Hepatitis B is a serious infectious disease worldwide, and hepatitis B virus (HBV) is the direct cause of this disease. In recent years, as an essential part of its evolutionary process, HBV mutation has been extensively studied domestically and globally. However, the study on the conserved sequences in HBV sequences is still in its infancy. In this study, we applied multiple EM for motif elicitation (MEME) algorithm to discover HBV motif and proposed a new metric, conservative index (CI), to carry out phylogenetic analysis based on HBV sequences. Then, the constructed phylogenetic tree was subjected to reliability assessment. The results demonstrated that the new metric CI combined with the MEME algorithm can effectively help to discover motifs in HBV sequences and construct a phylogenetic tree based on them and to analyze the evolutionary relationship between HBV sequences; in addition, the possible ancestral sequences of samples may be obtained by conservative analysis. The proposed method is valuable for the exploratory study on large HBV sequence data sets.
Computational Biology ; Conserved Sequence ; Evolution, Molecular ; Hepatitis B virus ; genetics ; Humans ; Nucleotide Motifs ; Phylogeny ; Reproducibility of Results

Adult ; Endoscopy ; Epistaxis ; etiology ; surgery ; Female ; Humans ; Middle Aged ; Nasal Cavity ; surgery ; Telangiectasia, Hereditary Hemorrhagic ; complications ; surgery

OBJECTIVE To realize the status and the impact factors of nosocomial infection among Jizheng patients.METHODS To investigate 562 former inpatients suffered from Jizheng in hospital from Jan 2004 to Dec 2005.RESULTS The incidence rates of nosocomial infection that calculated by the patients and the cases were 14.59% and 16.55%,respectively.Infection mostly occurred on the 22nd day in hospital on average.The main infection sites were lower respiratory tract(48.78%),urinary tract(15.85%),and alimentary tract(12.20%).The most risk factor was aging.CONCLUSIONS The high incidence of nosocomial infection is due to various factors.We should take measures respectively aiming at different factors to reduce the occurrence of nosocomial infection.

Hepatitis B is one of the most serious global threats to human health. Phylogenetic analysis of hepatitis B virus (HBV) can reveal the evolutionary relationship between HBV sequences and thus provide a basis for the prediction and treatment of hepatitis B and other aspects. In this study, we performed sequence analyses on the HBV sequences of five clinical HBV samples and the HBV sequences retrieved from the GenBank, EMBL, and DDBJ to construct a phylogenetic tree and analyze sequence structures. The experimental results revealed that the C gene of one cloned sequence had a recombinant structure of HBV B/ C subtype. Moreover, the phylogenetic results proved the existence of a newly found subtype HBV/B6 in Xishuangbanna of Yunnan Province, China. The experimental conclusion represents certain value for phylogenetic studies of HBV in Yunnan ethnic minority groups.
DNA, Recombinant ; genetics ; Genes, Viral ; genetics ; Genotyping Techniques ; Hepatitis B virus ; classification ; genetics ; Humans ; Phylogeny

Objective To prepare controlled release microspheres of vascular endothelial growth factor(VEGF)& calcium alginate and observe their effect on proliferation of human umbilical vein endo- thelial cells(HUVEC)in order to provide theoretical basis for controlled release of VEGF facilitating an- giogenesis of tissue engineering bone.Methods VEGF-calcium alginate microspheres were prepared by using the needle extrusion/external gelation method to investigate physicochemical character and in vitro release of VEGF.According to the different ingredients added into the culture medium,the seconda- ry cultured HUVEC were divided into four groups,ie,control group,microsphere group,VEGF group and VEGF-calcium alginate microsphere group,in which the proliferation of the cultured HUVEC was ob- served with cell counting method,MTT method and flow cytometry.Results The calcium alginate mi- crospheres were revealed as spherical shape and evenly distributed,with mean grain diameter of(560?50)?m,carrying capacity of 0.72 ng/mg and the encapsulation efficiency of 54%.Smooth controlled re- lease in VEGF-alginate microspheres lasted for more than 10 days.Proliferation of the cultured HUVEC was accelerated the most in VEGF group at the beginning but in EGF-calcium alginate microsphere group at midanaphase compared with other groups,with statistical difference(P＜0.05).There was no statis- tical difference upon cell counting,cell activity and time point of cell cycle between control group and mi- crosphere group.Conclusion VEGF-sodium alginate microapheres can continue activity of VEGF,re- lease VEGF for over 10 days and promote proliferation cultured HUVEC for a long time.

Animals ; Carcinogens ; toxicity ; Female ; Lung Neoplasms ; chemically induced ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Sensitivity and Specificity ; Urethane ; toxicity

Objective To explore the pathogenic bacteria species of the patients with lactation acute mammitis and their sensitivities to antibiotics in order to provide a guideline in choosing antibiotics reasonably. Methods Four hundred and thirty-three samples from diseased breast(include milk,puncture fluid and secretion in ulcerateing skin)from 310 patients with lactation acute mammitis,who were treated by the center of mammary gland of Maternal and Child Health Care of Haidian district in BeiJing from Jan. to Aug. in 2012. Statistics analysis was made to analysis the pathogenic bacteria species and the characteristics of drug sensitivity. Results (1)Of the 433 samples,407 strains of pathogens were picked out,which consisted of 215 strains of Staphylococcus aureus(SAU),43 strains of methicillin-resistant staphylococcus aureus( MRSA),43 strains of coagulase negative staphylococcus(CNS),42 strains of Methicillin-resistant staphylococcus epidermis (MRSE),22 strains of Staphylococcus epidermidis(SE),12 strains of Methicillin-resistant coagulase negative staphylococci(MRScon)and 8 strains of Alpha hemolytic streptococcus.(2)There were 237 strains of G +bacteria,and the drug sensitive test showed that they were all sensitive to vancomycin(100% ),91. 8% were sensitive to penicillin,57. 0% to clindamycin( CLDM))and 69. 0% to erythrocin. Meanwhile,94. 0% were sensitive to levofloxacin,70. 0% to cephalosporins,and 80. 0% to gentamicin. In addition,there were 43 MRSA, and drug sensitivity test showed that 95. 0% were sensitive to levofloxacin,96. 0% to gentamicin,97. 0% to macrodantin,and 92. 0% to cotrimoxazole. Meanwhile,it's drug resistance rate to erythrocin was 13. 0% and 15. 0% to clindamycin( CLDM). MRSA was completely resistant to Lactam antibiotics( 100% ) such as penicillin and Cephalosporins,95. 0% to levofloxacin,96. 0% to gentamicin,97. 0% to furadantin and 92. 0% to bactrim. Conclusion The predominant pathogenic bacteria of lactation acute mammitis include SAU,MRSA, MRSA,CNS,MRSE,SE and they are all the G + bacteria. They are showed with a high rate of drug resistance, especially for the sensitive drugs which had been proved in the past. Due attention should be paid to germiculture from diseased breast and drug sensitive test as soon as possible,and to provide a guideline in choosing antibiotics reasonably.

Objective Streptococcus pneumoniae can form biofilms .The aim of this study was to investigate the biofilm forma-tion of Streptococcus pneumoniae and the relationship with antibiotic resistance of penicillin etc . Methods A total of 147 clinical iso-lates of Streptococcus pneumoniae were collected from 7 teaching hospitals in Nanjing from 2010 to 2012.Minimal inhibitory concentration (MIC) of penicillin, erythromycin, cefuroxime and ceftriaxone were determined by agar dilution method .Streptococcus pneumoniae with various penicillin MIC was selected randomly as follow:MIC≤0.065μg/mL, 0.5μg/mL, 2μg/mL and≥4μg/mL, which was incuba-ted to form biofilms in 96-well plates and 24-well plates for 24 hours.The A values at 570 nm was measured and the biofilm was observed through confocal laser scanning microscope ( CLSM) . Results The biofilm semi-quantitative detection and CLSM both displayed that all strains formed biofilms.The A value of the group which penicillin MIC was ≤0.065μg/mL (0.228 ±0.063) was higher than the 0.5μg/mL group (0.200 ±0.061) and the≥4μg/mL group (0.186 ±0.050) (P<0.05) , and there was no difference among the groups which penicillin MIC were 0.5μg/mL, 2μg/mL and≥4μg/mL, respectively (P>0.05).The A value of the group which erythromycin MIC was ≤0.5μg/mL (0.211 ±0.068) was higher than the ≥4μg/mL group (0.201 ±0.052) (P>0.05).The A value of the group sensitive to cefuroxime (0.216 ±0.062) was higher than the group resistant to cefuroxime (0.196 ±0.054) (P<0.05). Conclusion Streptococcus pneumoniae can form biofilms .Streptococcus pneumoniae with high antibiotics MIC has a trend of weakened biofilm formation .

Objective To explore the effects of theophylline plus salmeterol/fluticasone propionate combination product (SFC) on clinical control,lung function and airway inflammation in asthmatics.Methods A total of 146 asthmatics received 200 mg theophylline plus 50/250 μg SFC twice daily for 24 weeks.The level of asthma control was assessed by the asthma control test.Testing of lung function and inflammatory markers in induced sputum were performed.And 142 asthmatics received 50/250 μg SFC twice daily for 24 weeks as control.Results Asthma was completely controlled in 61 and 59 in the theophylline plus SFC and SFC groups respectively after a 24-week treatment period (P ＞ 0.05).Theophylline plus SFC improved the FEF25％ 75％ value,indicating small airway function,to a greater extent than SFC [(66.7 ± 18.2) ％ & (56.6 ± 17.4) ％,P ＜ 0.01].Percentage of eosinophil and concentration of eosinophil cationic protein in induced sputum were significantly lower in the theophylline plus SFC group than those in the SFC group [(4.1 ±2.3)％ vs.(6.2±2.7)％ & (63.9±39.4) vs.(90.3 ±46.2) μg/Lrespeetively] (all P ＜ 0.01).Conclusion The therapy of theophylline plus SFC may provide greater improvements in small airway function and airway inflammation.

A geldanamycin (GDM) producing strain, Streptomyces hygroscopicus 17997, was isolated from Yunnan China soil by our institute researchers. GDM is an ansamycin antibiotic, which has the ability to bind with Hsp90 (Heat Shock Protein 90) and alter its function. Hsp90 is a chaperone protein involved in the regulation of the cell cycle, cell growth, cell survival, apoptosis, and oncogenesis. So it plays a key role in regulating the physiology of cells exposed to environmental stress and in maintaining the malignant phenotype of tumor cells. As an inhibitor of Hsp90, GDM possesses potent antitumor and antivirus bioactivity, but the hypato-toxicity and poor solubility in water limits its clinical use. Two GDM derivatives, 17-(Allylamino)-17-demethoxygeldanamycin (17-AAG) and 17-dimethylamino-ethylamino-17-demethoxygeldanamycin (17-DMAG), both showing lesser hepato-toxicity, are now in Phase II and Phase I clinic trials. In order to accomplish the structure modification of GDM by genetic means, an attempt to obtain the biosynthetic gene cluster of GDM from S. hygroscopicus 17997 was made. In this study, a pair of primers was designed according to a conserved sequence of one of possible post-PKS (polyketides synthase) modification genes, the carbamoyltransferase (CT) gene (gdmN) in GDM biosynthesis. The 732 bp PCR product was obtained from the S. hygroscopicus 17997 genomic DNA. Through the colony-PCR Binary Search Method, using the CT gene primers, six positive cosmid clones, CT1-6, were identified from the S. hygroscopicus 17997 cosmid genomic library. The CT gene containing fragments were verified and localized by Southern blot. The CT-4 positive cosmid was then sub-cloned and sequenced. Approximately 28.356kb of foreign gene sequence from CT-4 cosmid and by further PCR extension reaction was obtained. Based on BLAST analysis, this sequence contains 13 possible ORFs and their deduced functions are believed to be involved in GDM production. The ORF1 encoding products show homology (87%) with incomplete sixth module and complete seventh module of PKS, gdmA3, in S. hygroscopicus NRRL 3602. The ORF2-13 gene products are similar to gdmF(9 5%), gdmM(8 8%), gdmN (92%), gdmH (92%), I (93%), J (90%), K (93%), G (96%), gdmO (91%), gdmP (93%) and two transcription regulation genes gdmRI (83%) and gdmRII (90%). The obtained possible GDM biosynthetic gene cluster in S. hygroscopicus 17997 will facilitate the further functional analysis of the genes and to modify the structure of GDM through combinatorial biosynthesis.
Anti-Bacterial Agents ; metabolism ; Benzoquinones ; metabolism ; Carboxyl and Carbamoyl Transferases ; genetics ; Chromosome Mapping ; Cloning, Molecular ; DNA Primers ; genetics ; Lactams, Macrocyclic ; metabolism ; Multigene Family ; Streptomyces ; genetics ; metabolism