Long-term in vivo imaging of dendritic spines in the hippocampus reveals structural plasticity.

Hippocampal function is important for learning and memory. During memory processing, hippocampal CA1 neurons play a crucial role by integrating excitatory synaptic input from CA3 and the entorhinal cortex. These neurons receive excitatory input almost exclusively on dendritic spines. The formation and elimination--structural plasticity--of dendritic spines reflect wiring changes within the hippocampal network. Despite the relevance of the hippocampus in learning and memory, most in vivo data on structural plasticity derive from cortical regions. We established a chronic hippocampal window approach using two-photon microscopy to visualize dendritic spines throughout all CA1 hippocampal layers and over a time course of weeks. Moreover, even granule cells in dentate gyrus could be reliably detected. We found that the spine density in stratum radiatum (∼1.1 per micrometer) remained stable over weeks. However, a small fraction (3.4%) of spines were formed and eliminated between imaging sessions, which demonstrated that spines of CA1 neurons exhibit structural plasticity in adult mice. In addition, we tested for possible inflammatory or behavioral side effects of hippocampal window implantation. Mice exhibited a transient increase in microgliosis and astrogliosis, which declined within a few weeks. We did not detect any difference in behavioral performance in an open-field and contextual fear-conditioning paradigm. In conclusion, hippocampal long-term two-photon imaging revealed structural plasticity of dendritic spines in CA1 pyramidal neurons. This approach may provide a powerful tool to analyze changes in neuronal network rewiring during hippocampal learning and memory processes in health and disease.

[A research on the influence of two herbal concoctions on Toll-like receptor signal pathways of influenza virus induced pneumonia in mice].

OBJECTIVE: To investigate the effect of Shufeng Xuanfei and Jiebiao Qingli concoctions on Toll-like receptor (TLR) signal pathway of pneumonia infected with influenza virus in mice. METHODS: The pneumonia model was reproduced by nasal dropping of influenza virus A in mice. The mice were randomly divided into nine groups: normal group (C), model group (M), tamiflu group (D), Shufeng Xuanfei low-dose (SL), medium-dose (SM) and high-dose (SH) groups, Jiebiao Qingli low-dose (JL), medium-dose (JM) and high-dose (JH) groups, each n=12. Two hours after model-reproduction, the mice in C group and M group received distilled water by gavage. The mice in D group received 2.5 g×mL(-1)×d(-1) oseltamivir phosphate. Shufeng Xuanfei formula in doses of 3.76, 1.88, 0.94 g×kg(-1)×d(-1) were respectively administered to SH, SM and SL groups by gavage, Jiebiao Qingli formula in doses of 4.37, 2.18, 1.09 g×kg(-1)×d(-1) was given to JH, JM and JL groups by gavage, respectively. Each group was in equal dose of 0.2 mL daily over a 4-day period. Total RNA was extracted in each group. Then gene chips were used to screen these RNA samples. Some genes that were involved in TLR signal pathways were selected. These candidate genes were verified by real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: TLR7, MYD88, CCL5, IFNB1, IL6, IL12a, NFKBIA and IKBKB were up-regulated in model group compared with control group. Compared with model group, down-regulated genes in medium-dose, low-dose Shufeng Xuanfei formula and medium-dose Jiebiao Qingli formula included TLR3, TLR7, MYD88, CCL5, IFNB1, IL6, IL12a, NFKBIA and IKBKB (log2 signal intensity of SM/M in medium-dose Shufeng Xuanfei formula group were -1.24, -2.02, -1.36, -1.95, -0.63, -1.33, -3.50, -1.33, -1.33, log2 signal intensity of SL/M in low-dose Shufeng Xuanfei group were -1.07, -2.43, -2.63, -2.30, -5.09, -3.19, -3.53, -1.95, -1.95, log2 signal intensity of JM/M in medium-dose Jiebiao Qingli formula group were -1.78, -0.55, -1.35, -1.47, -1.65, -2.03, -3.02, -1.57, -1.57, respectively). The results suggested that the effect of Shufeng Xuanfei formula was better than that of Jiebiao Qingli formula. By RT-PCR, compared with model group, low-dose, medium-dose and high-dose groups of Shufeng Xuanfei formula, medium-dose and high-dose groups of Jiebiao Qingli formula, and tamiflu group, significant decrease in TLR7, nuclear factor-ΚB (NF-ΚB), myeloid differential protein-88 (MyD88) mRNA expression were found. Medium-dose and low-dose Shufeng Xuanfei formula group (TLR7 mRNA: 3.6±0.3, 3.5±1.2 vs. 7.4±1.6, NF-ΚB mRNA: 1.1±0.2, 1.0±0.2 vs. 2.2±0.4; MyD88 mRNA: 1.4±0.4, 1.0±0.3 vs. 3.4±0.9, all P<0.01) and medium-dose Jiebiao Qingli formula group (TLR7 mRNA: 4.9±0.3 vs. 7.4±1.6, NF-ΚB mRNA: 1.3±0.7 vs. 2.2±0.4, MyD88 mRNA: 1.6±0.8 vs. 3.4±0.9, P<0.05 or P<0.01) were shown statistically significant decreases compared with the model group. CONCLUSIONS: Medium-dose and low-dose Shufeng Xuanfei formula and medium-dose Jiebiao Qingli formula can inhibit the inflammatory reaction induced by influenza virus by down-regulating the NF-ΚB through TLR signal pathways dependent on MyD88. The regulation of Shufeng Xuanfei formula in TLR signal pathways was superior to that of Jiebiao Qingli formula.

Baicalin inhibits TLR7/MYD88 signaling pathway activation to suppress lung inflammation in mice infected with influenza A virus.

The present study aimed to investigate the protective effects and underlying mechanisms of baicalin on imprinting control region mice infected with influenza A/FM/1/47 (H1N1) virus. Oral administration of baicalin into mice infected with H1N1 prevented death, increased the mean time to death and inhibited lung index and lung consolidation. Subsequently, fluorescence quantitative polymerase chain reaction was used to assess the mRNA expression of toll-like receptor 7 (TLR7) and myeloid differentiation primary response gene 88 (MYD88), and western blot analysis was used to determine the expression of phosphorylated nuclear factor κB (NF-κB)-P65 and c-jun/activator protein 1 (AP-1). An enzyme-linked immunosorbent assay was applied to test for the inflammatory cytokines, tumor necrosis factor (TNF)-α and interleukin (IL)-1ß and IL-6, in the lung tissue. The findings indicated that baicalin downregulated the mRNA expression of TLR7 and MYD88, significantly downregulated the protein expression of NF-κB-P65 and AP-1 and also inhibited the secretion of TNF-α, IL-1ß and IL-6. In conclusion, baicalin effectively reduced the pathological damage and inflammation of the lungs by downregulating the TLR7/MYD88-mediated signaling pathway.

[The expression of the inflammation-related cytokines in pneumonia mice infected with influenza virus and regulation of Shufengxuanfei and Jiebiaoqingli herbal anti-virus formulas].

OBJECTIVE: To investigate the expression of the inflammation-related cytokines in pneumonia mice infected with influenza virus and regulation of Shufengxuanfei(SFXF) and Jiebiaoqingli (JBQL) Chinese herbal anti-virus formulas. METHODS: Mice were anesthetized and then infected intranasally by dropping 0.05 mL of influenza virus suspension (4×LD50;) except normal group. Mice were divided randomly into nine groups: normal group, model group (virus only), control group [11.375 g/(kg.d)Oseltamivir], low-dose SFXF [0.94 g/(kg.d)], medium-dose SFXF [1.88 g/(kg.d)], high-dose SFXF [3.76 g/(kg.d)], low-dose JBQL [1.09 g/(kg.d)], medium-dose JBQL [2.18 g/(kg.d)] and high-dose JBQL [4.36 g/(kg.d)]. Oseltamivir group, SFXF groups and JBQL groups were administered to mice by oral gavage in equal dose of 0.2 mL daily for 4 consecutive days, while the rest of the groups received water only. Total RNA was extracted in each group. Then gene chips were used to screen these RNA samples. Select differentially expressed genes of cytokines involved in inflammation. Some candidate genes, such as IL-1ß, IL-8, IL-10, RANTES and ICAM-1 were verified by qRT-PCR. To confirm the genes expression data from the microarray involved in inflammation in response to virus infection and treatment, we used qPCR to verify mRNA relative expressions of IL-1ß, IL-8, IL-10, RANTES and ICAM-1. The expression of IL-1ß protein in lung tissues was verified by Western blotting. RESULTS: IL-1ß, CXCR2, CCL5, IL-10, IL-6, IL-18, TGF-ß1 and CCL2 were up-regulated in model group. Gene expressions of IL-1ß, CXCR2, CCL5, IL-10 and IL-6 were significantly down-regulated by all therapeutic groups. SFXF in medium-dose and low-dose down-regulated gene expressions of IL-18, TGF-ß1, CCL2 and CCL5. IL-18 and CCL5 was down-regulated by both low-dose and medium-dose JBQL. qRT-PCR and western blot experiments showed that two formulas in medium-dose can down-regulate mRNA and protein expression of IL-1ß (P<0.01). Both SFXF and JBQL in medium-dose significantly decreased the IL-8, RANTES, ICAM-1 and IL-10 mRNA expression (P<0.05 or P<0.01), compared with the model group. As expected, qRT-PCR data were in good agreement with the microarray assay. CONCLUSION: The two anti-viral formulas may inhibit inflammatory immunopathogenesis, and may have the actions of protection the lung tissue from influenza-induced injury.

Gene Expression Profiles Underlying Selective T-Cell-Mediated Immunity Activity of a Chinese Medicine Granule on Mice Infected with Influenza Virus H1N1.

A Chinese medicine granule, Shu-Feng-Xuan-Fei (SFXF), is critical for viral clearance in early phase of influenza virus infection. In this study, 72 ICR mice were randomly divided into six groups: normal control group, virus control group, Oseltamivir group, low-dose SFXF, medium-dose SFXF, and high-dose SFXF. Mice were anesthetized and inoculated with 4LD50 of influenza virus A (H1N1) except normal control group. Oseltamivir group received 11.375 mg·kg(-1) ·d(-1) Oseltamivir Phosphate. SFXF 3.76, 1.88 and 0.94 g·kg(-1) ·d(-1) were administrated to mice in all SFXF groups. Each group was in equal dose of 0.2ml daily for 4 consecutive days. Mice were sacrificed and then total RNA was extracted in lung tissue. Some genes involved in T-cell-mediated immunity were selected by DNA microarray. These candidate genes were verified by Real-Time PCR and western immunoblotting. Compared with virus control group, in Toll-like receptor signaling pathway, 12 virus-altered genes were significantly reduced following medium-dose SFXF treatment. Eighteen antigen processing presentation-associated genes were upregulated by medium-dose SFXF. In the process of T cell receptor signaling pathway, 19 genes were downregulated by medium-dose SFXF treatment. On exploration into effector T cells activation and cytokines, all of altered genes in virus control group were reversed by medium-dose SFXF. Real-time PCR and western immunoblotting showed that the regulation of medium-dose SFXF in IL-4, IFN-γ, TNF-α, IL-1ß, TLR7, MyD88, p38, and JNK was superior to Oseltamivir and high-dose SFXF group. Therefore, SFXF granules could reduce influenza infected cells and activation of T cells.

[Effect of shufeng xuanfei recipe and jiebiao qingli recipe on mRNA and protein expressions of TLR7, MyD88, and NF-kappaB in mice infected with influenza virus].

OBJECTIVE: To observe effect of Shufeng Xuanfei Recipe (SXR) and Jiebiao Qingli Recipe (JQR) on mRNA and protein expressions of Toll-like receptor 7 (TLR7), myeloid differentiation factor 88 (MyD88), and nuclear factor-kappaB (NF-kappaB) in mice infected with influenza virus FM1. METHODS: One hundred and eight mice were randomly divided into nine groups, i.e., the normal control group, the model group, the Oseltamivir group (at the daily dose of 2.5 g/mL), the high dose SXR group (at the daily dose of 3.762 g/kg), the middle dose SXR group (at the daily dose of 1.881 g/kg), the low dose SXR group (at the daily dose of 0.941 g/kg), the high dose JQR group (at the daily dose of 4.368 g/kg), the middle dose JQR group (at the daily dose of 2.184 g/kg), and the low dose JQR group (at the daily dose of 1.092 g/kg), 12 in each group. All mice were mildly anesthetized by ether. Mice in the normal control group were treated by nasal drop of 0.05 mL normal saline, while mice in the rest groups were infected by nasal drop of 0.05 mL influenza virus strain FM1 (LD50). The successful modeling rate was 100%. All medication was performed by gastrogavage 2 h after infection. Distilled water was given by gastrogavage to mice in the normal control group and the model group at the daily dose of 0.2 mL, each time per day for 4 successive days. mRNA expressions of TLR7, MyD88, and NF-kappaB in the lung tissue were determined by Western blot. RESULTS: Compared with the normal control group, mRNA expressions of TLR7, MyD88, and NF-kappaB increased in the model group (P < 0.01). Compared with the model group, mRNA and protein expressions of TLR7, MyD88, and NF-kappaB decreased in the Oseltamivir group, the high, middle, and low dose SXR groups (P < 0.05, P < 0.01); mRNA and protein expressions of TLR7 and NF-kappaB decreased in the high and middle dose JQR groups (P < 0.05, P < 0.01); mRNA expressions of MyD88 decreased in the high and middle dose JQR groups (P < 0.05); protein expressions of MyD88 decreased in the middle dose JQR group (P < 0.05); protein expressions of TLR7 and NF-kappaB decreased in the low dose JQR group (P < 0.05). Compared with the Oseltamivir group, protein expressions of MyD88 decreased in the low dose SXR group (P < 0.05); protein expressions of NF-kappaB decreased in the middle and low dose SXR groups (P < 0.01); mRNA and protein expressions of TLR7 (P < 0.05, P < 0.01), and protein expressions of MyD88 (P < 0.01) decreased in the high, middle, and low dose JQR groups; mRNA and protein expressions of NF-kappaB decreased in the low dose JQR group (P < 0.05, P < 0.01). CONCLUSIONS: Each dose SXR and middle dose JQR could down-regulating the activity of NF-kappaB through adjusting MyD88 dependent TLR signal pathway, thus fighting against influenza virus. SXR was more effective than JQR.

[Screening for genes associated with natural killer cell mediated cytotoxicity in pneumonia mice infected with influenza virus and regulation of two herbal anti-virus formulas].

OBJECTIVE: To investigate the regulation of two herbal anti-virus formulas on gene expression profile associated with natural killer cell (NK cell) mediated cytotoxicity in pneumonia mice infected with influenza virus. METHODS: According to random number table, 90 ICR mice were divided into nine groups with 10 mice in each group: normal group (N), model group (M), oseltamivir group (control group, C), low-dose, medium-dose and high-dose Shufeng Xuanfei formula groups (SL, SM, SH groups), and low-dose, medium-dose and high-dose Jiebiao Qingli formula groups (JL, JM, JH groups). The model of pneumonia was reproduced by nasal dropping influenza virus A (FM1) in mice. N group was given isotonic saline 0.05 ml in nasal drops. After 2 hours of model-building, C group was received 11.375 mg×kg⁻¹×d⁻¹ oseltamivir phosphate. Shufeng Xuanfei formula (mainly honeysuckle, forsythia and radix isatidis, etc.) with 3.76, 1.88 and 0.94 g×kg⁻¹×d were administrated to SH, SM and SL groups by gastric irrigation respectively. Jiebiao Qingli formula (mainly ephedra, gypsum, glycyrrhiza glabra, etc.) with 4.36, 2.18 and 1.09 g×kg⁻¹×d⁻¹ were administrated to JH, JM and JL groups by gastric irrigation respectively. In N and M groups, normal saline was administrated with gastric perfusion. Each group was in equal dose of 0.2 ml daily over a 4-day period. Total RNA in lung tissue of mice were extracted in each group, then gene chips were used to screen these RNA samples. Some genes involved NK cell mediated cytotoxicity were selected, with "I" representing of signal intensity. These candidate genes were verified by real-time fluorescent quantitation polymerase chain reaction (PCR) and Western blotting. RESULTS: In the pathway of NK cell mediated cytotoxicity, M group up-regulated 43 genes expression, and 36, 29, 22, 21, 20 and 10 genes showed down-regulation in SM, JM, SL, JH, SH and JL groups, respectively. Apart from gene co-expression network in SH, SL, JH, JM and JL, SM also expressed other differential genes which SH, SL, JH, JM and JL did not. So medium-does Shufeng Xuanfei formula had the most significant regulation in gene expression of NK cell mediated cytotoxicity. By real-time PCR and Western blotting experiments showed that compared with the M group, mRNA and protein expression of tumor necrosis factor-α (TNF-α) in these two formula groups were significantly down-regulated, especially prominent in SM group and JM group (TNF-α mRNA: 1.07 ± 0.19, 1.19 ± 0.14 vs. 3.20 ± 0.56, both P<0.01). CONCLUSIONS: Influenza viral replication in host cell, which means influenza antigens exposure in infected cells as target cells. NK cells recognize and exert cell mediated cytotoxic function against influenza antigens. Genes associated with NK cell mediated cytotoxicity in influenza infection were up-regulated. Shufeng Xuanfei and Jiebiao Qingli formulas could down-regulate these genes. The mechanism of down-regulated genes is that the number of influenza infected cells and NK cells activation decreases in treatment with two formulas.

Effect and mechanisms of Gong-tone music on the immunological function in rats with Liver (Gan)-qi depression and Spleen (Pi)-qi deficiency syndrome in rats.

OBJECTIVE: To investigate the effects and mechanisms of Gong-tone music on the immunological function in rats with the Chinese medicine syndrome of Liver (Gan)-qi stagnation and Spleen (Pi)-qi deficiency (LSSD). METHODS: Twenty five male Wistar rats of SPF grade were randomly divided into 5 groups: normal group, model group, Xiaoyao Powder () group, Gong-tone group and combined group (the combination of Gong-tone and Xiaoyao Powder), with 5 rats in each group. The rat model for the Chinese medicine syndrome of LSSD was induced by chronic bandage and irregular diet. The course of treatment was 21 days. After the treatment, the levels of serum gastrin and IgG were detected by enzyme-linked immunoabsorbent assay (ELISA). Phagocytosis of macrophages was detected by the neutral red uptake assay and T cell proliferation was investigated by 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: The serum gastrin, macrophage phagocytosis, IgG level and proliferation ability of T cells in the model group were significantly decreased compared with those in the normal group (P <0.05). Compared with those in the model group, the serum levels of gastrin, macrophage phagocytosis, IgG level and proliferation ability of T cells in Gong-tone, Xiaoyao Powder, and combined groups were significantly increased (P <0.05). The combined group was superior to either Gong-tone group or Xiaoyao Powder group. CONCLUSION: Gong-tone music may upregulate the immunological function and play a role in adjuvant therapy in the Chinese syndrome of LSSD.

Effects of quercetin on CDK4 mRNA and protein expression in A549 cells infected by H1N1.

This study was conducted to investigate the effects of quercetin on the expression of cyclin-dependent kinase (CDK4) mRNA and protein in A549 lung epithelial tumor cells infected by H1N1. First, the Thiazolyl Blue Tetrazolium Bromide (MTT) method was used to determine H1N1 virulence, quercetin cytotoxicity and inhibition of the cytopathic effect of H1N1 on A549 cells by quercetin. Subsequently, 100 TCID50 H1N1 was used to infect A549 cells for 2 h prior to culture in maintenance media containing 10 mg/l quercetin. After 4, 12, 24 and 48 h of culture, the cells were collected and total RNA and protein were extracted. Fluorescent quantitative polymerase chain reaction and western blot analysis were then performed to assess the expression of CDK4 mRNA and protein. The experiment demonstrated that the TCID50 of H1N1 in A549 cells was 10-4.75, the maximum non-toxic concentration of quercetin in A549 cells was 30-60 mg/l and the minimum effective concentration of quercetin for the inhibition of the H1N1 cytopathic effect on A549 cells was 10 mg/l. The results indicated that quercetin may significantly inhibit CDK4 mRNA and protein overexpression caused by H1N1 within 4-48 h. In conclusion, quercetin may protect against H1N1 infection by effectively reducing the mRNA and protein expression of CDK4 caused by H1N1 infection.

[Antagonism of baicalin on cell cyclical distribution and cell apoptosis in A549 cells infected with influenza A (H1N1) virus].

To observe the inhibitive effect of Baicalin against influenza A H1N1 virus infection in epithelial cell line A549, the cell proliferation and cytotoxicity were assayed by MTT, the cell cycle and the apoptosis were analyzed by flowcytometer using PI staining, the morphology of cellular nucleolus was observed by Hoechst 33258 staining and the effects of activation on caspase 3 and caspase 8/9 were also detected by immunofluorescent staining with a fluorescence microscope. The results showed that Baicalin exerted an inhibitive effect on CPE after influenza A H1N1 virus infection. The FACS with PI staining showed that the cell cycle of the infected cell was arrested at S phase, the Baicalin-treated group decreased S phase cell ratio and subG0 phase peak in comparison with the control (P < 0.05) and significantly promoted cell proliferation (# P < 0.05). Hoechst33258 staining suggested that Baicalin protected the cellular nucleolus against the influenza virus-induced apoptosis. Observation under the immunofluorescent microscope suggested that the activities of caspase-8 and caspase-3 were enhanced at 36 h post the influenza virus infection, but 100 microg/mL Baicalin suppressing the activation of caspase-8 and caspase-3 rather than that of caspase-9. In summary, this research confirmed that Baicalin inhibited the influenza A H1N1 virus strain infection in vitro, the drug obviously protected cells from apoptosis damages through regulating cell cycle and suppressed the activation of caspase-8 and caspase-3. The down-regulation was significant and showed a dose-dependent relationship.

[Effects of Dureping Injection on the contents of MMP-9 and TIMP-1 in the lung tissue of mice with pneumonia of influenza virus infection].

OBJECTIVE: To observe the effects of Dureping Injection on the contents of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in the lung tissue of mice with pneumonia of influenza virus infection. METHODS: Sixty-six ICR mice were randomly divided into the normal group, the model group, the low, middle, and high dose Dureping Injection groups (0.435, 0.870, and 1.740 mg/d, respectively), and the positive control group (Ribavirin, 2.500 mg/d), 11 in each. The pneumonia of mice with influenza virus infection model was established using influenza virus strain FM1. Mice were intraperitoneally injected with 0. 3 mL FM1 starting from the infection day, once daily. Five days later mice were killed to calculate the lung index. The pathomorphological changes of the lung tissue were observed using routine HE stained sections. The contents of MMP-9 and TIMP-1 in the homogenate of the lung tissue were detected by ELISA double antibody sandwich method. RESULTS: Compared with the normal group, obvious inflammation occurred in the lung tissue of mice in the model group. The lung index, the content of MMP-9, and the value of MMP-9/TIMP-1 increased significantly in the model group (P < 0.01) , while the content of TIMP-1 was not significantly different (P > 0.05). Compared with the model group, the content of MMP-9 in the low and middle dose Dureping Injection groups, and the positive control group was significantly lowered (P < 0.01). The content of TIMP-1 in the low, middle, and high dose Dureping Injection groups, as well as the positive control group significantly increased (P < 0.01) and the value of MMP-9/TIMP-1 decreased (P < 0.01). CONCLUSION: Dureping Injection could alleviate the inflammatory injury of the lung tissue through decreasing the content of MMP-9, elevating the content of TIMP-1 in the lung tissue, and regulating the value of MMP-9/TIMP-1 of mice with pneumonia of influenza virus infection, thus alleviating the inflammatory injury of the lung tissue.

[Effect of dureping injection on T-cells function and their function in killing FM1 infected Mphi in vitro].

OBJECTIVE: To investigate the effect of Dureping Injection (DRP) on the T-cells function of mice and the function of T-cells in killing MF infected by influenza virus subtype A mice-lung adaptive strain FM1 in vitro. METHODS: Number of splenic normal and FM1 infected T-cells in mice were measured by MTT and double-antibody sandwich ELISA, after being treated with DRP at different concentrations (2.1, 8.5 and 17.0 microg/mL), and the effect of DRP on interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) production as well as on splenic T-cell killing FM1 infected Mphi/Ana-1 function were detected. RESULTS: DRP inhibited the multiplication of normal spleen T cells induced by concanavalin A in vitro, suppressed Th2 cell factor IL-10 production, and maintained Th1 cell factor IFN-y at a definite level, moreover, it directly enhanced the power of T-cells in killing FM1 infected Mphi (P < 0.05, P < 0.01). CONCLUSION: DRP could act on mice T-cells to enhance the immune response for antiinfluenza viral FM1 in vitro.

[Effect of Dureping injection on TIR signal pathway on Ana-1 cells].

OBJECTIVE: To investigate the influence of Dureping injection to the murinal celiac macrophage Ana-1 on TIR signal pathway. METHOD: Ana-1 cell line was infected by influenza virus FM1 strain and treated with the Dureping injection in different concentrations (10.1 mg x L(-1) group) for 12 h and 24 h. Then we collected the cells, extracted mRNA and measured the expressions of TLR7, MyD88, IRAK4, TRAF6 and NF-kappaB p65 respectively by RT-PCR. RESULT: Dureping injection down-regulated the expression of TLR7, MyD88, IRAK4, TRAF6 and NF-kappaB p65 mRNA in Ana-1 cell line infected by influenza virus, in a dose-dependent manner significantly. CONCLUSION: Dureping injection has an obvious effect against influenza virus FM1 strain by regulating the TIR signal pathway.

[Experimental study on anti-arthritis effect of jiawei mufangji decoction in rats].

OBJECTIVE: To explore the therapeutic effect of Jiawu Mufangji Decoction (JMD) in treating rats with adjuvant arthritis (AA) and its mechanism. METHODS: AA model rats induced by Freund's complete adjuvant were treated with JMD by gastrogavage starting from 18 days after modeling. On the 39th day, body weight, spleen and thymus index, and swelling degree of paw of the AA rats were measured, pathological changes of the ankle joint tissue were observed using HE staining, and serum levels of interleukin-1beta (IL-1beta) and tumor necrosis factor a (TNF-alpha) were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: JMD could relieve the symptoms of AA rats, decrease the paw swelling, improve the weight and spleen and thymus index, reduce the dropsy of joints and lymphocytes infiltration, inhibit the proliferation of synovium, and obviously lower the serum levels of interleukin-1beta and TNF-alpha. CONCLUSION: The therapeutic effect of JMD might be related to its action in down-regulating the serum levels of IL-1beta and tumor necrosis factor alpha.

[The study on the relationship between the tumor-like proliferation of synoviocytes and the expression of c-myc and ODC genes in adjuvant arthritis rats].

AIM: To investigate the mechanism of the tumor-like proliferation of synoviocytes in adjuvant arthritis (AA) rats. METHODS: Wistar rats were randomly divided into 2 groups, AA group and control group. Freund's complete adjuvant was injected into AA group rats to induce adjuvant arthritis. The knee synovial tissues of the rats were taken and synoviocytes were separated and cultured in vitro. The proliferation of cultured synoviocytes was measured by MTT colorimetry. Meanwhile, the mRNAs of C-myc and ODC genes in synovial tissues of the rats were detected by semi-quantitative RT-PCR. RESULTS: (1)Synoviocytes of AA rats proliferated more markedly than those of control rats (P<0.01), and exhibited a tumor-like proliferation. (2)The mRNAs of C-myc and ODC genes were obviously higher in synovial tissues of AA rats than those of control rats. CONCLUSION: The tumor-like proliferation of synoviocytes from AA rat joint might be related to the increased mRNAs of C-myc and ODC genes.

[Influence of bondage stress on Th1/Th2 cytokines and growth of tumors in mice bearing S180 tumor cells].

AIM: To observe the influence of bondage stress on Th1/Th2 cytokines and growth of tumor in mice bearing S180 tumor cells. METHODS: 2 x 10(6) S180 tumor cells were injected subcutaneously into the murine right oxter,and bondage stress was imposed on mice for 8 hours per day,ten days running. Then T lymphocyte proliferation was detected by MTT colorimetry. IFN-gamma and IL-2 secreted by splenocytes was measured by mitogen-activating lymphoblast assay and macrophage NO production assay. At the same time, the levels of IL-4 and IL-10 in serum were detected by ELISA, and the weight of the tumors and thymuses were measured. RESULTS: Compared with mice only injected with S180 tumor cells, tumor bearing mice suffered from bondage stress showed decreased T lymphocyte proliferation, thymus index, and levels of IFN-gamma and IL-2, but increased levels of IL-4 and IL-10(P<0.05), and tumor weight (P<0.01). CONCLUSION: Bondage stress could aggravate the suppression of cellular immunity in mice bearing S180 tumor and promote the T cells to shift towards Th2 type cells, which may be one of the mechanisms of promoting tumor growth by bondage stress.