M1 macrophages-derived exosomes miR-34c-5p regulates interstitial cells of Cajal through targeting SCF.

Slow transit constipation (STC) is a gastrointestinal disorder characterized by abnormal prolonged colonic transit time, which affects the life quality of many people. The decrease number of interstitial cells of Cajal (ICCs) is involved in the pathogenesis of STC. However, the molecular mechanism of loss of ICCs in STC remains unclear, making it difficult to develop new agents for the disease. In this study, we investigated the mechanism of decreasing ICCs in the pathogenesis of STC. We constructed the STC model rats by using atropine and diphenoxylate. A series of methods were used including immunofluorescence and immunochemistry staining, western blot, qRT-PCR, exosomes extraction and exosomes labeling. The results indicate that ICCs decreased in the STC rats accompanied with the macrophages activation. Further studies suggested that macrophages decreased the cell viability of ICCs by secretion exosomes containing miR-34c-5p. miR-34c5p targeted the 3Ꞌ -UTR of stem cell factor(SCF) mRNA and regulated the expression of SCF negatively. In conclusion, we demonstrated a novel regulatory mechanism of ICCs cell viability in STC. We found that exosome miR-34c-5p mediate macrophage-ICCs cross-talk. M1 macrophages derived exosomes miR-34c-5p decreased ICCs cell viability by directly targeting SCF.

Prognostic value of CD133 expression in cancer patients treated with chemoradiotherapy: a meta-analysis.

Many studies evaluated the correlations of CD133 expression with the clinical outcomes in patients treated with chemoradiotherapy (CRT) but yielded controversial results. This meta-analysis was performed to identify the impacts of CD133 expression on the prognosis of cancer patients treated with CRT. Electronic databases updated up to March 2014 were searched to find relevant studies. Relevant literatures without any language restrictions were searched via electronic databases as follows: Web of Science (1945 ~ 2013), the Cochrane Library Database (Issue 12, 2013), PubMed (1966 ~ 2013), EMBASE (1980 ~ 2013), CINAHL (1982 ~ 2013), and the Chinese Biomedical Database (CBM) (1982 ~ 2013). STATA software was used for the current meta-analysis. Hazard ratios (HR) and its corresponding 95% confidence interval (95% CI) were calculated. Six studies were identified with a total of 470 cancer patients treated with CRT. The meta-analysis results showed that CD133-positive patients had poorer overall survival (OS) than that of CD133-negative patients (HR = 2.13, 95% CI = 1.20 ~ 3.07, P < 0.001). Furthermore, CD133-positive patients displayed shorter disease-free survival (DFS) than that of CD133-negative patients (HR = 1.74, 95% CI = 0.08 ~ 3.40, P = 0.039). Ethnicity-stratified analysis indicated that CD133 expression positively correlated with shorter OS among the Japanese, Chinese, and Spanish populations (all P < 0.05). In conclusion, our findings suggest that CD133 expression may be positively correlated with poorer prognosis in cancer patients treated with CRT.

[Effect of qingchang huash recipe on NF-kappaB/Tolls pathway in ulcerative colitis patients and mechanism study].

OBJECTIVE: To observe the effect of Qingchang Huashi Recipe (QHR) on the activation and expressions of nuclear factor kappaB (NF-kappaB), Toll-like receptors (TLRs), and contents of interleukin-8 (IL-8), thus exploring its possible mechanisms for treating ulcerative colitis (UC). METHODS: The HT-29 cells were induced to inflammation model by tumor necrosis factor-alpha (TNF-alpha) and lipopolysaccharides (LPS). HT-29 cells were divided into 6 groups, i.e., the vehicle control group, the model control group, the sulfasalazine (SASP) group, the high dose QHR group, the middle dose QHR group, the low dose QHR group. Effects on the cell growth were detected by MTT. The chemoattractant of macrophages was observed using Transwell. The expressions of NF-kappaB and TLR4 protein were detected using immune cell fluorescence method. The content of IL-8 was detected by ELISA. RESULTS: The growth of cells were not inhibited in each group. Statistical difference existed in each dose QHR group in inhibiting the chemoattractant of macrophages, reducing activation of NF-kappaB, lowing expressions of TLR4 protein, and decreasing the secretion of IL-8, when compared with the model control group (P < 0.05). No statistical difference existed in inhibiting the chemoattractant of macrophages between the high dose QHR group and the vehicle control group (P > 0.05). But its inhibition on NF-kappaB activation was higher in the high dose QHR group than in the SASP group (P < 0.05). CONCLUSION: QHR could obviously attenuate the inflammatory reaction of HT-29 cells, inhibit the chemoattractant of macrophages, reduce the activation of NF-kappaB, lower expressions of TLR-4, and attenuate the secretion of IL-8, which might be one of its mechanisms for treating UC.

Antitumor activity and toxicity relationship of annonaceous acetogenins.

Annonaceous acetogenins (ACGs) are one of the most interesting classes of natural products appearing in the past two decades. Here, we studied the antitumor activity and toxicity relationship of ACGs including annosquamin B (1), bullatacin (2) and annosquatin B (3) in vivo. A single intraperitoneal (i.p.) injection of 100 µg/kg of annosquamin B, bullatacin and annosquatin B did not cause side effects in normal mice. Bullatacin treatment with five doses of 25 and 50 µg/kg in H22 hepatoma cells bearing mice resulted in about 61% reduction in tumor growth with hematologic parameters increased significantly in normal mice. Annosquamin B and annosquatin B treatments with 10 doses of 25, 50 and 100 µg/kg in the H22 hepatoma cells transplantation tumor model mice resulted in maximum 53.7% and 58.7% reduction in tumor growth, respectively, and did not cause severe side effects in normal mice. This study provided the evidence that adjacent bis-THF ACGs showed higher antitumor activity and toxicity than mono-THF and nonadjacent bis-THF ACGs in vivo. Furthermore, it was found that bullatacin led to liver and kidney toxicity via increasing calcium concentration, ROS production, and Bax expression and Bax/Bcl-2 ratio in rats with repeated treatment with bullatacin for 3 weeks.

[Effects of qingchang huashi recipe on the dendritic cells of the colonic mucosa and the mesenteric lymph nodes in experimental colitis rats].

OBJECTIVE: To observe the effects of qingchang huashi recipe (QHR) on the dendritic cells (DCs) of experimental colitis rats, thus exploring its possible mechanisms for treating ulcerative colitis (UC). METHODS: The UC rat model was induced by TNBS/anhydrous alcohol. Forty male Wistar rats were randomly divided into 4 groups, i.e., the normal group, the model group, the QHR group, and the Mesalazine group, 10 in each group. Since the 2nd day of modeling, corresponding medication was respectively administered to each treatment group by gastrogavage for 10 successive days. The number of DCs in the colonic mucosa was observed using iMmunohistochemical assay. The DCs ratio in the mesenteric lymph nodes, and the expressions of surface molecules MHC-II and CD86 were detected using flow cytometry. RESULTS: Compared with the model group, the number of DCs in the colonic mucosa significantly decreased, the expression of MHC-II in the mesenteric lymph nodes significantly decreased in the QHR group and the Mesalazine group, showing statistical difference (P < 0.01). There was no statistical difference between the two groups (P > 0.05). There was no statistical difference in the DCs ratios and the CD86 expression among the 4 groups (P > 0.05). CONCLUSION: QHR could decrease the infiltration of DCs in the colonic mucosa, and suppress the activation of DCs in the mesenteric lymph nodes, which might be one of its mechanisms for treating UC.