Objective To study the clinical characteristics and treatment of primary nasal B-cell lymphoma (PNBCL). Methods A retrospective analysis was performed based on the clinical records of 18 PNBCL cases who were treated from January 2009 to June 2015. The clinical manifestations, imaging features, diagnosis approaches and treatment of them were analyzed. Results The main symptoms were nasal obstruction and rhinorrhea. Of all patients, 15 cases were in Ann Arbor stageⅠE-ⅡE, and 3 cases were in Ann Arbor stageⅢE-Ⅳ. The median age was 51 years (12-76 years). The ratio of men to women was 11:7. Only 1 patient had B symptoms. Elevated LDH levels were observed in 4 patients. 13 patients were diffuse large B-cell lymphoma(DLBCL), 3 patients were mantle cell lymphoma, and 2 patients were Burkitt lymphoma. CT examination showed the abnormal nasal soft tissue shadow, with unilateral location and light to moderate enhancement. 14 patients received combination chemotherapy only, and 3 patients received chemotherapy and radiotherapy. Total effective rate was 82.3 % (14/17). At the time of last follow-up, 5 patients died, and the 3-year OS rate was 54.5%(6/11). Conclusions Most PNBCL patients are in Ann Arbor stageⅠE-ⅡE and B symptoms are rare, and the most common pathological types is DLBCL. The treatment for PNBCL is chemotherapy, radiotherapy can assist, but the prognosis is poor, and innovative chemotherapy regimens are necessary.

OBJECTIVETo investigate the feasibility of promoting nerve regeneration by using microcapsulated NGF-expressing cells transplantation.

METHODSThe plasmid pcDNA3. 1 + /NGF, containing rat NGF gene, was transfected into the NIH 3T3 cell by using FuGENE6 transfection reagent. The genetically modified cells expressing NGF gene were enclosed within the alginate-polylysine-alginate (APA) microcapsules and then cultured in vitro. The growth and NGF secretion of the cells were measured periodically. At the same time, these microcapsulated NGF-expressing cells were transplanted into the injured sciatic nerve. The regeneration and functional recovery of the nerve were evaluated in 4 weeks, 8 weeks and 12 weeks after the operation.

RESULTSThe microcapsulated cells had survived and secreted the NGF in three months in vitro. In the group with microcapsulated NGF-expressing cells, the number of the regenerated axons was in large and the nerve fibers were arranged regularly. Compared to other groups, there was less scar , edema and monocytes found at the stoma in the goup. The moter nerve conductive velocity, nerve muscle-action potential and SFI were improved.

CONCLUSIONSThe microcapsulated NGF-expressing cells could significantly enhance the nerve regeneration and reduce inflammatory response of xenograft.

Animals ; Cell Transplantation ; Female ; Male ; Mice ; NIH 3T3 Cells ; Nerve Growth Factor ; biosynthesis ; genetics ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo analyze and optimize the gene rearrangement primers of different frame regions (FR) of immunoglobulin heavy chain (IgH) genes by bioinformatic methods and explore the application of these primers in the detection of paraffin-embedded lymphoma tissues.

METHODSThree pairs of primers from IgH FR1, FR2 and FR3 regions (P1c, P2A and P31, respectively) were selected as the B cell gene rearrangement primers after comparison of the gene fragments in 44 IgH variable and 6 joining regions. Using one pair of T cell receptor (TCR) gamma primer as the T cell gene rearrangement primer, 101 histopathologically confirmed lymphoproliferative samples including 80 B cell lymphomas, 14 T cell lymphomas, and 7 reactive proliferative lymph nodes were examined by PCR for gene arrangement. The DNAs from DG75 and Jurkat cell lines were used as the positive controls for B and T cell lymphoma, respectively, with those from reactive proliferative lymph nodes as the negative control.

RESULTSThe positivity rates of IgH primers (P1c, P2A and P31) in the 80 B cell lymphomas were 37.5% (30/80), 52.5% (42/80) and 70.0% (56/80), respectively, and only one of the 14 T cell lymphoma cases was positive for the primers, suggesting significant differences in the detection rates of B cell lymphomas by the 3 primers. The detection rate was increased to 83.9% by combining the results by P31 and P2A primers. No positivity was found in the proliferative reaction tissues.

CONCLUSIONPrimers from IgH FR3 region genes are more sensitive than that from the FR1 and FR2 regions in the detection of gene rearrangement in paraffin-embedded lymphoma tissues. The detection rates can be increased by combining the results with the primers for IgH FR3 with that of FR2.

DNA Primers ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; genetics ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Lymphoma, B-Cell ; diagnosis ; genetics ; pathology ; Lymphoma, Non-Hodgkin ; diagnosis ; genetics ; pathology ; Lymphoma, T-Cell ; diagnosis ; genetics ; pathology ; Male ; Paraffin Embedding

192 samples of Masked Palm Civet (Paguma Larvata) from Guangdong Province were inoculated in Vero-E6 cells. One sample which came from masked palm Civet didn't cause cytopathic effects (CPE) until fourth-passage on Vero-E6 cells. Infected cells emerged granulating, shrinking, rounding and falling off. After three times freeze-thaw, cells and culture medium were harvested for electron microscopy. Virus particles were nonenveloped, double capsid and icosahedral symmetry. This virus was designated Masked Palm Civet/China/2004 (MPC/04). Hemagglutination test indicated that the virus could agglutinate healthy human type O red cells, but not the red cells of SPF chicken, experimental common bovine, rat and guinea pig. This virus was tolerant to chloroform treatment, pH 3.0 and water bath 50 degrees C 1 h. 1 M MgCl2 treatment could enhance resistance of virus to heat and increase infectivity. In order to classify the strain on the molecular level, specific primers according to mammalian reovirus were used for Reverse Transcription Polymerase Chain Reaction (RT-PCR). Appropriate specific products were amplified by RT-PCR. NCBI BLAST analysis indicated that this segment shared the highest identity to mammalian reovirus serotype 1 (T1L) virus. So we can deduce this virus is a member of the Reoviridae.
Animals ; Base Sequence ; Cats ; virology ; Cattle ; Humans ; Molecular Sequence Data ; Phylogeny ; Reoviridae ; classification ; isolation & purification

Membrane (M) protein genes of 20 infectious bronchitis virus (IBV) strains isolated in China between 1995 and 2004 were sequenced and analyzed. The M genes of twenty isolates were composed of 672 to 681 nucleotides, encoding polypeptides of 223 to 226 amino acid residues. Variations of the deduced amino acids of M gene mainly occurred at positions 2 to 17 and 221 to 233, comparing with that of the IBV strain LX4. There were deletions or insertions in the M gene of Chinese isolates at amino acid position 2 to 6, leading to the loss or gain of a glycosylation site. Phylogenetic tree based on amino acid sequences of M genes from 20 Chinese isolates and 34 reference strains showed that they were classified into five distinct clusters. Most of the Chinese IBV strains were included in clusters II and IV, forming distinct groups. The isolates in cluster II showed a close evolutionary relationship with Taiwan isolates. Furthermore, recombination especially the recombination between field isolates and vaccine strains had been observed while comparing the phylogeny of M genes with those of S1 and N genes.
Amino Acid Sequence ; China ; Genetic Variation ; Infectious bronchitis virus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Sequence Homology, Amino Acid ; Viral Matrix Proteins ; genetics

OBJECTIVETo construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells.

METHODSThe full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry.

RESULTSA gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells.

CONCLUSIONA eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.

12E7 Antigen ; Antigens, CD ; biosynthesis ; genetics ; Base Sequence ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Cell Line, Tumor ; Cloning, Molecular ; Genetic Vectors ; genetics ; Hodgkin Disease ; metabolism ; pathology ; Humans ; Jurkat Cells ; Molecular Sequence Data ; Transfection

OBJECTIVETo study the expression pattern of cd99l2 gene during zebrafish development, the RNA probes for whole-mount in situ hybridization were prepared in this study.

METHODSThe cd99l2 fragment obtained by RT-PCR was cloned into pGM-T Easy, then the plasmids were linearized with the restriction enzymes SacII or SalI. Using Sp6 or T(7) RNA polymerase, the digoxingenin-labeled antisense and sense probes were synthesized and confirmed by whole-mount in situ hybridization.

RESULTSThe plasmid cd99l2/pGM-T was constructed. cd99l2 gene expression pattern during embryogenesis of zebrafish was examined using the antisense probe, and intense expression was detected in the central nervous system during zebrafish development.

CONCLUSIONThe antisense probe can be used for study of the spatial and temporal distribution of cd99l2 during zebrafish development using the sense probe as control.

Animals ; Central Nervous System ; embryology ; Cloning, Molecular ; Digoxigenin ; chemistry ; Gene Expression Regulation, Developmental ; Oligonucleotide Probes ; RNA Probes ; Uridine Triphosphate ; chemistry ; Zebrafish ; embryology ; genetics ; Zebrafish Proteins ; genetics

The entire S1 protein gene of five infectious bronchitis (IB) vaccine strains (JAAS, IBN, Jilin, J9, H120) used in China were compared with that of the IB field isolate CK/CH/LDL/97 I present in China. The nucleotide and deduced amino acid similarities between the five IB vaccine strains and the field strain, CK/CH/LDL/97 I, were not more than 76.4% and 78.7%, respectively. Phylogenetic analysis based on the S1 gene showed that the vaccine strains and the field strain belonged to different clusters and had larger evolutionary distances, indicating that they were of different genotypes. The five vaccine strains were used for protection test against challenge of the field isolate CK/CH/LDL/97 I. The chickens inoculated with five vaccine strains showed morbidity as high as 30%-100% after challenged with the CK/CH/ LDL/97 I strain. The organ samples at 5 days post challenge showed that the viral detection rates were 50%-90% and 10%-30% for trachea and kidney, respectively. The live attenuated vaccines only provided partial protection to the vaccinated chickens against heterologous IBV infection, CK/CH/LDL/97 I.
Animals ; Antibodies, Viral ; blood ; Chickens ; virology ; Coronavirus Infections ; prevention & control ; veterinary ; Infectious bronchitis virus ; classification ; genetics ; immunology ; isolation & purification ; Membrane Glycoproteins ; genetics ; Phylogeny ; Poultry Diseases ; prevention & control ; Spike Glycoprotein, Coronavirus ; Vaccines, Attenuated ; immunology ; Viral Envelope Proteins ; genetics ; Viral Vaccines ; immunology