The sfa1 gene encoded a bifunctional enzyme with the activities of both alcohol dehydrogenase and glutathione-dependent formaldehyde dehydrogenase in Saccharomyces cerevisiae.The gene disruption cassette produced by PCR using the same long oligonucleotides which comprise 19 or 22 nucleotides complementary to sequences in the templates(pUG6 and pUG66 marker plasmid)at 3' end and 45 nucleotides at 5' end that annealed to sites upstream or downstream of the genomic target sequence to be deleted.After two linear disruption cassettes with a Cre/loxP mediated marker were transformed into the cells of Saccharomyces cerevisiae YS-1,the positive transformants were checked by PCR to correct the integration of the cassette and concurrent deletion of the chromosomal target sequence.Once correctly integrated into the genome,the select marker can be efficiently rescued by transformating the plasmid pSH47 into YS-1 and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.The expression of the Cre recombinase finally resulted in the removal of the marker gene,leaving behind a single loxP site at the chromosomal locus.The diploid mutant YS-1-sfa1 was generated,which could enhance the output of ethanol with 8.0% by shaking culture in flask compared with the original strain YS-1.

Adolescent ; Child ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Killer Cells, Natural ; pathology ; Leukocyte Common Antigens ; analysis ; Lymphoma, T-Cell ; genetics ; immunology ; Male ; Microtubule-Associated Proteins ; genetics

OBJECTIVETo determine Verbascoside in Galeobdolon chinense.

METHODHPLC method was used, with the column packed with Kromasil C18(4.6 mm x 150 mm, 5 microns). The mobile phase was a mixture of acetonitrile and 1% acetic acid in a ratio of 13:87, and the ultraviolet wavelength was set at 350 nm.

RESULTThe average recovery was 97.0%, the RSD being 1.69%.

CONCLUSIONThis method is sensitive and reliable. It can be used to determine being Verbascoside in Galeobdolon chinense.

Chromatography, High Pressure Liquid ; methods ; Glucosides ; analysis ; Lamiaceae ; chemistry ; Phenols ; analysis ; Plants, Medicinal ; chemistry

Adolescent ; Adolescent Development ; Child ; Child Development ; China ; epidemiology ; Female ; Humans ; Male ; Obesity ; epidemiology ; Prevalence

The purpose of this investigation is to improve ethanol production and decrease acetate formation in Saccharomyces cerevisiae strain YS2-?adh2.The strain YS2-?adh2 with deleted alcohol dehydrogenase Ⅱ(adh2) gene was isolated in our lab with higher ethanol production than that of the strain YS2.The ace-taldehyde dehydrogenase Ⅵ(ald6) gene encoded a cytosolic acetaldehyde dehydrogenase,a key enzyme of the pyruvate dehydrogenase(PDH) bypass,transfers acetaldehyde to acetate.To disrupt ald6 gene of the strain YS2-?adh2,ald6 gene targeting cassettes were synthesized by long flanking homology PCR(LFH-PCR) and then were transformed into YS2-?adh2 mutants by LiAc/SS Carrier DNA/PEG method.Positive transformants were selected with G418 and further confirmed by PCR.Once correctly integrated into the genome,the selective marker was rescued by transforming the plasmid pSH65 into the positive transformants and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.We named the ald6 gene knocked-out strain as YS2-?adh2-?ald6 which has a 12.5% higher ethanol production and a 18% lower acetate formation compared to the strain YS2.

OBJECTIVETo characterize the meningococcal strains isolated from cases and close contacts with meningococcal disease associated with an outbreak in a jail in May 2010 by investigating the national distribution of hyperinvasive ST-4821 serogroup C clone associated with this outbreak.

METHODSThe cases were described based on the clinical symptoms and laboratory results. Pharyngeal swabs were cultured for N. meningitidis from men in the jail. Meningococcal isolates were identified by serogrouping, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST), respectively. Four hundred and sixteen serogroup C N. meningitidis strains were collected from 27 provinces between 2003 and 2010 for a nationwide survey and analyzed by PFGE and MLST.

RESULTSThree persons in a jail system were infected with invasive N. meningitidis serogroup C. All isolates tested had matching PFGE patterns and belonged to the multilocus sequence type (ST) 4821 clonal complex. All 47 N. meningitidis strains were identified from the pharyngeal swabs of 166 peoples in the jail, and 26 of them belonged to ST-4821 serogroup C clone, and 90.14% (375/416) serogroup C strains identified in the nationwide survey belonged to the ST-4821 complex. The ST-4821 serogroup C clone was spread nationwide, distributed in 24 provinces, especially in eastern provinces between 2003 and 2010.

CONCLUSIONEndemic transmission and carriage rate of ST-4821 serogroup C clone are high in this jail system. The ST-4821 serogroup C clone is spreading in China and nationwide distributed despite the existence of some effective vaccines.

Carrier State ; China ; epidemiology ; Disease Outbreaks ; Electrophoresis, Gel, Pulsed-Field ; Humans ; Meningitis, Meningococcal ; epidemiology ; microbiology ; Neisseria meningitidis ; genetics ; isolation & purification ; Pharynx ; microbiology ; Prisons

Objective To estimate the outpatient rate of influenza-related influenza-like illness (ILI) for children younger than 5 years in Suzhou municipal districts. Methods From October 2011 to March 2017, we conducted a prospective surveillance program on ILI for children under 5 years in outpatient settings of Soochow University Affiliated Children’s Hospital (SCH). The throat swabs were collected and tested for influenza viruses by RT-PCR. Based on the healthcare utilization surveys and population data, the number of visits and the outpatient rate of influenza-related ILI for children younger than 5 years in Suzhou municipal districts were estimated. Results During 2011-2017, in total, there were 45 930 estimated influenza-related ILI cases younger than 5 years in Suzhou municipal districts, which consisted of 7 490 influenza A/H1N1 cases, 17 843 influenza A/H3N2 cases and 20 597 influenza B cases. The estimated outpatient rate of influenza-related ILI was 6.4% in 2011-2017, which was highest in 2011-2012, 20.5%, and the lowest in 2012-2013, 2.4%. Conclusion The number of visits and the outpatient rate of influenza-related ILI in children younger than 5 years was high in Suzhou municipal districts.

Objective To understand the distribution and epidemic characteristics of common pathogens of pneumonia among hospitalized children in Suzhou． Methods Nasopharyngeal secretions were collected from hospitalized children with clinical pneumonia admitted to the respiratory department of Children＇s Hospital Affiliated to Suzhou University from April 2011 to March 2018 to detect common viral and bacterial pathogens of children＇s pneumonia． Ｒesults The total positive rate of pathogens was 75. 6% in the 4 765 clinical pneumonia cases. The positive rate of bacterial pathogens was 57. 4%. Streptococcus pneumoniae ( SP) was the highest，followed by Haemophilus influenzae ( H. i) ; The positive rate of viral pathogens was 44. 1%. Ｒespiratory syncytial virus ( ＲSV) was the highest，followed by Bocavirus ( BoV) . The mixed infection rate of bacteria and virus was 25. 9%，and the most common types were ＲSV and SP，BoV and Streptococcus viride ( SV) ． Conclusions SP，H．i，ＲSV and BoV are the main pathogens of clinical pneumonia in children． There are statistical differences in different age groups and seasons of hospitalized children＇s pneumonia in Suzhou． The mixed infection rate of bacteria and virus is high．

Objective: To explore the clinical characteristics and prognostic factors of the patients with non-Hodgkin's Lymphoma (NHL) complicated with HBV infection, so as to provide a basis for clinical accurate diagnosis and prognosis evaluation. Methods: The data of 313 newly diagnosed NHL patients from August 2012 to July 2016 were collected. The HBV serological markers were detected by ELISA, and HBV DNA was quantified by full automatic microparticle chemiluminescence immunoassay (≥1×10(5) copies/ml as high copy group, 1×10(3)-<1×10(5) copies/ml as low copy group). The relationship between HBV infection and prognosis was analyzed combined with the clinical features of the patients, and the HBV detection rate was compared with that of the common population (from the national HBV sero epidemiological data). Results: ①The positive rate of HBsAg in NHL patients was 12.5% (39/313), which was higher than 7.2% in the general population (χ(2)=14.596, P<0.001). HBV infection in the past (HBsAg negative but HBcAb positive) in 114 cases (36.4%), the incidence was slightly higher than that in the general population (34.1%). ②Compared HBsAg positive group with the negative group, the proportion of B cell type (87.2% vs 70.3%, P=0.027), Ann Arbor stage Ⅲ-Ⅳ(69.2% vs 34.6%, P<0.001), IPI score 3-5 (74.4% vs 50%, P=0.004), LDH level (79.5% vs 47.8%, P<0.001) and liver involvement (45.5% vs 31.7%, P=0.006) were all higher. The difference was statistically significant. ③Compared the HBV infected group (114 cases) with the non-infected group (160 cases), the difference had statistical significance in the proportion of Ann Arbor stage Ⅲ-Ⅳ (P=0.023) and IPI score 3-5 scores P=0.035). ④Compared HBV DNA positive group (30 cases) with negative group (71 cases), Ann Arbor stage Ⅲ-Ⅳ (P=0.011), IPI score 3-5 score (P=0.030), LDH level (P=0.025) and liver involvement (P<0.001) in the proportion of patients had statistical significance. The positive patients were divided into HBV DNA high and low copy groups with 1×10(5) copies of /ml as the boundary. The results showed that there was no statistical difference between the two groups (P>0.05). Conclusions: The HBV infection rate of NHL patients is significantly higher than that of the general population, and HBV infection is more closely related to B cell type NHL. Patients with HBV infection and HBV DNA positive had late Ann Arbor stage, high IPI score, high LDH level and liver involvement, and the prognosis is poor.
Hepatitis B ; Hepatitis B Antibodies ; Hepatitis B Surface Antigens ; Humans ; Lymphoma, Non-Hodgkin ; Prognosis