Tollip suppresses MyD88-mediated NF-κB activation by enhancing MyD88 ubiquitination levels in large yellow croaker (Larimichthys crocea).

Toll-interacting protein (Tollip) plays an important role in the innate immune response by negative regulation of the TLR-IL-1R signaling pathway. MyD88 serves as a universal adaptor in TLR-mediated NF-κB activation. However, the regulation mechanisms of Tollip in piscine MyD88-mediated NF-κB activation is largely unknown. In the present study, the cDNA sequence of LcTollip was identified from the large yellow croaker (Larimichthys crocea). The putative LcTollip protein encoded 275 amino acid residues, containing a N-terminal TBD domain, a central C2 domain, and a C-terminal CUE domain. Quantitative PCR showed that the most predominant constitutive expression of LcTollip was detected in spleen. In addition, LcTollip transcripts enhanced significantly after LPS and poly I:C challenge (P < 0.05). Cellular localization revealed that LcTollip existed in the cytoplasm and nucleus. Furthermore, the overexpression plasmids of wild type LcTollip as well as its six domain truncated mutants of LcTollip were constructed by overlap PCR. Dual luciferase analysis showed that NF-κB activation could not be induced by overexpression of LcTollip or its domain truncated mutants alone. However, the LcMyD88-induced-NF-κB activation was significantly suppressed by overexpression with LcTollip, and the truncated mutants LcTollip-ΔTBD, LcTollip-ΔC2, LcTollip-ΔCUE and LcTollip-ΔTBDΔCUE while not by LcTollip-ΔLR and LcTollip-ΔTBDΔC2. Moreover, co-immunoprecipitation (Co-IP) assay revealed that the interaction between LcTollip and LcMyD88 was through CUE domain. More interesting, IP and immunoblotting examination of HEK293T cells co-transfected with LcMyD88, LcTollip and HA-ubiquitin showed that LcMyD88 induced a dose-dependent de-ubiquitination of LcTollip while LcTollip enhanced a dose-dependent ubiquitination of LcMyD88. However, protein degradation investigation displayed that the proteolysis and ubiquitination of LcMyD88 were not connected. Our findings suggested that the LcTollip might involve in negative regulation TLR pathway by suppressing LcMyD88-mediated immune activation and improving the ubiquitination level of LcMyD88.

[Effects of Compound Aconitum Polysaccharide on Glycosylation of Tumor Cells].

Objective To observe inducing or inhibiting effects of Chinese medicine (CM) poly- saccharides on glycoprotein chain synthetized different glycosyltransferases, thus disclosing targets of CM polysaccharides and its mechanisms. Methods In vivo anti-tumor effects of CM polysaccharides were observed using the inhibiting rate of tumor growth by dividing different Aconitum containing groups. Effects of CM polysaccharides on liver cancer cell SK-HEP-1 glycosyltransferase and tumor related gene expressions were observed. Meanwhile, changes of polylactosamine expression were detected using flow cytometry (FCM) with polylactosamine specific biotin labeling lectin. Results Compared with the model group, the average tumor weight was significantly lower in each medication group (P <0. 01). Compared with the adriamycin group, no significant difference in average tumor weight of the three compound groups (P>0. 05). The expression level of polylactosamine was reduced after adding Aconitum polysac- charide; and CM compound polysaccharides respectively. Conclusions Polysaccharide compound showed similar anti-tumor effect as that of adriamycin. Besides, polylactosamine expression level was reduced in the three compound groups along with increased prepared Aconitum polysaccharide, with more obvious anti-tumor effects shown.

Efficacy of short-term cordyceps sinensis for prevention of contrast-induced nephropathy in patients with acute coronary syndrome undergoing elective percutaneous coronary intervention.

Contrast-induced nephropathy (CIN) is one of the major causes of hospital-acquired acute renal failure. The pathophysiological mechanism of CIN remains unknown. There has been little evidence regarding the effects of Traditional Chinese Medicine (TCM) on CIN. Cordyceps sinensis (CS), a traditional Chinese herb, has been widely used clinically for the prevention of the progression of renal failure. We performed a prospective, randomized controlled trial to investigate the role of CS in the prevention of CIN in patients with acute coronary syndrome (ACS) undergoing elective percutaneous coronary intervention (PCI). The 150 ACS patients were randomly assigned to three groups, basic treatment group (n=51), standard CS therapy group (n=49, corbrin capsule 2 g, 3 times/d were used 3 days before and after angiography), and intensive CS therapy group (n=50, corbrin capsule 3 g, 3 times/d were used 3 days before and after angiography). Renal function was assessed at the time of hospital admission and on days 1, 2, and 3 after PCI. CIN occurred in 13 of 150 patients (8.67%). The incidence of CIN was lower in the CS treatment groups than in the basic treatment group (P<0.05), and a significant decrease in the incidence of CIN in the intensive CS therapy group was shown (P<0.01). In conclusion, prophylactic treatment with CS during the peri-procedural stage in ACS patients undergoing elective PCI has a preventive role against CIN, and intensive CS therapy could be more effective.

[Effects of supplementing qi and activating blood circulation method on platelet aggregation rate, adhesion rate and thromboxane B2 level in patients with stable angina pectoris and intolerable to aspirin].

OBJECTIVE: To investigate the effects of supplementing qi and activating blood circulation method (YQHX) on platelet aggregation rate (PAgR), platelet adhesion rate (PAdR) and thromboxane B2(TXB2) level in patients with stable angina pectoris and intolerable to aspirin. METHODS: Seventy-six out-patients with stable angina (qi deficiency and blood stasis syndrom) pectoris intolerable to aspirin were randomized into two groups, 40 in the treated group and 36 in the control group. Both received conventional Western medicinal treatment with YQHX to the treated group additionally, for 1 month. PAgR, PAdR, TXB2 level, platelet count, hemoglobin concentration and fecal occult blood were measured before and 1 month after treatment, and the cardiac events as well as diges tive symptoms occurred in the observation period were recorded. RESULTS: PAgR, PAdR and TXB2 level lowered in the treated group after 1-month treatment showed a significant difference to those of baseline, and also to those in the control group (all P <0.01). But no significant difference was found between pre-treatment and post-treatment, also between the two groups in platelet count, hemoglobin concentration, fecal figure and incidence of adverse cardiac events, as well as digestive symptoms (P > 0.05). CONCLUSION: YQHX can effectively inhibit the platelet function in patients with stable angina pectoris without aggravation of digestive symptoms. Cardiac event reducing effect of YQHX was not seen in this study, it is necessary for large sampled study for confirmation.

[Experimental study on Qi-replenishing and Yin-nourishing in inhibiting tumor metastasis].

OBJECTIVE: To study the effect of Qi-replenishing and Yin-nourishing in inhibiting tumor metastasis. METHODS: Fifty mice were randomly divided into replenishing Qi group, replenishing Yin group, replenishing Qi and Yin group, chemotherapy group and control group. Models of Lewis lung cancer and B(16) melanoma were observed in vivo. The tumor weight inhibiting rate, metastatic lung tumor, concentrations of t-PA and TXB(2) in tumor tissue and IL-2 and TNF-alpha in spleen were observed. RESULTS: The tumor weight inhibiting rates of replenishing Qi and Yin group for Lewis lung cancer and B16 melanoma were both 42.9%. Replenishing Qi and Yin could decrease the contents of TXB(2) and 6-keto-PGF(1alpha), and increase the levels of IL-2 and TNF-alpha. CONCLUSION: Replenishing Qi and Yin can promote the healthy energy to inhibiting tumor metastasis.