SOX18 meditates the resistance of Bmi1-expressing cells to cetuximab in HNSCC.

OBJECTIVE: Head and neck squamous cell carcinoma (HNSCC) is the most common type of malignancy in the head and neck region worldwide. The therapeutic strategies for HNSCC remain unsatisfying and limited. Here, we found a population of resistant Bmi1-expressing cells in the presence of cetuximab treatment and reported a novel role of SRY-box transcription factor 18 (SOX18), a member of the SOX family, in promoting HNSCC resistance to cetuximab. This study aimed to investigate the regulatory mechanism of Sox18 in Bmi1-positive cells and to search for better therapeutic targets. METHODS: We successfully obtained Bmi1CreER , RosatdTomato , and RosaDTA mice and identified Bmi1-expressing cells through lineage tracing. SOX18 expression in HNSCC and normal tissues was analyzed by immunohistochemistry, colocalization of Sox18, and Bmi1-expressing cells was analyzed by immunofluorescence, and SOX18 expression in SCC9 cell lines was quantified by western blotting and quantitative real-time PCR. The investigation of the mechanism of SOX18-mediated cetuximab resistance in Bmi1-positive cells was based on the analysis of single-cell RNA-seq data obtained from the Gene Expression Omnibus (GEO) database. Western blotting was performed to verify the results obtained from the single-cell RNA-seq analysis. RESULTS: In our study, we demonstrated that Bmi1-expressing cells were resistant to cetuximab treatment and that depletion of Bmi1-expressing cells improved cetuximab efficacy in HNSCC. We then discovered that Sox18 mediated the stem cell-like properties of Bmi1-expressing cells and promoted cellular cetuximab resistance through an oxidative phosphorylation pathway. There was a significant downregulation of key genes in the oxidative phosphorylation pathway in Sox18 knockout cell lines. CONCLUSIONS: Taken together, the findings of our study suggest that Sox18 mediates the resistance of Bmi1-expressing cells to cetuximab in HNSCC via the oxidative phosphorylation pathway.

Fucoidan Protects Against High-Fat Diet-Induced Obesity and Modulates Gut Microbiota in Institute of Cancer Research Mice.

Fucoidan possesses various biological activities, such as anticoagulant, immunomodulatory, anti-inflammatory, potential antioxidant, and others. In this study, we investigated the effect of fucoidan on high-fat diet-induced obesity, inflammation, and gut microbiota in Institute of Cancer Research mice. Mice were gavaged with 50 mg/(kg·d) (Fuc0.5 group) or 250 mg/(kg·d) (Fuc2.5 group) of fucoidan for 5 weeks. Fucoidan alleviated obesity and tissue damage by decreasing body weight and body mass index, decreasing body weight gain, improved organ index, liver steatosis, and improved the structure of the small intestine. In addition, fucoidan decreased total cholesterol, triglyceride, and low-density lipoprotein cholesterol, and increased high-density lipoprotein cholesterol. Moreover, fucoidan reduced serum lipopolysaccharide concentrations, tumor necrosis factor-α, and total bile acid. Furthermore, fucoidan improved the structure of gut microbiota and significantly increased the abundance (Shannon diversity index, evenness, and Faecalibacterium prausnitzii) determined by denaturing gradient gel electrophoresis and quantitative PCR. In conclusion, our study provides a scientific basis for fucoidan as a functional food for modulating the gut microbiota and protecting against obesity.

Probiotic fermentation of Ganoderma lucidum fruiting body extracts promoted its immunostimulatory activity in mice with dexamethasone-induced immunosuppression.

Ganoderma lucidum is a legendary traditional Chinese medicine with various bioactivities. This study was conducted (a) to explore the in vitro fermentation of the water extracts of G. lucidum fruiting body with Lactobacillus acidophilus and Bifidobacterium breve and (b) to investigate the effect of fermentation broth (GLFB) on dexamethasone (DEX)-induced immunosuppressed mice. Our results demonstrated that probiotic fermentation of G. lucidum fruiting body extracts underwent structural changing of major ganoderic acid components, such as ganoderic acid A (GA) into GC2, and this fermentation process involves changing of several metabolic pathways in the probiotic strains. GLFB could significantly improve the immunity, intestinal integrity, and gut microbiota dysbiosis in DEX-treated mice, and the immunostimulatory activity of GLFB was found closely related to its direct regulation on the expansion of CD4+ T cells in Peyer's patches of mice. These data implied that probiotic fermentation of G. lucidum fruiting body extracts promoted its immunostimulatory activity via biotransformation of components such as GA. This research provides a theoretical support for the development and application of G. lucidum fermentation by probiotics.

Fucoidan and galactooligosaccharides ameliorate high-fat diet-induced dyslipidemia in rats by modulating the gut microbiota and bile acid metabolism.

OBJECTIVES: Dyslipidemia is an important risk factor for cardiovascular diseases. Fucoidan (FUC) is a polysaccharide extracted from brown marine algae with various biological activities. Galactooligosaccharides (GOS) are important prebiotics that exert benefits on the intestinal microbiota. The aim of this study was to investigate the effects of FUC and GOS on dyslipidemia in rats by modulating the gut microbiota and bile acid metabolism. METHODS: Twenty-four male inbred Sprague-Dawley (SD) rats aged 8 wk were fed a normal or high-fat diet (HFD) for 8 wk. During the feeding period, rats were gavaged with normal saline solution, FUC solution (100 mg/kg),or GOS solution (800 mg/kg), or a combination of both once daily. Serum biochemical parameters were determined, and the gut microbiota were analyzed via 16S rRNA gene sequencing. Bile salt hydrolase (BSH) activity in the small intestinal contents was also analyzed. The effects of FUC and GOS on Lactobacillus casei DM8121 were analyzed in vitro. RESULTS: In rats, GOS and FUC supplementation significantly improved serum total cholesterol, low-density lipoprotein cholesterol, lipopolysaccharide, serum total bile acid, hepatic tissue steatosis, aortic arch injury, gut microbiota, serum high-density lipoprotein cholesterol, cholesterol 7-alpha hydroxylase expression in the liver, and BSH activity in the small intestinal contents. In an in vitro experiment, GOS and FUC supplementation significantly increased L. casei DM8121's BSH activity. CONCLUSIONS: In rats, FUC and GOS supplementation improved serum dyslipidemia, gut microbiota, BSH activity, and bile acid metabolism-related pathways. In vitro, GOS and FUC supplementation increased L. casei DM8121's BSH activity.

Antitumor effects of ß-elemene via targeting the phosphorylation of insulin receptor.

Ewing sarcoma family tumors (ESFTs) are a group of aggressive and highly metastatic tumors lacking efficient therapies. Insulin-like growth factor 1 receptor (IGF1R) blockade is one of the most efficient targeting therapy for ESFTs. However, the appliance is obstructed by drug resistance and disease recurrence due to the activation of insulin receptor (IR) signaling induced by IGF1R blockade. Herein ß-elemene, a compound derived from natural plants, exhibited a remarkable proliferation repression on ESFT cells, which was weakened by a caspase inhibitor Z-VAD. ß-elemene in combination with IGF1R inhibitors enhanced markedly the repression on cellular proliferation and mTOR activation by IGF1R inhibitors and suppressed the PI3K phosphorylation induced by IGF1R inhibitors. To investigate the mechanisms, we focused on the effects of ß-elemene on IR signaling pathway. ß-elemene significantly suppressed the insulin-driven cell growth and the activation of mTOR and PI3K in tumor cells, while the toxicity to normal hepatocytes was much lower. Further, the phosphorylation of IR was found to be suppressed notably by ß-elemene specifically in tumor cells other than normal hepatocytes. In addition, ß-elemene inhibited the growth of ESFT xenografts in vivo, and the phosphorylation of IR and S6 ribosomal protein was significantly repressed in the ß-elemene-treated xenografts. These data suggest that ß-elemene targets IR phosphorylation to inhibit the proliferation of tumor cells specifically and enhance the effects of IGF1R inhibitors. Thus, this study provides evidence for novel approaches by ß-elemene alone or in combination with IGF1R blockades in ESFTs and IR signaling hyperactivated tumors.

Herbal medicine Gan­fu­kang downregulates Wnt/Ca2+ signaling to attenuate liver fibrogenesis in vitro and in vivo.

The present study was designed to verify the effect of the Chinese prescription Gan­fu­kang (GFK) on the treatment of liver fibrosis, and to investigate its underlying mechanisms. Liver fibrosis was established in rats by the subcutaneous administration of 0.5 mg/kg carbon tetrachloride (CCl4) twice a week for 8 weeks. Subsequently, the rats were divided into four CCl4 groups, which were treated daily with vehicle and GFK (31.25, 312.5 and 3,125 mg/kg/day) orally between weeks 9 and 20. The inhibitory action of GFK­medicated serum on platelet­derived growth factor (PDGF)­stimulated HSC­T6 cells was also investigated. Biochemical parameters, hydroxyproline (Hyp) content and histological changes to the liver were measured. Reverse transcription­quantitative polymerase chain reaction, western blotting and immunohistochemistry were used to examine the expression of α­smooth muscle actin (α­SMA), PDGF­BB, PDGF receptor ß, collagen type I and II, and the Wnt/Ca2+ signaling pathway. The results showed that GFK significantly alleviated the histological changes, decreased the content of Hyp in the liver and improved liver function in rats. In addition, GFK and GFK­medicated serum effectively inhibited collagen deposition, reduced the expression of α­SMA and downregulated the Wnt/Ca2+ signaling pathway in vivo and in vitro, respectively, as well as cell viability (P<0.05). These results indicated that GFK was effective in attenuating liver injury and fibrosis through downregulation of the Wnt/Ca2+ signaling pathway.

Biological properties of 6-gingerol: a brief review.

Numerous studies have revealed that regular consumption of certain fruits and vegetables can reduce the risk of many diseases. The rhizome of Zingiber officinale (ginger) is consumed worldwide as a spice and herbal medicine. It contains pungent phenolic substances collectively known as gingerols. 6-Gingerol is the major pharmacologically-active component of ginger. It is known to exhibit a variety of biological activities including anticancer, anti-inflammation, and anti-oxidation. 6-Gingerol has been found to possess anticancer activities via its effect on a variety of biological pathways involved in apoptosis, cell cycle regulation, cytotoxic activity, and inhibition of angiogenesis. Thus, due to its efficacy and regulation of multiple targets, as well as its safety for human use, 6-gingerol has received considerable interest as a potential therapeutic agent for the prevention and/or treatment of various diseases. Taken together, this review summarizes the various in vitro and in vivo pharmacological aspects of 6-gingerol and the underlying mechanisms.

Effects of Ganfukang on expression of connective tissue growth factor and focal adhesion kinase/protein kinase B signal pathway in hepatic fibrosis rats.

OBJECTIVE: To investigate the effect of Ganfukang (GFK) on connective tissue growth factor (CTGF) and focal adhesion kinase (FAK)/protein kinase B (PKB or Akt) signal pathway in a hepatic fibrosis rat model and to explore the underlying therapeutic molecular mechanisms of GFK. METHODS: Fifty SD rats were randomly divided into five groups as follows: the control group, the model group (repeated subcutaneous injection of CCl4), and the three GFK treatment groups (31.25, 312.5, and 3125 mg/kg, intragastric administration). Reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemistry were used to examine the expression of CTGF, integrin α5, integrin ß1, FAK/Akt signal pathway, cyclinD1, and collagen in the different-treated rats. RESULTS: GFK attenuated the up-regulation of CTGF, integrin α5, and integrin ß1 in hepatic fibrosis rats and suppressed both the phosphorylation of FAK and the phosphorylation of Akt simultaneously (P<0.01). At the same time, the expression of cyclinD1, collagen I, and collagen III was decreased by GFK significantly (P<0.01). CONCLUSIONS: CTGF and FAK/Akt signal pathway were activated in the CCl4-induced hepatic fibrosis rats, which contribute to increased expression of cyclinD1 and collagen genes. The mechanisms of the anti-fibrosis activity of GFK may be due to its effects against CTGF and FAk/Akt signal pathway.

Protective effect of the herbal medicine Gan­fu­kang against carbon tetrachloride­induced liver fibrosis in rats.

The aim of the present study was to investigate the effect of a herbal medicine formula, Gan-fu-kang (GFK), on the treatment of liver fibrosis in rats and the mechanisms via which it exerts its effect. Liver fibrosis was induced in rats by subcutaneous injection of carbon tetrachloride (CCl4) at 0.5 mg/kg body weight, twice a week for 8 weeks. The rats were randomly selected to receive saline or GFK at 31.25, 312.5 or 3,125 mg/kg body weight/day between weeks 9 and 20. An additional group of rats without CCl4 injection was used as the baseline. In the liver fibrosis model rats, an increase in plasma liver enzymes, fibrotic markers in serum and liver fibrosis, production of α-smooth muscle actin, matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-1, synthesis of collagen and activation of the Wnt/ß-catenin signaling pathway were observed. GFK administration was found to significantly reduce these changes. Results of this study demonstrate that GFK has a protective and therapeutic effect on liver fibrosis induced by CCl4, which may be associated with its inhibitory activity on HSC proliferation and collagen synthesis, effectively downregulating Wnt/ß-catenin signaling.

[Influence of Lipoplus fat emulsion on postoperative nutritional status and early inflammatory response in patients with gastrointestinal malignancies].

OBJECTIVE: To investigate the effect of Lipoplus on postoperative nutritional status and inflammatory response in patients with gastrointestinal malignancies. METHODS: Sixty-four patients with gastrointestinal malignancies were randomized using random digit table to receive isonitrogenous, isocaloric total parenteral nutrition for 5 days including either Lipoplus or Lipofundin with 32 patients in each group. Blood samples were obtained before the surgery, postoperative days 1, 2, 3, and 6 to evaluate the nutritional status(prealbumin, retinol binding protein, and nitrogen balance) and inflammatory response [C-reaction protein(CRP), and leukotriene(LTB) 5, LTB4]. The incidence of postoperative systemic inflammatory response syndrome(SIRS), infection, postoperative complications, mortality, APACHEII score, length of hospital stay and other clinical indicators were recorded. RESULTS: On postoperative day 1, prealbumin and retinol binding protein were significantly lower as compared to preoperative levels. These parameters increased significantly(P<0.05) on postoperative day 6 and the nitrogen balance was positive. On postoperative day 6, CRP was significantly lower in both groups as compared to postoperative day 3 (P<0.05), and the decrease was more prominent in Lipoplus than Lipofundin(P<0.05). There was a significant increase in LTB5/LTB4 as compared to postoperative day 1(P<0.05) in the Lipoplus group, however the increase was not statistically significant in the Lipofundin group(P>0.05). The incidence of postoperative infection was significantly lower in the Lipoplus group(3.1% vs. 6.3%, P<0.05), as was that of SIRS(9.4% vs. 15.6%, P<0.05). The APACHEII score was higher in the Lipoplus group but the difference was not statistically significant(3.6±2.0 vs. 3.3±2.1, P>0.05). The length of hospital stay was significantly shorter in Lipoplus group[(6.4±1.1) d vs. (8.2±1.3) d, P<0.05]. CONCLUSION: Lipoplus can improve the postoperative nutritional status and minimize the inflammatory response in patients with gastrointestinal malignancies.

[Determination of p-coumaric acid in Hedyotis diffusa Willd. from different sources by reversed-phase high performance liquid chromatography].

A method for the determination of p-coumaric acid in Hedyotis diffusa Willd. from different sources by reversed-phase high performance liquid chromatography (RP-HPLC) has been developed. p-Coumaric acid was successfully separated on a Diamonsil ODS column (4.6 mm i.d. x 250 mm, 5 microm) at ambient temperature, using a mixture of CH3CN-20 mmol/L NH4Ac (pH 4.0) (15:85, v/v) as mobile phase and detection at 308 nm. The flow rate was 1.0 mL/min. There was good linear relationship (r = 0.9996) between the mass concentration and the peak area of p-coumaric acid in the range of 4.04 - 202 mg/L. The recoveries were found to be in the range of 97.4% - 102.2%. The results of the experiments have demonstrated that the established method is rapid and simple with good accuracy and reproducibility. The method is suitable for use in quality control of Hedyotis diffusa Willd. from different sources.

[Determination of the content of geniposide in xinxue granules by high performance liquid chromatography].

A high performance liquid chromatographic method has been developed for the determination of geniposide in Xinxue granules. Geniposide was extracted by ultrasonic extraction for 30 min. The chromatographic separation was carried out on a Diamonsil C18 column (200 mm x 4. 6 mm i.d., 5 microm) with a mobile phase consisting of an acetonitrile-water (15:85, v/v) mixture. The detection wavelength was set at 238 nm. The calibration curve was linear in the ranged of 25 - 400 mg/L for geniposide. The average recovery of geniposide was 101.2% with a relative standard deviation (RSD) of 1.6% and contents of geniposide in the granules ranged from 0.841 to 0.923 mg/g. This method is simple, reliable and suitable for quality control of Xinxue granules.