Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathway.

As a pattern recognition receptor, Toll-like receptor 4 (TLR4) is crucial for the development and progression of acute kidney injury (AKI). This study aims to explore whether the deubiquitinase Usp9x influences the TLR4/NF-B pathway to cause sepsis-induced acute kidney injury (S-AKI). The model of AKI was established in Sprague-Dawley rats using the cecal ligation and puncture (CLP) method, while renal tubular epithelial cell NRK-52E was stimulated with lipopolysaccharide (LPS) in vitro. All plasmids were transfected into NRK-52E cells according to the indicated group. The deubiquitinase of TLR4 was predicted by the online prediction software Ubibrowser. Subsequently, Western blot and Pearson correlation analysis identified Usp9x protein as a potential candidate. Co-IP analysis verified the interaction between TLR4 and Usp9x. Further research revealed that overexpression of Usp9x inhibited degradation of TLR4 protein by downregulating its ubiquitination modification levels. Both in vivo and in vitro experiments observed that interference with Usp9x effectively alleviated the inflammatory response and apoptosis of renal tubular epithelial cells (RTECs) induced by CLP or LPS, whereas overexpression of TLR4 reversed this situation. Transfection with sh-Usp9x in NRK-52E cells suppressed the expression of proteins associated with the TLR4/NF-κB pathway induced by LPS. Moreover, the overexpression of TLR4 reversed the effect of sh-Usp9x transfection. Therefore, the deubiquitinase Usp9x interacts with TLR4, leading to the upregulation of its expression through deubiquitination modification, and the activation of the TLR4/NF-κB signaling pathway, thereby promoting inflammation and apoptosis in renal tubular epithelial cells and contributing to sepsis-induced acute kidney injury.

An investigation on cervical cancer and human papillomavirus vaccine knowledge, and analysis of influencing factors for choosing domestic or imported 2vHPV vaccine among females in Shenzhen, China.

In 2020, the domestic (Chinese native) 2v human papillomavirus (HPV) vaccine was approved for use in females. At present, there are obvious differences in demand for different HPV vaccines. We aimed to investigate knowledge level of cervical cancer and HPV vaccine and its influencing factors among the eligible female recipients in Shenzhen, China, and to analyze the factors influencing choice of 2vHPV vaccine (domestic or imported) would be selected. A self-administered questionnaire was carried out on this investigation, and respondents were selected by random sampling method conducted by vaccination doctors. A total of 1197 valid questionnaires were collected, of which 729 (60.9%) were vaccinated with domestic vaccines and 468 (39.1%) were vaccinated with imported. Four hundred and fifty (61.7%) and 306 (65.4%) got a passing grade, respectively (χ2 = 1.637, P = .201). Logistic regression analysis showed that age (P = .002), ethnicity (P < .001), duration of residence in Shenzhen (P < .001), educational level (P < .001) and occupation (P < .001) were significant. It also showed that the manufacturers (P < .001), efficacy (P < .001), safety (P < .001), cognitive approach (P < .001), public opinion (P < .001), convenient acquisition (P < .001) and knowledge reserve (P = .035) were statistically significant. While price (P = .371) and doctor's suggestion (P = .114) were not. In conclusion, eligible female recipients had a high degree of knowledge regarding cervical cancer and HPV vaccine, education level and occupation were the most important factors for scores. Domestic 2vHPV vaccine was more widely utilized than imported, manufacturer, efficacy, safety, cognitive approach, public opinion, convenient acquisition, and knowledge reserve had an impact on selection for recipients, while price and doctor's suggestion did not.

Circular RNA circ_0003420 mediates inflammation in sepsis-induced liver damage by downregulating neuronal PAS domain protein 4.

OBJECTIVE: Our aim was to investigate whether circular RNA (circRNA) circ_0003420 mediates inflammation in sepsis-induced liver damage and to determine the mechanism involved. MATERIALS AND METHODS: Liver tissue samples from patients with sepsis and healthy subjects were used to identify differentially expressed circRNAs. Additionally, Kupffer cells were treated with lipopolysaccharide (LPS) to establish an in vitro model of sepsis-induced liver damage. Cell viability and proliferation were measured with a cell counting kit-8 and 5-ethynyl-2'-deoxyuridine (EdU) labeling, respectively. Relative mRNA and protein levels of IL-6, IL-1ß, tumor necrosis factor (TNF)-α, and neuronal PAS domain protein 4 (NPAS4) were determined via reverse-transcription quantitative PCR and western blotting, respectively. RESULTS: We observed circ_0003420 upregulation accompanied by NPAS4 downregulation in liver samples from patients with sepsis-associated damage and in Kupffer cells treated with LPS. Results of in vitro experiments indicated that LPS treatment reduced cell viability and induced well-pronounced apoptosis and inflammatory signs. Circ_0003420 silencing counteracted LPS's influence on cell proliferation, apoptosis, and inflammation signs. Bioinformatics and a dual-luciferase reporter assay revealed that circ_0003420 targets NPAS4 mRNA and negatively correlates with NPAS4 expression. Moreover, NPAS4 knockdown recovered the apoptosis rate and expression levels of inflammatory cytokines in the LPS-treated circ_0003420 knockdown cells, whereas NPAS4 overexpression had similar effects on Kupffer cell properties as circ_0003420 silencing. CONCLUSION: We demonstrate that circ_0003420 targets NPAS4 mRNA thereby mediating the cell damage and inflammation caused by LPS. This study provides a possible target for treatment of liver damage induced by sepsis.

"Awake" Extracorporeal Membrane Oxygenation Combined With Continuous Renal Replacement Therapy For the Treatment of Severe Chemical Gas Inhalation Lung Injury.

Lung injury caused by chemical gas inhalation is a common clinically severe disease that very easily progresses to acute respiratory distress syndrome (ARDS). Traditional respiratory support consists mainly of mechanical ventilation, but the prognosis of this condition is still poor. "Awake" extracorporeal membrane oxygenation (ECMO) maintains oxygenation, improves ventilation, adequately allows the injured lungs to rest, and avoids complications associated with sedation, intubation, and mechanical ventilation. Continuous renal replacement therapy (CRRT) can provide better fluid management and reduce pulmonary edema. Herein, we describe the case of a patient with severe chemical gas inhalation lung injury who failed to respond to traditional mechanical ventilation and was subsequently treated with awake ECMO combined with CRRT.

TSLP induces a proinflammatory phenotype in circulating innate cells and predicts prognosis in sepsis patients.

Thymic stromal lymphopoietin (TSLP) has been identified as a crucial inflammatory cytokine in immune homeostasis. Previous studies have reported conflicting effects of TSLP on sepsis in mice, and the effect of TSLP on sepsis in humans has not been investigated. In this study, we used the ELISA to measure serum levels of TSLP in patients with sepsis, and used flow cytometry and ELISA to evaluate the proinflammatory phenotype of circulating immune cells. In addition, we used quantitative RT-PCR to examine the expression of proinflammatory cytokines [interleukin (IL)-1ß, IL-6, tumor necrosis factor-α, transferrin growth factor-ß, IL-10, and matrix metalloproteinase] between patients with high and low levels of TSLP. Flow cytometry analysis was performed to evaluate the phagocytic and respiratory burst of circulating neutrophils. A significant increase in the production of proinflammatory cytokines by nonclassical monocytes and the number of interferon (IFN)-γ+  CD4+ monocytes was observed in patients with high levels of TSLP. Furthermore, the number of IL-10+ regulatory T cells was observed to be increased in patients with high levels of TSLP. We found that TSLP values greater than 350 pg·mL-1 were associated with a higher mortality rate and longer stays in intensive care (sensitivity of 89% and specificity of 79%). In patients with low levels of neutrophils, the area under curve was only 0.71 (based on the cutoff value in the diagnostic test evaluation; sensitivity of 62% and specificity of 68%). Our findings suggest that the serum levels of TSLP may be suitable as a biomarker for prediction of prognosis in a subgroup of patients with sepsis who are exhibiting hyperleukocytosis and a high neutrophil ratio.

[Effect of angiotensin II on the expression of aquaporin 1 in lung of rats following acute lung injury].

OBJECTIVE: To investigate the effect of angiotensin II (Ang II) on the expression of aquaporin 1 (AQP1) in lung of rats with acute lung injury (ALI) and the role of Ang II in the formation of lung edema. METHODS: Forty healthy Sprague-Dawley (SD) rats were randomly divided into sham-operated group, model group, Ang II receptor blocker pretreatment group, and Ang II receptor blocker treatment group according to random digits table, with 10 rats in each group. ALI model of rats was reproduced with administration of endotoxin after hemorrhagic shock. In rats of pretreatment group Ang II receptor blocker 30 microg/kg was given 30 minutes before lipopolysaccharide (LPS) injection; rats in treatment group Ang II receptor blocker 30 microg/kg was given 30 minutes after LPS injection; rats in model group received 30 microg/kg normal saline 30 minutes before and after LPS injection. Rats were sacrificed 6 hours after model establishment, samples of venous blood and lung tissue were collected, radioimmunoassay was used to measure the level of tumor necrosis factor-alpha (TNF-alpha) in serum and the expression of Ang II in lung tissue, ratio of wet to dry (W/D) weight of lung tissue was calculated, reverse transcription-polymerase chain reaction was used to measure the expression of AQP1 mRNA in lung tissue. RESULTS: Compared with rats of sham-operated group, the level of TNF-alpha in venous blood, W/D ratio and the expression of Ang II in lung tissue increased significantly, the expression of AQP1 mRNA in lung tissue decreased significantly in rats of ALI. Compared with rats of model group, the level of TNF-alpha in venous blood (microg/L) decreased significantly (4.79+/-0.24, 5.55+/-0.36 vs. 6.34+/-0.31, both P<0.05), W/D ratio decreased significantly (4.34+/-0.23, 4.85+/-0.20 vs. 5.41+/-0.26, both P<0.05), the expression of AQP1 mRNA in lung tissue increased significantly (0.854+/-0.067, 0.727+/-0.081 vs. 0.358+/-0.071, both P<0.05), and the expression of Ang II in lung tissue (ng/g) decreased to some extent (172.19+/-15.82, 202.82+/-20.47 vs. 245.88+/-26.31), but without statistical significance (both P>0.05) in rats of pretreatment group and treatment group. The expression of AQP1 mRNA was negatively correlated with both the level of Ang II and W/D ratio (r1=-0.782, r2=-0.726, both P<0.05). CONCLUSION: During ALI, Ang II may downregulate the expression of AQP1 mRNA in lung tissue directly or through inflammatory mediators, Ang II may play a role in the formation of lung edema.