OBJECTIVETo study the human myxovirus resistant protein A (MxA), a specifically induced peptide by interferon I, and to use its level as a diagnostic criterion for viral infections.

METHODSAnti-MxA antisera from immunized mice were prepared with the expressed MxA protein of pET32a-MxA in E. coli BL-21(DE3). To confirm the antiserum activity and specificity, the expression product of BL21, wild type MxA pEGFP-C1-wMxA and site-directed mutant MxA pEGFP-C1-mMxA(N589S) stably transfected 3T3 cells and induced A549 cells were detected by Western blot with the antisera using non-MxA transfected or non-IFN-beta induced cells, intact A549, NIH 3T3 cells transfected with pEGFP-C1 and pET32a (+)-transformed BL-21 as controls.

RESULTSThe antisera had specific positive immunoreactivity to the NIH3T3 cells transformed with pEGFP-C1-wMxA and pEGFP-C1-mMxA, INF-beta induced A549 cells and BL21 proteins expressed with pET32a (+)-MxA. The hybridization signals from IFN-beta induced A549 cells depended on the IFN-beta inducing concentrations. Meanwhile, immunohistochemical assay showed that NIH 3T3 cells with pEGFP-C1-wMxA and pEGFP-C1-mMxA had > 98% of positive cells at 1:50 dilution of the serum and A549 cells induced by 20 ng/mL IFN-beta for 48 h showed 95% positive cells. pEGFP-C1-transfected NIH 3T3 cells were all negative.

CONCLUSIONAnti-sera are highly specific to diversified MxAs. The antibody is detectable by Western blot, immunocytochemistry and immunofluorescence assay.

Animals ; Antibody Specificity ; Cell Line, Tumor ; GTP-Binding Proteins ; genetics ; immunology ; metabolism ; Gene Expression Regulation ; Humans ; Mice ; Myxovirus Resistance Proteins ; NIH 3T3 Cells ; Species Specificity

OBJECTIVELate onset neonatal septicemia (systemic infection after 72 hours of life) remains a major cause of neonatal morbidity and mortality. Early treatment with appropriate antibiotics is critical since infected infants can deteriorate rapidly. The aim of this study was to review the pathogens responsible for late onset neonatal septicemia (LONS) and their antimicrobial susceptibilities in order to guide the initial selection of appropriate antibiotics for infants with suspected LONS.

METHODSA retrospective chart review of all cases with LONS seen in the neonatal intensive care unit (NICU) of Yuying Children's Hospital of Wenzhou Medical College from January 1, 2002 to December 31, 2005 was conducted. All cases were selected based on the clinical presentation and at least one positive result of blood culture. The basic clinical characteristics and the results of blood culture and antimicrobial susceptibilities were analyzed.

RESULTSA total of 102 cases with LONS were identified. Among those 102 cases, 80 were community acquired (infants admitted from home and the blood culture was done on admission) and 22 were hospital acquired (infants became sick while in the NICU and the blood culture was done prior to use of antibiotics). The clinical presentations were non-specific. Compared to the infants with community acquired LONS, infants with hospital acquired LONS were usually born more prematurely (mean gestational age 33 +/- 3 vs 39 +/- 2 wks, t = 2.255, P < 0.01), with lower weight (mean weight 1.79 +/- 0.70 vs 3.23 +/- 0.67 kg, t = 8.818, P < 0.01) and with younger age (mean age 12 +/- 6 vs 16 +/- 7 days, t = 7.581, P < 0.05). Of the 102 cases, a total of 103 strains of bacteria were isolated. Among the pathogenic bacteria isolated, the most common were coagulase-negative Staphylococcus (CoNS) (50/103, 48.5%), followed by Klebsiella pneumoniae (16/103, 15.5%). The main pathogens for community acquired LONS were Staphylococcus species and Escherichia coli. The most important pathogen responsible for hospital acquired LONS was Klebsiella pneumoniae. Most (> 80%) of the Staphylococcus especially CoNS were resistant to common antibiotics such as penicillin, erythromycin and cefazolin. Significant numbers (6/9) of Staphylococcus aureus isolated were methicillin-resistant Staphylococcus aureus (MRSA). However, all of the Staphyloccus isolates were sensitive to vancomycin. Almost all (15/16) of the Klebsiella pneumoniae isolated were multi-drug resistant due to production of extended-spectrum beta-lactamases (ESBLs). They were sensitive only to a few antibiotics such as carbapenems, aminoglycosides and quinolones. There was also one strain of vancomycin-resistant Enterococcus (VRE). Furthermore, there was no a single case of late onset neonatal sepsis due to infection with group B Streptococcus (GBS).

CONCLUSIONSThe clinical manifestations of late onset neonatal sepsis are usually non-specific. GBS is not a significant pathogen responsible for community acquired LONS in the Wenzhou area. There are increasing numbers of multi-drug resistant bacterial species isolated from the newborn infants with late onset neonatal septicemia, which is most likely due the non-restricted use of antibiotics in the hospitals as well as in the communities. A routine blood culture should be taken from any newborn infant who is suspected of LONS and empirical use of appropriate antibiotics should be initiated as soon as the blood specimen for culture has been drawn. To reduce the occurrence of multi-drug resistant bacteria, the use of antibiotics especially the third generation cephalosporins in neonates should be restricted as much as possible.

Community-Acquired Infections ; microbiology ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Drug Resistance, Multiple, Bacterial ; Female ; Humans ; Infant, Newborn ; Male ; Retrospective Studies ; Sepsis ; microbiology

China ; Genotype ; Measles virus ; genetics ; isolation & purification ; Molecular Epidemiology ; methods ; Nucleoproteins ; genetics ; Polymerase Chain Reaction

BACKGROUNDPulmonary embolism (PE) is rare and seldom considered in adolescent patients; however it occurs with a greater frequency than is generally recognized, and it is a potentially fatal condition. The aim of the current study was to understand its epidemiology, clinical features and the cause of delay of its diagnosis in adolescents.

METHODSA retrospective analysis of nine adolescents with acute PE admitted to the Peking University Third Hospital over the past 16-year period was performed. The epidemiology, clinical features and risk factors of the adolescents were described and compared with those of adults and elderly patients. The time to diagnosis and misdiagnosed diseases were analyzed. Pretest probability of PE was assessed retrospectively by the Wells score and revised Geneva score.

RESULTSThe incidence of PE was 43.6 per 100 000 hospitalized adolescents in our hospital. The incidence of PE in adolescents was much lower than that in adults and PE is diagnosed in about 1/50 of elderly people. The clinical features in adolescents were similar to those in adults. But fever and chest pain were more common in adolescents (P < 0.05). The major risk factors included surgery, systemic lupus erythematosus (SLE), thrombocytopenia, long-term oral glucocorticoids and trauma. The mean diagnostic time was (7.8 ± 8.4) days. Six cases had a delayed diagnosis. The mean delay time from symptom onset to diagnosis was (11.0 ± 8.8) days. The time of presentation to diagnosis in patients initially admitted to the emergency department was less than one day, and was much shorter than the time in outpatients, (9.4 ± 7.5) days. Most of the patients were initially misdiagnosed with a respiratory tract infection. Most patients' values of Wells score or revised Geneva score were in the moderate or high clinical probability categories; 88% by Well score vs. 100% by revised Geneva score.

CONCLUSIONSPE was seldom considered in the adolescent patients by physicians, especially outpatient physicians, so the diagnosis was often delayed. If adolescent patients complain of dyspnea or chest pain or syncope with/without fever, and they had risk factors such as surgery, thrombocytopenia and trauma, PE should be considered and included in the differential diagnosis.

Adolescent ; Adult ; Diagnosis, Differential ; Diagnostic Errors ; Humans ; Male ; Probability ; Pulmonary Embolism ; diagnosis ; epidemiology ; etiology ; Retrospective Studies ; Risk Factors

To identify HLA novel allele in Chinese Han individuals, an unknown HLA-A allele was detected by PCR-SSP and FLOW-SSO in Chinese Han individuals. Heterozygous sequence-based typing (SBT) showed that there were 3 differences compared with database in exon 2. Its anomalous patterns suggested the possible presence of either a novel A * 30 or a novel A * 24. To separate the two alleles and to determine whether the allele is novel, the HLA-A * 30 and HLA-A * 24 alleles were amplified separately by using a commercial kit for the single allele-specific sequencing strategy, and both alleles for exons 2 - 4 were sequenced according to the manufacturer' protocol. To prepare B-lymphoblastoid cell line of the novel HLA allele by using Epstein-Barr virus-infected B-lymphoblastoid cells in the peripheral blood. The results indicated that the sequencing results showed HLA-A alleles of the sample to be HLA-A * 240201 and a new A * 30 allele. The sequences of the new A*30 were identical to those of HLA-A * 300101 except for three nucleotide changes in exon 2: at nt 121 (A-->C), nt 123 (T-->C) and nt 126 (A-->G), resulting in an amino acid residue substitution from S (AGT) to R (CGC) at codon 17 and a synonymous substitution from G (GGA) to G (GGG) at codon 18. Immortalized B-lymphoblastoid cell line of the novel HLA-A * 3018 allele was achieved, the sequence of HLA-A * 3018 allele was submitted to GenBank and its accession number was DQ872509. In conclusion, the HLA-A * 3018 is a novel HLA-A allele and has been officially named HLA-A * 3018 by the WHO Nomenclature committee in August 2006 (HWS10004039).
Alleles ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; HLA-A Antigens ; genetics ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA

Objective To evaluate the chemopreventive effects of boswellic acid and curcumin on 7,12-dimethyl benzanthracene (DMBA)-induced oral carcinogenesis in the hamster cheek pouch model.Methods Male Syrian golden hamsters(6-8 weeks old,80-130 g in weight) were randomly divided into seven groups,with group A serving as the untreated negative control.The left cheek pouch of the remaining hamsters was topically treated with 0.5％ DMBA in mineral oil three times a week for 6 weeks.They were then randomized to six groups with group B serving as a positive control and receiving no further treatment.Groups C-G were treated topically with 5,10 mg/L boswellic acid,5,10 μ moL/L curcumin,or the combination of 5 mg/L boswellic acid and 5 μmnol/L curcumin three times per week for 18 weeks.The animals were injected with bromodeoxyuridine intraperitoneally at 50 mg/kg 2 h prior to killing.At the 25 th week all the hamsters were sacrificed and cheek pouch tissue was harvested.One half of the tissue was snap frozen in liquid nitrogen for analysis of arachidonic acid metabolites,and the other half was fixed in 10％ phosphate-buffered saline(PBS)-buffered formalin for histopathological examination.Results Six-weeks of DMBA followed by 18-weeks of topical application of boswellic acid and curcumin,both boswellic acid (5,10 mg/L) and curcumin (5,10 μmol/L) significantly inhibited the incidence from 93.8％ to 73.9％ (P＞0.05),numbers from 2.19 ± 0.98 to 1.13 ± 0.81 (P ＜ 0.01) and size of visible tumors.Microscopically the incidence of squamous cell carcinoma and BrdU index were also significantly suppressed by boswellic acid and curcumin.Conclusions Both boswellic acid and curcumin were effective in preventing oral carcinogenesis in DMBA-induced hamster cheek pouch model.

OBJECTIVETo determine the impact of passive smoking and the protective effect of antioxidants such as vitamin E and quercetin on learning and memory ability of mouse offsprings.

METHODSA passive smoking model of pregnant mice was established. Learning and memory ability was evaluated by the water maze test and long term potentiation (LTP). Nitric oxide (NO), content, nitric oxide synthase (NOS), acetylcholinesteras (Ache) activity in brain, vitamin E concentration, and reactive oxygen species (ROS) in serum were determined. The latency period (the time during which the mice swim from the starting position to the ending position) and errors (the number of mice entering the blind end) in control and antioxidant intervention groups were compared with those in the smoke exposure group after 6 days.

RESULTSThe latency period as well as errors in the air, control diet, tobacco smoke (TS), and vitamin E diet groups were decreased significantly as compared with the TS and control diet groups (P<0.05). LTP was restrained in the TS and control diet groups. LTP in all the antioxidant diet groups was significantly increased compared with the TS and control diet groups. In addition, NOS and acetylcholinesteras (Ache) activitiy was significantly higher in the TS and control diet groups than in the air and control diet group. NO content was not significantly different among the different groups, and significantly lower in the TS and vitamin E diet groups than in the TS group, control diet group, quercetin diet group, and mixture diet group (P<0.05). Vitamin E concentration and ROS activity in serum were correlated with the outcome of water maze and LTP.

CONCLUSIONPassive smoking reduces LTP formation by disturbing the hippocampus function of mice, by decreasing NOS and Ache activity and increasing NO content. Antioxidants (especially vitamin E) partially improve the learning and memory ability of offsprings whose mothers are exposed to tobacco smoke during pregnancy.

Animals ; Antioxidants ; administration & dosage ; Body Weight ; Brain ; enzymology ; metabolism ; Female ; Learning ; Long-Term Potentiation ; Male ; Maternal Exposure ; Maze Learning ; Memory ; Mice ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Pregnancy ; Tobacco Smoke Pollution

Aged ; Coronary Angiography ; Endocardium ; pathology ; Heart Aneurysm ; diagnosis ; diagnostic imaging ; Humans ; Male ; Myocardial Infarction ; complications

OBJECTIVETo investigate the regulatory effect of the nonsteroidal anti-inflammatory drug NS398 on the expression of the RECK gene in the animal model of prostate cancer.

METHODSNude mouse models of prostate cancer were divided into an experimental and a control group, the former fed with NS398 at 0.1 mg/g per day for 10, 20 and 30 days, while latter left without medication. All the mice were killed at 30 days, the mRNA expressions of RECK and MMP-9 in the tumor tissues measured by RT-PCR, and the protein level of RECK evaluated by Western blot.

RESULTSBoth the mRNA and protein expressions of RECK were increased, while the level of MMP-9 decreased, in an obviously time-dependent manner in the experimental group as compared with the control.

CONCLUSIONNS398 obviously inhibits the pathogenesis and metastasis of prostate cancer, which may be attributed to its induction of the expression of the RECK gene and suppression of the expression of MMP-9.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; GPI-Linked Proteins ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Nude ; Nitrobenzenes ; pharmacology ; Sulfonamides ; pharmacology ; Tumor Cells, Cultured