Objective: Reveal the global expression profile of serum exosomal proteins of Crouzon syndrome patients. Methods: We isolated microvesicles from serum of Crouzon children with a C342Y mutation in FGFR2 by ultracentrifugation, which were further characterized by electron microscopy and immunoblotting. The protein profiling in normal subjects and Crouzon patients was systematically compared by iTRAQ proteomic analysis. Results: The result demonstrated that microvesicles were between 30—100 nm in diameter, round shape with cup-like concavity and expressed exosomal marker tumor susceptibility gene (TSG) 101 and flotillin (Flot) 1. We identified a total number of 62 proteins, among which 22 proteins overlap with ExoCarta database and were different between the Crouzon patient and the normal subject. The Ingenuity Pathway Analysis showed that the functions of these proteins are mostly involved in Developmental Disorder, Hereditary Disorder, Skeletal and Muscular Disorders, which are all related to the clinical manifestations of Crouzon syndrome. In addition, the proteins were focused on the network of "Organismal Injury and Abnormalities, Hematological System Development and Function, Cell-To-Cell Signaling and Interaction" . The central protein FN1 was presented as the key protein in the network. Conclusions Our data demonstrated that serum exosomes harbor informative proteins and FN1 was selected as a potential candidate for its role in promoting osteoblast adhesion, proliferation and mineralization.

Objective To evaluate the effect of up-converting phosphor technology(UPT) in detection of plague antigen-antibody by receiver operating characteristic curve (ROC) method,and to provide a scientific basis for field application of UPT rapid detection technology in plague prevention and control.Methods Two hundred and twenty four serum samples were collected from Marmots and ground squirrels in the plague foci,Yersinia pestis antibody was detected by UPT,ELISA,Colloidal-gold Strips and IHA,respectively; 108 organs and bone marrow samples were collected,and Yersinia pestis antigens were detected by UPT,ELISA,PCR and RIHA,respectively.IHA was used as the gold standard for antibody test results,RIHA,PCR + Colloidal-gold Strips,PCR + ELISA were used as the gold standard for antigen test results.The results were evaluated using ROC method.Results Antibodies detection:the AUCs of UPT,ELISA and Colloidal-gold Strips were greater than 0.5.The difference between UPT and other methods was not statistically significant (z =1.204,P ＞ 0.05).Antigen detection:the AUCs of UPT,ELISA,Colloidal-gold Strips and PCR were greater than 0.5.There was no statistical difference between UPT and other methods(z =0.866,P ＞ 0.05).Conclusions UPT as a new technology works well in the detection of plague antigen-antibody.The technology is simple,fast,accurate,and suitable for on-site monitoring of plague,emergency treatment of sudden plague,and suitable for promotion.

OBJECTIVETo study the relationship between morphological characteristics of growth cycle for Epimedium acuminatium and accumulation level of secondary metabolites.

METHODAfter making habitat clear, methods of morphology observation, specimen preparation, organism charting were applied to record morphological characteristics of E. acuminatium at different stages. HPLC was employed to detect icariin content of different parts of the plant at the stage of vegetative growth and sexual reproduction.

RESULTThe growth cycle of E. acuminatium was divided into vegetative growth stage and sexual and asexual reproduction coexistence stage. Seven to eight years were needed for the plants flowering and seeding of E. acuminatimcan in open land of forest edge, but within good nutrition conditions, growth cycle is shortened into three to four years. The difference of icariin content in two growth stages was not significant.

CONCLUSIONMorphological change of lower and foliar organ can be utilized to estimate growth age. But after applying sexual propagation for both root and stem of grown plants, it's been difficult to determine their growth year. If individual biomass or yield is economical, plants can be harvested in both vegetative growth and sexual reproduction stages.

Biomass ; Chromatography, High Pressure Liquid ; methods ; Epimedium ; growth & development ; metabolism ; Flavonoids ; metabolism ; Plant Roots ; growth & development ; metabolism ; Reproduction ; Seedlings ; metabolism

Ischemic stroke leads to high potentiality of mortality and disability.The current treatment for ischemic stroke is mainly focused on intravenous thrombolytic therapy.However,ischemia/reperfusion induces neuronal damage,which significantly influences the outcome of patients with ischemic stroke,and the exact mechanism implicated in ischemia/reperfusion injury remains unclear,although evidence shows that oxidative stress is likely to be involved.Betulinic acid is mainly known for its anti-tumor and anti-inflammatory activities.Our previous study showed that betulinic acid could decrease the reactive oxygen species (ROS) production by regulating the expression of NADPH oxidase.Thus,we hypothesized that betulinic acid may protect against brain ischemic injury in the animal model of stroke.Focal cerebral ischemia was achieved by using the standard intraluminal occlusion method and reperfusion enabled after 2 h ischemia.Neurological deficits were scored.Infarct size was determined with 2,3,5-triphenyltetrazolium chloride monohydrate (TTC) staining and the mRNA expression of NADPH oxidase 4 (NOX4) was determined by RT-PCR in infarct tissue.ROS generation and apoptosis in ischemic tissue were analyzed by measuring the oxidative conversion of cell permeable 2',7'-dichloro-fluorescein diacetate (DCF-DA) to fluorescent dichlorofluorescein (DCF) in fluorescence microplate reader and TUNEL assay,respectively.In Kunming mice,2 h of middle cerebral artery (MCA) occlusion followed by 24 or 72 h of reperfusion led to an enhanced NOX4 expression in the ischemic hemisphere.This was associated with elevated levels of ROS generation and neuronal apoptosis.Pre-treatment with betulinic acid (50 mg/kg/day for 7 days via gavage) prior to MCA occlusion prevented the ischemia/reperfusion-induced up-regulation of NOX4 and ROS production.In addition,treatment with betulinic acid could markedly blunt the ischemia/reperfusion-induced neuronal apoptosis.Finally,betulinic acid reduced infarct volume and ameliorated the neurological deficit in this stroke mouse model.Our results suggest that betulinic acid protects against cerebral ischemia/reperfusion injury in mice and the down-regulation of NOX4 may represent a mechanism contributing to this effect.

OBJECTIVE: To study the disability identification for cases with clinical diagnosis of diffuse axonal injury (DAI) due to traffic accidents, and to explore the possible effects of DAI on identification results. METHODS: Five hundred and fifty-six cases of cerebral injury due to traffic accidents were collected, including 467 cases diagnosed with cerebral contusion or laceration and 89 cases diagnosed with DAI. The identification results of different groups with diagnosis of DAI diagnosis, diagnosis of DAI with cerebral contusion (laceration), and diagnosis of cerebral contusion or laceration without DAI were compared and statistically analyzed, based on the results of CT and MRI re-review. RESULTS: The disability identification levels in DAI group (20 cases), DAI group (69 cases) with cerebral contusion (laceration) and DAI group (467 cases) not complicated by cerebral contusion (laceration) were 7.72 +/- 1.09, 7.78 +/- 1.11, and 8.86 +/- 0.66, respectively. The disability levels of the two groups diagnosed with DAI were higher than those of the group without DAI diagnosis (P < 0.05). CONCLUSION Patients with DAI diagnosis might have more severe cerebral injury. In the identification process, one should pay attention to the possible missed diagnosis and misdiagnosis, and meanwhile avoid relying on those evidences provided only by CT and MRI.
Accidents, Traffic ; Brain Injuries/diagnosis* ; Diagnostic Errors ; Diffuse Axonal Injury/etiology* ; Disability Evaluation ; Forensic Pathology ; Humans ; Magnetic Resonance Imaging ; Resin Cements ; Tomography, X-Ray Computed

The aim of the study is to evaluate the utility value of species identification of Triculinae based on COI gene.Triculinae-like samples were collected from different places in Fujian Province.After classification based on morphology characters,genomic DNA was extracted from collected samples.Then the segments of COI gene were amplified and sequenced for taxonomy annotation and phylogenetic analysis.Eleven samples were annotated as Gammatricula (identities:88.96％-97.82％),the other two were annotated as Tricula wumingensis (identity:87.08％) and Neotricula apart (identity:88.55％),respectively.In most cases (12 out of 13),there was a difference between results based on different classification methods on a genus level.The alignment of COI gene segment is sufficient for preliminary identification of Triclulinae at family level.However,it is need further study in species identification of Triclulinae at genus level.

OBJECTIVETo explore the effects of p,p'-DDE and beta-BHC on the apoptosis of Sertoli cells in vitro via activation of Caspase.

METHODSSertoli cells were treated in vitro for 24 hours with a serial concentrations of p,p'-DDE (10, 30 and 50 micromol/L), beta-BHC (10, 30 and 50 micromol/L) and p,p'-DDE + beta-BHC (10, 30 and 50 micromol/L). The inhibitory group was first treated with 100 micromol/L Caspase-3 inhibitor Ac-DEVD-CHO treating for 2 hours before 50 micromol/L p, p'-DDE + 50 micromol/L beta-BHC 24 hours-treatment. The vitality of Sertoli cells was determined by MTT and the apoptosis rate was measured by AO/EB double fluorescence staining. The expressions of Caspase-3, Caspase-8 and Caspase-9 were determined by RT-PCR.

RESULTSAverage optical density (A) values were 0.498 +/- 0.039, 0.481 +/- 0.065, 0.397 +/- 0.032 and 0.286 +/- 0.049 in p,p'-DDE groups (10, 30, 50 and 70 micromol/L), and 0.518 +/- 0.103, 0.490 +/- 0.060, 0.454 +/- 0.054 and 0.302 +/- 0.030 in beta-BHC groups (10, 30, 50 and 70 micromol/L). In the mixture-treated groups (10, 30 and 50 micromol/L), the average A values were 0.483 +/- 0.048, 0.473 +/- 0.058 and 0.337 +/- 0.052. Compared with the solvent control group (0.527 +/- 0.022) , 50 micromol/L group of p, p'-DDE, beta-BHC or their mixture caused a significant decrease of Sertoli cell viability (t values were 4.599, 2.716, 6.537 respectively, P < 0.05). AO/EB double fluorescence staining analysis showed that apoptosis rates of Sertoli cells were significantly increased with all treated groups. The expressions of Caspase-3, Caspase-8 and Caspase-9 were upregulated as the concentrations of p,p'-DDE, beta-BHC and their mixture were increased.

CONCLUSIONp,p'-DDE, beta-BHC and their mixture could induce the apoptosis of Sertoli cells in vitro which was associated with activation of Caspase-3 mediated by cleavage of Caspase-8 and Caspase-9.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cells, Cultured ; Dichloroethylenes ; toxicity ; Lindane ; toxicity ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; drug effects ; metabolism

OBJECTIVETo study the action mechanism of p,p'-DDE and/or beta-BHC on JNK and MAPK signal transduction pathways in rat Sertoli cells in vitro.

METHODSWe cultured the Sertoli cells isolated from rat testicular tissues for 2 days in vitro, divided them into a control group incubated with DMSO and 3 case groups exposed to p,p'-DDE and / or beta-BHC at the final concentration of 10, 30, 50 micromol/L for 24 hours, and then detected the expression levels of JNK and c-jun mRNA by two-step RT-PCR.

RESULTSTwenty-four hours after p,p'-DDE treatment, the grayscale values of JNK mRNA were 0.068 +/- 0.001, 0.164 +/- 0.002, 0.207 +/- 0.006 and 0.499 +/- 0.017, and those of c-jun mRNA were 0.122 +/- 0.002, 0.157 +/- 0.006, 0.218 +/- 0.007 and 0.289 +/- 0.004 respectively in the DMSO control and the 10, 30 and 50 micromol/L groups. The expressions of JNK and c-jun mRNA were elevated with increased concentration of p,p'-DDE, with significant differences between the control and the case groups (P < 0.05), and they were also significantly upregulated in the beta-BHC and p,p'-DDE + beta-BHC groups in a dose-dependent manner. The grayscale values of JNK mRNA in the p,p'-DDE, beta-BHC and p,p'-DDE + beta-BHC groups at the concentration of 10 micromol/L were 0.164 +/- 0.002, 0.149 +/- 0.003 and 0.178 +/- 0.004, and those of c-jun mRNA were 0.157 +/- 0.006, 0.131 +/- 0.004 and 0.172 +/- 0.002, respectively, both significantly higher in the combination group than in the former two (P < 0.05). And the same was the case with the 30 and 50 micromol/L concentrations.

CONCLUSIONBoth p,p'-DDE and beta-BHC can enhance the expressions of JNK and c-jun in Sertoli cells, and their combination can produce even more obvious effect.

Animals ; Cells, Cultured ; Dichlorodiphenyl Dichloroethylene ; chemistry ; toxicity ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Lindane ; chemistry ; toxicity ; MAP Kinase Signaling System ; drug effects ; Male ; Proto-Oncogene Proteins c-jun ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sertoli Cells ; cytology ; drug effects ; metabolism

Objective To investigate the prognosis of adenoid cystic carcinoma (ACC) in salivary gland and its influencing factors.Methods Clinical and following-up data of 76 patients with ACC in salivary glands were reviewed. Major gland tumors represented 35.5％ whereas minor gland tumors comprised 64.5％ of the cohort,with 8 cases ( 10.5％ ) in stage I,23 (30.3％ ) in stage Ⅱ,18 (23.7％) in stage Ⅲ and 27 (35.5％) in stage Ⅳ. Survival rates were calculated by Kaplan-Merier method.Cumulative survival curves were evaluated using the Log-rank test. Multivariate analysis was performed by Cox proportional hazard model.Results The regional recurrence rate was 28.9％ and distant metastasis rate was 21.1％.The overall 5-year survival rate,tumor-free survival rate and tumor-related survival rate were 73.7％,61.8％ and 74.9％ respectively.The overall 10-year survival rate,tumor-free survival rate and tumor-related survival rate were 48.2％,39.8％ and 56.2％ respectively.Univariate survival analysis showed pathological type,clinical stage and perineural invasion were relevant to the prognosis of ACC and multivariate analysis showed they were the independent prognostic factors of ACC in salivary gland.Conclusions Clinical stage,pathological type and perineural invasion were the independent prognostic factors for adenoid cystic carcinoma in salivary gland. Surgery was the first choice for the treatment of adenoid cystic carcinoma in salivary gland,and postoperative radiotherapy may prolong the tumor-free survival time of patients in stage Ⅲ and Ⅳ.

Animals ; China ; epidemiology ; Genotype ; Geography ; Humans ; Marmota ; microbiology ; Plague ; epidemiology ; microbiology ; Yersinia pestis ; classification ; genetics ; isolation & purification