AIMTo apply stochastic resonance algorithm (SRA) to quantitative analysis of weak chromatographic signal, which was embedded in the noise.

METHODSBased on the theory of stochastic resonance (SR), a simple and effective SRA has been established to improve analytical detection limits of chromatographic analysis, which apply to enhance the signal to noise ratio by the optimization of the parameters and Runge-Kutta method, was established. The method was used to quantitative analysis of phenazopyridine in human plasma by HPLC/UV. Meanwhile this method is compared with HPLC/MS.

RESULTSBy experimental chromatographic data sets, an excellent quantitative relationship between concentrations of phenazopyridine and their responses had been obtained. The concentration of phenazopyridine in plasma determined by HPLC/UV with SRA and HPLC/MS showed that there was no significant difference (P > 0.05) between the two methods.

CONCLUSIONThe new method was feasible.

Chromatography, High Pressure Liquid ; methods ; Humans ; Mass Spectrometry ; Noise ; Phenazopyridine ; blood ; Sensory Thresholds ; Spectrophotometry, Ultraviolet ; Stochastic Processes

OBJECTIVETo investigate whether cysteinyl leukotriene receptor 1 (CysLT₁ receptor) is involved in rotenone-induced injury of PC12 cells.

METHODSAfter 24 h treatment with rotenone or with rotenone and the CysLT₁ receptor antagonist montelukast, PC12 cell viability was determined by the colorimetric MTT reduction assay. After PC12 cells were treated with various concentrations of rotenone for 24 h or with 3 μmol/L rotenone for various durations, the expression of CysLT(1) receptor was determined by Western blotting, and its intracellular distribution was detected by immunocytochemistry.

RESULTSRotenone (0.3-30 μmol/L) induced PC12 cell injury; this injury was significantly attenuated by montelukast at 1 and 5 μmol/L.The expression of CysLT(1) receptor increased after rotenone treatment at 1-10 μmol/L, or at 3 μmol/L for 3 and 24 h. Rotenone caused concentration-and time-dependent translocation of CysLT₁ receptor from the nucleus to the cytosol.

CONCLUSIONCysteinyl leukotriene receptor 1 is involved in rotenone-induced injury of PC12 cells.

Animals ; PC12 Cells ; Rats ; Receptors, Leukotriene ; metabolism ; physiology ; Rotenone ; toxicity

OBJECTIVETo prepare and identify a polyclonal antibody (pAb) against (mouse) cysteinyl leukotriene receptor 1 (CysLT(1)) and to investigate the changes of CysLT(1) receptor expression in BV2 microglial cells after rotenone treatment.

METHODSRabbits were immunized with KLH-coupled CysLT(1) peptide to prepare the pAb. The titer of the pAb in rabbit plasma was detected by ELISA method, and the specificity of the pAb was tested by antigen blockade. After BV2 cells were treated with rotenone (0.01-1 μmol/L) for 24 h, the expression of CysLT(1) was determined by immunostaining, Western blotting and RT-PCR.

RESULTThe pAb showed a titer of 1/32728, and was not cross-reacted with antigens of CysLT(2) receptor and GPR17. Immunostaining, Western blotting and RT-PCR analysis showed the expression of CysLT(1) receptor in BV2 microglia. Rotenone at 1μmol/L significantly induced an increased expression of CysLT(1) receptor.

CONCLUSIONThe prepared CysLT(1) receptor polyclonal antibody has a high titer and high specificity to meet testing requirements of Western blotting and immunostaining; CysLT(1) is associated with rotenone-induced injury of BV2 microglial cells.

Animals ; Cells, Cultured ; Male ; Mice ; Microglia ; drug effects ; metabolism ; pathology ; Rabbits ; Receptors, Leukotriene ; immunology ; metabolism ; Rotenone ; pharmacology

OBJECTIVETo observe the histopathological finding of bone remodeling in rabbit sinusitis model at different time and the tendency, and to discuss the effect of bone in the pathogenesis of sinusitis.

METHODSFirst, the rabbit sinusitis model was made, then the experimental animals were divided into 3 groups according to the time of infection. There were 8 rabbits in each experimental group, and 4 rabbits in the control group. The sinus specimen were collected, embedded and stained. The bone in the inoculating side and noninoculating side was scored, and the bone in inoculating side was evaluated quantitatively and semiquantitatively. The parameters included the thickness of mucosa, mucoperiosteum, the density of osteoblast, the amount of osteoclast.

RESULTSThe average bone score in the inoculating side was 2.250, 2.875, 2.875; in the noninoculating side was 1.625, 2.250, 2.500. Between group A and the control group, the difference of all three parameters had statistical significance. Between group B and group A, the difference of the thickness of mucosa and the density of osteoblast had statistical significance. Between group C and group B, none of the three parameters had statistical significance.

CONCLUSIONSBacterial sinusitis can lead to bone remodeling, obvious bone destroy can occur at the early phase, then the bone proliferation follows. These results demonstrate that bone remodeling is one of the basic histopathological characters of CRS and might be the reason to lead CRS to a constant and chronic process of inflammation.

Animals ; Bone Remodeling ; Disease Models, Animal ; Inflammation ; Nasal Bone ; pathology ; Rabbits ; Sinusitis ; pathology

BACKGROUNDBiofilms have given new insights to the understanding of pathogenesis of chronic rhinosinusitis (CRS). However, the link between biofilms formation and local inflammatory response remains poorly defined in CRS with nasal polys. The aim of this study was to determine the potential association of the presence of biofilms in the nasal mucosal tissues with clinical features in Chinese patients, which had CRS with nasal polyps (CRSwNP).

METHODSA total of 19 patients with CRSwNP and 12 patients with non-CRS were subjected to endoscopic surgery and their nasal mucosal tissue specimens were examined histologically and by scanning electron microscopy (SEM). Their demographic and clinical features were recorded.

RESULTSThirteen (68.4%) out of the 19 specimens from patients with CRSwNP, but none from control patients, were positive for biofilms that displayed typical characteristics of bacterial and fugal structures. The presence of biofilms in the nasal mucosal tissues was associated with significantly greater values of purulent nasal discharge and preoperative Lund-Kennedy scores, higher levels of serum total IgE and percentages of subjects with endoscopic surgery (ESS) history in patients with CRSwNP, and more severe inflammation in the nasal mucosal tissues of patients with CRSwNP.

CONCLUSIONOur study demonstrated the presence of biofilms in the nasal mucosal tissues of many patients, contributing to the understanding of the pathogenic process of CRSwNP in Chinese patients.

Adolescent ; Adult ; Biofilms ; Chronic Disease ; Female ; Humans ; Male ; Microscopy, Electron, Scanning ; Middle Aged ; Nasal Mucosa ; pathology ; Nasal Polyps ; etiology ; microbiology ; pathology ; Rhinitis ; etiology ; microbiology ; pathology ; Sinusitis ; etiology ; microbiology ; pathology

OBJECTIVETo evaluate the histopathologic changes of the ethmoid bone in CT scan typing of chronic rhinosinusitis (CRS), and investigate relations with outcomes of endoscopic surgery.

METHODSOne hundred and twelve random CRS patients undergoing endoscopic sinus surgery at Tongren hospital were prospectively evaluated. According to the preoperative CT scan, these patients were divided into three groups of CT scan typing. The bony changes and bone remodeling of ethmoid bone were graded and relations with outcomes of endoscopic surgery were investigated.

RESULTSThe CT scan typing results of ethmoid sinus of CRS were as followed: 22.3% (type III), 39.3% (type II), and 38.4% (type I). Comparison of bone remodeling and postoperative endoscopic scores between the three groups revealed a significant difference (P = 0.01, P = 0.02). A trend toward more advanced bone remodeling grade in association with a higher postoperative endoscopic scores was identified, with a statistically significant correlation.

CONCLUSIONThe Chinese CT scan typing of CRS has its pathological basis and can a useful method to predict the surgical outcomes of CRS. The new bone formation or bony tissue remodeling of sinus often represents pathogenesis of CRS and implicates the prognosis of sinus surgery.

Adult ; Chronic Disease ; Ethmoid Sinus ; diagnostic imaging ; pathology ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Prospective Studies ; Sinusitis ; classification ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed ; Young Adult

Objective:To construct the recombinant DNA E.coli ghosts (EBGs) expressing Treponema pallidum adhesin Tp0751 (pcD/Tp0751-BG) and determine its immunocompetence in immunized mice,and provide a potential novel method for syphilis vaccine developing.Methods:The recombinant eukaryotic expression plasmid pcDNA3.1(+)/Tp0751 was constructed and loaded into empty EBGs to create pcD/Tp0751-BG.The loading rate was determined accordingly.Macrophages cell line RAW264.7 was transfected with pcD/Tp0751-BG,and the expression of recombinant Tp0751(rTp0751) protein was detected by Western blot(WB).For immuno-competence in mice,the female BALB/c mice were randomly divided into 6 groups,including three control groups,A (PBS),B (EBG),C (empty pcDNA),and experimental group D(naked pcD/Tp0751),E (pcD/Tp0751-BG) and F (pcD/Tp0751-BG+rTp0751).All the mice were immunized as indicated for three times by intramuscular injection at two weeks intervals.The levels of specific IgG in sera and SIgA in genital tract lavage fluid were measured by ELISA.Levels of lymphocyte proliferation and IFN-γ secretion in spleen cells were measured by CCK-8 Cell Counting Kit and ELISA as well,respectively.Results:The loading rate of pcD/Tp0751 to EBGs was 76.1%.WB showed that the target recombinant protein pcD/Tp0751 expressed in RAW264.7 cells was active with Tp-infected rabbit sera.The titers of specific IgG and SIgA in group D,E,F gradually increased to significantly higher level as compared to the control groups (P<0. 01),which reached its peak at wk 8 after last immunization(the titers of IgG and SIgA were 1 :102 400 and 1 :12 800 in group F,respectively). Higher levels of specific IgG and SIgA were observed in groups E and F as compared to group D after first boost (P<0. 01),with groups F higher than group E after last boost(P<0. 01). At wk 8 after the last boost,the stimulation index (SI) and levels of IFN-γ in group D,E,F were all significant higher than the control groups (P<0. 01), with group E and F higher than group D (P<0. 01),and group E higher than group F (P<0. 05). Conclusion: The recombinant DNA EBGs of T. pallidum adhesin Tp0751 (pcD/ Tp0751-BG) possesses the immunocompetence to induce not only strong mucosal and systemic humoral immune response but also systemic cellular immune response in BALB/ c mice. The heterologous boost can be more efficient than homologous boost during immunization process.

Animals ; Female ; Ginsenosides ; pharmacology ; therapeutic use ; Hepatocytes ; drug effects ; metabolism ; ultrastructure ; Liver Cirrhosis, Experimental ; drug therapy ; metabolism ; Male ; Phytotherapy ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

BACKGROUNDAdenoid hypertrophy (AH) is associated with pediatric chronic rhinosinusitis (pCRS), but its role in the inflammatory process of pCRS is unclear. It is thought that innate immunity gene expression is disrupted in the epithelium of patients with chronic rhinosinusitis (CRS), including antimicrobial peptides and pattern recognition receptors (PRRs). The aim of this preliminary study was to detect the expression of innate immunity genes in epithelial cells of hypertrophic adenoids with and without pCRS to better understand their role in pCRS.

METHODSNine pCRS patients and nine simple AH patients undergoing adenoidectomy were recruited for the study. Adenoidal epithelium was isolated, and real-time quantitative polymerase chain reaction (RT-qPCR) was employed to measure relative expression levels of the following messenger RNAs in hypertrophic adenoid epithelial cells of pediatric patients with and without CRS: Human β-defensin (HBD) 2 and 3, surfactant protein (SP)-A and D, toll-like receptors 1-10, nucleotide-binding oligomerization domain (NOD)-like receptors NOD 1, NOD 2, and NACHT, LRR and PYD domains-containing protein 3, retinoic acid-induced gene 1, melanoma differentiation-associated gene 5, and nuclear factor-κB (NF-κB). RT-qPCR data from two groups were analyzed by independent sample t-tests and Mann-Whitney U-tests.

RESULTSThe relative expression of SP-D in adenoidal epithelium of pCRS group was significantly lower than that in AH group (pCRS 0.73 ± 0.10 vs. AH 1.21 ± 0.15; P = 0.0173, t = 2.654). The relative expression levels of all tested PRRs and NF-κB, as well as HBD-2, HBD-3, and SP-A, showed no statistically significant differences in isolated adenoidal epithelium between pCRS group and AH group.

CONCLUSIONSDown-regulated SP-D levels in adenoidal epithelium may contribute to the development of pCRS. PRRs, however, are unlikely to play a significant role in the inflammatory process of pCRS.

Adenoids ; cytology ; Antimicrobial Cationic Peptides ; metabolism ; Child ; Epithelial Cells ; metabolism ; Female ; Humans ; Immunity, Innate ; genetics ; physiology ; Male ; Receptors, Pattern Recognition ; metabolism ; Sinusitis ; metabolism ; Toll-Like Receptors ; metabolism