Suberoylanilide hydroxamic acid upregulates reticulophagy receptor expression and promotes cell death in hepatocellular carcinoma cells

BACKGROUND Hepatocellular carcinoma(HCC)is a common clinical condition with a poor prognosis and few effective treatment options.Potent anticancer agents for treating HCC must be identified.Epigenetics plays an essential role in HCC tumorigenesis.Suberoylanilide hydroxamic acid(SAHA),the most common histone deacetylase inhibitor agent,triggers many forms of cell death in HCC.However,the underlying mechanism of action remains unclear.Family with sequence similarity 134 member B(FAM134B)-induced reticulophagy,a selective autophagic pathway,participates in the decision of cell fate and exhibits anticancer activity.This study focused on the relationship between FAM134B-induced reticulophagy and SAHA-mediated cell death.AIM To elucidate potential roles and underlying molecular mechanisms of reticulophagy in SAHA-induced HCC cell death.METHODS The viability,apoptosis,cell cycle,migration,and invasion of SAHA-treated Huh7 and MHCC97L cells were measured.Proteins related to the reticulophagy pathway,mitochondria-endoplasmic reticulum(ER)contact sites,intrinsic mitochondrial apoptosis,and histone acetylation were quantified using western blotting.ER and lysosome colocalization,and mitochondrial Ca^(2+)levels were characterized via confocal microscopy.The level of cell death was evaluated through Hoechst 33342 staining and propidium iodide colocalization.Chromatin immunoprecipitation was used to verify histone H4 lysine-16 acetylation in the FAM134B promoter region.RESULTS After SAHA treatment,the proliferation of Huh7 and MHCC97L cells was significantly inhibited,and the migration and invasion abilities were greatly blocked in vitro.This promoted apoptosis and caused G1 phase cells to increase in a concentration-dependent manner.Following treatment with SAHA,ER-phagy was activated,thereby triggering autophagy-mediated cell death of HCC cells in vitro.Western blotting and chromatin immunoprecipitation assays confirmed that SAHA regulated FAM134B expression by enhancing the histone H4 lysine-16 acetylation in the FAM134B promoter region.Further,SAHA disturbed the Ca^(2+)homeostasis and upregulated the level of autocrine motility factor receptor and proteins related to mitochondria-endoplasmic reticulum contact sites in HCC cells.Additionally,SAHA decreased the mitochondrial membrane potential levels,thereby accelerating the activation of the reticulophagy-mediated mitochondrial apoptosis pathway and promoting HCC cell death in vitro.CONCLUSION SAHA stimulates FAM134B-mediated ER-phagy to synergistically enhance the mitochondrial apoptotic pathway,thereby enhancing HCC cell death.

Analysis of In-hospital Neonatal Death in the Tertiary Neonatal Intensive Care Unit in China： A Multicenter Retrospective Study

Background：Globally,the proportion of child deaths that occur in the neonatal period remains a high level of 37-41%.Differences of cause in neonate death exist in different regions as well as in different economic development countries.The specific aim of this study was to investigate the causes,characteristics,and differences of death in neonates during hospitalization in the tertiary Neonatal Intensive Care Unit （NICU） of China.Methods：All the dead neonates admitted to 26 NICUs were included between January 1,2011,and December 31,2011.All the data were collected retrospectively from clinical records by a designed questionnaire.Data collected from each NICU were delivered to the leading institution where the results were analyzed.Results：A total of 744 newborns died during the l-year survey,accounting for 1.2% of all the neonates admitted to 26 NICUs and 37.6% of all the deaths in children under 5 years of age in these hospitals.Preterm neonate death accounted for 59.3% of all the death.The leading causes of death in preterm and term infants were pulmonary disease and infection,respectively.In early neonate period,pulmonary diseases （56.5%） occupied the largest proportion ofpreterm deaths while infection （27%） and neurologic diseases （22%） were the two main causes of term deaths.In late neonate period,infection was the leading cause of both preterm and term neonate deaths.About two-thirds of neonate death occurred after medical care withdrawal.Of the cases who might survive if receiving continuing treatment,parents＇ concern about the long-term outcomes was the main reason of medical care withdrawal.Conclusions：Neonate death still accounts for a high proportion of all the deaths in children under 5 years of age.Our study showed the majority of neonate death occurred in preterm infants.Cause of death varied with the age of death and gestational age.Accurate and prompt evaluation of the long-term outcomes should be carried out to guide the critical decision.

Dwell time and bloodstream infection incidence of umbilical venous catheterization in China

Importance:Central line-associated bloodstream infection(CLABSI)is one of the most serious complications of central venous access devices.Reducing the risk of CLABSI is of utmost significance in efforts to improve neonatal mortality rates and enhance long-term prognosis.Objective:To determine the dwell time and incidence of CLABSI of umbilical venous catheterization(UVC)for preterm infants in China.Methods:Preterm infants with UVC admitted to 44 tertiary neonatal intensive care units in 24 provinces in China were enrolled.Study period was from November 2019 to August 2021.The end point of observations was 48 h after umbilical venous(UV)catheter removal.The primary outcomes were dwell time of UV catheter and UVC-associated CLABSI.Data between infants with UV catheter dwell time≤7 days and>7 days,and with birth weight(BW)≤1000 g and>1000 g were compared.Results:In total,2172 neonates were enrolled(gestational age 30.0±2.4 weeks,BW 1258.5±392.8 g).The median UV catheter dwell time was 7(6–10)days.The incidence of UVC-associated CLABSI was 3.03/1000 UV catheter days.For infants with UV catheter dwell time≤7 days and>7 days,the UVC-associated CLABSI incidence was 3.71 and 2.65 per 1000 UV catheter days,respectively,P=0.23.For infants with UVC dwell times of 3–6,7–12,and 13–15 days,the UVC-associated CLABSI rates were 0.14%,0.68%,and 2.48%(P<0.01).The Kaplan–Meier plot of UV catheter dwell time to CLABSI showed no difference between infants with BW≤1000 g and>1000 g(P=0.60).Interpretation:The median dwell time of UV catheter was 7 days,and the incidence of UVC-associated CLABSI was 3.03/1000 catheter days in China.The daily risk of UVC-associated CLABSI and other complications increased with the dwell time.

Mortality and Morbidity of Extremely Low Birth Weight Infants in the Mainland of China： A Multi-center Study

Background： With the progress ofperinatal medicine and neonatal technology, more and more extremely low birth weight （ELBW） survived all over the world. This study was designed to investigate the short-term ontcomes of ELBW infants during their Neonatal Intensive Care Unit （NICU） stay in the mainland of China. Methods： All infants admitted to 26 NICUs with a birth weight （BW） 〈1000 g were included between January 1,2011 and December 31,2011. All the data were collected retrospectively from clinical records by a prospectively designed questionnaire. The data collected from each NICU transmitted to the main institution where the results were aggregated and analyzed. Categorical variables were performed with Pearson Chi-square test. Binary Logistic regression analysis was used to detect risk factors. Results： A total of 258 ELBW infants were admitted to 26 NICUs, of whom the mean gestational age （GA） was 28.1 ± 2.2 weeks, and the mean BW was 868 ± 97 g. The overall survival rate at discharge was 50.0%. Despite aggressive treatment 60 infants （23.3%） died and another 69 infants （26.7%） died after medical care withdrawal. Furthermore, the survival rate was significantly higher in coastal areas than inland areas （53.6% vs. 35.3%, P = 0.019）. BW 〈750 g and GA 〈28 weeks were the largest risk factors, and being small for gestational age was a protective factor related to mortality. Respiratory distress syndrome was the most common complication. The incidence of patent ductus arteriosus, intraventricular hemorrhage, periventricular leukomalacia, bronchopulmonary dysplasia, retinopathy of prematurity was 26.2%, 33.7%, 6.7%, 48.1%, and 41.4%, respectively. Ventilator associated pneumonia was the most common hospital acquired infection during hospitalization. Conclusions： Our study was the first survey that revealed the present status of ELBW infants in the mainland of China. The mortality and morbidity of ELBW infants remained high as compared to other developed countries.