Negative feedback inhibition of HIV-1 by TAT-inducible expression of siRNA.

Here we demonstrate that an inducible anti-HIV short hairpin RNA (shRNA) expressed from a Pol II promoter inhibits HIV-1 gene expression in mammalian cells. Our strategy is based on a promoter system in which the HIV-1 LTR is fused to the Drosophila hsp70 minimal heat shock promoter. This system is inducible by HIV-1 TAT, which functions in a negative feedback loop to activate transcription of an shRNA directed against HIV-1 rev. Upon induction the shRNA is processed to an siRNA that guides inhibition of HIV replication in cultured T-lymphocytes and hematopoietic stem cell-derived monocytes. The fusion promoter system may be safer than drug-inducible systems for shRNA-mediated gene therapy against HIV as the shRNAs are only expressed following HIV infection.

[Observation on therapeutic effect of round-sharp needle of new nine-needle and elongated needle for piriformis syndrome with triple puncture method].

OBJECTIVE: To compare the efficacy differences between round-sharp needle of new nine-needle and elongated needle for piriformis syndrome, and explore its action mechanism. METHODS: Eighty cases were randomly divided into a round-sharp needle of new nine-needle group (round-sharp needle group) and an elongated needle treatment group (elongated needle group), 40 cases in each group. The round-sharp needle group was treated with round-sharp needle (0.60 mm x 125 mm) at three points in piriformis with triple puncture method, while the elongated needle group was treated with elongated needle of ordinary specifications (0.32 mm x 125 mm) at three points in piriformis with triple puncture method. Besides, the two groups were also treated with routine acupuncture at Weizhong (BL 40) and Yanglingquan (GB 34), 3 times every week, 2 weeks as one course of treatment. After one course of treatment, the clinical effect was evaluated and the pain threshold values were measured before and after treatment in the two groups. RESULTS: The total effective rate in the round-sharp needle group was 92.5% (37/40), which was superior to 77.5% (31/40) in the elongated needle group (P < 0.05). Compared before treatment, the pain threshold values after treatment in two groups were improved significantly (both P < 0.01). The increment of pain threshold value in the round-sharp needle group was higher than that in the elongated needle group (P < 0.01). CONCLUSION: Round-sharp needle of new nine-needle is effective in treatment of piriformis syndrome and is better than ordinary elongated needle, which is related to that it can effectively increase pain threshold value of the local tissue.

Novel Pol II fusion promoter directs human immunodeficiency virus type 1-inducible coexpression of a short hairpin RNA and protein.

We demonstrate a novel approach for coexpression of a short hairpin RNA (shRNA) with an open reading frame which exploits transcriptional read-through of a minimal polyadenylation signal from a Pol II promoter. We first observed efficient inducible expression of enhanced green fluorescent protein along with an anti-rev shRNA. We took advantage of this observation to test coexpression of the transdominant negative mutant (humanized) of human immunodeficiency type 1 (HIV-1) Rev (huRevM10) along with an anti-rev shRNA via an HIV-1-inducible fusion promoter. The coexpression of the shRNA and transdominant protein resulted in potent, long-term inhibition of HIV-1 gene expression and suppression of shRNA-resistant mutants. This dual expression system has broad-based potential for other shRNA applications, such as cases where simultaneous knockdown of mutant and wild-type transcripts must be accompanied by replacement of the wild-type protein.