Background: Cimetidine, an antagonist of histamine type II receptors, has shown protective effects against γ-rays or neutrons. However, there have been no reports on the effects of cimetidine against neutrons combined with γ-rays. This study was carried out to evaluate the protective effects of cimetidine on rats exposed to long-term, low-dose-rate neutron and γ-ray combined irradiation（n-γ LDR）.Methods: Fifty male Sprague-Dawley（SD） rats were randomly divided into 5 groups: the normal control group, radiation model group, 20mg/（kg·d） cimetidine group, 80mg/（kg·d） cimetidine group and 160mg/（kg·d） cimetidine group（10 rats per group）. Except for the normal control group, 40 rats were simultaneously exposed to fission neutrons( 252 Cf, 0.085 m Gy/h) for 22 h every day and γ-rays( 60 Co, 0.097Gy/h) for 1.03 h once every three days, and the cimetidine groups were administered intragastrically with cimetidine at doses of 20, 80 and 160mg/kg each day. Peripheral blood WBC of the rats was counted the day following exposure to γ-rays. The rats were anesthetized and sacrificed on the day following exposure to 252 Cf for 28 days. The spleen, thymus, testicle, liver and intestinal tract indexes were evaluated. The DNA content of bone marrow cells and concanavalin A（Con A）-induced lymphocyte proliferation were measured. The frequency of micronuclei in polychromatic erythrocytes（f MNPCEs）, superoxide dismutase（SOD）, malondialdehyde（MDA）, and glutathione peroxidase（GSH-Px） in the serum and liver tissues were detected.Results: The peripheral blood WBC in the cimetidine groups was increased significantly on the 8th day and the 26 th day compared with those in the radiation model group. The spleen, thymus and testicle indexes of the cimetidine groups were higher than those of the radiation model group. The DNA content of bone marrow cells and lymphocyte proliferation in the cimetidine groups were increased significantly, and fMNPCE was reduced 1.41-1.77 fold in cimetidine treated groups. The activities of SOD and GSH-Px in the cimetidine groups were increased significantly, and the content of MDA in the cimetidine groups was decreased significantly.Conclusions: The results suggested that cimetidine alleviated damage induced by long-term, low-dose-rate neutron and γ combined irradiation via antioxidation and immunomodulation. Cimetidine might be useful as a potent radioprotector for radiotherapy patients as well as for occupational exposure workers.

目的 研究海洋星虫多糖原料药的稳定性.方法 海洋星虫多糖原料药经强光照射(4500lx) 10d、高温存放(60℃)10 d、高湿存放[25℃,RH(75±5)％]10 d、加速试验[(40±2)℃,RH(75±5)％]6个月、长期试验1年,考察海洋星虫多糖原料药经不同条件存放后的性状、干燥失重、微生物限度及多糖含量的变化.结果 海洋星虫多糖原料药除高温试验外,其它试验所考察指标均符合质量标准要求.结论 海洋星虫多糖原料药稳定性较好,但具有吸湿性.

Objective To investigate the effects of low dose γ-ray on the gene expression of cyclin G1 (CCNG1) in human lymphoblasts AHH-1 line and the probability of CCNG1 as bio-dosimeter for low-dose radiation. Methods Lymphoblasts of AHH-1 line were irradiated with γ-ray in the doses of 0, 0.1, 0.2, 0.5, 0.8 or 1.0 Gy. Total RNA of cells was extracted at 0, 4, 24, 48, 72 and 168h after irradiation, and real-time Q-PCR was used to detect the expression of CCNG1 gene in AHH-1 cells. The relationship between the irradiation dosage or duration and the gene expression was analyzed. Results The gene expression of CCNG1 in AHH-1 cells increased in a dose-dependent manner after irradiation, presenting a better dose-effect relationship. Time-effect curve indicated that the peak expression of CCNG1 appeared at 24h after irradiation, and then the expression decreased gradually with time, and it recovered to the level before irradiation at 168h after irradiation. Conclusions CCNG1 gene is sensitive to low dose irradiation (0-1.0Gy) with a better dose and time-effect relationship, and it may be used as a potential efficient biological dosimetry in low dose exposure risk assessment. DOI: 10.11855/j.issn.0577-7402.2015.06.16

Objective To optimize the extraction technology of polysaccharide from Sipunculusnudus Linnaeu. Methods The extraction technology of the sipunculusnudus polysaccharide was optimized by the orthogonal design with the solid-liquid ratio,sodium hydroxide concentration,extraction temperature and extraction time as the investigation factors and the product of the polysaccharide content and the polysaccharide yield as the evaluation index. Results The optimal extraction technology of the sipunculusnudus polysaccharide was as follows:the solid-liquid ratio was 1 :4,the concentration of sodium hydroxide was 1%,the extraction temperature was 60 ℃ and the extraction time was 4 h. Conclusion This extraction technology is simple and rapid,which can be applicable for the extraction of the sipunculusnudus polysaccharide.

为观察低剂量γ-射线对人淋巴母细胞AHH1 B细胞转位基因2(B cell translocation gene 2,BTG2)基因mRNA表达水平的影响,探讨BTG2基因作为低剂量范围辐射生物剂量计的可能性,采用0-1.0 Gyγ-射线照射人淋巴母细胞,照射后不同时间点(0-72 h)提取细胞总RNA,采用实时荧光定量PCR方法检测BTG2基因的表达变化,分析该基因的时间-剂量效应关系.结果表明,照射后BTG2基因转录水平表达在低剂量范围呈剂量依赖增加,照射后24 h达峰值,之后逐渐下降,在48 h和72 h时处于平台期,在168 h恢复至初始水平.BTG2基因对低剂量电离辐射敏感,有较好的剂量效应和时间效应关系,具有开发为低剂量辐射生物剂量计的潜力.

Objective To investigate the stability of Sipunculus nudus Polysaccharide Capsules. Methods According to the stability testing of pharmaceutical preparations and guiding principles in Appendix of Chinese Pharmacopeia (2005 edition, 2 volume), with the major investigation items of character, identification, moisture, disintegration time, micro-bial limit test, the content of polysaccharide as the evaluation indexes, the accelerated test and long-term test were performed. Results All indexes in 3 batches of samples after 6-month accelerated testing and 12-month long trial were in line with the quality standard requirements. Conclusion The drug properties of Sipunculus nudus Polysaccharide Capsules are stable.

目的 研究方格星虫多糖(SNPS)对低剂量中子辐射损伤大鼠的保护作用.方法 SNPS 3个剂量组70,140和280mg· kg-1.SD大鼠,50只,雄性,清洁级,180～200 g,每天于照射前经口给予受试药,连续38 d.除正常对照组外,其余各组每天照射22 h,放射源为252 Cf中子源,吸收剂量率为0.085mGy·h-1,累积吸收剂量为71.06 mGy.在照射第7、14、21、28、35 d检测外周血细胞计数,在照射38 d检测血清SOD和GSH-PX活力、MDA含量,血清胆固醇和甘油三酯含量,肝脏脂代谢相关基因CYP7A1、FABP1、PPARα mR-NA表达变化,计算主要脏器指数.结果 与模型对照组比较,在照射第35 d,SNPS中剂量组外周血血小板显著回升,在照射第38 d,SNPS各剂量组血清GSH-PX活力显著上升,血清MDA含量显著下降,肝脏中胆固醇7-α羟化酶含量显著回升.结论 SNPS对长期低剂量中子辐射损伤大鼠具有升高血小板,提高抗氧化能力,调节胆固醇代谢等保护作用.

目的 探讨海洋星虫多糖胶囊的制备工艺条件,并对其理化性质和质量控制进行研究.方法 通过筛选合适的辅料,采用湿法制粒,以颗粒得率、休止角、吸湿性为主要评价指标,以多糖含量作为其质量控制指标.结果 采用微晶纤维素作为填充辅料,具有良好的流动性和崩解性,采用80％乙醇作为润湿剂可克服制粒粘性大,难过筛等情况,所制备颗粒流动性能满足生产需要,其多糖含量为标示量的104.7％.结论 本研究所制备的胶囊颗粒成型性和流动性好,制备工艺稳定,质量可控,适合于工业化生产,可为海洋星虫多糖胶囊的新药申报提供依据.

目的 考察硼卡钠(sodium borocaptate,BSH)对急性辐射损伤小鼠的防护作用.方法 以BSH为受试物,用60Co γ射线对小鼠进行一次性全身照射,照射剂量为6 Gy,剂量率为0.8 Gy/min.照射后第3天、第14天检测外周血细胞计数,并于照射后第14天检测小鼠脏器指数.结果 照后第3天,BSH高剂量组血小板计数显著增加(P＜0.05);照后第14天,与模型组比较,BSH中剂量组血小板计数显著增加(P＜0.01),胸腺指数、脾脏指数均显著升高(P＜0.05).结论 BSH能显著提高辐照后小鼠的血小板水平,对急性辐射损伤小鼠免疫系统有保护作用.

Objective To explore the effect of low-dose γ-rays on the metabolites in rat serum. Methods Sixteen healthy male SD rats were randomly divided into control group and irradiated group (n=8). The rats in irradiated group were irradiated by 60Co γ-rays with a dose rate of 72mGy/h for 7 days (1 hour per day). At the 7th day after irradiation, blood samples were taken from abdominal aorta to obtain the serum. The metabolic fingerprints of serum were obtained from the two groups of rats, and 1H nuclear magnetic resonance (NMR) spectroscopy, principal component analysis (PCA) and orthogonal signal correction-partial least squares (OSC-PLS) method were used for pattern recognition, and the difference in metabolite profile between two groups was identified by SIMCA-P software. Results The rat serum 1H NMR spectra revealed a significant difference between control group and irradiated group, the OSC-PLS plots of the serum samples presented marked clustering between two groups. Compared with the control group, the content of lipid, glucose, creatine, glycine/glucose, high density lipoprotein, low density lipoprotein, very low density lipoprotein/low density lipoprotein and unsaturated fatty acid increased, while the content of lactic acid, threonine/lipid, alanine, N-acetyl glycoprotein 1, N-acetyl glycoprotein 2, saturated fatty acid and phosphatidyl choline decreased in irradiated group. Conclusion Irradiation with low-dose γ-ray could induce changes in metabolites in rat serum, concerning mainly immune function, energy metabolism, carbohydrate metabolism and lipid metabolism. DOI: 10.11855/j.issn.0577-7402.2014.07.02

Objective To investigate the combined biological effect of γ- ray of 60Co,spalling neutron of 252Cf,and noise on rats,so as to provide theoretical basis for explaining the effect of combined environmental factors on the health of special operation population and for working out intervention measures. Methods Sixteen SD rats were randomly divided into experimental group and normal control group. The animals in experimental group were exposed to60Co γ- ray( 0. 5 Gy),noise〔( 90 ± 5) dBA〕,and spalling neutron of252Cf( 12 mGy). Peripheral blood,organ index,marrow DNA content,SOD activity,and MDA content were detected. Results: The level of platelet,spleen index,and thymus index in experimental group〔( 728 × 109) /L,0. 002,and 0. 0017 respectively〕 were significantly lower than those in control group〔( 838 × 109) /L,0. 003,and 0. 0022 respectively〕,( P 0. 05). The marrow DNA content in experimental group( A260: 0. 8862) was also significantly lower than that in control group( A260: 1. 3039),( P 0. 01). Conclusion The hematopoietic system and immune system of rats were markedly damaged by the combined effect of low dose γ- ray,spalling neutron,and noise.

Objective To evaluate the protective effects of sipunculus nudus polysaccharides( SNPS) on mice with irradiation damage induced by 5. 0 Gy60Co γray. Methods 60 mice were divided into control group,model group,nilestriol group and 100 mg / kg·d,200 mg / kg·d,and 400 mg / kg·d SNPS groups. SNPS was administrated by intragastric infusion every day,and nilestriol was administrated by intragastric infusion with 3. 3 mg / kg,1. 67 mg / kg in the day before and after iradiation,respectively. In the fifteenth day,the mice except control group were irradiated by γrays of60Co( 0. 80 Gy / h) with the dose of 5. 0 Gy. Peripheral blood WBC,the spleen index( SI),thymus index( TI),DNA contain in spleen cells and in bone marrow cells,SOD and MDA in serum were detected. Results Peripheral blood WBC in 100mg / kg·d and 200 mg / kg·d SNPS groups were higher than that of model group at the third day after irradiation( P 0. 05). Peripheral blood WBC in 100mg / kg·d,SI in 200 mg / kg·d and 400 mg / kg·d SNPS groups,DNA contain in spleen cells in 200 mg / kg·d SNPS group,DNA contain in in bone marrow cells in 100 mg / kg·d and 200 mg / kg·d SNPS groups,and SOD in serum in three SNPS groups were much higher than that of model group. MDA in three SNPS groups were much lower than that of model group at the fourteenth and third day after irradiation. Conclusion SNPS has radiation protective effects on γrays irradiation mice.

Objective To study the effects of low dose rate 60Co γ ray combined with 252Cf neutron irradiation on hematopoietic and antioxidation systems in rats.Methods Sixteen SD male rats were randomly divided into the irradiation group and the control group,each consisting of 8 rats.The irradiation group was exposed to whole-body 60 Co γray (with daily dose rate of 0.072 Gy) for 1 hour and to 252 Cf neutron (with daily dose rate of 0.085 mGy/h) for 20 hours,for a total of 7 successive days.Peripheral blood cell counts,main organ indices,bone marrow DNA contents,SOD activity and MDA content of rats were monitored,at day 8 after irradiation.Results Compared with those of the control group [(6.2 ± 2.4) × 109/L],WBC of the irradiation group decreased significantly following irradiation [(3.4 ± 1.2) × 109/L],(P ＜ O.05),but no statistical significance could be noted in the changes of various organ indices (P ＞ O.05).Bone marrow DNA contents and serum SOD activity all decreased significantly (P ＜ 0.05),while organ indices and MDA contents seemed to increase,but without statistical significance (P ＞ 0.05).Conclusions 60 Co γirradiation combined with 252Cf neutron irradiation could obviously damage the hemopoietic and anti-oxidation systems of rats.

目的 建立方格星虫多糖的分子量测定分析方法.方法 应用高效凝胶渗透色谱法,色谱柱为phenomenex PolySep-GFC-P 4000(300×7.8 mm),流动相为0.71％硫酸钠溶液(内含0.02％叠氮钠),柱温:35℃,RID-10A示差折光检测器(检测器温度35℃),流速0.5 ml·min-1.结果 根据建立的方法测定,得到方格星虫多糖的高效凝胶色谱图,并计算出其重均分子量.结论 所用方法简便、快速、准确,可用于方格星虫多糖的质量控制.

Objective To study the antihyperlipermia effect of Engraulis Japonicus enzymatic hydrolysate(EJEH) in SD rats.Methods EJEH was prepared from Engraulis Japonicus by enzymatic hydrolysis with 0.05% alkaline proteinase and 0.05% trypsin for 10 hours,and 0.1% Flavored Proteinase for 2 h.The antihyperlipermia effect of EJEH was studied in SD rats.The EJEH were given daily for 35 days,and the model group and the control group were given with distilled water.At the 35th day,serum TC,TG,HDL-C,LDL-C,LDL-C/HDL-C,TG/HDL-C,SOD,MDA,liver weight,and microscope pathomorphism were tested.Results The 1evel of the serum TC in the control group was 1.359±0.147 mmol·L-1,in the model group was 1.590±0.056 mmol·L-1(P0.01,comparing with the control group),in the 70 mg/kg d-1 Xuezhikang positive control group was 1.485±0.102 mmol·L-1(P0.05,comparing with the model group),in the 2.3 g/kg·d-1EJEH group was 1.477±0.098 mmol·L-1(P0.05),and in the 6.9 g/kg·d-1EJEH group was 1.454±0.052 mmol·L-1(P0.01).The level of serum TG in the model group was 1.480±0.260 mmol·L-1,which was increased significantly comparing with the control group(P0.05),and in the three EJEH treated groups the serum TG were decreased significantly comparing with the model group(P0.05,P0.01).The level of serum LDL-C in the model group was 0.960±0.090,which was increased significantly comparing with the control group(P0.05),and in the 6.9 g/kg·d-1EJEH group was 0.849±0.054 mmol·L-1(P0.05,comparing with the model group).The results suggested that EJEH decreased the serum levels of TC,TG,and LDL-C significantly.In the 6.9g/kg·d-1EJEH group,the serum SOD activity was increased significantly comparing with the model group(P0.05).The liver adipose degeneration in the Xuezhikang positive control group and the EJEH treated groups were ameliorated notably.Conclusions EJEH has significant Antihyperlipermia Effect in SD Rats.

目的观察方格星虫多糖对亚致死剂量60Coγ射线照射小鼠的保护作用。方法将60只雄性C57BL/6J小鼠随机分为正常对照组、模型组、尼尔雌醇组及低（100 mg/kg·d）、中（200 mg/kg·d）、高（400 mg/kg·d）三个剂量方格星虫多糖组,每天一次灌胃给药,尼尔雌醇组照射前一天给予尼尔雌醇3.3 mg/kg灌胃,照射后0.5 h再次给予1.67 mg/kg尼尔雌醇灌胃。灌胃2 w后,除正常对照组外,各组均以剂量率为0.8 Gy/min的60Coγ射线全身照射一次,照射剂量为5.0 Gy。于辐照后3 d、14 d测定小鼠体重、外周血WBC,辐照后14 d测定小鼠胸腺指数,脾指数,骨髓细胞和脾细胞DNA含量,血清SOD和MDA含量。结果γ射线全身照射后第3天,方格星虫多糖低、中剂量组小鼠外周血WBC数明显升高（P〈0.05）。照后第14天,低剂量组小鼠外周血WBC数显著增高（P〈0.01）,中、高剂量组脾脏重量指数明显升高（P〈0.01和P〈0.05）,中剂量组脾脏DNA含量明显增高（P〈0.01）;低、中剂量组骨髓细胞DNA含量明显增高（P〈0.05）;低、中、高三个剂量组小鼠血清SOD活力明显增强,血清MDA含量明显下降（P〈0.05）。结论方格星虫多糖对亚致死剂量γ辐射损伤小鼠具有一定保护作用。

To investigate the mechanism of biological effects induced by low dose radiation,the alterations of transcriptional profiles of human normal lymphoblastoid cell AHH-1 lines irradiated with different doses of γ-rays were assayed.AHH-1 lines were irradiated with γ-rays of 0.1,0.5,and 1.0 Gy,respectively,with the unirradiated cells as control group.Total RNA of irradiated cells was extracted 4 h after irradiation,and then cDNA microarray was used to detect the transcriptional profiles of the cells.The different expressions of genes were assayed by hierarchical cluster analysis,gene ontology and pathway analysis,and then identified by real-time Q-PCR.The results of real-time Q-PCR of CASP9 were in accordance with microarray assays.The microarray assays showed that there were 1 330 up-regulated genes and 1 002 down-regulated genes in three irradiated groups.Alterations of 18 genes expression were related with absorbed dose.Among these genes,16 genes were positive relation,and 2 genes were negative relation with dose.The results of hierarchial cluster analysis indicated that the similarity of the expression profiles of 0.1 Gy and 1.0 Gy was the highest among 4 groups.These altered genes could be related to multiple signal transductions,such as cell cycle,p53 signal pathway,base excision repair,RNA transport,endoplasmic reticulum proteins processing pathways,and so on.These differential expressed genes detected by cDNA microarray would help to elucidate the mechanisms of biological effects of low dose irradiation.New biological dosimeter for low dose exposure might be developed from these dose-related genes.

Objective: To establish a simple,rapid and accurate method of protein content analysis of polysaccharide from Sipunculus nudus. Method: Using Kjeldah method and Coomassie brilliant blue method to test protein content of the polysaccharide from S. nudus. Also the precision and accuracy of the two methods were tested. Result: The content of protein was 2. 40% and 2. 22% by the two method and the average recovery rates were 98. 69% and 100. 97% respectively. The content by the two methods was almost the same and had no marked difference( P 0. 05). Conclusion: In summary,Kjeldah method is simple and accurate and could assay the protein content of polysaccharide from S. nudus rapidly.

Objective To investigate the protective effect of Acanthopanax Senticosus polysaccharide on peripheral hematological cells in rats irradiated with 60Co γ ray.Methods Sixty-three male rats were randomly divided into 7 groups,i.e.the normal control group,the irradiation control group,the WR-2721 group,the 523 group and the 3 experimental groups with high (200 mg · kg-1 · d-1 ), moderate (100 mg · kg-1 · d-1) and low (50 mg· kg-1 · d-1) doses of Acanthopanax Senticosus polysaccharide.The marrow-depressed model rats received 5 Gy 60Co γ ray whole body irradiation with a dose absorption rate of (0.388 Gy/h) except those in the normal control group.WBC,RBC and PLT in the peripheral blood were detected with hemocytometer both on the 5th day before irradiation and the 3rd and 10th days after exposure.Results When detected on the 3rd day following irradiation,WBC in the peripheral blood of the irradiated animals decreased to a level of (2.482 ±0.808) × 109 · ml-1,RBC to a level of (7.680 ±0.926) × 1012·ml-1 and PLT to a point of (360.000±43.967) × 109 · ml-1,having a rather significant decrease in the indices,when compared with those of the normal control group (P ＜ 0.01 ). On the 10th day following irradiation,WBC in the peripheral blood of the irradiated control animals decreased to a level of (4.764 ±0.679) × 109 · ml - 1,RBC decreased to (9.004 ± 0.761 ) × 1012 · ml -1 and PLT to ( 390.000 ± 43.967 ) ×109 · ml-1,with all the indices decreasing more obviously than those of the normal control group(P ＜ 0.01 ).Data detection before irradiation with 60Co γ ray indicated that WBC,RBC and PLT in the peripheral blood of the experimental animals elevated to various extent,when compared with those of the normal control group.Detection after irradiation showed that Acanthopanax Senticosus polysaccharide could obviously increase the levels of WBC,RBC and PLT in the peripheral blood of the irradiated animals,when compared with those of the irradiation group. Conclusions Our experimental results showed that Acanthopanax Senticosus polysaccharide seemed to have an obvious protective effect on peripheral hematological cells in marrowdepressed rats irradiated with 60Co γ ray.

Objective To determine the uronic acid in polysaccharides from Sipunculus nudus Linnaeu.Methods The contents of uronic acid in polysaccharide was studied by carbazole and sulphuric acid method with galacturonic acid as standard,the absorbency on 525 nm was measured.Results The linear range of galacturonic acid was 2.75~27.5μg/ml(R2=0.9995).The repitition RSD of uronic acid was 1.22%(n=6) and the average recovery was 103.5%(RSD=1.9%).The contents of uronic acid in polysaccarides of three batchs were 23.68%,22.87% and 25.05%,respectively.Conclusion The established method is simple,sensitive and accurate which can be applied to control the content of uronic acid in polysaccarides from Sipunculus nudus Linnaeu.