A simple analytical method by means of on-line solid phase extraction followed liquid chromatography-tandem mass spectrometry ( SPE-LC-MS/MS) was developed for the simultaneous quantitation of 4 endocrine disruptors ( triclosan, triclocarban, bisphenol A and nonylphenol) in dairy products. Infant formula and milk samples were dissolved in acetic acid buffer and hydrolyzed by β-glucuronidase/arylsulfatase. Acetonitrile was used as the extract. Then, the mixture was freeze-centrifuged for 10 min and the supernatant was diluted with water, and analyzed via on-line SPE-LC-MS/MS. The sample extracts were concentrated by an Xbridge C8 cartridge and separated on a BEH C18 column with a gradient mobile phase of methanol and water; then analyzed by triple quadrupole mass spectrometry. Mass acquisition was conducted under negative electrospray ionization mode. Quantification was performed by isotopic internal standard calibration. Acceptable linearity (R2>0. 99) was achieved over the range of 0. 005-5. 0 μg/L, with limits of quantification of 0. 03-1. 0μg/kg. Average recoveries of four target compounds (spiked at three concentration levels) ranged from 80. 2%-106. 7%,with relative standard deviation less than 15%. Due to its rapidity, simplicity, and high sensitivity, the method is suitable for the analysis of endocrine disruptors in dairy products. It has been applied in the analysis of raw milk and milk products collected in Beijing. As a result, nonylphenol was found with a high detectable frequency.

Objective　To obtain genetic characterization of the HA1 of new isolates of influenza A （H1N1） virus. Methods　Virus was amplified in embryonated chicken eggs， the virion DNA was transcribed into cDNA by reverse transcriptase， cDNA amplified by PCR， the products of PCR were purified. Afterward， DNA sequence analysis was performed by the dideoxy-mediated chain termination method， using synthetic oligo nucleotide primers. Results　The HA1 domain of new isolates of influenza A （H1N1） virus showed that their HA1 genes were 981 nucleotide in length coding for a HA1 protein with 327 amino acids， deletion of a glycosylation site and an amino acid. The homology of amino acid sequences of protein molecules on HA1 domains of new influenza A virus when compared with A／Guifang／10／94（H1N1） and A／Bayern ／07／95 （H1N1） viruses，was 92.8％ and 91.3％ respectively； The homology of amino acid sequences of protein molecules on HA1 domains of A／Jingjun／11／98（H1N1） when compared with A／Jingjun／13／98（H1N1） or A／Jingjun／807／97（H1N1） viruses，was as high as 98％； The homology of amino acid sequences of protein molecules on HA1 domains of A／Guifang／10／94（H1N1） when compared with A／Bayern／07／95（H1N1） viruses was as high as 96％. Conclusions　The HA1 gene of　new H1N1 virus strains is　different from those of A／Guifang／10／94 （H1N1） and A／Bayern／07／95（H1N1）， they will probably be new mutation strains.

Body fat of 12 male adults were measured by water displacement me-thod(density method) at every morning for 5 successive days. The standard deviation of single observation was 0.29kg calculated by mean residual lung volume method. It was significantly lower than the value (0.5kg) calculated by the ordinary method (p

Objective:To evaluate the importance of 3D-CTA with volume rendering for the diagnosis of multiple intracranial aneurysms. Methods: Axial source images were obtained by helical CT scanning and reconstruction of 3D-CTA images was done by volume rendering technique in conjunction with multiplanar reformation. Results: In the past one year,there were 10 patients diagnosed as having multiple intracranial aneurysms by 3D-CTA and altogether 24 aneurysms were visualized,including 10 small aneurysms(≤5mm.Three dimensional CT angiography with volume rendering demonstrated aneurysms very well and provided useful information concerning the site,shape,size and spatial relationship with the surrounding vessels and bone anatomy. Conclusion: Three-dimensional CT angiography with volume rendering is a quick,reliable,and relatively noninvasive method for diagnosing multiple intracranial aneurysms.It delineates detailed aneurysmal morphology,and provides useful information for planning microsurgical approaches.

Objective To determine which parameters of reverse remodeling of left ventricular could become the effective indexes in evaluating the short-term therapeutic effect of cardiac resynchronization therapy(CRT)in heart failure(HF)patients.Methods CRT was performed in 26 HF patients with dysfunctions of wall motion.Serial echocardiography was practiced at baseline,one and three months after CRT.The parameters including left atrial end-diastolic diameter(LADD),left ventrieular end-diastolic diameter(LVDD),end-diastolic volume (LVEDV),end-systolic volume(LVESV),ejection fraction(LVEF)were measured and compared before and after CRT therapy.Results At one and three months after CRT,respectively,CRT was associated with reduced LADD,LVDD,LVEDV,LVESV and improved LVEF and filling time compared with those at baseline.Moreover,all these parameters had better correlations with the activities of these patients.Conclusions LADD,LVDD,LVEDV,LVESV and LVEF could become the effective indexes in evaluating the short-term effect of CRT in HF patients.The reduced diameters and volumes of left atria and left ventricle are more sensitive parameters than the improved LVEF.

Objective Myelodysplastic spndrome (MDS) patients' chromosome aberrations were detected by CC and FISH method respectively,to establish standard FISH platform of diagnosis of MDS,to compare the differences of FISH and CC in the detection of MDS chromosome aberrations.Methods Chinese MDS patients fulfilled with the Vienna minimum diagnostic criteria were tested by CC and FISH method,and the patients' clinical data was collected at the same time.Results The standard FISH platform of diagnosis of MDS was established successfully,including probe combination of-5/5q-,+8,-7/7q-,20q-,-Y,the success rate was 100 ％.In 83 cases of MDS,chromosome aberrations rate detected by two methods was 42.2 ％,30.1％ of CC method and 33.7 ％ of FISH method,the accordant diagnostic rate was 76.1％.FISH had higher positive rate than CC method (29.6 ％ vs 18.6 ％,P =0.049) in the IPSS intermedian-risk 1 group,but lower positive rate in IPSS intermedian-risk 2 group (62.5 ％ vs 81.2 ％,P =0.036).There were 11 (19.0 ％) cases of clonal chromosome aberrations detected by FISH method but not detected by CC method,in which 10 (90.9 ％) cases were intermedian-risk 1 group (IPSS grouping).Evaluate IPSS by FISH and CC respectively,the accordant rate was 86.7 ％.Evaluate IPSS by CC alone will result in 3 patients (3.6 ％)decline in risk stratification,and evaluate IPSS by FISH alone will result in 8 patients (9.6 ％) decline in risk stratification.There were 35 (42.2 ％) cases of chromosome aberrations detected by the two methods,including 13 (15.7 ％) cases of complex karyotype and 22 (26.5 ％) cases of single abnormal karyotype.The most common chromosomal aberration was +8,a total of 9 (10.8 ％) cases,followed by abnormalities of chromosome 7,a total of 7 (8.4 ％) cases.Conclusion The study has established standard FISH platform of diagnosis of MDS,the method is stable,sensitive and rapid.FISH technology joint CC technology can improve the patients with MDS chromosome aberration detection rate.Compared to CC,FISH technique can improve the detection rate of the intermedian-risk 1 IPSS grouping MDS patients.Because of limited probe,FISH technique can not entirely replace the role of CC method in MDS,but can complement each other.

Objective To investigate the clinical characteristics and prognostic factors in patients with primary gastric non-Hodgkin lymphoma (PG-NHL).Methods The pathological data of 51 PG-NHL patients admitted in our hospital from 2003 to 2013 were analyzed retrospectively.Results In 51 patients with PG-NHL,there were 26 males and 25 females.The patients' age ranged from 18 to 80 years old with median age as 56 years old.The median survival time was 32 months (range from 1 to 114 months).The oneyear overall survival (OS),three-year OS and five-year OS were 90.2 ％,82.4 ％ and 80.4 ％,respectively.The surgery did not significantly improve PG-NHL patients' progress free survival and OS.Only 1 (2.0 ％) patient had gastrointestinal hemorrhage and perforation after chemotherapy.However,6 (46.2 ％) patients suffered from early satiety,gastric emptying disorder,alkaline reflux gastritis and dumping syndrome in surgery group.Conclusions Surgery did not improve the survival of PG-NHL patients.The life quality in chemotherapygroup is better than that in surgical group.

Objective:To study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs.Methods:BABL/c mice were randomly divided into VLPs experimental group, alum adjuvant experimental group, PBS control group and alum adjuvant control group,the experimental group mice were intramuscular immunization with HBoV1 VP2 VLPs and HBoV1 VP2 VLPs added alum,control group mice were immunization with alum or PBS buffer,then to study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs by cellular and humoral immune strength.Results: Alum adjuvant decreased cellular immune response induced by HBoV1 VP2 VLPs(P<0.001),enhance the HBoV1 VP2 VLPs immuned serum IgG titer(P<0.05)and IgG activity(P<0.01).Conclusion:Alum adjuvant can enhance humoral immune response induced by HBoV1 VP2 VLPs,but weaken cellular immune response induced by HBoV1 VP2 VLPs,when HBoV1 VP2 VLPs used as a prophylactic vaccine it should add alum adjuvant,when used as a therapeutic vaccine,it should not add alum adjuvant.

To investigate the effects of component I from Agkistrodon acutus venom (AAVC-I) on the biological features of chronic myeloid leukemia cells, K562/A02 leukemia cells were cultured in the presence of AAVC-I (6.25 - 100 µg/ml) and the proliferation status was assayed by CCK-8 method. Morphological changes were observed by inversed microscope after Giemsa and Hochest 33258 staining, and cell apoptosis was detected by flow cytometry. Caspase 3 activity was tested by using Chromogenic Activity Assay Kit. The results showed that AAVC-I inhibited the growth of K562/A02 cells in time- and concentration-dependant manners, and the IC(50) at 48 h was 30.988 µg/ml. Giemsa and Hochest 33258 staining showed the typical apoptotic features in K562/A02 cells after induction with AAVC-I for 48 h. Flow cytometric analysis revealed that the percentage of the apoptotic cells reached from 0.88 up to 53.66 as the treated concentration was elevated from 0 to 50 µg/ml. Compared with the control group, the expression of caspase 3 in the tested group was enhanced in a dose-dependent manner (P < 0.05). It is concluded that AAVC-I can effectively inhibit the growth and promote apoptosis of K562/A02 cells. Elevated expression of caspase-3 may be attributed to the apoptosis of K562/A02 cells.
Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Crotalid Venoms ; pharmacology ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Leukemia ; metabolism ; pathology

ObjectiveNotch signaling is highly conservative in evolution and plays an important role in cell's proliferation and differentiation.Construction of tentiviral vector containing Notch intracellular domain (NICD) would lay the foundation for the study of Notch signaling.MethodsTotal RNA was extracted from the myeloid tissue of C57BL/6 mice.cDNA was composed via reverse transcription.NICD sequence was obtained by PCR and recombined into lentiviral vector.Lentiviral vector with NICD was infected into target HEK293T cell.Real-time PCR and Western blot were used to examine NICD expression in HEK293T cell.ResultsNICD expression increased in HEK293T cells.ConclusionThe successful construction of lentiviral vector involving mice NICD expression provides the foundation for the future study.