Background Orbital diffuse large B cell lymphoma(DLBCL) is a kind of malignant lymphoma with higher morbidity,but the systematic study is difficult for the fewer cases and the lack of orbital DLBCL cell line.Objective This study was to compare the differences or similarities of histopathological characteristics between orbital DLBCL and systemic DLBCL,and to discuss whether systemic DLBCL cell line is available to in vitro research of orbit DLBCL.Methods The histopathological specimens were collected from 3 orbital DLBCL patients and 15 systemic DLBCL patients in Tianjin Eye Hospital and Tianjin Tumor Hospital from 2004 to 2009.The prognosis of the patients was followed-up,and the survival curve was drew.The histopathological examination of the specimens was performed using hematoxylin and eosin staining,and the expressions of CD20,CD79α,CD45RO,CD10,BCL-6,mum-1,Ki-67 and surviving in the specimens were detected by immunochemistry.The results mentioned above were analyzed and compared between orbital DLBCL and systemic DLBCL.Results The histopathological examination showed that the centroblastic type was the primary form in both orbital DLBCL and systemic DLBCL,according with the diagnosis of DLBCL.The differences in the positive expression rate of CD20,CD79α,CD45RO,Ki-67,survivin were not statistically significant between the two types of DLBCL (P =0.167,0.442,1.000,1.000,0.442).Immunochemistry revealed that 3 orbital DLBCL patients were germinal center B-cell-like (GCB),and 2 of the patients showed the positive expression for BCL-6 and mum-1 as well as absent expression for CD10,but the other patient presented the absent expression for BCL-6,mum-1 and CD10.In 15 systemic DLBCL patients,7 were GCB type,with the positive expression for CD10 in all the 7 patients and absent expression for BCL-6 in 6 patients.In addition,in 8 of non-GCB type,3 appeared to be absent expressed for CD10 and BCL-6,and 5 were positive expressed for BCL-6 and mum-1 and absent expressed for CD10.No significant difference was found in the survival duration between the orbital DLBCL and systemic DLBCL (P =0.067).Conclusions There exists no clinically significant difference in the pathological features,the expression of tumor cell markers and prognosis in both orbital and systemic DLBCL.These two DLBCLs appear to be similar in epidemiology and clinical staging,inferring that systemic DLBCL cell line is available in the in vitro study of orbital DLBCL.

Background Recently, the morbidity of orbital lymphoma increased gradually, and we have made deeper research in pathology,therapy and pathogenesis of the disease.There were few reports of mice model of orbital lymphoma up to now for its lower morbidity and culture difficulty.Objective This study was to establish a mouse model of orbital diffuse large B cell lymphoma (DLBCL) by injection of systemic DLBCL cell line pfeiffer.Methods Ten SPF BALB/c mice and 5 nod-SCID mice were radiated firstly using 137Cs,with the absorption dose 3.5 Gy in the BALB/c mice and 2.6 Gy in the nod-SCID mice,and then pfeiffer cells were intraobitally injected in 4eyes of BALB/c mice (orbital injection group) and suncutaneously injectied in 4 eyes of BALB/c mice and 4 eyes of nod-SCID mice (subcutaneous injection group) at the concentration of 1.5×l08/ml.The developing status of tumors were examined once per day and the growth curve was drawn.The tumors and nearby lymph nodes were obtained 54 days after injection for the preparation of 4 μm thickness of serial sections.Hemotoxylin-eosin staining was used to examine the histopathology of the tumors, and immunochemistry was employed to detect the expressions of CD20,CD79α and CD45RO proteins.The tumors were typed based on the expressions of CD10, BCL-6 and mum-1 in the specimens,and the expressions of Ki-67 and survivin were assyed to assess the prognosis of the tumor.All the results were compared with 3 diagnosed human orbital DLBCL sections.The use and care of the mice complied with Chinese Administration Rule of Laboratory Animal.Results The tumor formation rates were 100％ in both the orbital injection group and subcutaneous injection group, and the tumors grew much faster in nod-SCID mice than BALB/c mice.Infiltration of tumor cells in lymph nodes were found in the subcutaneous injection group rather than the orbital injection group.The pathological features were accordant among the orbital injection group, subcutaneous injection group and human orbital DLBCL sections.The number of ＜50％ and ≥50％ CD20+ specimens was 3 and 5,CD79αwas 2 and 6,CD45RO was 8 and 0 in the BALB/c mice;while that in the nod-SCID mice was 1 and 3 in CD20,0 and 4 in CD79α,4 and 0 in CD45RO;the number of human orbital DLBCL specimens was 1 and 2 in CD20,1 and 2 in CD79α,2 and 1 in CD45RO,without significant differences among them (all at P=1.00).No significant differences were seen in Ki-67+ number and survivin+ number among the BALB/c mice, nod-SCID mice and human orbital DLBCL specimens (all at P=1.00).The detection of CD10,BCL-6 and mum-1 expressions indicated that the tumors of BALB/c mice,nod-SCID mice and human orbital DLBCL specimens all were the non-germinal center B cell-like types.Conclusions The orbital and subcutaneous DLBCL mouse models are successfully established by injection of pfeiffer cell line.There are the same findings and features in biological behavior, pathology and immunohistochemistry in orbital,subcutaneous models with human orbitl DLBCL.Nod-SCID mice appear to be more suitable for the growth of lymphoma cells.

Objective To investigate the effects of staphylococcus aureus (S.aureus) DNA on anti-tumor in vivo and its mechanism. Methods The chromatic DNA purified from S.aureus was used in the experiments of antitumor in vivo, and immunological indices were measured. Results S.aureus DNA could inhibit the tumor growth by peritoneal injection, increase the serum IFN-? level and regulate the serum TNF-? level in the nude mice carrying transplant tumor. Conclusion S aureus DNA has a characteristic of anti-tumor immunity, and can be used in immunotherapeutic treatment of tumors.

Sino 0.5 or 2 mg/kg iv reduced significantly heart rate ( HR), systolic blood pressure ( SBP ), diastolic blood pressure ( DBP ), left ventricular systolic pressure ( LVSP) , dP/dtmax, cardiac index ( CI ) and total peripheral resistance ( TPR ) . VCE- + dP/dtmax was unchanged. T value did't change in 0 .5mg/kg, but it elevated in 2 mg/kg.

Microbiome dysbiosis is strongly associated with alcoholic liver disease(ALD).Recent studies on comprehensive analyses of microbiome compositional and functional changes have begun to uncover the mechanistic relation between microbiome and the pathogenesis of ALD.Importantly,targeting the microbiome has become a potential strategy for the prevention and treatment of ALD.In this review,we summarize the clinical evidence of microbiome dysbiosis in ALD patients,and experimental advances in microbiome and metabolomic functional changes in animals with different species and genetic back-grounds in ALD.We also summarize the studies in humanized intestinal microbiome and fecal micro-biota transplantation in mice.We introduce new developments in the studies on the role of the circulating bacterial microbiome,oral bacterial microbiome and fungal microbiome in the development of ALD.We highlight the potential mechanisms by which microbiome dysbiosis contributes to ALD,including short chain fatty acid changes,bile acid metabolism,intestinal barrier function,release of bacterial and fungal products,and inflammation.In addition,we summarize the recent developments targeting the microbiome in prevention and treatment of ALD,including dietary nutrient interference,herbal medicine,antibiotics,anti-fungal agents,probiotics,engineered bacterial therapy,fecal trans-plantation and oral hygiene.Although recent preclinical studies have advanced our understanding of the microbiome and ALD,clinical studies,especially prospective studies with large samples,are needed to better understand the cause-effect of microbiome dysbiosis in ALD.Identifying new precision-based strategies targeting the microbiome are expected to be developed as more effective therapies in ALD.

Objective:To determine the dosage,method and effectiveness of using magnesium sulfate for treating the intractable epilepsy.Methods:72 cases were allocated to treatment group and 36 to control group. For those in the treatment group,either 10 mL of 25% MgSO4 was administered by intravenous drip or 10 mL of 16.5% MgSO4 by oral intake once per day,in addition to the use of the commonly used antiepilepsy drugs. For those in the control group,the commonly used antiepilepsy drugs such as tegretol,sodium phenytoin,luminal or γ-Aminobutyric Acid were employed.Results:The improvement rate in the treatment group was 87.50%,higher than that (44.44%) in the control group (P＜0.01).Conclusion:Using magnesium sulfate in conjunction with the antiepilepsy drugs is a simple yet effective regime for the treatment of the intractable epilepsy.

BACKGROUND:Bone marrow mesenchymal stem cel s from chickens are important cel models for embryonic developmental biology, immunology and oncology research. However, it is difficult to keep bone marrow mesenchymal stem cel s with good undifferentiated potential in a large-scale expansion system.
OBJECTIVE:To establish a culture system in vitro with laminin coating to expand bone marrow mesenchymal stem cel s from chickens.
METHODS:Isolated bone marrow mesenchymal stem cel s from chickens were seeded in laminin-coated plates and traditional two-dimensional plates, respectively. After expansion in vitro, the morphological characteristics, expression of surface markers, expansion characteristics and adipogenic differentiation of bone marrow mesenchymal stem cel s in both conditions were analyzed and compared.
RESULTS AND CONCLUSION:There were no statistical differences in the morphological characteristics and expression of surface markers of bone marrow mesenchymal stem cel s expanded by laminin-coated plates and traditional two-dimensional plates. But, the expansion characteristics and adipogenic differentiation of bone marrow mesenchymal stem cel s cultured in laminin-coated plates were better than those in traditional two-dimensional plates. Laminin culture system could quickly amplify out of a large number of chicken bone marrow mesenchymal stem cel s with better proliferation ability and undifferentiated performance. Al above results indicated that a more efficient expansion system with laminin coating is established.

Objective To analyze the clinicopathologic factors affecting the formation of lymphovascular invasion (LⅥ) in patients with gastric cancer.Methods The retrospective case-control study was conduted.The clinicopathologic data of 1 260 patients with gastric cancer who were admitted to the First Affiliated Hospital of Nanjing Medical University between January 2014 and December 2015 were collected.All the surgical specimens of patients were detected by hematoxylin-eosin (HE) stain and diagnosed by pathological experts.Stages of patients were evaluated by the seventh TNM staging system for gastric cancer of American Joint Committee on Cancer (AJCC) and Union for International Cancer Control (UICC).Observation indicators:(1) pathologica features:histological differentiation,invasive depth,lymph node metastasis and TNM staging;(2) follow-up situations;(3) influenced factors of the positive LⅥ:sex,age,histological differentiation,invasive depth,number of lymph node metastasis and TNM staging affecting positive LⅥ were analyzed.Follow-up using outpatient examination and telephone interview were performed to detect survival of patients up to June 2016.Univariate analysis was done using the chi-square test,and multivariate analysis was done using the trend chi-square test,and binary Logistic regression model.Results (1) Pathological features:1 260 patients with gastric cancer were diagnosed by postoperative pathological examinations,including 355 with positive LⅥ and 905 with negative LⅥ.Histological differentiation:high-differentiated tumor was detected in 13 patients,moderate-differentiated tumor in 232 patients and low-differentiated tumor in 775 patients.There were 95 patients with mucinous adenocarcinoma and 145 with signet-ring cell carcinoma.Invasive depth:tumor invasion into mucosal layer or submucosal layer (T1 stage) was detected in 242 patients,muscular layer (T2 stage) in 160 patients,gastric wall layer and no invasion into serosal layer (T3 stage) in 37 patients and subserosal layer (T4 stage) in 821 patients.Lymph node metastasis:no regional lymph node metastasis (N0 stage) was detected in 461 patients,1-2 lymph nodes metastases (N1 stage)in 164 patients,3-6 lymph nodes metastases (N2 stage) in 245 patients and more than 7 lymph nodes metastases (N3 stage) in 390 patients.TNM staging:there were respectively 191 patients in Ⅰ A stage,114 in Ⅰ B stage,62 in ⅡA stage,202 in ⅡB stage,132 in ⅢA stage,80 in ⅢB stage,476 in ⅢC stage and 3 in Ⅳ stage.(2)Follow-up situations:1 142 patients (320 with positive LⅥ and 822 with negative LⅥ) were followed up for 4.0-24.0 months,with a meidan time of 11.0 months and a follow-up rate of 90.635％ (1 142/1 260).During the follow-up,154 patients died,including 41 with positive LⅥ and 113 with negative LⅥ.(3) Influenced factors of the positive LⅥ:① results of univariate analysis showed that histological differentiation,invasive depth,number of lymph node metastasis and TNM staging were factors affecting positive LⅥ of patients with gastric cancer (X2=16.930,29.190,64.463,46.539,P＜0.05).② Results of the trend chi-square test showed that histological differentiation,invasive depth,number of lymph node metastasis and TNM staging were factors affecting positive LⅥ of patients with gastric cancer,with a linear correlation (X2 =54.883,69.130,164.618,119.594,r=0.211,0.243,0.365,0.316,P＜0.05).There was a greater correlation between number of lymph node metastasis and formation of lymphovascular invasion.③ Results of the binary Logistic regression model showed that moderate-and low-differentiated tumor and N1-N3 stage of lymph node metastasis were independent risk factors affecting positive LⅥ of patients with gastric cancer (OR=2.572,1.782,95％ confidence interval:0.495-1.494,0.386-0.781,P＜0.05).Conclusion Patients with lower tumor differentiation and / or greater number of lymph node metastasis may have a higher risk of forming LⅥ.

Purpose　To investigate the clinicopathological and immunohistochemical features of Epstein-Barr virus-associated and Epstein-Barr virus not-associated primary intestinal T-cell lymphomas(ITCL) and to study their cell origins. Methods　In situ hybridization for EBER1/2 and immunohistochemical staining for immunophenotypes, LMP-1,TIA-1,bcl-2 and CD21 were performed in 32 cases. The clinical data were analyzed and all patients were followed-up. Results　(1) In 27 of the 32 cases, EBER1/2 were detected in the tumor cells, in which 11 presented LMP-1 positive reactions. (2) All 32 cases of ITCL revealed CD45RO positivity，in which 4（12.5%） expressed CD8，8（25.0%）expressed CD4, 9（28.1%）expressed CD56,and 31（96.9%）expressed TIA-1. There were 17（53.7%）cases with CD4-，CD8-，CD56- immunophenotype. None expressed bcl-2 and CD21. 32 ITCL were classified into pleomorphic medium and large cell(n=28), monomorphic medium-sized(n=2), pleomorphic small cell(n=2). Clinically, most patients with ITCL were young males with abdominal pain, hematochezia, fever and weight loss. The prognosis of patients with ITCL showed poor (survival median was 1.7 month). (3) The differences between EBV-associated and EBV not-associated ITCL lay in hematochezia, fever and the expression of CD3, CD8 and CD56. Conclusion　Most of Chinese ITCL are EBV-associated ones with unusual clinicopathological and immunohistochemical features，which are of different lineages of T-cell subtypes, including cytotoxic T-cell or NK cell.

Objective To investigate the expressions of survivin and Ki67 in cutaneous extranodal NK/T-cell lymphoma,nasal type and their significance.Methods Clinical data and laboratory test results were collected from 15 cases of cutaneous extranodal NK/T-cell lymphoma,nasal type with skin lesions as the initial manifestation.Immunohistochemical SP method was used to measure the expression of survivin and Ki67 in tissue sections from the lesions of these patients.To determine the percentage of survivin-or Ki67-positive cells,200 tumor cells were counted in a high power field (HPF) and 5 HPFs were observed in 1 section.Results There were 10 males and 5 females among the 15 patients,with the median age at diagnosis being 28.7 (range:9-62) years.Immunohistochemical study showed that the lesional tissue was positive for CD56 in 9 cases,CD3ε in 13 cases,T cell intracellular antigen (TIA)-1 in 15 cases,granzyme B in 10 cases,CD3 in 2 cases,βF1 in 1 case,but negative for CD4,CD5,CD8,CD20,and CD79α.All the 15 cases were positive for 1 or 2 T-cell associated antigens (CD2,CD7 or CD45RO).CD30 was observed in large tumor cells in 3 of the 15 cases.Monoclonal TCR-γ gene rearrangement was found in 3 of 14 cases.In situ hybridization for EBER1/2 was positive in all of the 15 cases.Survivin and Ki67 were expressed in 73.3％ (11/15) and 100％ (14/14,note:a tissue section was unavailable because of the detachment of tissue from the slide) of the specimens respectively,with the average percentage of positive cells being 23.97％ ± 18.35％ and 41.20％ ± 19.52％ for survivin and Ki67,respectively.Statistical differences were observed in the percentage of Ki67-expressing cells between the specimens with 0-2 mitotic figures per high power field from 9 cases and those with more than 2 mitotic figures from 6 cases (25.27％ ± 12.96％ vs.58.23％ ± 16.02％,F =19.14,P =0.001).No correlation was found between the expression of survivin and Ki67 in cutaneous extranodal NK/T-cell lymphoma,nasal type.Conclusion The high expressions of survivin and Ki67 may play a role in the occurrence and development of cutaneous extranodal NK/T-cell lymphoma,nasal type.