Objective To study the role of TLR2 and TLR4 signal transduction in RAW264.7 monocyte-macrophages stimulated by Aspergillus fumigatus conidia,and to investigate the expression of TLR2 signal transduction after silencing gene of TLR4.Methods Macrophages were randomly divided into normal group ( N group),normal+stimulated with Aspergillus fumigatus conidia ( N +Af group ),normal + transfected with TLR4-siRNA [ TLR4 (RNAi) group ],normal+transfected with TLR4-siRNA +stimulated with Aspergillus fumigatus conidia[ TLR4(RNAi) +Af group].RT-PCR and Western blot were used to assay expression levels of TLR2,TLR4,MyD88 mRNA and pro-inflammatory cytokines TNF-α protein when macrophages were stimulated 12 h by Aspergillus fumigatus conidia after tranfected 24 h with TLR4-siRNA by technology of RNAi.Results ( 1 ) Compared with N group,the expression of TLR2,TLR4,MyD88 mRNA and TNF-αprotein in N+Af group significantly increased before silencing gene of TLR4.(2) Silencing efficiency of macrophates was up to 83％ after transfected with TLR4-siRNA.(3)The expression of TLR2,MyD88 mRNA in TLR4 (RNAi) group significantly decreased contrast with normal group.Meanwhile the expression of TLR2,MyD88 mRNA and TNF-α protein also obviously reduced in TLR4(RNAi) +Af group when compared with N +Af group.Compared with TLR4 (RNAi) group,the expression of MyD88 mRNA in TLR4 (RNAi) +Af group significantly increased.However,the expression of TLR2 mRNA and TNF-α protein have no significant change after silencing gene of TLR4.Conclusion Signaling pathway of TLR2 and TLR4 in macrophages was activated by given stimulus of Aspergillusfumigatus conidia and exerted the effect of anti-Aspergillus fumigatus spores stimulation through the release of pro-inflammatory cytokines TNF-α.Meanwhile,silencing gene of TLR4 down-regulate the effect of TLR2 signal transduction in RAW264.7 cells to anti-Aspergillus fumigatus conidia stimulation,and it found that TLR4 played an more important role by contrast with TLR2.

Objective To assess effects of stress dynamic CT myocardial perfusion imaging (CT-MPI) combined with coronary CT angiography (CCTA) on the diagnosis of myocardial perfusion defects in coronary artery disease (CAD). Methods Patients with CAD diagnosed by CCTA underwent ATP stress CT-MPI examination. Single-photon emission computed tomography (SPECT) myocardial perfusion imaging (SPECT-MPI) was performed within one week and set as the reference standard. CT-MPI results were qualitatively analyzed, and myocardial blood flow (MBF), myocardial blood volume (MBV) as well as time to peak (TTP) were quantified according to CT-MPI. Effects of CCTA, CT-MPI, and CT-MPI combined with CCTA on predicting myocardial perfusion defects were assessed in comparison with NMPI. Results Thirty patients [(54.8±8.4)years] were enrolled in our study, 20 were men (68%). MBF [(79.3±18.0) versus (135.1± 35.2) ml·100 ml-1·min-1] and MBV [(8.9±2.9) versus (13.8±8.9) ml/100 ml] were significantly decreased in hypoperfused segments compared with normal segments, while TTP was increased in hypoperfused segments [(13.9 ± 2.5)s] compared with normal segments [(9.1 ± 2.1)s] (t=0.302, 0.866 and 0.024 respectively, all P values<0.01). The sensitivity, specificity of CT-MPI for identifying segments with perfusion defects were 91.3%(147/161), 84.6%(281/332), respectively. On a per-vessel basis, the area under the receiver operating characteristic curve for predicting myocardial perfusion defects were 0.635(95%CI:0.517—0.753) for CCTA, 0.709(95%CI:0.599—0.819)for CT-MPI, and 0.837(95%CI:0.749—0.925)for CT-MPI combined with CCTA, respectively. Conclusions The performance of stress dynamic CT-MPI in the diagnosis of myocardial perfusion defects in CAD was good. One-stop examination of CT-MPI combined with CCTA improves the diagnostic accuracy for identifying flow-obstructing stenosis.

OBJECTIVE: To standardize and simplify the drug receving and preparing model. METHODS: Reasonable pharamcy inventory parameters were set up based on our experience; second order drug storeroom dynamic inventory system was designed and applied, and the indexes of drug storeroom inventory and workload of the new and the old models were reported and analyzed. RESULTS: The new warning system provided an optimal inventory, and in which the incidences of overstock or stock shortage, the frequency of daily drug receving, drug kinds and the time used with checking stock and filling receipt were all significantly less than in the traditional model. CONCLUSION: Due to the second order drug storeroom dynamic inventory warning system, the drug receiving was more scientific, the drug stock amount was reduced and the drug receiving time was saved, and the mangament level of hospital drug storeroom inventory was enhanced.

Objective To explore the mechanism of anti-tumor effects of transferring tumor-specif-ic lymphocytes obtained from pre-immunized BALB/c mice with inactive rolL-21 tumor vaccine (mIL-21-Sp2/0)to syngeneic mice, associated with mIL-21 tumor vaccine immunization, in the condition of cyclo-phosphamide (Cy)-induced lymphopenia. Methods Activated lymphocytes of spleen and lymph nodes ob-tained from pre-immunlzed syngeneic mice with irradiated mIL-21-Sp2/0 cells were infused into BALB/cmice treated with Cy 2 days before, subsequently vaccinated with mlL-21 tumor vaccine, after 7 days, chal-lenged with Sp2/0 tumor cells, observed the growth of tumor of mice. T lymphocyte subsets differentiation was measured by flow cytometry (FCM) analysis. The proliferation and cytotoxie activities of activated lym-phocytes were analyzed by FCM, respectively, staining with CFSE and 7-AAD. The number of IFN-γ-secre-ting cells was evaluated by ELISPOT. Results The lymphopenic mice were transferred with activated lym-phocytes and inoculated with raiL-21 tumor vaccine might provide superior anti-tumor immunoprotection, re-tard tumor growth of the mice. The proliferating capabilities and killing rate of transferred tumor Ag-specific lymphocytes enhanced obviously, the number of IFN-γ-secreting cells was significantly higher compared with the control groups. Conclusion Under Cy-induced lymphopenia condition, tumor Ag-specific lymphocytes sensitized by raiL-21 tumor vaccine were transferred to mice and immunized with mlL-21 tumor vaccine at the same time, benefit the proliferation of transferred effective cells and immune cells itself, assist to form and sustain special anti-tumor effects.

Objective:To establish the RPB5-mediating protein (RMP)-silenced stable cell lines and study the inhibitory effects of small interfering RNA (siRNA) targeting RMP gene on the proliferation and migration of human hepatoma SMMC-7721 cells. Methods:Three RMPi siRNAs were designed and synthesized in vitro and transfected into SMMC-7721 cells. The inhibitory effect of siRNA on RMP gene expression was measured by RT-PCR to select the best siRNA. The expression vector pGPU6-Neo-RMP-484 was transfected into SMMC-7721 cells by the lipofectamine and the cells stably expressing the siRNA were selected by G418. RT-PCR was used to detect the interference efficacy against RMP gene. Cell proliferation and adhesion were measured by MTT assay. Wound healing test was used to observe the migration ability of cells. Results:The SMMC-7721 cell lines with down-regulated RMP expression were established by using RNA interference technology. Compared with the negative control cells, expression of RMP mRNA was down-regulated by(83.67±2.56)% .The proliferation of stable-transfected cells was inhibited by(74.33±0.58)% . The adhesion capability of stable-transfected cells was enhanced but the migration capacity was decreased compared with the negative control cells. Conclusion:The pGPU6-Neo-RMP-484 cell lines with stable transfection of RMP siRNA recombinant vector are successfully screened,which can be used as a cellular model for studying the molecular mechanism of RMP. Down-regulation of RMP gene expression can effectively inhibit the proliferation, enhance the adhesion, and decrease the migration of SMMC-7721 cells.

Objective:To establish the quality standard for Tangzhixiao capsules. Methods: The qualitative identification of Salvia miltiorrhiza,Atractylodes and Hawthorn was detected by TLC, and the quantitative determination of salvianolic acid B was determined by HPLC. The HPLC determination was performed on a YMC-Triart C18 (250 mm × 4. 6 mm, 5μm ) column with the mobile phase consisted of acetonitrile-0. 1% phosphoric acid (24∶ 76) at the flow rate of 1. 0 ml·min-1 , the column temperature was 30℃ and the detection wavelength was set at 286 nm. Results: The TLC spots were fairly clear, and the negative samples showed no interference. The concentration of salvianolic acid B within the range of 0. 012-0. 120 mg·ml -1 was linear with peak area(r = 0. 999 5), the average recovery was 100. 06% and RSD was 0. 83%(n = 6). Conclusion: The method is accurate, reliable and reproducible, which can be used in the quality control of Tangzhixiao capsules.

A method for quantitative detection of florfenicol by colloidal gold lateral flow immunoassay was developed.The experimental conditions including pH value, concentrations of antibody in the process of conjugation between the colloidal gold and antibody, amount of gold-labeled antibody, concentration of the antigen sprayed on test lines (T line), and detection time were optimized.With a colloidal gold strip reader, the signal intensity of T lines and control lines (C line) on lateral flow strips was recorded.The T/C ratio of negative control and positive samples was defined as B0 and Bx, and the standard curve was established by plotting the Bx/B0 ratio against the concentration of florfenicol.This assay showed a good linear range from 0.1 to 1.5 ng/mL with the limit of detection of 0.08 ng/mL, while the result could be obtained within 15 min.The result showed that this quantitative method was convenient and rapid, and could be used in screening a large amount of samples on site.

can specifically combined with EGFR, which may be applied to noninvasive NIRF imaging of tumors highly expressed EGFR in vivo.

OBJECTIVETo prepare berberine microemulsion, and to investigate its properities and the absorption character in rat intestine in situ.

METHODThe optimum formulation of the blank microemulsion selected by pseudo tertiary phase diagrams and the berberine microemulsion was prepared based on the blank microemulsion. The viscosity, conductance, refraction rate and particle size of berberine microemulsion were surveyed. An in situ rat perfusion method was used to investigate the intestinal absorption of berberine microemulsion. A UV method for determination of berberine in the intestinal flux was established.

RESULTThe viscosity, conductance, refraction rate and particle size of berberine microemulsion were 2.11 cPas, 125.5 microomega, 1.363 and 24.0 nm, respectively. The absorption rate of berberine at the ileum was the best. The absorption of berberine microemulsion at the ileum was significantly higher than that of raw medicine (P < 0.01).

CONCLUSIONThe microemulsion system might improve the absorption of berberine in the intestinal tract.

Absorption ; Administration, Cutaneous ; Animals ; Berberine ; administration & dosage ; pharmacokinetics ; Drug Delivery Systems ; Drug Stability ; Female ; Ileum ; metabolism ; Intestinal Absorption ; drug effects ; Intestines ; metabolism ; Male ; Particle Size ; Rats ; Rats, Wistar ; Skin ; metabolism ; Skin Absorption ; drug effects ; physiology ; Solubility ; Technology, Pharmaceutical ; methods

Objective To explore the management of spontaneous intraspinal hematoma.Methods From January 2011 to July 2018,29 cases with spontaneous intraspinal hematoma were admitted to our department.Date on etiology,clinical presentation,radiological features,treatment strategy and prognosis were analyzed retrospectively.The prognosis was assessed by American Spinal Injury Association impairment scale (ASIA) before and after the treatment.Results Total of 29 cases,only 10 cases (34.5％) revealed specific etiology,including 7 cases of spinal vascular malformation,2 of tumor apoplexy,1 of cavernous hemangioma.After 2 weeks of conservative treatment,3 patients with grade D and 3 patients with grade E were assessed for spinal function.The average interval from onset to surgery was(9.4±7.5) days,the ASIA after two weeks of the operation was as follows:5 patients were assessed at grade A,5 patients at grade C,8 patients at grade D and 4 patients at grade E.28 patients were followed up for (48.7±23.1) months on average,6 patients without surgery were E,22 cases with surgery were as follows:4 cases A,18 cases D/E.Conclusions The etiology of spontaneous intraspinal hematoma is hard to define even after complete preoperative examination and exploratory operation.The preoperative neurologic functions are important predicting factors for the prognosis of spontaneous intraspinal hematoma.For patients who had neurologic function deficit,surgical treatment should be performed urgently to remove the hematoma and release the decompression of spinal cord.The majority of these patients can achieve a positive prognosis after surgery.