Green Synthesis of Gold Nanoparticles from Polygahatous Polysaccharides and Their Anticancer Effect on Hepatic Carcinoma through Immunoregulation.

Hepatic carcinoma is one of the leading causes of morbidity and mortality among all cancers, but no effective treatment measures have been developed. Herein, polystyrene polysaccharide (PSP) extracted from Polygonatum was used to synthesize gold nanoparticles (PSP-AuNPs) by heating and reduction methods, and the characteristics of the PSP-AuNPs were detected after successful synthesis. In vitro, the immunoregulatory effects of PSP-AuNPs were studied by testing the concentrations of NO, TNF-α, and IL-12p70 in the culture media of PSP-AuNPs-treated RAW264.7 macrophages, and the effect of biocompatibility on the viability of RAW264.7 macrophages and L02 cells was studied via a CCK-8 assay. In vivo, tumor-bearing mice were established and treated with PSP-AuNPs, and the anticancer effects were studied by detecting trends in tumor volume, tumor inhibition rate, and tumor cell proliferation index. Immunoregulation was assessed by evaluating the serum levels of TNF-α and IL-10, the CD4+/CD8+ lymphocyte ratio in peripheral blood and the spleen and thymus indices; toxicity was investigated by measuring body weight, liver and renal function indices. The results showed that PSP-AuNPs could regulate immune function both in vitro and in vivo with almost no toxicity. PSP-AuNPs exhibited excellent anticancer effects on hepatic carcinoma in vivo. The anticancer effect could be strengthened, and the toxicity could be reduced by the combined use of PSP-AuNPs and ADM. In conclusion, PSP-AuNPs could be effective as a therapy and adjuvant therapy for treating hepatic carcinoma, providing potential treatment strategies for this disease.

Ferritinophagy-mediated ferroptosis facilitates methotrexate-induced hepatotoxicity by high-mobility group box 1 (HMGB1).

BACKGROUND AND AIM: Hepatotoxicity is a well-defined reaction to methotrexate (MTX), a drug commonly used for the treatment of rheumatoid arthritis and various tumours. We sought to elucidate the mechanism underlying MTX-induced hepatotoxicity and establish a potentially effective intervention strategy. METHODS: We administered MTX to liver cells and mice and assessed hepatotoxicity by cell viability assay and hepatic pathological changes. We determined ferroptosis and ferritinophagy by detecting ferroptosis-related markers and autophagic degradation of ferritin heavy chain 1 (FTH1). RESULTS: We have shown that hepatocytes treated with MTX undergo ferroptosis, and this process can be attenuated by ferroptosis inhibitors. Interestingly, NCOA4-mediated ferritinophagy was found to be involved in MTX-induced ferroptosis, which was demonstrated by the relief of ferroptosis through the inhibition of autophagy or knockdown of Ncoa4. Furthermore, MTX treatment resulted in the elevation of high-mobility group box 1 (HMGB1) expression. The depletion of Hmgb1 in hepatocytes considerably alleviated MTX-induced hepatotoxicity by limiting autophagy and the subsequent autophagy-dependent ferroptosis. It is noteworthy that glycyrrhizic acid (GA), a precise inhibitor of HMGB1, effectively suppressed autophagy, ferroptosis and hepatotoxicity caused by MTX. CONCLUSION: Our study shows the significant roles of autophagy-dependent ferroptosis and HMGB1 in MTX-induced hepatotoxicity. It emphasizes that the inhibition of ferritinophagy and HMGB1 may have potential as a therapeutic approach for preventing and treating MTX-induced liver injury.

Promoting GSDME expression through DNA demethylation to increase chemosensitivity of breast cancer MCF-7 / Taxol cells.

OBJECTIVE: Breast cancer is the most common and high-incidence cancer in women. It is mainly treated by surgery combined with chemoradiation. The main challenge in treating breast cancer patients is developing resistance to chemotherapeutics, so it is urgent to find potential strategies that can improve the chemotherapy effect of patients. In this study, we aimed to explore the role of GSDME methylation in the sensitivity of chemotherapy for breast cancer. METHODS: Here, we identified breast cancer MCF-7 / Taxol cells models using quantitative real-time PCR (qRT-PCR), Western blotting (WB), and cell counting kit-8 (CCK-8) analyses. Epigenetic changes in it were detected by Methylated DNA immunoprecipitation-sequencing and methylation-specific PCR. The expression level of GSDME in breast cancer cells was observed by qPCR and WB analyses. CCK-8 and colony formation assay were used to detect cell proliferation. Finally, pyroptosis was detected by LDH assay, flow cytometry, and WB analyses. RESULTS: Our results indicate that ABCB1 mRNA and p-GP expression are significantly increased in breast cancer MCF-7 / Taxol cells. GSDME enhancer methylation was found in drug-resistant cells and was associated with the down-regulation of GSDME expression. After treatment with decitabine (5-Aza-2'-deoxycytidine), the demethylation of GSDME induced the occurrence of pyroptosis and thereby inhibited the proliferation of MCF-7 / Taxol cells. We found that the upregulation of GSDME enhances the chemosensitivity of MCF-7 / Taxol cells to paclitaxel by inducing pyroptosis. CONCLUSION: Taken together, we identified decitabine increases GSDME expression through DNA demethylation and induces pyroptosis, thus increasing the chemosensitivity of MCF-7 / Taxol cells to Taxol. Use of decitabine / GSDME / pyroptosis-based treatment strategies may be a new way to overcome the resistance of breast cancer to paclitaxel chemotherapy.

The molecular characteristics, diagnosis, and treatment of macrolide-resistant Mycoplasma pneumoniae in children.

The purpose of this study is to review the molecular characteristics, the diagnosis, and treatment of the widespread infection of macrolide-resistant Mycoplasma pneumoniae (M. pneumoniae; MRMP) in children, thus providing a better knowledge of this infection and presenting the associated problems. Single point mutations in the V region of the 23S rRNA gene of M. pneumoniae genome are associated with macrolide resistance. P1-1, MLVA4-5-7-2, and ST3 are usually the predominated genetic types in the M. pneumoniae epidemics. The short-term two times serological IgM (or together with IgG) test in the acute stage can be used for confirmation. Combined serological testing and PCR might be a more prudent method to reduce macrolide consumption and antibiotic selective pressure in a clinical setting. Molecular methods for the detection of single-nucleotide mutations in the V region of the 23S rRNA gene can be used for the diagnosis of MRMP. The routine use of macrolide for the treatment of macrolide-sensitive Mycoplasma pneumoniae (MSMP) infections can get good effect, but the effects are limited for severe MRMP infections. Additional corticosteroids may be required for the treatment of severe MRMP infections in children in China during the era of MRMP.

Altered DNA methylation and transcription of WNT2 and DKK1 genes in placentas associated with early-onset preeclampsia.

PURPOSE: The purpose of this study was to characterize the changes in DNA methylation and transcription of WNT2 and DKK1 genes in placentas associated with early-onset preeclampsia. METHODS: The study includes three groups: patients with early-onset preeclampsia, normotensive preterm and term births. Placental tissues were collected and pyrosequencing was performed on DKK1 and WNT2 proximal promoters. Transcriptional levels of DKK1 and WNT2 genes were determined with real-time PCR. RESULTS: DKK1 gene methylation levels were lower in placentas associated with early-onset preeclampsia compared to those associated with preterm birth (P<0.05). DKK1 mRNA expression was higher in early-onset preeclampsia placentas than those in preterm placentas (P < 0.05). WNT2 mRNA expression in early-onset preeclamptic placentas was lower than that in other two groups (P < 0.05). In the preterm and early-onset preeclampsia groups, the mRNA levels for WNT2 and DKK1 were negatively correlated (P < 0.05). In all the subjects, the levels of DKK1 mRNA and methyaltion were negatively correlated (P < 0.05). CONCLUSION: Decreased methylation of DKK1 promoter in early-onset preeclamptic placenta tissues may up-regulate the expression of DKK1. The increased expression of DKK1 and decreased expression of WNT2 may be involved in the pathogenesis of early-onset preeclampsia.

HMGB1-RAGE signaling pathway in pPROM.

OBJECTIVE: Increased inflammation of the placenta is considered as a risk factor and a promoter of preterm premature rupture of the membranes (pPROM). High-mobility group box 1 (HMGB1) is a recently identified inflammatory cytokine, and HMGB1-RAGE signaling pathway has been associated with many pathophysiological processes. This study aims to reveal the mechanisms of HMGB1-RAGE signaling pathway in pPROM. MATERIALS AND METHODS: The mRNA levels of relative gene of HMGB1 pathway, HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2, were analyzed by real-time PCR in placentas collected from 60 normal term women, 60 women with PROM and 60 women with pPROM. Additionally, levels of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 protein were detected in frozen placental specimens by western blot, and the locations of HMGB1, RAGE and NF-κBp65 were evaluated in the well-characterized tissue microarray (TMA) by immunohistochemistry. ELISA was further used to detect HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 level in maternal and cord serum. RESULTS: Compared with normal term and PROM women, we found that (1) The mRNA expressions of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in HMGB1-RAGE pathway of pPROM placentas were higher. (2) The protein levels of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in pPROM placentas were higher. (3) HMGB1 and RAGE immunoreactivity in pPROM placenta TMA were increased in the cytoplasm of syncytiotrophoblast (STB), extravillous trophoblast (EVT) and mesenchymal cells, while NF-κBp65 was enhanced in the nucleus of STB and EVT. (4) Maternal serum concentrations of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 in pPROM group were greater. (5) Cord serum concentrations of HMGB1, RAGE, NF-κBp65, MMP-9 and MMP-2 among the 3 groups had no significant differences. CONCLUSION: HMGB1 nuclear-cytoplasmic translocation in pPROM placenta may lead to the binding of HMGB1 to its receptor RAGE, resulting in provoking NF-κBp65 activity, and then inducing the release of MMP-9 and MMP-2, which all above activities contributed to the process of pPROM. Consequently, HMGB1-RAGE signaling pathway may be involved in the pathogenesis of pPROM.