Objective To construct the eukaryotic expression vector of linker for activation of T cells(LAT) gene in mouse. MethodsLAT cDNA of mouse mononuclear cells was amplified with polymerase chain reaction.The fragment was inserted into the eukaryotic expression vector of pCMV-HA plasmid.The recombinant plasmid was verified by DNA sequencing and used to transfect lymphocytes of the asthmatic model. Results The length of amplified fragment was 729 bp.The sequence of LAT gene was confirmed by Genbank.The LAT protein could be detected in the lymphocytes of asthmatic model.Conclusion The recombinant eukaryotic expression vector pCMV-HA-LAT can be successfully constructed.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast ; pathology ; Breast Neoplasms ; drug therapy ; pathology ; Cyclophosphamide ; therapeutic use ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Lymphoma, B-Cell ; drug therapy ; pathology ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; pathology ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Sarcoma ; pathology ; Spleen ; pathology ; Splenic Neoplasms ; drug therapy ; pathology ; Vincristine ; therapeutic use

Objective To establish a mouse model of H.pylori infection, and to evaluate the chronic pathological changes in the gastric mucosa associated with H.pylori infection.Methods 34 male 5~6-week old SPF C57BL/6 mice were used in this study.The mice were intragastrically administrated with a suspension of H.pylori SS1 strain.Two weeks after infection, rapid urease test and PCR were performed to confirm the H.pylori infection.Successfully infected mice were randomly divided into 3 groups including the control group, 6-week and 12-week infected groups.Samples of gastric mucosa were taken for pathological analysis using HE and borax methylene blue staining.The contents of myeloperoxidase (MPO), superoxide dismutase (SOD), malondialdehyde (MDA) and catalase (CAT) in the gastric tissues were detected by biochemistry, and the expression levels of COX-2, iNOS, TNF-α and IL-1β were examined by RT-qPCR.Results Compared with the control group, H.pylori colonization was observed in the gastric mucosa of the 6-week and 12-week infected mice, with chronic inflammatory cell infiltration, glandular atrophy and intestinal metaplasia to varying extents.The contents of CAT and SOD were significantly decreased, while the levels of MPO and malonaldehyde MDA, and the expression levels of COX-2,iNOS,TNF-α and IL-1β were significantly increased (P<0.05 or P<0.01).Conclusions Intragastric administration with H.pylori in C57BL/6 mice can be successfully used to generate the bacterial colonization, leading to chronic inflammatory cell infiltration, enhanced oxidative stress, and up-regulated expression of proinflammatory genes in the gastric glandular tissues at 6 and 12 weeks after inoculation.However, the inflammatory changes are more extensive in the mice at 12 weeks after infection, with glandular atrophy and intestinal metaplasia.

Objective:To compare two flexible embryo catheters and determine whether clinical out-come differs in the in vitro fertilization-embryo transfer (IVF-ET)cycles.Methods:This prospective control study was conducted by one doctor between July 2012 and November 2013.In the study,2 064 patients undergoing fresh embryo transfer by using IVF-ET/intracytoplasmic sperm injection (ICSI)-ET in Reproductive Medical Center of Peking University Third Hospital were recruited.The subjects were di-vided into two groups.Cook Sydney IVF embryo transfer catheters (product model:K-JETS-7019-SIVF) were used for embryo transfer in group 1 (n =949),and FrydmanCCD catheters (product model:131230301)were used in group 2 (n =1 115).Pregnancy outcomes were compared between these two groups.Results:There was no significant difference in age,diagnosis for infertility and stimulation proto-col used between the two groups.In addition,there was no difference in the number of oocytes collected and in the number and score of embryos transferred.The significantly higher implantation rate,clinical pregnancy rate,and live birth rate (34.40% vs.26.92%,51.21% vs.41.52%,42.57% vs. 33.09%,P <0.05)were observed in group 1 compared with group 2.The abortion rate was not signifi-cantly different between the two groups (11.93% vs.15.98%,P >0.05).The proportion of difficult transfer was higher in group 1 than that in group 2 (5.27% vs.3.41%,P <0.05 ).There was no difference in the clinical pregnancy rate and live birth rate between the two difficult transfer cycles.Con-clusion:The type of embryo transfer catheter affects the clinical outcome in IVF.Good clinical outcome can be obtained by using Cook Sydney IVF catheter,which is worthy of clinical promotion.

Humans ; Siblings ; Forensic Medicine ; Forensic Genetics

BACKGROUNDAllergic asthma is associated with airway inflammation and hyperresponsiveness caused by dysregulated production of cytokines secreted by allergen-specific helper T-type 2 (Th2) cells. The linker for activation of T cells (LAT) is a membrane-associated adaptor protein, which has been shown to take part in regulating T cell receptor (TCR) signaling and T cell homeostasis. In this study, we established an asthmatic mouse model to examine the changes in LAT levels during allergic airway disease and the effects of LAT transgenic expression on airway inflammation.

METHODST cells from mouse lung tissues were isolated from allergen challenged (ovalbumin (OVA)) and control mice, and the purity of these isolated T cells was examined by fluorescence-activated cell sorter (FACS). Semi-quantitative RT-PCR and Western blotting were used to detect the expression of the LAT gene and LAT protein, respectively. After an intranasally administered mixture of pCMV-HA-LAT plasmid and Lipofectamine 2000, 24 hours before and 72 hours after allergen challenge, the BALF cell count and the differential cytologies were studied. In addition, IL-4 and IFN-γ levels in the BALF were determined by ELISA, and pathological changes in lung tissues were observed.

RESULTSLAT protein and mRNA expression were decreased in lung T cells in a mouse model of allergen-induced airway disease. After intranasal administration of pCMV-HA-LAT, histopathological examination of the lungs showed that intervention with LAT overexpression prevented mice from developing airway inflammation, and the number of total cells, eosinophils, neutrophils, and lymphocytes in the BALF was reduced significantly compared with the OVA sensitized and challenged group. In addition, the Th2 cytokine IL-4 decreased, while the Th1 cytokine IFN-γ increased compared to the OVA sensitized and challenged group or the OVA sensitized group plus pCMV-HA treatment.

CONCLUSIONThis study demonstrates that LAT might effectively diminish Th2 cytokine responses, lung histopathological changes and lung inflammation to allergen challenge in a model of experimentally induced asthma.

Animals ; Asthma ; immunology ; metabolism ; Blotting, Western ; Bronchoalveolar Lavage Fluid ; immunology ; Cells, Cultured ; Cytokines ; metabolism ; Female ; Inflammation ; immunology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocytes ; immunology ; metabolism

BACKGROUNDTp15, Tp17, Tp45, and Tp47 are outer-membrane proteins found in Treponema pallidum, the etiologic agent of syphilis. These proteins are potent antigens and are potential markers for the serological detection of syphilis. The present study analyzed antibodies to these protein antigens (TP-IgM and TP-IgG) in human serum and investigated the expression of these antibodies during different stages of syphilis.

METHODSSerum samples were collected from 69 subjects (male 45, female 24) diagnosed with syphilis and analyzed by Western blotting for the expression of IgM and IgG against the four protein antigens. Expression levels of the target antibodies were compared during the same stage of syphilis as well as between different stages of this disease.

RESULTSIn subjects with primary syphilis, the positive rate of Tp45 IgM was higher than that of other TP-IgM. Tp15 IgM was detected only in subjects with tertiary syphilis. Similarly, the seroprevalence of Tp45 IgG in primary syphilis was higher than for other TP-IgG. No target TP-IgM was detected in subjects with latent syphilis. In subjects with secondary syphilis, the expression level of Tp15 IgG (138.73 ± 20.16) was higher than for other target TP-IgG. In subjects with tertiary syphilis, all target TP-IgG were detected. In subjects with tertiary or latent syphilis, the expression levels of Tp45 IgG (121.33 ± 11.04 and 110.10 ± 40.19, respectively) were higher than those of other target TP-IgG. The expression levels of all Tp-IgM were similar before or after anti-syphilis treatment. In comparison, the expression levels of all TP-IgG decreased compared with the pre-treatment levels, and this decrease was statistically significant (both P < 0.05) for Tp17 IgG and Tp47 IgG.

CONCLUSIONSAfter Treponema pallidum infection, Tp45 IgM appeared first and Tp15 IgM occurred during later stages. The positive rates of all TP-IgG increased with the duration of this disease. Anti-syphilis treatment reduced the expression levels of Tp17 IgG and Tp47 IgG. Larger-scale studies are required to further validate the value of Tp15, Tp17, Tp45, and Tp47 as markers for the early detection of primary and latent syphilis.

Adolescent ; Adult ; Aged ; Antibodies, Bacterial ; blood ; Female ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Male ; Middle Aged ; Syphilis ; diagnosis ; Treponema pallidum ; immunology

OBJECTIVETo describe the clinicopathologic features and immunophenotypes of carcinoid tumorlets in the lung with bronchiectasis, and to study the morphogenesis of these tiny tumors.

METHODSThe histopathologic characteristics of 3 bronchiectasis cases with carcinoid tumorlets, 11 bronchiectasis and 2 normal lungs were studied. Specific markers of the tiny tumors and the number of neuroendocrine cells (NECs) in the airway mucosa were immunohistochemically detected by EnVision method.

RESULTSThe tumorlets in the lungs presented as multi-focal nodules and most were displayed only under microscopy. These cells were arranged in clusters and foci of fascicles which were situated in the surrounding bronchial wall and bronchioles adjacent to bronchiectatic lesion, or in the scar tissues. The tiny tumors were consisted of short fusiform cells and small ovoid cells. Their nuclei were circular, oval or long fusiform and the cells were strongly argyrophilic on Grimelius staining. Intensive positive immunostaining for calcitonin, chromogranin A, NSE and gastrin were detected. Weak positive for CK, EMA, S-100 and focal positive for HC, ACTH, 5-HT were also observed. Proliferative NECs in airway mucosa adjacent to the tiny tumors increased significantly in number, compared with those in the airway mucosa of bronchiectasis without tumorlets and normal lungs (P < 0.001, respectively).

CONCLUSIONSThe clinicopathologic features and immunophenotypes of carcinoid tumorlets resemble carcinoid tumors. They are the early stage of carcinoid development. Their development may be related to the chronic pulmonary damage resulting in hypoxia and stimulating the proliferation of NECs. These pulmonary carcinoid tumorlets can be used as a model to study the tumorigenesis of carcinoid carcinoma of the lung.

Adult ; Aged ; Bronchiectasis ; pathology ; Carcinoid Tumor ; chemistry ; pathology ; Female ; Humans ; Immunohistochemistry ; Lung Neoplasms ; chemistry ; pathology ; Middle Aged ; Morphogenesis ; Neurosecretory Systems ; pathology

OBJECTIVETo explore possible alterations in velopharyngeal function after maxillary advancement.

METHODSTen patients (3 secondary deformity of cleft palate, 7 maxillary retrusion) had maxillary advancement surgery. Pre- and post-operative examinations consisted of lateral cephalometric radiography, nasopharyngoscopy and speech recording.

RESULTSThere was no significant changes on speech or velopharyngeal competence after maxillary advancement. Cleft palate patients appeared slight hypernasality and nasal emission.

CONCLUSIONSGood velopharyngeal function pre-operation is not affected after the surgery of maxillary advancement.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Osteotomy, Le Fort ; Palate ; physiopathology ; surgery ; Pharynx ; physiopathology ; surgery ; Voice Quality ; Young Adult

This study was aimed to investigate the prophylactic effect of Toll like receptor (TLR)5 agonist flagellin on acute graft versus host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and its possible mechanism. The animal model with allo-HSCT aGVHD was established by using purebred mice (male mouse C57BL/6 as donor, female mouse BALB/c as recipient) with complete-unidentical major histocompatibility antigen. The recipient mice were randomly divided into 3 groups: group 1 in which mice were injected with high purity (95%) flagellin before and after allo-HSCT respectively, group 2 in which mice received allo-HSCT without injection of flagellin, group 3 in which mice were radiated alone. The aGVHD features of mice in group 1 and 2 were observed and compared. The results showed that the typical symptoms of aGVHD appeared in transplanted mice. The death peak of mice in group 2 appeared at day 4-5 after transplantation. The aGVHD symptoms were obviously alleviated and the mean survival time was prolonged significantly in mice group 1 as compared with mice in group 2 (P < 0.05). The comparison of WBC count in peripheral blood of mice in 3 groups before transplantation showed no significant difference (P > 0.05), while WBC count of mice in group 1 and 2 showed the significant difference at days 14 and 21 after transplantation (P < 0.05). The pathological appearances of aGVHD in mice of group 1 were obviously reduced as compared with mice in group 2. The flow cytometric detection of Treg cell/CD4(+) T cell levels at different time before and after transplantation demonstrated that the Treg cell level in mice of group 1 at weeks 2-4 after transplantation significantly increased as compared with mice in group 2 (P < 0.05). It is concluded that flagellin can effectively prevent the aGVHD occurrence after allo-HSCT, reduce the symptoms and pathological changes of aGVHD, obviously prolong mean survival time of mice in group 1. The mechanism of flagellin effect may be associated to increase of Treg cell level in mice after allo-HSCT.
Animals ; Female ; Flagellin ; therapeutic use ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes, Regulatory ; Toll-Like Receptor 6 ; agonists ; Transplantation, Homologous