Objective To investigate the therapeutic effect of immunodepressant on Crohn's disease (CD).Methods 105 patients with CD were collected from 1983 to 2006 in Peking Union Medical College Hospital.All of their clinical manifestations and therapeutic results were analyzed retrospectively.Results (1)The application of immunedepressant was significantly increased after the year of 2000 (34.7% vs 3.0%,P=0.000).(2)The application of immunodepressant and the clinical features of these patients were as fouows：①The number of patients with modcrate to severe CD were more than that with mild CD in those using immunodepressant (28.9% vs 0).②The use of immunodepressant was not related with the diseased site of CD.because there were no difference among the groups with lesion in small intestine (20.0%),colon(27.3%)and ileocolon(27.1%),P=0.726.③The serum albumin level of CD patients using immunodepressant was significantly lower than that of those not using(31.9 g/L vs 35.1 g/L,P=0.047).④The use of immunodepressant did not decrease the incidenee of operative treatment(38.5% vs 50.0%,P=0.320).(3)The rate of remission in 19 CD patients using azathioprine is 68.4%(13/19)and the percentage of neutrophil in the group with relief was lower than that without relief(0.76 vs 0.65,P=0.032).Conclusions Immunodepressant is playing more important role in the treatment of CD.The patients with moderate and severe CD with as well as lower serum albumin should be treated with immunodepressant as early as possible.Whether immunedepressant is necessary or not is not decided by the diseased site.CD patients with higher percentage of neutrophil have less therapeutic effect than those with lower.

Objective To investigate the alteration of serotonin-producing gastric enterochromaffin (EC)cell in patients with functional dyspepsia(FD).Methods Fifteen healthy volunteers and 33 patients with FD were enrolled.Proximal gastric mucosal EC cells were countered after immunohisto- chemistry staining.The ultrastructure of EC cell was observed by electromicroscope.Results The EC cells in proximal gastric mucosa in patients with FD were significant higher than that in controls(12.5?2.1 vs 8.3?1.4,t=2.353,P＜0.05),and the staining intensity of EC cell in patients with FD was also stronger than that in controls(3.72?0.42 vs 2.61?0.57,t=2.078,P＜0.05).The more sever the gastric mucosal inflammation was,the more number of EC cells and the stronger staining intensity were.Under the electromicroscopy,more Golgi apparatus,mitochondria and endoplasmic reticulum were found in EC cells.Special secreting particles were also found in cytoplasm.Conclusions EC cells may be involved in the pathogenesis of FD.The number of EC cell is related with the severity of gastric mucosal inflammation.

0.05).After the EN,the level of blood total protein,albumin and prealbumin significantly increased(P

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 find out the importance of human bocavirus (HBoV) as an infectious agent for population in Beijing, China. Seroprevalence study was conducted by using expressed recombinant major capsid VP2 protein as an antigen.Methods Serum specimens collected from infants and children who visited the Children's Hospital Affiliated to the Capital Institute of Pediatrics for health check-up and adults visiting the Xuanwu Hospital, Beijing for diseases other than respiratory infections from April 1996 to March 1997 were used for the investigation. The major capsid protein VP2 from HBoV was expressed in E. coli strain BL21 (DE3) with the transformed PET30b vector inserted with full-length VP2 gene of HBoV and the specific antigenicity of this expressed protein was validated by previous study. Western blotting was used to detect specific IgG antibody against HBoV in collected serum specimens diluted to 1:200. Mock expressed protein was E. coli cells strain BL21 (DE3) with the transformed PET30b vector without insert. Anti-His monoclonal antibody and rabbit anti-HBoV VP2 polypeptides hyper-immune serum were used as positive control for antibody detection.Results Out of 677 serum specimens tested, 400 (59.1% ) were positive for HBoV by Western blotting. About 45.3% (34/75) of the newborns under 1 month of age had anti-HBoV antibodies, and antibody positive rates were decreased in age groups of 1 and 2 months (41.4% and 31.3%, respectively) then increased in the following ages from 6 months to 7 years old ( from 45.6% to 69.7% ). The antibody positive rates were maintained at a relatively constant level ( about 70% ) in the age groups from 7 years to 40 years of age and became lower ( 61.8% - 62. 8% ) in those over 50 years.Conclusions The high seroprevalence of antibody against recombinant HBoV VP2 protein and early age antibody acquisition indicate that HBoV has been circulating in population of Beijing, China as early as in 1996 and most of children had been exposed to HBoV by the age of 7 years. Infants under the age of 6 months were susceptible to this virus.

Objective To analyze the gene expression profiles in relation to the migration ability of adult human bone marrow-derived neural stem cells (Md-NSCs), and identify the genetic basis of the high migration potential of Md-NSCs in the central nervous system (CNS). Methods Adult human bone marrow stromal celIs(BMSCs) obtained from adult healthy volunteers were induced to differentiate into Md-NSCs in vitro, and the expressions of the genes related to cell invasiveness and metastasis were investigated by microarray analysis. Quantitative real-time PCR (RT-PCR) was used to verify the microarray results. Results The results of microarray analysis revealed significant overexpressions of the genes MMP1, MMP2, MMP17, ITGA3, RhoB, RhoC and RhoD in the Md-NSCs as compared with the expressions in fresh normal human adult bone marrow cells depleted of red blood cells. Quantitative RT-PCR verified the overexpression ofMMP2 gene by 2.84×100 folds, ITGA3 gene by 2.22×102folds, and RhoC gene by 4.92×100 folds. Conclusion The high migration potential of the Md-NSCs in the CNS is probably associated with the overexpression of the genes that promote cell invasiveness and metastasis. These overexpressed genes are also important oncogenes, and therefore the tumorigenicity of the Md-NSCs warrants further investigation.

OBJECTIVEDiet is an important factor influencing blood pressure and, increases in dietary carbohydrate intake can raise blood pressure in adult rats. A previous study showed that the blood pressure of the rats fed with high-carbohydrate was 5-20 mmHg higher than that of control rats. While the mechanism involved is not clear. This study aimed to investigate the effects of high-sucrose intake on blood pressure of young Wistar rats and the role that sympathetic nerve system in the process.

METHODSMale neonatal Wistar rats were performed sympathectomy operation with 6-hydroxydopamine (6-OHDA) and then divided into four groups: (1) 0.1% VitC saline-common diet group (VN), (2) 0.1% VitC saline-high sucrose (VS), (3) 6-OHDA-common diet group (OHN) and (4) 6-OHDA-high sucrose (OHS) after three week. The data on the body weight (BW), systolic blood pressure (SBP) and heart rate (HR) were recorded. Then the level of blood glucose, serum insulin and angiotensin II (AngII) were measured and the functional studies of the thoracic aorta was performed.

RESULTSThe VS group exhibited higher SBP than the OHS group from the 6th week (113.7 +/- 4.2 mmHg vs. 104.0 +/- 5.8 mmHg, P < 0.01) and the VN group from the 7th week (117.6 +/- 6.3 mmHg vs. 109.6 +/- 4.6 mmHg, P < 0.01), while the SBP of the VN group was similar to those of the OHN group and the OHS group (P > 0.05). No significant differences in blood glucose, serum insulin and insulin sensitive index (ISI) were found among the four groups. The thoracic aorta segments of the VS group had higher contractive response to AngII (P < 0.01) and NE (P < 0.05) than the VN group, but the relaxations to acetylcholine (ACh) and nitroglycerine (NTG) showed no difference among the four groups (P > 0.05).

CONCLUSIONThe high-sucrose diet might elevate the blood pressure in young Wistar rats and the sympathetic system may play an important role in this process.

Angiotensin II ; blood ; Animal Feed ; Animals ; Blood Glucose ; metabolism ; Blood Pressure ; drug effects ; physiology ; Body Weight ; drug effects ; Dietary Sucrose ; administration & dosage ; Insulin ; blood ; Male ; Oxidopamine ; administration & dosage ; blood ; Rats ; Rats, Wistar ; Sympathectomy

The aim of this study was to characterize the N and E protein encoding genes of a new human coronavirus (HCoV-NL63) which was identified from one of the clinical specimens (BJ8081) collected from a 12 years-old patient with acute respiratory infection in Beijing. The complete N and E gene sequences of HCoV-NL63 were amplified from clinical sample by RT-PCR, then were cloned into the pCF-T and pUCm-T vectors respectively and sequenced. The complete sequences of N and E genes were submitted to GenBank by Sequin and compared with N and E genes of prototype HCoV-NL63 and the other coronaviruses published in GenBank. The secondary structure and the characteristics of sample BJ8081 N and E proteins were predicted by bioinformatics. It was indicated that the N and E genes amplified from sample BJ8081 were 1134 bp and 234 bp in length and the predicted proteins including 377 amino acids and 77 amino acids, respectively. The data suggested that the region of amino acids 78-85 within N protein probably was the conserved region for all coronaviruses identified so far including HCoV-NL63. The region of amino acids 15-37 for E protein was probably the transmembrane domain. In conclusion, the recombinant plasmids pCF-T-8081 N and pUCm-T-8081 E were successfully constructed and sequenced, and the data predicted by bioinformatics are helpful for the further analysis of HCoV-NL63.
Amino Acid Sequence ; Child ; China ; Computational Biology ; methods ; Coronavirus ; classification ; genetics ; metabolism ; Coronavirus Infections ; virology ; Humans ; Molecular Sequence Data ; Nucleocapsid Proteins ; chemistry ; genetics ; metabolism ; Phylogeny ; Protein Structure, Secondary ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Viral Envelope Proteins ; chemistry ; genetics ; metabolism

OBJECTIVETo establish a rapid, sensitive and specific reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for detecting human respiratory syncytial virus (RSV) in respiratory samples collected from children with acute respiratory infections.

METHODAccording to the conserved matrix gene sequences of respiratory syncytial virus subtypes A and B downloaded from GenBank, primers were designed and RT-LAMP assay was developed to detect RNA of RSV sensitivity of the RT-LAMP method was evaluated by using ten-fold serially diluted in vitro-transcribed matrix RNA fragments from RSV A and RSV B, respectively. Specificity of the RT-LAMP method was tested through cross-reaction with other RNA and DNA viruses. Then 5 RSV strains isolated from clinical specimens using tissue cultures were tested by RT-LAMP assay. A total of 101 nasopharyngeal aspirates from hospitalized patients with acute respiratory infections which had been tested by direct immunofluorescence assay (DFA), including 40 positive for RSV and 61 negative for RSV, were tested by RT-LAMP assay and by RT-nested PCR.

RESULTSensitivity analysis indicated that this RT-LAMP method was able to detect 1 copy/µl of RSV A and RSV B RNA, no amplification was shown in RT-LAMP with DNA or cDNA from other viruses in 60 min, revealed that the RT-LAMP assay is highly specific. Five RSV isolates confirmed as 4 RSV A and 1 RSV B previously were detected by RT-LAMP method as positive in 30 min. For those 101 specimens tested, 37 were RSV positive determined by RT-LAMP assay, as well as 35 RSV positive by RT-nested PCR. The total coincidence rate of RT-LAMP assay with DFA and RT-nested PCR in detecting RSV is 95.0%, 94.1% with Kappa value 0.895 and 0.871, respectively.

CONCLUSIONA new, sensitive, accurate and rapid method, RT-LAMP assay for detecting human respiratory syncytial viruses from nasopharyngeal aspirates was developed, which should be helpful in rapid detection of RSV from respiratory tract samples of children.

Acute Disease ; Child ; Child, Preschool ; DNA Primers ; Humans ; Infant ; Molecular Diagnostic Techniques ; Nasopharynx ; virology ; Nucleic Acid Amplification Techniques ; RNA, Viral ; isolation & purification ; Respiratory Syncytial Virus Infections ; diagnosis ; Respiratory Syncytial Virus, Human ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity

OBJECTIVETo understand the impact of human parainfluenza virus (HPIV) on acute respiratory infections in infants and young children in Beijing.

METHODSMultiplex reverse transcription-PCR was used to amplify the hemagglutinin (HA) gene fragment of HPIV from clinical specimens. Primer pairs derived from a conserved region of the HA genes of HPIV were used to develop the multiplex RT-PCR for detecting and typing HPIV. The sensitivity and specificity of the method were determined by using various RNA and DNA viruses as controls. Specimens collected from 3519 children with acute respiratory infections from Aug. 2003 to Apr. 2006 were analyzed for HPIV by the multiplex RT-PCR as well as for other respiratory viruses by virus isolation and/or indirect immunofluorescent assay (IFA). Ten amplicons with expected molecular weight matching different types of HPIV were randomly selected for sequence analysis.

RESULTSOnly the cDNA from the isolated strains of HPIV 1 and 3 was positive by the multiplex RT-PCR. Phylogenetic analysis for those 10 amplicons' sequences which belong to HPIV 1 - 4 types respectively as determined by multiplex-PCR indicated that these specimens were truly HPIV positive. These 10 HPIV positive specimens included two specimens of type 4 which was further subtyped as HPIV4A and 4B by sequence analysis. With the multiplex RT-PCR, HPIV were detected in 349 out of 3519 specimens with the positive rate of 9.9% (349/3519), which is higher than 4.8% by the methods of virus isolation and/or IFA. And the HPIV positive rates were high in patients with not only acute upper but also lower respiratory tract infection. No regular seasonality distribution of HPIV infection was found. HPIV 1 and 3 were more common than HPIV 2 and 4.

CONCLUSIONWith higher sensitivity and specificity than virus isolation and IFA, multiplex RT-PCR is beneficial for the etiologic and epidemiologic studies on HPIV, as well as for HPIV typing. The data from this study indicate that HPIV is one of the important etiological viruses of acute respiratory tract infections in infants and young children in Beijing.

Child, Preschool ; China ; epidemiology ; Genes, Viral ; HN Protein ; genetics ; Humans ; Infant ; Paramyxoviridae Infections ; epidemiology ; virology ; Phylogeny ; Prevalence ; Respiratory Tract Infections ; epidemiology ; virology ; Respirovirus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity