.

Abstract：

Objective To investigate the relationship among the serum levels of IL-6,IL-8,TNF-α and acute cerebral infarction (ACI). Methods Ninty patients with ACI were divided into different groups according to the size of infarction and clinical neurologic impairment degree score. The serum level of IL-6,IL-8 and TNF-αwere detected by enzyme-linked immunosorbent assay (ELISA). Results The serum IL-6,IL-8 and TNF-α levels in the patients with ACI within 72 hours after the onset were significantly higher than those in the healthy subjects (P＜0.05). The serum levels of IL-6,IL-8 and TNF-α increased with the volume of infarction and the differences between the groups were significant (all P＜0.05). The serum levels of IL-6,IL-8 and TNF-α increased with the clinical neurologic impairment degree score and the differences among the groups were significant (all P＜0.05).Conclusions The serum levels of IL-6,IL-8 and TNF-α increase closely with the volume of infarction and the clinical neurologic impairment degree score,which suggests that IL-6,IL-8 and TNF-α may play an important role in the pathogenesis of ACI,and probably be a prognostic factor of ACI.     

Effect of Xiaofenghuoxue decotion on Th1 and Th2 cytokine production in BALF in allergen-induced asthma guinea pigs

Objective To investigate the effect of Xiaofenghuoxue decotion(XFHX)on the production of Th1 and Th2 cytokine in bronchoalveolar lavage fluid(BALF)in asthma guinea pigs sensitized and challenged with ovalbumin.Methods The animals were randomly divided into 5 groups:the normal control group,the asthma group,the XFHX high dose group,the XFHX low dose group and the budesonide group.2 weeks after sensitization with a mixture of 10% OVA and 10% Al(OH)3,the drugs were given to the animals for 7 days,and then the animals were challenged with aerosol of 1% ovalbumin,the latency time for the onset of respiratory abnormalities was examined,BALF was collected 16 h after.Leukocyte in BALF was counted.The level of IFN-γ reduced by Th1 cells and IL-4 and IL-5 produced by Th2 cells were determined by EILSA.NO contents were also examined.Results The latency time for the onset of respiratory abnormalities and leukocyte infiltration in both XFHX high dose group and low dose group significantly decreased compared with the asthma group.The level of IFN-γ in both XFHX high dose group and low dose group significantly increased compared with the asthma group,the level of IL-4 and IL-5 in both XFHX high dose group and low dose group significantly decreased compared with the asthma group.The production of NO was inhibited in both XFHX high dose group and low dose group compared with the asthma group.Conclusions These results suggest that the anti-asthma effect of Xiaofenghuoxue decotion is associated with NO regulated reversal of Th1/Th2 cytokine balance in allergen-induced asthma guinea pigs.     

Calcineurin/NFAT pathway mediates wear particleinduced TNF-α release and osteoclastogenesis from mice bone marrow macrophages in vitro

Aim： To investigate the roles of the calcineurin/nuclear factor of activated T cells （NFAT） pathway in regulation of wear particles-induced cytokine release and osteoclastogenesis from mouse bone marrow macrophages in vitro. Methods： Osteoclasts were induced from mouse bone marrow macrophages （BMMs） in the presence of 100 ng/mL receptor activator of NF-KB ligand （RANKL）. Acridine orange staining and M＇IF assay were used to detect the cell viability. Osteoclastogenesis was determined using TRAP staining and RT-PCR. Bone pit resorption assay was used to examine osteoclast phenotype. The expression and cellular localization of NFATcl were examined using RT-PCR and immunofluorescent staining. The production of TNFa was analyzed with ELISA. Results： Titanium （Ti） or polymethylmethacrylate （PMMA） particles （0.1 mg/mL） did not significantly change the viability of BMMs, but twice increased the differentiation of BMMs into mature osteoclasts, and markedly increased TNF-α production. The TNF-α level in the PMMA group was significantly higher than in the Ti group （96 h）. The expression of NFATcl was found in BMMs in the presence of the wear particles and RANKL. In bone pit resorption assay, the wear particles significantly increased the resorption area and total number of resorption pits in BMMs-seeded ivory slices. Addition of 11R-VlVIT peptide （a specific inhibitor of calcineurin-mediated NFAT activation, 2.0 pmol/L） did not significantly affect the viability of BMMs, but abolished almost all the wear particle-induced alterations in BMMs. Furthermore, VlVlT reduced TNF-α production much more efficiently in the PMMA group than in the Ti group （96 h）. Conclusion： Calcineurin/NFAT pathway mediates wear particles-induced TNF-α release and osteoclastogenesis from BMMs. Blockade of this signaling pathway with VlVlT may provide a promising therapeutic modality for the treatment of periprosthetic osteolysis.