OBJECTIVETo establish and optimize the proteomic analysis of protoscoleces-specific antigens from Echinococcus granulosus. To provide a foundation for identifying specific antigens in the soluble proteins of E. granulosus protoscoleces for further research.

METHODSBrood capsules were collected aseptically from fertile E. granulosus cysts from the livers of an infected patient. The fertile E. granulosus cysts were fractured, and protoscoleces were collected by centrifugation. The soluble proteins of protoscoleces were acquired using the 2D Quant kit according to the manufacturer's instructions. We employed two-dimensional electrophoresis (2-DE) combined with immunoblot assay (Western blot) to analyze the soluble components of E. granulosus protoscoleces antigens. The 2-DE and immunoblot maps obtained were analyzed with PDQuest 8.0 image analysis software.

RESULTSAbout 233 soluble protein spots were identified with Coomassie-stained gels. Most of the proteins had a molecular weight of 16,000 Da to 117,000 Da, and an isoelectric point value of 3.0 to 10.0. 2-DE immunoblot was conducted and 57 specific antigen spots were observed, among which 23 spots were identified.

CONCLUSION2-DE combined with Western blot is the key to successful proteomic analysis and presents a new possibility for searching the specific E. granulosus protoscoleces antigens.

Animals ; Antigens, Helminth ; chemistry ; metabolism ; Blotting, Western ; methods ; Echinococcus granulosus ; classification ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Gene Expression Regulation ; Helminth Proteins ; isolation & purification ; Proteomics ; methods

Amiodarone ; adverse effects ; Female ; Humans ; Middle Aged ; Muscular Diseases ; chemically induced

AIM To study the mRNA expression of histamine H_1 receptor in hepatocarcinoma of rats. METHODS Dimethylamino-azobenzene (DAB) was used to induce hepatocarcinoma in rats. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was adopted to analyse the relative expression of histamine H_1 receptor. And the base sequence of its PCR product was detected. RESULTS The relative mRNA expression of histamine H_1 receptor was significantly decreased in hepatic carcinoma tissue, compared with that part far from cancer and control group (P

?AIM:To investigate the measurement of central anterior chamber depth ( ACD ) in patients with acute primary angle - closure glaucoma ( APACG ) with A - scan ultrasound, Pentacam and ultrasonic biological microscope ( UBM) .?METHODS: Thirty-five patients (35 eyes) with APACG were selected, of whom central ACD were measured with A-scan ultrasound, Pentacam and UBM.?RESULTS: The measurement values of ACD with A-scan ultrasound, UBM and Pentacam were 1. 5633±0. 2089, 1. 5783 ± 0. 2067, 1. 6275 ± 0. 2296mm, which was equal variance tested by the homogeneity of variance, and was significant different by multiple comparision (F=4. 074, P=0. 026). The difference of ACD between the two groups of A-scan ultrasound and UBM, A-scan ultrasound and Pentacam, UBM and Pentacam were statistically significant ( P = 0. 032, 0. 023, 0. 012 ). Altman- Bland analysis showed that the three methods were not consistent with each other.?CONCLUSION: The ACD value of the APACG with the three methods is the largest using Pentacam, followed by UBM and A - scan ultrasound. In clinical the three methods with different advantages can complement each other, but cannot be replaced. In order to obtain more accurate results, we should combine the advantage and make comprehensive analysis.

Background Some researching results of sodium hyaluronate eye drops treating dry eye have been reported home and abroad.To evaluate these results using the method of evidence-based medicine (EBM) has an important clinical significance.Objective This secondary study was to systematically evaluate the efficacy of sodium hyaluronate eye drops for the treatment of dry eye.Methods Meta analysis was adopted in this study.The electronic bibliographic databases were searched according to the prescribed strategy,including Medline,PubMed,VIP,CBM,CNKI,Wanfang Data using the method of Cochrane systematic review.The literature published time was limited from the commencement of each database to December,2011.Randomized controlled trials (RCTs) of sodium hyaluronate eye drops treating dry eye were included and the methodological quality of the RCTs was assessed by two evaluators using Jadad measuring scale,and homogeneous evaluation by Meta-analysis was performed.Statistical analysis of clinical data was performed using RevMan 5.0 software provided by the Cochrane Collaboration.With odds ratio (OR) value as the effect index,the randomized effect model was used to analyze the clinical heterogeneity.Results A total of 4 RCTs involving 686 patients were included.Jadad scale evaluation showed 7 points in 2 literature,4 points in 1 literature and 2 points in 1 literature.In all the RCTs,344 patients with dry eye were treated using sodium hyaluronate as the sodium hyaluronate eye drops group,and 342 dry eyes applied placebo,normal saline solution,hydroxymethyl cellulose,hydroxypropyl methyl cellulose respectively as the control group.Meta-analysis indicated that the total effective rate (OR) was 2.55 with the 95％ confidence interval (CI) 0.94-6.93.The symptoms were improved in 183 cases in the experiment group and 130 cases in the control group.No significant difference was found in the OR between the sodium hyaluronate eye drops group and the control group (Z =1.84,P=0.07).Conclusions Meta-analysis indicates that sodium hyaluronate eye drops can improve the symptom of dry eye,but have no significence with the control group.However,owing to fewer number and uneven quality of included RCTs,the large sample size and multicenter of RCTs is still needed to verify the superiority of sodium hyaluronate eye drops for dry eye.

Objective To investigate the expression of CDX2 in different subtypes of intestinal metaplasia (IM) and gastric cancer,and its relationship with Helicobacter Pylori (H.pylori) infection.Methods The expressions of CDX2 protein were detected with immunohistochemical method in 42 cases of chronic atrophic gastritis (CAG),46 cases of CAG with IM,34 cases of paracancerous IM and 50 cases ofgastric cancer.The IM was divided into three subtypes by HID-AB staining:27 cases of IM Ⅰ,23 cases of IM Ⅱ,and 30 cases of IM Ⅲ.H.pylori infection was detected with one minute rapid urease test,serum H,pylori IgG of ELISA method and HE staining in 80 caese of IM,which were divided into 46 cases of H.pylori-posi-tive groups and 34 cases of H.pylori-negative groups.Results The positive rates of H.pylori infection in IM Ⅰ,IM Ⅱ andIM Ⅲ were 66.67％,65.22％ and 43.34％,respectively,and there was no significant difference among different subtypes ofIM (x2=3.953,P＞0.05).The positive rates of CDX2 expression in IM Ⅰ,IM Ⅱ and IM Ⅲ were 85.19％,69.57％ and 36.67％,respectively,and IM Ⅲ were significant lower than IM Ⅰ,IM Ⅱ (x2 =13.899,P＜0.001;x2 =5.638,P=0.018),and there was no significant difference between IM Ⅰ and IM Ⅱ.Comparing of different types of IM group and gastric cancer group showed that the positive rates of CDX2 expression in IM Ⅰ,IM] were significant higher than in gastric cancer group (x2 =14.517,P＜0.001;x2 =5.509,P＜0.05),but there was no significant difference between IM Ⅲ and gastric cancer group (x2 =0.088,P＞0.05).The positive rates of CDX2 expression in H.pylori-positive groups was significant higher than in H.pylori-negative groups in all of.IM (76.09％ vs 44.12％,x2 8.525,P=0.004).Comparing between different subtypes of IM showed that the positive rates of CDX2 expression in H.pylori-positive groups was significant higher than in H.pylori-negative groups in IM Ⅲ (P=0.023),but there was no significant difference between IM Ⅰ and IM Ⅱ (P＞0.05).There was also no significant difference of CDX2 expression between H.pylori-positive groups and H.pylori-negative groups in gastric cancer.Conclusion H.pylori infection may affect the progression of IM and gastric carcinogenesis by affecting the expression of CDX2 in different subtypes of IM.

Objective To select valuable biomarkers for diagnosis and predicting outcome of sepsis-related acute respiratory distress syndrome(ARDS) from D-dimmer (DD),yon Willebrand factor (vWF),platelet (PLT),N terminal-pro brain natriuretic peptide (NT-ProBNP),interleukin-6 (IL-6),interleukin-8 (IL-8) and surfactant protein D (SP-D).Methods A total of 48 sepsis accompanied with ARDS patients and 40 sepsis patients were prospectively studied with comparison.The clinical characteristics of all the patients were recorded in detail.The blood samples were obtained within 24 hours of ICU admission.The concentration or activity of the seven biomarkers was quantitatively assayed and the results were recorded.To select the most valuable biomarkers as clinical indices,diagnosis model and death predictive model were constructed by Logistic regression.Results Among the seven candidate biomarkers,SP-D,vWF and IL-8 showed the most value.Their area under the receiver operator characteristic curve (ROC) were 0.758 (P ＜ 0.01),0.783 (P ＜ 0.01) and 0.747 (P ＜ 0.01) respectively,and raised to 0.847 (P ＜ 0.001) when the three biomarkers were combined.IL-8,age greater than or equal to 60 years and APACHE Ⅱ score greater than or equal to 20 were related to ARDS death with 12.138(lnIL-8)(P=0.022),6.157(P=0.040) and7.415(P=0.014) of OR values respectively.Conclusion SP-D,vWF,IL-8 should be valuable for early prediction of sepsis-induced ARDS and the diagnostic accuracy raised through combined utilization.IL-8 may be predictable for prognosis of sepsis related ARDS and the comprehensive evaluation combining clinical indices with IL-8 should be suggested in clinical practice.

The effect of reaction media cosolvent water activity, temperature and pH on Novozym 435-caulyzed enantioselective esterification of ketoprofen was systematically explored. Novozym 435 showed high catalytic activity and enantioselectivity in cyclohexane; E value increases markedly by addition of toluene to cyclohexane; the optimum temperature and the initial water activity were found to be 30℃ and 0.09 respectively; pH shows little effect on enzymatic reaction wilson the scope studied.

Objective To investigate drug resistance and genotypes of methicillin-resistant Staphylococcus aureus (MRSA) isolated from intensive care unit (ICU). Methods MRSA strains were isolated from patients, medical staff and environment of hospital ICUs. Disk diffusion (K-B method) was used for drug resistance testing; Staphylococcal cassette chromosome mec (SCCmec) and Staphylococcal protein A (spa) typing methods were used for genotyping and identifying the homology. Results There were 78 strains of Staphylococcus aureus isolated including 62 isolates of MRSA, which were mainly from the burn ICU (22, 35.48%). Among 62 MRSA strains, 50 were hospital acquired strains, in which 43 isolates were of SCCmec Ⅲ, 4 of SCCmec Ⅰ and 3 of SCCmec Ⅱ. Twelve isolates could not be typed. Twenty-eight out of 37 hospital acquired isolates were typed by spa typing as SCCmec Ⅲ-t030, which belonged to the same clone. Conclusion MRSA in ICU is multi-drug resistant and SCCmec Ⅲ-t030 is the most prevalent genotype, which indicates that clinical MRSA strains and environmental MRSA strains may be homologous.

ObjectiveTo investigate the effects of protein phosphatase 2A (PP2A) inhibitors on the viability of pancreatic cancer cell line PANC-1 and its mechanism.MethodsPANC-1 cells were treated with PP2A inhibitors Cantharidin or Okadiac acid.The activity degree of NF-κB pathway was tested by Western blot.NF-κB pathway was blocked from all sectors by PP2Acα plamid transfection,NF-κB inhibition of protein kinase α (IKKα) and NF-κB inhibitor α (IκBα) dominant negative mutant and p65 interfering plasmid.Cell viability was determined by MTT.ResultsPP2A inhibitors could induce phosphorylation of IKKα,further phosphorylation of IκBα and degradation and followed by the release of p65 into nucleus.When PP2Acα,IKKα dominant negative mutant and IκBα dominant negative mutant were overexpressed,or p65 was interfered,the inhibition rate of Cantharidin on cell viability decreased (31.85±13.37) ％,(23.48±8.98)％,(22.63±5.81)％ and (20.88±3.24)％respectively,and the inhibition rate of Okadiac acid on cell viability decreased (40.17 ± 11.65)％,(27.34±14.28)％,(24.85±3.39)％ and (27.08±3.81)％ respectively.ConclusionsPP2Ainhibitors play a role in preventing pancreatic cancer through PP2Acα/IKKα/IκBα/p65 pathway.