OBJECTIVETo study the role of the HBV-infected mothers' PBMC in intrauterine transmission of HBV to their fetuses.

METHODSThirty pregnant women with serum HBV DNA negative and PBMC HBV DNA positive and their newborns were used as the study group. Ten pregnant women with serum HBV negative and their infants served as the control group. HBV DNA in serum and in PBMC was detected using nested polymerase chain reaction (n-PCR). The mothers' PBMC in newborns' peripheral blood was examined using heminested-PCR.

RESULTSFour newborns were serum HBV DNA positive and 8 newborns were HBV DNA positive in PBMC in the study group. Among them, 2 newborns were HBV DNA positive in both serum and PBMC, 6 cases were positive in PBMC only, and 2 cases were positive in serum only. Five mothers had the GSTM1 gene; and it was not detected in 3 newborns. Among the 8 newborns with HBV DNA positive in PBMC, 3 did not have the GSTM1 gene, at the same time their mothers possessed the GSTM1 gene. Mothers' PBMC were detected in all of these three newborns' peripheral blood. HBV DNA in serum and in PBMC of the control group infants were all negative.

CONCLUSIONHBV-infected PBMC of the mother may serve as a vector in HBV intrauterine infection.

Adult ; DNA, Viral ; analysis ; Female ; Hepatitis B virus ; isolation & purification ; Hepatitis B, Chronic ; blood ; transmission ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Leukocytes, Mononuclear ; virology ; Pregnancy ; Pregnancy Complications, Infectious ; virology

Objective By detecting the distribution and homology analysis of Acinetobacter baumanii in the ICU ward,to con-trol nosocomial infection,provide theoretical basis to take effective measures.Methods From January 2016,20 patients in ICU of Shaanxi Provincial People's Hospital were taken into group.The samples of sputum and surrounding environment samples were cultured.The homology of Acinetobacter baumannii was analyzed by MALDI-Biotyper software,antimicrobial susceptibility test was analyzed by K-B.Results 27 strains of Acinetobacter baumannii were detected,mainly comes from sputum,hands of medical staffs and the environment,homology analysis results showed that the 27 strains of Acinetobacter baumannii was divided into two clusters(Ⅰtype 1 1 strains,Ⅱ type 1 6 strains),most of Acinetobacter baumannii were multi-resistant bacteria,except for the polymyxin B,minocycline and SCF.Conclusion The ways of transmission of Acineto-bacter baumanii in ICU were by medical personnel hand,pollution of the medical equipment and so on,strengthening the dis-infection and reasonable application of antimicrobial agents were taken advantageous for the prevention and control of acine-tobacter baumannii infection and transmission.

This study was aimed to establish ELISA for recombinant bovine IFN-gamma (BovIFN-gamma) detection and provide a new method for diagnosis of pathogenic infection. The total RNA was isolated from peripheral blood leucocytes cultured with PHA mitogen stimulation. Then bovine IFN-gamma (BovIFN-gamma) gene cDNA was amplified by RT-PCR and cloned into pET28a to obtain the expression plasmid designated as pETBovIFN-gamma. The pETBovlFN-gamma was further transformed into competent E. coli BL21 cells and a 18kD His-tagged protein as expected was expressed after IPTG induction. By using purified recombinant BovIFN-gamma as antigen and lymphocyte-hybridoma technique, four hybridoma cell lines which stably secreted monoclonal antibodies against rBovIFN-gamma were generated, designated as A7, A10, G6, and G10. The immunoglobin subset was identified as IgG1 . Western-blotting analysis and ELISA demonstrated that the monoclonal antibodies secreted by all the four hybridoma cell lines could react specifically to the recombinant BovIFN-gamma, but not irrelative proteins such as Ag85B, ESAT-6-CFP-10 and GM-CSF, suggesting that the four hybridoma cell lines were rBovIFN-gamma specific monoclonal antibodies. A sandwich ELISA was established by using A10 secreted monoclonal antibody and rabbit polyclonal antibodies against BovIFN-gamma, HRP labeled goat anti-rabbit IgG. The results indicated that the sensitivity was 2ng/mL. This sandwich ELISA to detect BovIFN-gamma paved the way to develop a sensitive method for specific infection detection such as bovine tuberculosis diagnosis.
Animals ; Animals, Newborn ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; immunology ; Blotting, Western ; Cattle ; Cells, Cultured ; DNA, Complementary ; genetics ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Hybridomas ; Interferon-gamma ; genetics ; immunology ; metabolism ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Rabbits ; Recombinant Proteins ; immunology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the value of immunomarkers CXCL13, CD10, bcl-6 in pathologic diagnosis of angioimmunoblastic T-cell lymphoma (AITL).

METHODSOne hundred and fifteen cases of AITL, 30 cases of peripheral T-cell lymphoma, not otherwise specified (PTCL, NOS) and 30 cases of reactive lymph nodes with paracortical hyperplasia (RH) encountered during the period from January, 1990 to January, 2008 were retrieved from the archival files of the Department of Pathology, West China Hospital of Sichuan University, China. The morphologic features were reviewed and compared. Immunohistochemical study was performed by SP method for CXCL13, CD10, bcl-6, CD21, CD3epsilon, CD3, CD45RO, CD20 and Ki-67. TCR-gamma gene rearrangement study was also carried out.

RESULTSRegressed follicles were evident in 7.8% (9/115) of AITL cases, 6.7% (2/30) of PTCL, NOS cases and 83.3% (25/30) of RH cases, respectively. A marked increase of number of arborizing venules was shown in 98.3% (113/115) of AITL cases, 63.3% (19/30) of PTCL, NOS cases and 76.7% (23/30) of RH cases, respectively. In lymph nodes with paracortical hyperplasia, the expression of CXCL13, CD10 and bcl-6 were restricted to the germinal centers. In AITL, 96.5% (111/115) of cases showed CXCL13 expression, in contrast to 26.7% (8/30) of PTCL, NOS. Expression of CD10 and bcl-6 were found in the neoplastic cells in 50.4% (58/115) and 78.3% (90/115) of AITL, and 3.3% (1/30) and 3.3% (1/30) of PTCL, NOS, respectively. Irregular meshworks of CD21-positive follicular dendritic cells were found in all the AITL cases. Clonal TCR-gamma rearrangement was detected in 83% (83/100) of the AITL cases.

CONCLUSIONSAITL is a type of lymphoma originated from the follicular helper T cells. Detailed morphologic assessment and use of immunohistochemical markers are essential for accurate diagnosis.

Adult ; Aged ; Aged, 80 and over ; Chemokine CXCL13 ; metabolism ; Diagnosis, Differential ; Female ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Humans ; Immunoblastic Lymphadenopathy ; metabolism ; pathology ; Lymph Nodes ; metabolism ; pathology ; Lymphoma, T-Cell, Peripheral ; metabolism ; pathology ; Male ; Middle Aged ; Neprilysin ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; Pseudolymphoma ; metabolism ; pathology

OBJECTIVETo investigate the comparability of bcr-abl (P210) transcript levels detected in different hospitals.

METHODSTen hospitals in China took part in the four times of sample exchange and comparisons from April, 2010 to August, 2011. The exchange samples were prepared by Peking University People's Hospital. Firstly, the BCR-ABL (P210)(+) cells from a newly diagnosed chronic myeloid leukemia patient were 10-fold serially diluted by BCR-ABL (P210)(-) cells and they covered 4 magnitudes. Then, TRIzol reagents were thoroughly mixed with cells in each tube. Every 12 samples (three samples per magnitude) were sent to the other 9 hospitals. The cell number of each sample was 8×10(6). The detection of bcr-abl transcript levels by real-time quantitative PCR were performed in every hospital according to their own protocols. Conversion factors (CF) were calculated using regression equation.

RESULTSDifferences in bcr-abl transcript levels did exist among results of 10 hospitals in each comparison. In general, the results of the most of hospitals were in line with the dilutions of cells. CF of every hospital fluctuated. Three hospitals had relatively stable CF, and their ranges were 2.8 - 5.2, 1.2 - 2.8 and 2.2 - 6.8, respectively; two hospitals had unstable CF with ranges 0.76 - 7.0 and 2.1 - 18.7; three hospitals couldn't be calculated CF one or two times because of the significant deviation of the results from the actually bcr-abl transcript levels, and their ranges of CF which could be calculated were 1.9 - 19.2, 3.6 - 7.6 and 0.18 - 14.7; One hospital only had two CF (3.3 and 5.0) because of the replacement of an important reagent during the period of comparisons.

CONCLUSIONSComparability of bcr-abl (P210) transcript levels between different hospitals could be achieved through CF which acquired by sample exchange and comparison. The stable and reliable detection system is the premise to acquire correct CF.

Bone Marrow Cells ; China ; Fusion Proteins, bcr-abl ; genetics ; isolation & purification ; Hospitals ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To further explore the genetic characteristics of oidiomycetes mutant strains like bacterial morphology on the basis of the study on morphology and structure of mutated candida.Methods The standard strains of candida albicans were induced by low temperature and under the condition of low temperature and nutrient deficiency.Variation of standard strains of Candida albicans were induced by clinical antifungal drugs such as fluconazole with different concentration gradient.Fungal gene template was prepared by boiling method,sequences of 16SRNA and 18SRNA were amplified using bacteria conservative gene sequence of 16SRNA and fungal conserved gene sequence of 18SRNA,and observed and recorded the results agarose gel electrophoresis.At the same time,the amplified fragment of bacterial conservative gene 16SRNA was sequenced,and the sequence was analyzed by BLAST comparison.Results the 16SRNA sequences of candida variant were amplified positive,while the standard strain of candida albicans did not show the corresponding amplification band.Except 2 strains which showed a faint band,the other variants of the 18SRNA sequences did not amplified the target band,while the standard strains of candida albicans showed a corresponding amplification bands.Suggested that proportion of 18SRNA sequences in the genome of oidiomycetes mutant strains like bacterial morphology was not much even lack.The 16SRNA fragments amplified of oidiomycetes mutant strains like bacterial morphology did determination of DNA sequence after purification.BLAST comparison analysis,it was found that sequence of oidiomycetes mutant strains like bacterial morphology had higher similarity with bacterial sequences in the database.Conclusion Oidiomycetes mutant strains like bacterial morphology contained bacterial and a small amount of fungus conservative gene.Oidiomycetes mutant strains like bacterial morphology with original nuclear biological character are ones from eukaryotes.This study is great significance in biological evolution,especially in the evolution of prokaryotic cells and eukaryotic cells.

OBJECTIVE: The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipid-associated membrane proteins (LAMPs) in the host innate immune system, specifically their effect on Toll-like receptors (TLRs). METHODS: LAMPs were derived from U. urealyticum strains, and human amniotic epithelial cells (HAECs) were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay (ELISA) and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. RESULTS: LAMPs induced HAECs to produce inflammatory cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor (anti-hTLR2-IgA). CONCLUSIONS LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.
Amnion/cytology* ; Amniotic Fluid/cytology* ; Cytokines/metabolism* ; Dose-Response Relationship, Drug ; Epithelial Cells/metabolism* ; Female ; Humans ; Inflammation ; Interleukin-6/metabolism* ; Interleukin-8/metabolism* ; Lipids/chemistry* ; Lipopolysaccharides/metabolism* ; Membrane Proteins/metabolism* ; Placenta/metabolism* ; Pregnancy ; Toll-Like Receptor 2/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Up-Regulation ; Ureaplasma urealyticum/metabolism*

BackgroundSerum cryptococcal antigen (CrAg) test is the most used noninvasive method to detect cryptococcal infection. However, false-negative CrAg test is not uncommon in clinical practice. Then, the aim of this study was to investigate the factors associated with false-negative CrAg test among non-human immunodeficiency virus (HIV) adult patients with pulmonary cryptococcosis and its clinical features.

MethodsOne hundred and fourteen non-HIV adult patients with pulmonary cryptococcosis, proven by biopsy, were retrospectively reviewed. Finally, 85 patients were enrolled; 56 were CrAg positive (CrAg+ group) and 29 were negative (CrAg- group). It was a cross-sectional study. Then, baseline characteristics, underlying diseases, clinical symptoms, laboratory findings, and chest radiological findings were reviewed and analyzed. Chi-square test was used to analyze categorical variable. Odds ratio (OR) was used to measure correlation. Student's t- test was obtained to analyze continuous variable.

ResultsNo difference in baseline characteristics, underlying diseases, clinical symptoms, and laboratory findings were found between two groups (P > 0.05 in all). Nevertheless, diffuse extent lesion was 82.1% in CrAg+ group and 10.3% in CrAg- group (χ = 40.34, P < 0.001; OR = 39.87).

ConclusionsAmong patients with limited pulmonary involvement, a negative serum CrAg does not preclude the diagnosis of pulmonary cryptococcosis. However, among patients with extensive pulmonary involvement, serum CrAg is a useful diagnostic tool for pulmonary cryptococcosis. Furthermore, we also noticed that the untypical and mild presentations with extensive pulmonary lesion might be the features of pulmonary cryptococcosis, which needs further investigation.

Adolescent ; Adult ; Cross-Sectional Studies ; Cryptococcosis ; immunology ; pathology ; Humans ; Lung Diseases ; immunology ; pathology ; Male ; Retrospective Studies