Objective To investigate the immunity effects of quorum sensing in the pulmonary infection in rats due to Pseudomonas aeruginosa.Method(1)300 healthy,clean Sprague-Dawley rats were divided into 3 groups randomly:two different Pseudomonas aeruginosa(the wide-type Pseudomonas aeruginosa PAO1 and its double mutant PAO-JP2,in which the quorum sensing systems were defective,embedded in minute seaweed algiante beads which was made by an ejection set with an acuminate hole were inoculated to Sprague-Dawley rats to establish animal model of chronic pulmonary infection.The control group were dealed with the same methods using the sterile brine instead.(2)The levels of antibody IgG,IgM,IgG1,IgG2a to Pseudomonas aeruginosa in sera and cytokines including interferon-?(IFN-?),Interleukin-4(IL-4)in lung homogenate was measured at 3,7,14,28 days after infection.Results(1)The mortality in the PAO1-JP2-infected group(11.0%)was significantly lower than that of the PAO1-infected group(29.7%)and the control group(4.21%).(2) Compared with the PAO-JP2 group,during the early stages of infection(3 days after infection),the IFN-?level in lung homogenates was significantly higher;during the middle stages of infection(7 days after infection),the levels of IFN-?and IL-4 of PAO1-infected rats were significantly higher;In the later stages of infection(14 to 28 days after infection),levles of IFN-?,IgG2a were lower,while levels of IL-4,IgG,IgG1 were higher persistantly in PAO1-infected rats.Conclusions Quorum sensing system play an important role in pathogenesis and immunity effects of pseudomonas aeruginosa infection via modulating the expression of virulence factors and interfering with the immune system of hosts.

OBJECTIVETo develop a safe and effective lentivirus vaccine model and provide insights into the development of other lentivirus vaccines.

METHODSIn this study, a construct of pGPT was made by deleting env gene in the infectious Equine infectious anemia virus (EIAV) molecular clone of WU57. Since the overlaping of EIAV Rev gene with env gene, there was no Rev gene in the construct of pGPT. For compensation of Rev function, the construct of pGPTC was made by inserting 4 copies of constitutive RNA transport elements (CTEs) from Mason-Pfizer monkey virus into the construct of pGPT. In addition, a construct designated pTEB expressing EIAV Env protein was made while env gene-minus viruses were made by co-transfection of pGPT/pTEB or pGPTC/pTEB into 293 cells. Western blot was used to identify the development of recombinant virus particles. Then immunofluorescence assay was used to evaluate the infectivity of recombinant virus particles in vitro.

RESULTSEIAV proteins expression was detected in the supernatant of transfected 293 cells by Western blot within pGPTC/pTEB transfected cells. However, no evidence of EIAV proteins expression was observed within pGPT/pTEB transfected cells. EIAV proteins expression was detected in the first round but not in the second round infected EK cells with EIAV(GPTC) by immunofluorescence assay.

CONCLUSIONRev/RRE was necessary for expression of viral structural proteins; CTEs from Mason-Pfizer monkey virus was functionally interchangeable with EIAV Rev/RRE to help RNAs transportation out of nucleus to express structural proteins and EIAV particles were produced in the transfected 293 cells. A live EIAV recombinant virus with single round infection had been developed.

Animals ; Blotting, Western ; Cells, Cultured ; Fluorescent Antibody Technique ; Genes, rev ; Haplorhini ; Horses ; Humans ; Infectious Anemia Virus, Equine ; genetics ; Lentivirus ; immunology ; Lentivirus Infections ; immunology ; prevention & control ; Mason-Pfizer monkey virus ; genetics ; Transfection ; Vaccines, Synthetic ; Viral Vaccines ; genetics ; immunology

Objective To investigate the effect of glucocortieoids on tenasein C(TNC) expression in nasal polyp tissues and airway epithelia,and explore the possible mechanism of glucocorticoids inhibiting remodeling of nasal polyp tissue.Methods The enzyme linked immunosorbent assay(ELISA) was used to detect the protein levels of tenascin C and transforming growth factor-β1(TGF-β1) in nasal polyp tissues from intranasal glucocorticoids (Budesonide,BUD) treated group and untreated group.The cell culture,real-time RT-PCR and in situ ELISA were employed to investigate the regulatory effects of budesonide on the TNC mRNA and protein expression in BEAS-2B immortalized human bronchial epithelial cells.Results The protein levels of TNC and TGF-β1 were decreased in nasal polyp tissues from intranasal BUD-treated group as compared with untreated group(P<0.01).There was a significantly positive correlation between TNC and TGF-β1 protein levels in nasal polyp tissues (r =0.68,P<0.01).After preincubation with BUD,TNC mRNA and protein expression induced by TGF-β1 in BEAS-2B cells was significantly decreased(P<0.01).Conclusion Glucocorticoids might participate in the regulation of tissue remodeling in nasal polyp by inhibiting the TGF-β1 expression in nasal polyp tissue and suppressing the induction of TGF-β1 to up-regulatie the TNC expression in airway epithelia.

Objective To explore the effects of aging on ventricular remodeling and cardiac rupture after acute myocardial infarction in mice. Methods　Male C57BL/6 mice of 3 months and 12 months old were randomly divided into sham operation group and myocardial infarction(MI)group.Following acute myocardial infarction(AMI)modeling induced by open-chest surgery,the events of cardiac rupture were monitored and the echocardiography and hemodynamics were performed on the 7th day after surgery.Zymography,immunohistochemical method and pathological staining were used to measure the activity of matrix metalloproteinases(MMPs),the content of collagen and the degree of inflammatory cell infiltration on the 3rd and 7th days after surgery,respectively. Results　The incidence of cardiac rupture was higher in elderly group than that in young group(38.0% vs.16.0%,X2=6.139,P＜0.05).Compared with young group,significant infarct expansion,left ventricular (LV)remodeling and hemodynamic deterioration were showed in elderly group on the 7th day after surgery(t=5.754,P＜0.05).The degree of inflammatory cell infiltration and the expression of MMP-9 were significantly increased in elderly group on the 3rd day following AMI modeling(P＜0.05),and the collagen content and the expression of type Ⅲ collagen were significantly increased (P＜0.05)compared with young group. Conclusions　Aging is a risk factor for post-infarct cardiac rupture in the mice model.The mechanisms which are responsible for this age-related difference of cardiac rupture are related to increasing degree of inflammatory cell infiltration, overexpression of MMP-9 and type Ⅲ collagen and aggravated early LV remodeling.

A new approach based on online coupled liquid chromatography-gas chromatography-mass spectrometry ( LC-GC / MS ) was developed for the rapid determination of 4-( methylnitrosamino )-1-( 3-pyridyl)-1-butanone (NNK) in mainstream cigarette smoke, in which a switching valve was employed to online switching between two-dimensional chromatography. The online LC-GC / MS system used in this study was built by using online gel permeation chromatography-gas chromatography-mass spectrometry except that the micro gel column was replaced by micro alkaline alumina column which was prepared by ourselves before. The NNK in mainstream cigarette smoke was collected by a Cambridge filter pad, then the pad was extracted with dichloromethane, and the extract was quantitatively analyzed by online LC-GC / MS with D4-NNK as an internal standard. Online LC-GC / MS allowed online pretreatment purification, and the sample was subjected to online LC-GC / MS without any prior purification, which reduced human error in analysis process. The injection volume of the present online LC-GC / MS could reach 40 μL, which was 20 times of that of the conventional GC / MS (2. 0 μL of injection volume), and thus significantly improved the sensitivity. Under the optimum conditions, good linearity ( r = 0. 9998) was obtained over the range of 1. 2 - 120 ng / mL. The recoveries at three spiked levels ranged from 93. 9% to 96. 0% , and the limits of detection for qualitative and quantitative detection were 0. 25 ng / mL and 0. 9 ng / mL, respectively. All the results obtained by the present method are comparable to those of standard method recommend by China National Tobacco Company.

The 1H-NMR fingerprints of three different species tibetan medicine sea buckthorn were established by 1H-HMR metabolomics to find out different motablism which could provide a new method for the quality evaluation of sea buckthorn. The obtained free induction decay (FID) signal will be imported into MestReNova software and into divide segments. The data will be normalized and processed by principal component analysis and.partial least squares discriminant analysis to perform pattern recognition. The results showed that 25 metabolites belonging to different chemical types were detected from sea buckthorn,including flavonoids, triterpenoids, amino acids, carbohydrates, fatty acids, etc. PCA and PLS-DA analysis showed three different varietiest of sea buckthorn that can be clearly separated by the content of L-quebrachitol, malic acid and some unidentified sugars, which can be used as the differences metabolites of three species of sea buckthorn. 1H-NMR-based metabonomies method had a holistic characteristic with sample preparation and handling. The results of this study can offer an important reference for the species identification and quality control of sea buckthorn.
Hippophae ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Medicine, Tibetan Traditional ; Metabolomics

Objective To measure the serum selenium levels in patient with Keshan disease(KSD)and in healthy controls in Shandong,Sichuan and Inner Mongolia KSD areas,to monitor the long-term dynamic changes of hair and serum selenium levels in Shandong KSD areas,and to provide scientific basis for preventing KSD.Methods A cross-sectional survey was carried out in KSD areas of Shandong,Sichuan and Inner Mongolia in 2009.The research subjects which come from KSD areas were 77 cases and 63 healthy controls from Shandong;35 patients and 36 healthy controls from Sichuan;and 17 patients and 18 healthy controls from Inner Mongolia.Additional 33 healthy people from Jinan city were selected as controls of non-KSD areas.Blood and hair samples were collected and selenium levels were measured by 2,3-diaminonaphthalene fluorescence spectrometry.Retrospective method was used to analyze the hair and serum selenium data collected between 1976 and 2004 in Shandong KSD areas.and these data were eompard with the data of 2009 to observe the long-term dynamic changes.Results ① The serum selenium levels of KSD patients in Shandong and Inner Mongolia were significantly lower than that of healthy subjects of KSD areas[(0.0773±0.0113)vs(0.0895±0.0256),(0.0347±0.0107)vs(0.0469±0.0161),t=3.52,3.87,all P<0.01].No significant difference was found between KSD patients and healthy people in Sichuan[(0.0792±0.0162)vs(0.0774±0.0103),t=0.55,P>0.05].②The serum selenium levels of KSD patients in Shandong,Sichuan and Inner Mongolia KSD areas were lower than that of non-KSD area[(0.0988±0.0231)mg/L,q=6.74,5.83,19.47,all P<0.01].The serum selenium levels of healthy people in Sichuan and Inner Mongoha KSD areas were significantly lower than that of non-KSD area(q=6.68,16.36,all P<0.01).The serum selenium levels of healthy controls in Inner Mongolia were lower than that of in Shandong and Sichuan(q=13.63,14.74,13.62,1.46,all P<0.01).③From 1976 to 2009,the hair and serum selenium levels of Shandong resident were increased 1.68 times(0.343/0.128-1)for hair and 0.98 times(0.091/0.046-1)for serum,respectively.But there was no significant difference between the average growth rate of hair and serum selenium levels(χ2=1.38,P>0.05).Conclusions ①The hair and serum selenium levels of KSD patients are lower than that of healthy controls in non-KSD area.②The serum selenium levels of Shandong,Sichuan and Inner Mongolia are different between KSD patients and healthy controls in the diseased areas.③The hair and serum selenium data of Shandong resident show an upward vend over the past 30 years.We suggest to continue the comprehensive measures of adding selenium in KSD areas.

AIMTo prepare rifapentine (RIF) liposomes modified by surfactants for studying their the water-solubility, drug loading effeciency, release rate and pulmonary drug delivery.

METHODSThe film method was used to prepare RIF liposomes. Of verious RIF liposomes morphology by lauric diethanolamide (LDEA), Tween 80 and azone, the properties were studied, envolving morphology, entrapment drug release rate and dissected lung-membrance penetration rate of swine. Pulmonary delivery study was carried out through bronchoscope.

RESULTSThe particle size of RIF-LDEA liposomes was between 15 - 50 nm. The top entrapment efficiency reached 83.0%. The apparent coefficient of membrane penetration (Kp) was 44.29. LD50 was 675 mg x kg(-1) by iv.

CONCLUSIONLDEA increased the water-solubility, loading effeciency and release rate of RIF liposomes. The prepared RIF-LDEA liposomes were suitable for the treatment of pulmonary tubrculosis through bronchoscope.

Animals ; Antibiotics, Antitubercular ; administration & dosage ; chemistry ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; methods ; Ethanolamines ; chemistry ; Lauric Acids ; chemistry ; Liposomes ; Lung ; metabolism ; Particle Size ; Permeability ; Rifampin ; administration & dosage ; analogs & derivatives ; chemistry ; Solubility ; Swine

OBJECTIVE: To construct the expressing vector of siRNA which can inhibit the Smad3 activity. METHODS: Sixty-four bases of 2 pair oligos for hairp in RNA expression which targeted Smad3 gene were chemically synthesized and annealed. pSUPER vector was linearized with BgL II and Hin d III treated with alkaline phosphatase (CIP). Anneled oligos were inserted into the downstream of the treated pSUPER's pol III H1 promoter to construct RNAi plasmid (pSUPER Smad3). Oligos with a scrambled sequence were used as a negative control. pSUPER Smad3 was transfected into human renal tubular epithelial cells (HKC). RESULTS: Recombinant pSUPER Smad3 vector was identified by the digestion with Eco R I and Hin d III, and confirmed by the sequencing analysis with T3 primer. Sixty-four bases had been inserted into the expected site. Furthermore, the insertion sequence was exactly corrected. The activity evaluation indicated that mRNA and protein of Smad3 but not Smad2 were inhibited by pSUPER Smad3 in HKC. CONCLUSION The pSUPER Smad3 system has been constructed successfully, and has high inhibition and specificity in vitro.
Epithelial Cells ; metabolism ; Humans ; Kidney Tubules ; cytology ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Smad3 Protein ; antagonists & inhibitors ; genetics ; Transfection

Objective To investigate the effects of hyperbaric oxygen (HBO) combined with neural stem cell (NSC) transplantation on functional recovery after spinal cord injury (SCI).Methods Forty healthy adult Sprague-Dawley rats were randomly divided into a sham group,an SCI group,an NSC group and an HBO+NSC group.In the latter three groups,complete spinal cord transection was performed at T10.The rats of the HBO+NSC group were treated with HBO therapy after NSC transplantation.Hindlimb motor function was assessed with the BassoBeattie-Bresnahan (BBB) scale and tilt table testing.Motor evoked potentials (MEPs) were measured to evaluate the rats'nerve conduction function.The amount of surviving transplanted NSCs at the injured site was counted using Hoechst dyeing.Results The average BBB scores and tilt table test results in the HBO+NSCs group were significantly better than in the SCI and NSC groups from the 2nd week post-SCI.By the 4th week the average latency period and MEP amplitude in the HBO+NSC group were significantly better than in the NSC group,while MEPs in the SCI group still had not re-appeared.The surviving amount of transplanted NSCs in the HBO+NSC group was also significantly greater than in the NSC group.Conclusion HBO combined with NSC transplantation has synergistic effects and can improve functional recovery after SCI.This may be explained by the important role of HBO in promoting the survival of transplanted NSCs.