A Study on Liver Damage Induced by Photodynamic Therapy

目的　研究光动力作用对活体肝组织的损伤 ,探讨光动力治疗肝癌的可行性 ,为临床治疗提供实验依据。方法　动物实验 :将小鼠分成光动力疗法 (PDT)组、单血卟啉衍生物 (HpD)组、激光组和空白对照组。光敏药物选用血卟啉衍生物 ,给药量每公斤体重 10mg ,药物用 1ml生理盐水稀释 ,于实验前 48h将药物注射入PDT组和HpD组小鼠腹腔内 ,避光饲养。将PDT组和激光组小鼠固定于实验板上。麻醉后 ,剖腹暴露右肝前叶 ,激光直接投照于肝脏表面 ,光斑直径 5mm ,照射 2min。激光器为氩离子泵浦染料激光器系统 ,光波长 6 30nm ,输出功率 10 0mW ,每一照射区能量累积约 6 0J ,照光后关腹 ,回笼饲养观察。于照光后 1、2 4、72、12 0h处死各组小鼠 ,剖腹取肝组织置于 4%福尔马林液中固定 ,常规石蜡包埋切片 ,HE染色 ,光镜观察。临床治疗 :经病理确诊的肝癌患者 ,于治疗前 48h做皮肤划痕试验 ,阴性者按每公斤体重 5mg静脉给药。治疗时 ,在B超引导下 ,用 18G肝穿针经皮穿刺 ,将石英光纤导入肝肿瘤内。激光波长 6 30nm ,输出功率 35 0mW ,每一照射点能量累积2 2 0J。治疗 1个月后行二期切除术。标本用 4%福尔马林固定 ,常规石蜡包埋切片 ,HE染色 ,光镜观察。结果　动物实验光镜观察结果显示 :PDT组于照光后 2 4h出现照光区肝细Objective To investigate the liver damage induced by photodynamic therapy(PDT) and provide an experimental basis for PDT treatment for liver cancers. Methods 96 normal mice were divided into 4 groups: PDT group, laser group, HpD group and control group. The photosensitizer used in this study was hematoporphyrin derivative (HpD), diluted in 5% glucose and injected into the peritoneal cavity at a dose of 10 mg/kg body weight 48 h before light irradiation. The mice were kept from sunlight exposure. After anesthesia the abdomen was opened and the right front lobe of the liver was exposed. An argon laser pumped dye laser system was used. The liver surface was directly irradiated by the 630 nm laser beam at a power of 100 mW for 2 minutes. The spot size was 5 mm in diameter and the energy density was 60 J/cm 2. The mice were killed at 1, 24, 72 and 120 hours after laser irradiation, respectively. Samples were embedded in paraffin and HE stained sections were examined underlight microscope. Besides, a 46-years old male patient with liver cancer was also included in this study. He received HpD in a dose of 5 mg/kg body weight, i.v. injected 48 h prior to laser irradiation. Ultrasound-guided liver puncture was performed and optical fibers were inserted into and evenly distributed in the tumor. The 630 nm laser irradiation was carried out at a power of 350 mW, energy density of 250 J/cm 2 per each spot. The patient was operated one month later and specimens were taken for histopathological examination. Results Animal experiment: Large necrotic areas were observed in livers of mice 24 hours after PDT. There was a clear demarcation between irradiated and non-irradiated areas observed by both gross and microscopic examination. Fibrous proliferation was seen in the surrounding tissues 120 hours after PDT. Swelling of hepatocytes was observed at 1 h after laser irradiation alone, but returned to normal at 72 h after irradiation. No damage to hepatocytes was observed in livers of both HpD alone and control groups. Clinical case: Wide-spread necrotic areas were present in the PDT irradiated tumor tissue. Normal hepatocytes were observed in the non-irradiated surrounding tissue. There were numerous lymphocytes and macrophages infiltrating in the surrounding areas. Conclusions Selective and sharply demarcated photodamage to liver tissue can be induced by selective laser irradiation after HpD administration. It is suggested that photodamage to surrounding normal tissues can be avoided by carefully controlled laser irradiation during photodynamic therapy of liver cancers.福建省“95”重点科技项

人肝癌裸鼠移植癌株HHC_4、HHC_（15）分子病理学研究

厦门市同安地区已成为中国第二肝癌死亡率高发区。我们对从该地区建立的人肝癌裸鼠移植瘤株，（HHC＿4、HHC＿（15））细胞dnA进行分子生物学研究。应用PCr方法，我们证明了HHC＿4、HHC＿（15）癌细胞dnA都有HbVdnA整合。通过dnA测序，我们证明了HHC＿4dnA有P53基因第250密码子的突变；HHC＿（15）有P53基因第249密码子的突变。这为本地区肝癌死亡率与高HbV高感率和/或黄曲霉素b＿1在食品中高污染率的关系提供有力的分子病理学证据

Establishment of laboratory animal model of cagA(+) coccoid Helicobacter pylori

目的　建立Hp圆球体动物模型。 方法　通过去营养法来培养cagA(+)Hp圆球体 ,定期用MTT和SCC方法来检测细菌的代谢活性 ,用吉姆萨染色和电镜负染色来观察细菌在不同时期的形态和变化。在圆球体培养的第 7周 ,进行幽门螺杆菌及其圆球体经口感染裸鼠的实验。结果　通过细菌学、尿素酶实验、16SRNA、cagA(+)PCR检测和电镜负染色等方法确定从已喂服幽门螺杆菌及其圆球体的裸鼠胃里分离到的细菌克隆 ,经证实为cagA(+)Hp。 结论　首次在裸鼠上证实了Hp圆球体是Hp“粪—口”传播链上不可缺少的一环。Aim To establish animal model infected with coccoid Hp Methods cagA(+) coccoid Hp was cultured and formed through nutrition deprivation,its metabolism was studied regularly with the methods of SCC and MTT the forms of different time was observed by Giemsa staining and TEM Experiment of infecting nude mice was done with cagA(+) coccoid Hp cultured as mentioned above for 7 weeks and with nomal cagA(+)Hp Result Bacteria isolated from nude mice infected by Hp or its coccoid form were comfirmed to contain cagA(+)Hp with assays of PCR,TEM and Urease Conclusion The results indicate that the coccoid form is a necessary link of Hp's "stool-mouth" infection in nude mic

溶癌灵悬乳剂肿瘤内注射的抗癌作用实验研究

目的:探讨溶癌灵合并鸦胆子种皮毒性部分研制新型肿瘤内注射药物的抗癌效果。方法:经乙醇抽提的鸦胆子种皮水溶性部分,以聚乙烯吡咯烷酮为载体,混合消癌灵制成溶癌灵悬乳剂,并以体外细胞毒杀伤及荷瘤动物肿瘤内注射抑瘤实验观察。结果:细胞毒实验显示该剂对体外培养细胞的杀伤效果与鸦胆子种皮提取液相当,但优于消癌灵及5-Fu;体内抑瘤实验表明在抑瘤作用及伤口愈合等方面,该制剂均优于消癌灵、酒精及5-Fu 组。结论:溶癌灵是一种适于肿瘤局部注射治疗的细胞毒性物质,有一定的开发前景

人肝癌裸鼠移植瘤株p53基因突变及测序研究

目的研究从厦门同安肝癌高发区所建的裸鼠移植瘤（ＨＨＣ４、ＨＨＣ１５）是否存在ｐ５３基因突变。方法从两移瘤提取的总ＤＮＡ用分子生物学方法检测并测序，如ＰＣＲ方法扩增ｐ５３基因部分第７外显子，地高辛标记ＤＮＡ探针斑点杂交法，ＤＮＡ限制性酶切片段长度多态分析和激光荧光ＤＮＡ测序等。结果ＨＨＣ４细胞ＤＮＡ有ｐ５３基因第２５０密码子（Ｃ→Ａ）的突变；ＨＨＣ１５有ｐ５３基因第２４９密码子（Ｇ→Ｔ）的突变。结论ＨＨＣ４、ＨＨＣ１５ｐ５３基因突变是肝癌发生的基础，与移植瘤来源地区ＡＦＢ１的严重污染可能有关系

人肝癌裸鼠移植瘤株HHC_4、HHC_(15)的分子病理学研究

厦门市同安地区已成为中国肝癌死亡率第二的高发区。作者对由来自该地区肝癌病人的人肝癌所建立的裸鼠移植瘤株(HHC_4、HHC_(15)细胞所提取的dnA进行分子生物学研究。应用PCr方法发现了HHC_4、HHC_(15)癌细胞dnA都有HbVdnA整合。通过dnA测序,证明了HHC_4dnA有P53基因第250密码子的C→A突变;HHC_(15)有P53基因第249密码子的g→T突变。这为本地区肝癌死亡率与高HbV感染率和/或黄曲霉素b_1在食品中高污染率的关系提供有力的分子病理学证据

Preliminary pharmacodynamic study of STS as an adjuvant ofthe chemotherapy drugs in Vitro and in Vivo

目的：研究硫代硫酸钠（STS）是否能在体内、外作为化疗药的辅药。方法：应用MTT法研究STS分别与AdM、MMC和CddP等6种抗癌单药合用时对bEl-7402和MgC80-3细胞的细胞毒性作用；应用小鼠肝癌腹水瘤（H22）模型判断STS与AdM等3种药合用时的抗癌疗效；20例人原发性肝癌病人肝动脉插管化疗（AdM、MMC和CddP）前30MIn静脉推注STS（125~25g/M2）考察STS作为化疗药辅药的作用。结果：除了CddP以外，STS（500μg/Ml）与AdM等5种抗癌单药合用时对抗癌药的抗肿瘤细胞活性无明显影响（P＞005）。当不同剂量STS（350Mg/kg、35Mg/kg）分别与AdM（6Mg/kg）、MMC（14Mg/kg）和CddP（45Mg/kg）合用或仅这3种化疗药合用时，这3组腹水瘤小鼠的存活期比对照组者显著延长（P＜0001），但3组之间无明显区别（P＞005）。人肝癌肝动脉插管化疗时，STS与AdM、MMC和CddP抗癌药合用，肝癌治疗总有效率达60%，且可减少70%病人的恶心、呕吐等副作用。结论：在体内，低浓度的STS（125~25g/M2）可与AdM，MM？Objective: To study whether sodium thiosulfate (STS) can be used as an adjuvant of the chemotherapy drugs. Methods: The cytotoxicity of STS combined with ADM, MMC, CDDP, 5Fu, MTX and VCR (1PPC/ml) respectively on the cells of BEL7402 and MGc803 was studied by MTT test in vitro.The ascitic hepatoma (H22) in mice were adopted to determine the anticancer effect of STS adjuvant of ADMMMC and CDDP.Being treated with STS(12525 g/m2) 30 min before drug administration,20 patients of hepatocellular carcinoma (HCC) were carried on transcatheter arterial chemoembolization with ADMMMC and CDDP to observe the adjuvant effect of STS. Results: The anticancer activity of ADM and the other agents respectively adjuvant with STS (500 g/ml) were not obviously influented (P >005), but that for CDDP was influented .It was proven that the survival time of ascitic hepatoma treated with ADM (6 mg/kg),MMC (14 mg/kg) and CDDP (45 mg/kg) alone or three drugs in addition to STS (350 mg/kg,35 mg/kg) respectively was not significantly different (P >005).The survival time of each groups was significantly longer than that of control group (P< 0001).It have also been proven that STS adjuvant of ADM, MMC and CDDP was not decreased the chemotherapry effect (RR=60%) by transcatheter arterial chemoembolization in HCC, simultaneously it alleviated the nausea and vomiting in 70%patients. Conclusion: The study provides evidence that in vivo, STS (12525 g/m2) can be used as an adjuvant of chemotherapy drugs,such as ADM,MMC, including CDDP, and so on , for decreasing nausea and vomiting effects, espacially for transcatheter chemotherapy .福建省“八五”肝癌攻关课

李永康膏滋对小鼠胸腺端粒酶活性等免疫功能的影响

通过臭氧(O_3)致小鼠衰老模型,对各组抗衰老实验小鼠的生理和免疫功能进行检测,采用端粒重复序列分析(TRAP)方法检测衰老小鼠胸腺组织的端粒酶活性。李永康膏滋能使衰老小鼠爬绳、游泳和抗冻能力增强(P<0.05或P<0.01),血清总IgG浓度提高(P<0.01),脚掌发疱增重明显(P<0.01),胸腺指数增加(P<0.05或P<0.01);并使衰老小鼠胸腺端粒酶活性增强。表明李氏膏滋有延缓臭氧致小鼠衰老的作用,提高衰老小鼠体液和细胞免疫功能。使衰老小鼠胸腺端粒酶活性增强可能与该器官富含活化的淋巴细胞有关。教育部厦门大学细胞生物学和肿瘤细胞工程重点实验室开放研究基

用荷瘤裸鼠模型研制抗人肝癌单克隆抗体

1985年Watanabe等报道:用去胸腺载瘤小鼠的脾细胞与小鼠骨髓瘤细胞SP 2/0融合获得抗结肠癌单克隆抗体之后,利用荷瘤裸鼠模型成功研制抗人肿瘤单抗的报道已相继出现:但所获单抗多为IgM,IgG类的单抗得率不高,本工作采用多次手术大部切除瘤块的方法延长荷瘤裸鼠的成活期,获得一株抗人肝癌IgG类单抗、其研制过程和结果报道如下

Isoverbascoside对HL-60细胞的诱导分化和细胞毒作用

不同时间、不同浓度的isoverbascoside体外处理HL-60细胞,以形态改变(光镜和透射电镜观察)、功能分化(化学发光检测吞噬能力)、恶性度降低(裸鼠成瘤试验)等指标观察其诱导分化作用;以台盼蓝拒染作用和电镜下形态变化确定其细胞毒作用;用流式细胞术测定其对HL-60细胞周期的影响。20—25μmol/L Isov 1—3天诱导HL-60细胞向粒系方向分化,细胞吞噬能力提高,裸鼠成瘤性降低。30—35μmol/L Isov 2—3天对HL-60细胞有强烈的细胞毒作用。20μmol/L Isov处理12h可引起HL-60细胞的G_1期阻滞,在72h时引起HL-60细胞的G_2/M期的阻滞