The Blockade of Mitogen-Activated Protein Kinase 14 Activation by Marine Natural Product Crassolide Triggers ICD in Tumor Cells and Stimulates Anti-Tumor Immunity.

Immunogenic cell death (ICD) refers to a type of cell death that stimulates immune responses. It is characterized by the surface exposure of damage-associated molecular patterns (DAMPs), which can facilitate the uptake of antigens by dendritic cells (DCs) and stimulate DC activation, resulting in T cell immunity. The activation of immune responses through ICD has been proposed as a promising approach for cancer immunotherapy. The marine natural product crassolide, a cembranolide isolated from the Formosan soft coral Lobophytum michaelae, has been shown to have cytotoxic effects on cancer cells. In this study, we investigated the effects of crassolide on the induction of ICD, the expression of immune checkpoint molecules and cell adhesion molecules, as well as tumor growth in a murine 4T1 mammary carcinoma model. Immunofluorescence staining for DAMP ectolocalization, Western blotting for protein expression and Z'-LYTE kinase assay for kinase activity were performed. The results showed that crassolide significantly increased ICD and slightly decreased the expression level of CD24 on the surface of murine mammary carcinoma cells. An orthotopic tumor engraftment of 4T1 carcinoma cells indicated that crassolide-treated tumor cell lysates stimulate anti-tumor immunity against tumor growth. Crassolide was also found to be a blocker of mitogen-activated protein kinase 14 activation. This study highlights the immunotherapeutic effects of crassolide on the activation of anticancer immune responses and suggests the potential clinical use of crassolide as a novel treatment for breast cancer.

A gender-specific COMT haplotype contributes to risk modulation rather than disease severity of major depressive disorder in a Chinese population.

BACKGROUND: COMT rs4680 Val158 allele is associated with high MB-COMT protein expression and elevated activity compared to the Met158 allele in post-mortem brains. A meta-analysis study suggested the link between COMT SNPs and MDD risk; in addition, MB membrane-bound (MB-COMT) specific genetic variation was reported that influences predisposition to depression amongst females. METHODS: Four tagSNPs, including rs4680, were genotyped. 268 MDD subjects and 223 controls were enrolled. MDD severity was rated by HDRS. Total-COMT and MB-COMT mRNA were detected by quantitative PCR. COMT protein and activity were assayed by western blot and methyltransferase assay, respectively. RESULTS: Haplotype TG of rs4633-rs4680, rs4646312 C, and rs4633 T allele might be linked to MDD vulnerability. Haplotype TG may interact with gender and affect MDD risk, since female haplotype TG carriers were estimated for a 9.17-fold higher risk than counterparts. COMT SNPs were not associated with HDRS scores. Haplotype TG female controls had higher MB-COMT protein, whereas non-TG female controls had higher soluble cytoplasmic (S-COMT) protein than other groups. COMT activity was much higher in controls than in MDD subjects. LIMITATIONS: Restricted numbers of homozygous TG carriers were recruited and analyzed for COMT mRNA, protein and activity. Only peripheral blood samples were used. CONCLUSIONS: A female-specific haplotype (haplotype TG)-MDD vulnerability association was found. TG female controls had higher MB-COMT protein and S-COMT. Altogether, high COMT protein and activity in female TG controls may be predisposing factors for enhanced MDD risk, though not correlated to MDD severity as rated by HDRS.

Effects of Scutellariae Radix on gene expression in HEK 293 cells using cDNA microarray.

The aim of the present study was to elucidate the molecular mechanisms underlying the anti-inflammatory effect of Scutellaria Radix (SR). The complementary DNA (cDNA) microarray method was used to survey the effects of SR on the changes of gene expression profile in HEK293 cells. Based on differential expression, 66 genes were selected for further analysis from 9,600 candidate genes in the microarray; 23 genes were validated by RT-PCR. The broad spectrum of the differentially expressed genes, including those associated with inflammation, immune response, energy metabolism, as well as others, such as ISGF3G, IL6ST, CD98, ATP5G2, PHKG2, YB-1 and SLC7A4, indicate overall cellular response to SR treatment. Our results suggest that the anti-inflammatory effect of SR may be related to IL6ST down-expression, and over-expression of CD98. Moreover, SR-related improvement in immune response may be related to the ISGF3G over-expression.

Differential gene expression in hemodialysis patients with "cold" zheng.

The aim of the present study was to search for the differential gene expression and measure the serum level of a number of biochemical parameters in the cold zheng (CZ) and non-cold zheng (NCZ) in patients receiving hemodialysis. Hemodialysis (HD) patients were randomly selected from the CZ and NCZ groups. The between-group differences in gene expression were assessed using complementary DNA (cDNA) microarray. Differential gene expression was further validated by real-time reverse transcriptase polymerase chain reaction (RT-PCR). Our results demonstrated that the up-regulation of the inflammation-associated genes, ALOX5AP, S100A8 and S100A12, down-regulation of the genes related to immunity (DEFA4), metabolism (GNG11, PYGB, PRKAR2B), and growth/proliferation (HSF2, DDR2, TK1) were found in the CZ group. Furthermore, the CZ HD patients had significantly lower serum albumin levels compared with their NCZ counterparts (3.31 +/- 0.08 g/dL versus 4.18 +/- 0.12 g/dL). It appears reasonable to conclude that up-regulated inflammatory-gene expression (ALOX5AP, S100A8 and S100A12) may play an important role in CZ HD patients.

Anxiolytic effect of berberine on exploratory activity of the mouse in two experimental anxiety models: interaction with drugs acting at 5-HT receptors.

The aim of this study was to assess the anxiolytic effect of berberine (abbrev. BER) using two experimental anxiety models in the mouse. In the black and white test of anxiety, berberine (100, 500 mg/kg) produced an increase in the first time entry, time spent in the white section, and total changes between two compartments. On the other hand, in the elevated plus-maze test, berberine (100, 500 mg/kg) produced an increase in the time spent and arm entries in the open arms, and a decrease in the time spent and arm entries in the closed arms. Berberine (500 mg/kg) decreased locomotor activity in mice. Furthermore, BER at 100, 500 mg/kg decreased concentrations of NE, DA and 5-HT, and increased the concentrations of VMA, HVA and 5-HIAA in the brain stem. BER also attenuated the anxiogenic effect of WAY-100635, 8-OH DPAT and DOI and enhanced the anxiolytic effect of BUS, p-MPPI and RIT in the elevated plus-maze. These results suggested that berberine at 100 mg/kg had a significant anxiolytic-like effect, which was similar to that observed with 1 mg/kg diazepam and 2 mg/kg buspirone. The anxiolytic mechanism of BER might be related to the increase in turnover rates of monoamines in the brain stem and decreased serotonergic system activity. Moreover, BER decreased serotonergic system activity via activation of somatodendritic 5-HT1A autoreceptors and inhibition of postsynaptic 5-HT1A and 5-HT2 receptors.

Osthole improves aspects of spatial performance in ovariectomized rats.

The present study was designed to investigate the ameliorating effects of Cnidiuim monnieri L. Cusson (CM) and osthole, a constituent of CM, on the spatial performance deficit in scopolamine (SCOP)-treated or ovariectomized (OVA) rats. CM improved the deficit of spatial performance, and reversed the lower plasma estradiol levels caused by SCOP in female rats. In addition, osthole (3 and 10 mg/kg, s.c.) improved the performance deficit in OVA rats. It (10 and 30 micrograms/brain, icv) also improved the performance deficit caused by SCOP in intact female rats, and at 30 micrograms/brain improved the deficit in OVA rats. However, osthole did not alter the latency swum to reach the visible target in SCOP-treated and OVA rats. Accordingly, we suggested that osthole is an active constituent of CM, and possesses ameliorating effects on the spatial performance deficits in SCOP-treated female rats or OVA rats. The action mechanism of the effects of osthole on performance deficits was related to the estrogen-like properties and activating the central cholinergic neuronal system.