OBJECTIVETo prepare anti-recombinant protein antibody from immunized mice with recombinant nucleocapsid protein (NP) of human influenza A3 (IFV-A3) virus expressed in prokaryotic cell, and to explore the feasibility of utilizing anti-recombinant protein antibody to detect influenza A virus.

METHODSNP genes of human influenza A virus were analyzed with computer softwares of ClustalX, Antheprot, et al. to determine the antigenicity in conserved regions. Three different partial NP genes were harvested and cloned into pET-28(c) plasmid, the recombinant plasmids were induced to express partial NP segments in BL21 cells. The recombinant proteins were purified with Ni-agarose by affinity chromatography and immunized BALB/c mice. The polyclonal antisera harvested from mice were analyzed with Western Blot and immunohistochemistry assays to detect the reactions with IFV-A.

RESULTSThree recombinant plasmids were expressed with high yield in BL21 cells, about 15-20 mg/L. Western Blot results indicated that the three prepared antisera (1:2000) positively reacted with NP from IFV-A3-infected cells. And immunohistochemistry assays suggested that anti-NP1, anti-NP2, anti-NP3 antisera positively reacted with IFV-A3 or IFV-A1-infected MDCK cells, with titers of 1:640 to 1:1280.

CONCLUSIONThe recombinant NP of IFV-A3 would induce polyclonal antibodies with high titers in mice. The polyclonal antibodies would cross-react with IFV-A3 and IFV-A1. It is feasible to predict the antigenicity with systematical bioinformatics analyses and then induce anti-IFV antibodies with high dilutions, and it is possible to be utilized in the early detection and subtyping analyses of IFV-infections.

Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Influenza A virus ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology

Brain Neoplasms ; Germinoma ; Humans ; Pineal Gland

This study aims to analyze the molecular epidemiological characteristics of HIV-1 strains prevailing among men who have sex with men (MSM) in Beijing, China. The pol gene fragments from 250 newly diagnosed HIV-1-infected MSM individuals during 2006-2010 in Beijing were amplified by RT-nested PCR, sequenced, and phylogenetically analyzed. HIV-1 pol gene from 189 individuals were amplified and analyzed; 81 (42. 9%), 3 (1. 6%), 2 (1.0%), 88 (46. 6%), and 15 (7.9%) individuals were infected with HIV-1 subtypes B, B', C, CRF01_AE, and CRF07_BC, respectively. The subtypes B and CRF01_AE could both be grouped into two clusters, and CRFO7_BC strains shared high homology and were presumed to originate from a common ancestor. The HIV-1 circulating in MSM in Beijing had a lower genetic diversity than in heterosexuals. The HIV-1 epidemic (2006-2010) in MSM in Beijing was actually a rapid spread of HIV-1 CRF01 AE and B, or rather native strains of the two viruses.
Adult ; China ; epidemiology ; Epidemics ; Genetic Variation ; HIV Infections ; diagnosis ; epidemiology ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Homosexuality, Male ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; Young Adult

Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Rickets ; diagnosis ; etiology ; therapy ; Treatment Outcome ; Vitamin D Deficiency ; complications

OBJECTIVETo study the MRI manifestation of testicular tumor and the value of MRI in the diagnosis of the disease.

METHODSWe retrospectively analyzed 23 cases of pathologically confirmed testicular tumor, and observed the morphological characteristics, signals and surrounding conditions of the tumor using plain and enhanced MRI scanning.

RESULTSOf the 23 cases, seminoma was identified in 7, mixed germinoma in 3, teratoma in 3, endodermal sinus tumor in 2, epidermoid in 1, Leydig cell tumor in 1, leucoma in 1, nonspecific inflammatory mass in 3, and tuberculosis in 2. MRI revealed the precise locations and specific characteristics of

CONCLUSIONBased on MRI findings and clinical manifestation, most testicular tumors can be diagnosed correctly.

Adolescent ; Adult ; Carcinoma, Embryonal ; diagnosis ; Child ; Child, Preschool ; Endodermal Sinus Tumor ; diagnosis ; Germinoma ; Humans ; Infant ; Leydig Cell Tumor ; diagnosis ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Seminoma ; diagnosis ; Teratoma ; diagnosis ; Testicular Neoplasms ; diagnosis ; Young Adult

OBJECTIVETo investigate the relationships between the severity of childhood allergic rhinitis (AR) and the peripheral blood eosinophil count, serum eosinophil cationic protein (ECP), total IgE (tIgE), and allergen-specific IgE (sIgE) levels.

METHODSA total of 138 children with AR aged 3 to 17 (9.96 ± 3.78, x() ± s) years old were enrolled in the study. All children had persistent AR sensitized to house dust mites with a clinical history of 3 months to 12 (4.21 ± 2.72) years. The disease severity was evaluated using 10 cm-visual analogue scale (VAS), and the serum levels of ECP, tIgE and sIgE were determined using an ImmunoCAP system. Statistical analysis was conducted with SPSS11.0 software.

RESULTSAmong 138 children with AR, the VAS scores for global severity of rhinitis and nasal obstruction symptom were 5.32 ± 2.16 and 4.78 ± 2.45, respectively. Blood eosinophil count was 0.39 [0.24; 0.63] (M[P(25); P(75)]) ×10(9)/ml. Serum levels of ECP and total IgE were 10.60 [3.26; 30.80] µg/L and (2.50 ± 0.53) log kU/L, respectively. Serum levels of allergen-sIgE against Dermatophagoides pteronyssinus and Dermatophagoides farinae were 58.20[24.75; > 100] kUA/L and 54.95 [24.55; > 100] kUA/L, respectively. The VAS scores of nasal obstruction symptom, but not global severity of rhinitis, were positively related to the duration of AR (r = 0.215, P = 0.011) and the levels of serum ECP (r = 0.196, P = 0.022) in bivariate correlation analysis. There was also a significant correlation between the serum ECP level and the blood eosinophil count (r = 0.295, P = 0.000). No relationships of blood eosinophil count, and serum tIgE and sIgE levels with global severity of rhinitis as well as nasal obstruction symptom were found (all P > 0.05).

CONCLUSIONSThese results suggested that the severity of nasal obstruction was positively correlated with the duration of rhinitis and the levels of serum ECP in childhood persistent AR due to house dust mites, indicating the disease severity might be related to chronic inflammatory process.

Adolescent ; Animals ; Child ; Child, Preschool ; Eosinophil Cationic Protein ; blood ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Pyroglyphidae ; immunology ; Retrospective Studies ; Rhinitis, Allergic ; Rhinitis, Allergic, Perennial ; blood ; immunology

OBJECTIVEUsing molecular epidemiology method to characterize human immunodeficiency virus type 1 (HIV-1) subtype CRF01 _ AE strains being prevailed in Zhejiang province.

METHODSGag fragments of the HIV-1 strains were amplified by nested-polymerase chain reaction (nPCR) from the DNA extracted from whole blood of HIV-1 infected individuals in Zhejiang province. PCR products were sequenced and analyzed by phylogenetic method.

RESULTS81 HIV-1 subtype CRF01 _ AE sequences were identified from the 192 samples that sequenced successfully. As one of the dominant subtypes in Zhejiang, CRF01 _ AE was transmitted mainly by heterosexual or homosexual contact in local residents. In migrants living in Zhejiang, CRF01 _ AE were transmitted mainly by heterosexual contact or injecting drug use. There were three main clusters in the phylogenetic tree which bootstrap value was larger than 60. We named the clusters with group MIX (47 sequences), group SEX (7 sequences) and group MSM (12 sequences) based on the transmission. Pairwise DNA distances in the gag region within the three groups and between CM240 were different (P = 0.000). Data through the analyses of deduced amino acid sequences from the three groups showed that several signature amino acid sites were distinct from the same positions of the subtype reference strains.

CONCLUSIONThe CRF01 _ AE strain prevailing in Zhejiang province was from several sources, transmitted by more than three different transmission routes, and becoming the main subtypes circulating in homosexual population in this study. More attention needs to be paid to the epidemic characteristic of CRF01 _ AE.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; methods ; Phylogeny ; Young Adult

BACKGROUNDThe delayed diagnosis of pelvi-ureteric junction (PUJ) disruption in children following blunt abdominal trauma can result in loss of function of the involved kidney. We examined the potential for kidney preservation and the limits of diagnostic delays.

METHODSA retrospective review of 17 cases of PUJ disruption at Beijing Children's Hospital from 1993 to 2009 was done with respect to diagnosis, treatment and follow-up.

RESULTSThe interval from trauma to diagnosis of PUJ disruption was (52 ± 52) days. If one case with nephrectomy was excluded, the interval from trauma to diagnosis was (40 ± 20) days. The average time between injury and first treatment was (49 ± 25) days. Pelvi-ureteric reanastomosis and caliceal ureterostomy were performed separately in 11 and 4 patients, respectively. Ileal replacement for ureter injuries was finally performed in one patient. Hydronephrosis of the injured kidney was reduced and the function improved in 15 out of 17 patients (88%). Only one patient received nephrectomy and the nephrectomy rate was 5.9%.

CONCLUSIONDifferential renal function at the PUJ disruption side can be saved and the rate of nephrectomy reduced by appropriate surgery if the time to diagnosis and first treatment is limited to within two months.

Abdominal Injuries ; complications ; surgery ; Child ; Child, Preschool ; Female ; Humans ; Kidney ; injuries ; surgery ; Kidney Pelvis ; injuries ; surgery ; Male ; Retrospective Studies ; Ureter ; injuries ; surgery ; Ureteral Obstruction ; etiology ; surgery

To explore the genetic characteristics of viral quasispecies in HIV-1 CRF07_BC infections among intravenous drug users (IDU), the gp120 fragments of HIV-1 env gene were amplified from plasma samples collected from 6 CRF07_BC infected persons using single genome amplification and sequencing (SGA/ SGS) method, and 11 to 28 sequences were obtained from these samples, respectively, A neighbor-joining phylogenetic tree was reconstructed to describe the genetic characteristics of viral quasispecies. The Simplot, segments' phylogenetic trees and diversity plots based on average pairwise distance (APD) were used to identify the recombination events between quasispecies. The SGA sequences derived from single specimen formed a large monophyletic cluster in the neighbor-joining phylogenetic tree and showed the complex topologic structures of viral quasispecies. Of the 6 CRF07_BC infected patients, only one possessed the high genetic homogeneity, whereas the other five individuals showed high heterogeneity, with two to four subclusters inside the monophyletic cluster for each specimen. In addition, the recombinant events were identified among viral quasispecies from 3 cases. The results show SGA technique and phylogenetic analyses are useful tool to investigate the intrahost CRF07_BC gp120 complex quasispecies variation and high genetic diversity.
Adult ; Drug Users ; Female ; HIV Infections ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Humans ; Male ; Molecular Sequence Data ; Phylogeny ; Substance Abuse, Intravenous ; virology ; Young Adult

The investigation on Salvia przewalskii Maxim was carried out to find the relationship of the constituents and their pharmacological activities. The isolation and purification were performed by various chromatographies such as silica gel, Sephadex LH-20, RP-C18 column chromatography, etc. Further investigation on the fraction of the 95% ethanol extract of Salvia przewalskii Maxim yielded przewalskin Y-1 (1), anhydride of tanshinone-II A (2), sugiol (3), epicryptoacetalide (4), cryptoacetalide (5), arucadiol (6), 1-dehydromiltirone (7), miltirone (8), cryptotanshinone (9), tanshinone II A (10) and isotanshinone-I (11). Their structures were elucidated by the spectral analysis such as NMR (Nuclear Magnetic Resonance) and MS (Mass Spectrometry). Compound 1 is a new compound. Compounds 4 and 5 are mirror isomers (1 : 3). Compounds 4, 5, 6, 8, 11 were isolated from Salvia przewalskii Maxim for the first time.
Diterpenes ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Phenanthrenes ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Salvia ; chemistry