Artesunate restores mitochondrial fusion-fission dynamics and alleviates neuronal injury in Alzheimer's disease models.

Alzheimer's disease (AD) remains a leading cause of dementia and no therapy that reverses underlying neurodegeneration is available. Recent studies suggest the protective role of artemisinin, an antimalarial drug, in neurological disorders. In this study, we investigated the therapeutic potential of artesunate, a water-soluble derivative of artemisinin, on amyloid-beta (Aß)-treated challenged microglial BV-2, neuronal N2a cells, and the amyloid precursor protein/presenilin (APP/PS1) mice model. We found that Aß significantly induced multiple AD-related phenotypes, including increased expression/production of pro-inflammatory cytokines from microglial cells, enhanced cellular and mitochondrial production of reactive oxygen species, promoted mitochondrial fission, inhibited mitochondrial fusion, suppressed mitophagy or biogenesis in both cell types, stimulated apoptosis of neuronal cells, and microglia-induced killing of neurons. All these in vitro phenotypes were attenuated by artesunate. In addition, the over-expression of the mitochondrial fission protein Drp-1, or down-regulation of the mitochondrial fusion protein OPA-1 both reduced the therapeutic benefits of artesunate. Artesunate also alleviated AD phenotypes in APP/PS1 mice, reducing Aß deposition, and reversing deficits in memory and learning. Artesunate protects neuronal and microglial cells from AD pathology, both in vitro and in vivo. Maintaining mitochondrial dynamics and simultaneously targeting multiple AD pathogenic mechanisms are associated with the protective effects of artesunate. Consequently, artesunate may become a promising therapeutic for AD.

Bilobalide alleviates neuroinflammation and promotes autophagy in Alzheimer's disease by upregulating lincRNA-p21.

EGb 761 has some protective effects on AD and can improve the cognitive functions of AD mice. However, the underlying molecular mechanisms are unknown. Here, we investigated the function of bilobalide, the effective component of EGb 761, in neuroinflammation and autophagy during AD. LPS-treated BV-2 cells were used as an in vitro model for neuroinflammation. The APP/PS1 AD mouse line was used to examine the function of bilobalide in AD. ELISA and qRT-PCR were used to measure the levels of proinflammatory cytokines, including TNF-α, IL-6 and IL-1ß. Western blotting was employed to determine the protein levels of p-p65, iNOS, COX-2, LC3, beclin-1, p62 and p-STAT3. Immunostaining was applied to examine the number of autophagosomes. LPS treatment induced inflammatory responses and inhibited autophagy in BV-2 cells. Bilobalide suppressed LPS-induced neuroinflammation and promoted autophagy. Furthermore, bilobalide treatment increased the lincRNA-p21 levels, which suppressed STAT3 signalling. Knockdown of lincRNA-p21 reversed the effects of bilobalide. Overexpression of lincRNA-p21 promoted autophagy and inhibited neuroinflammation as well while STAT3 inhibitor blocked the effects of si-lincRNA-p21. In vivo experiments revealed that bilobalide improved the learning and memory capabilities of APP/PS1 AD mice. Bilobalide improves the cognitive functions of APP/PS1 AD mice. Mechanistically, bilobalide suppresses inflammatory responses and promotes autophagy possibly by upregulating lincRNA-p21 levels.

Two novel chiral tetranucleate copper-based complexes: crystal structures, nanoparticles, and inhibiting angiogenesis and the growth of human breast cancer by regulating the VEGF/VEGFR2 signal pathway in vitro.

The single crystals of two novel copper(ii)-based complexes containing l-methioninol-derived Schiff bases were obtained and characterized. The nanoparticles of these complexes were prepared and their cellular uptake was measured in MDA-MB-231 cells and HUVECs. It was found that these complexes could remarkably induce apoptosis, inhibit proliferation, suppress migration and metastasis, and inhibit angiogenesis and the growth of triple-negative breast cancer derived from MDA-MB-231 cells in vitro. Meanwhile, these complexes exhibit anticancer and antiangiogenic functions by activating the important protein molecules VEGFR2, FAK, AKT and Erk1/2 or their phosphorylated molecules p-VEGFR2, p-FAK, p-AKT, and p-Erk1/2 in the VEGF/VEGFR2 signaling pathway, collapsing the mitochondrial membrane potential, and damaging the level of reactive oxygen species.

Anti-cancer activities of metal-based complexes by regulating the VEGF/VEGFR2 signaling pathway and apoptosis-related factors Bcl-2, Bax, and caspase-9 to inhibit angiogenesis and induce apoptosis.

Three novel single crystals of the metal-based complexes Cu-1, Cu-2, and Co-1 were obtained and characterized. Compared with Cu-2 and Co-1, Cu-1 showed remarkable activities of anti-cervical cancer, anti-cisplatin-resistant non-small cell lung cancer and anti-angiogenesis by downregulating the expressions of important proteins in the VEGF/VEGFR2 signaling pathway to inhibit angiogenesis and cancer cell proliferation, induce apoptosis, and suppress migration and metastasis. Moreover, Cu-1 dramatically inhibited the expression of the anti-apoptotic protein Bcl-2 and up-regulated the expressions of the proapoptotic proteins caspase-9 and Bax to induce the apoptosis of tumor cells, simultaneously decreasing the density of endothelial cells to inhibit tumor angiogenesis in cisplatin-resistant tumors.

Prognostic significance of ACP5 expression in patients with lung adenocarcinoma.

INTRODUCTION: Tartrate-resistant acid phosphatase 5 (ACP5), which is essential for bone resorption and osteoclast differentiation, promotes cell motility through the modulation of focal adhesion kinase phosphorylation. This study seeks to elucidate the association of ACP5 expression and the clinicopathologic characteristics of patients with lung adenocarcinoma (AD). METHODS: The expression of ACP5 was measured by Immunohistochemistry and Western blot analysis in lung AD and matched tumor-adjacent tissues, and the χ2 test was applied to analyze the correlation between ACP5 expression and clinicopathologic features. Using the Kaplan-Meier method, univariate and multivariate regression analysis was to explore the correlation between ACP5 expression and overall survival (OS). RESULTS: We found that ACP5 was frequently upregulated in lung AD tissues. The high expression of ACP5 was significantly related to lymph node status, tumor-node-metastasis (TNM) stage, and differentiation. From the results of univariate survival analysis, it indicated that the patients with high expression of ACP5 expression had a significantly lower OS than the patients with low expression of ACP5 expression. As it showed in Multivariate Cox regression analysis, the high expression of ACP5 expression was an independent prognostic factor for OS. CONCLUSIONS: Our results suggest that high expression of ACP5 correlates with tumor progression and may serve as a potential prognostic biomarker in lung AD.

Association between MDR1 gene of gastrointestinal tumors, the expression of P-glycoprotein and resistance to chemotherapeutic drugs.

The aim of the present study was to examine and discuss the association between multidrug resistance 1 gene (MDR1) of gastrointestinal tumors, the expression of P-glycoprotein and resistance to chemotherapeutic drugs. In this study, 126 cases of patients with gastrointestinal tumors admitted to hospital from February 2013 to February 2015 were selected. The expression levels of MDR1 gene were obsreved in the control population and patients before and after treatment by fluoresecent quantitative PCR. The protein expression level of P-glycoprotein was determined using western blotting and enzyme-linked immunosorbent assay. In addition, drug resistance was assessed by ATP-TCA chemosensitivity experiments. The results showed that before treatment, the expression of mRNA in MDR1 of tissues of gastrointestinal tract of the 126 cases was 108-fold larger than that of the gastrointestinal tract of the controls (p<0.05), P-glycoprotein was 87-fold larger than the expression level of the controls (p<0.05). The sensitivity of 126 tumor tissues to different chemotherapeutic drugs was determined, and the results showed that most of the tumor tissues were sensitive to chemotherapeutic drugs, and the sensitivity rate reached 96.4%. Following chemotherapy, the expression of mRNA in MDR1 of tumor tissues and the expression of P-glycoprotein decreased (p<0.05). In conclusion, the MDR1 gene and P-glycoprotein have a positive correlation with the occurrence of gastrointestinal tumors, and a negative correlation between the MDR1 gene and P-glycoprotein with resistance of chemotherapeutic drugs. Therefore, the MDR1 gene and P-glycoprotein can be used as references in the identification and diagnosis of gastrointestinal tumors.

Combination of temsirolimus and adriamycin exhibits an enhanced antitumor effect in hepatocellular carcinoma.

OBJECTIVE: The oncogenic PI3K/Akt/mTOR pathway is frequently activated in hepatocellular carcinoma (HCC). The aim of this study is to investigate the anti-HCC effect of combination of temsirolimus, an mTOR inhibitor, and adriamycin, a routinely used drug for treating HCC. METHODS AND MATERIALS: Proliferation of HCC cells exposure to temsirolimus, adriamycin, and their combination was determined using MTT assay in vitro as well as in a nude mice model in vivo. Cell apoptosis was examined using flow cytometry. Expressions of apoptosis-related proteins including caspase-9, -3, PARP, Bax, and Bcl-2 were determined using Western blotting. RESULTS: Temsirolimus plus adriamycin showed an enhanced inhibitory effect on cell proliferation compared to temsirolimus or adriamycin in HCC cells PLC/PRF/5, BEL7402, and HuH7 in vitro. The drug combination solicited a higher percentage of apoptosis cells and induced higher levels of cleaved caspase-9, -3, and PARP than temsirolimus or adriamycin used alone. The ratio of Bax/Bcl-2 was increased in cells exposed to the combination treatment. The enhanced anti-tumor effect of this drug combination was verified in a nude mice model. We also observed that half doses of temsirolimus and adriamycin used in combination achieved a comparable tumor growth inhibitor rate with full dose of temsirolimus or adriamycin used alone. CONCLUSION: Temsirolimus plus adriamycin exhibited an enhanced antitumor effect in HCC and this drug combination might have a potential value in treatment of HCC. Studies are warranted to comprehensively evaluate the efficacy and safety of this regimen in the future.

[Efficacy observation on osteoarthritis of the knee treated with the ultrastructural acupotomy therapy at the counter-Ashi points].

OBJECTIVE: To compare the difference in the efficacy on osteoarthritis of the knee between the ultrastructural acupotomy therapy at the counter-Ashi points and the conventional acupuncture-moxibustion therapy. METHODS: Sixty cases were randomly devided into an ultrastructural acupotomy therapy group (group A) and a conventional acupuncture-moxibustion group (group B), 30 cases in each one. In the group A, the ultrastructural acupotomy therapy was applied to the counter-Ashi points in which pain was alleviated or disappeared on pressure. The treatment was given once a week, lasting for 1 month. In the group B, the acupuncture-moxibustion therapy was applied to Dubi (ST 35), Neixiyan (EX-LE 4), Zusanli (ST 36), Yanglingquan (GB 34), etc. The treatment was given once daily, lasting for 1 month. Before and after treatment, the Visual Analogue Scale (VAS), Lysholm knee joint scale and the affected knee joint flexion angle were observed in the two groups. The clinical efficacy was compared between the two groups. RESULTS: After treatment, the scores of VAS, Lysholm knee joint scale and the affected knee joint flexion angle were improved obviously as compared with those before treatment in either group (P < 0.01, P < 0.05). The results of them in the group A were superior apparently to those in the group B (all P < 0.05). The total effective rate in the group A was superior to that in the group B [80.0% (24/30) vs 60.0% (18/30), P < 0.05]. CONCLUSION: The ultrastructural acupotomy therapy at the counter-Ashi points achieves the superior clinical efficacy on osteoarthritis of the knee as compared with the conventional acupuncture therapy. It relieves pain and improves the motion range of knee joint effectively. It is simple in operation and less in treatment frequency.

[Effect of acupuncture at "Zusanli" (ST 36 and "Taichong" (LR 3) on gastrointestinal hormone levels in rats with diarrhea type irritable bowel syndrome].

OBJECTIVE: To observe the effect of acupuncture of "Zusanli" (ST 36) and "Taichong" (LR 3) on gastrointestinal hormone levels in diarrhea type irritable bowel syndrome (IBS-D) rats so as to provide experimental evidence for acupuncture treatment of IBS-D. METHODS: Forty-eight Wistar rats were randomly divided into control, model, acupuncture and medication groups, with 12 rats in each group. IBS-D model was established by chronic mild restraining stress combined with isolated raising and intragastric administration of Folium Sennae (10 mL/kg). For rats of the acupuncture group, bilateral "Zusanli" (ST 36) and "Taichong" (LR 3) were punctured and stimulated for 30 min, once daily for 14 days. The rats of the medication group were treated by gavage of Pinaverin bromide, once daily for 14 days, and those of the control and model groups were given with equal volume of saline. Somatostatin (SS), substance P (SP) and vasoactive intestinal peptide (VIP) in the rats' plasma and ileum tissues were detected by sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared with the control group, the body weight and food intake quantity were reduced significantly, while the diarrhea index was increased significantly in the model group (P < 0.05, P < 0.01). In comparison with the model group, the body weight and food intake volume in both acupuncture and medication groups were increased remarkably after the treatment (P < 0.05, P < 0.01) whereas the diarrhea index in the latter two groups was reduced obviously (P < 0.01). Compared with the control group, plasma VIP level was increased significantly in the model group (P < 0.01), while those in both acupuncture and medication groups were decreased remarkably compared with the model group (P < 0.05). No significant changes of plasma SP and SS levels were found after modeling and after both acupuncture and medication treatments. In comparison with the control group, ileum SP, VIP and SS levels in the model group were up-regulated significantly (P < 0.01), while those in both acupuncture and medication groups were down-regulated considerably compared with the model group (P < 0.01). No significant differences were found between acupuncture and medication groups in the levels of ileum SP, VIP and SS (P > 0.05). CONCLUSION: Acupuncture of ST 36 and LR 3 can effectively down-regulate ileum SP, VIP and SS levels in IBS-D rats, which may contribute to its effect in relieving IBS-D.