Objective To further investigate the effect of sinomenine (SIN) on TNF-α-induced VCAM-1 expression in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were isolated from freshly collected umbilical cords. Positive control samples were stimulated with TNF-α, but free of SIN. Negative control samples were treated in the same way, but without TNF-α and SIN. Experimental samples were co-cultured with TNF-α and SIN at various concentrations (0.25, 0.5, and 1.0 mol/L), or TNF-α and dexamethasone (Dex) at concentration of 1.0×10-6 mol/L, or TNF-α with Dex (at concentration of 1.0×10-6mol/L) and SIN at different concentrations (0,25, 0.5, and 1.0 mmol/L) (co-treated groups). VCAM-1 expression was detected by flow cytometry (FCM). Results SIN inhibited expression of VCAM-1 in TNF-α-induced HUVECs, the best effect was shown in the 1.0 mmol/L SIN treated group. VCAM-1 decreased more markedly in the co-treated groups. Conclusion SIN inhibits TNF-α-induced VCAM-1 expression on HUVECs in vitro, and SIN maybe synergistic with Dex in inhibiting TNF-α-induced VCAM-1 expression on HUVECs in vitro.

BACKGROUND: Apolipoprotein(a) [Apo(a)] plays some role in promoting the formation of atherosclerotic plaque, and contains pentanucleotide repeats(PNR), which has a key value in genic research and in forecast on the increased risk of early atherosclerosis cerebral infarction (ACI). But the relationship between ACI and Apo(a) PNR in different races needs to be further investigated.OBJECTIVE: To investigate the relationship between the Apo(a) PNR polymorphism and ACI.DESIGN: A case-control study based on the ACI patients and normal people of Han nationality in Hubei.SETTING: Department of Laboratory in a hospital of a university.PARTICIPANTS: From February 1998 to March 1999, 82 ACI patients (ACI group) and 153 healthy controls(control group) were selected from the Department of Neurology, Central South Hospital and Yatai Hospital of Wuhan University. All patients were Han nationality in Hubei without any relatives.METHODS: Serum lipoprotein(a) [Lp(a) ], total cholesterol(TC), high density lipoprotein cholesterol(HDL-c), low density lipoprotein cholesterol (LDL-c), triglyceride (TG), apolipoprotein A Ⅰ (ApoA Ⅰ) and apolipoprotein B (Apo B) were tested respectively. Meanwhile, the PNR in the 5' control region of the Apo(a) was detected with polymerase chain reaction(PCR) and high voltage polyacrylamide gels electrophoresis. The results were analyzed with controlled analysis.RESULTS: The levels of serum Lp(a) [ (239.9 ±225.4) mg/L], TC [(4.76±0.74) nmol/L], TG[(1.74±0.60) mmol/L] and LDL-C [ (2.84 ± 0. 63) mmol/L] were remarkably higher in ACI group than those in control group, which were(133.5 ±97.7) mg/L in serum Lp(a), (4. 29±0.72) mmol/L in TC, (1.05±0.52) mmol/L in TG and(2.84±0.63) mmol/L in LDL-C, however, the level of HDL-C[ (0.88± 0.17) mmol/L] was remarkably lower in ACI group than that in control group [ ( 1.03 ± 0. 35 ) mmol/L], the differences were all significant( t = 3.65to9.18, P ＜ 0.01) . The levels of ApoA Ⅰ [(1.13±0.15) mmol/L,(1.25±0.19) mmol/L] and ApoB[(0.93±0.12) mmol/L, (0.89± 0. 15 ) mmol/L] were no significant difference compared with those in control group. The duplicated frequency of the allele(TTTTA) 5(0. 098) in the ACI was remarkably higher than that in control(0. 026) (x2 = 5.62, P＜ 0. 05), The frequency of the allele(TTTTA) 9 in the ACI(0. 073) was remarkably lower than that in control (0. 213 ) (x2 = 7.83, P ＜ 0.01 ), The frequency of the allele(TTTTA) 5 was also associated with low TC and high Lp(a) levels.CONCLUSION: It is suggested that the Apo(a) PNR polymorphism are associated with the susceptibility to ACI, and involved in the development of ACI.

Objective To study the effect of mitogen-activated protein kinas/extracellular signalregulated kinase (MAPK/ERK) signaling pathway on cell proliferation modulated by Sonic Hedgehog (Shh) signaling in fibroblast-like synoviocytes (FLS) isolated from patients with active rheumatoid arthritis (RA).Methods The synovial tissue were collected by the synovial arthroscopic debridement or arthroscopic synovectomy of RA patients with active disease activity [disease activity score(DAS)28 ≥3.2].The RA-FLS were primarily cultured by the explanted culture,and then were treated with Shh agonist purmorphamine,inhibitor cyclopamine or MAPK/ERK signaling pathway inhibitor U0126,respectively.Western blotting was used to examine the phosphorylation level of ERK 1/2 (p-ERK1/2),which was the critical protein of MAPK/ERK signaling.The cell proliferation activity was detected using cell proliferation and cytotoxicity kit-8 (CCK8),and the cell proliferation rate was detected using a flow cytometry.Analysis of variance and Kruskal-Wallis H(K) test were used for statistical analysis.Results Compared with the control group,purmorphamine transiently increased p-ERK1/2 protein at the concentration of 1 μmol/L,and the peak activations of p-ERK1/2 took place at 15 min (P＜0.01).Cyclopamine and U0126 decreased the expression ofp-ERK1/2 protein (P＜0.01).After the RA-FLS treated with purmorphmine(1 μmol/L)for 48 hours,the cell proliferation activity was (114±4)％ and the percentage of S phase cells was (8.39±0.60)％,which was significantly higher than those of the control group (100±0)％ (P＜0.01) and (3.29±0.69)％ (P＜0.01).After treated with cyclopamine (10 μmol/L) for 48 hours,the cell proliferation activity of RA-FLS was (89±1)％ (P＜0.05) and the percentage of S phase cells was (1.53±0.22)％ (P＜0.05).When co-treated with purmorphamine (1 μmol/L) and U0126 (10 μmol/L),the cell proliferative activity was (89±2)％ (P＜0.05) and the percentage of S phase cells was(1.07±0.25)％(P＜ 0.05).Conclusion Shh may promote proliferation of RA-FLS via modulating MAPK/ERK signaling,which in turn contributes to hyperplasia of synovium and ultimately leading to RA.

Objective To establish a serological detection method for EA-D-IgA antibody,and to evaluate its diagnostic efficacy for nasopharyngeal carcinoma in different clinical stage.Methods EA-D-IgA antibody serological detection method was established by using the polypropylene microplate with eukaryotic expression product of BMRF1 whole gene fragment of EB virus.Fifteen early stage (stage Ⅰ and Ⅱ) and 48 advanced (stage Ⅲ and Ⅳ) patients with nasopharyngeal carcinoma in Shanxi Provincial Cancer Hospital and Shanxi Dayi Hospital from April 2012 to August 2017,and serum samples from 40 patients with rhinitis who were treated at Shanxi Dayi Hospital from October 2016 to October 2017 were examined respectively by using the constructed EA-D-IgA antibody detection method.The positive detection rate of EA-D-IgA antibodies in different groups was calculated.When the patients with rhinitis were used as the differential control,the diagnostic efficacy of this index for different stages of nasopharyngeal carcinoma was evaluated.Results EA-D-IgA antibody serological method was successfully established.The positive detection rate of EA-D-IgA antibody in early nasopharyngeal carcinoma,advanced nasopharyngeal carcinoma and rhinitis control was 60.0 ％ (10/15),68.3 ％ (33/48) and 5.0 ％ (2/40) respectively.The differences between early stage nasopharyngeal carcinoma and the rhinitis control,advanced nasopharyngeal carcinoma and the rhinitis control were statistically significant (x2 =20.625,P =0.000;x2 =37.017,P =0.000).The difference between early nasopharyngeal carcinoma and advanced nasopharyngeal carcinoma was not statistically significant (x2 =0.394,P =0.530).When compared with the patients with rhinitis,the diagnostic sensitivity,specificity,positive predictive value,negative predictive value was 60.0 ％ and 68.3 ％,95.0 ％ and 95.0 ％,81.8 ％ and 94.3 ％,86.4 ％ and 71.7 ％ respectively in early nasopharyngeal carcinoma and advanced nasopharyngeal carcinoma.Conclusion The method constructed in this study effectively improves the efficacy of EA-D-IgA antibody detection in serological diagnosis of nasopharyngeal carcinoma,which can be used as an adjunct for early diagnosis of nasopharyngeal carcinoma,yet not as a reference for clinical staging.