目的：建立存活时间较长的急性重症肝炎小鼠模型。方法：将50只BALB/c小鼠平均分成5组，其中A、B、C、D 4组为实验组，E组为对照组；实验组分别给予D氨基半乳糖（D-GalN）和脂多糖（LPS），剂量分别为800 mg/kg和10 μg/kg、500 mg/kg和10 μg/kg、500 mg/kg和5 μg/kg、300 mg/kg和5 μg/kg，用生理盐水稀释至1 mL，行腹腔注射；对照组E腹腔注射生理盐水2 mL。以12 h死亡率、24 h死亡率及肝组织学改变为观察指标。结果：A、B、C、D及E组小鼠12 h死亡率分别为80%%、30%、10%、0和0；24 h死亡率分别为90%、60%、30%、0和0；A和B组小鼠肝组织学均呈急性重症肝炎表现，C组中有5只小鼠肝组织学符合重症肝炎的表现，而该组其余小鼠及D组所有肝组织学虽有炎症改变，但达不到重症肝炎的程度，E组肝组织学表现正常。结论：LPS 10 μg/kg联合D- GalN 500 mg/kg可成功建立12 h存活率较高的急性重症肝炎小鼠模型。
目的：建立能表达乙肝病毒（hepatitis B virus,HBV）preS2S抗原蛋白的荷瘤小鼠模型，为研究HBV核酸疫苗的体内CTL应答及免疫治疗作用提供简便易行的研究模型。 方法：以免疫印迹法验证SP2/0-S2S细胞中有HBV preS2S抗原的稳定表达，将SP2/0-S2S细胞种植到BALB/c小鼠胁部皮下（荷瘤），观察能否生长成瘤，以及成瘤的时间、肿瘤大小和荷瘤后小鼠的生存时间；以免疫组织化学法检测小鼠肿瘤组织HBV preS2S抗原的表达。以不表达preS2S抗原蛋白的SP2/0-CMV细胞荷瘤小鼠作为阴性对照。结果：荷瘤后3天～1周，SP2/0-S2S细胞可在小鼠皮下形成实体肿瘤，成瘤率为100%，肿瘤细胞中有preS2S抗原表达，荷瘤后小鼠的平均生存时间为16±1天；与不表达preS2S抗原蛋白的SP2/0-CMV细胞荷瘤小鼠相比，成瘤率、成瘤时间、肿瘤大小及生存时间差异。结论：建立了能表达HBV preS2S抗原蛋白的荷瘤小鼠模型，可用于HBV核酸疫苗的体内CTL应答及免疫治疗作用的实验研究。同时也建立了不表达preS2S抗原蛋白的荷瘤小鼠模型，可用作阴性对照。
ObjectiveTo establish a mouse model of acute necrotizing pancreatitis.MethodsThirty-six male ICR mice were randomly divided into control group (n=6) and experimental group (n=30). Each of the animals in the experimental group received 7 intraperitoneal injections of caerulein (50 μg/kg body weight) in 0.9% NaCl at hourly intervals over 6 hours. The animals in the experimental group were killed at 9,18,24,48 and 72 hours respectively after the first caerulein injection. The control animals received the same volume of 0.9% NaCl without caerulein. The animals in the control group were killed at the 18th hour after the first intraperitoneal injection. The severity of acute necrotizing pancreatitis was evaluated in terms of amylase level, pancreatic weight/body weight and the histological changes. Variance analysis was employed in the processing of these data. ResultsBoth amylase level and pancreatic weight elevated 9 hours after the first caerulein injection, and correlated with the course of pancreatitis. The maximums of both alterations were observed at the same time point (18 hours after the first injection of caerulein). Prominent interstitial inflammation and acinar cell necrosis occurred at the 18th hour, and the histological score for pancreatitis reached a maximum （P＜0.05）. Conclusion Intraperitoneal injection of a large dosage of caerulein can induce acute necrotizing pancreatitis in ICR mice. This method is simple and noninvasive, and the model established thus is stable and reproducible.
ObjectiveTo establish an animal model of anaplastic thyroid cancer with high metastatic activity as in human body. MethodsHuman anaplastic thyroid cancer cell line TAK was injected into one of the lateral lobes of the thyroid gland, as well as in the subcuitis in a series of nude mice. Mice were sacrificed when found moribund, and autopsy and histology were performed subsequently.ResultsThe implantation of human anaplastic thyroid cancer cells in an ectopic enviroment did not permit expression of metastasis potential. In contrast, intrathyroid implantation did. Lymph node (5/10), lung (3/10) and one metastasis (1/10) were noted upon histological examination. ConclusionAn animal model with high metastatic activity is established when human anaplastic thyroid cancer cell line TAK is implanted orthotopically into nude mice.
In this study, the role of newcastle disease virus (NDV) combined thermic solidified tumor vaccine in inhibiting growth of tumor and immune control was investigated, and rate of inhibiting tumor and cellular immunity were measured. The results showed that rate of inhibiting tumor in experimental group Ⅰ and Ⅱ were 24.8% and 41.1% respectively; average weight of tumor was significantly lower in both experimental groups than in control group, and activity of natural killing (NK) cells in experimental groups was higher than that in control group (Ｐ＜0.01). This suggests that NDV combined thermic solidified tumor vaccine can inhibit growth of tumor and improve activity of NK cells, and their effects are better than that of NDV.
On the basis of established JF305 cell line from human pancreatic cancer at this university, cell clone technique, cell electrophoresis, flower cytometer, and cancer orthotopically implanted nude mice technique were used to establish the sublines with different metastatic potential from human pancreatic cancer line-JF305 and the nude mice model implanted orthotopically with human pancreatic cancer monoclonal sublines with different metastatic potential. The results showed that the monoclonal cell sublines with different metastatic potential from human pancreatic caner-JF305 and the nude mice model implanted orthotopically with the sublines, would provided a useful method to study the metastatic mechanism of human pancreatic cancer.
Ｉｎ ｏｒｄｅｒ ｔｏ ｒａｉｓｅ ｔｈｅ ｅｆｆｉｃｉｅｎｃｙ ｏｆ ｃｈｅｍｏｔｈｅｒａｐｙ ｏｎ ｍａｌｉｇｎａｎｔ ｔｕｍｏｒ，ｗｅ ｔｒｅａｔｅｄ ｔｒａｎｓｐｌａｎｔｅｄ ｈｅｐａｔｏｃａｒｃｉｎｏｍａｂｅａｒｉｎｇ ｍｉｃｅ ｗｉｔｈ Ｇｅ１３２（ｏｒｇａｎｉｃ ｇｅｒｍａｎｉｕｍ）．Ｔｈｅ ｒｅｓｕｌｔｓ ｗｅｒｅ∶① ｉｎ ｔｈｅ ｇｒｏｕｐ ｏｆ ｈｉｇｈｄｏｓａｇｅ Ｇｅ１３２，ｔｈｅ ｃａｒｃｉｎｏｓｔａｔｉｃ ｒａｔｅ ｗａｓ ３１０％，＋＋＋ ｗｈｉｔｅ ｂｌｏｏｄ ｃｅｌｌ（ＷＢＣ）ｉｎｆｉｌｔｒａｔｉｏｎ ｆｏｕｎｄ ｉｎ ｔｒａｎｓｐｌａｎｔｅｄ ｔｕｍｏｒ ｗａｓ ７２７％，ａｎｄ ｔｈｅ ｓｅｒｕｍ ｓｕｐｅｒｏｘｉｄｅ ｄｉｓｍｕｔａｓｅ （ＳＯＤ） ａｃｔｉｖｉｔｙ ｒａｔｅ ｗａｓ １２１ Ｎｕ／ｍｌ； ② ｉｎ ｔｈｅ ｃｏｎｔｒｏｌ ｇｒｏｕｐ ｗｉｔｈ ｎｏｒｍａｌ ｓａｌｉｎｅ ｓｏｌｕｔｉｏｎ （ＮＳ），＋＋＋ ＷＢＣ ｉｎｆｉｌｔｒａｔｉｏｎ ｆｏｕｎｄ ｉｎ ｔｕｍｏｒ ｗａｓ ３６．４％， ｔｈｅ ｓｅｒｕｍ ＳＯＤ ａｃｔｉｖｉｔｙ ｒａｔｅ ｗａｓ ８１ Ｎｕ／ｍｌ，ａｎｄ ｔｈｅｒｅ ｗａｓ ｓｉｇｎｉｆｉｃａｎｔ ｄｉｆｆｅｒｅｎｃｅ （Ｐ＜０．０５） ｂｅｔｗｅｅｎ ｔｈｅ ｔｗｏ ｇｒｏｕｐｓ；③ ｉｎ ｇｒｏｕｐ ｏｆ ｃｙｃｌｏｐｈｏｓｐｈａｍｉｄｅ ａｄｍｉｎｉｓｔｒａｔｉｏｎ，ｔｈｅ ｃａｒｃｉｎｏｓｔａｔｉｃ ｒａｔｅ ｗａｓ ３７．０％，＋＋＋ ＷＢＣ ｉｎｆｉｌｔｒａｔｉｏｎ ｆｏｕｎｄ ｉｎ ｔｈｅ ｔｕｍｏｒ ｗａｓ ２７．３％，ａｎｄ ｔｈｅ ｓｅｒｕｍ ＳＯＤ ａｃｔｉｖｉｔｙ ｒａｔｅ ｗａｓ １０２ Ｎｕ／ｍｌ；ａｎｄ ④ ｈｉｇｈｄｏｓａｇｅ Ｇｅ１３２ ｉｎ ｃｏｍｂｉｎａｔｉｏｎ ｗｉｔｈ ｃｙｃｌｏｐｈｏｓｐｈａｍｉｄｅ，ｓｈｏｗｅｄ ａ ｃａｒｃｉｎｏｓｔａｔｉｃ ｒａｔｅ ｏｆ ４５．０％，＋＋＋ ＷＢＣ ｉｎｆｉｌｔｒａｔｉｏｎ ｆｏｕｎｄ ｉｎ ｔｕｍｏｒ ｗａｓ ５４．５％，ｔｈｅ ｓｅｒｕｍ ＳＯＤ ａｃｔｉｖｉｔｙ ｗａｓ １４２ Ｎｕ／ｍｌ．Ｉｎ ｃｏｍｐａｒｉｓｏｎ ｂｅｔｗｅｅｎ ｇｒｏｕｐ ２ ａｎｄ ５，ｔｈｅｒｅ ｗｅｒｅ ｓｉｇｎｉｆｉｃａｎｔ ｄｉｆｆｅｒａｎｃｅ （Ｐ＜０．０５） ｉｎ ｂｏｔｈ ｔｉｓｓｕｅ ＷＢＣ ｉｎｆｉｌｔｒａｔｉｏｎ ａｎｄ ＳＯＤ ａｃｔｉｖｉｔｙ．Ｔｈｅｓｅ ｒｅｓｕｌｔｓ ｓｕｇｇｅｓｔ ｔｈａｔ ｈｉｇｈｄｏｓａｇｅ Ｇｅ１３２ ｔｒｅａｔｍｅｎｔ ｃａｎ ｅｎｈａｎｃｅ ｔｈｅ ｃｅｌｌ ｍｅｄｉａｔｅｄ ｉｍｍｕｎｉｔｙ ａｎｄ ａｃｔｉｖｉｔｙ ｏｆ ｓｅｒｕｍ ＳＯＤ ｏｆ ｔｒａｎｓｐｌａｎｔｅｄ ｈｅｐａｔｏｃａｒｃｉｎｏｍａｂｅａｒｉｎｇ ｍｉｃｅ ｗｉｔｈ ｃｅｒｔａｉｎ ａｎｔｉｃａｒｃｉｎｏｍａ ｅｆｆｅｃｔ．
Objective To investigate the gene expression of beta-defensin-4 (mBD-4) and mBD-6 in acute lung injury (ALI) mouse.Methods Sixty adult mice were randomly divided into a control group and a ALI group.ALI was induced by intraperitoneal injection of lipopolysaccharide (LPS) in the ALI group.The control group was treated with same dose of normal saline.The lung tissues were harvested at different time point after stimulation.The expression of mBD-4 and mBD-6 mRNA was measured by real-time quantitative reverse transcription polymerase chain reaction.DNA sequencing was used to confirm the specificity of mBD-4 and mBD-6 cDNA fragment.Results There were no obvious mBD-4 and mBD-6 mRNA expression in mouse lung in the control group at all time points and ALI 6 h group.In the ALI group a marked increasing expression was found on 12 h，1 d and 3 d after LPS stimulation.The mBD-4 mRNA expression was significant higher in the ALI groups of 1 d and 3 d points than that of ALI 12 h group with no obvious difference between each other.There were no significant differences of mBD-6 mRNA expression between ALI groups of 12 h，1 d and 3 d points Conclusion mBD-4 and mBD-6 mRNA is not constitutive expressed in mouse lung and show a up-regulative expression pattern after ALI.
Objective To observe the expression of adenovirus vector coding for mouse endostatin gene（Ad-mES） in lung cancer cells and its antiangiogenic activity in human umbilical vein endothelial cells（ECV304） in vitro.Methods Lewis lung cancer（LLC） cells were transfected with Ad-mES at different multiplicity of infection（MOI）.The expression of mES in LLC cells and supernatant after 48 hours was detected by immunohistochemical staining and Western blot respectively.The inhibitory effect of supernatant at different MOI on ECV304 non-stamulated and stimulated by basic fibroblast growth factor（bFGF） was measured by methyl thiazolyl tetrazolium（MTT） assay.Results After transfected for 48 hours，endostatin was identified in the cell plasma of infected LLC and negative result was founded in non-infected LLC.Western blot revealed band of endostatin in 20 kDa in culture supernatant of infected LLC and negative results in non-infected LLC.The inhibitory effects on ECV304 cell proliferation were ber at higher MOI，and the difference was significant between stimulated and non-stamulated cells by bFGF（Plt;0.05）.Conclusion Ad-mES can transfect and express endostatin effectively in LLC with biological activity
Objective To explore whether epithelial to mesenchymal transition ( EMT) occurs in bleomycin( BLM) induced pulmonary fibrosis, and the involvement of bronchial epithelial cells( BECs) in the EMT. Methods BLM-induced peribronchial fibrosis in an α-smooth muscle actin-Cre transgenic mouse( α-SMACre /R26R) was examined by pulmonary βgal staining and α-SMA immunofluorescence staining. Results BLMtreated mice showed significantly enhanced βgal staining in subepithelial areas in bronchi, terminal bronchioles and walls of pulmonary vessels. Some alveolar epithelial cells( AECs) in certain peribronchial areas or even a small subset of BECs were also positively stained, as confirmed by α-SMA immunostaining. Conclusions EMT occurs in BLM-induced peribronchial fibrosis mice. BECs, like AECs, have the capacity to undergo EMT and to contribute to mesenchymal expansion in pulmonary fibrosis.