The Japanese encephalitis virus (JEV), a leading cause of encephalitis, exists as quasispecies in clinical isolates. Using a limiting dilution method combined with immunohistochemistry to detect viral antigens, 10 biological clones were isolated and purified from a clinical JEV isolate (CNS138/9) derived from an autopsy brain. These biological clones were tested for neurovirulence in SK-N-MC and NIE-115 neuronal cells, and a 2-week-old, footpad-infected, JE mouse model. Nine clones were found to be neurovirulent; one clone neuroattenuated. Although further studies are needed to determine genotypic differences, if any, in these clones, the limiting dilution purification and neurovirulence testing methods described herein should be useful for phenotypic studies of quasispecies of neurotropic viruses in general, and JEV and other flaviviruses in particular.

In this study, we evaluated the difference ot biological characteristics in the MERS-CoV infected mice model in prior to transduction with different dosage of human DPP4. Firstly, we transduced different dosage of DPP4 (high or low) into mice, and then challenged them with MERS-CoV in order to establish the model. After establishment of mice model, we observed the clinical signs of disease, virus replication, immunopathogenesis and antibody response. The results indicated that the infected mice showed typical pneumonia, virus replication, histological lesions, and neutralizing antibody production. Moreover, the high dosage group was superior to the low dosage group. Fourteen days after infection, the specific antibody to virus structural protein and neutralizing antibody were analyzed, the high dosage group induced higher level antibody. In summary, the MERS-CoV infected mice model were established prior transduction with DPP4, and the level of DPP4 influenced the clinical signs of disease, virus replication and antibody response in this model.
Animals ; Coronavirus Infections ; enzymology ; genetics ; pathology ; virology ; Dipeptidyl Peptidase 4 ; genetics ; metabolism ; Disease Models, Animal ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Middle East Respiratory Syndrome Coronavirus ; genetics ; physiology

OBJECTIVETo study the characteristics of allergic reactions to common aeroallergens in young children with wheezing or allergic diseases by examining the results of skin prick test in children under 5 years old.

METHODSA total of 196 children under 5 years old, from a district of Changsha City sampled between September 1 to December 31, 2010, were assigned into two groups according to the presence of wheezing or allergic diseases: allergen screening (n=102) and control (n=94). Skin prick tests were performed on both groups.

RESULTSThe positive rate of skin prick test in the allergen screening group was 61.8% (63/102), and this was significantly higher than in the control group (9.6%, 9/94; P<0.05). In the allergen screening group, the positive rate of skin prick test in children with both recurrent wheezing and allergic rhinitis was significantly higher than in children with wheezing alone (P<0.05). The frequency of wheezing was positively correlated with a positive skin prick test (r=0.91; P<0.05). The positive rate of skin prick test for mites was significantly higher than for other aeroallergens (24.2% vs 3.5%; P<0.05) in the allergen screening group. Skin prick testing of the children for dermatophagoides farinae showed a higher positive rate than for dermatophagoides pteronyssinus (50.0% vs 14.7%; P<0.05).

CONCLUSIONSWheezing in early childhood may be associated with the occurrence of asthma. Skin prick testing contributes to the diagnosis of allergic diseases and assessment of allergic reactions to aeroallergens in children with wheezing.

Asthma ; etiology ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Sounds ; diagnosis ; immunology ; Rhinitis, Allergic, Perennial ; diagnosis ; immunology ; Rhinitis, Allergic, Seasonal ; diagnosis ; immunology ; Skin Tests

OBJECTIVETo study the effects of suplatast tosilate (IPD) on the airway inflammation and expression of interleukin-5 in asthmatic rats.

METHODSFifty adult male Sprague-Dawley rats (4-week- old) were randomly assigned to five groups: placebo control, untreated asthma, budesonide(BUD)-treated asthma , early or late IPD intervention group (n=10 rats each). Asthmatic mode was prepared by ovalbumin sensitizion and challenge. Inflammatory cells and the percentage of EOS were detected in bronchoalveolar lavage fluid (BALF). The lung tissues were removed to detect the lung histomorphology. Gene expression of IL-5 was measured by reverse transcription-polymerase chain reaction (RT-PCR). Levels of interleukin 5 (IL-5) in BALF were measured using ELISA.

RESULTSThe inflammatory cells and the percentage of EOS in BALF, IL-5 levels in BALF and IL-5 mRNA expression in the lung tissues were obviously higher in the untreated asthma group than the control group (P<0.05), while the parameters in the IPD or BUD-treated asthma groups were significantly lower than the untreated asthma group (P<0.05).

CONCLUSIONSIPD treatment can alleviate airway inflammation in asthmatic rats, possibly through inhibiting IL-5 mRNA transcripts.

Animals ; Arylsulfonates ; therapeutic use ; Asthma ; drug therapy ; immunology ; pathology ; Eosinophils ; drug effects ; Interleukin-5 ; analysis ; antagonists & inhibitors ; genetics ; Lung ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Sulfonium Compounds ; therapeutic use

OBJECTIVETo observe the patterns of benign and malignant breast lesions using real-time gray-scale contrast-enhanced ultrasound and assess its value in the differential diagnosis of begign and malignant breast tumors.

METHODSTotally 116 breast lesions (benignity n = 63; malignancy n = 53) underwent real-time gray-scale contrast-enhanced ultrasound. The patterns of their enhancement were assessed from 6 aspects: degree of enhancement, process of enhancement, completeness of enhancement, homogeneity of enhancement, boundary of the enhanced lesions, and exhistance of radial enhancement around the lesions. The results were compared with the pathologic findings.

RESULTSContrast-enhanced sonographic patterns were significantly different between benign and malignant breast lesions. Most malignant lesions were non-centripetally, incompletely, and inhomogeneously enhanced. After having been injected with the microbubble contrast medium, the boundary of the lesions became unclear, and the radial enhancement around lesion were mainly seen in the malignant lesions.

CONCLUSIONThe patterns of real-time gray-scale contrast-enhanced ultrasound are remarkably different between malignant and benign breast lesions, showing promising values for its clinical application.

Breast Neoplasms ; diagnostic imaging ; Contrast Media ; administration & dosage ; Diagnosis, Differential ; Female ; Humans ; Microbubbles ; Ultrasonography, Mammary

OBJECTIVETo observe the performance of liver metastases with contrast-enhanced ultrasound (CEUS) and assess its clinical application.

METHODSTwenty-one patients with 21 untreated liver metastases underwent CEUS with low mechanical index imaging. The characteristic appearances of CEUS in different vascular phases were observed.

RESULTSOf 21 metastases, 19 (90.5%) were identified as fast-in and fast-out enhancement pattern. In the arterial phase, all the 21 lesions showed enhancement but with varied appearances: 12 (57.1%) showed early diffuse enhancement, 8 (38.1%) showed ring-like enhancement, and the remaining one lesion of large size showed slowly enhomogenous minor enhancement. In the late phase, sharp defects were found in 20 lesions (95.2%), and more lesions were detected in 3 patients (14.3%).

CONCLUSIONCEUS can show the characteristic appearance of liver metastases, and can be used for the screening and diagnosis of liver cancers.

Contrast Media ; Humans ; Liver Neoplasms ; diagnostic imaging ; secondary ; Ultrasonography

T-cell internal antigen-1 (TIA-1) has roles in regulating alternative pre-mRNA splicing, mRNA translation, and stress granule (SG) formation in human cells. As an evolutionarily conserved response to environmental stress, SGs have been reported in various species. However, SG formation in chicken cells and the role of chicken TIA-1 (cTIA-1) in SG assembly has not been elucidated. In the present study, we cloned cTIA-1 and showed that it facilitates the assembly of canonical SGs in both human and chicken cells. Overexpression of the chicken prion-related domain (cPRD) of cTIA-1 that bore an N-terminal green fluorescent protein (GFP) tag (pntGFP-cPRD) or Flag tag (pFlag-cPRD) induced the production of typical SGs. However, C-terminal GFP-tagged cPRD induced notably large cytoplasmic granules that were devoid of endogenous G3BP1 and remained stable when exposed to cycloheximide, indicating that these were not typical SGs, and that the pntGFP tag influences cPRD localization. Finally, endogenous cTIA-1 was recruited to SGs in chicken cells and tissues under environmental stress. Taken together, our study provide evidence that cTIA-1 has a role in canonical SG formation in chicken cells and tissues. Our results also indicate that cPRD is necessary for SG aggregation.
Chickens ; Clone Cells ; Cycloheximide ; Cytoplasmic Granules ; Humans ; Protein Biosynthesis ; RNA Precursors ; RNA-Binding Proteins ; T-Lymphocytes

Humans ; Carcinoma, Hepatocellular/epidemiology* ; Liver Neoplasms/epidemiology* ; Prothrombin ; Biomarkers

OBJECTIVETo investigate the prognostic value of morphology and Hans classification in diffuse large B cell lymphoma（DLBCL）.

METHODSClinical data of 249 patients diagnosed with DLBCL in our hospital and Hangzhou Xixi hospital during Jan 2006 to Dec 2016 were analyzed retrospectively. These patients were classified into 3 groups: immunoblastic variant（IB） group, centroblastic variant（CB） group and others group according to the cell morphology. And DLBCL was also divided into GCB（germinal center B-cell-like）or non-GCB（non-germinal center B-cell-like） group by analyzing the expression of CD10, BCL6 and MUM1 (GCB: CD10 ,BCL6,MUM1/CD10,BCL6,MUM1；non-GCB：CD10,BCL6,MUM1/CD10,BCL6,MUM1).

RESULTSThe univariate analysis displayed that the age，LDH level，IPI，IB，non-GCB，B-symptoms and rituximab all could influence the OS and EFS, the CR rate of CB subtype patients was significantly higher than that of the patients with IB subtype （68.3% vs 38.9%)(P=0.02）. IB subtype was the in dependent prognostic factor for both EFS and OS in the whole study. In multivariate analysis, IPI and IB were the independent prognostic factors for OS and EFS. IB subtype was also an independent prognostic factor in EFS and OS with or without rituximab. The expression of BCL2 and BCL6 was related with prognosis in R-CHOP, but not in CHOP treated patients. Other markers (CD5, CD10, IRF4/MUM1, HLA-DR and Ki-67 proliferation index) were not of the significant prognostic value for DLBCL. When accepted rituximab, the GCB and non-GCB were not different significantly for prognosis. However, the non-GCB group showed a poor prognosis without using rituximab （EFS P=0.020；OS P=0.020）. Multivariate Cox models showed that OS and EFS were not significantly different between GCB and non-GCB group, however, the IB subtype had a very significantly poor prognosis in OS and EFS (P=0.001, P=0.002). When the analysis was restricted to DLBCL with CB morphology only, no prognostic value was observed in Hans classification.

CONCLUSIONThe subtype of immunoblast is a major risk factor in patients treated with CHOP or R-CHOP. There is a significant association between the Hans classification and the morphologic subclassification. Results of this study have supplemented the data for the prognostic factor of DLBCL and demonstrated that the cytomorphologic diagnosis can be reproducible.

Antineoplastic Combined Chemotherapy Protocols ; Cyclophosphamide ; Doxorubicin ; Humans ; Immunohistochemistry ; Lymphoma, Large B-Cell, Diffuse ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Rituximab