Little is known about the non-structural protein 6(NSP6)of rotavirus.This report describes expression of the NSP6 of a group A human rotavirus strain TB-Chen in bacteria,and its immunological properties and cellular distribution.The results showed that the recombinant NSP6(rNSP6)was expressed in high efficiency without any other proteins fused(possesses about 34.2% of total bacterial proteins).rNSP6 elicited mono-specific antibodies in immunized guinea pigs and the antibodies could react with the rNSP6 itself and the viral NSP6 proteins synthesized in SA11-or Wa-infected MA104 cells in Western blot and immunofluorescence assay.The NSP6 distributed evenly in the cytoplasm mainly around the nucleus of virus-infected cells,no viroplasm-like gathering observed;The top amount of NSP6 synthesized in SA11-infected cells or Wa-infected cells could be detected at 12h after infection.This is the first report about the high expression of entire NSP6(without any other proteins fused)in prokaryotic expression system and detection of NSP6 synthesis in virus infected cells by immunofluorescence assay.The results are important to understand the structure,biological properties and further application of the NSP6.

By means of genetic cloning and recombinant techniques, full genome cDNA sequences of rotavirus strain TB-Chen were isolated from an infantile hospitalized with acute gastroenteritis. Nucleotide sequences analyses showed that the full genome of strain TB-Chen contains 18613 nucleotides, encoding 5791 amino acids. Genotyping results showed that the strain TB-Chen belongs to genotype G2P[4]/NSp4[A]. This is the first report on a full genome of Group A rotavirus in China, and has important significance for deep understanding structure and functions of rotaviruses and developing rotavirus vaccines.

Objective To evaluate the efficacy and safety of intravesical instillation of sodium hya- luronate in relieving the symptoms of refractory non-bacterial cystitis.Methods Totally,20 patients(1 man and 19 women;mean age,47 years;age range,22-68 years)with refractory non-bacterial cystitis were included.Their disease course ranged from 1 month to 30 years.Of them,16 patients had the disease for more than 1 year.Instillation of sodium hyaluronate(50 ml solution of 40 mg HA)was performed intravesically in them for a total of 12 weeks(once a week for the first 4 weeks and once every 4 weeks for the succeeding 8 weeks).Follow-up visit ended at 16 weeks,when the patients filled in the symptom assessment scores(VAS on pain,frequency and urgency)according to the subjective evaluation.Therapeutic results were classified as(1)total response:symptom disappearance or relief by at least 90% of the symptom scores;(2)partial response:symptom relief by 50%-89%;(3)mild response:symptom relief by 49% or less;(4)no re- sponse:no improvement in symptom scores.Results Nineteen patients completed the 16-week follow-up with only 1 withdrawing.The average general symptom score decreased from 22.32?5.53 to 9.47?5.88 (P＜0.001);frequency score from 8.21?1.75 to 3.89v2.31(P＜0.001);urgency score from 7.47?2.14 to 3.37?2.14(P＜0.001);pain score from 6.63?3.47 to 2.21?2.74(P＜0.001).After 16 weeks,overall positive response rate(total+partial response)was 53%(10/19).No serious adverse effect occurred except for urethral irritation(25%,5/20),worsening of distending pain of the bladder(5%,1/20) and vertigo(15%,3/20)caused by the procedure itself.Conclusions For patients suffering from refrac- tory non-bacterial cystitis,intravesical instillations of sodium hyaluronate can significantly relieve the symp- toms and is safe and of good patient compliance.

Objective Discuss the anesthetic effect ofremifentanil and fentanyl on interventional surgery for children with congenital heart disease.Methods 80 patients with congenital heart disease were selected,they were divided into two groups randomly.The control group (40 cases) was given fentanyl anesthesia.The observation group (40 cases) was given remifentanil anesthesia.Observe and record SDP,DBP,HR at different time of anesthesia,extubation time,recovery time and dosage of anesthesia and adverse reaction during anesthesia to evaluate the anesthetic effect of remifentanil and fentanyl on interventional surgery for children with congenital heart disease.Results Compared with the basic value before induction of anesthesia,the SDP,DBP of two groups were decreased after induction (P ＜ 0.05).There was no significant difference on HR level between two groups before and after induction.The extubation time,recovery time of observation group was shorter than that of the control group.However,the amount of anesthesia drug in the observation group was higher than that of the control group (P ＜ 0.05).During anesthesia,there was no significant difference on adverse reactions between two groups.Conclusion The anesthetic effect of remifentanil on interventional surgery for children with congenital heart disease is good,and it has little effect on cardiac function with early extubation and less side effect.

The non-covalent interactions between 18-crown-6 (18c6) and 20 common types of protonated amino acids were explored by electrospray ionization mass spectrometry (ESI-MS).The mass spectra showed the formation of 1:1 stoichiometric non-covalent complexes between 18c6 and amino acids.The calibration curves and linear equations for the complexes of L-Phe,L-Tyr,L-Lys and L-Asp with 18c6 were established by mass spectrometric titration and used as reference values for competitive ESI-MS.Through competitive equilibrium,the binding constants for the complexes of 18c6 with other L-amino acids and their D-isomers were derived.It was found,as a general trend,lgKa for the complexes of 18c6 with the basic amino acid and the amino acid with alkyl side chain were larger than other complexes,and among the amino acids with alkyl side chain,Gly and Ala exhibited greater 18c6 binding affinities.As for Ser and Thr,the intramolecular hydrogen bond between the nitrogen atom from terminal NH2and the oxygen atom from carboxyl may impede their protonated amino-group to attack the 18c6.Furthermore,Gln and Asn exhibited lower 18c6 binding affinities probably due to effects of electron-withdrawing group of acylamide.Finally,the chiral selectivity of 18c6 for 19 L-,or D-amino acids was measured by ESI-MS,indicating 18c6 could only recognize some neutral amino acid isomers.

Drug adulteration and contamination are serious threats to human health therefore,their accurate monitoring is very important.Allopurinol(Alp)and theophylline(Thp)are commonly used drugs for the treatment of gout and bronchitis,while their isomers hypoxanthine(Hyt)and theobromine(Thm)have no effect and affect the efficacy of the drug.In this work,the drug isomers of Alp/Hyt and Thp/Thm are simply mixed with α-,β-,y-cyclodextrin(CD)and metal ions and separated using trapped ion mobility spectrometry-mass spectrometry(TIMS-MS).TIMS-MS results showed that Alp/Hyt and Thp/Thm iso-mers could interact with CD and metal ions and form corresponding binary or ternary complexes to achieve their TIMS separation.Different metal ions and CDs showed different separation effect for the isomers,among which Alp and Hyt could be successfully distinguished from the complexes of[Alp/Hyt+y-CD+Cu-H]+with separation resolution(Rp-p)of 1.51;whereas Thp and Thm could be baseline separated by[Thp/Thm+y-CD+Ca-H]+with Rp-p of 1.96.Besides,chemical calculations revealed that the complexes were in the inclusion forms,and microscopic interactions were somewhat different,making their mobility separation.Moreover,relative and absolute quantification was investigated with an internal standard to determine the precise isomers content,and good linearity(R2＞0.99)was ob-tained.Finally,the method was applied for the adulteration detection where different drugs and urine were analyzed.In addition,due to the advantages of fast speed,simple operation,high sensitivity,and no chromatographic separation required,the proposed method provides an effective strategy for the drug adulteration detection of isomers.

Objective To study the outcomes of AIDS intervention programs and to provide scientific evidence for developing pertinent strategy on intervention among men who have sex with men (MSM).Methods MSM were recruited through snowballing and investigated by face to face interview in 2006 and 2007 respectively.SPSS 12.0 was used to compare the change of cognition about AIDS,sexual behavior,prevalence rates of HIV and Syphilis before and after the intervention program among the population under study.Results The cognition about AIDS among MSM was obviously improved after the intervention with the rate increased from 74.3% to 82.4% (P=0.01).The rate of last time condom use among MSM increased from 56.4% to 65.5% (P=0.00).The rate of consistent condom use during six months among MSM increased from 31.8% to 41.9% (P=0.00).The rates of both condom use during commercial sex with men and with women did not change much among MSM.The prevalence rates of HIV in 2006 and 2007 were 10.4% and 10.8% and of syphilis in 2006 and 2007 were 9.3% and 7.3% respectively which were not significantly different between before and after the intervention.Conclusion The cognition about AIDS among MSM was improved obviously.However,the rate of consistent condom use was still low,reflecting the segregation phenomenon between their behavior and cognition.All our findings implied that it was crucial to carry out AIDS prevention and control programs.

Strain TB-Chen is a group A rotavirus (RV) isolated from a Chinese infant suffering from gastroenteritis in hospital. The NSP5 and NSP6 of strain TB-Chen are encoded by the 10th gene segment (816bp in whole length) of the viral genome. The results obtained in this study showed that the NSP5 was encoded in the first open-reading-frame (ORF) of the gene segment (from 22bp to 624bp), and NSP6 was encoded in the second ORF (from 80bp to 355bp). The NSP5 protein consisted of 200 amino acid residues with a putative molecular mass of 21.9 kD, and a putative isoelectric point of 7.86. The NSP6 protein consisted of 92 amino acids with a putative molecular mass of 11 kD, and a putative isoelectric point of 9.65. This study further analyzed phylogenetic relationship of the NSP5/NSP6 ORF nucleotide sequence. The results showed that the NSP5s of group A rotavirus could be at least classified into 7 genotypes (H1-H7), the NSP6s could be at least classified into 8 genotypes (hl-h8); the genotypes of the NSP5 and NSP6 derived from strain TB-Chen was classified as H2 and h2. This was the first report on the genotype classification of the NSP6 of group A RVs, and it was proposed English letter "h" to represent genotype of the NSP6, e. g. strain 69M classified as H7h7, strains Wa and KU classified as H1h8.
Evolution, Molecular ; Gastroenteritis ; virology ; Genotype ; Humans ; Open Reading Frames ; genetics ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; classification ; genetics ; Viral Nonstructural Proteins ; genetics

Differences of protonated and lithiated leucine-enkephalin(LE) were investigated by hydrogen deuterium exchange-mass spectrometry(HDX-MS) combined with quantum chemistry calculation. The results revealed that the protonated ions possessed very high product yield with all hydrogen atoms being exchanged, while the reaction of lithiated LE stopped after exchanging five hydrogen atoms in the same experimental conditions. The different HDX behaviours probably indicated their conformational differences. To further clarify the experimental results, the most stable conformations of protonated and lithiated leucine-enkephalin were calculated by density functional theory. It was found that terminal amino group was the most thermodynamically stable protonation site,while Li+in coordination of four carbonyl oxygen atoms formed the most favourable lithiated LE. The reaction field reduction of lithium LE was probably due to the less acidity of hydrogen atoms and the increasing rigid conformation change induced by lithium ion.

OBJECTIVETo perform variation and phylogenetics analysis on the SARS-CoV genome sequence (PUMC01) isolated in the Peking Union Medical College Hospital.

METHODSThe cDNA library of SARS-CoV (PUMC01 isolate) was constructed by means of random-priming strategy. Random selected plasmid was sequenced and the genome sequence of SARS-CoV-PUMC01 was assembled by conventional methods (The Genebank Accession No. of SARS-CoV-PUMC01 is AY350750). The variation and phylogenetics analysis were performed by comparing the PUMC01 sequence with other SARS-CoV isolates.

RESULTSTen variation sites were found by comparing PUMC01 isolate with Tor2 and Urbani isolates. In phylogenetic analysis of 18 SARS-CoV isolates, two classes were observed and there is different differential time between these two classes and the different isolates in each class.

CONCLUSIONSThe evidence of phylogenetic analysis of different SARS-CoV isolates from different region is instructive for understanding the clinical relations between the different isolates and the transmission chain of SARS-CoV.

Amino Acid Sequence ; Base Sequence ; China ; DNA, Viral ; genetics ; Genetic Variation ; Genome, Viral ; Molecular Sequence Data ; Phylogeny ; SARS Virus ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Viral Proteins ; genetics