Hepatocellular carcinoma (HCC) is a highly malignant disease, and its outcome of routine therapies is poor. Comprehensive treatment including gene therapy is an important way to improve patients' prognosis and survival. In this study, we successfully constructed a triple-controlled cancer-selective oncolytic adenovirus, QG511-HA-Melittin, carrying melittin gene, in which the hybrid promoter, hypoxia-response element (HRE)-AFP promoter, was used to control viral E1a expression targeting AFP-positive cancer cells in hypoxia microenviroment, and the E1b-55 kDa gene was deleted in cancer cells with p53-deficiency. The cytological experiments found that the viral replication of QG511-HA-Melittin was increased to 12800-folds in Hep3B cells within 48 h, and 130-folds in SMMC-7721, but the virus did not replicate in L-02 cells. QG511-HA-Melittin had a strong inhibition effect on AFP-positive HCC cell proliferation, such as Hep3B and HepG2, whereas, there was low or no inhibition effect of QG511-HA-Melittin on AFP-negative cancer cells SMMC-7721 and normal cells L-02. In the in vivo experiment, compared with the blank control group, QG511-HA-Melittin can significantly inhibit the growth of HCC xenografts (P<0.05). The survival of mice in QG511-HA-Melittin group was much longer than that of the blank control group. Both in vitro and in vivo experiments manifested that QG511-HA-Melittin exerts an inhibitory effect on HCC cells, which may provide a new strategy for HCC biotherapy.

目的:通过实验探讨斛芪浸膏是否具有减轻电离辐射所致胸腺淋巴细胞氧化损伤的作用.方法:取4～6 周健康雄性昆明小鼠,分离胸腺淋巴细胞作原代培养,并分为对照组、辐照组和辐照加低、中、高剂量斛芪浸膏组,除对照组外,其余各组均采用60CO γ射线5Gy 单次辐照.辐照后30min 内分别用流式细胞仪及超氧化物检测试剂盒检测各组细胞内活性氧荧光强度及超氧化物含量.结果:与辐照组比较,辐照加低、中、高剂量斛芪浸膏各组细胞内活性氧强度及超氧化物水平有不同程度下降,差异有统计学意义.结论:斛芪浸膏能在一定程度上降低电离辐射后早期小鼠胸腺淋巴细胞内活性氧水平,具有减轻电离辐射所致的氧化损伤的作用.

OBJECTIVE:To explore the effects of Huqi Extractum (HQE) on the viability and apoptosis in mouse thymic lymphocytes against 60Co radiation.METHODS:Thymic lymphocytes were isolated from 4 -8 weeks healthy male Kunming mice and primarily cultured. Then they were divided into the control group, the irradiation group, the low dose HQE group, the medium dose HQE group, and the high dose HQE group. Equal volume of serum free RPMI-1640 culture solution was added in the control group and the irradiation group, while equal volume of HQE solution (at the daily dose of 25, 50, and 100 mg/mL) was respectively added in the low, medium, and high dose HQE groups. Except the control group, those in the rest groups were exposed radiation at a single dose of 5 Gy gamma-ray. Changes of the thymic lymphocytes' viability were measured by MTT colorimetric assay at 12, 24, 36, and 48 h after radiation. The early apoptosis rate was detected using flow cytometry (FCM) after 10-h radiation. The apoptosis was detected using agarose gel electrophoresis to observe the DNA injury after 24-h radiation.RESULTS:The viability level decreased more obviously in the irradiation group than in the control group at 24 -48 h after radiation (P < 0.01, P < 0.05). The average viability level was obviously higher in the low, medium, and high dose HQE groups than in the irradiation group (P < 0.05) in a dose dependent manner. The early apoptosis rate was obviously lower in the low, medium, and high dose HQE groups than in the irradiation group, with statistical difference shown in the high dose HQE group (P < 0.01). Typical DNA ladder fragments were found in the electrophoresis in all groups except the control group. But the DNA injury was comparatively milder in the low, medium, and high dose HQE groups, with more obvious effects shown in the high dose HQE group.CONCLUSION:HQE showed protection for the viability of early thymic lymphocytes exposed to the 60CO radiation, and could lower the early apoptosis level.

OBJECTIVE To explore the protective effects of Huqi extractum, a compound Chinese herbal medicine, on salivary glands against radiation in Wistar rats. METHODS One hundred Wistar rats were randomly divided into sham-exposure group, untreated group, and low-, medium- and high-dose Huqi groups. Local irradiation of 60Co gamma-rays with a single dose of 15 Gy was applied to the salivary glands of the Wistar rats except the sham-exposure group. After 3- and 40-day treatment, saliva was collected. Colorimetric method, iodine-amylase colorimetric method and enzyme-linked immunosorbent assay were used to detect concentrations of sodium (Na+), potassium (K+) and secretory immunoglobulin A (sIgA) and activity of salivary amylase. Pathological changes of salivary gland tissues were observed by hematoxylin-eosin straining. RESULTS After 3-day administration, radiation-induced salivary gland injuries were obvious and prevalent in irradiated rats. Comparing with the sham-exposure group, saliva concentration of sIgA and body weight were reduced in other irradiated groups, except those in the high-dose Huqi group, while salivary amylase level was increased. At 3-day phase, pathologic changes of the salivary glands were featured as swelling acinus plasm and vacuolation. At 40-day phase, atrophy of gland cells was dominant. After 40-day administration, there were no significant differences between the high-dose Huqi group and sham-exposure group in sIgA and amylase levels and body weight, and according to the histological examination, no significant difference was revealed under the optical microscope. CONCLUSION Chinese herbal medicine is helpful for the recovery of the salivary glands from the radiation injury, morphologically and functionally in rats.