BACKGROUNDTo investigate the mutation of HBV polymerase gene in chronic hepatitis B patients treated with lamivudine.

METHODSThe restriction-fragment-length-polymorphism (RFLP) assay for HBV DNA sequence determination at the codon 528 and 552 in the HBV polymerase gene associated with lamivudine resistance in vitro. HBV DNA samples extracted from sera of 240 patients were subjected to PCR amplification with primer pairs F2/R2 (552), F3/R2 (528). Each PCR product was digested with Nde I or Nla III.

RESULTSSerum HBV DNA mutation was found in 51/240 patients (38/51M552V, 26/38L528M, 13/51M552I) after therapy for 52 weeks. DNA sequence analysis was performed on samples of 3 patients, and the results were consistent with those of RFLP assay.

CONCLUSIONThe RFLP assay was able to detect the mutation of HBV DNA at codon 552 and 528 which are the principal site of HBV DNA resistant to lamivudine. The specific PCR method for HBV DNA mutation is rapid, simple and specific.

Drug Resistance, Viral ; Gene Products, pol ; genetics ; Hepatitis B virus ; enzymology ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Mutation ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Reverse Transcriptase Inhibitors ; therapeutic use

Brain ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Middle Aged ; Myotonic Dystrophy ; diagnosis ; pathology ; White Matter ; pathology

Objectives To explore the type and subtype distribution of influenza viruses in influenza-like patients and the hemagglutinin(HA)genetic variation of influenza A viruses in Shang- hai and Wuxi during the influenza prevalent season from 2004 to 2006.Methods Throat swabs were collected from the influenza-like patients in the sentinel hospitals and during the outbreaks,and then inoculated into MDCK cells to isolate influenza viruses,which were subsequently identified by direct immunofluorescence(DIF)and reverse transcription-polymerase chain reaction(RT-PCR).HA seg- ments of influenza A viruses were sequenced to analyze the genetic variation of HA.Results One hundred and twenty-six strains of influenza viruses,including 53 H3N2,43 H1N1 and 30 influenza B viruses were isolated from August 2004 to September 2006,and 7 outbreaks.All these outbreaks oc- curred in February or March The pathogens were identified as H1N1 in one outbreak,H3N2 in two outbreaks,B in two outbreaks and mix infections in two outbreaks(1 H1N1 and B,1 H3N2 and B, respectively).By sequencing the HA segment,the H3 and H1 segments were all homologous to the isolates from different countries in the same period.Conclusion H3N2 and H1N1 are the major strains prevalent in Shanghai and Wuxi,which reach the peak from January to March No HA and NA recom- binant strains and new HA and NA subtypes are found in these areas.The variations of H1 and H3 are similar to those found in other countries.

OBJECTIVETo ascertain the genetic characterization and genotype of measles viruses isolated in Shanghai region, in 2005.

METHODSMeasles virus was isolated from throat swab specimens collected from suspected measles cases and 450 bp fragment of C terminus of nucleprotein (N) gene was amplified by RT-PCR. Sequence analysis was conducted to ascertain the genotype and to compare the difference of nucleotide with other measles virus strain published in GenBank.

RESULTS4 measles viruses were isolated from 10 throat swab specimens, and the sequence analysis indicated that they belonged to H1 genotype. The homogeneity of 450 nucleotides in the C terminal of the N gene was at 98%-98.2% as compared to H1 genotype (China93-7). They differed from genotype H2 (China94-1) at 6.4%-6.9% and from genotype A (Edmonston) at 6.7%-6.9%, from measles vaccine (Shanghail91) at 7.6%-8.0%. They differed from the other measles viral strain isolated in China in 1993 - 2005 at 0.2%-3.7%. The variation within 4 isolated measles viruses was at 0.7%-1.3%.

CONCLUSIONIt was H1 genotype measles viruses,which are the native viruses in China that led to the outbreak of measles in Shanghai, in 2005.

China ; epidemiology ; Disease Outbreaks ; Genotype ; Humans ; Measles ; epidemiology ; genetics ; Measles virus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

BACKGROUND: Resting-state functional magnetic resonance imaging (rs-fMRI) is a promising method for the study of brain function. Typically, rs-fMRI is performed on anesthetized animals. Although different functional connectivity (FC) in various anesthetics on whole brain have been studied, few studies have focused on different FC in the aged brain. Here, we measured FC under three commonly used anesthesia methods and analyzed data to determine if the FC in whole brain analysis were similar among groups. METHODS: Twenty-four male aged Wistar rats were randomly divided into three groups (n = 8 in each group). Anesthesia was performed under either isoflurane (ISO), combined ISO + dexmedetomidine (DEX) or α-chloralose (AC) according to the groups. Data of rs-fMRI was analyzed by FC in a voxel-wise way. Differences in the FC maps between the groups were analyzed by one-way analysis of variance and post hoc two-sample t tests. RESULTS: Compared with ISO + DEX anesthesia, ISO anesthesia caused increased FC in posterior brain and decreased FC in the middle brain of the aged rat. AC anesthesia caused global suppression as no increase in FC was observed. CONCLUSION ISO could be used as a substitute for ISO + DEX in rat default mode network studies if the left temporal association cortex is not considered important.
Anesthesia ; Animals ; Brain/diagnostic imaging* ; Brain Mapping ; Isoflurane ; Magnetic Resonance Imaging ; Male ; Rats ; Rats, Wistar

AIM: The aim of this work was to establish a specific and sensitive method to comprehensively investigate and compare chemical constituents of Fuzi-Gancao herb pair (FG), consisting of Aconitum carmichaelii Debeaux (Fuzi, Chinese) and Roast Radix Glycyrrhizae (Glycyrrhiza glabra L., Gancao, in Chinese) and Fuzi alone to explore the underlying interaction mechanism of FG. METHOD: An ultra-fast liquid chromatography-ion trap/time-of-flight mass spectrometry (UFLC/MS-IT-TOF) method using diazepam as internal standard was developed for the identification and semi-quantitative analysis of the phytochemical constituents of Fuzi and FG. Chromatographic separation was achieved on a UFLC column using a gradient program with 40 mmol·L(-1) ammonium acetate and acetonitrile as the mobile phase. RESULTS: Fifty-one of the sixty compounds, including forty-five C19-diterpenoid alkaloids and six C20-diterpenoid alkaloids were tentatively identified in the extracts of Fuzi and FG through accurate mass measurements and fragmentation patterns. Comparing the contents of these alkaloids in these two extracts, it was found that the diester-diterpenoid alkaloids (DDAs) and the alkylolamine-diterpenoid alkaloids (ADAs) were increased, while the monoester-diterpenoid alkaloids (MDAs) were decreased in the extracts of FG. CONCLUSION This work provided comprehensive information for the quality control of Fuzi preparations, and the further investigation on the compatibility mechanisms of FG.
Aconitum ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Interactions ; Drugs, Chinese Herbal ; chemistry ; Glycyrrhiza ; chemistry ; Mass Spectrometry ; methods ; Molecular Structure