Objective:To investigate the impact of recombinant flagellin targeting TLR5 and NLRC4 simultaneously or respectively on innate immune cells in mice. Methods: Induction,expression,purification and identification of recombiant FliC,which were FliC(activating both TLR5 and NLRC4);FliCΔ90-97(unable to activate TLR5),FliC-L3A(unable to activate NLRC4),FliCΔ90-97:L3A(unable to activate both TLR5 and NLRC4). The mice were divided into five groups,namely group FliC,FliC-L3A,FliCΔ90-97,FliCΔ90-97:L3A and PBS,which were injected with 100μl PBS or 10μg recombinant flagellin intraperitoneally,three mice in each group. 12 h later,the mice were executed using dislocation of cervical vertebra and the splenic and peritoneal cells were isolated. The spleen was grinded into single-cell suspension. The proportion of neutrophils,NK cells,DCs and the expression level of CD80 and CD86 on DCs were evaluated with flow cytometry. Results:Group FliC,group FliC-L3A and group FliCΔ90-97 shared the similar proportion of neutrophils in peritoneal cavity ( P>0. 05 ) , and all of which were significantly higher than group PBS and group FliCΔ90-97 ( P<0. 01),and NK cells also showed the similar trend. Compared with group FliCΔ90-97 and FliCΔ90-97:L3A,the mean fluorescence intensities(MFIs) of CD80 and CD86 in group FliC and FliC-L3A increased significantly(P<0. 01). The proportion of Treg in spleen was highest among all groups. Conclusion:Activation of TLR5 and NLRC4 had similar chemotaxis of neutrophils and NK cells. The ex-pression of CD80 and CD86 on DCs were upregulated after stimulation by flagellin and TLR5-dependent. Activation of TLR5,but not NLRC4,increased the proportion of Treg in spleen.

Objective:To explore the impact of TLR5 and NLRC4 activation on the proliferation of different breast cancer cell lines,MCF-7 and MDA-MB-23 i.Methods:Induction,expression,purification and identification of recombiant flagellin,including FliC (activating both TLR5 and NLRC4),FliC△90-97 (unable to activate TLR5),FliC-L3A (unable to activate NLRC4),FliC△90-97:L3A (unable to activate both TLR5 and NLRC4).Using different concentration of recombinant flagellin to stimulate MCF-7 and MDA-MB-231 cell lines,72 h later,the proliferation of tumor cells were detected with CCK8.We also used soft AGAR forming experiments to detect the inhibition ratio of recombinant flagellin on breast cancer cell lines.Briefly,1 000 cells were plated in the 6-well plate,then stimulated with 1 μg/ml recombinant flagellin,14 days later,the number of cloning were counted after crystal violet staining.Results:After stimulation with four recombinant flagellins at the concentration of 0.1 μ,g/ml,the inhibition ratio on MCF-7 reached 30％,and FliC△90-97 were dose-dependent on the inhibition of MCF-7 proliferation.At the concentration of 1 μg/ml,FliC-L3A which only activated TLR5 showed stronger inhibition ratio than FliC.FliC△90-97:L3A which did not activate both TLR5 and NLRC4 also inhibited the proliferation of MCF-7.After adding transfection reagent,four recombinant flagellins showed inhibition effect on MDA-MB-231.Conclusion:Flagellin can inhibit the proliferation of MCF-7 and MDA-MB-231,and the mechanism of inhibition on the proliferation were not TLR5 and NLRC4 pathway dependent.There might exist new mechanisms to explain this phenomenon.

Objective:To explore the expression and location of TLR5 and NLRC4 on different breast cancer cell lines MDA-MB-231,MCF-7 and MDA-MB-435 and TLR5 activation in breast cancer cell line by recombinant flagellin . Methods:The mRNA level of TLR5 and NLRC4 in MDA-MB-231, MCF-7 and MDA-MB-435 cell were detected with quantitative Real-time PCR and TLR5 expression and location in MDA-MB-231 and MCF-7 cell were detected with Flow cytometry. Induction,expression,purification and i-dentification of recombiant flagellin,including FliC (activating both TLR5 and NLRC4),FliC△90-97(unable to activate TLR5),FliC-L3A (unable to activate NLRC4),FliC△90-97:L3A (unable to activate both TLR5 and NLRC4). 1 μg/ml recombinant flagellin were used to stimulate MCF-7 cell lines,12 h later,the supernate were collected,and ELISA was performed to assess the secretion of IL-8. Results:The mRNA level of TLR5 in MCF-7 cell was 1 700 folds higher than that of MDA-MB-435. TLR5 was expressed in MCF-7 cell surface and ctyosol,while expressed only in cytosol in MDA-MB-231 cell. FliC and FliC-L3A,which were able to activate TLR5 pathway,stimualted MCF-7 cell line to secret IL-8,but FliC△90-97 and FliC△90-97:L3A did not. Conclusion:TLR5 and NLRC4 have been expressed in different breast cancer lines,but there exists difference on the expression level and location of TLR5. Expression level of TLR5 and NLRC4 in MCF-7 cell were higher than other breast cancer lines. TLR5 receptor which is expressed on the surface of breast cancer cell can be activated by flagellin,and these work also provide us experimental basis to further understand the impact of TLR5 activation on breast cancer cell proliferation.

Purpose To evaluate the correlations between high resolution CT (HRCT) findings and IASLC/ATS/ERS pathological classification of ground glass nodule (GGN). Materials and Methods 121 patients with confirmed GGN were selected, and divided into benign group (22 cases), PIL group (21 cases), microinvasive carcinoma group (26 cases) and invasive carcinoma group (52 cases), then the imaging, pathology and prognosis data of patients with pulmonary GGN were reviewed, and the differences among GGN of different pathological types were analyzed.Results Maximum diameter, margin, vacuole sign, solid component, shape and blood vessels through of GGN were significantly different among the four groups (χ2=9.945-31.068,P<0.05). Maximum diameter and margin were significantly different between invasive adenocarcinoma and other groups (P<0.008); vacuole sign of the benign group was significantly different with other groups (P<0.008); the existence of solid component and shape were significantly different between invasive adenocarcinoma and minimally invasive adenocarcinoma (P<0.008); there was significant difference of blood vessels through between invasive adenocarcinoma and benign lesions (P<0.008). Among the 121 lesions, no metastasis except one invasive adenocarcinoma case complicated with distant metastasis.Conclusion Maximum diameter of GGN greater than 16.35 mm, with spiculation or lobulation represent invasive adenocarcinoma; vacuole sign within the GGN represent malignancy; with solid component and irregular shape can be used to identify invasive adenocarcinoma from minimally invasive adenocarcinoma; while blood vessels through can be used to identify invasive adenocarcinoma from benign lesions; the prognosis of GGNs is well with only 0.83% probability of distant metastasis.

OBJECTIVETo investigate the effects of ferulic acid on the migration of vascular smooth muscle cells (VSMCs) and its correlated mechanisms.

METHODSVSMCs were in vitro cultured. Under the vascular endothelial growth factor (VEGF) induced conditions, the VSMCs migration were detected using the scratch test and the invasion assay after intervened by ferulic acid. The effects on the mRNA expressions of matrix metalloproteinase, (MMP)-2 and MMP-9 were detected using RT-PCR. The effects on the protein expressions of the tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 were detected using Western blot.

RESULTS(1) Ferulic acid (10(2) ng/mL and 10(3) ng/mL) could inhibit VEGF-induced VSMCs migration. (2) Ferulic acid (10(2) ng/mL and 10(3) ng/mL) could down-regulate the VEGF-induced VSMCs migration by inhibiting MMP-9 mRNA expression. (3) Ferulic acid (10(2) ng/mL and 10(3) ng/mL) up-regulated VEGF-induced VSMCs TIMP-2 protein expressions.

CONCLUSIONFerulic acid (10(2) ng/mL and 10(3) ng/mL) could inhibit VEGF-induced VSMCs migration by inhibiting the MMP-9 mRNA expression, and increasing the TIMP-2 protein expression.

Cell Movement ; drug effects ; Cells, Cultured ; Coumaric Acids ; pharmacology ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo investigate the protective effects of sildenafil to noise-induced hearing loss in guinea pigs.

METHODSGuinea pigs were randomly divided into control group, noise exposure group and the sildenafil treatment group, with 10 in each group. One week after the exposure of 110 dB (A) white noise, sildenafil [10 mg/(kg×d)] and normal saline [4 ml/(kg×d)] were injected into guinea pigs of noise plus sildenafil group (NSG) and noise plus normal saline group(NNG) respectively. One week after noise exposure to four weeks continuous administration. ABR thresholds were measured respectively prior to the experiment, 1 week post-noise, 1, 2 and 4 weeks post-drugs. The changes of cochlea hair cells were also observed by scanning electron microscope (SEM).

RESULTSAfter noise exposure, the ABR threshold shifts in NSG were significantly fewer than that in the NNG. An average of 19.1 dB in NNG compared with 19.8 dB in NSG. Four weeks after exposure, the threshold shifts were become larger to 22.0 dB in NNG while become smaller to 4.8 dB in NSG. Compared NNG with NSG, in addition to noise exposure time point, there were statistically significant differences in the rest time points after administration of the ABR threshold (P<0.05). SEM showed that the inner and outer hair cells in NNG displayed mess, fusion and imperfections. In NSG, the hair cells displayed slight pathological changes, there was no significant difference when compared with control group.

CONCLUSIONSildenafil is able to reduce the ascending of ABR thresholds shift, and it can significantly protect against noise-induced hearing loss.

Animals ; Auditory Threshold ; drug effects ; Cochlea ; drug effects ; Guinea Pigs ; Hair Cells, Auditory ; drug effects ; Hearing Loss, Noise-Induced ; physiopathology ; Piperazines ; pharmacology ; Purines ; pharmacology ; Sildenafil Citrate ; Sulfones ; pharmacology

Objective To summarize our experience of simple neuroendoscopic endonasal transsphenoidal surgery for pituitary adenoma and discuss its advantages. Methods Forty-two patients with pituitary adenoma diagnosed by CT or MRI detection (diameters of the tumor: <1cm in 11 patients, 1-2cm in 11, 2-3cm in 18, >3cm in 8) were treated by simple neuroendoscopic endonasal transsphenoidal surgery. During transsphenoidal approach, the nasal mucosa was not incised and the nasal septum and median nasal conchae were not dissected. Under neuroendoscopic assistance, the anterior wall of the sphenoidal sinus and sellae base was opened directly and adenoma resection was performed. Results All the patients were followed up for 1-12 months. By postoperative MRI detection, adenomas were resected completely in 38 patients, subtotally in 4 cases. Rapid improvement of vision was noted postoperatively. The increased hormonal level in 18 of 22 patients was decreased to normal postoperatively and another 4 cases improved clinically in serum hormone levels during follow-up. Postoperative transient CSF leakage occurred in 6 cases and transient diabetes insipidus in 23 cases. Conclusions The simple neuroendoscopic endonasal transsphenoidal surgery for pituitary adanoma is safe, time-saving and without severe complications, provides a satisfactory deep illumination and a panoramic operative field. The injury to the patient and postoperative complications are less. The clinical application of the neuroendoscopy for pituitary adenoma is worthy of recommending.

OBJECTIVETo compare the conventional pulmonary function test results of children with asthma or cough variant asthma (CVA).

METHODSA total of 140 children, who were diagnosed with asthma or CVA from May 2010 to May 2011, were divided into acute asthma attack (n=50), asthma remission (n=50) and CVA groups (n=40); 30 healthy children were included as a control group. The forced vital capacity (FVC), forced expiratory volume in one second (FEV1), peak expiratory flow (PEF), forced expiratory flow after 25% of vital capacity has been expelled (FEF25), forced expiratory flow after 50% of vital capacity has been expelled (FEF50), forced expiratory flow after 75% of vital capacity has been expelled (FEF75) and maximal midexpiratory flow (MMEF75/25) were measured.

RESULTSThe mean percent predicted values of all the above indices were lower than 80% in the acute asthma attack group, with FEF50, FEF75 and MMEF75/25 declining markedly; the mean percent predicted values of FEF75 and MMEF75/25 were lower than 80% in the CVA group. All the pulmonary function indices in the acute asthma attack group were lower than those in the control group. The mean percent predicted values of FVC, FEV1, FEF25 and MMEF75/25 in the asthma remission and CVA groups were lower than in the control group. All the pulmonary function indices in the acute asthma attack group were lower than in the asthma remission and CVA groups, but there were no significant differences between the asthma remission and CVA groups.

CONCLUSIONSThere is small and large airway dysfunction, particularly small airway dysfunction, in children with acute asthma attack. Children with CVA present mainly with mild small airway dysfunction, as do those with asthma in remission.

Asthma ; physiopathology ; Child ; Cough ; physiopathology ; Female ; Forced Expiratory Volume ; Humans ; Lung ; physiopathology ; Male ; Vital Capacity

Along with the extensive application of intervention in treating coronary heart disease (CHD), problems such as post-interventional restenosis, no reflowing, and slow flowing, and so on increasingly emerge. Chinese medicine and pharmacy occupy advantages covering preventing post-intervention risk factors, improving and relieving patients' symptoms, and so on. It is a good support and supplement for routine therapies in modern medicine. Taking the pathological stages of CHD and clinical problems of coronary intervention as the breakthrough points, the Chinese medical syndromes and correspondent Chinese medicine recipes and herbs were preliminarily studied by combining Chinese medicine's recognition and clinical observation on coronary intervention.
Angioplasty, Balloon, Coronary ; Coronary Disease ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Phytotherapy ; methods

OBJECTIVETo study the effect of Xianlong granules (XLG) on immunological function in the rat of adjuvant arthritis (AA).

METHODRats were randomly divided into normal group, AA model group, prednisone group and low, middle and high dose XLG groups, 10 rats in each group. All rats were treated by intragastric administration from the 18 days after arthritis was induced by the complete Freud's adjuvant and the effect of XLG on toes swelling was observed. On the 30th days after modeling, proliferation of the splenic and thymic lymphocytes, and IgG secreted by splenocytes were detected respectively by MTT assay and ELISA.

RESULTCompared with the model group, both the high and middle dose XLG groups had significant therapeutic effects on toes dwelling in the rat of AA (P < 0.05 or P < 0.01); The low, middle and high dose XLG groups strengthened the PHAM-inhibited proliferation of splenic lymphocytes (P < 0.05), and inhibited the PHAM-augmented proliferation of thymic lymphocytes (P < 0.05); XLG did not significantly effect on IgG level secreted by splenocytes in rats of AA.

CONCLUSIONXLG can cure toes swelling in rats of AA, which is related with regulation of the abnormal immunlological function.

Animals ; Arthritis, Experimental ; immunology ; pathology ; prevention & control ; Cell Proliferation ; drug effects ; Colubridae ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Edema ; immunology ; pathology ; prevention & control ; Female ; Immunoglobulin G ; metabolism ; Lymphocytes ; drug effects ; pathology ; secretion ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Spleen ; pathology ; secretion ; Thymus Gland ; pathology ; Toes ; pathology