Promote DC maturation and migration; and increase the secretion or expression of cytokines and co-stimulatory molecules such as CD80, CD86, and MHC-II. Here, we ascertained the ability of PA-MSHA to activate innate immune responses through assessing TLR signaling pathway activation in splenocytes and BMDCs activation following in vitro stimulation with PA-MSHA. In this study, we assessed the ability of PA-MSHA to activate innate immune responses in murine splenocytes and BMDCs, as well as its in vivo adjuvant effects in enhancing cellular and humoral immune responses to HIV-1 Env peptides following co-administration with a DNA vaccine. PA-MSHA enabled activation of the TLR pathway mediated by NF-kB and JNK signaling in splenocytes, and promoted the up-regulation of co-stimulatory molecule CD86 in BMDCs. As well, co-inoculation of the DNA vaccine with low dosages of PA-MSHA enhanced specific immunoreactivity against HIV-1 Env in both cellular and humoral responses, and promoted antibody avidity maturation. However, high doses of adjuvant resulted in an immunosuppressive effect; a two- or three-inoculation regimen yielded low antibody responses and the two-inoculation regimen exhibited only a slight cellular immunity response. To our knowledge, this is the first report demonstrating the utility of PA-MSHA as an adjuvant to a DNA vaccine. In the in vitro assay, we hypothesized that PA-MSHA would have a stimulatory effect on murine cells NVP-BKM120 944396-07-0 similar to the effects of bacterial flagellin.  Here, we found that PA-MSHA resulted in a decrease in TLR5 level, but significant upregulation of TLR1, TLR2, TLR3, TLR6, TLR7, TLR9, MyD88, and associated adaptor molecules. Among the modifiable risk factors that played a substantial role in previous studies were resting HR, obesity measured by BMI, and BP. Resting HR was the strongest predictor of CAN. Resting HR was considered as an outcome of CAN. If the HR of an individual was more than 90 beats/min, his risk score was at least up to 8. It means the person was a high-risk one, which was consisted with clinical early stage outcome of CAN. Controlling resting HR well might the most important for prevention of CAN complications. In this study, BMI was detected to increase CAN risk at cutoff points suggested for China populations that are lower than those used for people in Western countries. BMI was strong positive correlated with IR and dyslipidemia, and was a strong independent predictor of CAN. In this score system, BMI was an indicator of IR and diabetes status that was the most contributors to CAN. High BP plays a crucial role in progression of CAN. Low HRV and CAN associated with HT. High-risk individuals might benefit from controlling BP to normal status. In general, DM and its duration were considered as two main risk factors for the progression of CAN. In this study, the two factors with high ORs associated with CAN. This suggests that PA-MSHA can successfully trigger TLR pathway activation and upregulation of cytokines and proinflammatory factors independent of TLR5. Followed by their transfer to HEp-2 cells monolayers seeded in DMEM.