Effect of Drug-containing Serum of Clerodendrum bungei on Liver Cancer Cells Through PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

Objective: To investigate the effect of Clerodendrum bungei-containing serum on liver cancer MHCC97-H cells and its possible mechanism from the perspective of phosphatidylinositol 3-kinase(PI3K)/protein kinase (Akt) signaling pathway. Method: The medicinal serum of 15% C. bungei was used to treat MHCC97-H cells. The effect of serum containing C. bungei on cell proliferation was observed by cell counting kit-8(CCK-8) method, in order to select the best time and concentration. The apoptosis was detected by Annexin V-FITC/PI double staining method. Western blot was used to detect the posphatase and tensin homologous gene deleted from chromosome 10 in key proteins (PTEN), phosphoprotein kinase B (p-Akt) and phosphatidylinositol 3-kinase (PI3K)-related protein expression of PI3K/Akt signaling pathway. Real-time PCR was used to detect C. bungei-containing serum on cells for 72 h after activation of nuclear factor-activated B cell kappa light chain(NF-κB) and tumor necrosis factor-α (TNF-α) mRNA expression. Result: The results of CCK-8 showed an inhibitory effect of the C. bungei-containing serum on the proliferation of tumor cells in a dose and time-dependent manner. Among them, the high-dose group had the most obvious inhibitory effect, and the maximum inhibition rates at 24, 48,72 h were 28%, 32%, and 43%, respectively. The results of flow cytometry showed that with the increase of drug-containing serum concentration, the cell growth was observed. The inhibition rate of cells was increased to different degrees, and the inhibition effect was significantly increased in the 72 h intervention group (PC. bungei-containing serum group was 19.48% and 19.72%, compared with the blank group. The difference was significant (PC. bungei-containing serum (PPC. bungei-containing serum could down-regulate the expression of NF-κB and up-regulate the expression of TNF-α mRNA (PConclusion: The medicinal serum of C. bungei can effectively inhibit the proliferation of MHCC97-H hepatoma cells and promote its apoptosis, which may be related to the PI3K/Akt signaling pathway and its key factors.

Detection of peripheral blood Th1/Th2 and Tc1/Tc2 subsets in patients with condyloma acuminatum and its significance / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the role of T-helper (Th) and cytotoxic T (Tc) lymphocyte polarization in the pathogenesis of condyloma acuminatum (CA) and its correlation with recurrence.</p><p><b>METHODS</b>Three-colour immunofluorescent flow cytometry was used to detect the proportion of CD3+ CD8- /IFN-gamma+ (Th1), CD3+ CD8- /IL-4+ (Th2), CD3+ CD8+/IFN-gamma+ (Tc1) and CD3+ CD8+ /IL-4+ (Tc2) cells in the peripheral blood of CA patients and health controls.</p><p><b>RESULTS</b>Compared to health controls, CA patients showed a decreased number of Th1 (P < 0.01) and Tc1 cells (P < 0.05), as well as a decreased Th1/Th2 and Tc1/Tc2 ratio (P < 0.05). Furthermore, in 15 recurrent CA patients the ratio of Th1/Th2 was remarkably decreased (P < 0.01), while the ratio of Tc1/Tc2 had no significant change in comparison with health controls.</p><p><b>CONCLUSION</b>The decrease of Th1 and Tc1 subsets results in relative Th2 and Tc2 predominance, and this tendency is more significant in recurrent CA patients. The Th1 to Th2 and Tc1 to Tc2 shifts in CA patients could be responsible for the fact that human papilloma virus (HPV) is hard to be eliminated.</p>

Determination of Th1/Th2 cytokines and chemokines in patients with allergic diseases and its significance / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine Th1/Th2 cytokines and chemokines in patients with allergic diseases and its clinic significance.</p><p><b>METHODS</b>Serum levels of IFN-gamma, IL-4, IL-5, IL-13, Eotaxin, RANTES and LTB4 were determined from peripheral blood of 64 allergic patients and 21 healthy controls with ELISA.</p><p><b>RESULTS</b>IL-4, IL-5, IL-13 and Eotaxin, LTB4 were increased significantly in serum of allergic patients compared with those of controls (P<0.05). There were no significant differences in serum levels of IFN-gamma and RANTES between patients and controls (P>0.05). Serum levels of IL-4, IL-5, IL-13 and LTB4 were correlated with each other (P<0.01). Eotaxin, RANTES and IFN-gamma levels were also significantly correlated with each other (P<0.05). LTB4 was correlated with Eotaxin as well (P<0.01).</p><p><b>CONCLUSION</b>A wide range of cytokines and chemokines is involved in allergic diseases,which may play their roles in a complex interactive manner.</p>

Effects of drugs known to trigger psoriasis on HaCaT keratinocytes / 药学学报

To investigate whether lithium carbonate, propranolol or chloroquine aggravate psoriasis through influencing cytokines of the psoriatic cytokine network, HaCaT keratinocytes were stimulated with TNF-a after treatment with these drugs. Protein secretion of a set of multiple different cytokines and growth factors in culture supernatants were measured by using a cytokine antibody array technology. Expression of IL-8 and IL-6 mRNA was determined by real-time PCR. In culture supernatants of TNF-alpha-stimulated HaCaT cells, production of IL-6 and TNF-alpha could be enhanced by lithium carbonate; production of IL-6 and a panel of cytokines and growth factors could be enhanced by propranolol hydrochloride; and IL-6 was up-regulated by chloroquine diphosphate as well. Real-time PCR analysis showed a significantly dose-dependent increase of IL-8 and IL-6 mRNA expression in HaCaT cells stimulated with TNF-a as compared to cells without TNF-alpha-stimulation, the mRNA expression of IL-8 was higher than that of IL-6 with the same concentration of TNF-alpha (P < 0.01). Compared with HaCaT cells cultured with medium alone, propranolol hydrochloride at the concentration of 1 x 10(-6) mol x L(-1) could stimulate HaCaT cells to express higher level of IL-6 mRNA (P < 0.05). The drugs investigated show a modulatory effect on certain cytokines and growth factors which are able to modulate inflammatory type of immune reaction present in psoriatic lesions.

Modulatory effect of fumaric acid esters on superoxide-anion generation in human phagocytes / 药学学报

Fumaric acid esters (FAE), mainly dimethylfumarate (DMF), have been shown to be highly efficacious in the treatment of psoriasis. Among the potential side effects of FAE therapy, lymphocytopenia is sometimes observed. In order to address the question whether FAE may interfere with systems of the innate defense, the modulatory role of FAE on the generation of superoxide-anion by human monocytes and neutrophils was studied by measuring the reduction of cytochrome c. Various concentrations of DMF and its metabolite methylhydrogenfumarate (MHF) were used to observe their modulatory effect on superoxide-anion generation by monocytes and neutrophils in response to bacteria (S. aureus and E. coli) and candida (C. albicans). Dexamethasone (DXM, 1 x 10(-7) mol x L(-1)) was also studied at the same time. We found that DXM significantly inhibited superoxide-anion generation from monocytes in response to bacteria and C. albicans, whereas DMF and MHF (10-20 microg x mL(-1)) significantly increased the production of superoxide-anion in monocytes in response to the above mentioned bacteria. DXM, DMF and MHF did not affect superoxide-anion generation of neutrophils. Our data indicate that DMF and MHF enhance superoxide-anion generation in human monocytes as one of the important mechanisms of innate defense against microorganisms.

HPV DNA vaccines expressing recombinant CRT/HPV6bE7 fusion protein inhibit tumor growth and angiogenic activity / 病毒学报

This paper was to study the angiogenic inhibitory effect and the potential antitumor effect of the constructed recombinant DNA vaccine CRT/HPV6bE7 in vivo. The C57BL/6 mice were vaccinated respectively with recombinant CRT/HPV6bE7 DNA plamids. The inhibitory effects on angiogenesis of generated vaccines in vivo were evaluated by a bFGF-induced angiogenesis assay using the Matrigel kit. To investigate the potential antitumor effect, the mean tumor weights, sizes and tumor appearing times were measured in C57BL/6 mice treated with HPV6bE7-expressing B16 cells. The results indicated that the recombinants CRT180/HPV6bE7 and CRT180 showed strong anti-angiogenic effects in bFGF-induced angiogenesis in vivo. Moreover, CRT180/HPV6bE7 and CRT180 DNA vaccines could significantly inhibit the tumor growth in tumor challenge experiment, and CRT180/HPV6bE7 was superior to other vaccines in delaying tumor formation time, limiting tumor size and weight in tumor protection experiment. In conclusion, recombinant CRT180/HPV6bE7 DNA could elicit a most efficient anti-angiogenic effect and inhibit tumor growth in mice inoculated with DNA vaccines. The antiangiogenic activity of CRT were suggested residing in a domain between CRT 120-180 aa.

1 alpha, 25-dihydroxyvitamin D3 and its analogues modulate the phagocytosis of human monocyte-derived dendritic cells / 药学学报

<p><b>AIM</b>To investigate the role of 1 alpha, 25-dihydroxyvitamin D3 (calcitriol) and its analogues tacalcitol and 24, 25(OH)2D3 on the phagocytosis of human monocyte-derived dendritic cells (MoDC).</p><p><b>METHODS</b>MoDC were generated in vitro by differentiating monocytes in the presence of GM-CSF and IL-4 for 5 days. Expression of mannose receptor (MR) and Fc gamma receptors (Fc gamma Rs) by MoDC was analysed by flow cytometry. Zymosan ingestion was measured to assess the phagocytosis of MoDC.</p><p><b>RESULTS</b>MoDC expressed high level of MR and Fc gamma Rs and showed the capacity of zymosan ingestion. Calcitriol and tacalcitol but no 24, 25(OH)2D3 not only upregulated the expression of MR and Fc gamma Rs on MoDC but also correspondingly enhanced their phagocytosis by increasing zymoasan ingestion. Furthermore, the upregulatory role occurred in the early stage of MoDC differentiation and was irreversible. The upregulatory role of calcitriol was dose dependent.</p><p><b>CONCLUSION</b>Calcitriol and its analogue tacalcitol may play an important role in dendritic cell binding and capturing foreign antigens at the initiation of immune response.</p>

Effects of fumaric acid esters on differentiation of dendritic cells in vitro / 浙江大学学报·医学版

OBJECTIVE: To investigate the effect of anti-psoriatic drug fumaric acid esters (FAE) on the differentiation of dendritic cells (DC). METHODS: Dendritic cells were obtained by differentiating human monocytes in vitro. Flow cytometry was used to analyse the effect of FAE on cell surface expression of CD1a, CD14, CD40, CD80, CD86 and HLA-DR by monocyte-derived dendritic cells (MoDC). Mixed lymphocyte reaction (MLR) was made to demonstrate the influence of FAE on T cell stimulatory activity of MoDC. RESULTS: Dimethylfumarate and methylhydrogenfumarate-calcium-salt (0.01 approximate, equals 100 mg/L) inhibited MoDC differentiation as well as reducing the capacity of MoDC to stimulate lymphocytic proliferation in MLR. CONCLUSION: The mode of action of FAE in pso riasis may be mediated by inhibition of DC differentiation.