DNA-天坛痘苗复合型艾滋病疫苗小鼠体内生物分布研究

目的考察DNA-天坛痘苗复合型艾滋病疫苗在小鼠体内生物分布情况。方法采用定量PCR方法分别对复合疫苗免疫后DNA疫苗的分布情况以及复合疫苗组分HIV-痘病毒载体疫苗单独免疫后的生物分布情况进行测定。结果复合疫苗免疫后5d和30d,主要是在注射部位检测到质粒DNA残留,在免疫后第56d,各脏器均未检测到质粒DNA残留。HIV-痘病毒载体疫苗单独免疫后第5d,仅在给药部位检测到DNA残留,免疫后第36d,各脏器均未检测到DNA残留。结论复合疫苗免疫机体后短期只在注射部位分布,且随时间延长减少至消失,不会长期在机体内存留。     

不同剂量环磷酰胺对小鼠的免疫毒性研究

采用１２,２５,３５ｍｇ／ｋｇｂｗ三个剂量环磷酰胺,经口灌胃给药,连续１８天,测定小鼠的免疫器官的脏体系数,外周血Ｔ淋巴细胞ＡＮＡＥ阳性率,以及碳粒廓清实验的廓清指数。结果表明：ＣＰ在１０ｍｇ／ｋｇｄｗ剂量时,除对ＡＮＡＥ阳性率有明显降低外,对其余两个指标的影响均不明显,而在２５ｍｇ／ｋｇｂｗ,３５ｍｇ／ｋｇｂｗ时,对所试的三个指标有明显降低。     

治疗型双质粒HBV-DNA疫苗长期毒性试验——小鼠肌肉注射给药联合电转染

目的：治疗型双质粒HBV-DNA疫苗是中国人民解放军第458医院肝病研究所研制的生物技术新药。为了观察连续给予治疗型双质粒HBV-DNA疫苗后对小鼠产生的毒性反应及其严重程度，提供毒性反应的靶器官及其损害的可逆性资料，确定无毒反应剂量，给拟定人用安全剂量提供参考，进行长期毒性试验。     

DNA疫苗用于防治艾滋病的研究概况

目的:将能表达抗原的DNA作为疫苗接种。方法:直接注射(皮下,肌肉等)含有DNA的制剂。结果:接种后机体可产生体液免疫和杀伤性T淋巴细胞(CTL)反应,并使机体产生抗病保护。结论:DNA疫苗将有可能用于防治人类艾滋病的理想化疫苗。     

DNA疫苗用于防治艾滋病的研究概况

目的:将能表达抗原的DNA作为疫苗接种。方法:直接注射(皮下,肌肉等)含有DNA的制剂。结果:接种后机体可产生体液免疫和杀伤性T淋巴细胞(CTL)反应,并使机体产生抗病保护。结论:DNA疫苗将有可能用于防治人类艾滋病的理想化疫苗。     

双氢青蒿素对Balb/c小鼠免疫功能的影响

目的 观察双氢青蒿素对Balb/c小鼠非特异以及特异性免疫功能的影响。方法 采用经口方式给药，设31.25、62.5、125和250 mg/kg 4个剂量组，并设溶剂对照组，时间两周。观察双氢青蒿素对淋巴器官脏器湿重以及脏器系数的影响，采用乳酸脱氢酶释放法检测NK细胞活性，流式细胞技术测定巨噬细胞吞噬功能，溶血空斑实验...     

重组痘苗病毒艾滋病疫苗细胞免疫评价方法的优化

目的  优化酶联免疫斑点试验（enzyme-linked immunospot assay,ELISPOT）方法,提高其在重组天坛株痘苗病毒艾滋病疫苗细胞免疫评价中的灵敏度。方法  将疫苗或对照痘苗病毒皮内免疫BALB/c小鼠,50 μl/只,6周后无菌采集小鼠脾脏。用ELISPOT对分泌IFN-γ的脾淋巴细胞〔斑点形成细胞（spot-forming cell,SFC）〕进行检测。比较不同脾淋巴细胞密度（2×10⁵、1×10⁶、2×10⁶个/孔）和不同刺激肽质量浓度（2、4 μg/ml）对SFC的影响。分别采用单因素方差分析和t检验进行多组间和两组间数据比较。结果  3个细胞密度组（F=0.023,P>0.05）和2个刺激肽浓度组（t=-1.762~-0.110,P值均>0.05）间的SFC数差异均无统计学意义。当细胞密度为2×10⁵个/孔、刺激肽质量浓度为4 μg/ml时,疫苗组的SFC数达到（107.00±42.01）个/106细胞,高于原实验条件（细胞密度为1×10⁶个/孔、刺激肽质量浓度为2 μg/ml）的SFC数〔（76.55±64.45）个/10⁶细胞〕,但两者间的差异无统计学意义（t=1.252,P>0.05）。前者的SFC阳性率达到100%,而后者仅为50%。结论  通过优化ELISPOT方法中的细胞密度和刺激肽浓度,进一步提升了该法在重组痘苗病毒艾滋病疫苗细胞免疫评价中的灵敏度。     

恒河猴重复给药腺病毒载体疫苗毒性研究

目的 探讨以5型腺病毒为载体的人用疫苗Ad5 SARS-CoV-2(LW2006)经3次皮下重复给予恒河猴所产生的毒性反应,为其安全性评价实验和临床用药提供基础数据。方法 30例恒河猴,雌雄各半,按体重均衡随机分为对照组(生理盐水)、LW2006低剂量(0.5×10¹¹ VP)组和LW2006高剂量(2.0×10¹¹VP)组。通过皮下注射连续给药3周,每周给药1次,恢复期观察4周。于给药结束、恢复观察结束后,分别剖检18只和12只恒河猴。检测观察指标包括临床观察、体重、血液、生化、尿常规、淋巴细胞亚群、细胞因子、抗核抗体、脏器重量、脏器系数和病理学检查。结果 腺病毒疫苗可引起绝大部分动物给药部位皮肤皮下血管周围局灶性/多灶性炎细胞浸润。恢复期观察结束后,以上变化与给药结束后相比,皮下反应动物比例和程度均明显降低,其余检查脏器包括免疫器官胸腺、脾和淋巴结等均未见明显异常变化。结论 在本实验条件下,以5型腺病毒为载体的人用疫苗Ad5 SARS-CoV-2可引起恒河猴给药部位皮下局部轻度刺激性反应,恢复期可逆,无其他毒理学反应。     

破伤风疫苗在BALB/c与NIH小鼠中的免疫效果比较

目的 对比破伤风疫苗在BALB/c与NIH小鼠中的免疫效果,探讨将BALB/c小鼠用于破伤风疫苗效力实验的可能。方法 将破伤风毒素系列稀释至适当的浓度范围,根据小鼠存活情况摸索合适的毒素浓度,重复实验,测定BALB/c和NIH小鼠的破伤风毒素半数致死量（median lethal dose,LD₅₀）。分别用BALB/c和NIH小鼠的破伤风毒素2 LD₅₀同时攻击BALB/c与NIH小鼠,考察BALB/c和NIH小鼠对破伤风毒素的敏感性。将破伤风疫苗稀释50、100、200倍,分别作为高剂量组、中剂量组、低剂量组免疫BALB/c和NIH小鼠,免疫4周后用50 LD₅₀破伤风毒素攻毒,观察小鼠死亡情况。结果   BALB/c小鼠的破伤风毒素2 LD₅₀为0.16 μg/ml;NIH小鼠的破伤风毒素2 LD₅₀为0.23 μg/ml。用0.16 μg/ml的破伤风毒素分别注射BALB/c和NIH小鼠各3组,每组6只,BALB/c小鼠死亡动物数为3、3、2,NIH小鼠死亡动物数为0、0、0。用0.23 μg/ml的破伤风毒素分别注射BALB/c和NIH小鼠各3组,每组6只,BALB/c小鼠死亡动物数为6、6、6,NIH小鼠死亡动物数为3、2、3。3批破伤风疫苗免疫后的BALB/c与NIH小鼠采用相同攻毒剂量,每组14只,BALB/c小鼠攻毒后,高剂量组存活数为13、14、14,中剂量组存活数为10、10、9,低剂量组存活数为4、3、4;NIH小鼠攻毒后,高剂量组存活数为10、10、10,中剂量组存活数为6、7、6,低剂量组存活数为0、0、1。结论   BALB/c小鼠比NIH小鼠对破伤风毒素具有更高的敏感性,能更好地对破伤风疫苗产生免疫应答,在破伤风疫苗效价测定实验中是一种较为理想的小鼠品系。     

丹参注射液小鼠单次给药毒性试验研究

目的 观察单次静脉和腹腔注射给予丹参注射液后对小鼠产生的毒性。方法 采用经典半数致死量（LD₅₀）法进行小鼠单次给药毒性试验,动物分组后以不同的剂量静脉或腹腔注射给药1次,药后观察小鼠反应,检测体重和死亡率,连续观察14 d后剖检,观察主要脏器的病变。结果 丹参注射液小鼠单次静脉和腹腔注射给药半数致死量分别为：3.2 g·kg^-1和6.3 g·kg^-1。死亡小鼠毒性症状主要表现为俯卧不动、抽搐、呼吸困难,昏迷、尿失禁继而死亡。结论 丹参注射液大剂量腹腔和静脉注射可致小鼠全部死亡,但其半数致死量分别为临床用药剂量的65和129倍,安全范围较大。     

Methamphetamine causes acute toxicity in the retina of Balb/c mice

Abstract

Purpose: As a powerful psychostimulant with high potential for abuse, methamphetamine (Meth) could cause neurological diseases. METH-induced ophthalmic complications are present, but its underlying mechanism has not been completely elucidated, specifically on the retina. This study was to investigate effects of Meth treatment on the retina.

Methods: Balb/c mice were treated with Meth at progressively increasing doses (0–6?mg/kg) intraperitoneally four times per day for five days, mice treated with saline as negative control. Electroretinography (ERG) was used to test the function of retina after Meth treatment. Pathological changes were examined by haematoxylin and eosin staining and terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. Enzyme-linked immunosorbent assay (ELISA) was used to measure the norepinephrine and tumour necrosis factor alpha (TNFα). Real-time PCR and western blot were used to measure expression changes of genes and proteins, respectively.

Results: Our data showed that Meth treatment caused photoreceptor cell death and decreased the thickness of retina. Meth treatment also elevated norepinephrine levels in plasma and increased TNFα in the retina. Moreover, Meth treatment decreased platelet endothelial cell adhesion molecule-1 (PECAM-1) protein expression and increased protein expression of matrix metalloproteinases (MMPs) in the retina.

Conclusions: Our study indicated that short-term intraperitoneal treatment of Meth induced retinal degeneration of Balb/c mice due to a vascular loss of PECAM-1 and an increase of MMPs.     

幼龄大鼠重复ig肠炎宁颗粒1个月毒性试验

目的 观察肠炎宁颗粒对幼龄大鼠可能引起的毒性反应和毒性靶器官,为临床应用提供参考。方法 雌性大鼠与雄鼠交配、受孕并生仔。从出生12 d的仔鼠中,筛选出128只（16窝,8只/窝）随机分入4组,每组每性别16只。仔鼠出生第13天,分别ig给予低、中、高剂量（2、4、8 g/kg）的肠炎宁颗粒,对照组给予去离子水,给药体积为10 mL/kg,连续给药31 d,分别于给药31 d和停药28 d次日剖杀。检测指标包括：一般症状、体质量、摄食量、睁眼、腹部出毛、空中翻正、自主活动、尿液、股骨长度、血液学、血清生化学、T淋巴细胞亚群、肉眼大体和镜下组织病理学检查。结果 与对照组比较,给药0~3 d,肠炎宁颗粒低、中、高剂量组雄鼠和低、高剂量组雌鼠的体质量增长、及高剂量组雌雄大鼠给药第3天体质量均明显降低（P<0.05、0.01）;给药31 d,中、高剂量组雄鼠平均股骨长度偏短（P<0.01）,高剂量组雌鼠红细胞（RBC）、血红蛋白（HGB）、红细胞容积（HCT）轻度降低（P<0.05）,给药组脾脏髓外造血增多;停药28 d后均恢复。未见与药物有关的其他不良反应。结论 本实验条件下,肠炎宁颗粒大剂量长期给药可引起幼龄大鼠出现一些异常反应,但程度轻微,且可恢复。     

大鼠肌注伤寒Vi多糖蛋白结合疫苗的长期毒性

目的观察SD大鼠肌注伤寒Vi多糖蛋白结合疫苗的长期毒性.方法疫苗组大鼠(n=24)免疫3针,每次每只25/μg(以多糖量计),第1剂免疫后2周,第3剂免疫后1,3和5周各处死6只大鼠,测血常规、血清测特异性抗体、血清生化,并进行组织器官病检.结果疫苗组大鼠血液学白细胞分类出现可逆性的变化(P＜0.05,P＜0.01),但无明显毒性,血清特异性抗体阳性大鼠病理组织检查未见免疫毒理学损伤.结论伤寒Vi结合疫苗对大鼠有明显的免疫原性,但无明显毒性和局部刺激性.     

麻醉剂对Balb/c小鼠心功能检测的影响

目的 比较麻醉剂水合氯醛、氯胺酮和戊巴比妥钠埘Balb/c小鼠心功能榆测的影响.方法 将36只雌性Balb/c小鼠(8～9周龄)随机分为4组,分别腹腔注射水合氯醛(A)组(4.5 mg/10g,10只)、氯胺酮(B)组(2 mg/10g,8只)、戊巴比妥钠(C)组(0.5 mg/10g,8只),或等量生理盐水(N)组(0.1 ml/lOg,10只).注射后每隔5 mim行超声心动图查一次,至25 min.动态观察心率(HR)、短轴缩短率(FS)、射血分数(EF)和每分输出量(CO).结果 与N组相比,麻醉组HR、FS、EF及CO均显著降低(P＜0.05),以B组最为显著(P＜0.01).A组与C组相比,各检查时间点心功能指标变异系数(CV)降低,25min时降低＞30%.结论 在麻醉状态下,小鼠心功能降低.水合氯醛较戊巴比妥钠或氯胺酮更易获取稳定的心功能指标.     

Acute and subacute (20-d) oral dose toxicity study of modified fluoroquinolone compound 6C in BALB/c mice

Introduction: Antibiotics are compounds used to treat inflammation; fluoroquinolones are antibiotics used in resistant cases. Objective: The purpose of this study was to investigate acute and subacute toxicity for a new modified flouroquinolone compound (MFC 6C) – a broad spectrum antibiotic which was invented at Faculty of Pharmacy in University of Jordan – using BALB/c mice. Materials and methods: In the pilot study (8 groups; 1 Male:1 Female/group), MFC 6C was administered to these mice at dose levels of 500, 600, 700, 1000, 1200, 1600, 3000 and 5000?mg/kg/d. In the subacute study (5 groups; 5 Males:5 Females/group), MFC 6C was administered for two groups at dose levels of 500, 250?mg/kg/d for 20?d by oral gavage; other groups were control groups. Results: Before the acute study, a pilot study was conducted (on 8 separate days) and no mortality was found even at 5000?mg/kg; therefore, LD₅₀ was found to be >5000?mg/kg and no further acute effects need to be investigated; so MFC 6C is slightly toxic. The biochemical study revealed that, in subacute toxicity study (20 consecutive days), MFC 6C 500?mg/kg caused a decrease in male and female mice blood serum SGOT [p?=?0.0189 (for males), 0.0309 (for females)] and a decrease in male mice blood serum CPK level (p?=?0.023). MFC 6C (250?mg/kg) caused a significant decrease of male mice blood serum sugar (p?=?0.04278) and CPK level (p?=?0.005). The histopathological study revealed that, in subacute toxicity study (20 consecutive days), MFC 6C (500?mg/kg) caused; periportal lymphocytic inflammation (male, 60%; female 40%), lymphoid follicle (female, 60%), neutrophilic aggregation and mitotic activity (female, 40%) in the liver. Moreover, it caused interstitial lymphocytic inflammation (male 60%; female 20%) in the kidney. Other changes like follicular hyperplasia (male 40%) were observed in the spleen. It also caused neutrophilic aggregation (male 40%) in the heart. Also, congestion and macrophages were observed. Changes like lymphocytic infiltration (male 20%; female 20%), congestion (male, 20%) and pleural mesothelial hyperplasia (female, 20%) were found in the lungs. MFC 6C (250?mg/kg), in subacute study, caused; lobular lymphocytic infiltration (male 100%; female 100%), portal lymphocytic inflammation (male 40%; female 40%), granuloma and extramedullary hematopoeisis (male 20%; female 20%), apoptotic bodies and plasma collection in the liver. On the other hand, it caused; reactive lymphoid hyperplasia (male 20%; female 20%) in the spleen. Fibrinous pericarditis (male 40%; female 40%), pericardial mesothelial hyperplasia and degenerated myofibers (male 20%; female 20%) were observed in the heart. Parenchymal lymphocytic infiltration was observed in the lungs (male 40%; female 40%). While no changes occurred in testis at the dose (250?mg/kg). No observed effect level (NOEL) was 125?mg/kg/d for 20-d subacute toxicity study. Conclusion: MFC 6C may suppress the function and\or morphology of the body organs.     

Naloxone blocks anxiolytic-like effects of benzodiazepines in Swiss but not in Balb/c mice

 The ability of naloxone to block the effects of the benzodiazepines chlordiazepoxide and diazepam was evaluated in Swiss and Balb/c mice subjected to the light/dark choice test of anxiety or to a choice paradigm for measuring spontaneous exploratory behaviour. In Swiss mice, naloxone (5 or 10 mg/kg) completely or partially suppressed the anxiolytic-like effects of chlordiazepoxide (5 mg/kg) and diazepam (1 mg/kg) in the light/dark test. Naloxone alone was ineffective. None of these compounds affected locomotion in the free exploratory test. In Balb/c mice, naloxone did not reduce the anxiolytic-like action of benzodiazepines in the light/dark test. Moreover, naloxone did not antagonize the decrease in neophobia observed after anxiolytic treatment in Balb/c mice in the free exploratory paradigm. In this strain, benzodiazepines produced an increase of locomotor activity, whereas naloxone decreased it. The stimulant effects of benzodiazepines on locomotor activity were abolished by naloxone. As naloxone (2 mg/kg) reversed the morphine-induced hyperthermia both in Swiss and in Balb/c mice, differences in possible pharmacokinetic factors between the two strains can be ruled out as an explanation for the failure of naloxone to antagonize anxiolytic-like effects in Balb/c mice. Therefore, the ability of naloxone to reverse anxiolytic effects does not hold for all strains of mice. Received: 19 April 1996 / Final version: 22 November 1996     

Raphanus sativus extract protects against Zearalenone induced reproductive toxicity, oxidative stress and mutagenic alterations in male Balb/c mice

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium in cereals and agricultural products. It has been implicated in several mycotoxicosis in farm animals and in humans. There is unequivocal evidence of reproductive toxicity of ZEN in male mice although the mechanism of action is unknown. Several reports suggest that exposure to ZEN resulted in oxidative stress, genotoxicity and perturbation of reproductive parameters. Therefore, the aim of the current study was to evaluate the protective effects of aqueous extract of Raphanus sativus growing in Tunisia against ZEN-induced reproductive toxicity and oxidative stress. Fifty male Balb/c mice were divided into five groups and treated for 28 days as follows: the control group, olive oil-treated groups, another treated with ZEN (40 mg/kg b.w), the last one treated with R. sativus extract alone (15 mg/kg b.w) and the other with ZEN + R. sativus extract. Testis samples were collected for the epididymal sperm count, testosterone concentration, and MDA level, GPx, CAT and SOD activities. Blood samples were collected for different biochemical analyses. Also, RAPD-PCR method was performed to assess the antigenotoxic effect of the extract in germ cells. The results indicated that ZEN-induced toxicological effects in accordance to those reported in the literature: decreasing in the sperm number, testosterone level and antioxidant enzyme status. The RAPD-PCR analysis revealed an alteration in the DNA bands patterns between control and ZEN-treated mice. The extract alone, rich in many antioxidant compounds, was safe and succeeded in counteracting the oxidative stress and protect against the toxicity resulting from ZEN.     

乳腺癌反义基因治疗动物模型建立的研究

目的建立一种简便、可靠的荷人乳腺癌MCF-7细胞移植瘤裸鼠动物模型,以供survivin反义寡核苷酸对乳癌荷瘤裸鼠的抑瘤作用研究。方法常规培养MCF-7人乳腺癌细胞株细胞,注射于30只雌性裸鼠右前肢腋下皮下,以移植后肿瘤体积生长到直径为0.5cm为成瘤,并将这些实验动物分成3组分别以阳离子脂质体、硫化反义寡核苷酸(ASODN)或无义寡核苷酸在不同时间对荷瘤裸鼠的瘤体及瘤周注射3次。观察肿瘤抑制率、细胞凋亡率及细胞凋亡情况。结果成功建立了荷人乳腺癌MCF-7细胞移植瘤裸鼠动物模型,应用Survivin反义核酸技术实验治疗发现ASODN/Lip组抑瘤率达70.1%,细胞凋亡率为38.6%,与其他两组比较有显著性差异(P<0.05)。结论应用裸鼠前肢腋下皮下注射法建立的荷人乳腺癌MCF-7细胞移植动物模型方法简便,成瘤率高,可用于对人乳腺癌反义基因治疗的动物实验研究。