通过原生质体融合获得青霉素高产菌株

采用两株分别带有不同抗药性标记的产黄青霉原生质体以1:1相混合,用30％的聚乙二醇(MW6000)原生质体融合处理。将混和物涂布在补给有两种药物同时存在的高渗性分离培养基上,培养后获得异核体菌落。洗下异核体分生孢于继续分离在补给有两种药物同时存在的分离培养基上,得到了同时带有两种抗性标记的重组体。纯化后,进行抗性稳定性和摇瓶效价的测定,将一株具有双重抗性、摇瓶发酵效价较高的菌株作出发菌株,经诱变因子处理后,获得了一株43u7c—57菌株,在摇瓶中效价比生产菌株平均提高4.94％,该菌株在40吨发酵罐试生产后,发酵效价平均提高4.4％,产量则平均增加12.15％。     

Accuracy of allometrically predicted pharmacokinetic parameters in humans: role of species selection.

A general equation was derived, which directly describes the mathematical relationship between the allometrically predicted pharmacokinetic (PK) parameters in humans and the body weights of animal species (along with their corresponding measured PK parameters). It was shown, with use of the derived equation, that the predicted values in humans, based on combinations of animal species commonly used in allometry, are heavily dependent on certain species, for example, the dog. In contrast, parameter values from the rat made no contribution to the predicted human values, as long as the rat was not the smallest species used. Monte Carlo simulations were further performed to examine the species or weight dependence. The cost-effective combinations of animal species, in terms of number and species type, were theoretically examined through simulations. Finally, literature data demonstrated the species or weight dependence predicted from the equation and as illustrated through the Monte Carlo simulations. Appreciation of this species or weight dependence should guide researchers in selecting animal species and designing optimal experiments in the application of allometric scaling.     

应用改进的AdEasy系统制备重组腺病毒

目的 改进AdEasy系统，使之更简便、高效、快捷。方法 pAdtrack经酶切线性化后，转化经氯化钙处理的感受态AdEasier-1细菌，获得重组腺病毒质粒。最后将线性化的重组腺病毒质粒转染293细胞包装获得重组腺病毒。荧光显微镜下细胞中重组腺病毒观察绿色荧光蛋白的表达。结果 应用氯化钙法由pAdtrack转化AdEasier-1细菌，可获得极高比例的(20／20)阳性重组腺病毒克隆。结论 应用改进后的AdEasy系统构建腺病毒相比于传统AdEasy系统，方法更简便、成功率更高、实验周期更短、对实验室的条件要求低。     

Soluble polystyrene derivatives of metal bis(3,4-dicarboxybenzoyl)phthalocyaninates: Synthesis,thermal analysis,and sensitivity towards toxic gases

Polystyrene-bound metal [2,9 or 2,10 (or 2,16 or 2,17) bis(3,4-dicarboxybenzoyl)]phthalocyaninates were synthesized by Friedel-Crafts reaction of polystyrene with the corresponding metal phthalocyaninates. Co(II) and Cu(II) [2,9 or 2,10 (or 2,16 or 2,17) bis(3,4-dicarboxybenzoyl)]-phthalocyaninate (PS-CodaPc and PS-CudaPc) contained 0,13 mmol · g^?1 (12,4 wt.-%) and 0,13 mmol · g^?1 (12,8 wt.-%) of CodaPc and CudaPc, respectively. They were soluble in N,N'-dimethylformamide, but only partially soluble in chloroform, tetrahydrofuran (THF), dimethyl sulfoxide, N-methyl-2-pyrrolidone, and pyridine. The THF extracts contained 0,12 mmol · g^?1 (11,4 wt.-%) and 0,18 mmol ? g^?1 (17,2 wt.-%) of PS-CodaPc and PS-CudaPc, respectively. The thermal stability of the polymers was studied using thermogravimetric and differential thermal analysis in nitrogen and synthetic air atmosphere. The contents of MdaPc(M: metal) in THF-extracted polymers calculated from the data of residue in thermogravimetric analysis are 0,12 mmol · g^?1 for PS-CodaPc and 0,19 mmol · g^?1 for PS-CudaPc. In addition, the sensitive properties of the polymers towards toxic gases were also investigated by quartz microbalance transducers. The results show that the quartz microbalance sensors coated with both polymers were sensitive to NO₂ and chlorinated hydrocarbons, i.e. chloroform and perchloroethylene. The sensitivity to NO₂ was 6,53 · 10^?7 m³ · mL^?1 · s^?1 for PS-CodaPc and 1,90 · 10^?6 m³ · mL^?1 · s^?1 for PS-CudaPc, and that to chloroform and perchloroethylene was 2,33 · 10^?8 and 4,60 · 10^?8 m³ · mL^?1 · s^?1, respectively, for PS-CodaPc and 4,79 · 10^?8 and 9,51 · 10^?7 m³ · mL^?1 · s^?1 for PS-CudaPc.     

慢粒白血病abl癌基因表达的研究

选择分别位于ｂｃｒ／ａｂｌ嵌合基因的Ｍｂｃｒｌ第二外显子和ａｂｌ的第二外显子上的两条引物，对１８例慢粒白血病及２例急淋白血病标本进行逆转录ＰＣＲ（ＲＴ－ＰＣＲ）检测。结果：在１８例包括ｐｈ（＋）和ｐｈ（－）不同病期的慢粒白血病患者血中均检出ｂｃｒ／ａｂｌ嵌合基因的表达，其中１４例为Ｋ－２８型表达，１例为Ｌ－６型表达，３例为Ｋ－２８型，Ｌ－６型同时表达；２例急淋白血病标本中１例为Ｋ－２８型表达。研究结果表明，１．ｂｃｒ／ａｂｌ嵌合基因是慢粒白血病的一个重要分子生物学特征；２．ｐｈ（＋）和ｐｈ（－）慢粒白血病的分子生物学基础相同，即均有ｂｃｒ／ａｂｌ嵌合基因；３．至少部分急淋白血病与慢粒白血病有相同的分子生物学基础；４．ｂｃｒ的断裂点位置可能有一定的临床意义，但有待进一步研究；５．同时表达Ｌ－６型和Ｋ－２８型的情况多见于急变区，此现象提示体内可能同时有两类不同嵌合的白血病细胞或白血病急变期ｂｃｒ出现新的重排。     

甾体避孕药不增加血栓栓塞性疾病危险(病例对照研究)

本文采用1:3病例对照研究方法,对甾体避孕药的使用与血栓栓塞性疾病(急性心肌梗塞、脑栓塞、脑血栓形成和深静脉血栓形成)间的关联进行多因素分析。结果提示在我国广泛应用的甾体避孕药(主要为国产Ⅰ号避孕药)并不增加血栓栓塞性疾病的危险性。用药和停药时间的长短与血栓栓塞性疾病的发生无关,并与高血压病之间亦不存在协同作用。     

穿透性角膜移植术后角膜内皮细胞之命运

地２组猴行５。５→５．０ｍｍ穿透性角膜移植术，异种组４只，同种组４只，术前用冷冻创伤＾３Ｈ－ＴｄＲ标记法标记受生角膜内皮细胞，术后１４周摘除眼球，用放射自显影法显示银颗粒的分布。     

Monitoring the protective effects of minocycline treatment with radiolabeled annexin V in an experimental model of focal cerebral ischemia.

Minocycline is an antibiotic now recognized to have antiapoptotic and antiinflammatory properties. Because of these properties, minocycline may be of benefit in reducing neuronal apoptosis from ischemia and subsequent postischemic inflammation if administered soon after a stroke. We now explore the feasibility of using (99m)Tc-annexin V, an in vivo marker of apoptosis, with SPECT to monitor the antiapoptotic effects of minocycline therapy. METHODS: CB6/F1 adult male mice underwent unilateral distal middle cerebral artery occlusion (dMCA) occlusion and were imaged and sacrificed at 1, 3, 7, or 30 d after injury. Animals were given minocycline (or vehicle) 30 min and 12 h after dMCA occlusion and then given 22.5 mg/kg twice daily for up to 7 d. Before imaging, behavioral tests were performed to evaluate the neurologic function. After imaging, brains were collected for histology and assessed for the degree of apoptosis and microglial activation. RESULTS: (99m)Tc-Annexin V uptake in injured hemispheres was significantly decreased 2- to 3-fold by minocycline at all time points. Minocyline reduced infarct size as seen histologically and improved behavioral indices as late as 30 d. Infarct volume as seen histologically correlated with radiolabeled annexin V uptake seen by SPECT. In situ fluorescent microscopy demonstrated that annexin V bound primarily to neurons at 1 and 3 d, with a shift toward microglia by 7 and 30 d. CONCLUSION: We found that minocycline significantly reduces neuronal apoptosis and infarct size and improves neurologic outcome in mice after acute focal cortical ischemia.