补中益气汤对A549/DDP动物模型实体瘤中耐药相关酶GST和TopoⅡ表达的影响

目的研究补中益气汤对A549/DDP细胞荷瘤小鼠实体瘤中耐药相关酶GST和TopoⅡ表达的影响,探讨补中益气汤的作用机制。方法体外培养A549/DDP细胞,接种健康BALB/c小鼠,荷瘤成功后予补中益气汤治疗,实验分为:荷瘤组,补中益气汤低剂量组及补中益气汤高剂量组。取实体瘤后,利用RT-PCR检测GST和TopoⅡmRNA的表达,免疫组化法检测蛋白表达。结果同荷瘤组相比,高低剂量的补中益气汤均可降低耐药相关酶GST和TopoⅡmRNA表达水平,但补中益气汤高剂量组效果同低剂量组区别不大(0.05)。同荷瘤组相比,高低剂量的补中益气汤均可降低耐药相关酶GST和TopoⅡ蛋白的表达(0.05),补中益气汤高剂量组的效果优于补中益气汤低剂量组(0.05)。结论补中益气汤能下调A549/DDP实体瘤中耐药相关酶GST和TopoⅡ的表达水平。     

顺铂联合雷帕霉素、3-MA对裸鼠肺腺癌A549细胞原位移植瘤的影响

目的探讨顺铂(DDP)分别联合雷帕霉素(Rapamycin,RAPA)、3-甲基腺嘌呤(3-Methyladenine,3-MA)对裸鼠肺腺癌A549细胞原位移植瘤的生长抑制作用和相关机制。方法采用RT-PCR、免疫组化SP法检测经药物作用后裸鼠移植瘤中m TOR、LC3-Ⅱ及Bax基因和蛋白水平的表达。结果 m TOR mRNA在DDP+RAPA组、DDP+3-MA组的表达量显著低于空白组和DDP组(P0.05);LC3-ⅡmRNA在DDP+RAPA组表达量显著高于其它组(P0.05);Bax mRNA的表达量在DDP+3-MA组的相对表达量显著高于其它组(P0.05)。m TOR蛋白在DDP+RAPA组、DDP+3-MA组的表达强度显著低于空白组和DDP组(P0.05);LC3-Ⅱ蛋白在DDP+RAPA组的表达显著高于其它组(P0.05);Bax蛋白在DDP+3-MA组的表达强度显著高于其它组(P0.05)。治疗后空白组、DDP组、DDP+RAPA组及DDP+3-MA组对裸鼠肿瘤的抑制率分别为0、36.5%、43.9%、55.0%。结论 DDP联合RAPA或3-MA均可以明显抑制肺腺癌A549细胞裸鼠移植瘤的生长,并且抑制作用均大于单独使用DDP。     

补中益气汤对A549/DDP肺癌荷瘤BALB/c小鼠耐药相关蛋白MRP表达的影响

目的研究补中益气汤对A549/DDP肺癌荷瘤BALB/c小鼠实体瘤中耐药相关蛋白(multidrug resistance-associated protein,MRP)表达的影响,探讨补中益气汤对A549/DDP的治疗作用机制。方法体外培养A549/DDP细胞,接种健康BALB/c小鼠,荷瘤成功后予补中益气汤治疗,实验分组为:荷瘤对照组,补中益气汤低剂量组及补中益气汤高剂量组,利用RT-PCR检测实体瘤MRP mRNA的表达,免疫组化法检测MRP蛋白表达。结果同荷瘤对照组相比,高、低剂量的补中益气汤均可降低耐药相关蛋白MRP的表达水平,且补中益气汤高剂量组效果同低剂量组相比无明显区别。结论补中益气汤的抗肿瘤作用可能与其降低MRP的表达相关。     

黄芪多糖(APS)及联合顺铂(DDP)抑制小鼠Lewis肺癌复发瘤生长的机制

目的观察黄芪多糖(APS)及联合顺铂(DDP)对小鼠Lewis肺癌术后复发瘤病理形态及转移相关蛋白CD44、CD62P和骨桥蛋白(OPN)表达的影响。方法选取C57BL/6J小鼠10只作为供瘤组,另外80只随机分为模型组、(50、100、200)μg/mL APS处理组、6 mg/kg DDP处理组、3 mg/kg DDP联合(50、100、200)μg/m L APS处理组,每组10只。取供瘤组小鼠瘤组织制备Lewis瘤单细胞悬液接种于每只小鼠左侧后肢爪垫内侧皮下10 d后,切除爪垫瘤组织,建立肺癌术后复发转移模型;造模次日起给药,第16天脱颈处死。采用HE染色法观察术后复发瘤组织病理形态,免疫组织化学染色法检测复发瘤中CD44、CD62P和OPN的表达。结果与模型组相比,各处理组复发瘤组织坏死加重,尤以DDP联合200μg/mL APS处理组更明显,且各处理组CD44、CD62P和OPN蛋白的表达均有降低,以DDP以及DDP联合(100、200)μg/m L APS处理组最为显著。结论DDP联合APS能抑制Lewis肺癌细胞生长并降低瘤组织CD44、CD62P和OPN蛋白表达。     

黄芪多糖联合顺铂对小鼠Lewis肺癌细胞肺转移的影响

目的观察黄芪多糖(APS)联合顺铂(DDP)对小鼠Lewis肺癌细胞肺转移、核因子(NF)-κB、P38、P53及Caspase-9表达的影响。方法将90只Lewis小鼠随机分为模型组、顺铂组(6mg/kg DDP)、APS组(50、100、200)mg/kg,联合用药组[1/2 DDP+APS,即:(3+25、3+50、3+100)mg/kg]。各组均于造模第2天起用药,APS每日1次,DDP每周1次,连续20d。观察肿瘤肺转移情况,采用Real-time PCR法和Western blotting法检测肿瘤组织中NF-κB、P38、P65蛋白和基因,并用免疫组织化学检测Caspase-9的表达。结果与模型组相比,各治疗组均可降低肺转移灶数目(P0.05或P0.01);除P38外,APS中、高剂量组可使小鼠Lewis肺癌组织中NF-κB p65、P53表达降低,Caspase-9表达增高;联合用药高剂量组作用则接近DDP组。结论 APS可抑制小鼠Lewis肺癌细胞的转移,抑制NF-κB和丝裂原活化蛋白激酶(MAPK)信号通路的激活,这可能是其抑制肿瘤转移的机制之一;APS与减半剂量的DDP铂类化疗药物联合使用时,作用增强,APS对DDP有增效减毒作用。     

黄芪多糖联合顺铂对小鼠Lewis肺癌移植瘤Caspase-3、Smac/Diablo表达的影响

目的观察黄芪多糖(APS)及联合顺铂(DDP)对小鼠Lewis肺癌(LLC)移植瘤凋亡蛋白Caspase-3、Smac/Diablo表达的影响,探讨黄芪多糖抗肿瘤的机制。方法 90只C57BL/6 J小鼠,随机分为9组,正常组、模型组、APS低、中、高剂量组、DDP组、联合低、中、高剂量组,每组10只。除正常组外,其余80只均接种肺癌移植瘤细胞(1×1010/L)于右前肢腋窝皮下,制造模型为荷瘤小鼠。制造模型次日起,治疗组的小鼠给予腹腔注射0.3 ml药物。顺铂每周注射1次,其余药物每日1次,正常组和模型组注射等体积生理盐水,连续20 d,于第21天处死。肿瘤组织进行HE染色并行病理学观察;免疫组织化学染色和图像分析方法检测移植瘤细胞中的Caspase-3及Smac/Diablo的表达。结果荷瘤小鼠在APS(高)、联合(中)、联合(高)组的体质量变化,具有统计学意义。与模型组小鼠比较,肿瘤组织的病理组织学HE显示,APS(高)联合顺铂组的肿瘤细胞坏死最为明显;免疫组织化学法表明,治疗组的肿瘤组织中Caspase-3、Smac/Diablo蛋白表达水平均升高,联合(高)组升高最明显。结论 APS及联合化疗药物DDP能抑制小鼠Lewis肺癌细胞的生长,其机制可能与升高Caspase-3、Smac/Diablo的表达有关。     

Flt3L及CCL5对prime/boost免疫策略中抗原特异性免疫应答的增强及抗肿瘤作用

目的:研究Flt3L与CCL5作为联合佐剂在prime/boost免疫策略中对HBc抗原特异性免疫应答的增强及抗肿瘤作用.方法:将两种细胞因子质粒与携带HBc抗原的DNA疫苗经肌内注射法共免疫小鼠, 免疫3次后再用原核表达的HBc颗粒蛋白或HBc DNA疫苗加强, 观察对稳定表达HBcAg 的小鼠黑色素瘤细胞(B16-HBc)的生长抑制作用;并分别采用MTT法检测荷瘤小鼠脾淋巴细胞增殖、流式细胞术检测脾CD8 T淋巴细胞中IFN-γ表达、ELISA法检测脾淋巴细胞培养上清IL-2、IL-4含量及乳酸脱氢酶(LDH)释放法检测特异性CTL杀伤活性.结果:与对照组相比, 佐剂联合DNA疫苗免疫经蛋白加强组(DDP/Adj)显著抑制肿瘤生长;佐剂联合DNA疫苗免疫组(DDD/Adj)及DDP/Adj组均可促进特异性淋巴细胞增殖反应(P＜0.05), 且DDP/Adj 高于DDD/Adj组(P＜0.05);DDD/Adj 及DDP/Adj组小鼠脾脏CD8 T淋巴细胞中IFN-γ表达、IL-2 表达水平及CTL杀靶活性均高于对照组(P＜0.01或P＜0.05), IL-4 表达水平在各组无显著区别(P＞0.05).结论:在prime/boost免疫策略中, 采用Flt3L与CCL5两种细胞因子联合应用可显著促进荷瘤小鼠产生抗原特异性免疫应答及抗肿瘤作用.     

氯喹对S_(180)荷瘤小鼠的抑瘤作用及其机制

目的探讨氯喹(CQ)对小鼠移植性肉瘤S_(180)的抑瘤作用及其可能机制。方法采用40只S_(180)荷瘤小鼠分为模型组,氯喹低剂量组,氯喹中剂量组,氯喹高剂量组进行体内抑瘤实验,观察不同浓度氯喹对小鼠肉瘤S_(180)的抑制作用;透射电子显微镜观察氯喹作用下荷瘤鼠肿瘤细胞超微结构的变化;流式细胞术检测氯喹诱导荷瘤鼠S_(180)细胞凋亡的情况;免疫组织化学方法检测相关凋亡因子Bcl-2、细胞色素C和Cle-Caspase-3的表达情况;Western blotting检测荷瘤组织Bcl-2和Cle-Caspase-3蛋白表达水平的变化以及线粒体和细胞质中细胞色素C蛋白含量的变化。结果与模型组相比较,氯喹处理组小鼠移植肉瘤S_(180)生长速度显著减慢,肿瘤体积和瘤质量明显减小(P0.05);透射电子显微镜观察发现,与模型组比较氯喹处理组肿瘤细胞形态出现明显凋亡损伤改变,凋亡小体形成;各剂量组氯喹均可诱导荷瘤鼠S_(180)细胞凋亡,使抗凋亡因子Bcl-2表达下调,凋亡因子Cle-Caspase-3表达上调(P0.05);并且氯喹能够降低线粒体内细胞色素C蛋白的表达,提高细胞质细胞色素C蛋白的表达(P0.05),促使线粒体内细胞色素C向细胞质内释放。结论氯喹能够抑制小鼠S_(180)肉瘤的生长,可能是通过线粒体凋亡途径诱导细胞凋亡,从而发挥抑瘤的作用。     

刺五加多糖对Lewis 荷瘤小鼠抗肿瘤免疫调节作用及机制的研究

目的:探讨刺五加多糖(ASPS)对Lewis 荷瘤小鼠TLR4 信号转导通路的调控机制。方法:用C57BL/6 建立 Lewis 实体型荷瘤小鼠模型,随机分为生理盐水组(NS 组)、阿霉素组(ADM 组)和ASPS 低、中、高剂量组,灌胃给药25 d 后,称 重计算瘤重、抑瘤率、胸腺指数和脾脏指数。收集荷瘤小鼠眼球血,ELISA 检测外周血中细胞因子TNF-α、IL-1β、IL-6 和IL- 12p70 的分泌情况。采用实时荧光定量PCR(Q-PCR)和Western blot 法分别检测荷瘤小鼠脾细胞中TLR4 相关节点TLR4、 MyD88、TRAM、TRAF6、NF-κB p65、AP-1 基因和蛋白的表达水平。结果:与NS 组相比,ASPS 中剂量组荷瘤小鼠的瘤重明显降 低,抑瘤率和免疫器官指数显著升高(P<0.05)。与NS 组相比,ASPS 显著促进荷瘤小鼠外周血TNF-α、IL-1β和IL-6 的分泌 (P<0.05),对IL-12p70 的产生无显著影响(P>0.05)。ASPS 显著上调TLR4 信号通路中TLR4、MyD88、TRAF6、NF鄄资B p65、 AP-1 基因和蛋白在荷瘤小鼠脾脏中的表达(P<0.05),而对TRAM 无显著作用(P>0.05)。结论:TLR4 信号通路可能是ASPS 在Lewis 荷瘤小鼠体内发挥免疫调节及抑制肿瘤作用的通路之一。     

替莫唑胺(TMZ)联合2-甲氧基雌二醇(2-ME)对小鼠胶质瘤化疗作用的体内实验研究

目的:探讨替莫唑胺(temozolomide,TMZ)联合2-甲氧基雌二醇(2-methoxyestradiol,2-ME)对SHG-44胶质瘤的抑制作用。方法:培养SHG-44细胞,建立裸鼠荷瘤模型,荷瘤后将裸鼠随机分空白对照组、TMZ组、2-ME组、TMZ+2-ME组,每组各7只,荷瘤后8 d分别给予TMZ、2-ME、TMZ+2-ME,观察肿瘤的生长情况,给药结束5周后取瘤计算抑瘤率,并制备瘤组织细胞悬液行流式细胞仪检测细胞凋亡率,Western Blot法检测瘤组织HIF-1α、P53蛋白表达。结果:(1)TMZ组、2-ME组及2-ME+TMZ组肿瘤生长均低于空白对照组(P(0.05);2-ME+TMZ组肿瘤生长低于TMZ组。(2)2-ME+TMZ组、TMZ组及2-ME组的瘤细胞凋亡率均高于空白对照组(P(0.05),2-ME+TMZ组与TMZ组的凋亡率相比无统计学差异(P(0.05)。(3)2-ME+TMZ组HIF-1α蛋白表达明显低于TMZ组;2-ME+TMZ组P53蛋白表达明显高于TMZ组。结论:应用替莫唑胺(TMZ)联合2-甲氧基雌二醇(2-ME)对裸鼠皮下移植瘤的治疗作用优于单用TMZ或2-ME。     

Modes of reinforcement and gender and culture of experimenter and subject: effects of "alien" and external reinforcement for Japanese and American men and women

In this study, we examined external and "alien" reinforcement (ER and AR. respectively) as a factor in social learning, and studied the combined effects of culture and reinforcement mode. A female (Experiment 1) and a male (Experiment 2) experimenters conducted experimental sessions. Both men and women, who grew up in the same culture as the experimenter, participated and performed the experimental task. A three-way interaction effect of experimenter gender, culture, and reinforcement mode was found on task performance. And the effect was more pronounced for a Japanese experimenter. A female and a male experimenters conducted Experiments 3 and 4, respectively; however participants this time were men and women who grew up in different cultures than the experimenter. Results indicated that the pattern of the subject gender and reinforcement mode interaction effect, when the experimenter was Japanese with American subjects, was exactly opposite to that when the experimenter was American. These experiments showed that AR was as effective for social learning as ER, and that the cultural backgrounds of experimenter and subject influenced AR and ER effectiveness.     

Rates of oxygen consumption and of anaerobic glycolysis in renal cortex and medulla of adult and new-born rats and guinea-pigs

1. Rates of oxygen uptake and of anaerobic glycolysis were estimated in slices from the renal cortex and medulla (a) of adult rats and guinea-pigs, (b) of new-born (1-, 5- and 21-day-old) rats and of guinea-pigs of 1, 12, 21, 24 and 120 hr age.2. In the adult rat, Q(O2) values for the cortex were 12.55 +/- 0.20 (22) and for the medulla: 8.56 +/- 0.17 (22) mul./hr.mg dry weight, while in the new-born rat (24 hr old) they were 10.99 +/- 0.46 (12) and 9.33 +/- 0.18 (9) mul./hr.mg dry weight respectively.3. Values for Q(CO2) (N2) (anaerobic glycolysis) in the 14 hr old new-born rat were in the renal cortex 9.65 +/- 0.35 (5) and in the medulla 7.39 +/- 0.43 (5) mul./hr.mg dry weight; while in the adult they were 2.25 +/- 0.08 (16) and 5.76 +/- 0.14 (16) mul./hr.mg dry weight, respectively.4. In the adult guinea-pig values for Q(CO2) (N2) were of the same order as in the adult rat, though the rate of O(2) uptake was for the cortex 8.12 +/- 0.22 (12) and for the medulla 5.02 +/- 0.23 (11) mul./hr.mg dry weight.5. Though the Q(O2) values in the renal cortex and medulla were smaller in the 1 hr old new-born guinea-pig, they were already increasing in the 12 hr old neonate.6. The results are discussed in the light of enzyme changes occurring during the process of maturation of the nephron as indicated by histochemical observations.     

Timing and intensity of early fevers and the development of allergies and asthma

BACKGROUND: Early childhood fevers appear to protect against later allergies and asthma. What is not known is the time in which fevers exert this effect and whether the degree of temperature increase is important. OBJECTIVE: We sought to examine the relationship between the time and degree of early fevers and later allergies and asthma. METHODS: Eight hundred thirty-five children from southeast Michigan were enrolled at birth. Clinic records from their first 2 years were abstracted for episodes of fever. At age 6 to 7 years, children underwent allergy testing. We examined fevers occurring within 6-month intervals in the first 2 years of life and outcomes at age 6 to 7 years. The primary outcome measures were allergic sensitization, asthma, asthma with allergic sensitization, and asthma without allergic sensitization. RESULTS: In the unadjusted analysis each episode of fever between 7 and 12 months of age was associated with a lower odds of allergic sensitization (odds ratio [OR], 0.71; 95% CI, 0.54-0.93) and asthma with allergic sensitization (OR, 0.43; 95% CI, 0.21-0.90) at age 6 to 7 years. Likewise, every 1 degrees C increase in the maximum temperature between 7 and 12 months was associated with a lower odds of allergic sensitization (OR, 0.77; 95% CI, 0.61-0.96) and asthma with allergic sensitization (OR, 0.62; 95% CI, 0.40-0.94). After adjusting for potential confounders, each episode of fever between 7 and 12 months was associated with a lower likelihood of asthma with allergic sensitization (adjusted OR, 0.33; 95% CI, 0.11-0.94) at age 6 to 7 years. CONCLUSIONS: Both the timing and intensity of childhood fevers appear to be important factors in the development of allergies and asthma.     

Association between functional alterations of senescence and senility and disorders of gait and balance

OBJECTIVES:

Declines in cognition and mobility are frequently observed in the elderly, and it has been suggested that the appearance of gait disorders in older individuals may constitute a marker of cognitive decline that precedes significant findings in functional performance screening tests. This study sought to evaluate the relationship between functional capacities and gait and balance in an elderly community monitored by the Preventive and Integrated Care Unit of the Hospital Adventista Silvestre in Rio de Janeiro, RJ, Brazil.

METHODS:

Elderly individuals (193 females and 90 males) were submitted to a broad geriatric evaluation, which included the following tests: 1) a performance-oriented mobility assessment (POMA) to evaluate gait; 2) a mini-mental state examination (MMSE); 3) the use of Katz and Lawton scales to assess functional capacity; 4) the application of the geriatric depression scale (GDS); and 5) a mini-nutritional assessment (MNA) scale.

RESULTS:

Reductions in MMSE, Katz and Lawton scores were associated with reductions in POMA scores, and we also observed that significant reductions in POMA scores were present in persons for whom the MMSE and Katz scores did not clearly indicate cognitive dysfunction. We also demonstrated that a decline in the scores obtained with the GDS and MNA scales was associated with a decline in the POMA scores.

CONCLUSIONS:

Considering that significant alterations in the POMA scores were observed prior to the identification of significant alterations in cognitive capacity using either the MMSE or the Katz systems, a prospective study seems warranted to assess the predictive capacity of POMA scores regarding the associated decline in functional capacity.     

男性乳腺发育症临床病理分析及雌,孕激素受体检测

对１１３例男性乳腺发育症进行临床病理分析。同时检测其中３０例乳腺组织中雌激素受体和孕激素受体分布情况，结果发现两者阳性率分别为８０．０％和８３．３３％。结合文献讨论了男性乳腺发育症的发生与高血清激素浓度及乳腺组织高受体水平的关系。     

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分子成像能以非侵入性的方式重现活体细胞的生理功能和生物学过程,提高疾病的早期和特异性诊断水平。纳米颗粒/材料具有物理性质可控性高、易于表面修饰、血液循环时间长和可功能化等优点,在疾病诊断与治疗中显示出巨大潜力。但如何阐明纳米材料多功能间的内在联系、解决其代谢及安全性等关键机制难题、实现纳米颗粒/材料多功能性到临床多功能性的转化,成为目前研究的短板。本文就纳米颗粒/材料在分子成像及诊疗一体化中的应用现状、最新研究进展、面临的挑战和未来前景进行述评。