This study analyzed the genetic evolution of neuraminidases (NAs) of influenza A subtype N9 viruses with the aim of determining the genetic origin of the novel avian A/H7N9 influenza virus. The NA sequences of influenza A subtype N9 viruses available from NCBI were used to construct a phylogenetic tree using the programs ClustalX 2.0 and MEGA 6.0. This analysis indicated that the novel avian A/H7N9 influenza virus is located in the modern Eurasian phylogenetic cluster. This cluster was then further analyzed by estimating the overall rate of evolutionary change and the selective pressure at the nucleotide level using the program BEAST 2.1.2 and the web interface Datamonkey, and by generating an amino acid sequence entropy plot using Bioedit software. In this cluster, the mean rate of nucleotide substitutions in NA was found to be 3.8354 x 10(-3) and the mean ratio of non-synonymous (dN) to synonymous (dS) substitutions per site (dN/dS) was 0.140413. A particularly high level of amino acid mutation entropy was identified at nucleotides 16, 19, 40, 53, 81, 84, 112, 256, 335, 359, and 401. This genetic evolution analysis suggests that the nucleotide substitutions that characterize the novel avian A/H7N9 influenza virus neuraminidase are likely to result from the overall genetic evolution of influenza A subtype N9 virus NAs, and not from selective stress. Phylogenetic analysis suggests that the influenza A virus (A/duck/Siberia/700/1996(H11N9)) isolated in 1996 appears to be the common ancestor of the more recent influenza A subtype N9 viruses NAs.
Amino Acid Substitution ; Animals ; Birds ; Evolution, Molecular ; Humans ; Influenza A Virus, H7N9 Subtype ; chemistry ; enzymology ; genetics ; Influenza in Birds ; virology ; Influenza, Human ; virology ; Molecular Sequence Data ; Neuraminidase ; chemistry ; genetics ; Phylogeny ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

Objective:To generate rabbit polyclonal antibody against human Argonaute2 (Ago2) protein and to identify its functional characterization for determination of differential expression and cellular localization of Ago2 protein in various cell lines.Methods:DNAstar software was applied for searching the high antigenicity region of Ago2 gene sequence termed k-Ago2.Prokaryotic expressing plasmid was constructed and transformed to E.coli BL21 (DE3) to induce expression by IPTG.The fusion protein was injected into rabbits subcutaneously to produce polyclonal antibodies after purification by gel regaining.ELISA was operated to detect antibody titer.Western blot was used to identify the specificity and sensitivity of the antibodies and detect the differential expression of Ago2 protein in various cell lines.Meanwhile,immunofluorescence experiments were arranged to show cellular localization of Ago2 protein.Results:The prokaryotic expressing plasmid was constructed correctly.K-Ago2 protein was expressed and purified,and then rabbit polyclonal antibodies against Ago2 were generated after immunization with k-Ago2 protein.The titer detected by ELISA was 1∶19 000.Western blot results demonstrated the high specificity of the antibodies.Finally,we successfully observed the differential expression and cellular localization of Ago2 protein in various cell lines.Conclusion:The polyclonal antibody against Ago2 protein has been achieved successfully.It will be propitious for the intensive study of the RNAi mechanism and even profound clinical application.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Child, Preschool ; Combined Modality Therapy ; Cord Blood Stem Cell Transplantation ; adverse effects ; Female ; Graft vs Host Disease ; therapy ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; therapy ; Transplantation, Homologous

Adolescent ; Child ; Child, Preschool ; Clinical Trials as Topic ; Flow Cytometry ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; diagnosis ; pathology ; therapy ; Neoplasm, Residual ; diagnosis ; pathology ; therapy ; Polymerase Chain Reaction ; Prognosis ; Recurrence ; Sensitivity and Specificity

Adolescent ; Female ; Humans ; Lung Neoplasms ; congenital ; Lymphoma, Large-Cell, Anaplastic ; physiopathology

Aim: To synthesize the ent-kaurene diterpenoid and its derivatives from natural available stevioside.Methods: The crucial pharmacophore of exo-methylene ketone was designed and the carboxyl group at C-4 was modified.The cytotoxic inhibition activities in vitro of target compounds were evaluated against human cancer cells BEL-7402,HO-8910,MCF-7 and HL-60 by a MTT method.Results and Conclusion: The struc-tures of new target compounds were identified by ~1H NMR,FT-IR and EI-MS.The preliminary experimental re-sults showed that some derivatives of ent-kaurene diterpenoid possessed fair inhibitory activity against tumor cell lines compared with that of their parent steviol.

Diabetic Retinopathy ; complications ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Retinal Vein Occlusion ; complications ; Vitrectomy ; Vitreous Hemorrhage ; etiology ; surgery

AIM: To evaluate the effects of total flavonoid of chrysanthemum on the expressions of Fas and FasL in male rabbits with dry eye, and to investigate the therapeutic effects of the total flavonoid of chrysanthemum on dry eye.
METHODS: Totally 150 male Japanses white rabbits were divided into blank group ( group A ) , sham -operated group ( group B ) , model group ( group C ) , androgen control treatment group (group D), and total flavonoid of chrysanthemum treatment group ( group E ) . The dry eye model was established with orchiectomy on group C, D and E. Rabbits in group E were treated with total flavonoid of chrysanthemum. Rabbits in group D were treated with androgen intramuscular injection. Rabbits in the group A, group B, group C was treated with normal saline. All rabbits were detected with Schirmer's Ⅰ test and tear break-up time (BUT). Fas, FasL were checked on immunohistochemistry.
RESULTS:The Schirmer's I test values of group E was significantly higher than that of group C ( P<0. 01 ) and the BUT value of group E was significantly longer than that of group C ( P<0. 01 ). The quantity of positive expression of Fas in glandular tube cell and acinar epithelial celland apoptosis cells of group E after treatment at 1, 3, 5mo were significantly lower than that of group C, cell population of the positive expression of FasL was obviously higher than that of group C (P<0. 01).
CONCLUSION:The main component of chrysanthemum is flavonoid, which could significantly inhibit happening of dry eye in rabbit after androgen level lowered and lacrimal gland apoptosis and keep basic tears secretory volume and tear film stability.

Objective To understand the reliability and validity on SF-36 scale and the quality of life (QoL) among patients with advanced-schistosomiasis in Jiangsu province.Methods Cluster sampling method was used to choose 586 advanced-schistosomiasis patients who were registered in Jiangsu province.Questionnaire including SF-36 module,was used for face to face interview on the patients.Statistical analyses were made to assess the results,including correlation analysis,reliability analysis,factor analysis,t-test and one-way ANOVA.Results The split-half reliability coefficient was 0.92,P＜0.001 and the internal consistency reliability was 0.90,with the range of Cronbach' α coefficient on each dimension was from 0.69 to 0.98.Through principal component analysis,two factors were identified,with the accumulative contribution rate as 67.37％.SF-36 showed certain discriminant validity to distinguish groups in different economic levels.Compared with the domestic norm data,the scores of SF-36 on those patients were generally lows.Conclusion SF-36 had good reliability and good criterion validity,with a certain degree of discriminant validity,but with poor constructs validity.Level of QoL of patients with advanced schistosomiasis in Jiangsu was low because of being chronically ill,that called for more strategies to improve the QoL of patients.

Objective To investigate the experience of endoscopic mini-invasive therapy for residual lesions of peripancreatic necrotizing infection with choledocoscopy-assisted debridement technique, and to explore its clinical application value. Methods 71 patients with postoperative surgical drainage and accompanied with residual focus were collected. Choledochoscope was inserted via the drainage sinus, and the focus was observed and necrotic tissue was removed under direct choledochoscopic vision. Results Of the 71 patients who underwent this procedure, 64 were cured (success rate, 90.1%); 3 patients withdraw from treatment due to economic reasons; 4 patients received open surgery after 1 ～ 3 times of choledocoscopy-assisted debridement. The 64 cured patients received 2 ～ 9 times of choledocoscopy-assisted debridement with a mean of 5.1 times. 87.5% patients needed 4 ～ 6 times of procedures. The healing time was 18 ～ 125 days (average 71.3 days). Hemorrhage occurred in 3 patients and digestive tract fistula occurred in 2 patients and were resolved with non-operative management. Conclusions With the help of postoperative established surgical drainage channel, choledochoscopy-assisted debridement could be considered as a safe and effective miniinvasive treatment for residual focus of peripancreatic necrotizing infection, and is worth of clinical application.