Histone deacetylase inhibitors promote epithelial-mesenchymal transition in Hepatocellular Carcinoma via AMPK-FOXO1-ULK1 signaling axis-mediated autophagy.

Hepatocellular carcinoma (HCC) is the third most frequent cause of cancer-related deaths globally because of high metastasis and recurrence rates. Elucidating the molecular mechanisms of HCC recurrence and metastasis and developing effective targeted therapies are expected to improve patient survival. The promising anti-cancer agents for the treatment of hematological malignancies, histone deacetylase inhibitors (HDIs), have limited effects against epithelial cell-derived cancers, including HCC, the mechanisms involved have not been elucidated. Herein, we studied the molecular mechanisms underlying HDI-induced epithelial-mesenchymal transition (EMT) involving FOXO1-mediated autophagy. Methods: The biological functions of HDIs in combination with autophagy inhibitors were examined both in vitro and in vivo. Cell autophagy was assessed using the generation of mRFP-GFP-LC3-expressing cells and fluorescent LC3 puncta analysis, Western blotting, and electron microscopy. An orthotopic hepatoma model was established in mice for the in vivo experiments. Results: Our study provided novel mechanistic insights into HDI-induced EMT mediated by the autophagy AMPK-FOXO1-ULK1-Snail signaling axis. We demonstrated that autophagy served as a pro-metastasis mechanism in HDI-treated hepatoma cells. HDIs induced autophagy via a FOXO1-dependent pathway, and FOXO1 inhibition promoted HDI-mediated apoptosis in hepatoma cells. Thus, our findings provided novel insights into the molecular mechanisms underlying HDI-induced EMT involving FOXO1-mediated autophagy and demonstrated that a FOXO1 inhibitor exerted a synergistic effect with an HDI to inhibit cell growth and metastasis in vitro and in vivo. Conclusion: We demonstrated that HDIs triggers FOXO1-dependent autophagy, which ultimately promotes EMT, limiting the clinical outcome of HDI-based therapies. Our study suggests that the combination of an HDI and a FOXO1 inhibitor is an effective therapeutic strategy for the treatment of HCC.

Crosstalk between autophagy and epithelial-mesenchymal transition and its application in cancer therapy.

Autophagy is a highly conserved catabolic process that mediates degradation of pernicious or dysfunctional cellular components, such as invasive pathogens, senescent proteins, and organelles. It can promote or suppress tumor development, so it is a "double-edged sword" in tumors that depends on the cell and tissue types and the stages of tumor. The epithelial-mesenchymal transition (EMT) is a complex biological trans-differentiation process that allows epithelial cells to transiently obtain mesenchymal features, including motility and metastatic potential. EMT is considered as an important contributor to the invasion and metastasis of cancers. Thus, clarifying the crosstalk between autophagy and EMT will provide novel targets for cancer therapy. It was reported that EMT-related signal pathways have an impact on autophagy; conversely, autophagy activation can suppress or strengthen EMT by regulating various signaling pathways. On one hand, autophagy activation provides energy and basic nutrients for EMT during metastatic spreading, which assists cells to survive in stressful environmental and intracellular conditions. On the other hand, autophagy, acting as a cancer-suppressive function, is inclined to hinder metastasis by selectively down-regulating critical transcription factors of EMT in the early phases. Therefore, the inhibition of EMT by autophagy inhibitors or activators might be a novel strategy that provides thought and enlightenment for the treatment of cancer. In this article, we discuss in detail the role of autophagy and EMT in the development of cancers, the regulatory mechanisms between autophagy and EMT, the effects of autophagy inhibition or activation on EMT, and the potential applications in anticancer therapy.

The relationship between autophagy and the immune system and its applications for tumor immunotherapy.

Autophagy is a genetically well-controlled cellular process that is tightly controlled by a set of core genes, including the family of autophagy-related genes (ATG). Autophagy is a "double-edged sword" in tumors. It can promote or suppress tumor development, which depends on the cell and tissue types and the stages of tumor. At present, tumor immunotherapy is a promising treatment strategy against tumors. Recent studies have shown that autophagy significantly controls immune responses by modulating the functions of immune cells and the production of cytokines. Conversely, some cytokines and immune cells have a great effect on the function of autophagy. Therapies aiming at autophagy to enhance the immune responses and anti-tumor effects of immunotherapy have become the prospective strategy, with enhanced antigen presentation and higher sensitivity to CTLs. However, the induction of autophagy may also benefit tumor cells escape from immune surveillance and result in intrinsic resistance against anti-tumor immunotherapy. Increasing studies have proven the optimal use of either ATG inducers or inhibitors can restrain tumor growth and progression by enhancing anti-tumor immune responses and overcoming the anti-tumor immune resistance in combination with several immunotherapeutic strategies, indicating that induction or inhibition of autophagy might show us a prospective therapeutic strategy when combined with immunotherapy. In this article, the possible mechanisms of autophagy regulating immune system, and the potential applications of autophagy in tumor immunotherapy will be discussed.

Repeat HIV Testing and Incident Rates among Individuals Attending Voluntary Counseling and Testing Clinics in Wuxi, China: A Retrospective Study.

OBJECTIVE: We aimed to elucidate the rates of repeat HIV testing and incident HIV diagnosis, and baseline CD4+ T cell count among individuals attending HIV voluntary counseling and testing (VCT) clinics in Wuxi, China. METHODS: A repeat HIV testing within 12 months was recorded if individuals had their first test with negative results, during 2013-2014 and retested within 12 months. An incident HIV diagnosis was recorded if individuals had their first test with negative results, during 2013-2015 and had a subsequent positive result at any point by the end of 2015. Data on HIV testing and diagnosis among individuals attending 32 VCT clinics from 2013 to 2015 and HIV diagnosis from other clinical services in Wuxi, China, were retrieved. A multivariate logistic regression model was used to analyze factors associated with repeat HIV testing. Cox regression was used to evaluate factors associated with incident HIV diagnosis. RESULTS: From 2013 to 2014, 11,504 individuals tested HIV negative at their first recorded test, with 655 (5.7%) retesting within 12 months. Higher repeat HIV testing within 12 months was associated with male gender [adjusted odds ratio (aOR) = 1.7, 95% confidence interval (CI): 1.4-2.2], risk behaviors [commercial heterosexual behaviors (aOR = 1.4, CI: 1.1-1.6), male-male sexual behaviors (aOR = 3.7, CI: 2.7-4.9)], injection drug use (aOR = 9.9, CI: 6.5-15.1), and having taken HIV tests previously (aOR = 2.0, CI: 1.6-2.4). From 2013 to 2015, 1,088 individuals tested negative on HIV test at their visit and at ⋝ 2 subsequent tests; of them 30 had incident HIV diagnosis. The overall rate of incident HIV diagnosis among all VCT individuals was 1.6 (95% CI: 1.1-2.1) per 100 person-years. Incident HIV diagnosis was associated with male gender [adjusted hazard ratio (aHR) = 8.5, 95% CI: 1.9-38.1], attending hospital-based VCT clinics (aHR = 7.8, 95% CI: 1.1-58.3), and male-male sexual behavior (aHR = 8.4, 95% CI: 1.5-46.7). Individuals diagnosed at VCT clinics had higher CD4+ T cell count compared with those diagnosed at other clinical services (median 407 vs. 326 copies/mm3, P = 0.003). CONCLUSION: VCT individuals in Wuxi, China, had a low repeat HIV testing rate and high HIV incidence. VCT-clinic-based interventions aimed at increasing repeat HIV testing are needed to detect more cases at an earlier stage, especially among individuals at high risk for HIV infection such as men who have sex with men.

[Bacterial Community Composition of Activated Sludge from Coking Wastewater].

Coking wastewater is a kind of highly toxic and refractory organic wastewater, and aerobic activated sludge, which is dominated by bacteria, determines the efficiency of coking wastewater treatment. However, the bacterial community structure of activated sludge from coking wastewater has rarely been reported. 454 sequencing technology was applied to investigate the structure and biodiversity of the bacterial community. The results of thermal cluster analysis and principal component analysis demonstrated the differences in biodiversities of different activated sludge bacterial communities. The bacterial communities were mainly composed of Proteobacteria, Planctomycetes, Acidobacteria, Candidatus Saccharibacteria, Bacteroidetes, Cyanobacteria, Actinobacteria, Chloroflexi, Firmicutes, Thaumarchaeota, Ignavibacteriae, Verrucomicrobia and Unclassified bacteria. Proteobacteria was the dominant phylum and its abundance was 36.00%-76.98%. The primary genera were Thiobacillus, Thauera, Comamonas, Caldimonas, Steroidobacter, Nitrosomonas, Phycisphaera and Gp4. Most of these genera were related with aromatic hydrocarbon degradation, nitrification and denitrification processes. These results provide a theoretical basis for removal mechanism of pollutants in coking wastewater.

[Biodiversity of Thiocyanate-degrading Bacteria in Activated Sludge from Coking Wastewater].

Thiocyanate (SCN-) is one of the main sources of COD in coking wastewater, and SCN- removal efficiency of the aerobic unit impacts the requirement of discharging standard. Microbial population in the activated sludge plays an important role in SCN- removal of coking wastewater treatment. However, the community structure has rarely been reported. Using SCN- as the sole carbon and energy source, the removal of 100 mg·L-1,300 mg·L-1 and 600 mg·L-1 SCN- by activated sludge was studied and 454 sequencing technology was applied to investigate the biodiversity of SCN--degrading bacteria. The results showed that 100-600 mg·L-1 SCN- could be effectively removed by acclimated activated sludge, the higher SCN- concentration, the higher removal efficiency, but the lower bacterial community diversity indices. The bacterial communities in initial and acclimated sludge samples were mainly composed of Proteobacteria, Bacteroidetes, Chlorobi, Planctomycetes, Acidobacteria, Nitrospira, Firmicutes and Unclassified bacteria. Bacteroidetes and Proteobacteria were dominant phyla in the sludge. Thiobacillus was responsible for SCN- biodegradation in coking wastewater, and its abundance in three sludge samples was 3.07%, 8.63% and 0.27%, respectively. When the concentration of SCN- was less than 300 mg·L-1, Thiobacillus was the main degrading bacteria. While at 600 mg·L-1 SCN-, low-abundance degrading bacteria might have synergistic degradation effect. These results have important significance for revealing SCN- removal mechanism in the coking wastewater treatment.

[Survey of current situation of schistosomiasis health education in Wuxi City].

OBJECTIVE: To understand the current situation of schistosomiasis health education in in Wuxi City where schistosomiasis transmission has been interrupted, so as to provide the evidence for formulating the health education strategies. METHODS: Face to face interviews and a professional designed questionnaire were used to collect the information of the current schistosomiasis health education and investigate the awareness of schistosomiasis knowledge in primary and middle schools and in communities. RESULTS: The total awareness rate of schistosomiasis knowledge was 87.7% among 873 students and the figure was 83.0% among 693 community residents. The students who studied in the schools with more than 1 class hour of schistosomiasis health education, completed schistosomiasis health education material or teaching plan, and implementing health education through multiple ways had higher knowledge awareness rates compared with the schools without (χ2 = 291.408, 709.622, 13.751, all P <0.001). The residents living in the communities with schistosomiasis health education through broadcast/TV or square propaganda had a higher knowledge awareness rate compared with the communities without (χ2= 90.772, 47.436, all P < 0.001). CONCLUSION: The awareness rates of schistosomiasis knowledge among both students and community residents in Wuxi City are low. Therefore, the schistosomiasis control health education should be strengthened.

[Distribution of historical Oncomelania hupensis snail environments and wetland environments in Wuxi City].

OBJECTIVE: To understand the distribution and characteristics of historical Oncomelania hupensis snail environments and wetland environments in Wuxi City since 1952, so as to provide the evidence for formulating the monitoring strategy after the schistosomiasis transmission was interrupted. METHODS: A cross-sectional survey was conducted to collect the information of historical O. hupensis snail environments and wetland environments. Google Earth was used to present and analyze the distribution and characteristics of them. RESULTS: There were 2 124 historical O. hupensis snail environments in Wuxi City and the accumulative area was 2 995.7 hm2. In 2013, there were 61 surveillance sites of wetland with the area of 32.8 hm2; totally 1 695 snails were caught and no O. hupensis snails were found. CONCLUSION: The historical O. hupensis snail environments were distributed widely and covered a large area in Wuxi City. The surveillance should be focused on the key environments and wetland.

[Study on the HIV infection status, characteristics of AIDS-related behaviors and risk factors among men attending the sexually transmitted disease clinics].

OBJECTIVE: To understand the status on AIDS awareness, AIDS-related behaviors, risk factors on HIV infection status among 15 - 90 years or older men at the sexually transmitted disease clinics. METHODS: Data from the 2009 and 2010 national sentinel surveillance system, regarding men attending sexually transmitted disease clinics was collected from Guangdong, Guangxi, Henan, Sichuan, Yunnan and Jiangxi provinces, where the AIDS epidemic among 15 - 90 years or older population was serious. Data was uploaded to National Center for AIDS/STD Control and Prevention (NCAIDS) through the comprehensive AIDS control and prevention information system. Data was then analyzed by SPSS 18.0. RESULTS: A total of 64 003 pieces of data were collected. Among them, 8783 (13.7%) were related to men at ≥ 50 years or older, and to men 15 - 49 years older were 55 220. The rates on the awareness of AIDS knowledge were from 69.6% vs. 80.1%, on frequently having had commercial sexual contacts in the last three months were between 34.1% vs. 36.6%, on having had casual sexual contact in the last three months were 18.7% vs. 28.4%, on having had homosexual anal intercourse as 0.7% vs. 1.4%. The rates of taking HIV antibody testing in the last year (14.3% vs. 17.1%) among this population were all significantly lower than the rate among the 15 to 49 years age group. However, the HIV-positive rate among the older age group (fifty years of age or older) was significantly higher than the rate among 15 to 49 year age group (1.1% vs. 0.7%). Regard the fifty years of age or older men. Data from the multivariate logistic regression analysis demonstrated that factors as: having regular partner (OR = 0.588, P = 0.034), having homosexual anal intercourse (OR = 5.226, P = 0.006) were associated with positivities of HIV antibody. CONCLUSION: High-risk sexual behaviors, including homosexual anal intercourse were the major risk factors for men at ≥ 50 years or older age, related to the infection of HIV.