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 Ca2+ 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 Ca2+ 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.

Glucagon-Like Peptide-2 Ameliorates Age-Associated Bone Loss and Gut Barrier Dysfunction in Senescence-Accelerated Mouse Prone 6 Mice.

INTRODUCTION: Senile osteoporosis is one of the most common age-related diseases worldwide. Glucagon like peptide-2 (GLP-2), a naturally occurring gastrointestinal peptide, possesses therapeutic effects on bone loss in postmenopausal women and ovariectomized rats. However, the role of GLP-2 in senile osteoporosis and underlying mechanisms has not been explored. METHODS: GLP-2 was subcutaneously injected into the 6-month-old male senile osteoporosis model of senescence-accelerated mouse prone 6 (SAMP6) mice for 6 weeks. SAMP6 subjected to normal saline and senescence-accelerated mouse resistant 1 served as control groups. Micro-computed tomography was performed to evaluate the bone mass and microarchitecture of the mice. Osteoblastic and osteoclastic activities were determined by biochemical, quantitative real-time PCR, histological, and histomorphometric analyses combined with hematoxylin-eosin, toluidine blue, and tartrate-resistant acid phosphatase staining. We also examined the proteins and structure of intestinal tight junction using immunohistochemical assay as well as a transmission electron microscope. Serum inflammation marker levels were measured using ELISA. Additionally, anti-oxidative enzymes GPX-4 and SOD-2 and receptors of GLP-2 and vitamin D expression in the ileum and colon were detected under immunofluorescence staining. RESULTS: Six-week GLP-2 treatment attenuated bone loss in SAMP6 mice, as evidenced by increased bone mineral density, improved microarchitecture in femora, and enhanced osteogenic activities. In contrast, the activity of osteoclastic activity was not obviously inhibited. Moreover, GLP-2 ameliorated tight junction structure and protein expression in the intestinal barrier, which was accompanied by the reduction of TNF-α level. The expression of receptors of intestinal GLP-2 and vitamin D in the ileum was elevated. Furthermore, the oxidative stress in the intestines was improved by increasing the GPX-4 and SOD-2 signaling. CONCLUSION: Our findings suggest that GLP-2 could ameliorate age-associated bone loss, tight junction structure, and improved antioxidant enzyme activity in the gut in SAMP6 mice. Amelioration of gut barrier dysfunction may potentially contribute to improving bone formation and provide evidence for targeting the entero-bone axis in the treatment of senile osteoporosis.

Different Effects of Antibiotics on Klebsiella pneumoniae and Escherichia coli Resistance Induced by Antibiotics: A Retrospective Study from China.

Objective: The main objective was to assess the correlation between antibiotic use and carbapenem-resistant Klebsiella pneumoniae (CRKP) and carbapenem-resistant Escherichia coli (CREC) induction by antibiotics. Materials and Methods: A retrospective cohort study was conducted from January 2017 to December 2020. This study included patients with K. pneumoniae and E. coli. Kaplan-Meier analysis and Cox proportional hazard model were used to estimate the hazard of carbapenem-resistant Enterobacterales (CRE), whereas restricted cubic spline regression was used to visualize the hazard of CRE by antibiotics at different doses. Results: Two thousand fifty-six K. pneumoniae patients and 3,243 E. coli patients were included. After Cox proportional hazard model analysis, carbapenems or 1st-cephalospoins or penicillin monotherapy, male and ICU admission were associated with CRKP. CREC was associated with quinolone monotherapy. Time-to-event analysis indicated that carbapenem, ß-lactamase inhibitor mixtures, and quinolones were associated with higher 30-day CRKP hazards than other antibiotics (χ2 = 33.670, p < 0.001). Further restricted cubic spline regression analysis found that the hazard of CRKP induction decreased with the increased dose of ß-lactamase inhibitor mixtures, but there was no significant change in the hazard ratio of CRKP induction with the increased dose of quinolones. Moreover, there was an obvious characteristic of "parabolic curve" for the hazard of CREC induction due to ß-lactamase inhibitor mixtures, and the hazard value gradually increased with the dose, reached the maximum at 24 g, and finally gradually decreased from 26 g. Conclusions: Rational use of antibiotics should be implemented and antimicrobial stewardship policies should be adjusted according to the characteristics of each hospital.

A facile access to N-sulfonylthioimidates and their use for the transformation to 3,4-dihydroquinazolines.

N-Sulfonylthioimidates can be efficiently synthesized through one-pot three-component coupling of terminal alkynes, sulfonyl azides, and thiols by using a copper(i) catalyst in the presence of 4-dimethylaminopyridine. The proposed reaction is characterized by mild reaction conditions and tolerance of diverse functional groups. Additionally, the crucial pharmacophore of 3,4-dihydroquinazolines was synthesized using a one-pot synthetic strategy from N-sulfonylthioimidates.

Highly sensitive serological approaches for Pepino mosaic virus detection.

Pepino mosaic virus (PepMV) causes severe disease in tomato and other Solanaceous crops around globe. To effectively study and manage this viral disease, researchers need new, sensitive, and high-throughput approaches for viral detection. In this study, we purified PepMV particles from the infected Nicotiana benthamiana plants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies (mAbs). A panel of highly specific and sensitive murine mAbs (15B2, 8H6, 23D11, 20D9, 3A6, and 8E3) could be produced through cell fusion, antibody selection, and cell cloning. Using the mAbs as the detection antibodies, we established double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), Dot-ELISA, and Tissue print-ELISA for detecting PepMV infection in tomato plants. Resulting data on sensitivity analysis assays showed that both DAS-ELISA and Dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1 310 720 and 1:20 480 (weight/volume ratio (w/v), g/mL), respectively. Among the three methods developed, the Tissue print-ELISA was found to be the most practical detection technique. Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction (RT-PCR) and DNA sequencing, demonstrating all three serological methods are reliable and effective for monitoring PepMV. Anti-PepMV mAbs and the newly developed DAS-ELISA, Dot-ELISA, and Tissue print-ELISA can benefit PepMV detection and field epidemiological study, and management of this viral disease, which is already widespread in tomato plants in Yunnan Province of China.

Quarantine Vehicle Scheduling for Transferring High-Risk Individuals in Epidemic Areas.

In a large-scale epidemic outbreak, there can be many high-risk individuals to be transferred for medical isolation in epidemic areas. Typically, the individuals are scattered across different locations, and available quarantine vehicles are limited. Therefore, it is challenging to efficiently schedule the vehicles to transfer the individuals to isolated regions to control the spread of the epidemic. In this paper, we formulate such a quarantine vehicle scheduling problem for high-risk individual transfer, which is more difficult than most well-known vehicle routing problems. To efficiently solve this problem, we propose a hybrid algorithm based on the water wave optimization (WWO) metaheuristic and neighborhood search. The metaheuristic uses a small population to rapidly explore the solution space, and the neighborhood search uses a gradual strategy to improve the solution accuracy. Computational results demonstrate that the proposed algorithm significantly outperforms several existing algorithms and obtains high-quality solutions on real-world problem instances for high-risk individual transfer in Hangzhou, China, during the peak period of the novel coronavirus pneumonia (COVID-19).

Predictors of primary patency after percutaneous balloon angioplasty for stenosis of Brescia-Cimino hemodialysis arteriovenous fistula.

OBJECTIVE: Percutaneous transluminal balloon angioplasty (PTA) is recommended as the first choice to treat stenosis of Brescia-Cimino arteriovenous fistulas (B-C AVFs). The ability to predict which B-C AVFs are at risk for recurrent stenosis post-PTA would allow closer monitoring of patients, and possibly result in surgical intervention rather than repeat PTA. The purpose of this study was to identify predictive factors of primary patency after PTA in B-C AVFs. METHODS: Patients diagnosed with B-C AVF primary stenosis and treated by PTA between November 2013 and March 2018 were included in the study. Patient and stenotic lesion characteristics and PTA procedure factors were included in the analysis. The Kaplan-Meier method was used to analyze the primary patency rate. Cox proportional hazard regression analysis was used to identify factors predictive of decreased primary patency. RESULTS: 74 patients (35 males, 39 females) with a mean age of 61.68 ± 11.44 years (range, 36-84 years) were included in the study. The mean B-C AVF age was 16.34 ± 12.93 months (range, 2-84 months), and the median primary patency time was 7.79 ± 0.48 months. Cox proportional hazard regression analysis revealed stenosis location at the inflow artery [hazard ratio (HR)=3.83, 95% confidence interval (CI): 1.46-10.09] or anastomosis (HR = 1.90, 95% CI: 1.09-3.32), dilation >2 times during PTA (HR = 2.30, 95% CI: 1.22-4.34), and residual stenosis >30% (HR = 2.42, 95% CI: 1.26-4.63) were significantly associated with decreased patency. CONCLUSION: In conclusion, the primary patency rate of PTA for B-C AVF dysfunction is reduced by dilation >2 times, residual stenosis >30%, and stenosis located at the inflow artery or anastomosis. These results may help in tailoring surveillance programs, multiple PTA, or a proximal re-anastomosis surgery in patients with AVF dysfunction. ADVANCES IN KNOWLEDGE: A number of studies have been conducted to examine the predictors of primary patency after PTA, however, no definitive conclusions have been reached. Our study revealed that stenosis location at the inflow artery or anastomosis, dilation >2 times during PTA, and residual stenosis >30% were the predictors of primary patency after PTA, which may help in tailoring surveillance programs, multiple PTA, or a proximal re-anastomosis surgery in patients with arteriovenous fistulas dysfunction.

A multi-center nested case-control study on hospitalization costs and length of stay due to healthcare-associated infection.

Background: In 2018, the Chinese government demanded nationwide implementation of medical insurance payment methods based on Single-Disease Payment (SDP), but during the operation process the medical insurance system did not fully consider the extra economic burden caused by healthcare-associated infection (HAI). HAIs can prolong the length of stay and increase the hospitalization costs, but only a few studies have been conducted in Sichuan province, China. We evaluated the hospitalization costs and length of stay due to HAI in Sichuan province based on the prevalence survey, and provided data reference for China's medical insurance reform. Methods: In the hospitals surveyed on the prevalence of HAI, a multi-center nested case-control study was performed by a paired method. The study period was from 6 September 2016 to 30 November 2016. Binary outcomes were tested using χ2 test, continuous outcomes were tested using Wilcoxon matched-pairs signed rank test, intra-group comparisons were tested using multiple linear regression analysis. Results: A total of 225 pairs/450 patients were selected in 51 hospitals, and 170 pairs/350 patients were successfully matched. The case fatality rate was 5.14% for the HAIs patients and 3.43% for non-HAs patients, there was no significant difference (χ2 = 0.627, P = 0.429); the median length of stay in patients with HAIs was 21 days, longer than that of patients with non-HAI 16 days, the median of the difference between matched-pairs was 5 days, the difference was statistically significant (Z = 4.896, P = 0.000). The median hospitalization costs of patients with HAI were €1732.83, higher than that of patients with non-HAI €1095.29, the median of the difference between matched-pairs were €431.34, the difference was statistically significant (Z = 6.413, P = 0.000). Multiple linear regression results showed that HAIs at different sites have caused different economic burdens, but in different economic regions, the difference was not statistically significant. Conclusions: In Sichuan, the hospitalization costs and length of stay caused by HAI should be given special attention in the current medical insurance reform. The proportion and scope of medical payment for patients with HAI at different sites should be different. Efforts need to be taken to incentivize reduction of HAI rates which will reduce hospitalization costs and length of stay.

Monoclonal antibody-based serological assays for detection of Potato virus S in potato plants.

Potato virus S (PVS) often causes significant losses in potato production in potato-growing countries. In this study, the ordinary strain of PVS (PVSO) was purified from PVS-infected potato plants and used as the immunogen to produce hybridomas secreting monoclonal antibodies (MAbs). Five highly specific and sensitive murine MAbs (1A3, 16C10, 18A9, 20B12, and 22H4) against PVS were prepared using conventional hybridoma technology. Using these MAbs, tissue print-enzyme-linked immunosorbent assay (ELISA), dot-ELISA, and double-antibody sandwich (DAS)-ELISA were developed for sensitive and specific detection of PVS infection in potato plants. The results of sensitivity assays revealed that PVS could be reliably detected in PVS-infected leaf crude extracts diluted at 1:10 240 and 1:163 840 (w/v, g/ml) in phosphate buffer saline (PBS) by dot-ELISA and DAS-ELISA, respectively. Twenty-two samples collected from potato fields in Yunnan Province, China were tested for PVS infection using the serological assays we had developed, and 14 of them were found to be positive. This indicates that PVS is now prevalent in potato fields in Yunnan Province.

Photo-Fenton effect of 4-chlorophenol in ice.

The photoconversion of 4-chlorophenol (4-CP) in a simulated sunlight-Fenton system was investigated in ice and aqueous solution. It was found that the hydroxyl radical (OH) had an important effect on the photoconversion of 4-CP in both phases, but the effects of Cl(-), SO4(2-), NO3(-), and HCO3(-) were different. In aqueous solution, the photoconversion efficiency of 4-CP was proportional to the OH concentration, and hence, Cl(-) and HCO3(-) as OH scavengers prohibited the photoconversion; SO4(2-) had little effect; NO3(-) promoted the process under certain conditions owing to OH being generated by the photolysis of NO3(-). In ice, however, the photoconversion efficiency of 4-CP was not proportional to the concentration of OH. The photoconversion efficiency of 4-CP increased with increasing concentrations of all ions, although the OH remained almost constant, only increasing or decreasing slightly. This provides new evidence for the presence of a quasi-liquid layer (QLL). Hydroxylation products were detected in both phases. All photoproducts in aqueous solution contained only a single benzene ring, whereas in ice, dimers were also detected.

[Changes of erythrocyte rheology in rabbits with acute renal failure].

OBJECTIVE: To observe the change of erythrocyte theology in rabbits with acute renal failure (ARF). METHODS: Thirty-eight healthy rabbits were randomly divided into control group (n = 8), model group (establishing ARF model via intramuscular injection 1% HgCl2, and divided into 12 h, 24 h, 48 h subgroups, all n = 10), the arterial blood sample was taken out through carotid artery at corresponding times after anesthetization with urethane, for detecting the indices of renal function and erythrocyte rheology. RESULTS: The levels of urea and creatinine in plasma of model rabbits at 12 h, 24 h and 48 h were higher than those of control group, and there was a rise trend along with the time extension. The erythrocytes electrophoresis time at 12 h of model group was higher, the electrophoresis rate and migration rate of erythrocytes were lower compared with those of control group; the erythrocytes electrophoresis time at 24 h of model group was lower and the electrophoresis rate and migration rate were higher compared with those of model group at 12 h; and there were no statistical differences in erythrocytes electrophoresis indices between model group at 48 h and other groups. Meanwhile, there was a rise trend in erythrocyte sedimentation rate (ESR), K value of equation and emendation along with the time extension of ARF, but these indices only at 48 h of model group was lower significantly than that of control group. There were no statistical differences in aggregation index and deformability index of erythrocytes among groups. CONCLUSION: During the process of ARF, the erythrocytes electrophoresis time lengthen and electrophoresis rate and migration rate decrease at early stage, and these indices gradually return to normal; the indices of ESR increase gradually.

[Establishment of in vitro co-cultue system for tumor vascular endothelial cells].

OBJECTIVE: To establish a in vitro culture model of tumor vascular endothelial cells (TECs). METHODS: We established in vitro culture model of TECs, named A549-HUVECs, through the co-culutre system culturing primary human umbilical vein endothelial cells (HUVECs) and human lung adenocarcinoma epithelial cell line A549 cells in indirect co-cultures, six-well Transwell-clear dishes (0.4 microm porous filter, Costar). We then observed TECs' cell morphology, growth characteristics, proliferation and karyotype, telomerase activity, etc. RESULTS: In the indirect co-culture system, A549-HUVECs showed migration cell-type appearance, more active cellular metabolism, increasing cell permeability, augmenting cell proliferation, which indicated A549-HUVECs being in a state of the regulation of homeostasis imbalance for cell proliferation and apoptosis. Besides, the analysis of A549-HUVECs genetic characteristics showed that the chromosome karyotype of A549-HUVECs was sub-triploid, and its telomerase activity increased, phenotypic change occurred, which possessed the similar pathological features of precancerous lesion. CONCLUSION: A549-HUVECs co-culture system could be feasible for tumor vascular research.

Dyslipidemia in Shanghai, China.

OBJECTIVES: To determine the prevalence of and risk factors for dyslipidemia in Shanghai. METHODS: A cross-sectional survey of 14,385 subjects (6150 men) with mean age of 49.5 (14.5) years was conducted between October 2002 and April 2003 using randomized, stratified cluster sampling. Serum triglyceride, total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) were measured. RESULTS: Dyslipidemia, as defined by NCEP ATP III criteria, occurred in 5255 subjects (36.5%). The prevalences of mixed hyperlipidemia (elevated TC and triglycerides), isolated hypertriglyceridemia, isolated hypercholesterolemia and isolated low HDL-C were 3.8%, 24.9%, 3.2% and 4.7%, respectively. The prevalence of dyslipidemia increased with age, with the peak prevalence (43%) occurring after age 55. Dyslipidemia was more common in males than females (40.2% vs. 33.8%) and in rural than urban populations (44.2% vs. 32.3%). Serum triglyceride and TC increased with body mass index (BMI) and waist circumference. Mean serum triglyceride concentrations in males and rural residents were higher than those in females and urban residents, respectively, whereas the reverse was true for HDL-C values. Multivariate analysis revealed that dyslipidemia was associated with age, gender, area of residence, BMI and waist circumference. CONCLUSIONS: There is a high prevalence of dyslipidemia, mainly hypertriglyceridemia and low HDL-C, in Shanghai.

[Microbiology and management of gaseous gangrene:the Sichuan experience of 20 earthquake cases].

Wound secretion from 20 patients with gaseous gangrene was collected for Gram staining, bacterial culture and drug sensitivity tests. The results indicated that gaseous gangrene was caused by the co-infection of both aerobic and anaerobic bacteria. Gram-negative bacilli were slightly more common than other aerobic bacteria in gaseous gangrene wound, which was different from the findings of ordinary gaseous gangrene.