[Analysis on the whole genome of the influenza H1N1 virus of the mild and severe cases in Beijing in 2009].

OBJECTIVE: To explore the characteristics of the whole genome of the influenza H1N1 virus of the mild and severe cases in Beijing. METHODS: A total of 21 samples of throat swabs were collected from surveillance-designated hospitals between June and December in 2009, including 10 severe cases (4 death cases) and 11 mild cases. RNA of the virus were extracted,and the amplified primers of the whole genome were designed.Reverse transcription and PCR were performed to the RNA and then the PCR product was sequenced by software to analyze the evolution of the viral genes and the variation of the amino acids. RESULTS: Compared with the reference vaccine strain A/California/07/2009 (H1N1), the genetic nucleotide homology in the eight segments of the pandemic H1N1 virus in Beijing in 2009 was higher than 99%, without significant variation. Among them,the genetic distance of hemagglutinin (HA), neuraminidase (NA) and nucleoprotein (NP) was comparatively far, separately 0.0050, 0.0040 and 0.0040.The gene of HA, P83S, the gene of NA, N248D, the gene of polymerase (PA), P224S and the gene of NP, V100I and L122Q were found to mutate in all the samples. Genes of HA, NA, NP, PA, PB 2 and nonstructural protein (NS1) in severe cases showed obviously clustered evolution. The mutation of gene S128P and S203T of HA, gene R269R and D547E of PA, gene T588I of PB 2 and gene I123V of NS mainly happened in severe cases, separately counting 6, 9, 6, 7, 9 and 6 cases. The relevance between the mutation happened in S203T of HA, R269K and D547E of PA and the severeness of the cases showed statistical significance (P < 0.05). The mutations of HA gene were mainly on the Ca and Cb antigene domains. No drug resistant mutation was found on NA gene but happened on matrix protein 2 (M2 gene). None of the mutations were found on the virulence related genes. CONCLUSION: A high homology was found between the pandemic H1N1 virus in Beijing in 2009 and the reference vaccine strain A/California/07/2009(H1N1). Mutational sites related with the severe and fatal cases were found, but not the virulence related mutation.

[Application of EARS in early-warning of influenza pandemic in Beijing].

OBJECTIVE: To illustrate the efficiency of cumulative sum (CUSUM) in pre-warning of the influenza peak in Beijing. METHODS: CUSUM was used to analyze the data of influenza like illness (ILI), and the results of the influenza laboratory surveillance was regarded as the gold standard to judge the approaching of the influenza peak. RESULTS: The surveillance was launched in 421 hospitals in Beijing during the 2009 to 2010 influenza season, while the influenza laboratory surveillance was launched by 7 collaborative laboratories. From Jun. 2009 to Apr. 2010, the average ILI percentage in the 421 hospitals was 2.56%. In the study, 19 262 pharyngeal swab samples were collected from the ILI cases in 11 hospitals and 5 045 of them were tested positive for the influenza virus, with the novel swine-origin influenza A H1N1 virus dominating. After analyzing of the ILI surveillance data with CUSUM, it was found that the ILI surveillance in Beijing could make a satisfactory early warning for the approaching of the influenza peak referring to the gold standard based on the influenza laboratory results. CONCLUSION: It could give the prediction and early warning for the influenza peak efficiently and precisely, by using CUSUM to analyze the influenza surveillance data of Beijing.

[Study on types of M protein gene in group A streptococcus isolated from children in Beijing, 2011].

OBJECTIVE: To explore the distribution characteristics of the types of M protein gene (emm) in group A streptococcus (GAS) isolated from children in Beijing in the year 2011. METHODS: During May to July in 2011, a total of 3315 patients who were diagnosed scarlet fever or pharyngeal infection by doctors in pediatric outpatient and emergency units of 36 hospitals, were selected as subjects. Their throat swab samples were collected and isolated the strains of GAS. Gene emm was then amplified and sequenced by PCR method, and the differences in types of gene emm between different populations and diseases were compared. RESULTS: A total of 633 strains of GAS were isolated from the 3315 throat swab samples, 610 strains out of which were gene emm positive and were recruited in the study. Out of the 610 recruited strains, 448 (73.4%) were isolated from scarlet fever patients, the other 162 (26.6%) were isolated from pharyngeal infection patients; 397 (65.1%) were from urban, the other 213 (34.9%) were from suburb; 240 (39.4%) were from patients aging between 1 - 5 years old, the other 369(60.6%) were from patients aging 6 - 18 years old. A total of 8 types of gene emm (scarlet fever: 6 types, pharyngeal infection: 4 types) and 21 subtypes of gene emn (scarlet fever: 16 subtypes, pharyngeal infection: 10 subtypes) were identified. Three new subtypes were found in the study, naming emm1.63, emm12.62 and st5144.20. Among them, emm1.63 was found both in scarlet fever and pharyngeal infection patients, while emm12.62 and st5144. 20 were only found in pharyngeal infection patients. Among all the types of gene-emm, emm12 accounted for the highest percentage as 80.5% (491/610) and then followed by emm1 (18.0% (110/610)). Among all the subtypes, the dominant subtype was emm12.00, accounting for 69.0% (421/610), following by emm1.00 (16.9% (103/610)) and emm12.19 (6.1% (37/610)). All the above types and subtypes of gene emm were the most prevalent strains in scarlet fever patients and pharyngeal infection patients. Significant differences in the distribution of prevalent strains were observed among various aging patients and regions. The constituent ratios of emm1, emm1.00 and emm12.19 were higher in patients from suburb (emm1: 22.1% (47/213), emm1.00: 19.2% (40/213), emm12.19: 8.0% (17/213)) than those in urban areas (emm1: 15.9% (63/397), emm1.00: 15.6% (62/397), emm12.19: 5.0% (20/397)). The difference showed statistical significance (P < 0.05). The constituent ratio of emm1.00 was higher among patients aging 6-18 years old (19.2% (71/369)) than those aging 1 - 5 years old (13.3% (32/240)). The difference also showed statistical significance (χ(2) = 8.45, P < 0.05). CONCLUSION: Among the types of gene emm in GAS isolated from children in Beijing in year 2011, the most prevalent two were emm12 and emm1, and the most prevalent emm subtypes were emm12.00, emm1.00 and emm12.19. A significant difference in their distribution between various aging patients and isolated places can be obviously found.

[Bioremediation of petroleum hydrocarbon-contaminated soils by cold-adapted microorganisms: research advance].

Cold-adapted microorganisms such as psychrotrophs and psychrophiles widely exist in the soils of sub-Arctic, Arctic, Antarctic, alpine, and high mountains, being the important microbial resources for the biodegradation of petroleum hydrocarbons at low temperature. Using the unique advantage of cold-adapted microorganisms to the bioremediation of petroleum hydrocarbon-contaminated soils in low temperature region has become a research hotspot. This paper summarized the category and cold-adaptation mechanisms of the microorganisms able to degrade petroleum hydrocarbon at low temperature, biodegradation characteristics and mechanisms of different petroleum fractions under the action of cold-adapted microorganisms, bio-stimulation techniques for improving biodegradation efficiency, e. g., inoculating petroleum-degrading microorganisms and adding nutrients or bio-surfactants, and the present status of applying molecular biotechnology in this research field, aimed to provide references to the development of bioremediation techniques for petroleum hydrocarbon-contaminated soils.

[A survey on serological epidemiology of influenza A (H1N1) 2009 in Beijing].

OBJECTIVE: To investigate the immunological level against influenza A (H1N1) 2009 in Beijing and provide evidence to evaluate the developing trend of the disease. METHODS: Between Nov. 27, 2009 and Dec. 23, 2009, subjects were randomly selected from patients in hospitals (infectious and respiratory diseases related departments were excluded), volunteers in blood donation center and healthy subjects attending the physical examination center. Questionnaire survey was conducted and serum samples were collected to detect the hemagglutination-inhibition (HI) antibody against influenza A (H1N1) 2009 virus. RESULTS: 856 subjects participated in this survey, and 127 showed positive HI antibody to this pandemic virus. The proportions of sero-positivity among 0 - 5, 6 - 17, 18 - 55, ≥ 56 year olds were 14.5%, 19.4%, 17.4% and 8.0% respectively (P = 0.009). There was no significant difference in the sero-positivity between males and females (P = 0.693). The age-adjusted positive rate was 15.8% in the population of Beijing. By multivariate logistic regression analysis, factors as age, acute respiratory symptoms and the rate of pandemic (H1N1) 2009 vaccination were significantly associated with sero-positivity of HI antibody to the influenza A (H1N1) 2009 virus. CONCLUSION: Above 15% of the population in Beijing showed protective antibody against influenza A (H1N1) 2009 virus, indicating the development of immunological barrier to this disease had been formed, to some extent.

[Etiological characteristics of influenza A (H1N1) 2009 virus in Beijing].

OBJECTIVE: To analyze the results of detection on influenza A (H1N1) 2009 virus in Beijing from May 2009 to December 2009 and to understand the epidemiologic characteristics during the pandemic period. METHODS: The study was conducted from the May 1 to December 27, 2009. A total of 101 852 throat swab samples were detected with the real-time RT-PCR assay by the Beijing Network Laboratory. Data was statistically analyzed. RESULTS: 9843 samples showed influenza A (H1N1) 2009 positive, with an overall positive rate as 9.66%. In terms of the positive rates, they were 2.85% from May to June, 3.32% from July to August and 8.35% from September to October. The peak month fell in November (29.67%) and December (24.33%). The positive rates among the following subpopulations were: 8.40% among the suspected cases, 4.75% among close contact cases, 11.46% among the influenza-like illness cases and 7.33% among the cluster cases with fever. Positive cases mainly fell in age groups 5 - 14 and 15 - 24. The ratio of male to female was 1.5:1. CONCLUSION: During the pandemic period of influenza A (H1N1) 2009, positive cases gradually increased during May to November but slowly decreasing in December.

[The significance of different sample types in study of pandemic A (H1N1) influenza diagnosis].

OBJECTIVE: To explore the value of different types of samples, including throat swabs, stools, bloods in pandemic A (H1N1) influenza diagnosis and virus shedding patterns. METHODS: From May to June in 2009, 135 samples were collected from 23 confirmed cases of pandemic influenza A (H1N1) infection, including 99 throat swabs, 14 stools, 11 bloods, 1 respiratory tract washing from 13 confirmed cases and 10 blood samples from other confirmed cases. The virus was detected by real-time RT-PCR, the antibody was detected by haemagglutination inhibition assay. RESULTS: For 99 throat swabs of 13 patients, the median time of the first positive real-time RT-PCR was 1 day (ranged from 0 to 7 days) after the onset of the symptoms of illness; the median length of time duration of positive real-time RT-PCR results from throat swabs was 3 days (ranged from 1 to 15 days). Four cases intermittently released virus. One respiratory tract washing sample was positive. In 14 stools, 8 stools were real-time RT-PCR positive, the positive rate was 57.14%. The median time of the positive real-time RT-PCR was 3 days (ranged from 1 to 4 days) after the onset of the symptoms of illness. In 21 blood samples collected at 2 to 9 days of onset, 1 blood sample was real-time RT-PCR positive, the positive rate was 4.76%. All these 21 blood samples were antibody negative. CONCLUSION: Throat swabs and stools samples can be used as A (H1N1) influenza early diagnosis. The length of time duration of positive real-time RT-PCR in throat swabs was longer than stool samples and intermittently releasing of virus were found in throat swabs. Influenza A H1N1 cases showed the presence of small amount of viremia and antibody was negative in early blood samples (< 9 days).

Ecotoxicity assessment of aged petroleum sludge using a suite of effects-based end points in earthworm Eisenia fetida.

Laboratory investigations were conducted to test the toxicity of aged petroleum sludge collected from Shengli Oil Field, the second largest oilfield in China, to earthworm Eisenia fetida. Various end points were measured in the earthworms, including mortality, growth, cocoon output, juvenile production, and avoidance behavioral response, to determine their comparative sensitivity for assessing harmful effects of petroleum-hydrocarbon-contaminated soil. The results showed that all these assays responded in a concentration-dependent manner, and two chronic end points, juvenile production and cocoon output, as well as avoidance behavioral response appeared to be sensitive end points for detecting toxicity of petroleum-hydrocarbon-contaminated soil. Comparatively, juvenile production exhibited similar sensitivity to avoidance behavior response, both of which were more sensitive than cocoon output, while mortality and adult growth were proposed as the least sensitive parameters. It was suggested that large amounts of petroleum sludge deposited in Shengli Oil Field may pose a potential threat to the local ecosystem, and the utility of multiple effects-based end points in earthworm E. fetida is useful to facilitate ecological risk assessments in hydrocarbon-contaminated sites.