Search

Abstract Background Alcohol abuse is involved in the pathogenesis of multiple organ disorders; the underlying mechanism is incompletely understood. The ubiquitin editing enzyme A20 is involved in regulating activities in the cell. Suppression of A20 is suggested as one factor in the initiation of inflammation. This study investigates the mechanism by which chronic alcohol consumption modulates the levels of ubiquitin editing enzyme A20 in macrophages and further contributes to induce endothelial barrier dysfunction in the lung. Methods Mice were gavage-fed with 40% alcohol daily for 0-3 weeks. Airway macrophages were collected by lung lavage. Expression of ubiquitin editing enzyme A20 in isolated macrophages was assessed at both mRNA and protein levels. The endothelial barrier function of the lung was evaluated by the Evans blue method. Results Mice treated with alcohol for 3 weeks showed an increase in cell infiltration in the lung in response to exposure to peptidoglycan; over 80% of the infiltrated cells were macrophages. Furthermore, we observed that A20 level was suppressed in macrophages of mice treated with alcohol; the levels of tumor necrosis factor, interleukin-6 and nuclear factor kappa B in macrophage were increased. In addition, the endothelial barrier function of the lung was compromised, showing excessive infiltration of Evans blue in the lung indicating lung edema. Pretreatment with synthesized A20 inhibited alcohol-induced lung endothelial barrier dysfunction. Conclusions We conclude that chronic alcohol ingestion disturbs the endothelial barrier function in the lung by modulating macrophage properties. Increase in A20 in the cell may have potential for the treatment of inflammatory disorders.

Objective To discuss the methods and strategies to identify the causes of dependents' deaths, as well as provide the experiences that can be used for reference and scientific basis for the forensic identification of the potentially growing deaths of the same kind in the future. Methods The 13 cases concerning death of dependents accepted by Sun Yat-sen University Forensic Center were collected, and the basic information of the dependents were statistically described. The nutritional status, environmental condition and medical care condition were evaluated according to dietary energy, living space, environment and medical treatment condition. Results Among the 13 dependents, there were 11 males and 2 females, with the oldest 74 and the youngest 9 and dwelling time was from 0.4 to 5.6 years. Forensic pathological examination showed that 13 dependents had infectious diseases and 11 were severely dystrophic. There were no fatal mechanical injuries or poisoning in dependents. Molecular pathological screening of 4 cases revealed no pathogenic variants of sudden death susceptible genes. The poor status of the diet, nutrition, living environment and medical care of these dependents were discovered. The direct cause of death of all 13 dependents was identified to be disease. The lack of nutrition, poor living environment and lack of medical care were thought to play a dominant role in causing the deaths of 12 dependants. Conclusion The death identification should follow the judicial procedure. In identification of the causes of death and analysis of the proportion of the affecting factors resulting in death, all factors, including nutrition, environment, medical care, injury and diseases, need to be considered. [ABSTRACT FROM AUTHOR]

Antibiotic-producing microorganisms have evolved several self-resistance mechanisms to prevent auto-toxicity. Overexpression of specific transporters to improve the efflux of toxic antibiotics has been found one of the most important and intrinsic resistance strategies used by many Streptomyces strains. In this work, two ATP-binding cassette (ABC) transporter-encoding genes located in the natamycin biosynthetic gene cluster, scnA and scnB, were identified as the primary exporter genes for natamycin efflux in Streptomyces chattanoogensis L10. Two other transporters located outside the cluster, a major facilitator superfamily transporter Mfs1 and an ABC transporter NepI/II were found to play a complementary role in natamycin efflux. ScnA/ScnB and Mfs1 also participate in exporting the immediate precursor of natamycin, 4,5-de-epoxynatamycin, which is more toxic to S. chattanoogensis L10 than natamycin. As the major complementary exporter for natamycin efflux, Mfs1 is up-regulated in response to intracellular accumulation of natamycin and 4,5-de-epoxynatamycin, suggesting a key role in the stress response for self-resistance. This article discusses a novel antibiotic-related efflux and response system in Streptomyces, as well as a self-resistance mechanism in antibiotic-producing strains. [ABSTRACT FROM AUTHOR]

Background: Cardiogenic stroke is associated with substantial morbidity and mortality, necessitating a better understanding of its clinical characteristics for improved patient outcomes. This study aimed to identify clinical characteristics influencing short-term functional prognosis in patients with cardiogenic stroke., Methods: The study prospectively enrolled 212 patients with cardiogenic stroke, collecting their clinical data and laboratory results. The modified Rankin Scale score at 90 days was used to define functional prognosis, with patients having a good prognosis (modified Rankin Scale ≤2; n = 164) or poor prognosis (modified Rankin Scale ≥3; n = 48)., Results: The poor prognosis group had higher rates of total anterior circulation infarcts (12.5% vs 0.0%; P < .001) and posterior circulation infarction (50.0% vs 38.4%; P < .001) compared with the good prognosis group. Lesion characteristics differed significantly, with the poor prognosis group exhibiting more large-area lesions (39.6% vs 18.9%; P < .001) and multiple confluent lesions (56.3% vs 24.4%; P < .001). Admission-based National Institute of Health Stroke Scale scores were higher in the poor prognosis group (median [IQR], 12 [8-18] vs 5 [4-7]; P <.001), correlating with worse outcomes. The admission National Institute of Health Stroke Scale score predicted patients' 90-day prognosis with good accuracy (area under the curve, 0.937 [95% CI, 0.895-0.965]; P < .001), with a threshold of 7 yielding 85.42% sensitivity and 85.37% specificity., Conclusion: Higher admission National Institute of Health Stroke Scale scores were significantly associated with poor functional prognosis at 90 days, highlighting the importance of early National Institute of Health Stroke Scale-based assessment for improved outcomes., Competing Interests: Conflict of Interest Disclosure: The authors declare that they have no competing interests., (© 2024 The Authors. Published by The Texas Heart Institute®.)

Hepatocellular carcinoma (HCC) is the most prevalent primary liver cancer in adults. Previous studies in our laboratory found that long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) was upregulated in HCC cells, which could affect the metastasis and invasion of HCC. However, the underlying mechanism remains unknown. Herein, we studied the interaction between MALAT1 and miR-140 on the regulation of angiogenesis and immunosuppressive properties. We revealed that the expression of MALAT1 and VEGF-A was significantly increased in HCC cells. Knockdown of MALAT1 in HCC cells suppressed the production of VEGF-A, impaired the angiogenesis of HUVECs, and facilitated the polarization of macrophage toward the M1 subset. Mechanistically, the interaction between MALAT1 and miR-140 or between miR-140 and VEGF-A was confirmed by multiple assays. Besides, a negative correlation between MALAT1 and miR-140 was found in HCC tissues. Furthermore, miR-140 inhibition significantly increased VEGF-A expression, promoted angiogenesis of HUVECs, and redirected the polarization of macrophages toward the M2 subset. In addition, in vivo studies also verified the regulatory network of the MALAT1/miR-140 axis on VEGF-A in HCC progression. In summary, this study revealed the mechanism that MALAT1 worked as a putative HCC promotor via inhibiting miR-140. Therefore, targeting MALAT1 or miR-140 might alleviate the progression of HCC in the future.

Detailed mechanisms of WhiB-like (Wbl) proteins involved in antibiotic biosynthesis and morphological differentiation are poorly understood. Here, we characterize the role of WblAch, a Streptomyces chattanoogensis L10 protein belonging to this superfamily. Based on DNA microarray data and verified by real-time quantitative PCR (qRT-PCR), the expression of wblAch was shown to be positively regulated by AdpAch. Gel retardation assays and DNase I footprinting experiments showed that AdpAch has specific DNA-binding activity for the promoter region of wblAch. Gene disruption and genetic complementation revealed that WblAch acts in a positive manner to regulate natamycin production. When wblAch was overexpressed in the wild-type strain, the natamycin yield was increased by ∼30%. This provides a strategy to generate improved strains for natamycin production. Moreover, transcriptional analysis showed that the expression levels of whi genes (including whiA, whiB, whiH, and whiI) were severely depressed in the ΔwblAch mutant, suggesting that WblAch plays a part in morphological differentiation by influencing the expression of the whi genes., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

Objective: To explore the application value of serum total IgE, tryptase and chymase in the identification of death caused by drug anaphylactic shock., Methods: The general information from 235 cases of non-drug anaphylactic shock and 32 cases of drug anaphylactic shock were analyzed. The serum IgE level had been detected in the cases. Ten cases caused by coronary disease and 10 cases caused by sudden manhood death syndrome were selected from non-drug anaphylactic shock cases for the control group. Expressions of tryptase and chymase in the lung and heart were detected using immunohistochemistry method. The number and IOD of positive mast cells were counted., Results: In the drug anaphylactic shock group, the IgE value of 18 samples (56.25%) was significantly higher than the normal upper limit of 120 IU/mL. In the non-drug anaphylactic shock group, the IgE value of 67 samples (28.51%) was higher than 120 IU/mL. The expressions of tryptase and chymase were significantly increased in lung and myocardial tissue in drug anaphylactic shock group (P < 0.05)., Conclusion: Tryptase and chymase are more superior than that of the serum total IgE in the diagnosis of death caused by drug anaphylactic shock, and are more suitable in forensic practice.

Dexamethasone (DEX)-induced heart damage is associated with enzyme and protein alterations. The purpose of this study was to investigate DEX-induced alterations in cardiac enolase and caveolin-1 (cav-1) following DEX administration. Male Wistar rats were randomly divided into two groups: a control and a DEX. The DEX group intraperitoneally received DEX at the single dose of 10 mg/kg for 7 consecutive days, and the control was given the same amount of saline via the same route. On day 8, the rats were anesthetized, and a thoracotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of enolase and cav-1. Sections were analyzed by digital image analysis. Our results demonstrated that cardiac protein expression of enolase and cav-1 was altered following DEX-induced toxicity in the rat. The expression of enolase and cav-1 was significantly increased after DEX treatment, supported by integrated optical density compared with the control (P<0.05). In conclusion, following DEX-induced toxicity, protein expression of enolase and cav-1 was significantly elevated. The current findings indicate that such alterations would be reflected in abnormal cardiac function, and the proteins identified in this study may be useful in revealing the mechanisms underlying DEX-induced toxicity and also in providing various clues for further research.

Objective: To explore the mechanism that how human enterovirus 71 (EV71) invades the brainstem and how intercellular adhesion molecules-1 (ICAM-1) participates by analyzing the expression and distribution of human EV71, and ICAM-1 in brainstem of infants with brain stem encephalitis., Methods: Twenty-two brainstem of infants with brain stem encephalitis were collected as the experimental group and 10 brainstems of fatal congenital heart disease were selected as the control group. The sections with perivascular cuffings were selected to observe EV71-VP1 expression by immunohistochemistry method and ICAM-1 expression was detected for the sections with EV71-VP1 positive expression. The staining image analysis and statistics analysis were performed. The experiment and control groups were compared., Results: (1) EV71-VP1 positive cells in the experimental group were mainly astrocytes in brainstem with [dark]-brown particles, and the control group was negative. (2) ICAM-1 positive cells showed [dark]-brown. The expression in inflammatory cells (around blood vessels of brain stem and in glial nodules) and gliocytes increased. The results showed statistical difference comparing with control group (P &lt; 0.05)., Conclusion: The brainstem encephalitis can be used to diagnose fatal EV71 infection in infants. EV71 can invade the brainstem via hematogenous route. ICAM-1 may play an important role in the pathogenic process.

Atropine-induced heart damage is associated with changes in the expression of various enzymes and proteins. The purpose of this study was to investigate atropine-induced alterations in cardiac E-cadherin and 5-hydroxytryptamine (5-HT) after atropine administration. Male Wistar rats were randomly divided into two groups: a control group and an atropine group. The atropine group intraperitoneally received atropine at a single dose of 15 mg/kg for 7 days; the controls received the same amount of saline via the same route. On Day 8, the rats were anesthetized, and a thoracotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of E-cadherin and 5-HT. Sections were analyzed by digital image analysis. Cardiac protein expression of E-cadherin and 5-HT was altered after atropine‑induced toxicity in the rat. The expression levels of E-cadherin and 5-HT were significantly decreased after atropine treatment, supported by IOD analysis, when compared with the control (P<0.05). The current findings indicate that such changes would be reflected in abnormal cardiac function, and these proteins may be useful for revealing the mechanisms underlying atropine-induced toxicity and may also provide various clues for further research.

Heart damage induced by chlorpromazine (CPZ) toxicity is associated with changes in the expression of various enzymes and proteins. This study aimed to investigate CPZ‑induced alterations in cardiac E-cadherin and caveolin-1 (cav-1) after CPZ administration. Male Wistar rats were randomly divided into two groups: a control group and a CPZ group. The CPZ group was administered CPZ intraperitoneally at a single dose of 10 mg/kg for 21 days; the controls were given the same amount of saline via the same route. On Day 22, the rats were anesthetized, and a thoracotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of E-cadherin and cav-1. Sections were analyzed by digital image analysis. Results of the present study revealed that cardiac protein expression of E-cadherin and cav-1 was altered after CPZ-induced toxicity in the rat. The expression of E-cadherin was significantly reduced, while expression of cav-1 was significantly increased after CPZ treatment, as supported by integrated optical density analysis, compared with the control (P<0.05). The current findings indicate that such changes in the expression of E-cadherin and cav-1 may be reflected in abnormal cardiac function, and these proteins may be useful in revealing the mechanisms underlying CPZ-induced toxicity and may also provide additional insight for further research.

Gentamicin (GM)-induced heart damage is associated with alterations in expression levels of various enzymes and proteins. The aim of the present study was to investigate GM-induced alterations in cardiac α-enolase and caveolin after GM administration. Male Wistar rats were randomly divided into two groups: a control group and a GM group. The GM group intraperitoneally received GM at a single dose of 7 mg/kg for 8 days, while the controls were given the same amount of saline via the same route. On Day 9, the rats were anesthetized and a thoracotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of α-enolase and caveolin. Sections were analyzed by digital image analysis. Our results revealed that cardiac protein expression of α-enolase and caveolin was altered after GM-induced toxicity in the rat. The expression of α-enolase and caveolin was significantly increased after GM-induced toxicity, as determined by integrated optical density analysis, when compared with the control (P<0.05). The current findings indicate that such changes in protein expression may be reflected in abnormal cardiac function, and the proteins identified in this study may be useful for elucidating the mechanisms underlying GM-induced toxicity and may also provide various clues for further investigations.

Cisplatin-induced heart damage is associated with enzymes and protein alterations. The purpose of this study was to investigate cisplatin-induced alterations in cardiac endothelin (ET)-1 and type III collagen following administration of cisplatin. Male Wistar rats were randomly divided into two groups: a control and a cisplatin group. The cisplatin group received cisplatin intraperitoneally (i.p.) at a single dose of 7 mg/kg on day 6, while the controls were given the same amount of saline via the same route. On day 11, the rats were anesthetized and a thoracotomy was performed on all animals. Immunohistochemical analysis was performed to evaluate the protein expression of ET-1 and type III collagen. Sections were analyzed by digital image analysis. Results showed that the cardiac protein expression of ET-1 and type III collagen was altered following cisplatin-induced toxicity in rats. The expression of ET-1 and type III collagen was significantly increased after cisplatin treatment, supported by integrated optical density, when compared to the control group (P<0.05). The present findings indicate that such alterations may be reflected in abnormal cardiac function. Additionally, the proteins identified in this study may benefit investigations into the mechanisms underlying cisplatin-induced toxicity, thereby providing the necessary evidence for further research.

Objective: To examine the role of domain II of hepatitis C virus (HCV) 5' noncoding region (5' NCR) in its translation initiation activity., Methods: The fragment of HCV 5' NCR with deletions of 5'-proximal 118 nucleotides were amplified with PCR, then was used to substitute for the full length HCV 5' NCR of plasmid pCMVNCRluc, a firefly luciferase (Fluc) eukaryotic expression plasmid regulated by HCV 5' NCR, to generate recombinant plasmid pCNl-d3. pCMVNCRluc, pCNl-d2 (modified pCMVNCRluc with deletions of HCV 5' NCR ntl-43) and pCNl-d3 were transfected into HepG2 cells with liposome transfection protocol, respectively, and the relative luciferase activity was measured to analyze the regulatory effect of the truncated 5'NCR on Flue gene expression. Meanwhile the Flue mRNA levels were detected by RT-PCR., Results: The recombinant plasmid was successfully constructed. The Flue mRNA levels of the 3 plasmids were not significantly different (P > 0.05), and there were also no significant difference between the relative luciferase activity of plasmids pCNl-d2 and pCMVNCRluc (P > 0.05). However, that of pCNl-d3 was decreased significantly (P < 0.01, compared with pCNl-d2 or pCMVNCRluc)., Conclusion: Structural domain II of HCV 5' NCR plays an important role in its translation initiation activity.

Objective: To investigate the effects of serum from crush injury rats on vascular endothelial cell apoptosis and their potential mechanism., Methods: Bovine aorta endothelial cells were cultured in vitro and the effects of serum from crush injury rats on cell apoptosis and intracellular calcium concentration ([Ca2+]i) were observed. Meanwhile, the levels of rat blood plasma endothelin-1 (ET-1) and atrial natriuretic peptide(ANP) were measured., Results: Compared with normal rat serum treatment, the cell apoptosis rate decreased from (8.26+/-1.75)% to (2.75+/-0.90)%, while the concentration of [Ca2+]i increased from (96.98+/-3.95) to (118.79+/-3.22) nmol/L in serum from crush injury rats, respectively. The concentration of ET-1 and ANP increased significantly in crush injury rat serum., Conclusion: Serum from crush injury rats could inhibit apoptosis of the vascular endothelial cells. These effects may be related to increased level of [Ca2+]i mediated by ET-1 and ANP.

Objective: To study the influence of human cytomegalovirus (HCMV) infection on cell cycle and the expression of replication licensing factor Cdt1 in human embryonic lung fibroblastic (HEL) cells and to explore the pathogenesis of HCMV infection., Methods: HEL cells were synchronized in the G0/G1 phase by the serum starvation method. The synchronized HEL cells were infected with HCMV, and those that were not subjected to HCMV infection were used as the control group. The HEL cells were harvested at 12, 24, 48, 72 and 96 hrs of HCMV infection. The cell cycle of HEL cells was detected by the flow cytometry. The expression of Cdt1 mRNA in HEL cells was determined by reverse transcription-polymerase chain reaction (RT-PCR)., Results: The cells in the G1 phase in the control group was significantly more than in the HCMV-infected group 12 and 24 hrs after infection (P < 0.01). The expression of Cdt1 mRNA in the HCMV-infected group was significantly lower 12 and 24 hrs after infection but increased significantly 48 hrs after infection compared with the control group (P < 0.05). The expression of Cdt1 mRNA reached a peak at 12 hrs of infection in the control group, but at 48 hrs of infection in the HCMV-infected group, which markedly lagged behind the control group., Conclusions: HCMV infection arrests the cell cycle of HEL cells at the G1 phase. HCMV infection makes Cdt1 expression delay. HCMV infection can interfere cell cycle of HEL cells possibly through affecting the expression of Cdt1.

Objective: To observe the effects of heroin on intracellular free Ca2+ in rat myocardium., Methods: The effects of heroin on intracellular free Ca2+ were observed in cultured neonatal rat myocardium by measuring intracellular free Ca2+ concentration using calcium fluorescent probe Flou-3/AM and laser scanning confocal microscope., Results: Different doses and concentrations of heroin appeared to have different effects on intracellular free Ca2+ concentrations, with a dosage dependent short linear increase in the fluorescence intensity (i.e., Ca2+ concentration) leading to [Ca2+]i peak., Conclusion: Heroin could affect concentrations of [Ca2+]i in myocardium and its dosage related effect needs further investigation.

Objective: To investigate whether heroin can directly induce apoptosis in primary cultured cortical neurons of rat's brain., Methods: Cultured primary neurons cultures were obtained from cerebral cortex of embryo rats. After 7 days, the cells were incubated with different concentrations of heroin (purity-80%) for 24 hours. The neuronal survival was assessed by cell viability counting with fluorescent diacetate (FDA) staining. The morphological and biochemical changes were observed with Hoechst 33258 fluorescent staining and then analyzed by agarose gel electrophoresis, respectively., Results: After treatment with different concentrations of heroin, the neurons showed a decreased survival rate in a dose dependent manner, and there was a significant difference in the survival rate between the heroin group and the control group (P < 0.05). When exposed to different concentrations of heroin, neurons exhibited the morphological and biochemical features of apoptosis, including cell shrinkage, neurite degeneration, network disappearance, condensation and aggregation of nuclear chromatin, and the formation of DNA ladders. With the increase of heroin concentration of rat's brain more apoptotic bodies were seen., Conclusion: Heroin can directly induce apoptosis in primary cultured cortical neurons in rat's brain.