Objective To understand the epidemiology of respiratory syncytial virus(RSV),influenza virus type A and type B(IV-A and IV-B),parainfluenza virus type 1,2,3(PIV-1,2,3)and adenovirus(ADV) among children with acute respiratory tract infection in Shanghai.Methods A retrospective epidemiological investigation was conducted to analyze the prevalence rate,seasonality and susceptible pediatric population of seven common respiratory viruses among hospitalized pediatric patients(<15 years old) with acute respiratory tract infection in Children's Hospital Affiliate to Fudan University Shanghai during 2003 to 2006.Nasopharyngeal aspirates were collected for detecting viral antigens of RSV,IV-A,IV-B,PIV-1,PIV-2,PIV-3 and ADV by direct immunofluorescence assay.Results Total 11214 children were collected during the consecutive four years,among whom 98.7% were presented with acute lower respiratory tract infection.The overall positive detection rate of these seven respiratory viruses was 24.2%.RSV accounted for 17.7%,followed by PIV-3(2.8%),ADV(2.2%),IV-A(0.7%),PIV-I(0.5%),PIV-2(0.3%),IV-B(0.1%)and mixed(0.2%).RSV season often occurred in winter and spring,however,an early season began in autumn every two year.PIV-3,ADV and IV were usually prevalent sporadically and the seasonal patterns were not typical.The median ages of infected pediatric patients were 4 months for RSV,8 months for PIV-3,9.5 months for PIV-1,10.5 months for PIV-2,12 months for ADV,13 months for IV,respectively,which were significantly different by statistical analysis(X2=154.319,P<0.01).The infants and younger children were more susceptible for developing RSV and PIV-3 related diseases.RSV infection significantly decreased along with the age increased.Conclusions RSV is the most commonly identified virus,while infants and neonates are the most susceptible populations for RSV infection.Typically,RSV is prevalent during spring and winter but begins to be active in fall every other year. PIV-3 is found to be the second common viral pathogen and young children are susceptible population.ADV ranks the third pathogen and children with ADV infection tend to be elder. Nevertheless,IV is rare among pediatric population in recent years in Shanghai.In our study,the outbreaks of ADV and IV infections among children are not observed.

Objective To study the clinical and the epidemiological features of hospitalized children with influenza virus infection. Methods Two hundred and fifty-three inpatients with laboratory-confirmed influenza virus infection from 1999 to 2008 were reviewed for analyzing the clinical and epidemiological characteristics. Type A and B influenza viruses in the nasopharyngeal aspirates were detected by immunofluorescence assay. Mann-Whitney U test were performed for comparing the median age and the length of hospitalization. Chi-square test was performed for comparing the proportion of patients with fever and cough. Results Among 253 hospitalized children aged between 5 days and 127 months, 182 (71.9%) were boys and 71 (28. 1%) were girls. The median age was 18 months. Fifty-three cases were infants younger than 6 months. 95 cases were children aged between 6 months and 2 years, 85 cases were aged between 2 years and 5 years and 20 cases were older than 5 years. The diagnosis of influenza-related admission included pneumonia (190 cases), bronchitis (49 cases) and upper respiratory tract infection (14 cases). Eleven cases developed febrile convulsion, 6 cases had acute exacerbation of asthma and 3 cases had concomitant viralencephalitis. Twenty-nine cases had basic diseases. Cough and fever were the most common symptoms. Two hundred and thirty-eight cases presenting cough and 209 case presenting fever. Sixty-seven percent (140/209) had high fever with body temperature higher than 39 ℃. The average duration of fever was (5. 0 ±2. 9) days. Fever and cough were both more common in children older than6 months (X2 = 22. 895,P<0. 01; X2 = 16. 992,P<0. 01, respectively). Febrile convulsion occurred in children older than 2 years. Fifteen point five (39/251) developed leukocytopenia. Conclusions Children younger than 5 years old are at high risk of influenza-related hospitalization. We should emphasize influenza vaccination in previously healthy children aged between 6 months and 5 years and children with underlying diseases.

To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.
Animals ; Biological Transport ; Cimetidine ; pharmacology ; DNA, Complementary ; Dogs ; Drug Interactions ; Humans ; Madin Darby Canine Kidney Cells ; Metformin ; pharmacology ; Organic Cation Transport Proteins ; genetics ; metabolism ; Transfection

Objective To understand the erythromycin resistance rate and the erythromycin resistant gene spectrum in Streptococcus pyogenes strains isolated in Shanghai.Methods The outpatient children who were diagnosed with scarlatinal in the Children's Hospital of Fudan University from November 2004 to June 2006 were enrolled and 100 strains of Streptococcus pyogenes were isolated by pharyngeal swab culture.The distributions ofermA,ermB,mefA genes were determined by polymerase chain reaction(PCR)amplification.The relationship between ermA,ermB,mefA genes and erythromycin resistance were also analyzed.Results The erythromycin and clindamycin resistance rates of Streptococcus pyogenes in Shanghai were 98%and 95%,respectively;the concordance rate of these two drugs was 97%.Among 100 strains of Streptococcus pyogenes,94 strains(94%)contained ermB gene,with 100%of erythromycinresistance rate.Sixteen(16%)contained mefA gene,with 100% of erythromycin resistance rate.ermA was not detected inall the 100 strains.The ermB and mefA genes were not found in 5 strains,among which,2 were susceptible to erythromycin and 3wereresistant to erythromycin.Only 1%of isolates was mefA genesingle positive.Conclusions There is a high erythromycin resistance rate of Streptococcus pyogenes strains isolated inShanghai,and the cross resistance to clindamycin is high.TheermB gene is important erythromycin resistancedeterminants of Streptococcus pyogenes in Shanghai.

Objective To evaluate engraftment status of patients after allogeneic hematopoietic stem cell transplantation(Allo-HSCT) and prompt relapse of disease based on DNA polymorphism analysis technique.Methods Sixty-six cases were detected by DNA polymorphism analysis technique and 25 cases were monitored and analyzed dynamically during this period.Results After Allo-HSCT,48 patients obtained type of donors,but 13 patients did not; 5 patients showed mixed chimerism.Two cases of type of donors converted into mixed chimerism and 4 cases of mixed chimerism converted into type of donors after some time. The others' engraftment status did not change.Conclusion DNA polymorphism analysis technique can detect engraftment status of patients exactly, rapidly, which provides effective evidences of constitution for more clinical therapy projects.

Objective To explore the clinical characteristics and emergency treatment for type 1 diabetes(T1DM) and diabetes ketoacidosis(DKA) in children under 5 years old.Methods Twenty-one children under 5 years old with T1DM with 10 years were retrospecti-vely reviewed.The onset situation,clinical feature and treatment of DKA were analyzed.Results The cases of little children diabetes might not have typical symptoms.The positivity of islet antibody was lower.High morbidity of DKA was found in little children and DKA was often caused by infection.Conclusions Infection may be involved in the onset and progress of childhood T1DM.Emergency treatment for DKA may involve the injection of small dose insulin,correction of the disorder of water and electrolysis and regulation of acid-base.

Objective To elucidate the prevalence and clinical characteristics of human metapneumovirus(hMPV)in hospital- ized children with respiratory infection.Methods A total of 452 hospitalized children with lower respiratory tract infection were observed from Aug 2004 to Jan 2005.Respiratory tract aspirates were collected from all patients within 48 hours after admis sion.The specimens were routinely tested for respiratory syncytial virus,influenza virus A and B,parainfluenza virus 1 to 3 and adenovirus by direct fluorescent assay(DFA).The 245 specimens negative by DFA were tested for hMPV by RT-PCR. PCR products of hMPV M gene from some patients were randomly selected for sequencing analysis.Results hMPV was identi- fied in 59(24.1%)of the 245 specimens tested,hMPV infection alone accounted for 13.1% of the infections in the 452 chil- dren under study,The prevalence of hMPV was higher than other respiratory viruses in winter.The mean age of hMPV-infec- ted children(n=59)was 27.7 months.There was no significant difference between age groups in terms of the prevalence of hMPV(P＞0.05).There were no statistically significant difference in demographics and clinical symptoms between hMPV in- fection and other common respiratory virus infection.Genotyping for the hMPV M gene from 23 Shanghai patients showed two distinct hMPV genotypes.Sequence analysis of these hMPV M genes showed 82.8%-100% homology to the registered se- quence in GenBank.There was no significant difference in clinical characteristics between the 2 genotypes.Conclusions hMPV plays an important pathogenic role in lower respiratory tract infection of children,hMPV prevailed in the winter of 2004.Clini- cally,hMPV infection can not be discriminated from the infection of other respiratory viruses.Clinical manifestation is similar between the two hMPV genotypes.

Objective To observe the effects of Xifeng Tongnao capsules on the content of TNF-α and IL-1β in brain tissue of rats with focal cerebral ischemia-reperfusion injury. Methods A total of 120 adult SD rats were randomized into 6 groups: Xifeng Tongnao low-, middle- and high-dose groups, model control group, Buchang Naoxintong group and sham-operated group.Buchang Naoxintong group were treated with Buchang Naoxintong capsules at 0.864 g??kg-1.Xifeng Tongnao high-, middle-and low-dose groups were treated with 3.456, 1.728, and 0.864 g??kg-1 Xifeng Tongnao capsules, respectively;sham-operated group and model control group were treated with equal volume of purified water. Medications were administered intragastrically once daily for 7 days. The acute transient focal cerebral ischemia-reperfusion model was established by middle cerebral artery occlusion ( MCAO) 1 h after the final dose, and rats in the sham-operated group only received anesthesia and stripping without occlusion.All rats were sacrificed after reperfusion for 24 h, and expression levels of TNF-α and IL-1β in the brain tissue were detected by ELISA. Results TNF-αcontent in Xifeng Tongnao capsutes low-, middle-and high-dose groups were (35.34±8.95), (33.75±6.92), and (40.95±5.39) ng??L-1, respectively.IL-1β content were (1.44±0.47), (1.45± 0.23), and (1.61±0.33) ng??L-1 in low-, middle- and high-dose groups, respectively.TNF-α and IL-1β were (38.96±9.84) and (1.56±0.31) ng??L-1, respectively in Buchang Naoxintong group, (52.74±6.76) and (2.79±0.45) ng??L-1in the model control group, and (32.54±4.00) and (1.32±0.22) ng??L-1 in sham-operated group.TNF-αand IL-1βcontent were significantly higher in Buchang Naoxintong group than in sham-operated group ( P<0. 05 ) . TNF-α and IL-1β content were significantly decreased in Xifeng Tongnao high-, middle- and low-dose groups (all P<0.05). Conclusion Xifeng Tongnao capsules can protect brain tissue by supressing TNF-α and IL-1β and alleviating inflammatory injury from ischemia.

OBJECTIVETo explore the causative role of human bocavirus (HBOV) played in acute respiratory infection and diarrhea in children, a case-control study was prospectively conducted to investigate HBOV detection in symptomatic children with acute respiratory tract infection, diarrhea and asymptomatic children.

METHODBetween Oct. and Dec. of 2008, 436 nasopharyngeal aspirates (NPA) from hospitalized children with acute respiratory infection and 150 NPA from asymptomatic children undergoing cardiac operations were consecutively collected. During the same time, 220 stool samples were taken from outpatients with acute watery diarrhea and 200 control specimens were obtained from children without diarrhea. HBOV was screened in all samples by real-time PCR method. HBOV-positive respiratory samples were tested for other 9 common respiratory viruses and Mycoplasma pneumoniae. HBOV-positive fecal samples were also tested for common enteric viruses causing diarrhea.

RESULTHBOV was detected in NPA samples from 45 (10.3%) of 436 symptomatic patients and from 1(0.7%) of 150 asymptomatic control children. There was a statistically significant difference in the detection rates of HBOV between the symptomatic group and the asymptomatic group (P < 0.001). HBOV co-existence with other respiratory pathogens occurred in 44.7% (20/45) of NPA from symptomatic patients. HBOV was detected in 10.3% (43/417) children with community-acquired respiratory infection and 10.5% (2/19) children with nosocomial respiratory infection. Children with HBOV infection were 1.3 to 72 months of age (mean: 18.3 ± 13.6 months). HBOV was found positive in 6 (2.7%) of 220 stool samples from diarrheal outpatients and in 4 (2%) of 200 control samples. All children with HBOV positive detection in the stool samples were less than 4 years old. No statistical significance was found (P > 0.05) in HBOV between diarrhea patients and asymptomatic ones. In addition, 5 of 6 HBOV-positive fecal specimens from children with diarrhea were found co-infected with rotavirus.

CONCLUSIONThis study supports that HBOV is related to acute respiratory infection in children and HBOV infection usually occurs in infants and young children. However, further study is needed to clarify if HBOV plays a pathogenic role in diarrhea in children.

Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; Diarrhea ; virology ; Feces ; virology ; Female ; Human bocavirus ; isolation & purification ; Humans ; Infant ; Male ; Nasopharynx ; virology ; Parvoviridae Infections ; Prospective Studies ; Respiratory Tract Infections ; virology

BACKGROUND:Chondrocytes co-cultured with bone marrow stromal stem cells on the scaffold of platelet-rich plasma are found to proliferate, and besides proliferative growth, bone marrow stromal cells exhibit a tendency of differentiating into chondrocytes. OBJECTIVE:To study the effect of platelet-rich plasma and human umbilical cord mesenchymal stem cells (hUCMSCs) on cartilage repair. METHODS:Forty healthy New Zealand white rabbits were selected to establish models of cartilage defects, and then randomly divided into normal saline group, platelet-rich plasma group, hUCMSCs group and combination group. Platelet-rich plasma was prepared by using double centrifugations to prepare passage 3 hUCMSCs. After modeling, intra-articular injection of normal saline (0.5 mL), 12.5%platelet-rich plasma (0.5 mL), 1×107 hUCMSCs (0.5 mL), 12.5%platelet-rich plasma+1×107 hUCMSCs (total y 0.5 mL) was done in corresponding groups, respectively. After 12 weeks of modeling, the injured cartilage was grossly observed, and hematoxylin-eosin staining was used to observe cartilage repair under light microscope;according to the O'Driscol histologic standard, histological examination was performed. RESULTS AND CONCLUSION:The repair effect in the normal saline group was significantly better that in the platelet-rich plasma group, hUCMSCs group, combination group (P<0.05), while the platelet-rich plasma group and combination group also exhibit better outcomes than the hUCMSCs group (P<0.05). These findings indicate that both platelet-rich plasma and hUCMSCs can promote cartilage repair;moreover, platelet-rich plasma with or without hUCMSCs is superior to hUCMSCs alone in the cartilage repair.