Purpose To assess and compare the antifungal activity of polyhexamethylene biguanide (PHMB), thimerosal, cetylpyridinium chloride, and chlorhexidine, which are disinfectants used in multipurpose disinfectant solutions (MPDSs) against ocular pathogenic Fusarium solani and Aspergillus flavus isolates in vitro. Methods The in vitro activity of PHMB, thimerosal, cetylpyridinium chloride, and chlorhexidine was assessed against 40 isolates of ocular pathogenic fungi that included 24 F. solani and 16 A. flavus isolates. The strains were tested by broth dilution antifungal susceptibility testing of filamentous fungi approved by the CLSI (Clinical and Laboratory Standards Institute) M38-A document. Results MIC90 (minimum inhibitory concentration for 90% of the organisms) values of PHMB were 4 and 16 μg/mL for F. solani and A. flavus, respectively. MIC90 values of thimerosal were 0.0313 and 0.0625 μg/mL for F. solani and A. flavus, respectively. MIC90 values of cetylpyridinium chloride were 2 and 2 μg/mL for F. solani and A. flavus, respectively. MIC90 values of chlorhexidine were 32 and 32 μg/mL for F. solani and A. flavus, respectively. Conclusions As a disinfectant used in MPDSs, thimerosal showed the highest levels of antimicrobial activity against ocular pathogenic F. solani and A. flavus isolates. The concentrations of PHMB (0.0001%), cetylpyridinium chloride (0.00014%), and chlorhexidine (0.003%) in MPDSs are sublethal levels for ocular pathogenic F. solani and A. flavus isolates. Although multiple ingredients within MPDSs play a role in antimicrobial efficacy, antimicrobial activity may be significantly influenced by the disinfectants used in the solution formulations.

PURPOSE Fungal keratitis is emerging as a major cause of vision loss in a developing country such as China because of higher incidence and the unavailability of effective antifungals. It is urgent to explore broad-spectrum antifungals to effectively suppress ocular fungal pathogens, and to develop new antifungal eye drops to combat this vision-threatening infection. The aim of this study is to investigate the antifungal activity of silver nanoparticles (nano-Ag) in comparison with that of natamycin against ocular pathogenic filamentous fungi in vitro. METHODS Susceptibility tests were performed against 216 strains of fungi isolated from patients with fungal keratitis from the Henan Eye Institute in China by broth dilution antifungal susceptibility test of filamentous fungi approved by the Clinical and Laboratory Standards Institute M38-A document. The isolates included 112 Fusarium isolates (82 Fusarium solani species complex, 20 Fusarium verticillioides species complex, and 10 Fusarium oxysporum species complex), 94 Aspergillus isolates (61 Aspergillus flavus species complex, 11 Aspergillus fumigatus species complex, 12 Aspergillus versicolor species complex, and 10 Aspergillus niger species complex), and 10 Alternaria alternata isolates. The minimum inhibitory concentration (MIC) range and mode, the MIC for 50% of the strains tested (MIC50 value), and the MIC90 value were provided for the isolates with the SPSS statistical package. RESULTS MIC50 value of nano-Ag were 1, 0.5, and 0.5 μg/mL for Fusarium spp., Aspergillus spp., and Al. alternata, respectively. MIC90 values of nano-Ag were 1, 1, and 1 μg/mL for Fusarium spp., Aspergillus spp., and Al. alternata, respectively. MIC50 values of natamycin were 4, 32, and 4 μg/mL for Fusarium spp., Aspergillus spp., and Al. alternata, respectively. MIC90 values of natamycin were 8, 32, and 4 μg/mL for Fusarium spp., Aspergillus spp., and Al. alternata, respectively. CONCLUSIONS Nano-Ag, relative to natamycin, exhibits potent in vitro activity against ocular pathogenic filamentous fungi.

In the present study, the antifungal effects of phenylmercuric nitrate and benzalkonium chloride versus those of natamycin and ketoconazole were assessed against 216 filamentous fungi isolates from cases of fungal keratitis. They included 112 Fusarium isolates, 94 Aspergillus isolates, and 10 Alternaria alternata isolates. The strains were tested by broth dilution antifungal susceptibility testing of filamentous fungi approved by the Clinical and Laboratory Standards Institute M38-A document. The results showed that the MIC(50) values of phenylmercuric nitrate were 0.0156, 0.0156, and 0.0313 μg/mL for Fusarium spp., Aspergillus spp., and A. alternata, respectively. The MIC(90) values of phenylmercuric nitrate were 0.0313, 0.0313, and 0.0313 μg/mL for Fusarium spp., Aspergillus spp., and A. alternata, respectively. The MIC(50) values of benzalkonium chloride were 16, 32, and 8 μg/mL for Fusarium spp., Aspergillus spp., and A. alternata, respectively. The MIC(90) values of benzalkonium chloride were 32, 32, and 16 μg/mL for Fusarium spp., Aspergillus spp., and A. alternata, respectively. The study indicates that phenylmercuric nitrate has considerable antifungal activity and its effect is significantly superior to those of benzalkonium chloride, natamycin, and ketoconazole against ocular pathogenic filamentous fungi in vitro, deserving further investigation for treating fungal keratitis as a main drug.

To determine the antifungal activity of phenylmercuric acetate against ocular pathogenic fungi in vitro and develop new antifungal eye drops to combat keratomycosis.The in vitro activity of phenylmercuric acetate was assessed against 261 isolates of ocular pathogenic fungi that included 136 Fusarium spp. isolates, 98 Aspergillus spp. isolates, 10 Alternaria alternata isolates and 17 other pathogens. The activity of phenylmercuric acetate was compared with the activities of amphotericin B and natamycin. In vitro susceptibility testing was performed by broth microdilution assay, in accordance with the CLSI (formerly NCCLS) M38-A guidelines for filamentous fungi.MIC(90)s of phenylmercuric acetate were 0.0156, 0.0156, 0.0156 and 0.0156 mg/L for Fusarium spp., Aspergillus spp., A. alternata and other pathogens, respectively. MIC(90)s of amphotericin B were 2, 2, 1 and 1 mg/L for Fusarium spp., Aspergillus spp., A. alternata and other pathogens, respectively. MIC(90)s of natamycin were 8, 32, 4 and 4 mg/L for Fusarium spp., Aspergillus spp., A. alternata and other pathogens, respectively.Phenylmercuric acetate has promising antifungal activity, which is significantly superior to the activities of amphotericin B and natamycin against a wide variety of ocular pathogenic fungi based on comparative MIC values. Additional evaluation is required to determine its clinical utility.

OBJECTIVE:To investigate antifungal activity of butenafine in comparison with that of natamycin, amphotericin B and fluconazole against ocular pathogenic filamentous fungi in vitro.METHODS:It was an experimental study. Susceptibility tests were performed against 260 isolates of ocular pathogenic filamentous fungi by broth dilution antifungal susceptibility test of filamentous fungi approved by the Clinical and Laboratory Standards Institute (CLSI) M38-A document. The isolates included Fusarium spp. (136), Aspergillus spp. (98), Alternaria alternata (9), Curvularia lunata (3), and unusual ocular pathogens (14). Final concentration ranged from 0.008 to 16.000 mg/L for butenafine, from 0.031 to 16.000 mg/L for amphotericin B and natamycin, and from 0.5 to 256.0 mg/L for fluconazole. Following incubation at 35 degrees C for 48 h, minimal inhibitory concentration (MIC) was determined according to the CLSI M38-A document. For amphotericin B and natamycin, the MIC was defined as the lowest drug concentration that prevented any discernible growth. For butenafine and fluconazole, the MIC was defined as the lowest concentration in which an approximately 75% reduction compared to the growth of the control was observed. Candida parapsilosis ATCC22019 was used as quality control strains to validated the results. Mean MIC and MIC range, the MIC at which 50% of the isolates tested were inhibited (MIC(50)) and the MIC at which 90% of the isolates tested were inhibited (MIC(90)), were provided for all the isolates tested by using descriptive statistical analysis with the statistical SPSS package (version 13.0).RESULTS:MIC(90) of butenafine, natamycin, amphotericin B and fluconazole were 4, 8, 2 and 512 mg/L for Fusarium spp., respectively; 0.063, 32.000, 2.000 and 256.000 mg/L for Aspergillus spp., respectively; 0.5, 8.0, 2.0 and 128.0 mg/L for Alternaria alternate, respectively; 0.125, 2.000, 0.500 and 4.000 mg/L for Curvularia lunata, respectively; and 1, 4, 1 and 256 mg/L for unusual ocular pathogens, respectively.CONCLUSIONS:Butenafine exhibits potent antifungal activity against a wide variety of ocular pathogenic fungi, especially for Aspergillus spp., Alternaria alternata, Curvularia lunata, and some unusual ocular pathogens and may have a role in future studies of antifungal eye drops and treating fungal keratitis.

The in vitro activity of thimerosal versus those of amphotericin B and natamycin was assessed against 244 ocular fungal isolates. The activity of thimerosal against Fusarium spp., Aspergillus spp., and Alternaria alternata was 256 times, 512 times, and 128 times, respectively, greater than that of natamycin and 64 times, 32 times, and 32 times, respectively, greater than that of amphotericin B. Thimerosal's antifungal activity was significantly superior to those of amphotericin B and natamycin against ocular pathogenic fungi in vitro.

ABSTRACT The in vitro activity of the silver nitrat e was assessed in comparison with that of natamycin against 128 corneal Fusarium isolates and 90 corneal Aspergillus isolates. MIC 90 s of silver nitrat e were 2 μg/ml for Fusarium spp. and 1 μg/ml for Aspergillus spp. MIC 90 s of natamycin were 8 μg/ml for Fusarium spp. and 32 μg/ml for Aspergillus spp. Silver nitrat e exhibited potent antifungal activity against ocular fungi in vitro.

OBJECTIVE:To investigate antifungal activity of silver nitrate compared with fluconazole, ketoconazole and amphotericin B against ocular pathogenic fungi in vitro.METHODS:It was an experimental study. Susceptibility tests were performed against 260 isolates (15 genera and 29 species) of ocular pathogenic fungi by broth dilution antifungal susceptibility testing of filamentous fungi (M38-A) approved by National Committee for Clinical Laboratory Standards (NCCLS). Final concentrations ranged from 0.031 to 16.000 mg/L for silver nitrate, ketoconazole and amphotericin B, from 0.5 - 256.0 mg/L for fluconazole. Minimum inhibitory concentration (MIC) was defined as the lowest drug concentration that showed absence of growth or complete growth inhibition (100%). The end points were determined as 100% growth inhibition for silver nitrate and amphotericin B, and > or = 75% growth inhibition for ketoconazole and fluconazole.RESULTS:The MICs at which 90% of isolates were inhibited (MIC(90)) of silver nitrate, ketoconazole, amphotericin B and fluconazole were 2.000, 512.000, 32.000 and 2.000 mg/L for Fusarium species, respectively; 1.000, 256.000, 2.000 and 2.000 mg/L for Aspergillus species, respectively; 2.000, 128.000, 4.000 and 2.000 mg/L for Alternaria alternate, respectively; 2.000, 4.000, 0.125 and 0.500 mg/L for Curvularia lunata, respectively; and 1.000, 256.000, 1.000 and 1.000 mg/L for unusual ocular pathogens, respectively. Silver nitrate was highly active against Aspergillus species (92.9% susceptible at a MIC of < or = 1.0 mg/L) and Fusarium species (96.3% susceptible at a MIC of < or = 2.0 mg/L). 95.6% of Fusarium species and 90.8% of Aspergillus species exhibited resistance to fluconazole, 44.1% of Fusarium species and 42.9% of Aspergillus species exhibited resistance to amphotericin B, 66.2% of Fusarium species exhibited resistance to ketoconazole. The activity of silver nitrate against the fluconazole-resistant, ketoconazole-resistant and amphotericin B-resistant strains was high.CONCLUSION:Silver nitrate has promising activity against a wide variety of ocular pathogenic fungi in vitro, and may have a role in future studies of antifungal eye drops and treating fungal keratitis.

OBJECTIVEThe influence of an antioxidant, propyl gallate (PG), on the activity of amphotericin B (AMB), terbinafine (TBF), butenafine (BTF) and ketoconazole (KCZ) against ocular pathogenic filamentous fungi in vitro was investigated to determine whether PG could increase the antifungal activity.METHODSSusceptibility tests were performed against 6 isolates of ocular pathogenic filamentous fungi (Fusarium solanae, Fusarium moniliforme, Fusarium poae, Fusarium oxysporum, Aspergillus fumigatus and Aspergillus flavus) and 2 quality control strains (Candida krusei ATCC 6258 and Cadida parapsilosis ATCC 22019) by the NCCLS M38-P broth microdilution method (MIC). PG was added to the incubation media at a final concentration of 400 microg/ml. Antifungal agents were serially two-fold diluted and final dilutions were made in 1640 and PG-1640 culture media to a concentration ranging from 0.0313 to 16 microg/ml for AMB, TBF, BTF and KCZ. One hundred microl of the corresponding diluted inoculum suspension was added to each well of the microdilution tray. The MIC end-point of AMB was determined as 100% growth reduction and the MIC end-point of TBF, BTF, KCZ and PG was determined as 75% growth reduction as compared with the turbidity produced by the control well.RESULTSAt a concentration of 400 microg/ml, PG did not show any antifungal activity under these experimental conditions. The combination of PG (400 microg/ml) with amphotericin B revealed a remarkably increased activity against all of the isolates of ocular pathogenic filamentous fungi and quality control strains. In the combination of PG with terbinafine, a remarkably increased activity was observed against Fusarium solanae, Fusarium poae, Fusarium oxysporum, Aspergillus fumigatu and Aspergillus flavus. The combination of PG with butenafine had remarkably synergistic effect against Fusarium solanae, but did not synergistic or even showed antagonistic effect for other isolates. The combination of PG with ketaconazole was synergistic against Fusarium solanae, but was antagonistic against all other isolates.CONCLUSIONSCombination of PG and amphotericin has remarkably synergistic effect against all tested ocular pathogenic filamentous fungi isolates. Combination of PG and terbinafine has remarkably synergistic effect against some isolates. The PG-amphotericin combination and the PG-terbinafine combination may have a role in future studies of antifungal eye drops.

噻吗洛尔是一个有效的降眼压药物并且已经用于治疗慢性开角型青光眼的治疗近30年了[1].即使到目前噻吗洛尔仍然是最重要的抗青光眼药物之一,然而普通的滴眼液眼部的生物利用度非常低,大部分都被鼻泪管排泄掉了[2].

OBJECTIVE To evaluate the antifungal activities of penetration enhancer laurocapram(Azone) against ocular pathogenic filamentous fungi in vitro.METHODS The minimum inhibitory concentration (MIC) of Azone against 4 strains of ocular pathogenic filamentous fungi (Fusarium moniliforme, Aspergillus flavus, Aspergillus fumigatus, and Aspegillus niger) were determined by broth microdilution method.RESULTS MICs of Azone were 0.003 1 μL·mL -1 against Fusarium monilifore and Aspergillus flavus, 0.006 3 μL·mL -1 against Aspergillus fumigatus, 0.012 5 μL·mL -1 against Aspergillus niger, respectively.CONCLUSION Azone was active against ocular pathogenic filamentous fungi in vitro. It suggested that Azone incorporated into the ophthalmic formulations of antifungal drugs had two functions. One was promoting the cornea absorption of antifungal drugs,and the other was cooperating antifungal activities with antifungal drugs. The two functions of Azone will increase therapeutic action of antifungal agents.

OBJECTIVE:To compare the pharmacokinetics of 0.5% pilocarpine containing sodium hyaluronate with 1% generic pilocarpine solution.METHODS:One hundred albino rabbits were divided into 20 groups, each consisting of 5 animals. Ten groups received 0.5% pilocarpine containing sodium hyaluronate and 10 groups received 1% generic pilocarpine solution as control. The aqueous humor was withdrawn at 5, 10, 20, 30, 40, 60, 90, 120, 150, and 180 min after instillation. The drug was extracted from aqueous humor with dichloromethane and was detected by reversed phase high performance liquid chromatography (HPLC).RESULTS:The average recovery rate of pilocarpine from aqueous humor was 98.2%. The minimum detectable concentration was 0.025 micro g/ml. The peak concentration and half-life of pilocarpine in aqueous humor were 4.46 micro g/ml at 10 min and 31.83 min, respectively, in the experimental group. Whereas, the peak concentration and half-life of pilocarpine in aqueous humor were 2.25 micro g/ml at 20 min and 22.98 min, respectively, in the control group. The peak concentration of pilocarpine in aqueous humor in the experimental group was 1.98 (P < 0.05) times higher than the control group. The area under curve of the drug concentration-time (AUC(0 - 180)) in the experimental group was 1.75 times higher than the control group.CONCLUSION:Pilocarpine (0.5%) containing sodium hyaluronate significantly increased the peak concentration of pilocarpine, shortened the time of reaching peak concentration and prolonged the half-life in aqueous humor. These results indicate that 0.5% pilocarpine with sodium hyaluronate significantly increases ocular bioavailability of pilocarpine.

目的 评价两种自制的增效酮康唑滴眼液及酮康唑混悬滴眼液体外抗眼科丝状真菌活性.方法用微量液基稀释法测定两种增效酮康唑滴眼液及酮康唑混悬滴眼液对4株眼科常见致病丝状真菌(串珠镰刀菌、烟曲霉菌、黄曲霉菌及黑曲霉菌)的最低抑菌浓度(MIC),用二甲基亚砜(DMSO)溶解的酮康唑液做对照.结果增效酮康唑滴眼液Ⅰ、增效酮康唑滴眼液Ⅱ、酮康唑混悬滴眼液及DMSO溶解的酮康唑液对串珠镰刀菌的MIC分别为2、1、16、2μg/mL;对烟曲霉菌的MIC分别为0.5、0.5、2、0.5μg/mL;对黄曲霉菌的MIC分别为16、16、128、16μg/mL;对黑曲霉菌的MIC分别为0.25、0.25、4、0.25μg/mL.上述结果显示3种酮康唑滴眼液及DMSO溶解的酮康唑液对黑曲霉菌的作用最强,对烟曲霉菌、串珠镰刀菌的作用依次递减,对黄曲霉菌的作用较弱.两种增效酮康唑滴眼液的作用强度与DMSO溶解的酮康唑液的相似,三者的抗真菌活性均明显强于酮康唑混悬滴眼液.结论两种增效酮康唑滴眼液对4株眼科常见致病丝状真菌的体外抗真菌活性较酮康唑混悬滴眼液的明显增强,说明增加酮康唑的溶解度可明显增加其抗真菌活性。

目的 探讨建立高眼压模型的方法,观察急性高眼压诱导的眼组织的脂质过氧化反应.方法采用α-糜蛋白酶-水门汀及2%甲基纤维素二种诱导剂于26只家兔前房注入梗阻房角的方法建立高眼压模型,持续高眼压2周后将对照眼及实验眼制备匀浆测定NO、NOS活性及MDA水平,同步测定F-ERG a、b波振幅的变化及视神经纤维层参数分析,参照病理学改变探讨高眼压的毒理作用.结果形态学和功能学检查表明高眼压模型建立成功,NO水平在晶状体及脉络膜组织中普遍增高,尤其显著的是视网膜内NO释放上升(P＜0.05),NOS活性轻度增高.匀浆内MDA在晶状体及视网膜内显著升高(P＜0.01).该结果与青光眼视神经节细胞凋亡密切相关.结论高眼压诱导眼内组织NO水平升高,MDA水平显著升高,证实视网膜内具有两个毒性底物(NO、MDA)将诱导形成毒性更强的凋亡剂--过氧亚硝酸阴离子.提示临床给予抗氧化剂治疗青光眼很有必要。

派立明是一种最新型的局部碳酸酐酶抑制剂.此药可选择性、高亲和力及明显地抑制碳酸酐酶同功酶Ⅱ的活性,有效地降低眼压.本品滴眼后可快速进入眼组织,在虹膜、睫状体、脉络膜、视网膜、晶状体和血液中有较长的半衰期(数天).虽然用派立明滴眼后,可在全血中测出药物浓度,提示该药可全身吸收,但主药和其代谢产物的血浆浓度非常低,在稳定状态下药物与红细胞内碳酸酐酶的结合达不到完全饱和.因此,不会出现全身酸中毒或其他与口服碳酸酐酶抑制剂有关的副作用.对兔眼滴用派立明还可增加视乳头血流量,而对全身酸碱平衡的影响极小.如这一作用在人眼被证实,将对有视神经病变的青光眼患者十分有益.1%派立明每日滴眼2次的降眼压效果最好,且患者的耐受性较多佐胺好,这可提高患者长期用药的依从性.滴眼后最常见的副作用是视物模糊(6%)及口苦、口酸等味觉异常(6%).总之,派立明的降眼压作用强,副作用小,滴眼舒适,患者耐受性好,是一种非常有价值的抗青光眼新药.

ObjectiveTo establish the model of microcirculation obstacle of ocular conjunctiva in rabbit and observe the effect of MMH eye drop on microcirculation obstacle model and appraise the possibility of improvement in microcirculation obstacle by MMH eye drop.MethodThe model of microcirculation obstacle of conjunctiva was made by injecting high molecular dextran(molecular weight 21×104)into ear vein of rabbit.0.9% Sodium chloride was used in model group; middle and Low MMH consistence drops were used in experimental groups.The shape integral flowing shape integral and surroundings of blood capillary integral,etc were automatically analysed by WX-8 microcirculation instrument(Contain picture of automatic management system).ResultIn normal eye of rabbit the intersection number of blood capillary were increased and the speed of blood flow was quick after using MMH eye drop(P<0.05～0.001)In model group microcirculation obstacal was more serious than that in the MMH groups(P<0.05～0.001).ConclusionThe model of microcirculation obstacle of conjunctiva was successfully established in rabbit.The results suggest that MMH eye drop can prevent conjunctiva from mirocirculation abstacle

目的建立鸡眼形觉剥夺性近视(FDM)模型,观察复方珍珠水解液(MPHS)药物防治近视的疗效.方法建立FDM模型,艾维茵仔鸡左眼鼻侧贴上半透明新月形1/2眼睑眼罩,造成1/2眼球形觉剥夺;采用A超,测定双眼眼内结构;KR7100型电脑自动验光仪测量各组双眼屈光度;采用非接触式阿贝比长仪测定各眼球外径;正位照片测量赤道8个半径.通过眼球形态学、屈光学比较FDM模型组与各组间的差异.结果模型组比正常组眼球外径扩大.玻璃体腔扩张为113%;眼内轴长增大至108%;8个赤道半径扩大,差异呈显著性(P＜0.05～0.001).屈光度增加-2.087 D;而MPHS药品则显著抑制了眼球形态学的扩张;抑制负性屈光度的增长.结论(1)FDM模型眼球的各种测量结果均证实模型建立成功,玻璃体腔明显扩张至130%,眼内轴长延长至108%,通过8个赤道半径测量全面扩大.(2)MPHS具有抑制眼球外径、内径及赤道半径扩张的作用.

目的 探讨氧化应力、5种抗白内障药物对晶状体一氧化氮合酶 (NOS)活性及一氧化氮 (NO)水平的影响。方法 建立晶状体温育模型 :培养液分 7组 :( 1)对照组含DMEM 10ml;( 2 )～ ( 7)组含DMEM 10ml外尚有 3 0 %H2 O2 0 2ml ,FeCL3 2mg ,并于 ( 3 )～ ( 7)组中分别加入海珠神、麝珠明目、益视安、晶福及视明露滴眼液各 1 0ml ,晶状体制备匀浆上清 ,测NOS ,NO ,MDA。结果 1.正常人、兔、SD鼠晶状体的NOS活性分别为 ( 6 45± 1 5 7)、( 18 0 3± 8 2 0 )、( 13 61± 2 13 )U/mgpro .,人、兔晶状体内均测不出NO ,而SD鼠NO为 ( 5 2 79± 2 0 79) μmol/mgpro .。 2 .氧自由基诱导温育晶状体NOS活性上升 1 5倍 (P <0 0 0 1) ,但晶状体内测不出NO。 3 .抗白内障药物影响NOS活性及NO水平 ,海珠神、麝珠明目、益视安及晶福组与氧化组相比显示抑制NOS活性 ,分别下降 15 % ,9% ,6%及 5 5 % ,而视明露组诱导NOS活性上升 41% (P <0 0 0 1)。 4.氧化组晶状体MDA上升 2 15倍 ,各抗白内障药物组MDA下降至对照组以下 ,证明 5种抗白内障药物都是抗氧化剂。结论 氧自由基诱导晶状体NOS活性上升 ,NO测不出。NO与白内障是否有关尚待深入研究。

It is kno wn that 8-Br-cAMP is one of selective bi nding site analogues for cAMP RIIα to af fect cell growth through regulation of g ene expression.The p16,p21wafl,p53 a nd Rb are antioncogenes which affect cel l growth through control of cell cycle.T he aim of this study is to investigate t he 8-Br-cAMP effect on the expression of antioncogenes in human HXO-Rb44 cells. Methods Cultured HXO-Rb44 cells in RPMI -1 640 medium were divided into two aliquot s.8-Br-cAMP (2×10-5mol/L) was added i nto one aliquot for 24h as the experime ntal group(EG),the another aliquot witho ut 8-Br-cAMP as the control group(CG).Af ter 24h,the cell suspension was dropped onto the nitrocellulose membrane.The mR NA of p16,p21wafl,wild type(w)p53,mut ant type(m) p53 and Rb were used respec tively with biotin-labeled cDNA probes b y intact cell RNA dot blot.The immunorea ctivity(IR) of P16,P21wafl,PRb,PCN A,cdk2 and cdk4 were detected respecti vely with specific monoclonal antibodies on dot blot.ResultsThe mRNA dot blot s ignals of mp53 and protein dot blot of cdk2-IR,cdk4-IR and PCNA-IR in EG were weaker than those in CG(P＜0.05～0.01). W hile,the mRNA signals of p16,p21wafl,wp53 and Rb in EG were stronger than tho se in CG(P＜0.05～0.01).The intensity of ea ch protein dot blot was consistent with that of their RNA dot blot (except for w P53-IR and mP53-IR not to be done).Conc lusions(1)8-Br-cAMP could up-regul ate expression of antioncogenes includin g p16,p21wafl,wp53,Rb,and protein exp ression of P16,P21wafl and PRb.(2) 8-Br-cAMP could down-regulate mp53 gene expression and protein expression of cd k2,cdk4 and PCNA.The results suggest t hat 8-Br-cAMP could inhibit human HXO-Rb 44 cell growth through interfering rela ted gene expression of cell cycle. Key words: retinoblastoma; antioncogenes; dot; blot; cdks