1.Inhibitory effect of dendritic cells combined with cytokine-induced killer cells on the growth and liver metastasis of human primary malignant melanoma of the small intestine
Shuai TUO ; Ningsu ZHANG ; Ning ZHANG
Chinese Journal of Digestive Surgery 2010;09(6):426-429
Objective To investigate the inhibitory effect of dendritic cells (DCs) and homologous cytokine-induced killer cells (CIKs) on the growth and liver metastasis of human primary malignant melanoma of the small intestine. Methods A histologically intact liver metastasis fragment derived from a surgical specimen of patients with primary malignant melanoma of the small intestine was implanted in the mucous layer of small intestine in nude mice to construct the liver metastasis model of the malignant melanoma. Peripheral blood mononuclear cells from healthy donors and patients with malignant melanoma of the small intestine were isolated. DCs and CIKs were separately produced according to corresponding culture method, and the DC-CIKs were harvested after coculorthotopically implanted in 56 nude mice, and 72 hours later, all the mice were equally divided into normal saline group (0.3 ml), mitomycin treatment group (400 μg/0.3 ml), healthy donor CIK group (6 × 107/0.3 ml),healthy donor DC-CIK group (3 × 107/0.3 ml), autologous CIK treatment group (6 × 107/0.3 ml), autologous DC-CIK group (6 × 107/0. 3 ml) and DC-CIK (6 × 107/0.3 ml) + mitomycin (400 μg/0. 3 ml) treatment group. Mice in the experimental groups were administered the agents once daily by vena caudalis injection for 28 days. Mice were sacrificed 72 hours after treatment, and the orthotopic xenografts were removed for weighing,and the liver metastases were simultaneously evaluated by macroscopy and microscopy. All data were analyzed using the analysis of variance, LSD test and chi-square test. Results A liver metastatic model of human primary malignant melanoma of the small intestine ( HSIM-0603 ) was established. Of the control group, healthy donor CIK group, healthy donor DC-CIK group, autologous CIK treatment group, autologous DC-CIK group and DC-CIK +mitomycin group, the tumor weights were ( 1.18 ± 0. 17), (0.71 ± 0.06), (0.68 ± 0. 15 ), (0. 43 ± 0.08 ),(0.67 ± 0.07 ), (0.37 ± 0.08 ) and (0.20 ± 0.05 ) g, respectively; the tumor inhibition rates were 0, 39.82%,42.37%, 65.56%, 43.22%, 68.64% and 83.05%, respectively; liver metastases were detected in 8, 8, 5,4, 4, 3, 2 rats, respectively, in the above mentioned groups. There were significant differences in the tumor weight, tumor inhibition rate and liver metastasis rate among the seven groups (F = 152. 300, x2= 200. 538,94. 325, P < 0.05 ). Conclusions The liver metastatic model could be applied to the studies on the pathogenesis,invasion, metastasis and treatment of human primary malignant melanoma of the small intestine. DC-CIK has the potential in inhibiting the growth and liver metastasis of the malignant melanoma of the small intestine.
2.Reproduction of a murine model of liver metastasis of human primary malignant lymphoma of small intestine by continuous screening in vivo
Chaowei TUO ; Qiuzhen LIU ; Ning ZHANG
Medical Journal of Chinese People's Liberation Army 2001;0(09):-
Objective A liver metastasis model of human primary malignant lymphoma of small intestine was reproduced in nude mice in an attempt to provide an ideal animal model for elucidating the mechanism of liver metastasis of primary small intestinal lymphoma.Methods A piece of surgically obtained liver metastatic tissue of small intestinal lymphoma was implanted into the mucous membrane of small intestine in nude mice to reproduce the model.After metastasis of this tumor to the liver occurred,a portion of the metastais was transplanted into the mucous membrane of small intestine of another nude mouse.This process was repeated 4 times in order to obtain a cell line with the characteristics of high malignant lymphoma metastasis to liver.The survival rate of the experimental animals,regional invasion rate and metastasis rate were observed,and the morphological characteristics(light microscopy,electron microscopy and immunohistochemistry),karyotype analysis and DNA content of the neoplastic cells were also determined.Results A liver metastasis model of human primary malignant lymphoma of small intestine in nude mice(termed HSIL-0402)was successfully reconstructed,by repeated implantation of liver metastatic tumor in vivo.Histopathology showed HSIL-0402 tumor was a high grade non-Hodgkin's lymphoma.Immunohistology showed the cells were CD19,CD20,CD45 and CD79a positive,but negative for CD3 and CD7.The modal number of chromosome was between 56-69.DNA index(DI)was 1.61?0.37,which showed heteroploid.So far,HSIL-0402 had been maintained for 27 passages in nude mice,exhibiting 100% of transplantability,liver metastatic rate,and resuscitation rate after liquid nitrogen cryopreservation.Lymph node metastasis and celiac seeding occurred in 67.4% and 61.7% of a total of 156 observed animals.The HSIL-0402 model displayed various manifestations reminiscent of highly metastatic invasive behavior in nude mice,including invasive growth,hematogenous metastasis,lymph node metastasis and celiac seeding.Conclusion The present study successfully re-established a spontaneous liver metastasis model of human primary malignant lymphoma of small intestine in nude mice,HSIL-0402,which provides an ideal animal model for the researches on biological mechanism of liver metastasis and anti-metastasis therapy of human primary small intestinal lymphoma.
3.Inhibitory effect of CIK cells on growth and liver metastasis of human primary gastric malignant lymphoma xenograft
Bo YANG ; Chaowei TUO ; Ning ZHANG
Medical Journal of Chinese People's Liberation Army 2001;0(11):-
Objective Liver metastasis model of human primary gastric lymphoma in nude mice was reproduced for an experiment to evaluate the inhibitory effect of cytokine-induced killer (CIK) cells on the growth and liver metastasis of primary gastric lymphoma. Methods Surgical orthotopic implantation of a histologically intact liver metastasis fragment derived from a surgical specimen of a patient with metastatic gastric lymphoma was initally implanted, in order to reprodueing a liver metastasis model of human primary gastric lymphoma. Peripheral blood mononuclear cells (PBMC), isolated by blood cell separator from healthy donors and patients with primary gastric lymphoma, were incubated in vitro. rhIFN-?, rhIL-2 and anti-CD3 McAb were added to PBMC in order to prepare CIK cells as well as lymphokine-activated killer (LAK) cells. CIK and LAK cells from different donors were used in treating gastric malignant lymphoma, so as to investigate the inhibitory effect of 2 kinds of effector cells on the growth of the tumor and liver metastasis. Results The liver metastasis model of human primary gastric malignant lymphoma in nude mice was successfully reproduced. After administration of different agents continuously for 20 days (0.3ml/d), the inhibitory rates of the following 4 groups, healthy donors LAK group (2?1010/ml), healthy donors CIK group (2?1010/ml), patients CIK group (1?1010/ml) and patient CIK group (2?1010/ml), were 39.28%, 53.57%, 40.38% and 56.42%, respectively. The liver metastasis rates in control group, healthy donors LAK group, healthy donors CIK group, patients CIK group (1?1010/ml) and patient CIK group (2?1010/ml), were 100.0%, 62.5%, 50.0%, 62.5% and 37.5%, respectively. Tumor weights of all treatment groups were lighter than that of saline group (P
4.THE CHARACTERISTIC OF THE ULTRASTRUCTURE OF VASCULAR AND CONNECTIVE TISSUES IN PRIMARY SYPHILIS
Xiaodong ZHANG ; Caizhong WU ; Chaowe TUO
Medical Journal of Chinese People's Liberation Army 1982;0(01):-
To study the ultrastructural pathological characteristics of vascular and connective tissues in primary syphilis. The histomorphological changes in the dermis were examined with transmission electron microscopy. Capillaries with a thickened multilayered basement membrane,occluded lumen, and proliferation of endothelial cells were found. The spirochetes entered the capillary vessel through the basal membrane and endothelial cells, and were separated or attached to red blood cells.Large numbers of inflammatory cells were found around the blood vessels.There were T.pallidum in the cytoplasm.The pathological changes in the capillaryies and connective tissue were morphological basis for the study of pathogenetic mechanism of primary syphilis.
5.EXPRESSIONS OF CD3 ,CD20,CD68 IN EARLY SYPHILIS LESIONS
Xiaodong ZHANG ; Chaowei TUO ; Fangji SONG
Medical Journal of Chinese People's Liberation Army 2001;0(11):-
To probe course and mechanism of cellular immune respone in early syphilitic lesion, the expression levels of CD3, CD20, CD68 were determined by immunohistochemistry and a computer image analysis system. The results showed that the expressions of CD3, CD20, and CD 68 were positive in 43 cases of primary and secondary syphilis. The expression level of CD68 was higher thanthat of CD3, CD20, especially around blood vessels. The expression level of CD 20 was very low. This study indicates that T cells, B cells, and in particular macrophages play an important cellular immunologic regulating role in clearance of Treponema pallidum from the infected tissue, but the efficacy is not high enough.
6.INFLUENCE OF LACTOBACILLUS CASEI ZHANG ON IMMUNOLOGICAL FUNCTION OF INTESTINAL MUCOSA ASSOCIATED LYMPHOID TISSUES OF MICE
Ruiting DU ; Ya TUO ; Heping ZHANG
Acta Nutrimenta Sinica 1956;0(04):-
Objective To study the effects of L.casei Zhang viable cell preparation on the immune function of small intestinal mucosa in mice. Method L.casei Zhang isolated from Koumiss was administered by gastric intubation to mice in the form of viable organism preparation for 11d and the quantity of Peyer's patch on small intestine ,the CD3+、IgA+ cells number of intestinal mucosal and the average optical density value of their positive signal were observed. Results L casei Zhang could not only promote the number of the Peyer's patch significantly,but also enhanced the quantity and the expression intensity of CD3+,IgA+ cells. Conclusion L.casei Zhang can obviously promote the intestinal mucosa immunity of mice.
7.Reblication of high metastasis model of malignant lymphoma of human small intestine in nude mice
Shuai TUO ; Qiuzhen LIU ; Ning ZHANG
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective To replicate an ideal animal model for exploring pathogenesis and experimental treatment of malignant lymphoma of small intestine. Methods Fresh lymphoma tissues derived from primary lesion and liver metastasis of malignant lymphoma of human small intestine obtained during operation were respectively transplanted into mucosa of small intestine and subcutaneous space in the interscapular region in nude mice. Tumorigenicity, invasion and metastasis of transplanted tumors were observed. Morphology (light microscopy, electron microscopy and immunohistochemistry), karyotype analysis, DNA quantitative assay (FCM) were also studied. Results From five patients of malignant lymphoma of small intestine, tumor tissue from 3 patients was successfully transplanted. According to the new classification of the World Health Organization, a strain of high metastatic model of orthotopically transplanted malignant lymphoma in nude mice (HSIL-0101) and another of subcutaneously transplanted highly metastatic model were reproduced from the same human neoplasm (non-Hodgkin B cell type); orthotopically Pathological study showed that the tumor was high-grade large B-cell lymphoma. Histochemitry showed that it was CD19, CD20, CD22 and CD45 positive, and CD3 and CD7 negative. The number of chromosome is ranged from 55 to 59; DI (DNA Index) was 1.47~1.61 (ie, heteroploid). HSIL-0101 and HSIL-0102 had been passaged for 32 and 38 generations in nude mice separately. 357 nude mice were transplanted. Rate of growth of neoplasm transplantation and resuscitation rate of liquid nitrogen cryopreservation were both 100%. In HSIL-0101, metastasis rate of liver and lymph node was 100%. In HSIL-0102, metastasis rate of liver was 63.5% and metastasis rate of lymph node was 62.7%. Transplanted tumors invasively grew in small intestine and subcutaneous region of nude mice. Blood metastasis was found (liver and spleen metastases). There were also lymph metastasis and seeding metastasis in the peritoneal cavity. Conclusions The study first successfully established spontaneous high metastasis models of malignant lymphoma of human small intestine in nude mice by orthotopic and subcutaneous transplantation. HSIL-0101 and HSIL-0102 could be used to carry out the research on pathogenesis, invasion, metastasis, and experimental therapy of malignant lymphoma of small intestine.
8.Reproduction of a nude mouse model orthotopical transplantation of huma n primary malignant lymphoma of the liver for the study of its biological characteristics
Chaowei TUO ; Qiuzhen LIU ; Ning ZHANG
Medical Journal of Chinese People's Liberation Army 2001;0(10):-
Objective To provide an experimental tool for explo ri ng pathogenesis and experimental treatment strategies for primary lymphoma of th e liver. Methods Histologically intact lymphoma tissues from pa tients with primary lymphoma of the liver were transplanted into hepatic parench yma of nude mice. Tumorgenecity, invasion, metastasis, morphological characteris tics(light microscopy, electron microscopy and immunohistochemistry), serologica l test(AFP, HBsAg and LDH), karyotype analysis and DNA content of orthotopically transplanted tumors were studied. Results Orthotopically trans planted model of human primary malignant lymphoma of the liver in nude mice(desi gnated as HLBL-0102) was successfully reproduced. Histopathology of transplante d tumors showed primary lymphoma of the liver(non-Hodgkin's, B cell). The cells were positive for CD19,CD20,CD45RO and CD79a, but negative for CD3 and CD7. Ser ological tests showed that the serum was AFP negative and HBsAg positive, and t he concentration of LDH was 1267.5U/L. The number of chromosome was between 55 and 59. DNA index(DI) was 1.57~1.61(i.e. heteroploid). So far, the strain HLB L-0102 has grown for 3 years and been passaged for 37 generations in nude mice. Altogether 283 nude mice were used for transplantation. The growth rate and res uscitation rate of liquid nitrogen cryopreservation of transplanted tumors were both 100%. The transplanted tumors autonomoasly and invasively grew in the live r of nude mice, damaging adjacent liver tissues, bile ducts, and veins in the po rtal area. There was no involvement of other tissues and organs or distal lymph nodes. Orthotopically transplanted tumors were consistent in histopathological a nd ultrastructural features, DNA content and chromosomal karyotype with the orig inal human tumor. Conclusions The study is the first attempt to successfully reproduce orthotopically transplantation model of human primary ma lignant lymphoma of the liver in nude mice. HLBL-0102 completely replicates th e natural clinical process of primary lymphoma of the liver, and provides an ide al animal model for further research on the biology and experimental therapeutic strategies of primary lymphoma of the liver.
9.Establishment of high metastasis models of human primary colorectal lymphoma in nude mice by orthotopic transplantation
Qiuzhen LIU ; Chaowei TUO ; Ning ZHANG
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
Objective To reproduce high metastasis models of human primary colorectal lymphoma in nude mice by orthotopic transplantation, and to investigate their biologic features. Methods Histologically fresh lymphoma tissues from primary and liver metastatic lesions of human primary colorectal lymphoma obtained during operations were transplanted into colorectal mucosa of nude mice. Tumorgenecity, invasion, metastasis, morphology (light microscopy, electron microscopy and immunohistochemistry), karyotype analysis and DNA content of the orthotopically transplanted tumors were studied. Results According to the new WHO classification of malignant lymphoma, a strain of liver metastasis model of human primary colonic lymphoma (non-Hodgkin's, B cell) in nude mice by orthotopic transplantation (HCBL-0301), and a strain of high metastasis model of human primary rectal lymphoma (non-Hodgkin's, B cell) in nude mice by orthotopic transplantation (HRBL-0302) were successfully screened from four cases of human primary colorectal lymphoma. Histopathology of transplanted tumors showed high grade non-Hodgkin's large B cell lymphoma. The cells were positive for CD19, CD20, CD22 and CD45, but negative for CD3 and CD7. The number of chromosome was between 55 and 69. DNA index (DI) was 1.59~1.71 (i.e. heteroploid). So far, HCBL-0301 and HRBL-0302 had been passaged for 31 and 27 generations in nude mice, respectively, and transplantation was successful in 326 nude mice. Since the third generation, both the growth rate of transplantation and rate of resuscitation after being thawed from liquid nitrogen cryopreservation were 100%. In HCBL-0301, metastasis to the right lobe of liver was most common and metastatic rate was 100%; additionally, rates metastasis to of lymph node and peritoneal seeding were 67.4%. In HRBL-0302, metastasis to the left and right lobes of liver was most common with metastasis rate of 63.7%, and rates of metastasis to lymph node and peritoneal seeding were 56.4%. Transplanted human primary colorectal lymphoma could autonomously and invasively grow in the colorectum of nude mice, with occurrence of hematogenic, lymph node and implantation metastases. Conclusions The study successfully replicated high metastasis models of human primary colorectal lymphoma in nude mice by orthotopic transplantation. HCBL-0301 and HRBL-0302 models can be used in the research on pathogenesis, mechanism of invasion and metastasis and experimental therapy of human primary colorectal lymphoma.
10.Measurement and analysis of 210Pb radioactivity level in outdoor air during spring in Beijing
Shuaimo YAO ; Fei TUO ; Qing ZHANG ; Qiang ZHOU ; Jing ZHANG
Chinese Journal of Radiological Medicine and Protection 2017;37(4):286-289
Objective To measure and analyze the radioactivity level of 210Pb in outdoor air in Beijing in spring.Methods Portable high flow air samplers were used to collect outdoor air at the ground level to analyze the 210Pb radioactivity in the aerosol filter samples using a laboratory-based high purity germanium (HPGe) γ spectrometer.Results The activity concentration of 210Pb outdoors ranged from 267.2 to 1 697.6 μBq/m3,with an average of (878.7 ± 386.7) μBq/m3.Statistical analysis showed that the activity concentrations 210Pb of outdoors varied with variable air quality.Conclusions The activity concentrations of 210Pb outdoors are detectable in Beijing,varying considerably but within the normal range.