Expression and characterization of Gag protein of HIV-1（CN） in Pichia pastoris

To express the core protein of HIV-1 of Chinese prevalent strain （HIV-1 （CN）） in Pichia pastoris, the fulllength gag gene was inserted into the secretory expression vector pHILS1. Linearized recombinant plasmid pHILGAG by Sail was electrotransformed into the yeast strain GS115, and the yeast transformants were identified by PCR. To induce the interest protein to be expressed, the PCR positive transformants were inoculated in the medium of BMGY and BMMY, mRNA of the strain was detected by RT-PCR, and the expressed protein was analyzed by SDS-PAGE, Western blotting and thin layer scanning. mRNA （1.3 kb） was amplified by RT-PCR. SDS-PAGE and Western blotting analysis showed that the molecular mass of the expressed protein was 55 kD, which was similar to the expected value, and the expressed protein could react with McAb to HIV-1 p24. Thin layer scanning analysis demonstrated that the whole amount of the expressed protein was approximately 13 % of the soluble protein in the supernatant. The recombinant yeast had good genetic stability. The optimal expression conditions of the engineering yeast were as follows： BMMY medium, 80-90% of dissolved oxygen, 1% methanol, and 3-day-cultivation course. Gag proteins were expressed under the optimal expression condition and purified via gel filtration chromatography. The purity of the interest protein was up to 85 %. After the purified proteins were inoculated into BALB/c mice, the anti-HIV-1 antibodies in the immunized mice could be detected by Western blotting.     

Bioactivity-Guided Isolation of Anticancer Agents from Bauhinia Kockiana Korth

Background

Flowers of Bauhinia kockiana were investigated for their anticancer properties.

Methods

Gallic acid (1), and methyl gallate (2), were isolated via bioassay-directed isolation, and they exhibited anticancer properties towards several cancer cell lines, examined using MTT cell viability assay. Pyrogallol (3) was examined against the same cancer cell lines to deduce the bioactive functional group of the phenolic compounds.

Results

The results showed that the phenolic compounds could exhibit moderate to weak cytotoxicity towards certain cell lines (GI₅₀ 30 – 86 µM), but were inactive towards DU145 prostate cancer cell (GI₅₀ > 100 µM).

Conclusion

It was observed that pyrogallol moiety was one of the essential functional structures of the phenolic compounds in exhibiting anticancer activity. Also, the carboxyl group of compound 1 was also important in anticancer activity. Examination of the PC-3 cells treated with compound 1 using fluorescence microscopy showed that PC-3 cells were killed by apoptosis.     

Linkage of scapuloperoneal spinal muscular atrophy to chromosome 12q24.1-q24.31

Scapuloperoneal (SP) syndromes are heterogeneous neuromuscular disorders which are characterized by weakness in the distribution of shoulder girdle and peroneal muscles. SP syndromes can resemble facioscapulohumeral muscular dystrophy (FSH) due to scapular weakness or Charcot-Marie-Tooth disease (CMT) due to atrophy of peroneal muscles. Both neurogenic and myopathic SP syndromes have been described. Locus for the myopathic form of SP syndrome (scapuloperoneal muscular dystrophy, SPMD) has recently been assigned to chromosome 12q. We previously described a large New England kindred exhibiting an autosomal dominant neurogenic SP syndrome (scapuloperoneal spinal muscular atrophy, SPSMA). Disease expression was more severe and progressive in successive generations, which suggested genetic anticipation. We performed genetic linkage analysis of this family with microsatellite markers and excluded the loci for FSH, CMT, SPMD and SMA (spinal muscular atrophy) in our family. Linkage in our SPSMA family (lod score > 3) was established to seven microsatellite markers that map to chromosome 12q24.1-q24.31. The highest lod score with two-point linkage analysis was 6.67 (theta = 0.00) with marker D12S353. Multipoint analysis gave maximum lod scores of 7.38 between D12S354 and D12S79, and also 7.38 between D12S369 and NOS1 (neuronal nitric oxide synthase). The gene for SPSMA lies within the 19 cM interval between D12S338 and D12S366. This report establishes a locus for the neurogenic form of SP syndrome approximately 20 cM telomeric to the one described for the myopathic form of SP syndrome.      

盐酸西布曲明的合成

目的：改进肥胖症治疗药盐酸西布曲明（ｓｉｂｕｔｒａｍｉｎｅ ｈｙｄｒｏｃｈｌｏｒｉｄｅ）的合成方法。方法：以４－氨苄腈为起始原料,经环烃化、Ｇｒｉｇｎａｒｄ反应、还原、Ｅｓｃｈｗｅｉｌｅｒ－ｃｌａｒｋｅ甲基化和成盐等反应合成了盐酸西布曲明。结果：合成产物经熔点、元素分析、红外光谱和核磁共振谱等确证,总收率３６．３％。结论：此合成工艺简便,易于工业化生产。     

4-O-卤代酰基-4′-去甲基表鬼臼毒素类似物的合成与抗肿瘤活性

为考察4′-去甲表鬼臼毒素C₄位上联结含卤素原子的酯化侧链时对化合物抗肿瘤活性的影响,设计并采用选择性酯化方法合成了9个新的4′-去甲基表鬼臼毒素酯化产物。其中标题化合物在L₁₂₁₀白血病肿瘤细胞与KB细胞的体外生长抑制试验中普遍表现出显著的抑制活性,大部分化合物活性超过依托泊甙。而普通脂酸酯的活性较弱。     

关附甲素的结构简化物及抗心律失常活性

为寻找活性高、毒副作用小的抗心律失常新药,以关附甲素为先导物进行结构简化,共设计合成了1-苯基-3-烷胺基-1,2-丙二醇类化合物14个和吲哚嗪类化合物9个。对抗氯仿诱发小鼠心律失常试验表明,Ⅰ₁,Ⅰ₂,Ⅰ₃,Ⅰ₇,Ⅰ₈,Ⅰ₁₄和Ⅱ27个化合物有较显著的抗心律失常活性,其中Ⅰ₂,Ⅰ₃,Ⅰ₇和Ⅰ₈抗心律失常活性优于关附甲素。     

新生物碱——18-羟基喜树碱

自喜树(Camptotheca acuminata Decne)果中分到一个新生物碱,经光谱分析(UV,IR,¹HNMR,MS,CD)推定为18-羟基喜树碱(Ⅰ),同时分得的还有吕宋果内酯,水杨酸和壬二酸。18-羟基喜树碱对P388白血病细胞有细胞毒活性。     

高位小切口手术治疗精索静脉曲张

目的探讨精索静脉曲张患者高位小切口手术治疗的效果。方法回顾性分析了2005～2006年收治的49例精索静脉曲张患者的病情、治疗及临床效果的有关资料。结果手术治疗49例,仅2例症状体征改变不明显。结论精索静脉曲张患者采用高位小切口手术治疗效果明显,且符合微创要求。     

藻酸双酯钠与蝮蛇抗栓酶-3号对冠心病血液流变学作用的对比观察

对比观察藻酸双酯钠与腹蛇抗栓酶-3号对90例冠心病患者血液流变学作用的影响。结果示:藻酸双酯钠与蝮蛇抗栓酶-3号均能明显改善冠心痛患者血液流变学状态,降低血液粘度。蝮蛇抗栓酶-3号比藻酸双酯钠效果更佳。