This study is to establish a cell-based model targeting to neuraminidase (NA) of the 2009 H1N1 influenza A virus. NA is an influenza virus structural protein with enzymatic activity of the cleavage of HA-sialic acid interaction to release new viral particles from cells. A model of HIV-1 (pNL4-3.Luc.R(-)E(-)) based pseudovirions packed with HA [hemagglutinin, A/VietNam/1203/2004 (H5N1)] and NA [A/California/04/2009 (H1N1)] was established to evaluate compounds activities on NA function. The viral release can be blocked by neuraminidase inhibitors, oseltamivir and oseltamivir carboxylate, with IC50 of (61 +/- 31) nmol L(-1) and (5.5 +/- 2.9) nmol L(-1) respectively. A point mutation of H275Y on NA leads oseltamivir-resistance. This corresponding mutation was introduced into the system which was also confirmed by oseltamivir and oseltamivir carboxylate.

Cytomegalovirus Infections ; complications ; Female ; Humans ; Infant, Newborn ; Male ; Skin Diseases ; etiology

Objective To evaluate efficacy and safety of glucocorticoids for community-acquired pneumonia (CAP).Methods We searched relevant randomized controlled trials (RCT) from Pubmed,EMBASE,Cochrane Library,Chinese Journal Full-text Database,Chinese Biomedical Literature Database and traced the related reference to obtain the information that had not been found by using key words of steroids,glucocorticoids,cortisol,corticosteroids,community acquired pneumonia and CAP.The quality of RCT included was evaluated.Meta-analysis was performed with RevMan 5.0 provided by the Cochrance Collaboration.Results Seven RCTs involving 944 patients were included in the meta-analysis.Mean length of hospital stay in glucocorticoids treatment group was significant shorter than that in conventional treatment group (WMD =-1.70,95％ CI:-2.01-1.39,Z =10.81,P ＜0.01).No significant differences were found regarding mortality (RR =0.77,95％ CI 0.46-1.27,Z =1.03,P =0.30),mean length of ICU stay (WMD =1.17,95％ CI:-1.68-4.02,Z =0.81,P =0.42),the risk of double infection (RR =1.32,95％ CI 0.66-2.63,Z=0.79,P=0.43),the incidence of hyperglycemia (RR=1.84,95％ CI0.76-4.41,,Z=1.36,P=0.17),the incidence of upper gastrointestinal bleeding (RR =1.98,95％ CI O.37-10.59,Z =0.80,P =0.42) between conventional treatment group and glucocorticoids treatment group.Conclusions Glucocorticoids treatment used in patients with community-acquired pneumonia may shorten the course of illness and possess better safety,but could not reduce the mortality.

Objective To know basic characteristics of MSM infected with HIV treated by highly active antiretroviral thera‐py(HAART) ,and evaluate therapeutic effect based on the level of CD4+ T cell counts .Methods Basic database of patients were collected from the national AIDS comprehensive data information system ,patients were observed by the retrospective cohort study , and effect indicators were analyzed based on the level of CD4+ T cell counts .The data was analyzed by SPSS18 .0 software .Results A total of 1 758 cases were recruited in this study and the average age was (33 .5 ± 11 .1)years .The component ratio of the CD4+T cell counts <50/μL ,50 to 199/μL ,200 to 350/μL ,> 350/μL were 7 .7% ,27 .6% ,54 .9% ,9 .7% ,respectively .The CD4+ T counts increased and kept among 150 to 250/μL ,250 to 350/μL ,350 to 450/μL ,450 to 550/μL ,respectively after 12 months ,treat‐ment .The rates of viral inhibition were 70 .2% ,81 .7% ,84 .2% ,83 .3% ,respectively after 6 months′treatment .And the cumulative survival rates were 0 .81 ,0 .93 ,0 .98 ,1 .0 after 36 ,months treatment ,respectively .Conclusion HAART on MSM achieves remarka‐ble results .And the early treatment was more beneficial to improve immune function ,reduce AIDS mortality and control the HIV/AIDS epidemic of MSM .

Objective To assess the biophysical characteristics of normal skin and inflammatory skin lesions in mild and moderate acne. Methods Seventy-five mild and moderate acne vulgaris patients were included in the study. One inflammatory lesion measuring 2.0 to 5.0 mm in diameter which occurred within 48 hours prior to the measurement was selected as the target lesion. Trans-epidermis water loss (TEWL), capacitance and a* value were measured in target lesions and lesion-adjacent normal skin. Sebum content immediately,1 and 4 (saturated) hours after face washing was also determined on target lesions and central forehead between the eyebrows. Sebum secretion rate (SSR) was calculated. Results The TEWL and SSR significantly increased in the lesion-adjacent normal skin with the increment of inflammatory lesion number (both P < 0.05), and increased in target lesions with the elevation of a* value (both P < 0.05). The target lesions exhibited a significantly higher TEWL but a lower saturated sebum content and SSR than the adjacent normal skin did (all P <0.05). Conclusions The severity of inflammatory lesions is correlated with the impaired barrier function and increased SSR in facial skin. Compared with the adjacent normal skin, inflammatory skin lesions have a reduced skin barrier function and SSR.

Ginsenosides,which belongs to triterpenoid saponins of plant terpenoids,are the main active components of the valuable medicinal herbs ginseng and American ginseng.Recent studies show that ginsenosides have a variety of beneficial effects,including anti-inflamatory,antioxidant,and anticancer effects.Ginsenosides are synthesized by complicated modification of triterpenoid skeleton after cyclization of 2,3-oxidosqualene through triterpene saponin synthesis pathway in which they share common early steps with phytosterols synthesis.It outlines recent advances for the biosynthesis of ginsenosides,the cloning and characterization of genes encoding key enzymes in the pathway and the basal framework of ginsenosides biosynthesis pathway.The prospects of secondary metabolism engineering in the biosynthesis of plant natural products and its application in ginsenosides biosynthesis are also discussed.

Southern blot analysis with probe from mini-Tn5 gfp-km transposon indicated that 5 non-pathogenic mutants which were generated by insertion of mini-Tn5 gfp-km mutagenesis contained a single copy of the transposon. Using genomic DNA of each mutant as a template, TAIL-PCR was performed with seven arbitrary degenerate (AD) primers pairing with 3 nested specific primers designed based on the sequence of GFP toward outside in mini-Tn5 gfp-km. After 3-step PCR reactions, the flanking sequence of each mutant was obtained. The PCR product was ligated with pGEM-T EASY vector and then was transformed into E. coli DH5 alpha by electroporation. Positive clones were selected by white/blue colony and plasmid was isolated, then digested with EcoRI. Plasmid was sequenced if its insert was longer than 300 bp. Our results indicated that TAIL-PCR was proved to be a simple and efficient approach in identification of gene using insertion mutagenesis.
Base Sequence ; Cloning, Molecular ; methods ; DNA Transposable Elements ; DNA, Bacterial ; isolation & purification ; Genes, Bacterial ; Green Fluorescent Proteins ; Luminescent Proteins ; genetics ; Molecular Sequence Data ; Mutagenesis ; Polymerase Chain Reaction ; methods ; Xanthomonas campestris ; genetics ; pathogenicity

Mean hemoglobin (Hb) concentration of about 3 500 subjects derived from 17 studies of Himalayan highlanders (Tibetans, Sherpas, and Ladakhis) was compared with lowlanders (Chinese Han, Indian Tamils) lived in the Himalayas, and European climbers during Everest expeditions as well as Andean natives. The results found that Hb concentration in Himalayan highlanders was systemically lower than those reported for Andean natives and lowland immigrants. These comparative data demonstrated that a healthy native population may successfully reside at high altitude without a significant elevation in Hb, and the lower Hb levels of Himalayan highlanders than those of migrated lowlanders and Andean natives are an example of favourable adaptation over the generations. In addition, excessive polycythemia has frequently been used as a marker of chronic mountain sickness (CMS). Altitude populations who have a higher Hb concentration also have a higher incidence of CMS. The low Hb in Himalayans suggested as showing adaptation over many generations in Tibetan stock. Recent work in Tibet, suggested that Tibetans there may have adapted to high altitude as a result of evolutionary pressure selecting for genes which give an advantage at altitude. All of the population genomic and statistical analysis indicated that EPAS1 and EGLN1 are mostly likely responsible for high altitude adaptation and closely related to low Hb concentration in Tibetans. These data supported the hypothesis that Himalayan highlanders have evolved a genetically different erythropoietic response to chronic hypoxia by virtue of their much longer exposure to high altitude.
Adaptation, Physiological ; Altitude ; Asian Continental Ancestry Group ; genetics ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Evolution, Molecular ; Hemoglobins ; genetics ; Humans ; Hypoxia-Inducible Factor-Proline Dioxygenases ; genetics ; Tibet

Objective To investigate whether high glucose can induce endothelial-mesenchymal transition (EndMT) in glomerular endothelial cells and the role of TGF-β in the process.Methods Rat glomerular endothelial cells were divided into five groups:normal glucose (NG,5.5mmol/L),high glucose (HG,15,30 mmol/L),TGF-β inhibition (HG+ LY36,30 mmol/L glucose + 10 μmol/L LY364947),hyperosmotic control (M,5.5 mmol/L glucose+25.5 mmol/L mannitol) and solvent control (D,5.5 mmol/L glucose + 1 ml/L DMSO).Western blotting was performed to detect relative protein quantities of endothelial marker claudin 5 and mesenchymal marker α-smooth muscle actin (α-SMA).TGF-β1 and TGF-β2 mRNA levels were measured by real-time PCR.Vascular endothelial marker VE-cadherin and mesenchymal marker α-SMA were detected by immunofluorescent stain and observed by confocal microscopy.Results Compared with NG,the expression of claudin5 protein in HG (15 or 30 mmol/L) was up-regulated while expression of α-SMA protein was down-regulated (P ＜0.05).Both TGF-β1 and TGF-β2 mRNA levels increased as well (P ＜ 0.05).However,when compared with HG,the claudin 5 levels increased while α-SMA decreased in TGF-β inhibition group.No significant changes were observed in hyperosmotic or solvent control group.Confocal microscopy showed the transformation of cells from a cobblestone-liked shape to a spindle one,and a decreasing expression of VE-cadherin while an increasing α-SMA in HG group (P ＜ 0.05),whereas TGF-β inhibition partly attenuated those changes in both morphological and protein levels.Conclusions High glucose treatment of glomerular endothelial cells results in an increase in the level of TGF-β1 and TGF-β2 mRNA and leads to endothelial-mesenchymal transiton.Inhibition of TGF-β partly prevents this process,indicating that TGF-β plays a crucial role in high-glucose-induced glomerular endothelial-mesenchymal transiton.

Objective To investigate the proteins which were associated with radiosensitivity of nasopharyngeal carcinoma (NPC) cells and could be used to predict the radiosensitivity.Methods A radioresistant subclone cell line CNE-2 (R743) derived from NPC cell line CNE-2 was established.Radiosensitivity and cell cycle characteristics of CNE-2 and CNE-2 ( R743 ) were examined and compared by clonogenic survival assay and flow cytometry.The total proteins from the two cell lines were extracted and separated by two-dimensional gel electrophoresis,and the images were analyzed by Image Master 7.0analysis software.Differentially expressed proteins in the two cell lines were identified through MALDITOF/TOF peptide mass fingerprint and searched in the protein sequence database.The protein expressions were confirmed by RT-PCR and Western blot.Results Totally seven differentially expressed proteins were identified,six of which were upregulated and one downregulated in the radioresistant CNE-2 (R743),compared with those of CNE-2.Three out of the seven,Annexin A2,Tropomyosin 4 and GRP78 were upregulated in the CNE-2 ( R743 ),which were confirmed by Western blot and RT-PCR ( t =24.22,24.20,29.19,P ＜ 0.05).Conclusions Differentially expressed proteins might be involved in different radiosensitivities of nasopharyngeal carcinoma cell lines,among which Annexin A2,Tropomyosin 4 and GRP78 could be the candidate biomarkers for predicting radiosensitivity of nasopharyngeal carcinoma cells.