MS-222不同处理方式对史氏鲟和中华鲟麻醉效果的影响

MS-222的药液浸浴和药液鳃部喷洒可作为史氏鲟(Acipenser schrenckii)和中华鲟(Acipenser sinensis)的麻醉方式。在水温16～18℃的条件下,史氏鲟的浸浴有效浓度为90～110 mg/L,浸浴安全时间不超过30 min;中华鲟的浸浴有效浓度为80～100 mg/L,浸浴安全时间不超过15 min。史氏鲟药液鳃部喷洒有效浓度为1 500 mg/L,最低有效操作时间为30 s,持续操作时间不超过60 s;药液鳃部喷洒有效浓度为4 000 mg/L,有效操作时间不大于5 s。20～40 mg/L MS-222麻醉液浸浴方式可以作为史氏鲟4 h内的短途运输。史氏鲟在90～150 mg/L的MS-222的药液中超过240 min,试验鱼无法复苏,即生物学死亡。MS-222对中华鲟一般麻醉浓度为80～100 mg/L,其在80 mg/L的MS-222中连续浸浴15 min是安全的。     

稀有鮈鲫麻醉方法研究

在室养条件下观察了MS-222、苯佐卡因和乌拉坦对稀有鮈鲫（Gobiocypris rarus）的麻醉效果。结果表明,乌拉坦不宜作为稀有鮈鲫的麻醉剂,而一定浓度的MS-222或苯佐卡因可导致稀有鮈鲫的快速麻醉;不同麻醉剂浓度、不同的作用时间下,稀有鮈鲫在行为上呈现不同的反应,据此可分为轻度镇静、深度镇静、轻度麻醉、中度麻醉、深度麻醉和髓质麻醉阶段;根据反应时间、恢复时间、维持时间和存活率等,建议使用60mg/L的苯佐卡因或100～110mg/L的MS-222作为深度麻醉剂量。     

MS-222在草鱼、花鲈、鲫鱼组织中的富集与消除

在水温(22±2)℃条件下,将平均体质量约为100 g的健康草鱼、花鲈、鲫鱼在50 mg/L的间氨基苯甲酸乙酯甲磺酸盐(MS-222)药液中浸泡,分别于药浴0.5、1、2、3、7 h和10 h时取样测定肌肉、肝(胰)脏、血液中MS-222的残留量,10 h后将试验鱼置于清洁水中,分别于0.5、1、2、4、6、8、16、32、96、120、192、264、336、432、528、624、720 h时再测定肌肉、肝(胰)脏、血液中MS-222的残留量。结果显示,MS-222在草鱼、花鲈、鲫鱼的血液、肌肉、肝(胰)脏中残留分别在3、3、7 h达到峰值,随后略下降;MS-222在草鱼血液、肌肉、肝胰脏组织中的消除半衰期分别为5.4、6.7、7.9 h;在花鲈血液、肌肉、肝脏组织中的消除半衰期分别为3.9、5.1、5.7 h;在鲫鱼血液、肌肉、肝胰脏组织中的消除半衰期分别为 4.8 、6.3、8.4 h。MS-222在3种鱼类肝(胰)脏、肌肉、血液中的消除速率均为 V肝(胰)脏相似文献   

麻醉剂MS222对尼罗罗非鱼幼鱼耗氧率和排氨率的影响

为探讨MS222在罗非鱼麻醉运输中应用的可能性,在密封式塑料袋中进行了罗非鱼幼鱼的模拟运输。分别用0 mg/L、10 mg/L、20 mg/L和30 mg/L MS222对广特超品系尼罗罗非鱼(Oreochromis niloticus)幼鱼进行处理,在处理0 h、1 h、3 h和7 h后测定各袋水中溶氧量和氨含量,以计算各时段的耗氧率、排氨率及氧氮比。试验结果表明,在7 h的模拟运输过程中,10 mg/L、20 mg/L和30 mg/L MS222可以分别降低罗非鱼的平均耗氧量10.0%、31.4%和40.5%,降低排氮率12.4%、37.8%和42.8%。因此,30 mg/L的MS222更适宜于广特超品系罗非鱼幼鱼运输。     

4种化学投入品对凡纳滨对虾的急性毒性

为了评价硫醚沙星、异噻唑啉酮、鱼安定和杀虫双等4种化学投入品对凡纳滨对虾幼虾毒性风险,在水温24~26℃下,使用静水式生物毒性试验法,测定了以上4种化学投入品对凡纳滨对虾幼虾的急性毒性,确定了四种化学投入品对凡纳滨对虾的半致死浓度和安全浓度。结果表明,硫醚沙星、异噻唑啉酮、鱼安定、杀虫双对凡纳滨对虾幼虾的48 h LC50为0.112、27.838、9.902和56.669 mg/L,对应的安全浓度为0.0003、1.786、1.741和9.210 mg/L。其中,硫醚沙星对凡纳滨对虾毒性较强,具有较大的安全风险。     

Pulsatilla saponin A affects proliferation and radiosensitivity of breast cancer cells by regulating miR-24-3p/RNF2 expression

AIMTo investigate the effect of Pulsatilla saponin A on proliferation and radiosensitivity of breast cancer cells and its mechanism. METHODSHuman breast cancer MCF-7 cells were treated with Pulsatilla saponin A at concentrations of 0, 5, 10, 15 and 20 mg/L and transfected with microRNA-24-3p (miR-24-3p) over-expression vector or inhibitory expression vector. The proliferation and radiosensitivity of the MCF-7 cells were measured by MTT assay and colony formation assay. The miR-24-3p expression and ring finger protein 2 （RNF2） mRNA level were detected by RT-qPCR. The protein expression of RNF2 was determined by Western blot. The luciferase reporter assay was used to detect the targeting relationship between miR-24-3p and RNF2. RESULTSCompared with control group (0 mg/L), the proliferation inhibitory rate of the MCF-7 cells was significantly increased in 5, 10, 15 and 20 mg/L Pulsatilla saponin A groups (P<0.05). The survival score of the MCF-7 cells treated with Pulsatilla saponin A was significantly decreased after irradiation, and the expression of RNF2 was significantly decreased (P<0.05). miR-24-3p targeted RNF2 and negatively regulated its expression. When the MCF-7 cells were simultaneously treated with Pulsatilla saponin A and miR-24-3p, the cell survival curve significantly shifted down. Inhibition of miR-24-3p expression reversed the proliferation-inhibiting and radiation-sensitizing effects of Pulsatilla saponin A on the MCF-7 cells. CONCLUSION Pulsatilla saponin A may affect the proliferation and radiosensitivity of breast cancer cells through miR-24-3p/RNF2 signaling pathway.     

Effect of Herba Taxilli extracts combined with miR-375 on viability and apoptosis of osteoarthritic chondrocytes

AIM To study the effects of extracts of Herba Taxilli (Sangjisheng, SJS) on the viability and apoptosis of osteoarthritic chondrocytes and the underlying mechanism. METHODS Human primary osteoarticular chondrocytes (RPOC) were divided into control group, interleukin-1β (IL-1β) group, IL-1β+low-dose extracts of SJS (SJS-L) group, IL-1β+medium-dose extracts of SJS (SJS-M) group, IL-1β+high-dose extracts of SJS (SJS-H) group, IL-1β+anti-miR-NC group, IL-1β+anti-miR-375 group, IL-1β+SJS-H+miR-NC group, IL-1β+SJS-H+miR-375 group. The cell viability was measured by MTT assay, apoptosis was analyzed by flow cytometry, miR-375 expression was detected by qPCR, and the protein levels of cyclin D1, P21, Bcl-2, Bax and caspase-3 were determined by Western blot. RESULTS Compared with control group, the viability of RPOC at 24 h, 48 h and 72 h and the protein expression levels of cyclin D1 and Bcl-2 were significantly decreased (P<0.05), the protein levels of P21, Bax and caspase-3, the apoptotic rate and the expression level of miR-375 were remarkably increased in IL-1β group(P<0.05). Compared with IL-1β group, the cell viability at 24 h, 48 h and 72 h and the protein expression of cyclin D1 in the RPOC were greatly increased (P<0.05), while the expression of P21 was significantly decreased in IL-1β+SJS-M group and IL-1β+SJS-H group(P<0.05).The apoptotic rate, Bax, caspase-3 protein and miR-375 expression were obviously decreased (P<0.05), and Bcl-2 protein level was significantly increased in IL-1β+SJS-H group compared with IL-1β group(P<0.05). Compared with IL-1β+anti-miR-NC group, the expression of miR-375, the protein levels of P21, Bax, caspase-3 and the apoptotic rate in the RPOC of IL-1β+anti-miR-375 group were markedly decreased (P<0.05), while the cell viability at 24 h, 48 h and 72 h and the protein levels of cyclin D1 and Bcl-2 were significantly increased (P<0.05). Over-expression of miR-375 reversed the effects of extracts of SJS on the viability and apoptosis of RPOC with IL-1β stimulation. CONCLUSION The extracts of Herba Taxilli promotes the viability and inhibits apoptosis of RPOC treated with IL-1β, which is related to the regulation of miR-375 expression.     

Effect of miR-206 over-expression on proliferation and migration of pap?illary thyroid carcinoma K1 cells

AIMTo determine the effect of microRNA-206 (miR-206) on proliferation and migration of human papillary thyroid carcinoma K1 cells and to explore its possible mechanism. METHODSThe expression of miR-206 in the K1 cells was detected by RT-qPCR. The cell viability was detected by CCK-8 assay. The number of viable K1 cells was counted by the method of Trypan blue exclusion. The migration ability of K1 cells was detected by Transwell chamber migration assay. Bioinformatics software was used to predict the target gene of miR-206. The targeting relationship between miR-206 and c-Met was verified by dual-luciferase reporter assay. The protein levels of c-Met, p-Met, AKT, p-AKT, mTOR and p-mTOR were determined by Western blot. RESULTSAfter the K1 cells were transfected with miR-206 mimic transiently, the relative expression of miR-206 in treatment group was significantly higher than that in blank group and negative control group (P<0.01). The results of CCK-8 assay and Trypan blue exclusion assay showed that the proliferation ability of K1 cells in treatment group transfected with miR-206 mimic was significantly inhibited compared with other groups (P<0.01). The results of Transwell assay showed that the number of migratory K1 cells in treatment group was lower than that in blank group and negative control group (P<0.01). Moreover, our results demonstrated that miR-206 directly targeted c-Met and repressed the activation of downstream AKT/mTOR signaling pathway. CONCLUSION miR-206 over-expression inhibits the proliferation and migration abilities of papillary thyroid carcinoma K1 cells, and its mechanism may be related to the inhibition of c-Met/AKT/mTOR signaling pathway.     

Effect of miR-22 secreted by macrophage exosomes on cardiomyocyte autophagy under uremic toxin stimulation

AIM To investigate the effect of microRNA-22 (miR-22) secreted by macrophage exosomes on the autophagy of H9c2 cardiomyocytes under uremic toxin stimulation. METHODS The macrophage-derived exosomes stimulated by indoxyl sulfate (IS) were collected and co-cultured with H9c2 cells. The levels of miR-22 in the macrophages, macrophage-derived exosomes and H9c2 cells were detected by RT-qPCR. The viability of H9c2 cells was measured by CCK-8 assay. The expression of exosome surface marker protein CD63 and autophagy-related proteins LC3 and P62 was determined by Western blot. RESULTS Under IS stimulation, the expression of exosome surface marker protein CD63 in the macrophages was significantly higher than that in control group (P<0.05), and the levels of miR-22 in the macrophages and macrophage-derived exosomes were significantly increased (P<0.01). With the increase in macrophage exosome concentration, the viability of H9c2 cells was decreased gradually (P<0.05), and the stimulation of macrophage exosomes reduced P62 expression and promoted the conversion of LC3-I to LC3-II in a dose-dependent manner (P<0.05). Macrophage-derived exosomes increased the ratio of LC3-II to LC3-I but decreased P62 protein expression in the H9c2 cells transfected with miR-22 mimic compared with the cells transfected with corresponding negative control miRNAs (P<0.05). However, miR-22 inhibitor yielded contrasting results. CONCLUSION IS-stimulated macrophages increase expression of miR-22 in cardiomyocytes through exosomes, and promote autophagy of the cardiomyocytes.