Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-κB (TLR/NF-κB) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells / 中国实验方剂学杂志

Objective::To investigate the effect of total saponin of Dioscoreae Collettii Rhizoma (TSD) on Toll-like receptor/nuclear factor-κB (TLR/NF-κB) signaling pathway induced by monosodium urate in THP-1 cells, in order to explore the possible mechanism of anti-gout arthritis. Method::Phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cells were differentiated into macrophages, divided into normal group, model group, low, medium and high-concentration TSD groups (1, 3, 10 mg·L-1) and colchicine group (0.2 mg·L-1). Except the normal group, the other groups were stimulated with 400 mg·L-1 monosodium urate to replicate an inflammation model in vitro. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay, the levels of inflammatory factors tumor necrosis factor-α(TNF-α ) and interleukin-1β(IL-1β) were detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and NF-κB were detected by Western blot. The mRNA levels of TLR4, NF-κB and Pro-IL-1β were measured by real-time fluorescence quantitative PCR (Real-time PCR), and the nuclear shift of NF-κB p65 was detected by immunofluorescence. Result::0~32 mg·L-1 TSD has no effect on cell viability. Compared with the normal group, the secretion levels of inflammatory factors TNF-α and IL-1β in the model group were significantly increased (P<0.01), and the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were increased (P<0.01). Compared with the model group, 1-30 mg·L-1 TSD significantly down-regulated the secretion of inflammatory factors TNF-α and IL-1β (P<0.01), the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were decreased (P<0.05, P<0.01), and the NF-κB p65 partially trans-located to the cytosol and the superposition in the nucleus were decreased, inhibiting the nuclear translocation of NF-κB p65. Conclusion::TSD may exert an anti-inflammatory effect by down-regulating the expressions of TLR4, NF-κB and Pro-IL-1β mRNA and reducing the secretion of inflammatory factors TNF-α and IL-1β.

Regulatory effect of Di'ao Xinxuekang on TLR4/MyD88/NF-κB signaling pathway in atherosclerotic rats / 中国中药杂志

The aim of this paper was to observe the effect of Di'ao Xinxuekang(DXXK) on TLR4/MyD88/NF-κB signaling pathway in atherosclerotic rats, and to explore its anti-atherosclerotic mechanism. Sixty SD rats were randomly divided into normal group, model group, atorvastatin group(4.0 mg·kg~(-1)), and DXXK groups(100, 30, 10 mg·kg~(-1)), with 10 rats in each group. The atherosclerosis model was induced by high fat diet plus vitamin D_2. Experimental drugs were administered intragastrically once daily for 8 weeks starting from the 9 th week. Biochemical analyzers were used to detect levels of triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) in blood lipid. The levels of serum tumor necrosis factor(TNF)-α, interleukin(IL)-6 and IL-1β were detected by ELISA. Pathological changes of aortic tissues were observed by using Sudan Ⅳ and HE staining. The mRNA and protein expressions of TLR4, MyD88 and NF-κB p65 in aortic tissues were detected by RT-PCR and Western blot, respectively. As compared with the model group, TC, TG, and LDL-C levels in serum were significantly decreased, HDL-C content was significantly increased, and levels of TNF-α, IL-6, and IL-1β in serum were significantly decreased in atorvastatin group and DXXK high and middle dose groups. Aortic lesions in atorvastatin group and DXXK group were significantly improved, and the mRNA and protein expressions of TLR4, MyD88, NF-κB p65 in the aorta were decreased. DXXK has a preventive and therapeutic effect on atherosclerosis in rats, and its mechanism may be related to inhibiting inflammatory reaction by regulating TLR4/MyD88/NF-κB signal transduction, thereby inhibiting the progression of atherosclerosis.

[Efficiency of Inducing CIK from Cryopreserved PBMNC by Using Immune Cell SR].

OBJECTIVE: To investigate the efficiency of inducing CIK from peripheral blood mononuclear cells(PBMNC) by using immune cell serum replacement(immune cell SR), so as to provide a new strategy for the industrialized production of immune cells. METHODS: The PBMNC of healthy volunteers were collected, and these cells were thawed after short-term cryopreservation and cultured to induce CIK cells. The cells viability was measured by trypan blue exclusion, the phenotypes were analyzed by flow cytometry, and the cytotoxicity was determined by Calcein-AM/PI double staining. RESULTS: In cryopreserved PBMNC, the control group cells failed to normally proliferate. Cell proliferation ratio was low in 2% SR group in comparison with the fresh group, and the difference was significant (P<0.05), however, differences were not statistically significant between 5% SR and fresh group or between 10% AP and fresh group. CD3+, CD3+CD8+ and CD3+CD56+ cell subsets were not significantly different before and after cryopreservation (P>0.05). After being cultured, CD3+, CD3+CD4+, CD3+CD8+, CD3+CD56+ and CD3-CD56+ subsets and the cytotoxicity in vitro were not significantly different among all group(P>0.05). CONCLUSION: 5% SR without the protein of animal origin can be safely used as a substitute for autologous plasma in CIK induced from cryopreserved PBMNC by culture, thus providing a basis for the application of cryopreservation technique of immune cells to cell therapy.

Construction and development of a training system for military pharmacy professionals / 军事医学

Amid continuous development of new military revolutions,the cultivation of high-quality professionals practical ability is becoming increasingly important.By learning from the successful experience and practices from foreign systems for training practical skills of graduate students majoring in military pharmacy,the education mode and methods for training military pharmacy postgraduates were discussed in this paper in order to optimize the training mode of practical skills for students of military pharmacy,explore a suitable training mode in China,improve teaching methods and formulate a set of scientific and optimized strategies.It is hoped that the practical ability of military pharmacy graduate students will be improved considerably.