Next generation sequencing technologies,providing high speed and throughput that contribute greatly to DNA sequencing,have many applications in clinical virology,including identification of novel or unsuspected viruses,evaluation of human viromes,investigation of viral genetic diversity and evolution,monitoring of antiviral drug resistance,and analysis of virus-host interaction.Meanwhile,due to the complexity in bioinformatics data processing and lack of quality management standards,further efforts and explorations need to be put in the application and quality management of NGS in clinical laboratory.

Objective To obtain the molecular epidemiology of human Parechovirus (HPeV)infections m children with central nervous system (CNS)-related disease and sepsis,as well as understand the pathogenic properties of HPeV infections by detecting HPeV in cerebrospinal fluid (CSF) and blood samples.Methods From January to December in the year of 2009,a total of 359enterovirus-negative specimens including 210 CSF and 149 blood samples were collected from 328children ＜14 years of age who were hospitalized for CNS-related disease and sepsis at Children's Hospital,Fudan University,Shanghai,China.HPeV was detected by nested reverse transcription polymerase chain reaction (RT-PCR),and then directly genotyped by sequencing nested RT-PCR product of VP3/VP1 region.Ninty-nine blood samples from healthy children were collected as controls during the same period.Results Twenty-seven children (8.2％) were HPeV positive in 328 children.HPeV infections were found in all age groups of children and the highest frequency was seen in children ＜3 months old (18.2％,12/66).HPeV was detected in several months,with the peak in December (18.8％,9/48).Of all the positive samples,20 were genotyped successfully and identified to be HPeV1.No HPeV infections were found in blood of healthy controls.ConclusionsHPeV is the pathogen of CNS infections and sepsis in children.HPeV screening should be enrolled in the routine virus testing in specimens obtained from children.HPeV1 is the prevalent type in children in the year of 2009 in Shanghai.

Objective To study effect of Xuebijing injection on coagulation function and prevention of deep venous thrombosis in elderly fracture patients with mechanical ventilation.Methods 60 patients were randomly divided into control group(30 cases) and Xuebijing group(30 cases).Xuebijing group received Xuebijing injection, besides physical therapy for prevention of deep venous thrombosis received by control group.Coagulation parameters and Lac at different time points,thrombosis incidence,hemorrhage incidence APACHE II score and 28 -day mortality were compared between the two groups.Results In Xuebijing group,PT,APTT,DD on d3 and Fg 48 on d5 had statistically significant differences compared with before treatment.PT,APTT on d3,d5 and DD on d5 were statistical-ly different compared with the control group(P <0.05).PLT,Lac,hemorrhage incidence and 28 -day mortality had no significant differences between the two groups.Thrombosis incidence in Xuebijing group was significantly lower than the control group(20.00% vs 3.33%) (χ2 =4.043,P =0.044,P <0.05).Both of APACHE II score were lower than before treatment,but the Xuebijing group was significantly decreased(t =5.48,P =0.000,P <0.05). Conclusion Xuebijing injection could improve coagulation function,and decrease thrombosis incidence in elderly fracture patients with mechanical ventilation.

Objective To observe the genetic structure of cultivated Scrophularia ningpoensis in Zhejiang Province.Methods The genetic structures of six typical S.ningpoensis populations were analyzed by fluorescence AFLP marker.Results Bands(12 552) were generated by seven pairs of AFLP primer combinations,of which 8 808 were polymorphic,and the polymorphic rate was 70.17%.The variety ranges of PPB among different populations were 41.67%—55.56%,and 47.30% in average.I was between 0.190 8—0.238 3,and 0.221 8 in average.Ne was between 1.201 4—1.280 6,and 1.236 9 in average.Gst was 0.127 1,Nm was 3.432 4.UPGMA Cluster analysis showed that the six populations can be divided into two clusters,as that of Tiantai,Jinyun,and Jingning were one sub-cluster,and Dongyang,Pan′an,and Xianju were another one sub-cluster.Conclusion There is a relative high genetic diversity level in cultured S.ningpoensis of Zhejiang Province.Genetic differentiation exists among populations,but it exists in population mostly.There is a relative high genetic intercommunion among populations.The genetic distance is not related to the geographic environment.

Objective To investigate the dynamic characteristic of molecular epidemiology of group A Rotavirus (RV) by analyzing viral genotypes,disease seasonality,and the patients' age distribution,so that to provide theoretical basis for preyention and control of RV diarrhea in children.MethodsA total of 380 RV antigen positive samples were selected from 5176 stool specimens collected from ＜5 year-old patients with acute diarrhea who were admitted to Children's Hospital of Fudan University during January 2006 to December 2008. Multiplex nested reverse transcription polymerase chain reaction (RT-PCR) was used to analyze the RV genotypes.ResultsDuring 2006-2008,the incidence of RV related diarrhea peaked from October to December and about 96.8％ of all RV episodes occurred in patients younger than 3 years old,The predominant genotype was G3 which accounted for 58.4％ (222/380),G9 was an emerging genotype with the prevalence rate as high as 10.8％ (41/380).G1 and G2 types were rarely found during the three years.Infections with both G3 and G9 were the major mixed genotype G infection. Genotype P [8] was predominant with the prevalence rates of 64.6％ (53/82) and 46.8％ (58/124) in 2006 and 2008,respectively,whereasgenotype P[4] was predominant in 2007 (38,5％,67/174).P[6] and P[9] were found as minor types.The major mixed genotype P infection were genotype P[4] and P[8]. The proportion of undetermined genotype G and genotype P strains tended to increasing during 2006-2008.Genotype P [8]G3 was the major RV strain (20.5％) in Shanghai during 2006-2008 and the other prevalent genotypes included P[4]G3 and P[m]G3.Conclusion The infection of group A RV in Shanghai presents some new molecular epidemiology characteristics during 2006-2008,such as switch of predominant genotypes and diversification of prevalent genotypes.

Objective To compare the molecular epidemic characteristics of human astrovirus (HAstV) between outpatient and hospitalized children with acute diarrhea,and to investigate the relationship between HAstY infection and diarrhea in children.Methods A total of 298 cases were randomly collected from hospitalized children from January 2008 to December 2010 in Children's Hospital of Fudan University,and 360 specimens were collected from outpatients with acute diarrhea from August 2010 to July 2011.Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect rotavirus (RV),human calicivirus (HuCV),HAstV and human adenovirus (HAdV).H AstV genotype was determined by gene sequencing and phylogenetic analysis.Results Epidemiology of HAstV in hospitalized children was as follows:among the included 298 samples,HAstV was detected in 27.2％ (81/298) of the patients,compared with 33.9％ (42/124),33.8％ (25/74) and 14.0％ (14/100),respectively from 2008 to 2010.HAstV diarrhea occurred throughout the year and peaked in January,March,and April.95.1％ (77/81) of the infected children were 0-35 months old.All the episodes of HAstV were mixed with other diarrhea virus infection.Molecular epidemiology of HAstV in outpatient children with diarrhea was as follows:the overall incidence of HAstV was 1.9 ％ among the 360 cases (7/360).The seasonal distribution of HAstV's gastroenteritis showed a peak in November.All the outpatient children were 0-35 months old.Three cases were single infection with HAstV and the others were coinfection with RV (3 cases) or HAdV (1 case).All of the detected HAstV,either in inpatients or outpatients,belonged to HAstV-1.Conclusions The detection rate of HAstV in hospitalized children is significantly higher than that in outpatients.Most HAstV infections in hospitalized children are ascribed to nosocomial infections.Most episodes of HAstV infection were accompanied with other diarrhea viruses infection.HAstY single infection is seen in outpatient children while the detection rate is very low,implying that HAstV co-infection with other viruses plays a main role in diarrhea in most instances.

Objective: To investigate the protective effect of rhubarb (大黄) combined with scopolamine on gastric and gut mucosal barrier in patients with severe trauma. Methods: Forty patients with severe trauma, injury severity score (ISS)≥25, were randomly divided into two groups (each n=20), namely common therapy group (CM group) and rhubarb (infused into stomach) combined with scopolamine group (RS group). In RS group, rhubarb combined with scopolamine were given just after the first time measuring gastrointramucosal pH (pHi), the other treatment was the same as CM group.The mean arterial pressure (MAP) and heart rate (HR) were recorded, and pHi, lactic acid, interleukin-6 (IL-6) and tumor necrosis factor-? (TNF-?) were measured on admission and at 24, 48 and 72 hours after admission. Results: The MAP, HR and pHi were uptrend in both groups. There were significant increase in MAP and pHi of RS group at 72 hours compared with those of CM group (both P

Objective To investigate the prevalence and attitude of smoking and its influencing factors among residents in Hongkou District of Shanghai, so as to provide evidence for tobacco control. Methods Nine hundred and sixty residents in Hongkou District of Shanghai aged 15 to 69 years were selected by multi-level random sampling, among whom 958 completed the investigations on smoking status. Results The smoking rate and standardized smoking rate were 20. 04% and 18. 73%, respectively. The smoking rate and standardized smoking rate of males were 41.05% and 30.07%, and those of females were 0. 80% and 1. 70% , respectively. The smoking rate of males was higher than that of females (RR = 0.02, 95% CI: 0.01 -0.03). The smoking rates of age groups of 25 years old and over were higher than those of 15 to 24 years old, and the smoking rate of subjects with or above junior college education was lower than that of subjects with or below middle school education (RR=0.45, 95% CI: 0.25 -0.79). Subjects who drank alcohol smoked more than those did not (RR = 2. 68, 95% CI: 1. 76 - 4.09). Lower annual per capita income and higher knowledge on smoking hazard led to more tendance to object to smoking and agree with anti-smoking. Conclusion The prevalence of smoking of residents aged 15 to 69 years in Hongkou District of Shanghai was relatively lower. Age, gender, education status and alcohol drinking were influencing factors of smoking. Education on tobacco control among subjects aged more than 35 years should be strengthened, and more attention should be paid to those with lower education. Besides, education on alcohol control should be managed together with tobacco control.

Objective The immune hypofunction of the red blood cell on maintenance hemodialysis patients is one main reason of tumor,and oxidative stress is proved to be involved in decrease of existence quality and the occurrence of various complications of patients with hemodialysis.To investigate the status of erythrocyte immune and high oxidative stress of the hemodialysis patients,as well as the effect of Huangqifuzitang on immune function of red blood cell and oxidative stress in patients with hemodialysis.Methods Twenty healthy people were chosen as control group,and 20 patients were selected as the treatment group.CD58,CD59,superoxide dismutase(SOD),malonaldehyde(MDA) were measured before hemodialysis and after 12 days administration of Huangqifuzitang in the treatment group,and compared with those of the control group.Results There was significant difference between before and after taking Huangqifuzitang in treatment group and control group in terms of CD58 ((38.02 ±8.98) vs.(47.39 ±7.78) vs.(59.10±4.59),F=4.506,P=0.000),CD59 ((62.69 ± 20.84) vs.(80.95 ± 20.42) vs.(193.86 ± 19.87) ; F =239.347,P =0.000),SOD ((68.09 ± 11.86) vs.(78.73±10.58) vs.(111.09±16.61) kU/L;F=21.318,P=0.000),MDA((5.98±2.06) vs.(4.54 ±0.62) vs.(3.03 ± 1.10) μmoL/L;F =55.359,P =0.000) levels.Before taking Huangqifuzitang,the concentration of CD58,CD59 and SOD in treatment group was significant decreased compared with control group,but MDA significant increased,and there was significant difference (P ＜ 0.01).After 12 days administration of Huangqifuzitang in the treatment group,the concentration of CD58,CD59 and SOD was significant increased compared with before taking Huangqifuzitang,but MDA significant significant decreased,and there was significant difference (P ＜ 0.01 or P ＜ 0.05) Conclusion The red blood cell surface receptors CD58,CD59 decreased and oxidative stress index SOD decreased significantly,MDA increased significantly in hemodialysis patients.Huangqifuzitang was proved to be with the ability of enhancing the immune of red bloodCD58,CD59 and decreasing the oxidative stress level in patients with hemodialysis.

Objective To investigate the MRI features of supratentorial solid hemangioblastoma and to improve its diagnostic accuracy.Methods MRI image manifestations of 7 cases with supratentorial solid hemangioblastoma confirmed by operation and pathology were analyzed retrospectively.Results 7 cases of supratentorial solid hemangioblastoma were solitary.All tumors appeared as round or quasi-round masses with clear boundary, and showed equal or long T1 signal and slight long or long T2 signal,the obvious enhancement was seen in all tumors.There was mild to moderate edema around the tumors.The blood vessel was found within or around the tumors.Conclusion MRI manifestations of supratentorial solid hemangioblastoma have certain characteristics.