RESUMO
Cryptococcus is an opportunistic pathogenic fungus and is the major cause of fungal meningitis. The cryptococcal antigen (CrAg) lateral flow assay (LFA) is an immunochromatographic test system that has simplified diagnosis as a point-of-care test. In this study, we evaluated the diagnostic performance of Cryptococcal capsular polysaccharide detection FungiXpert (Genobio Pharmaceutical, Tianjin, China) using serum and cerebrospinal fluid (CSF) samples for the diagnosis of cryptococcosis and investigated the cross-reaction of the assays to pathogenic fungi and bacterium by comparing it to the U.S. Food and Drug Administration (US FDA)-approved IMMY CrAg LFA. Eighty CSF and 119 serum/plasma samples from 158 patients were retrospectively collected to test for qualitative or semi-quantitative detection of CrAg. Cross-reaction of the assays was tested using 28 fungi and 1 bacterium. Compared to IMMY CrAg LFA, the FungiXpert LFA demonstrated 99.1% sensitivity and 98.9% specificity in the qualitative test. In the 96 semi-quantitative CrAg assay results, 39 (40.6%) test titers of FungiXpert LFA were 1-2 dilutions higher than those of IMMY CrAg LFA. The Intraclass Correlation Coefficient of the Semi-quantitative results of CrAg titer tests via the two assays was 0.976. Similar to IMMY CrAg LFA, FungiXpert LFA showed cross-reactivity with Trichosporon asahii. Compared with the IMMY CrAg LFA, the FungiXpert LFA showed an equal, yet, excellent performance. However, it is important to note that these two assays have potential cross-reactivity to T. asahii when diagnosing patients. FungiXpert LFA is a rapid screening method for the effective and practical diagnosis and treatment of cryptococcosis. LAY SUMMARY: The FungiXpert LFA was developed to diagnose fungal meningitis caused by Cryptococcus yeasts, by using serum or cerebrospinal fluid. It was compared to an existing lateral flow assay (LFA). The FungiXpert LFA performed well in qualitative and semi-quantitative tests.
Assuntos
Criptococose , Cryptococcus , Infecções por HIV , Meningite Criptocócica , Meningite Fúngica , Animais , Antígenos de Fungos , Criptococose/diagnóstico , Criptococose/veterinária , Infecções por HIV/veterinária , Meningite Criptocócica/diagnóstico , Meningite Criptocócica/veterinária , Meningite Fúngica/veterinária , Polissacarídeos , Estudos RetrospectivosRESUMO
To investigated the molecular epidemiology and in vitro antifungal susceptibility of Cryptococcus isolates from West China Hospital from HIV and non-HIV patients between 2009 and 2015. A total of 132 C. neoformans and C. gattii were subjected to antifungal susceptibility testing by E-test method. Among the 132 isolates, 42 C. neoformans and C. gattii were analyzed by mating type and URA5-RFLP. A total of 113 C. neoformans and C. gattii were subjected to multi-locus sequence typing (MLST). MLST results revealed that ST5 was the major molecular type. The wild-type (WT) phenotype was seen in 91.5-100% of C. neoformans isolates for amphotericin B, 5-flucytosine, fluconazole, and voriconazole. However, 72.3% (94/130) of C. neoformans isolates were non-wild-type (non-WT) to itraconazole by E-test method. In the sixth study year, the geometric mean, MIC50 and MIC90 of fluconazole were the highest (P < 0.001). Among 132 patients. 52 were coinfected with HIV and 80 were HIV-negative. Isolates from HIV and non-HIV patients showed no differences in susceptibility to amphotericin B (P = 0.544), 5-flucytosine (P = 0.063), fluconazole (P = 0.570), voriconazole (P = 0.542), and itraconazole (P = 0.787). Our study showed that Cryptococcus in southwest China showed a low degree of genetic diversity. The increased MIC values of fluconazole are noted. Cryptococcus isolates from HIV and non-HIV patients have shown no differences in susceptibility to five antifungal agents.
Assuntos
Antifúngicos/farmacologia , Criptococose , Cryptococcus gattii , Cryptococcus neoformans , Farmacorresistência Fúngica/genética , Infecções por HIV/epidemiologia , Adolescente , Adulto , Idoso , China/epidemiologia , Criptococose/epidemiologia , Criptococose/microbiologia , Cryptococcus gattii/efeitos dos fármacos , Cryptococcus gattii/genética , Cryptococcus neoformans/efeitos dos fármacos , Cryptococcus neoformans/genética , Feminino , Variação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
OBJECTIVE: To determine the value of galactomannan (GM) in bronchoalveolar lavage fluid (BALF) for diagnosing invasive pulmonary aspergillosis. METHODS: According to the European Organization for Research and Treatment of Cancer / Invasive Fungal Infection Group (EORTC / IFICG) and the American Mycosis Research Group (MSG),and the American College of Infectious Diseases (IDSA) guidelines,295 patients with pulmonary aspergillosis (IPA) and high risk of invasive infections were divided into four groups: IPA group (42 cases),clinically diagnosed group (68 cases),suspected group (61 cases),and non-IPA group (124 cases). Their serum and BALF concentrations of GM were detected by enzyme-linked immunosorbent assay (ELISA).The clinically diagnosed and confirmed invasive pulmonary fungal infections (IPFI) were treated as golden standards (+). A GM value ≥ proposed threshold was deemed diagnostic test positive. Receiver operating characteristic (ROC) curves were drawn to determine the diagnostic efficiency of BALF GM assay for IPFI. The optimal cut-off point of BALF GM was determined using Youden index. RESULTS: BALF GM had an area under the curve (AUC) of 0.932 in diagnosing IPFI,with 87.5% sensitivity, 96.7% specificity, 87.5% positive predictive value,and 96.7% negative predictive value when the BALF GM value was set at 1.5 ng/mL as the optimal cut-off point. Higher BALF and serum GM values were found in the confirmed IPA group,followed by the clinical diagnosed group compared with the non-IPA group ( P<0.05). The threshold value was set at 0.5 ng/mL for serum GM and 1.5 ng/mL for bronchoalveolar lavage GM. Higher positive rates were found in the confirmed IPA group and the clinical diagnosed group compared with the non-IPA group ( P<0.05). Serum GM appeared to have higher false positives and false negative rates. CONCLUSION: BALF GM is a rapid and accurate indicator with high sensitivity and specificity for the early diagnosis of IPA.
Assuntos
Líquido da Lavagem Broncoalveolar/química , Aspergilose Pulmonar Invasiva/diagnóstico , Mananas/análise , Ensaio de Imunoadsorção Enzimática , Galactose/análogos & derivados , Humanos , Sensibilidade e EspecificidadeRESUMO
Cryptococcus is the major pathogen that causes fungal meningitis. In the People's Republic of China, especially in the Southwest area, cryptococcal meningitis (CM) in HIV-uninfected patients is more common than in HIV-infected patients. We compared clinical features and laboratory data pertaining to CM in patients with different immunological statuses. Demographic data, clinical manifestations, and laboratory data from inpatients in West China Hospital Sichuan University were collected from June 2009 to June 2014. Patients were grouped according to HIV status. Continuous variables were evaluated by Student t-test or Wilcoxon rank sum tests. Categorical variables were analyzed by χ2 test. Among 85 patients with CM were identified, 53 (62.4%) were HIV-uninfected patients. CM occurred more frequently in males in the HIV-infected group. Compared with HIV-infected patients, HIV-uninfected patients had more leukocytes in their blood and more leukocytes and protein in cerebrospinal fluid. More HIV-uninfected patients had increased cerebrospinal fluid (CSF)/serum albumin ratios, while intrathecal immunoglobulin G (IgG) synthesis was significantly increased. The rate of in-hospital mortality of HIV-infected CM patients was higher. Clinical signs are similar between HIV-uninfected and HIV-infected CM patients. Fewer leukocytes and protein was detected in the CSF and lower local synthesis of IgG in the central nervous system in HIV-infected patients, which reflects their diminished immune response. These characteristics should be noted in order to avoid misdiagnosis. Meningeal enhancement and intrathecal IgG synthesis in the HIV-uninfected group was significantly higher, that may be performance of aggressive inflammatory response and might contribute to a better outcome.
Assuntos
Cryptococcus/isolamento & purificação , Infecções por HIV/complicações , Meningite Criptocócica/patologia , Adolescente , Adulto , Idoso , China/epidemiologia , Feminino , Hospitais Universitários , Humanos , Masculino , Meningite Criptocócica/epidemiologia , Pessoa de Meia-Idade , Análise de Sobrevida , Adulto JovemRESUMO
BACKGROUND: Quorum Sensing (QS) systems influence biofilm formation, an important virulence factor related to the bacterial survival and antibiotic resistance. In Acinetobacter baumannii, biofilm formation depends on pili biosynthesis, structures assembled via the csuA/BABCDE chaperone-usher secretion system. QS signaling molecules are hypothesized to affect pili formation; however, the mechanism behind this remains unclear. This study aimed to demonstrate the possible role of QS signaling molecules in regulating pili formation and mediating the ability to form biofilms on abiotic surfaces. RESULTS: Real-time quantitative PCR analysis showed the expression of the csuA/BABCDE genes distinctly increased when co-cultured with C6-HSL (P < 0.05). Under the same experimental conditions, expression of BfmS and BfmR was significantly higher than the control strain (P < 0.05). A subsurface twitching assay showed a switch from a small to a large and structured clone that may result from enhanced twitching motility (P < 0.05). Transmission electron microscopy analysis of cells lifted from a MH broth co-cultured with C6-HSL showed more abundant pili-like structures than the control strain. We then tested the idea that the addition of a QS signal, and therefore induction of chaperone-usher secretion system genes, provides a greater benefit at higher biofilm densities. An assay for the total fluorescence intensity of the biofilm using Confocal Laser Scanning Microscopy revealed an obvious increase. CONCLUSION: Our study demonstrated that, increased transcription of the BfmS and BfmR genes, QS signaling molecules enhance the expression of the chaperone-usher secretion system, and this expression is required for twitching motility in A. baumannii. The concomitant pili expression and strain twitching allowed A. baumannii to attach easily to abiotic surfaces and form biofilms at an earlier timepoint.
Assuntos
Acinetobacter baumannii/efeitos dos fármacos , Acil-Butirolactonas/metabolismo , Biofilmes/efeitos dos fármacos , Fímbrias Bacterianas/efeitos dos fármacos , Locomoção/efeitos dos fármacos , Biogênese de Organelas , Acinetobacter baumannii/crescimento & desenvolvimento , Acinetobacter baumannii/metabolismo , Acinetobacter baumannii/fisiologia , Biofilmes/crescimento & desenvolvimento , Fímbrias Bacterianas/ultraestrutura , Perfilação da Expressão Gênica , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Previous reports on the molecular characteristics of clinical isolates of Cryptococcus species in China have focused on isolates from southeast China. To obtain a more detailed molecular epidemiology, a total of 92 cryptococcal isolates were collected from Sichuan province. A total of 24 isolates from 12 other provinces were collected for comparative study. Genotypes and mating types of 116 Cryptococcus isolates were determined. Among the 116 isolates, 43 isolates (19 isolates from Sichuan and 24 isolates outside of Sichuan) were analysed by multi-locus sequence typing (MLST). All 116 clinical isolates were mating type α. Most isolates (114/116) were molecular type VNI and the remaining two isolates were VGI and VGII respectively. MLST results revealed five sequence types (STs) of C. neoformans including two novel STs, with most isolates identified as ST5. The two C. gattii isolates identified in our study were ST44 and ST159. Based on our report and previous studies, there are 15 C. neoformans STs in China which can be divided into three subgroups. The C. gattii isolate from Sichuan could be a scattered subtype of VGII (ST44). Our findings demonstrated that C. neoformans isolates in Sichuan are genetically homogeneous, and ST5 is the epidemic clone of C. neoformans in China.
Assuntos
Criptococose/epidemiologia , Criptococose/microbiologia , Cryptococcus gattii/classificação , Cryptococcus gattii/genética , Cryptococcus neoformans/classificação , Cryptococcus neoformans/genética , China/epidemiologia , Cryptococcus gattii/isolamento & purificação , Cryptococcus neoformans/isolamento & purificação , Genes Fúngicos Tipo Acasalamento , Genótipo , Humanos , Tipagem de Sequências Multilocus , Técnicas de Tipagem Micológica , FilogeniaRESUMO
OBJECTIVE: To test the susceptibility of blood culture isolates to antibiotics in a sample of hospitals in Southwestern China in 2012. METHODS: Blood culture samples were taken and tested from the patients under the Whire Union surveillance in three tertiary hospitals in Southwestern China in 2012. We performed antimicrobial susceptibility analyses on the blood culture isolates using WHONET5. 5 and SPSS19.0. RESULTS: A total of 1745 isolates were identified: 877 Gram-positive (50.26%); 868 Gram-negative (49.74%). Coagulase-negative Staphylococcius (CNS, 541 strains, 31.00%), Escherichia coli (ECO, 379 strains, 21.72%), Klebsiella spp. (170 strains, 9.74%), Enterococcus spp. (143 strains, 8.20%) and Staphylococcus aureus (SAU, 128 strains, 7.34%) comprised the majority of isolates. Methicillin-resistant was found in 37.5% of S. aureus and 85.4% of CNS, respectively. Zero and 6.3% resistance to vancomycin were found for Enterococcus faecalis and E. faecium, respectively,compared with a 1.4% and 7.9% resistance to linezolid, respectively. Imipenem-resistance occurred in 0.7% E. coli, 3.6% Klebsiella spp., and 11.1% Enterobacter spp.. Nonfermenters were highly resistant to carbapenems, with an imipenem resistance rate of 25.0% in P. aeruginosa and 79.1% in Acinetobacter baumannii. Antimicrobial usage and critical-care units were identified as risk factors for MRSA and multi-drug-resistant A. baumannii infections. Patients undergoing tracheotomy/endotracheal intubation were likely to develop multi-drug-resistant A. baumannii infections than others. CONCLUSION: CNS, E. coli,Klebsiella spp., Enterococcus spp. and S. aureus were the predominant organisms in bloodstream infections in Southwestern China. Low methicillin-resistant rate was found in Staphylococcus aureus. But vancomycin-resistant rate was high in E. faecium. Imipenem-resistant rates varied in Enterobacteriaceae: higher in non-fermenting bacteria especially in Acinetobacter spp. Use of abtimicrobial drugs and invasive procedures can lead to development of antimicrobial drugs-resistance.
Assuntos
Antibacterianos , Bacteriemia/epidemiologia , Farmacorresistência Bacteriana , Acinetobacter baumannii , China/epidemiologia , Enterobacteriaceae , Enterococcus , Escherichia coli , Humanos , Resistência a Meticilina , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa , Staphylococcus aureusRESUMO
OBJECTIVE: To determine thepathogen characteristics of catheter-related bloodstream infection (CRBSI). METHODS: The clinical data of patients with CRBSI who were admitted in West China Hospital, Sichuan University during January 1, 2011 and October 15, 2014 were retrieved, along with findings of pathogen culture and drug susceptibility tests. RESULTS: Eighty-four strains of pathogens were isolated from 77 patients, which included 41 strains (48. 8%) of Gram-negative bacteria, 23 strains (27. 4%) of Gram-positive bacteria, and 20 strains of fungus (23. 8%). Enterobacteriaceae was predominant (29/41, 70.7%) in the Gram-negative bacteria,followed by non-fermenting bacteria (12/41, 29. 3%). Staphylococcus spp. was the main (16/23, 69. 6%) species of Gram- positive bacteria. Candida albicans led to 35. 0% (7/20) fungi infection. Resistance of Enterobacteriaceae to ceftriaxone was high (65. 5%, the highest), compared with its resistance to imipenem (3. 4%, the lowest). The non-fermentative bacterial had complete (100%) resistance to nitrofurantoin, and 16. 7% resistance (the lowest)to levofloxacin. Staphylococcus spp. had 81. 3% resistance (highest) to clindamycin, and zero resistance to vancomycin andlinezolid. Resistance to amphotericin and 5-flucytosine was not found in Candida spp. isolates. CONCLUSION: Prevention and control of CRBSI caused by Gram negative bacilli should be strengthened in the hospital. Clinical treatments should be guided by the in vitro drug susceptibility of pathogens.
Assuntos
Bacteriemia/microbiologia , Infecções Relacionadas a Cateter/microbiologia , Fungemia/microbiologia , Candida/isolamento & purificação , China , Farmacorresistência Bacteriana , Farmacorresistência Fúngica , Enterobacteriaceae/isolamento & purificação , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/isolamento & purificação , Humanos , Testes de Sensibilidade MicrobianaRESUMO
OBJECTIVE: Investigate the molecular epidemiological characteristics of bloodstream infections (BSI) due to Candida albicans (C. albicans) in the intensive care unit (ICU) of West China Hospital, Sichuan University. METHODS: C. albicans isolates recovered from blood cultures in West China Hospital, Sichuan University between 2009 and 2011 were collected. Multilocus sequence typing (MLST) was performed to assess the genetic relationships among BSI isolates of C. albicans collected from the ICU. RESULTS: 135 BSI isolates of Candida species were obtained from 2009 to 2011. C. albicans was the leading pathogen (51 isolates, 37.8%). 17 C. albicans BSI isolates from 15 patients of ICU were analyzed by MLST. Among the 17 isolates, 15 were recovered from peripheral blood and 2 from central venous catheters (CVC) (Peripheral blood and CVC were sent for culture and both had positive results for 2 patients). The 17 isolates yielded 15 unique sequence types (STs) by MLST. While 14 STs were each derived from a single isolate, 1 STs were shared by 3 isolates. 5 (29.4%) isolates were clustered within Group 46, 2 (11.8%) isolates were clustered within Group 47, and 10 isolates (58.8%) typed as singletons. The strains (Calb-36 and Calb-40) recovered from one blood sample and one CVC from one patient were indistinguishable by MLST, while two distinct strains were found in one blood sample and one CVC from another patient. CONCLUSION: C. albicans was the most frequently isolated species of candidemia in West China Hospital. Predominant strains of C. albicans caused BSI in the ICU belonged to Group 46 and Group 47. There was not yet an outbreak of BSI caused by C. albicans, but catheter-related candidemia was confirmed by our research.
Assuntos
Candida albicans/isolamento & purificação , Candidemia/microbiologia , Infecções Relacionadas a Cateter/microbiologia , Unidades de Terapia Intensiva , Tipagem de Sequências Multilocus , Candida albicans/classificação , Candida albicans/genética , Candidemia/epidemiologia , Candidíase/epidemiologia , China/epidemiologia , Infecção Hospitalar/microbiologia , Humanos , Epidemiologia Molecular , Técnicas de Tipagem Micológica , FilogeniaRESUMO
OBJECTIVE: To study the influence of modified Shegan Mahuang Decoction (SGMH) on cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-10 and IL-13 in children suffered from cough and variant asthma (C&VA). METHODS: One hundred and fifty-four children with C&VA were randomly assigned to two groups: 79 in the treatment group were medicated orally with SGMH one dose per day taking in twice; 75 in the control group were medicated with Montelukast once a day in dose of 4 mg for children aged from 2 to 5 years and 5 mg for those from 6 to 14 years, the medication for all was given 4 weeks. Serum contents of cytokines, including TNF-alpha, IL-10 and IL-13, in patients were measured before and after treatment. Besides, serum contents of these cytokines in 45 healthy children were measured for control. RESULTS: Serum levels of TNF-alpha and IL-3 in the treatment group were 2510 +/- 1500 ng/L and 60.76 +/- 23.67 ng/L, and in the control group, 2890 +/- 1410 ng/L and 61.56 +/- 20.37 ng/L, respectively, all were significantly higher than those of healthy (709 +/- 280 ng/L and 39.49 +/- 3.09 ng/L, P < 0.01); but level of IL-10 was significant lower in the two patient groups than that in control (1546 +/- 1434 ng/L and 1823 +/- 1314 ng/L vs 7123 +/- q2641 ng/L, P < 0.01). After treatment, the levels of TNF-alpha and IL-13 decreased and IL-10 increased significantly in the treatment group, and showed significant different to those in the control group respectively (960 +/- 420 ng/L, 43.67 +/- 12.37 ng/L and 6834 +/- 2216 ng/L vs 2610 +/- 1220 ng/L, 50.56 +/-19.56 ng/L and 2529 +/- 1223 ng/L, P < 0.01). Clinical efficacy between groups also showed that the total effective rate in the treatment group was significantly better (86.07% vs. 42.67%, P < 0.01). CONCLUSION: SGMH can regulate the serum levels of TNF-alpha, IL-10 and IL-13, and shows evident clinical effect in treating children's C&VA.
Assuntos
Asma/sangue , Tosse/sangue , Medicamentos de Ervas Chinesas/uso terapêutico , Fitoterapia , Adolescente , Asma/tratamento farmacológico , Criança , Pré-Escolar , Tosse/tratamento farmacológico , Ephedra sinica , Feminino , Humanos , Lactente , Interleucina-10/sangue , Interleucina-13/sangue , Interleucina-3/sangue , Masculino , Fator de Necrose Tumoral alfa/sangueRESUMO
OBJECTIVE: To analyze the etiology of infections in the wounded victims of Wenchuan Earthquake. METHODS: 2135 smears of secretion were made from 1823 hospitalized wounded victims of Wenchuan Earthquake to detect the pathogens. Specimens were delivered to be cultured. The bacteria thus obtained were identified. Drug sensitivity test was conducted. RESULTS: 2002 specimens, 1243 specimens of secretion (62.1%), 600 blood specimens (30.0%), 102 specimens of pus or secretion of respiratory tract (5.1%), 45 specimens from catheter (2.2%), and 12 urine specimens (0.5%). Pathogens were found in 725 cases. The top five pathogenic bacteria isolated within 1 month after the quake were Acinetobacter baumannii (16.7%), Staphylococcus haemolyticus (16.7%), Escherichia coli (12.5%), Klebsiella pneumoniae (12.5%), and Candida tropicalis (8.3%), quite different from the pathogen spectrum of the common in-patients within one month before the quake: Escherichia coli (18.2%), Staphylococcus aureus (11.4%), Candida glabrata (11.4%), Klebsiella pneumoniae (11.4%) and Staphylococcus epidermidis (9.1%). The isolation rate of methicillin resistant Staphylococcus aureus after the earthquake was significantly lower, and the isolation rates of extended spectrum beta-lactamase-producing Escherichia coli, Klebsiella pneumoniae, and Klebsiella oxytoca were all significantly higher than those from the common surgical patients before the quake (all P < 0.05). There were not significant differences in the isolation rates of multi-drug resistant Pseudomonas alcaligenes and Acinetobacter baumannii before and after the quake. CONCLUSION: Infection is frequent after natural disasters. It is necessary to summarize the changes of spectrum of pathogens and drug-resistant spectrum.
Assuntos
Infecções Bacterianas/etiologia , Desastres , Terremotos , Ferimentos e Lesões/microbiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , China , Farmacorresistência Bacteriana Múltipla , Feminino , Humanos , Lactente , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Adulto JovemRESUMO
BACKGROUND: The pathogenesis of multiple sclerosis (MS) is mediated primarily by T cells, but most studies of MS and its animal model, experimental autoimmune encephalomyelitis (EAE), have focused on CD4 T cells. The aims of the current study were to determine the pathological interrelationship between CD4 and CD8 autoreactive T cells in MS/EAE. METHODS: Female C57BL/6 mice (nâ=â20) were induced by myelin oligodendrocyte glycoprotein (MOG)35-55 peptide. At 14 days after immunization, T cells were isolated from the spleen and purified as CD4 and CD8 T cells by using CD4 and CD8 isolation kits, and then the purity was determined by flow cytometric analysis. These cells were stimulated by MOG35-55 peptide and applied to proliferation assays. The interferon-gamma (IFN-γ) and interleukin (IL)-4 secretion of supernatant of cultured CD4 and CD8 T cells were measured by enzyme-linked immunosorbent assays (ELISA). For adoptive transfer, recipient mice were injected with MOG35-55-specific CD8 or CD4 T cells. EAE clinical course was measured by EAE score at 0-5 scale and spinal cord was examined by staining with hematoxylin and eosin and Luxol fast blue staining. RESULTS: CD8CD3 and CD4CD3 cells were 86% and 94% pure of total CD3 cells after CD8/CD4 bead enrichment, respectively. These cells were stimulated by MOG35-55 peptide and applied to proliferation assays. Although the CD8 T cells had a generally lower response to MOG35-55 than CD4 T cells, the response of CD8 T cells was not always dependent on CD4. CD8 T cell secreted less IFN-γ and IL-4 compared with CD4 T cells. EAE was induced in wildtype B6 naïve mice by adoptive transfer of MOG35-55-specific T cells from B6 active-induced EAE (aEAE) mice. A similar EAE score and slight inflammation and demyelination were found in naive B6 mice after transferring of CD8 T cells from immunized B6 mice compared with transfer of CD4 T cells. CONCLUSION: Our data suggest that CD8 autoreactive T cells in EAE have a lower encephalitogenic function but are unique and independent on pathogenic of EAE rather than their CD4 counterparts.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Encefalomielite Autoimune Experimental/imunologia , Glicoproteína Mielina-Oligodendrócito/imunologia , Transferência Adotiva , Animais , Linfócitos T CD4-Positivos/imunologia , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/etiologia , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Fragmentos de Peptídeos/imunologiaRESUMO
The venom of Crotalus durissus terrificus is reported to have analgesic activity and the administration of Crotoxin (Cro) to cancer patients is reported to reduce the consumption of analgesics. This study investigated the analgesia induced by Cro and the effects of atropine and naloxone on the antinociceptive activity of Cro in mice and rats. The results showed that Cro at 66.5, 44.3 and 29.5microg/kg (ip) exhibited a dose-dependent analgesic action in mice using the hotplate and acetic acid writhing tests. Cro at 44.3microg/kg (ip) had significant analgesic action in the rat tail-flick test. In the mouse acetic acid-writhing test, intracerebral ventricular administration of Cro 0.3microg/kg produced marked analgesic effects. Microinjection of Cro (0.15microg/kg) into the periaqueductal gray area also elicited a robust analgesic action in rat hotplate test. Atropine at 0.5mg/kg (im) or 10mg/kg (ip) or naloxone at 3mg/kg (ip) failed to block the analgesic effects of Cro. These results suggest that Cro has analgesic effects mediated by an action on the central nervous system. The muscarinic and opioid receptors are not involved in the antinociceptive effects of Cro.
Assuntos
Acetilcolina/antagonistas & inibidores , Analgésicos Opioides/antagonistas & inibidores , Analgésicos/farmacologia , Venenos de Crotalídeos/química , Crotalus , Crotoxina/farmacologia , Analgésicos/administração & dosagem , Analgésicos/isolamento & purificação , Animais , Atropina/farmacologia , Crotoxina/administração & dosagem , Crotoxina/isolamento & purificação , Relação Dose-Resposta a Droga , Interações Medicamentosas , Feminino , Injeções Intraventriculares , Masculino , Camundongos , Camundongos Endogâmicos , Antagonistas Muscarínicos/farmacologia , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Medição da Dor/efeitos dos fármacos , Limiar da Dor/efeitos dos fármacosRESUMO
AIM: To seek the X associated protein (XAP) with the constructed bait vector pAS2-1X from normal human liver cDNA library. METHODS: The X region of the HBV gene was amplied by PCR and cloned into the eukaryotic expression vector pAS2-1. The reconstituted plasmid pAS2-1X was transformed into the yeast cells and the expression of X protein (pX) was confirmed by Western blot analysis. Yeast cells were cotransformed with pAS2-1X and the normal human liver cDNA library and were grown in selective SC/-trp-leu-his-ade medium, the second screen was performed with the LacZ report gene. Furthermore, segregation analysis and mating experiment were performed to eliminate the false positive and the true positive clones were selected for PCR and sequencing. RESULTS: Reconstituted plasmid pAS2-1X including the anticipated fragment of X gene was proved by auto-sequencing assay. Western blot analysis showed that reconstituted plasmid pAS2-1X expressed BD:X fusion protein in yeast cells. Of 5 x 10(6) transformed colonies screened,65 grew in the selective SC/-trp-leu-his-ade medium, 5 scored positive for beta-gal activity, and only 2 remaining clones passed through the segregation analysis and mating experiment. Sequence analysis identified that two clones contained similar cDNA fragment:GAACTTGCG. CONCLUSION: The short peptide(glutacid-leucine-alanine)is a possible required site for XAP binding to pX. Normal human liver cDNA library has difficulties in expressing the integrated XAP on yeast cells.
Assuntos
Fígado/fisiologia , Transativadores/genética , Transativadores/metabolismo , Sequência de Bases , Western Blotting , Biblioteca Gênica , Testes Genéticos , Humanos , Fígado/química , Dados de Sequência Molecular , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Plasmídeos , Saccharomyces cerevisiae , Técnicas do Sistema de Duplo-Híbrido , Proteínas Virais Reguladoras e AcessóriasRESUMO
AIM: To systematically review the evidence for the effectiveness of fast-track program vs traditional care in laparoscopic or open surgery for gastric cancer. METHODS: PubMed, Embase and the Cochrane library databases were electronically searched for published studies between January 1995 and April 2013, and only randomized trials were included. The references of relevant studies were manually searched for further studies that may have been missed. Search terms included "gastric cancer", "fast track" and "enhanced recovery". Five outcome variables were considered most suitable for analysis: postoperative hospital stay, medical cost, duration to first flatus, C-reactive protein (CRP) level and complications. Postoperative hospital stay was calculated from the date of operation to the date of discharge. Fixed effects model was used for meta-analysis. RESULTS: Compared with traditional care, fast-track program could significantly decrease the postoperative hospital stay [weighted mean difference (WMD) = -1.19, 95%CI: -1.79--0.60, P = 0.0001, fixed model], duration to first flatus (WMD = -6.82, 95%CI: -11.51--2.13, P = 0.004), medical costs (WMD = -2590, 95%CI: -4054--1126, P = 0.001), and the level of CRP (WMD = -17.78, 95%CI: -32.22--3.35, P = 0.0001) in laparoscopic surgery for gastric cancer. In open surgery for gastric cancer, fast-track program could also significantly decrease the postoperative hospital stay (WMD = -1.99, 95%CI: -2.09--1.89, P = 0.0001), duration to first flatus (WMD = -12.0, 95%CI: -18.89--5.11, P = 0.001), medical cost (WMD = -3674, 95%CI: -5025--2323, P = 0.0001), and the level of CRP (WMD = -27.34, 95%CI: -35.42--19.26, P = 0.0001). Furthermore, fast-track program did not significantly increase the incidence of complication (RR = 1.39, 95%CI: 0.77-2.51, P = 0.27, for laparoscopic surgery; and RR = 1.52, 95%CI: 0.90-2.56, P = 0.12, for open surgery). CONCLUSION: Our overall results suggested that compared with traditional care, fast-track program could result in shorter postoperative hospital stay, less medical costs, and lower level of CRP, with no more complications occurring in both laparoscopic and open surgery for gastric cancer.
Assuntos
Gastrectomia/métodos , Laparoscopia , Cuidados Pós-Operatórios/métodos , Neoplasias Gástricas/cirurgia , Redução de Custos , Análise Custo-Benefício , Procedimentos Clínicos , Gastrectomia/economia , Custos Hospitalares , Humanos , Laparoscopia/economia , Tempo de Internação/economia , Cuidados Pós-Operatórios/economia , Avaliação de Programas e Projetos de Saúde , Recuperação de Função Fisiológica , Neoplasias Gástricas/economia , Fatores de Tempo , Resultado do TratamentoRESUMO
The successful synthesis of the highly complex model compound (2) of the CEFGH ring system of schindilactoneâ A (1) is described. Several synthetic methodologies were developed and applied to achieve this goal, including ring-closing metathesis (RCM) and Co-thiourea-catalyzed Pauson-Khand reactions. Furthermore, two independent approaches were developed for the construction of the GH ring of model compound 2, the key steps of which included Pd-thiourea-catalyzed carbonylative annulation, methylation, and sequential RCM/oxa-Michael-addition reactions. The chemistry developed herein has provided a greater understanding of the synthesis of schindilactoneâ A (1) and its analogous compounds of the same family.
Assuntos
Triterpenos/síntese química , Catálise , Cristalografia por Raios X , Ciclização , Lactonas/química , Metilação , Conformação Molecular , Paládio/química , Estereoisomerismo , Tioureia/química , Triterpenos/químicaRESUMO
The final phase for the total synthesis of (±)-schindilactoneâ A (1) is described herein. Two independent synthetic approaches were developed that featured Pd-thiourea-catalyzed cascade carbonylative annulation reactions to construct intermediate 3 and a RCM reaction to make intermediate 4. Other important steps that enabled the completion of the synthesis included: 1)â A Ag-mediated ring-expansion reaction to form vinyl bromide 17 from dibromocyclopropane 30; 2)â a Pd-catalyzed coupling reaction of vinyl bromide 17 with a copper enolate to synthesize ketoester 16; 3)â a RCM reaction to generate oxabicyclononenol 10 from diene 11; 4)â a cyclopentenone fragment in substrate 8 was constructed through a Co-thiourea-catalyzed Pauson-Khand reaction (PKR); 5)â a Dieckmann-type condensation to successfully form the Aâ ring of schindilactoneâ A (1). The chemistry developed for the total synthesis of schindilactoneâ A (1) will shed light on the synthesis of other family members of schindilactoneâ A.
Assuntos
Triterpenos/síntese química , Catálise , Cristalografia por Raios X , Reação de Cicloadição , Conformação Molecular , Paládio/química , Estereoisomerismo , Tioureia/química , Triterpenos/químicaRESUMO
BACKGROUND & OBJECTIVE: Mucoepidermoid carcinoma is the most common malignant tumor of the salivary, but mucoepidermoid carcinoma of the parotid gland has seldom been reported. This study was to summarize the clinicopathologic prognostic factors of mucoepidermoid carcinoma of the parotid gland. METHODS: Clinical data of 116 patients with mucoepidermoid carcinoma of the parotid gland, treated in Cancer Center of Sun Yat-sen University from May 1980 to Dec. 2000, were analyzed with Cox univariate and multivariate models. RESULTS: The overall 5-, 10-, and 15-year survival rates of the 116 patients were 75.64%, 64.55%, and 60.39%, respectively. Univariate survival analysis showed that 12 factors were prognostic factors of mucoepidermoid carcinoma of the parotid gland, such as age, alcohol drinking, T stage, and so on. Multivariate analysis showed that T stage (P = 0.006, OR > 1), pathologic grade (P < 0.001, OR > 1), and distant metastasis (P < 0.001, OR > 1) were independent prognostic factors of mucoepidermoid carcinoma of the parotid gland. CONCLUSION: T stage, pathologic grade and distant metastasis are independent prognostic factors of mucoepidermoid carcinoma of the parotid gland.