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 investigate the effects of miR-135a on HOXA10 expression,proliferation and apoptosis of SKOV3 cells.Methods (1) Through computer-aided algorithms,the predicted target gene of miR-135a (HOXA10)were determined.(2) miR-135a mimics,miR-135a inhibitor and negative control were transfected into SKOV3 cells,respectively.Reverse transcription (RT)-PCR,western blot analysis were used to examine the expression levels of HOXA10 at different times (24,48 and 72 hours).(3) A luciferase reporter assay was used to confirm the direct regulation between miR-135a and HOXA10.(4) SKOV3 cells proliferation at different times (24,48 and 72 hours) was detected by methyl thiazolyl tetrazolium(MTT) assay [quantified by absorbance(A)].Western blot was used to examine the expression of apoptosis-associated protein bcl-2,bax and caspase-3 in SKOV3 cells after 48 hours transfection.Results (1) HOXA10 was predicted to be the target gene of miR-135a by computer-aided algorithms.(2) RT-PCR shown that HOXA10 mRNA levels were decreased over time (24,48 and 72 hours) after miR-135a mimics transfectionin SKOV3 cells (0.94 ±0.04 vs 0.78 ±0.03 vs 0.70 ±0.03,P ＜0.05).While,the expression of HOXA10 mRNA was increased over time after miR-135a inhibitor transfection (1.14 ± 0.05 vs 1.16 ±0.03 vs 2.60 ±0.08,P ＜0.05).After transfected with miR-135a mimics or miR-135a inhibitor over 48 and 72 hours,the HOXA10 expression levels in SKOV3 cells were significantly lower or higher than each control group,respectively (all P ＜ 0.01).Western blot analysis of HOXA10 expression in SKOV3 cells confirmed the results of RT-PCR detected.(3) After cotransfection of miR-135a plasmid and pMIR-REPORT luciferase plasmid containing HOXA10,luciferase reporter assays showed that the luciferase activity reduced by 67.8％ (P ＜0.01).(4) MTT showed that SKOV3 cells growth after miR-135a mimics transfection for 48 and 72 hours were significantly lower than those in control group (0.38 ± 0.03 vs 0.52 ± 0.05,0.67 ±0.05 vs 0.75 ± 0.06 ; respectively,all P ＜ 0.05).While,SKOV3 cells transfected with miR-135a inhibitor for 72 hours grew significantly faster than that in control group (0.95 ± 0.05 vs 0.75 ± 0.06,P ＜ 0.01).After miR-135a mimics transfection,the level of bcl-2 protein was significantly lower than that in control group (0.28 ±0.06 vs 0.76 ±0.09,P ＜0.01).The activity of caspase-3 was significantly higher than that in control group (115.0 ± 2.4 vs 95.4 ± 2.1,P ＜ 0.01).While,there was no statistical difference of bax expression (P =0.142).However,after miR-135a inhibitor transfection,the expression level of bcl-2 protein was significantly higher than that in control group (0.92 ± 0.03 vs 0.76 ± 0.09,P =0.037) and the activity of caspase-3 was significantly lower than that in control group (59.5 ± 4.1 vs 95.4 ± 2.1,P ＜ 0.01).There was also no statistical difference of bax expression (P =0.066).Conclusion miR-135a may play an important role in cell proliferation and apoptosis of ovarian cancer cells by regulating HOXA10 and its downstream pathways.

Objective To analyze the clinical characteristics of dopa-responsive dystonia (DRD). Methods The clinical characteristics and the causes of misdiagnosis in 15 patients with DRD were reviewed retrospectively.Results 4 male and 11 female patients were investigated in this study. The age at DRD onset was from 3~25 years old,the average age was 11.7 years old.The courses ranged from 2 to 29 years and the average was 12.1 years. The symptoms in female patients often aggravated during gravidity and delivery. Main clinical features included gait irregularity, postural instability and tremor, with marked diurnal fluctuation. The disease may be misdiagnosed as cerebral palsy, idiopathic torsion dystonia, essential tremor, parkinson's disease, neurosis and so on. All the patients in this study improved obviously in 2 weeks after therapy with low dose of Madopar, and recovered in 2~4 years of follow up.Conclusions Clinical diversity and diurnal fluctuation are the characteristics of DRD. Early trial treatment with low dose of Levodopa should be highly recommended for the generalized dystonia in children and untypical adults.

SYBR Green I Real Time-qPCR method was developed to quantify the numbers of copyies of AlkB ( alkanes degradation gene) and Nah ( naphthalene dioxygenase degradation gene) functional degradation gene corresponding to alkanes and aromatic hydrocarbons degradation. Two pairs of primers AlkBf/AlkBr and Nahf/Nahr were designed for AlkB and Nah amplification respectively, according to the nucleotide sequences of related degradation microorganisms published in GenBank. The purified recovery products of traditional PCR were combined with pEASY-T1 vectors and transformed in competent cells to amplify. The recombinant plasmids were extracted and used as positive templates to create standard curve through gradient dilution. The conditions for the real time PCR were as the follows: the final concentration of forward and reverse primers were 0. 2 μmol/L, 2×TransStart Top Green qPCR SuperMix, and the annealing temperatures of AlkB and Nah PCR were 50℃ and 57℃, respectively. The method showed a sensitivity of 100 times higher than that of the traditional PCR method and good repeatability. The numbers of copies of AlkB in three functional regions of an oilfield indicated that oil producing zone with serious oil pollution had the highest AlkB copy numbers, and residential zone with lighter oil pollution had the lowest AlkB copy numbers. Nah degradation gene distribution was more uniform.

asty without increasing morbidity, especially for slim patients or patients with a large renal pelvis.

Retinal neovascularization is a complicated pathophysiological process as a result of imbalance between angiogenic and anti-angiogenic factors. Correct understanding of the signaling pathways,exploring the critical factors involved in retinal angiogenesis, looking for new strategies by reconstructing the new vessels are helpful for knowing the mechanism of the occurrence and development of reitnal neovascularization, which would be good for preventing and treating retinal neovascularization diseases.

Nowadays, various voice disorders are sprung out towards teachers of primary and secondary school, affecting their personal life and social work. Through literature review, the author aims at clearing up series of assessments and methods of voice training which are suitable for the teachers of primary and secondary school, such as establishing a right way of respiratory, phonation and resonance, avoiding occupational risk factors such as vocal abuse or misuse to regain healthy voice. Review of the several relevant literature and commentary about voice disorders of the teachers in primary and secondary school. Although various methods of voice training were reported from time to time to guide us SLP to carry out voice therapy, however, deviations of the clinical data from scholars' different points of view are waiting for our further support through case-control study.
Faculty ; Humans ; Schools ; Voice Disorders ; diagnosis ; rehabilitation

Objective: To revise and evaluate the Chinese version of the FACT-B. Method: The Chinese version of the FACT-B was translated from the original one, after back-translated and culture adaptation, it was applied to 165 patients with breast cancer. Result: The test-retest reliability of five domains, such as physical well-being, social/family well-being, emotional well-being, functional well-being, additional concerns and the overall scale were 0.82～0.89. The internal consistency alpha for the five domains were 0.61～0.84. After admission to hospital 2-4 weeks, the quality of life of patients showed some change in this scale. Conclusion: The Chinese version of the FACT-B has good psychometric quality and can be used to assess the quality of life of Chinese patients with breast cancer.

The pharmacological characteristics of recipes are multi-components, multi-ways, multi-links, and multi-targets, on which the efficacy substances in recipes are worth of studying. To explore the essential substances of Chinese herbs in recipe is one of the key points for modernization of Chinese materia medica. The pharmacological effects and bioactivities inside the target tissue of lesion body are still not clear, even though there are some progress in herb serum pharmacology, recipe pharmacokinetics, and pharmacodynamics. The main effective components of recipe to pancreas in acute pancreatitis models by ig or enema were investigated by HPLC-MS method so as to establish a new hypothesis (tissue pharmacology of recipe): i.e. guided hy fundamental theory of traditional Chinese medicine to study the effect and the mechanism of recipe's essential substances on target tissues with pharmacological methods; to establish the experimental method for searching the foundation, the principle, and the direct substances of recipes which act on target tissues; to form more effective formula if possible, and to explore in a new researching field for the recipes of Chinese materia medica.

Ojective To investigate the relationship between the serum adiponectin concentration and insulin resistance in patients with obesity and diabetic patients. Methods Fasting serum adiponectin concentration was measured by RIA in 19 normal subjects, 23 obese subjects, 31 diabetic patients without obesity and 26 diabetic patients with obesity. Results There was no significant difference in serum adiponectin concentration between nomal and obese subjects, and between type 2 diabetics without obesity and type 2 diabetics with obesity . The serum adiponectin concentrations was significantly higher in type 2 diabetics without or with obesity than that in nomal and obese subjects. There was significant difference in serum adiponectin concentration between female and male subjects. Body mass index (BMI), waist circumference(WC), waist-hip ratio (WHR), body fat percentage (BF%), fasting plasma glucose (FPG), insulin sensitivity index (ISI), triglyceride (TG) and high density lipoprotein-C (HDL-C) were significantly correlated with serum adiponectin concentration. A stepwise mulitiple linear regression analysis showed that HDL-C, FPG, WC, BF% entered the regression equation. Conclusion Serum adiponection level was associated with sex, obesity, type 2 diabetes mellitus and insulin resistance.