This study was aimed to observe the effects ofFeng-Shi-Ning (FSN) capsule on the balance of IL-17+CD4+T (Th17) / Foxp3+CD4+CD25+regulatory T (Treg) cells in peripheral blood and the inflammatory media in synovial fluid of rats with rheumatoid arthritis (RA), in order to further study the mechanism of FSN capsule in RA treatment. The collagen-induced arthritisⅡ (CIA) was used in the RA rat model establishment. The balance of Th17 and Treg cells in peripheral blood was analyzed by the flow cytometry method for effects of FSN capsule treatment. The levels of TNF-α, IL-6, IL-17A, IL-4 and IL-10 in synovial fluid were measured by cytometric bead array (CBA) method. The effects of the normal rats, CIA model rats and the rats treated by tripterygium wilfordii polyglycoside tablet (TPT) were compared. The results showed that compared with the model group, the percentage of Th17 cells in both the FSN capsule group and high-dose CIA group were obviously decreased (P < 0.05). The percentage of Treg cells had the tendency of increasing in all treatment groups with no significant difference (P > 0.05). There was no significant difference on Th17 and Treg cells among each treatment group and TPT group (P > 0.05). Compared with the model group, the high-dose and middle dose FSN can obviously reduce the levels of TNF-α, IL-6 and IL-17A in synovial fluid of rats in each FSN group (P < 0.05). There was no obvious upregulating effect on IL-4 and IL-10 in synovial fluid of each FSN group (P > 0.05). Compared with the TPT group, there were no significant differences on IL-6, TNF-α, IL-17A, IL-4 and IL-10 in each treatment group (P > 0.05). It was concluded that FSN can obviously reduce the percentage of Th17, decrease the secretion of IL-6, TNF-α and IL-17A in synovial fluid. However, it had no obvious effects on the percentage of Treg cells, or the secretion of IL-4 and IL-10 in synovial fluid. The mechanism of FSN capsule in RA treatment may be through regulating Th17 cells, adjusting body immune imbalance, and inhibiting the excessive secretion of inflammatory media in synovial fluid.

OBJECTIVETo observe the three-dimensional distribution of vessels, and to establish a new method for measurement of blood flow velocity in mice cerebral cortex using two-photon laser scanning microscopy and fluorescence probe labeling technique.

METHODSThe mouse was made cranial window surgery and injected Texas-Red through tail vein after anesthetized. The three-dimensional imaging of vessel was obtained through z-stack scanning, and blood flow velocity was quantified through line scanning.

RESULTSWe could detect vascular distribution for more than 500 µm depth using two-photon microscopy. The velocity of blood flow was (0.59 ± 0.12) mm/s in capillary.

CONCLUSIONThe method for observing the brain blood flow by two-photon microscopy was established, which could achieve quantification of single vascular blood flow velocity and provide experimental evidence for basic research and medical applications.

Animals ; Blood Flow Velocity ; Brain ; blood supply ; Capillaries ; Cerebrovascular Circulation ; Fluorescent Dyes ; Hemodynamics ; Mice ; Microscopy, Fluorescence

Objective To analyze the feature of B cell epitope of soluble liver antigen(SLA) in autoimmune hepatitis. Methods Twenty-one autoimmune hepatitis(AIH) were enrolled the study. The control groups included 15 primary biliary cirrhosis(PBC) patients, 20 chronic virus hepatitis patients and 20 heaths. The serum incubated with coated one dimension of SLA peptide pools by ELISA for mapping the B cell epitopes. ELISPOT was performed to screen the SLA specific T cell response by observing the secretion of IFN-γ after PBMC simulated. Results Ten serum samples from AIH patients recognized the SLA peptide pools, the frequency was 47.62%, which was higher than PBC group, virus hepatitis group and normal control group, P value was 0.001, <0.001 and <0.001, respectively. The 10 positive serum dominant response to SLA 7 th pool(aa385-441), which laid in the unique no conservative region of the SLA molecular. T cell recognizing regions laid dominantly in first and second pool, the frequency was 30% and 40%, respectively, however the recognized frequency of 7th pool was 10%. Conclusion B cell linear epitopes lie in the sequence aa385-441, which is the unique unconservative region in the high conservative SLA molecular and overlaps with part sequence of T cell epitopes. The results show it is complex and interaction that SLA induced humoral-mediated immunity and cellular immunity to involve in AIH pathogenesis.

Objective To explore the influences of the final concentration and adding time of Cytochalasin-B (Cyt-B) on radiation-induced nucleoplasmic bridges (NPB) in cytokinesis-block assay.Methods Hunan peripheral blood samples were divided into 5 final concentration groups (group 2,4,6,8,10 μg/ml) according to different final concentrations of Cyt-B.Moreover,blood samples were divided into 4 adding time groups (group 0,28,40,44 h) according to different adding times of Cyt-B.Blood samples were irradiated with 0 (sham irradiation) and 2 Gy 60Co-rays in vitro,at a dose rate of 1 Gy/min.A cytokinesis-block assay was carried out to prepare NPB samples.The percentages of mononucleated,binucleated and multinucleated cells,as well as the frequencies of NPB and micronucleus (MN) in binucleated cells were analyzed using an optical microscope.Results Nuclear division index (NDI) and the percentages of binucleated cells increased with increased concentration of Cyt-B,and decreased with delayed adding time of Cyt-B (except group 0 h) in both final concentration groups and adding time groups.After exposed to 2 Gy,NPB frequencies were no significant difference (except group 0 h).MN frequencies had the trend of decreased with the increased concentration of Cyt-B,but no significant difference with adding time of Cyt-B.Conclusions In cytokinesis-block assay,different final concentration and adding time of Cyt-B may induce to the variation of NPB frequencies,but there was no significant difference.Appropriate increased final concentration or ahead adding time of Cyt-B can increase the percentage of binucleated cells that help to improve the efficiency of analysis.

Objective To explore the responses of antigen-specific T cells stimulated by hepatitis B virus(HBV)-specific proteins in chronic hepatitis B patients accepting antiviral therapy. Methods Seventeen patients with chronic hepatitis B (CHB) accepting antiviral therapy were included in this study. The peripheral blood monocular cell ( PBMC) were separated from the whole blood collected at the three different time of before and one and three months after accepting antiviral therapy. ELISPOT assay was used to detect the frequency and strength of secreting IFN-γ cells of PBMC stimulated by HBsAg, HBcAg and HBeAg. HBV virus loading, HBsAg, HBeAg, ALT and AST in serum were detected at the same time. Results After three months therapy, ALT, TBiL were improved in all patients, and HBV DNA level were dropped and undetectable in 11 cases. The rates of T cell response in patients to HBV specific proteins were 64. 7% , 76. 5% and 82. 4% at the time of before and one and three months after accepting antiviral therapy, respectively. The frequency of responses of antigen-specific T cells stimulated by HBcAg was higher than that stimulated by HBsAg or HBeAg, and the frequency was enhanced after antiviral therapy. The average response magnitude was expressed as spot forming cells (SFC) per million input cells. SFC of T cell responses to HBcAg was also higher than to HBsAg or HBeAg. There was no significant difference in SFC of T cell responses to HBsAg or HBeAg at the time of before and after antiviral therapy, but there were significant difference in SFC of T cell responses to HBcAg at the time of before and after antiviral therapy. SFC of T cell responses to HBcAg was negatively associated with HBV DNA, and no associated with level of ALT in serum. Conclusion The responses of antigen-specific T cells were improved in CHB patients accepting antiviral therapy which associated with the decrease of HBV DNA. It suggested to investigate HBV specific T cell responses was important.

Adult ; Aged ; Bone Nails ; Female ; Femoral Fractures ; surgery ; Femur ; surgery ; Fracture Fixation, Intramedullary ; Fractures, Comminuted ; surgery ; Humans ; Internal Fixators ; Male ; Middle Aged

Clinical study of modified Ganmai Dazao decoction in the treatment of yang deficiency climacteric depression and observe the effects of modified Ganmai Dazao decoction on neuroendocrine system in patients with yang deficiency climacteric depression. 86 cases were randomly divided into treatment group treated with modified Ganmai Dazao decoction and control group treated with Deanxit. The curative effect was evaluated with Hamilton's depressive scale (HAMD) and pittsburgh sleep quality scale (PSQI) before and at the end of the two and four weeks of the treatment, the serum levels of serotonin (5-HT), norepinephrine (NE), estradiol (E2), follicle stimulating hormone (FSH), luteotropic hormone (LH) were detected before and after the four weeks of the treatment The results showed that the total effective power of treatment group was 88.4% and the total effective power of control group was 81.4% after four weeks interference, with insignificant difference between the two groups. After two and four weeks of the treatment, the score of HAMD decreased remarkably in both groups (P < 0.01), with insignificant difference between the two groups in same phase. After two and four weeks of the treatment, the total score of PSQI decreased remarkably in both groups (P < 0.05), with significant difference between the two groups after four weeks (P < 0.01). After four weeks of treatment, the serum levels of 5-HT and NE increased (P < 0.01), with insignificant difference between the groups. After four weeks of treatment, the serum levels of E2 increased obviously (P < 0.05), the levels of FSH decreased obviously (P < 0.05), the levels of LH decreased insignificant, with insignificant difference between two groups. This study indicates that modified Ganmai Dazao decoction has obvious therapeutic effects in the treatment of climacteric depression, and showed equivalent efficacy with Deanxit, and modified Ganmai Dazao decoction has better effect on improving the sleep quality in patients than Deanxit, the effect of improved clinical symptoms may be through adjusted levels of 5-HT, NE, E2, FSH and LH of climacteric depression.
Adult ; Depression ; blood ; drug therapy ; etiology ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Menopause ; drug effects ; psychology ; Middle Aged ; Neurosecretory Systems ; drug effects ; metabolism ; Norepinephrine ; blood ; Phytotherapy ; Serotonin ; blood

The chemical constituents of Safflower injection were isolated and purified by polyamide, silica gel, Sephadex LH-20, ODS column chromatographies and preparative HPLC. As a result, sixteen compounds have been isolated. Based on the spectral data analysis, their structures were elucidated as scutellarin (1), kaempferol-3-O-β-rutinoside(2), hydroxysafflor yellow A(3), rutin (4), coumalic acid(5), adenosine(6), syringoside(7), (3E)-4-(4'-hydroxyphenyl)-3-buten-2-one(8), (8Z)-decaene-4, 6-diyne-1-Oβ-D-glucopyranoside(9), 4-hydroxybenzaldehyde (10), (2E, 8E) -tetradecadiene-4, 6-diyne-1, 12, 14-triol-1-O-β-D-glucopyranoside (11), kaem-pferol-3-O-β-sophorose (12), uridine (13), roseoside (14), cinnamic acid (15), and kaempferol (16). Compounds 1,2,7,9,11 and 12 were isolated from the Safflower injection for the first time. The anti-platelet aggregation activities of the isolated compounds were assayed. The results indicated all tested compounds exhibited potent activity except for 5, while 2, 3, 9 and 12 showed strong activity against platelet aggregation.
Animals ; Blood Platelets ; drug effects ; physiology ; Carthamus tinctorius ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Fibrinolytic Agents ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Platelet Aggregation ; drug effects ; Rabbits ; Spectrometry, Mass, Electrospray Ionization

Objective:To explore the epidemiological characteristics and the antibiotic resistance of Streptococcus pneumoniae isolates, and to provide the evidence for the rational use of antimicrobial agents to treat Streptococcus pneumoniae infection. Methods:The positive microbiological laboratory identification and antimicrobial susceptibility testing of Streptococcus pneumoniae from sputum of children with respiratory infections during January 2010 to December 2017 in Children′s Hospital of Soochow University were retrospectively analyzed. The positive rates of Streptococcus pneumoniae of different genders, ages, years and seasons were compared. The annual detection rates and trends of drug resistance of Streptococcus pneumoniae to penicillin, amoxicillin and cefotaxime were analyzed by Mann-Kendall trend test. The seasonal decomposition of time series was conducted to assess the association between Streptococcus pneumoniae detection rate and season. Enumeration data was compared using χ2 test. Results:Of the 88 480 sputum specimens, the total positive rate of Streptococcus pneumoniae was 10.3%(9 081/88 480). The detection rates of Streptococcus pneumoniae in children aged 0 to <0.5 years old, 0.5 to <2 years old, 2 to <3 years old, 3 to <5 years old, and 5 to <15 years old were 4.2%(1 407/33 224), 13.1%(3 191/24 390), 14.9%(2 417/16 252), 17.9%(1 474/8 246) and 9.3%(592/6 368), respectively. The difference was statistically significant ( χ2=2 421.6, P<0.01). The detection rates were 8.1%(1 321/16 306) from January to March, 10.9%(2 194/20 207) from April to June, 8.5%(2 141/25 058) from July to September, and 12.7%(3 425/26 909) from October to December. The discrepancy of positive rates in different seasons showed statistical significance ( χ2=311.5, P<0.01). During 2010 to 2017, significant decreases in antibiotic resistant rates of Streptococcus pneumoniae to penicillin, amoxicillin and cefotaxime were detected (tau=-0.93, -0.93 and -0.71, respectively, all P<0.05). Conclusions:The detection rate of Streptococcus pneumoniae in sputum of children with respiratory infections may present seasonal pattern and vary between different ages of children. The resistance to β-lactam antibiotics has declined.

AIMTo study the effects of hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) on proliferation and migration of bovine coronary artery endothelial cells (BCAEC) in vitro.

METHODSBCAECs were isolated and cultured in vitro, and divided into control group, VEGF group and HGF group. BCACEs proliferation were measured using MTT, and their migration was observed using reverse microscope.

RESULTSThe OD value of control, VEGF and HGF group were 0.22 +/- 0.01, 0.40 +/- 0.14, 0.44 +/- 0.15 respectively. The proliferation ratio of BCAECs in VEGF and HGF group was 81.8% +/- 16.9%, 100.0% +/- 21.1% respectively. There was no migration in control group, but significant migration in VEGF and HGF group.

CONCLUSIONBoth VEGF and HGF can promote proliferation and migration of BCAECs, the effect of HGF is stronger than VEGF.

Animals ; Cattle ; Cell Division ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Coronary Vessels ; cytology ; Culture Media, Conditioned ; Endothelial Cells ; drug effects ; Endothelium, Vascular ; cytology ; Hepatocyte Growth Factor ; pharmacology ; Vascular Endothelial Growth Factor A ; pharmacology