Objective To investigate the infections caused by Staphylococcus aureus carrying Panton-Valentine leukocidin(PVL)genes.Methods 26 isolates of Staphylococcus aureus carrying Panton- Valentine leukocidin(PVL)genes were determined by multiplex PCR.Multilocus sequence typing(MLST) was used to determine the STs of the isolates.The genotypes of SCCmec were also determined by another multiplex PCR in the isolates of methicillin-resistant Staphylococcus aureus(MRSA).Results Among 26 isolates,there were 6 isolates of ST88 MRSA,7 isolates of ST88 methicillin-susceptible Staphylococcus aureus (MSSA),5 isolates of ST239 MRSA,5 isolates of ST398 MRSA,1 isolate of ST25 MRSA,1 isolate of ST30 MRSA and 1 isolate of ST59 MRSA.20 isolates were hospital-acquired(HA)which mainly caused pulmonary infection and post-operative pyogenic infection.6 isolates were community-acquired(CA)which mainly caused soft tissue necrosis.Among 19 isolates of MRSA,ST88-SCCmec Ⅲ A,ST239-SCCmec Ⅲ,ST398- SCCmec Ⅳ and ST398-SCCmec Ⅲ were main types.26 isolates were isolated from 14 wards.ST88-SCCmec Ⅲ A-MRSA caused clone spread in maternity department in our hospital.Conclusion ST88,ST239 and ST 398 are main STs in Staphylococcus aureus carrying PVL in our hospital.The isolates not only cause nosocomial infections but also cause community infection.

OBJECTIVETo find the development rules of microbial community in rhizoma sphere of the cultivated Atractylodes lancea.

METHODTotal bacteria, fungi and actinomyces were counted by CFU x g(-1) though dilution plate method. And genomic DNA of microbes were extracted and amplified by primers of E. coli's 27f and 1492r to get the 16S rDNA, then the restriction endonuclease Hinf was used to digest the 16S rDNA.

RESULTTotal bacteria, fungi and actinomyces in 2-year old soil were lower than in 1-year old soil, they decreased 46. 14%, 49. 25%, 31.88% respectively and made the ratio of themselves changed. At the same time, all the 8 soil samples got fine 16S rDNA bands, which were about 1500 bp. And the main bands of most of the samples were found at 1000 bp, but the weak bands of each were different although most bands in the same year samples were more similar than in different year ones.

CONCLUSIONIt is indicated that the change of soil microbial community may has some relation to the continous cropping barrier of A. lancea.

Actinomyces ; genetics ; isolation & purification ; Atractylodes ; growth & development ; Bacteria ; genetics ; isolation & purification ; Biodiversity ; Colony Count, Microbial ; DNA, Ribosomal ; genetics ; Fungi ; genetics ; isolation & purification ; Plants, Medicinal ; growth & development ; RNA, Ribosomal, 16S ; genetics ; Rhizome ; growth & development ; Soil Microbiology

OBJECTIVETo investigate molecular epidemiologic features of rotaviruses circulating in Shanghai, China.

METHODSStool samples were collected from 1230 hospitalized children with community-acquired and nosocomially acquired diarrhea in Children's Hospital Affiliated to Fudan University between November 1, 1999 and December 31, 2001. Polyacrylamide gel electrophoresis (PAGE) was used to detect rotavirus genomic RNA and identify electropherotypes of group A rotavirus RNAs. Reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify full length VP7 gene and dot blot hybridization was performed to identify rotavirus G serotypes using digoxigenin-labelled variable regions from VP7 genes as probes. These probes were amplified by PCR from recombinant plasmids containing full length G1, G2, G3 and G4 VP7 genes from rotavirus field strains detected in Beijing and digoxigenin labelled dUTP was integrated into the PCR products. The Kruskal-Wallis analysis of variance was employed to analyze whether there were significant differences in variables.

RESULTSOut of 1230 samples investigated, 493 (40.1%) were group A rotavirus gene positive by PAGE, among which 397 (80.5%) showed long electropherotypes, 55 (11.2%) showed short electropherotypes, 18 (3.7%) showed mixed electropherotypes which suggested that the children were co-infected by rotaviruses with different electropherotypes, 23 (4.7%) were non-typable because of degradation of some of the genomic RNA fragments. No group B or group C rotavirus was found. RT-PCRs were performed for 328 fecal specimens containing sufficient rotavirus RNAs and VP7 gene products were obtained from 254 (77.4%) samples. Dot blot hybridization showed serotype G1 accounted for 55.5% (141) of these samples, serotype G3 accounted for 27.6% (70), serotype G2 accounted for 9.4% (24), co-infection by 2 rotaviruses with different G types accounted for 6.3% (16), only 1 G4 was detected and 2 were non-typable. The genomic RNA patterns of all G2 strains were short and those of G1, G3 and G4 strains were long. There were no statistically significant differences for age distribution and clinical manifestations among those infants and children infected by rotaviruses with different G serotypes.

CONCLUSIONGroup A rotavirus is the major pathogen for diarrhea in infants and children in Shanghai during the period of Nov. 1999 to Dec. 2001. Rotaviruses with long electropherotype were dominant during these years. Serotypes G1 to G3 constituted 98.8% of all 254 strains tested, and G1 was the most common serotype followed by G3 and G2, whereas serotype G4 was seldom found. Some of the children were co-infected by rotaviruses with different G serotypes. Clinical manifestations were not related to the infecting rotavirus with different G serotypes.

Age Factors ; Antigens, Viral ; Capsid Proteins ; genetics ; metabolism ; Child, Preschool ; China ; epidemiology ; Data Collection ; Dysentery ; epidemiology ; etiology ; Electrophoresis, Polyacrylamide Gel ; Feces ; virology ; Female ; Humans ; Infant ; Male ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; classification ; genetics ; Rotavirus Infections ; complications ; epidemiology ; virology ; Serotyping

Hot-melt extrusion was applied to prepare mesoporous silica/ethylcellulose mini-matrix for sustained release, and fenofibrate was used as a model drug, ethylcellulose and xanthan gum were chosen as sustained-release agent and releasing moderator, respectively. This novel matrix obtained the controlled release ability by combining mesoporous silica drug delivery system and hot-melt extrusion technology. And mesoporous silica particle (SBA-15) was chosen as drug carrier to increase the dissolution rate of fenofibrate in this martix. Scanning electron microscope, transmission electron microscope, small angle X-ray powder diffraction and N2 adsorption-desorption were introduced to determine the particle morphology, particle size and pore structure of the synthesized SBA-15. The results showed that SBA-15 had a very high Brunauer-Emmett-Teller specific surface area, a narrow pore size distribution, large pore volume and a ordered two-dimensional hexagonal structure of p6mm symmetry. Differential scanning calorimetry and X-ray powder diffraction results demonstrated that fenofibrate dispersed in an amorphous state inside the pores of the mesoporous silica which contributed to the improvement in the dissolution rate. The drug release of mini-matrices was influenced by ethylcellulose viscosity grades and xanthan gum concentration, which increased with the increasing of xanthan gum concentration and decreasing of ethylcellulose viscosity. Mini-matrix containing 22% xanthan gum exhibited a good sustained release performance, and the drug release behavior followed the first-order kinetics.
Adsorption ; Calorimetry, Differential Scanning ; Cellulose ; analogs & derivatives ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Particle Size ; Porosity ; Powder Diffraction ; Powders ; Silicon Dioxide ; Solubility ; X-Ray Diffraction

OBJECTIVEIdiopathic nephrotic syndrome (INS) is a common glomerular disease. The pathogenesis of the disease remains unclear. Recent studies indicate that transforming growth factor beta (TGF beta) is the main cytokine involved in glomerular disease. It plays an important role in the development of INS and in occurrence of glomerulosclerosis. The present study aimed to study changes and significance of TGF beta in children with idiopathic nephrotic syndrome (INS).

METHODSTotally 35 cases with INS (13 males, 22 females) were studied. The age of onset was between 2 years and 1 months and 14 years with an average of 8 years and 3 months. The active stage group had 35 cases and the remission stage groups had 25 cases. The cases in active stage group had first onset of the disease with obvious clinical symptoms and abnormal laboratory findings without use of corticosteroids. The cases in remission stage group were asymptomatic without abnormal laboratory findings. Protein in urine was negative over 4 weeks after oral administration of prednisone for 8 weeks. Twenty five cases were steroid responsive and 10 cases were steroid non-responsive among the 35 cases. Thirty healthy young children were enrolled as control. TGF beta was detected by ELISA in peripheral blood mononuclear cell (PBMC) culture medium. The TGF beta mRNA gene expression was measured by in situ PCR in PBMC.

RESULTS(1) Concentration of TGF beta(247 +/- 26) ng/L and TGF beta mRNA expression (0.57 +/- 0.18) in active stage of simple type or nephritis type INS were higher than those of remission stage and control (P < 0.01). Concentration of TGF beta[(125 +/- 16) ng/L] and TGF beta mRNA expression (0.30 +/- 0.12) in remission stage were higher than that of control (P < 0.05). (2) The level of TGF beta protein in nephritis type [(275 +/- 26) ng/L] was significantly higher than that in simple type [(220 +/- 18) ng/L] in active stage INS (t = 6.45, P < 0.01). No significant difference in TGF beta mRNA expression was found between the nephritis type (0.58 +/- 0.15) and simple type (0.55 +/- 0.16) in active stage INS, either (P > 0.05). But these two types were different from the control (P < 0.01). (3) Concentration of TGF beta and TGF beta mRNA expression after therapy was clearly lower than that before therapy in steroid responsive group (P < 0.01). Whereas no significant change was seen in steroid non-responsive group. Both indicators were higher in steroid non-responsive group than in steroid responsive group whether before or after therapy.

CONCLUSIONTGF beta may play an important role in the mechanism of INS and its level in PBMC can be used as an immunological indicator for the illness state, therefore, determination of TGF beta level and mRNA may be of some clinical significance.

Adolescent ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Nephrotic Syndrome ; blood ; drug therapy ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; genetics ; metabolism

Objective To investigate the genetic polymorphism of mec Ⅰ in the clinical isolates of methicillin-resistant Staphylococcus anreus(MRSA).Methods 40 isolates(MRSA)carrying mecA gene were selected randomly from the clinical isolates of Staphylococcus anreus from Jan,2005 to Aug,2006 in our hospital.The mec Ⅰ gene was detected by PCR followed with sequencing.Staphylococcal cassette chromosome mec(SCCmec)in MRSA were detected by multiplex-PCR.Agar dilution method was used for determining the MICs of oxacillin against MRSA.Results 35 of 40(87.5%)MRSA carried mec Ⅰ gene.All isolates carrying mec Ⅰ gene have mecI 202C→T substitution,which resulted in Gln at 68 aminophenol position replaced by stop condon.32 isolates carried single point mutation.3 isolates carried double-point mutation,including additonal A at 3 positon,A→C at 41 position and C→T at 142 position beside C→T at 202 position,respectively.Among 35 isolates carrying mec Ⅰ gene,there were 27 isolates of SCCmec Ⅲ, 7 isolates of SCCmec Ⅲ A and 1 isolate of SCCmec Ⅱ.Among 5 isolates with deletion of mec Ⅰ gene,there were 3 isolates of SCCmecⅣ,1 isolate of SCCmec Ⅰ and 1 isolate of non-known SCCmec tpye.The MICs of oxacillin were 256-512 ?g/ml,≥512 ?g/ml and 8-256 ?g/ml in 31 isolates with single point mutation at 202 position in mec Ⅰ gene,3 isolates with double-point mutation in mecI gene and 5 isolates with deletion of mec Ⅰ gene,respectively.1 isolate with single point mutation in mec Ⅰ gene had contrary result(MIC

Objective To explore the cognitive level and demand of chronic disease prevention and treatment integration in the County hospitals and primary health care institutions. Methods A stratified cluster sampling method was used to investigate the medical staff of five county-level hospitals and 39 community health service centers in Liandu District, Yunhe County and Jingning County, and qualitative interviews and on-site questionnaire survey were carried out among 573 medical staff from August to October in 2016. Results A total of 252 medical personnel at the county level or above, accounting for 43.98％, and 321 medical personnel in primary health care institutions, accounting for 56.04％. And 96.86％ of the medical staff thought it is necessary to integrate medical treatment and prevention. Only 32.98％ think that the local medical and anti-integration were the real ones and only 36.13％ have contacted the"top five prevention and control offices" at the county level. Two-way referral of key chronic patients and promotion of grassroots promotion of appropriate technology were better. And 77.38％ of medical staff at medical institutions above the county level and 75.70％ of medical personnel of primary medical institutions participated in the two-way referral work, with 66.67％ of county level medical staff of above medical institutions and 93.46％ medical staffs of primary medical institutions participated in the promotion of grassroots workplaces for appropriate technologies. And 82.72％ of the medical staff held or participated in appropriate technical training courses for chronic diseases within one year, but the proportion of holding or participating in ≥3 times was only 24.08％. Conclusion The work that county level five platform to promote chronic disease prevention and control of chronic disease prevention and treatment of medical integration still need to be strengthened. We should use the appropriate training mode to improve comprehensive prevention and treatment of chronic diseases among primary medical staff.

OBJECTIVETo increase the sensitivity and specificity of conventional gene diagnosis of facioscapulohumeral muscular dystrophy 1A(FSHD1A) by analyzing the distribution of translocation between chromosomes 4q35 and 10q26 in suspected FSHD cases.

METHODSThe Bgl II- Bln I dosage test was performed to detect translocation between chromosomes 4q35 and 10q26 in 7 cases of presymptomatic FSHD patients showing positive result in gene diagnosis and 5 cases of sporadic FSHD patients showing negative result in gene diagnosis. DNA samples were digested with Bgl II and Bln I, followed by agrose gel electrophoresis. Probe p13E-11 was labeled with alpha-(32) P dCTP, followed by Southern hybridization. Then the ratio between the chromosomes 4 and 10 derived signal intensities was judged and hence was made known whether there was interchromosomal translocation between chromosomes 4 and 10.

RESULTSThe Bgl II-Bln I dosage test revealed a translocation from chromosome 4q35 to 10q26 in one presymptomatic FSHD patient, thus indicating the result of gene diagnosis for her might be false positive. There was one translocation from chromosome 10q26 to 4q35 detected in one sporadic FSHD patient, indicating the result of gene diagnosis for her might be false negative. There were no translocations between chromosomes 4 and 10 in the other 10 cases.

CONCLUSIONThe Bgl II-Bln I dosage test can detect the translocation between chromosomes 4q35 and 10q26. It can improve the accuracy of the conventional method for gene diagnosis of FSHD1A.

Adolescent ; Adult ; Bacterial Proteins ; pharmacology ; Child ; Child, Preschool ; Deoxyribonucleases, Type II Site-Specific ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Muscular Dystrophy, Facioscapulohumeral ; diagnosis ; genetics ; Nuclear Proteins ; Proteins ; genetics ; Translocation, Genetic

OBJECTIVETo screen and detect the female carriers from the DMD/BMD family members for prenatal or preimplantation genetic diagnosis.

METHODSFor the detection of DMD/BMD carriers from 27 family members in 4 families, PCR to five microsatellite markers(located in 5' terminus and intron 44, 45, 49, 50) and analysis of the short tandem repeat(STR) sequence polymorphism with the use of genescan were implemented.

RESULTSSix of the 17 female members were obligate DMD gene carriers according to the haplotype analysis of the results of the genescan, which conformed with the pedigree analysis. Besides, the authors detected five carriers and five normal females in these families with the use of the haplotype analysis only. The most polymorphic locus was STR 49, and the least was STR 50.

CONCLUSIONThe STR haploid linkage analysis using (CA)n repeats within the human dystrophin gene is a rapid,accurate, objective method and is well suited for routine use in clinical laboratories engaged in DMD/BMD linkage analysis for the detection of carrier.

Female ; Genetic Carrier Screening ; Humans ; Male ; Microsatellite Repeats ; Muscular Dystrophy, Duchenne ; genetics ; Polymerase Chain Reaction ; Tandem Repeat Sequences

The United Nations Environment Program estimates that approximately 20% of agricultural land and 50% of cropland in the world is salt-stressed. The gene NHX (Na+/H+ exchanger) encodes functional protein that catalyzes the countertransport of Na+ and H+ across membranes and may play an important role in plant salt tolerance. To clone the NHX from the wild plant Populus euphratica collected in Tarim basin and Xinjiang Wujiaqu district into a T-vector, designed primer was used to amplify 1kb NHX cDNA fragment with RT-PCR. Total RNA was extracted from Populus euphratica tissue (plant tissue was collected from Tarim basin and Xinjiang Wujiaqu district and stored in liquid nitrogen) according to the Plant RNA Mini Kits of Omega. First cDNAs were synthesized from 1 microg total RNA of Populus euphratica seedling. A pair of primers were used to perform RT-PCR. The amplified DNA fragment was purified and cloned into pMD18-T vector. However, 1kb and 2.3kb fragment were obtained from Tarim basin and Xinjiang Wujiaqu district and named as PtNHX and PwNHX, respectively. Sequence analysis reveals that the cloned PtNHX fragment of Populus euphratica contains partial NHX coding region with 98%, 86%, 84% and 80% identity comparing with Atriplex gemelini, Suaeda maritima, Arabidopsis thaliana and Oryza sativa, respectively. This analysis suggests that NHX gene would be highly conserved in terms of evolution in plant; and it also suggests that the NHX gene of Populus euphratica also would have the similarity with that of Arabidopsis. It may be of great importance in improvement of the plant salt tolerance and breed of crop. At the same time, sequence analysis shows that PwNHX gene includes a coding region about 1350bp with 99% identity comparing with transposon Tn10 IS10-left transposase of Shigella flexneri. On the one hand, the NHX gene may lose its function because it was inserted a fragment in coding region. On the other hand, its product may play a important role in salt tolerance. Populus grow in saline soil. It speculates that it may have other salt tolerance mechanism in Populus. The transposon can be used as transposon tagging to clone other genes and it will help us to understand farther the salt tolerance mechanism.
Amino Acid Sequence ; DNA Transposable Elements ; genetics ; DNA, Complementary ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Plant Proteins ; chemistry ; genetics ; Populus ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Sequence Homology, Amino Acid ; Shigella flexneri ; genetics