83-1除草剂和速杀威杀虫剂的小鼠耳肿致敏试验

本文采用较新的致敏性检测方法小鼠耳肿实验对８３—１除草剂和纯中药杀虫剂速杀威进行了检测。实验共设６个组，包括：速杀威原液，１０％速杀威，０．１６％８３—１，０．３２％８３—１及阴性和阳性对照组。结果显示４个受试物组在致敏后２４小时和４８小时均未观察到致敏作用，说明该受试物在此条件下不具致敏性，同时也验证了小鼠耳肿试验用于有色受试物检测致敏性的优越性。

中药抗肿瘤血管生成的研究现状

近年来许多医学工作者对中药以及中成药在肿瘤治疗中的应用研究的越来越广泛和深入。中药以它资源丰富,副作用少等优点应用于临床,由此中药对各种疾病的治疗也引起了国内外医学界的重视,特别是近年来西药对肿瘤的治疗中放疗、化疗所产生的副作用,给患者带来了巨大的痛苦和不便。医生们便从中药、中成药方面,探索了不少治疗肿瘤的有效处方。尽管在中药抗肿瘤血管生成的相关研究的成果不多,但是从天然植物或中药中提取一些新的抗肿瘤血管生成的药物,现在已经反战成研究肿瘤的突破口。现有的研究主要是从以下两方面入手:

半边旗提取物5F抗肿瘤作用的实验研究

目的:观察半边旗提取物5F体外、体内抗肿瘤活性及其毒性。方法:用MTT法观察5F注射液对SPCA-1,K562瘤株的体外抗肿瘤作用;采用荷瘤小鼠瘤重、抑瘤率检测提取物5F对小鼠移植性肿瘤S180抑制作用;观察5F对荷艾氏腹水癌小鼠的生命延长率;胸腺指数、肝脏指数、脾脏指数、白细胞数等指标观察5F对免疫器官的影响。结果:5F注射液对SPCA-1、K562瘤株有显著的体外抗肿瘤作用;对小鼠肉瘤S180也有抗肿瘤活性;对荷艾氏腹水癌的小鼠生命有延长作用;同时对免疫系统有一定抑制作用。结论:半边旗有效成分5F有一定的体内、体外抗肿瘤活性;对荷瘤小鼠功能有抑制作用。

雷公藤对荷肝癌(H_(22))小鼠的抑瘤作用的实验研究

目的:探讨中药雷公藤治疗实验性肝癌的疗效。方法:建立荷实体型肝癌(H22)小鼠模型,观察经雷公藤治疗对其肿瘤生长的影响。结果:雷公藤能显著抑制肿瘤的生长,其抑瘤率达35.46%。结论:雷公藤有明显的抗肿瘤作用。

健坤胶囊对大鼠、小鼠动物模型抗炎、镇痛和抗菌作用随机平行对照研究

[目的]观察健坤胶囊对大鼠、小鼠动物模型抗炎、镇痛和抗菌作用。[方法]采用小鼠二甲苯耳廓肿胀实验、大鼠角叉菜胶足肿胀实验和大鼠棉球肉芽肿实验,观察健坤胶囊的抗炎作用;采用小鼠醋酸扭体实验观察其镇痛作用;采用体外抑菌实验观察其抗菌作用。[结果]健坤胶囊明显降低小鼠二甲苯耳廓肿胀,抑制大鼠角叉菜胶足肿胀,抑制大鼠棉球肉芽肿,抑制小鼠醋酸扭体反应,体外明显抑制金黄色葡萄球菌、大肠杆菌、枯草杆菌和绿脓杆菌的生长。[结论]健坤胶囊对大鼠、小鼠动物模型具有明显的抗炎、镇痛和抗菌作用。

竹醋液的抗炎活性研究

研究竹醋液以及在不同的条件下减压蒸馏得到的各馏分的抗炎活性.采用二甲苯致小鼠耳肿胀法检测竹醋液和各馏分的抗炎活性,并根据馏分沸点的不同确定其中主要有效组分.竹醋原液和提取出来的各馏分均显示了不同程度的抗炎活性(P<0.01).其中竹醋原液的抗炎效果最好(60.42%),低沸点的醇、酮、酯、吡啶和直链酸类次之(49.24%),酚类再次之(32.73%),苯甲酸类及其衍生物和醛类抗炎效果较弱.竹醋液具有抗炎活性,为其抗炎应用领域奠定实验基础,对竹醋功用开发具有积极意义,其抗炎机理需进一步研究.

c-Met in pancreatic cancer stem cells: Therapeutic implications

Pancreatic cancer is the deadliest solid cancer and currently the fourth most frequent cause of cancer-related deaths. Emerging evidence suggests that cancer stem cells (CSCs) play a crucial role in the development and progression of this disease. The identification of CSC markers could lead to the development of new therapeutic targets. In this study, the authors explore the functional role of c-Met in pancreatic CSCs, by analyzing self-renewal with sphere assays and tumorigenicity capacity in NOD SCID mice. They concluded that c-Met is a novel marker for identifying pancreatic CSCs and c-Met high in a higher tumorigenic cancer cell population. Inhibition of c-Met with XL184 blocks self-renewal capacity in pancreatic CSCs. In pancreatic tumors established in NOD SCID mice, c-Met inhibition slowed tumor growth and reduced the population of CSCs, along with preventing the development of metastases.

Identify lymphatic metastasis-associated genes in mouse hepatocarcinoma cell lines using gene chip

AIM: In order to obtain lymphogenous metastasisassociated genes, we compared the transcriptional profiles of mouse hepatocarcinoma cell lines Hca-F with highly lymphatic metastasis potential and Hca-P with low lymphatic metastasis potential.METHODS: Total RNA was isolated from Hca-F and Hca-P cells and synthesized into double-stranded cDNA. In vitro transcription double-stranded cDNA was labeled with biotin (i.e. biotin-labeled cRNA, used as the probe). The cRNA probes hybridized with Affymetrix GeneChip() MOE430A (containing 22 690 transcripts, including 14 500 known mouse genes and 4 371 ESTs) respectively and the signals were scanned by the GeneArray Scanner. The results were then analyzed by bioinformatics.RESULTS: Out of the 14 500 known genes investigated,110 (0.8%) were up regulated at least 23 fold. Among the total 4 371 ESTs, 17 ESTs (0.4%) (data were not presented) were up regulated at least 23 fold. According to the Gene Ontology and TreeView analysis, the 110genes were further classified into two groups: differential biological process profile and molecular function profile.CONCLUSION: Using high-throughput gene chip method,a large number of genes and their cellular functions about angiogenesis, cell adhesion, signal transduction, cell motility, transport, microtubule-based process, cytoskeleton organization and biogenesis, cell cycle, transcription,chaperone activity, motor activity, protein kinase activity,receptor binding and protein binding might be involved in the process of lymphatic metastasis and deserve to be used as potential candidates for further investigation.Cyclin D1, Fosl1, Hsp47, EGFR and AR, and Cav-1 are selected as the possible candidate genes of the metastatic phenotype, which need to be validated in later experiments.ESTs (data were not presented) might indicate novel genes associated with lymphatic metastasis. Validating the function of these genes is helpful to identify the key or candidate gene/pathway responsible for lymphatic metastasis, which might be used as the diagnostic markers and the therapeutic targets for lymphatic metastasis.

A novel gain of function mutant in C-kit gene and its tumorigenesis in nude mice

AIM： To transfect mutant C-kit cDNA at codon 579 into human embryonic kidney cell line to observe its role in the pathogenesis of gastrointestinal stromal tumor （GIST）. METHODS： Eukaryotic expression vectors of pcDNA3- Kit-NW and pcDNA3-Kit-W were constructed. Then pcDNA3-Kit-NW and pcDNA3-Kit-W plasrnids were transfected into human embryonic kidney cell line by Upofectamine. The resistant clone was screened by G418 filtration and identified by sequencing, Western blotting, and immunocytochemical staining. Human embryonic kidney cells were divided into three groups including pcDNA3-Kit-NW, pcDNA3-Kit-W, and vector control groups. Absorbency value with a wavelength of 574 nm was detected by MTT analysis. Mice were injected with three groups of cells. Volume, mass, and histological examinations of the tumors in different groups were measured and compared. RESULTS： The C-kit gene and mutant C-kit gene were successfully cloned into the eukaryotic expression vector pcDNA3, pcDNA3-Kit-NW and pcDNA3-Kit-W were successfully transfected into human embryonic kidney cell line and showed stable expression in this cell line. Cell proliferating activity had significant differences between pcDNA3-Kit-NW and pcDNA3, pcDNA3-Kit- NW and pcDNA3-Kit-W （P〈0.05）, respectively. Tumors were only observed in nude mice implanted with cells transfected with pcDNA3-Kit-NW. CONCLUSION： Mutation of C-kit gene increases the proliferation activity of human cells and plays an important role in the malignant transformation of GIST.

Flk-1 specific kinase inhibitor SU5416 blocked angiogenesis of Lewis carcinoma in mouse and prolonged the survival

Objective: To reveal the mechanism and effect of SU5416 in the treatment of mouse Lewis cancer in vivo. Methods: Lewis cell was transplanted into groin of C57/B6 mouse by subcutaneous injection, then SU5416 was administrated intraperitoneally to investigate the impact of SU5416 on tumor angiogenesis and growth in vivo. 32 mice were treated with SU5416 at two different doses every day until the end-point. As a control, 8 mice received no treatment and 8 mice were treated with vehicle (DMSO) only after implantation. Results: Median survival in the treated group was statistically longer compared to that in the control groups (P < 0.05) and no significant systemic adverse was observed. Histological analysis of the treated tumors showed an increase in necroses and reduced in angiogenesis compared to the control tumors. Furthermore, the percent of apoptotic cells increased in the treated tumors by FCM, the expressions of VEGF and KDR had no change after SU5416 administration by western blot. Conclusion: SU5416 may be useful therapeutics drug that specifically inhibits the enzymatic activity of KDR kinase and could down regulate the tumor angiogenesis.

Modulation of MMP-2 and TIMP-2 by low dose radiation in mice bearing S180 sarcoma

Objective To investigate the inhibition of low dose radiation （LDR） on S180 sarcomas and its modulation of MMP-2 and TIMP-2 in mice. Methods $180 subcutaneously implanted tumor model mice were randomly divided into two groups： control （N） and low dose radiation （LDR） groups. N mice were sacrificed after 12 h, whereas LDR mice were sacrificed after 12 （LDR-12 h）, 24 （LDR-24 h）, 48 （LDR-48 h）, and 72 （LDR-72 h） h. Thereafter, we measured the tumor volumes. Histopathology was performed, and P-V immunohistochemistry was applied to assess MMP-2 and TIMP-2 expression. Results Compared with the control group, the tumor growth was significantly inhibited in the LDR groups （P 〈 0.05）. MMP-2 expression was considerably reduced in LDR-24h （P 〈 0.05） and LDR-48h （P 〈 0.05）, whereas the change of TIMP-2 was not obvious in the LDR groups （P 〉 0.05） in contrast to that of the control group. Conclusion LDR can effectively suppress the growth of S180 implanted tumors by reducing MMP-2, which is associated with invasion and metastasis.

Serial observations on an orthotopic gastric cancer model constructed using improved implantation technique

AIM:To establish a gastric cancer nude-mouse model with improved orthotopic implantation and investigate its biological characteristics at different time points.METHODS:Human gastric cancer SGC-7901 cell suspensions were injected subcutaneously into a nude mouse to develop solid tumors,and the tumor tissue pieces were implanted under the serous coat.The nude mice were then euthanized in group every two weeks to observe the primary tumor growth and metastases.RESULTS:Within 2-4 wk,there were no obvious chang-es about the primary tumor in stomach.At the sixth week,the primary tumor began to grow fast,resulting in incrassation of the gastric wall and stenosis of the gastric cavity,and metastases into the liver and lymph nodes were detected.The tumor,which compressed the adjacent organs,gradually became bigger and bigger followed by stenosis or vanishment of the gastric cavity from 8 to 12 wk.There were massive metastases,and the rate of metastasis was 58%in lymph nodes,78%in liver,39%in kidney,and 81%in peritoneum or septum.CONCLUSION:A gastric cancer model is established,which can simulate the clinical tumor behavior and provide experimental carrier for clinical trials of gastric cancer treatment.

Effect of early administration of exogenous basic fibroblast growth factor on acute edematous pancreatitis in rats

AIM： To observe the therapeutic effect of early administration of exogenous Basic fibroblast growth factor （bFGF） on acute edematous pancreatitis （AEP） in rats. METHODS： Thirty male Sprague-Dawley rats were randomly divided into three （n = 10）： normal control group （group Ⅰ）, AEP group （group Ⅱ） and AEP with bFGF treatment group （group Ⅲ）. AEP was induced by subcutaneous injection of cerulein （5.5 μg/kg and 7.5 μg/kg） at 1 h interval into rats of groups Ⅱ and Ⅲ. Three hours after induction of AEP, 100 μg/kg bFGF was administrated intraperitoneally for 1h to group Ⅲ rats. For test of DNA synthesis in acinar cells, 5-bromo-2＇-deoxyuridine （BrdU） labeling solution was intraperitoneally injected into the rats of groups Ⅱ and Ⅲ 24 h after bFGF treatment. The changes in serum amylase, lipase, pancreatic tissue wet/dry ratio were detected. RESULTS： In bFGF treatment group, there was a significant decrease in the volume of serum amylase, lipase and the pancreatic wet/dry weight ratio（1383.0 ± 94.6 U/L, 194.0 ± 43.6 U/L, 4.32 ± 0.32） compared to AEP group （3464 ± 223,7 U/L, 456 ±68,7 U/L, 6.89 ± 0,47） （P 〈 0.01）, and no significant difference was found between bFGF treatment and control group （1289 ± 94.0 U/L, 171 ± 23.4 U/L, 4.12 ± 0.26, P 〉 0.05）. The inflammatory changes such as interstitial edema, polymorphonuclear neutrophUs （PMNs） and vacuolization were significantly ameliorated compared to AEP group （P 〈 0.01）. A small number of BrdU-labeled nuclei were observed in acinar cells of AEP rats （1.8 ± 0.3 nuclei/microscopic field, n = 10） while diffuse BrdU-labeled nuclei were found in bFGF-treated rats （18.9 ± 1.4 nuclei/microscopic field, n = 10） （P 〈 0.01）. Immunohistochemical study showed increased DNA synthesis in pancreatic acinar cells. CONCLUSION： Early administration of exogenous bFGF has significant therapeutic effect on cerulein-induced acute edematous pancreatitis in rats. Its mechanism is related to the amelioration of inflammation and facilitation of pancreatic regeneration.

The effect of low dose radiation on cytokine correlated to injury of lungs induced by Bleomycin A5

Objective： The aim of this study was to explore the effect of low dose radiation on cytokine excreted by mice inbreathing of atomization of PYM. Methods： Kunming male mice were randomly divided into three groups： blank group, PYM group （P group）, low dose radiation ＋ PYM group （P ＋ L group）. Mice of P ＋ L group were given whole body low dose radiation 75 mGY, dose rate were 12.5 mGY/min. After 6 h, mice in both P ＋ L group and P group were given inbreathe of atomization of PYM, concentration was 2 mg/mL. Mice were sacrificed after the dl, d7, d14, d21 and d28, IL-6 were detected in alveolar irrigating solution. The tissue samples of mice lung were fixed in 10% formalin, TNF-α and TGF-β were analyzed by immuno- histochemistry. Results： Compared with P group, IL-6 in low dose radiation ＋ PYM group were lower, near to blank group, the difference had notable statistical significance on the dl and the d7, while on the d14, d21, d28, the difference had not statisti- cal significance. It suggested that low dose radiation could reduce the resection of IL-6 at the begin of lung injure induced by low dose radiation. The expression of TGF-β and TNF-α in P ＋ L group were lower than that in P group, but the difference in the two groups had statistical significance by gray analysis P 〈 0.05. Conclusion： In the early stage of lung injure caused by PYM in mice, low dose radiation of 75 mGY can reduce the secretion of IL-6, decrease the production of TGF-β and TNF-α.

Assistant radiotherapeutic effect of Jiaqi Mixture on tumor-bearing mice

Objective:The aim was to study the assistant radiotherapeutic effect of Jiaqi Mixture (JQM) on Ehrlich's ascites carcinoma (EAC) mice.Methods:The EAC-cancer model was made up with Kunming mice.The tumor-bearing mice were treated with whole body exposure (8 Gy) and intragastric administration of JQM,and the changes of tumor weight,the total number of white blood cells (WBC) and immune system were observed.Results:The average tumor weight,WBC,spleen coefficient,the stimulation index (SI) of Con A and LPS and the natural killing (NK) cell activity of mice decreased in some degree after radiotherapy,but the average tumor weight decreased more obviously in radiotherapy + medicine groups (compared with tumor control group,P < 0.05);and the other above indexes were much higher in radiotherapy + medicine groups than those in radiotherapy groups (P < 0.05-0.01).Conclusion:It was suggested that JQM can enhance the effect of radiation therapy and protect the normal immune system caused by radiation therapy.

A New Method of Establishing Orthotopic Bladder Transplantable Tumor in Mice

Objective The present study aims to find a convenient, rapid, and stable method to establish bladder tumor in mice. Methods Female Balb/C-nu-nu nude mice （or female T739 mice） were narcotized by sodium pentobarbital at a dosage of 60 mg/ kg. The stylet of the 24# venous retention needles was bent in a 5° to 7° angle at a distance of 15 mm from the needlepoint to form a circle with 2.61 mm to 3.66 mm radius when the stylet is rotated. The pipe casing was lubricated with liquid paraffin, and inserted into the bladder cavity. The drift angle stylet was inserted into the pipe casing slowly, rotated for five times, and then pulled out. A cell 6 suspension （0.1 mL） of approximately lx10 T24 cells （or BTT cells） was then injected imme＆ately. Results A total of 60 T739 mice and 60 Balb/C-nu-nu nude mice were inoculated with BTT cells and T24 cells, respectively. The bladder tumor incidence and the average survival time of the tumor-bearing mice were 100% and （26.69±9.24） d and 100% and （34.59±9.8） d for the T739 mice and Balb/C-nu-nu nude mice, respectively. Conclusions Using the drift angle stylet to injure the mucous membrane of the urinary bladder can establish a stable bladder transplantable tumor model in mice.

Inhibitory effect of parvovirus H-1 on the formation of colonies of human hepatoma cell line in vitro and its tumors in nude mice

The inhibitory effect of parvovirus H-1 on the colonyforming ability in vitro of QGY-7703, a cultured human hepatoma cell line, and on the formation and growth of its tumors in nude mice was studied. With higher multiplicity of infection (MOI) of H-1 given, survival of the QGY-7703 cells was found to be decreased. H-1 DNA amplification level at 30 h postinfection(p.i.) was detected to be 7.4 times higher than that at 2 h by dispersed cells assay, while the cells were delayed to enter into S phase.Plaques were formed in the indicator cells (new-born human kidney cell line, NBK) by progeny H-1 virus particles released from the infected QGY-7703 cells by infectious cell center assay. The formation of tumors in nude mice by QGY-7703 cells which were injected s c at 2 h postinfection was observed to be prevented in 2 groups with given MOI 25 and 50. The tumor growth of MOI 10 group occurred at a lower exponential rate than that of control,after a 20 d latent period. It was evident that parvovirus H-1 exhibited a direct inhibitory effect on the formation and growth of human hepatoma cells in vivo as well as in vitro.

Microencapsulated tumor assay:Evaluation of the nude mouse model of pancreatic cancer

AIM: To establish a more stable and accurate nude mouse model of pancreatic cancer using cancer cell microencapsulation. METHODS: The assay is based on microencapsulation technology, wherein human tumor cells are encapsulated in small microcapsules (approximately 420 μm in diameter) constructed of semipermeable membranes. We implemented two kinds of subcutaneous implantation models in nude mice using the injection of single tumor cells and encapsulated pancreatic tumor cells. The size of subcutaneously implanted tumors was observed ona weekly basis using two methods, and growth curves were generated from these data. The growth and metastasis of orthotopically injected single tumor cells and encapsulated pancreatic tumor cells were evaluated at four and eight weeks postimplantation by positron emission tomography-computed tomography scan and necropsy. The pancreatic tumor samples obtained from each method were then sent for pathological examination. We evaluated differences in the rates of tumor incidence and the presence of metastasis and variations in tumor volume and tumor weight in the cancer microcapsules vs single-cell suspensions. RESULTS: Sequential in vitro observations of the microcapsules showed that the cancer cells in microcapsules proliferated well and formed spheroids at days 4 to 6. Further in vitro culture resulted in bursting of the membrane of the microcapsules and cells deviated outward and continued to grow in flasks. The optimum injection time was found to be 5 d after tumor encapsulation. In the subcutaneous implantation model, there were no significant differences in terms of tumor volume between the encapsulated pancreatic tumor cells and cells alone and rate of tumor incidence. There was a significant difference in the rate of successful im- plantation between the cancer cell microencapsulation group and the single tumor-cell suspension group (100% vs 71.43%, respectively, P = 0.0489) in the orthotropic implantation model. The former method displayed an obvious advantage in tumor mass (4th wk: 0.0461 ± 0.0399 vs 0.0313 ± 0.021, t = -0.81, P = 0.4379; 8th wk: 0.1284 ± 0.0284 vs 0.0943 ± 0.0571, t = -2.28, respectively, P = 0.0457) compared with the latter in the orthotopic implantation model. CONCLUSION: Encapsulation of pancreatic tumor cells is a reliable method for establishing a pancreatic tumor animal model.

Establishment of an orthotopic transplantation tumor model of hepatocellular carcinoma in mice

AIM:To improve the outcome of orthotopic transplantation in a mouse model,we used an absorbable gelatin sponge(AGS) in nude mice to establish an orthotopic implantation tumor model.METHODS:MHCC-97L hepatocellular carcinoma(HCC)cells stably expressing the luciferase gene were injected into the subcutaneous region of nude mice.One week later,the ectopic tumors were harvested and transplanted into the left liver lobe of nude mice.The AGS was used to establish the nude mouse orthotopic implantation tumor model.The tumor suppressor gene,paired box gene 5(PAX5),which is a tumor suppressor in HCC,was transfected into HCC cells to validate the model.Tumor growth was measured by bioluminescence imaging technology.Semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) and histopathology were used to confirm the tumorigenicity of the implanted tumor from the MHCC-97L cell line.RESULTS:We successfully developed an orthotopic transplantation tumor model in nude mice with the use of an AGS.The success rate of tumor transplantation was improved from 60% in the control group to 100% in the experimental group using AGS.The detection of fluorescent signals showed that tumors grew in all live nude mice.The mice were divided into 3 groups:AGS-,AGS+/PAX5-and AGS+/PAX5 +.Tumor size was significantly smaller in PAX5 transfected nude mice compared to control mice(P < 0.0001).These fluorescent signal results were consistent with observations made during surgery.Pathologic examination further confirmed that the tissues from the ectopic tumor were HCC.Results from RT-PCR proved that the HCC originated from MHCC-97L cells.CONCLUSION:Using an AGS is a convenient and efficient way of establishing an indirect orthotopic liver transplantation tumor model with a high success rate.

Interleukin-18 suppresses tumor growth and induces tumor cells apoptosis on implanted Lewis lung cancer

Objective： The aim of our study was to investigate the effects of interleukin-18 （IL-18） on implanted Lewis lung cancer in suppressing tumor growth and inducing tumor cells apoptosis. Methods： One week after hypodermic inoculation of Lewis cells, sixteen tumor-bearing syngeneic mice were randomly divided into two groups. The mice in the treatment group were intrapedtoneal injected with the IL-18, in the control group were intrapedtoneal injected with normal saline. The health conditions of the mice were measured, the morphology of the tumors was studied and the apoptosis of implanted lung cancer cells was detected by TUNEL to evaluated the effects of IL-18. Results： IL-18 had no significant effects on the health condi- tions of the mice, but it could suppress the implanted tumor growth （inhibition rate was 75%） and induce tumor cells apoptosis. Conclusion： IL-18 can suppress the implanted Lewis lung cancer cells by way of inducing tumor cells apoptosis. It could be used as a new strategy for lung cancer.