Rhizoma Pinelliae Fermentata(RPF)wasoneofthecommonlyusedChinesemateriamedica(CMM). According to the ancient and modern literatures on RPF, the historical evolution, fermentation methods, chemical compositions, efficacy and microbes of RPF were systematically summarized in this paper. Through the analysis on existing problems of fermentation strains, effective components, quality standard and fermentation process, the corresponding solutions were proposed. This work may provide an idea and reference for the further study of RPF.

Objective To establish a recombinant Mycobacterium (M.) smegmatis secreting interliukin-2 (IL-2) for the prevention and treatment of human bladder cancer. Methods The M. tuberculosis HSP70 promotor, ?-antigen signal peptide gene, and human IL-2 cDNA were amplified from plasmid pY6013, pIJK-1, and pHIG53 respectively by PCR. The products were cloned into plasmid pRR3 to construct a mycobacterial shuttle-expressing plasmid pR-a-IL-2 secreting human IL-2. After confirming the construction was correct by enzyme digestion, plasmid pR-?-IL-2 was transduced into M. smegmatis mc 2 155 by electroporation. The stability of the recombinant mycobacteria was evaluated and the activity of IL-2 secreted by the bacteria was assayed. Results Structure of the pR-a-IL-2 was correct and it was effectively transduced into M. smegatis mc 2 155. The recombinant mycobacteria stably expressed IL-2. The IL-2 activity in the medium was 118.5 U/ml. Conclusion The successful establishment of recombinant M. smegmatis can provide the basis for the research of biotherapy and prevention of the recurrence of bladder cancer.

It is generally agreed that the central cholinergic system plays an important role in learning and memory processes. Alzheimers disease (AD), the most common cause of dementia, is a devastating illness characterized by progressive cognitive deterioration. The learning and cognitive deficits observed in AD patients are hypothesized to be partly caused by central cholinergic system dysfunction. There exists a malicious cycle in the pathologic course of AD, that is, the impairment of central cholinergic transmission could increase the amount of A? in the brain, and the increase of A? could also impair the efficacy of the cholinergic transmission.

Alpha-toxin gene was amplified from chromosomal DNA of Clostridium perfringens type C by polymerase chain reaction(PCR). PCR product was inserted into vector pGEM-T directively. The cloned recombinant plasmid pXCPA1 possesses positive nucleotide sequence of alpha-toxin. A 1.2 kb alpha-toxin gene fragment was cleaved with restriction endonucleases NcoI/EcoRI from plasmid pXCPA1, and then inserted into an expression vector pET-28c which cleaved with NcoI/EcoRI by blunt-end ligation. The recombinant expression plasmid pETXA1 was studied in detail by restriction endonucleases analysis and nucleotide sequencing. The results showed that the recombinant expression pETXA1 possessed a positive alpha-toxin gene sequence and reading frame. BL21(DE3) (pETXA1) could produce alpha-toxin and the expressed products were recognized by alpha-toxin monoclonal antibodies, and the expression level of the alpha-toxin proteins were about 16.28% of total cellular protein by SDS-PAGE and thin-layer gel scanning analysis.