局部联合外用1％阿达帕林（达芙文）和单纯外用1％克林霉素治疗痤疮的疗效比较以及局部外用1％阿达帕林（达芙文）进行维持治疗的疗效观察

比较局部联合外用1％阿达帕林(达芙文)和1％克林霉素治疗痤疮的效果，并观察单纯外用1％阿达帕林(达芙文)进行维持治疗的效果。方法：共有300名Ⅱ—Ⅲ痤疮患者随机分成联合用药组和单纯克林霉素组，共治疗12周。联合用药组联合使用1％阿达帕林(达芙文)和1％克林霉素，对照组单纯使用1％克林霉素。联合用药组早晨外用一次1％克林毒素，晚上先用1％阿达帕林，2分钟后再使用1％克林霉素，而对照组早、晚各外用1％克林霉素一次。在维持治疗的疗效观察中，治疗组每晚外用1％阿达帕林一次共12周，对照组不用药。治疗开始和每次随方时，计算皮损总数、非炎症性皮损的数量和炎症性皮损的数量。结果：联合用药组的治疗效果较好。在维持治疗中，与第12周相比较，治疗组在第24周的皮损总数、非炎症性皮损的数量和炎症性皮损的数量分别下降了41．6％，41．7％ and 40．8％，而对照组分别上升了92．1％，97．1％ and 87．7％。结论：与单纯外用1％克林霉素相比，联合使用1％阿达帕林(达芙文)和1％克林霉素治疗痤疮在减少皮损数量等方向效果更好。在常规治疗后，需要用1％阿达帕林(达芙文)进行维持治疗。

Cloning of HBsAg-encoded genes in different vectors and their expression in eukaryotic cells

AIM: To compare the efficiency of different plasmids as DNA vectors by cloning three HBsAg-encoded genes into two eukaryotic expression vectors, pRc/CMV and pSG5UTPL/Flag, and to express HBsAg S, MS, and LS proteins in SP2/0 cells, and to establish monoclone SP2/0 cell strains that are capable of expressing S or S2S proteins stably.METHODS: Segments of S, preS2-S, preS1-preS2-S genes of Hepatitis B virus were amplified by routine PCR and preS1S fragment was amplified by Over-Lap Extension PCR. The amplified segments were cleaved with restricted endonuclease Hind Ⅲ/Not Ⅰ followed by ligation with pRc/CMV, or BamHI/EcoR Ⅰ followed by ligation with pSG5UTPL/Flag. After the plasmid vectors were cleaved with the correspond enzymes, the amplified segments were inserted into pRc/CMV or pSGSUTPL/Flag plasmid vectors with T4DNA ligase. KOZAK sequence was added before the initial ATG code of each fragment using specific primer. The inserted segments in the recombinant plasmids were sequenced after subcloning. BALB/c mice myeloma cells (SP2/0 cell line) were transfected with the recombinant plasmids. The expressions of the different recombinants were compared by Western-blot, using a monoclonal anti-HBs antibody as the primary antibody and peroxidase-labeled multi-linker as the secondary. Stable SP2/0-pRc/CMV-S or SP2/0- pRc/CMV-MS clones were established through clone screening with G418.RESULTS: Fragments with anticipated size were harvested after PCR. After recombination and screening, the sequences of the inserted segments in the recombinants were confirmed to be S, preS2S, preSl-preS2S and preSlS encoding genes,determined by sequencing. The results of Western-blot hybridization were positive for the anticipated proteins.Among them, pRc/CMV-S or pRc/CMV-MS demonstrated the highest expressing their respective antigen.CONCLUSION: Eight recombinant plasmids expressing S,M, L or preSlS proteins are obtained. For hepatitis surface antigen expression in eukaryotic cells, the vector pRc/CMV is superior to pSG5UTPL/Flag, and pRc/CMV-S and pRc/CMVMS are the most efficient in the pRc/CMV clones. SP2/0 cells stably expressing HBsAg are established, and may be used as target cells for evaluating the CTL activity of a DNA vaccine in vitro.

The host type I interferon response to viral and bacterial infections

Type I interferons (IFN) are well studied cytokines with anti-viral and immune-modulating functions. Type I IFNsare produced following viral infections, but until recently, the mechanisms of viral recognition leading to IFN productionwere largely unknown. Toll like receptors (TLRs) have emerged as key transducers of type I IFN during viral infectionsby recognizing various viral components. Furthermore, much progress has been made in defining the signaling path-ways downstream of TLRs for type I IFN production. TLR7 and TLR9 have become apparent as universally importantin inducing type I IFN during infection with most viruses, particularly by plasmacytoid dendritic cells. New intracellularviral pattern recognition receptors leading to type I IFN production have been identified. Many bacteria can also inducethe up-regulation of these cytokines. Interestingly, recent studies have found a detrimental effect on host cells if type IIFN is produced during infection with the intracellular gram-positive bacterial pathogen, Listeria monocytogenes. Thisreview will discuss the recent advances made in defining the signaling pathways leading to type I IFN production.