Objective To investigate the role of transcription factor EB(TFEB)in endotoxin-tolerant macrophages.Methods The RAW264.7 cells were divided into blank group(DMEM medium),LPS 5 group(5 ng/mL LPS treatment for 4 h),LPS 100 group(100 ng/mL LPS treatment for 4 h),and tolerance group(5 ng/mL LPS for 12 h followed by 100 ng/mL LPS for 4 h).The releases of inflammatory factors TNF-α and IL-6 were measured using ELISA.Western blotting and immunofluorescence assay were used to evaluate the distribution of autophagy-related proteins LC3 and P62,as well as TFEB in the cytoplasm and nucleus.Lentiviral overexpression of TFEB or siRNA-mediated knockdown of TFEB were performed to observe the changes in autophagy levels and bacterial clearance ability in the tolerant cells.Results The cells in the tolerance group had significantly lower contents of TNF-α and IL-6,as well as reduced bacterial clearance ability(P<0.01),down-regulated LC3 expression while up-regulated P62 level,and decreased expression of TFEB in both the cytoplasm and nucleus(P<0.01)when compared with the cells of the LPS 100 group.Overexpression of TFEB significantly increased LC3 level,reduced P62 level,and enhanced bacterial clearance ability in the endotoxin-tolerant cells(P<0.01).In contrast,siRNA-mediated knockdown of TFEB had no significant impacts on LC3 and P62 expression levels or bacterial clearance ability.Conclusion Overexpression of TFEB can restore the autophagy of endotoxin-tolerant cells and enhance their bacterial clearance capacity,thereby alleviating the immunosuppressive state of sepsis.These findings suggest that TFEB holds promise as a potential therapeutic target for the prevention and treatment of sepsis.

Objective To investigate the role and underlying mechanism of activating transcription factor 3(ATF3)in suppressing lipopolysaccharide(LPS)-induced autophagy and inflammatory responses in macrophages.Methods Firstly,the gene expression omnibus(GEO)database was used to analyze ATF3 expression in peripheral blood mononuclear cells(PBMCs)from sepsis patients,and gene set enrichment analysis(GSEA)was performed to identify enriched signaling pathways.Secondly,RAW264.7 macrophages were divided into a blank control group and an LPS-stimulated group(100 ng/mL LPS).Western blotting and immunofluorescence assay were used to detect ATF3 protein expression and observe its subcellular localization,respectively.Lentiviral transduction was used to generate ATF3 knockdown and overexpression cell lines to evaluate their effects on cytokine release and bacterial clearance.Cleavage Under Targets and Tagmentation(CUT&Tag)sequencing was employed to identify downstream target genes transcriptionally regulated by ATF3.Furthermore,the impact of ATF3 knockdown or overexpression on autophagy-related gene 5(ATG5),autophagy-related gene 16-like 1(ATG16L1),and autophagy levels was evaluated.Results GEO analysis revealed that ATF3 expression was significantly elevated in PBMCs from sepsis patients(P<0.01),and GSEA showed significant enrichment of autophagy-related and inflammation-related pathways(P<0.01).In RAW264.7 cells,100 ng/mL LPS stimulation significantly increased ATF3 expression in the nucleus than the blank control group(P<0.01).ATF3 knockdown led to increased secretions of TNF-α and IL-6 and enhanced bacterial clearance of macrophages(P<0.01),whereas ATF3 overexpression significantly suppressed TNF-α and IL-6 releases,and remained bacterial clearance at a low level when compared with the conditions in the negative control(NC)group(P<0.01).CUT&Tag results demonstrated that ATF3 was enriched at the promoter regions of key autophagy genes Atg5 and Atg16l1.Compared with the NC group,ATF3 knockdown significantly up-regulated the protein levels of LC3-II/I,ATG5,and ATG16L1 while decreased p62 expression(P<0.01).Conversely,ATF3 overexpression inhibited the expression of LC3-II/I,ATG5,and ATG16L1(P<0.01),but had no significant effect on p62 level.Conclusion Sepsis induces elevated ATF3 expression in macrophages,and suppresses autophagic activity and down-regulates pro-inflammatory cytokines TNF-α and IL-6,which probably mediated by ATF3 regulating transcription of ATG5 and ATG16L1,suggesting ATF3 as a potential therapeutic target for autophagy-inflammation imbalance.

Objective:To investigate the involvement of ferroptosis mediated by glutathione peroxidase 4(Gpx4)in acute lung injury in rats with sepsis,as well as the intervention mechanism of curcumin(Cur).Methods:Twenty-four rats were randomly divided into three groups,with 8 rats in each group:Sham group,Sepsis group and Cur group.The rat model of acute lung injury in sepsis was established by cecal ligation and puncture(CLP).The Sham group only underwent laparotomy and closure.Both Sepsis group and Cur group underwent CLP,Cur group was injected with curcumin(200 mg/kg)intraperitoneally 1 hour after modeling,and the administra-tion was repeated 24 hours later.The lung tissues of the rats were sampled 48 hours after surgery,and the wet/dry weight of lung tissue ratio(W/D)of lung tissues in each group was measured.Morphological changes of lung tissues were observed by HE staining.The con-tents of GSH,MDA,and Fe2+and the levels of inflammatory factors including TNF-α,IL-6,and IL-1β in the lung tissues were mea-sured using test kits.Western blot was used to detect the expressions of nuclear factor E2-related factor 2(Nrf2)and the key regulatory protein of ferroptosis Gpx4.Apoptosis of alveolar epithelial cells was detected by TUNEL method.Ultrastructural changes of alveolar epithelial cells were observed by transmission electron microscopy.Results:Compared with the Sham group,lung tissues in the Sepsis group showed an increased W/D(P<0.05),significantly higher levels of inflammatory factors TNF-α,IL-6,and IL-1β and contents of MDA and Fe2+(P<0.05),decreased content of GSH,up-regulated expression of Nrf2,and down-regulated expression of Gpx4(P<0.05),the type Ⅱ alveolar epithelial cells were seen to be edematous,congested,and infiltrated with inflammatory cells via light microscopy,and ferroptosis signs such as mitochondrial crinkling,thickened bilayer membrane density,and reduced or broken cristae were seen via transmission electron microscopy.Compared with Sepsis group,lung tissues in the Cur group showed decreased W/D(P<0.05),lower levels of TNF-α,IL-6 and IL-1β,lower contents of MDA and Fe2+,increased content of GSH,down-regulated expression of Nrf2,and up-regulated expression of Gpx4(P<0.05),the pathological changes of lung tissues were significantly reduced.Conclusion:Curcumin reduces sepsis-induced acute lung injury in rats,and the mechanism is related to inhibiting inflamma-tory response and GPX4-mediated ferroptosis.

Objectives:This study aimed to analyze the clinical and molecular characteristics of carbapenem-resistant Enterobacteriaceae (CRE) bloodstream infection (BSI) in patients with hematological diseases and to explore prognostic risk factors.Methods:This retrospective study included patients with hematologic diseases with CRE BSI at the Institute of Hematology and Blood Diseases Hospital from January 2015 to December 2022. The clinical features, carbapenemase test results, antimicrobial treatments, and outcomes were analyzed.Results:A total of 120 patients developed CRE BSI. Escherichia coli (58/120, 48.3%) was the most prevalent Enterobacteriaceae, followed by Klebsiella pneumoniae (52/120, 43.3%). A total of 93 CRE strains were tested for carbapenemase, of which 75 strains produced carbapenemase (metalloenzyme: 51 strains; serine enzyme: 24 strains). The 30-day mortality rate after BSI was 24.2% (29/120). Univariate analysis revealed significantly lower mortality in patients treated with the ceftazidime-avibactam-containing regimen than in those treated with other antibiotics (7.8% vs 36.2%, P<0.001). Moreover, initiating active therapy within 24 h of BSI onset significantly reduced mortality (15.0% vs 33.3%, P=0.019). The proportion of patients with CRE colonization receiving active therapy within 12 and 24 h was significantly higher compared with patients without colonization (12 h: 14.5% vs 34.1%, P=0.012; 24 h: 40.8% vs 65.9%, P=0.008). Multivariate analysis revealed that septic shock ( HR=24.436, 95% CI 4.148 - 143.966, P<0.001) and pulmonary infection ( HR=9.346, 95% CI 2.718-32.140, P<0.001) were independent risk factors for death within 30 days. Appropriate therapy was initiated within 24 h ( HR=0.225, 95% CI 0.059 - 0.851, P=0.028), and treatment with the ceftazidime-avibactam-containing regimen ( HR=0.082, 95% CI 0.018-0.362, P=0.001) significantly reduced mortality. Conclusion:The prognosis of CRE BSI in patients with hematological diseases is poor. Timely, appropriate therapy and receipt of a ceftazidime-avibactam-containing regimen can improve survival and prognosis.

Objectives:This study aimed to analyze the clinical and molecular characteristics of carbapenem-resistant Enterobacteriaceae (CRE) bloodstream infection (BSI) in patients with hematological diseases and to explore prognostic risk factors.Methods:This retrospective study included patients with hematologic diseases with CRE BSI at the Institute of Hematology and Blood Diseases Hospital from January 2015 to December 2022. The clinical features, carbapenemase test results, antimicrobial treatments, and outcomes were analyzed.Results:A total of 120 patients developed CRE BSI. Escherichia coli (58/120, 48.3%) was the most prevalent Enterobacteriaceae, followed by Klebsiella pneumoniae (52/120, 43.3%). A total of 93 CRE strains were tested for carbapenemase, of which 75 strains produced carbapenemase (metalloenzyme: 51 strains; serine enzyme: 24 strains). The 30-day mortality rate after BSI was 24.2% (29/120). Univariate analysis revealed significantly lower mortality in patients treated with the ceftazidime-avibactam-containing regimen than in those treated with other antibiotics (7.8% vs 36.2%, P<0.001). Moreover, initiating active therapy within 24 h of BSI onset significantly reduced mortality (15.0% vs 33.3%, P=0.019). The proportion of patients with CRE colonization receiving active therapy within 12 and 24 h was significantly higher compared with patients without colonization (12 h: 14.5% vs 34.1%, P=0.012; 24 h: 40.8% vs 65.9%, P=0.008). Multivariate analysis revealed that septic shock ( HR=24.436, 95% CI 4.148 - 143.966, P<0.001) and pulmonary infection ( HR=9.346, 95% CI 2.718-32.140, P<0.001) were independent risk factors for death within 30 days. Appropriate therapy was initiated within 24 h ( HR=0.225, 95% CI 0.059 - 0.851, P=0.028), and treatment with the ceftazidime-avibactam-containing regimen ( HR=0.082, 95% CI 0.018-0.362, P=0.001) significantly reduced mortality. Conclusion:The prognosis of CRE BSI in patients with hematological diseases is poor. Timely, appropriate therapy and receipt of a ceftazidime-avibactam-containing regimen can improve survival and prognosis.

Telephone follow-up is one of the important ways to follow up patients. High-quality follow-up can benefit both doctors and patients. However, clinical research-related follow-up is often faced with problems such as time-consuming, laborious and poor patient compliance. The authors belong to a team that has been committed to the study of patient-reported outcomes for a long time. The team has carried out long-term follow-up of symptoms, daily function and postoperative complications of more than 1 000 patients after lung cancer surgery, and accumulated certain experience. In this paper, the experience of telephone follow-up was summarized and discussed with relevant literatures from the aspects of clarifying the purpose of clinical research follow-up, understanding the needs of patients in follow-up, and using follow-up skills.

【Objective】 To investigate the clinical value of transrectal contrast-enhanced ultrasound (CEUS) in the diagnosis of prostate cancer in different total prostate specific antigen (tPSA) intervals. 【Methods】 According to serum tPSA levels, 96 patients meeting the inclusion criteria were divided into 3 groups:4-10 ng/mL, >10-20 ng/mL and >20 ng/mL groups. All patients underwent transrectal CEUS. With pathological results as reference, the diagnostic value of transrectal CEUS in different tPSA intervals was evaluated. 【Results】 Of the 96 cases, 62 were confirmed by pathology as prostate cancer and 34 as benign prostatic hyperplasia (BPH). The main perfusion characteristics of prostate cancer under CEUS were rapid enhancement (64.52%), rapid clearance (70.97%), uneven enhancement (83.87%) and high enhancement (61.29%);the main characteristics of BPH were non-rapid enhancement (70.59%), non-rapid clearance (73.53%), uniform enhancement (76.47%) and non-high enhancement (52.94%). There were significant differences in terms of enhancement speed, clearance speed and enhancement uniformity between prostate cancer and BPH (P<0.05), but no significant difference in the enhancement intensity. The sensitivity of transrectal CEUS in the diagnosis of prostate cancer in low, medium and high tPSA groups were 58.33%, 70.37% and 95.65%, the specificity were 83.33%, 76.92% and 66.67%, and the accuracy were 73.33%, 72.50% and 92.31%, respectively. Transrectal CEUS showed consistency at different serum tPSA levels for the diagnosis of prostate cancer, with statistical significance. Moreover, in the 4.0 ng/mL ≤tPSA<10.0 ng/mL group, the diagnostic specificity was the highest. 【Conclusion】 Transrectal CEUS is helpful in the differential diagnosis of benign and malignant prostatic lesions, especially for patients with different serum tPSA levels.

Intraoperative ultrasound assisted localization is routinely used in the surgical treatment of completely endogenous renal cell tumor, however the localization and guidance ability of conventional ultrasound will decline to a certain extent for isoechoic renal tumor. A 62 years old female patient with right renal tumor was reported. The tumor diameter was about 2.3 cm× 1.7 cm, equivalent to the isoechoic of kidney. Laparoscopic partial nephrectomy was performed under the real-time guidance of contrast-enhanced ultrasound. The tumor was found to be lack of blood supply during the operation, and the tumor contour was clearly developed by contrast agent.The operation was successfully completed, and the pathological diagnosis was polycystic renal tumor with low malignant potential.The incisional margin was negative.The patient recovered well after operation without complications.No recurrence or metastasis was found after 6 months of follow-up.The renal function was good.

Background Prolonged awkward postures during occupational activities can lead to excessive musculoskeletal load on the wrist of workers and symptoms such as wrist pain or discomfort. Objective To survey the prevalence of wrist pain among workers in 10 key industries and analyze its correlation with wrist working postures. Methods By using stratified cluster sampling method, workers from 10 key industries, such as footwear manufacturing industry, shipbuilding manufacturing industry, and automobile manufacturing industry, were selected from seven regions in North China, East China, Central China, South China, Southwest China, Northwest China, and Northeast China. The demographic information, wrist working postures, pain in wrist of the workers were collected through a cross-sectional survey. Pearson χ² test was used to compare prevalence by selected factors, trend χ² test for between group comparison, and unconditional logistic regression models for the association of wrist working postures with wrist pain. Results There were 64052 workers enrolled in this survey, and 56286 provided valid questionnaires (the effective rate was 87.8%). According to the survey, the prevalence of wrist pain was 23.3% (13112/56286), and the industries with higher prevalences were footwear manufacturing (27.1%, 1927/7106), automobile manufacturing (24.9%, 5378/21560), and shipbuilding and related equipment manufacturing (24.4%, 850/3488) industries. Finger pinching (OR=2.09, 95%CI: 1.95-2.24), frequent wrist bending (OR=2.03, 95%CI: 1.92-2.15), fixed wrist bending (OR=1.77, 95%CI: 1.69-1.85), wrist on hard edge (OR=1.34, 95%CI: 1.28-1.40), and arms over shoulders (OR=1.11, 95%CI: 1.05-1.17) increased the risk of reporting wrist pain. Conclusion Awkward postures are related to wrist pain among workers in selected 10 key industries. The related factors are wrist on hard edge, frequent wrist bending, finger pinching, fixed wrist bending, and arms over shoulders.

Objective:Based on the regulatory effect of curcumin (Cur) on inflammation and iron death, to explore the mechanism of Cur protecting against cerebral ischemia-reperfusion injury (CIRI).Methods:A rat model of middle cerebral artery occlusion (MCAO) was established by the modified suture-occluded method. The modeled SD rats were randomly divided into the Sham group, CIRI group and Cur group. The neurobehavioral score of rats was measured by the Longa method. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in the brain tissue of rats in each group. Furthermore, the contents of glutathione (GSH), malondialdehyde (MDA) and Fe 2+, as well as the levels of the inflammatory factors tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 in the ischemic cerebral cortex, were detected by corresponding testing kits. Western blotting was applied to detect the expression of glutathione peroxidase 4 (GPX4), a key regulatory protein of ferroptosis in the cerebral cortex. In addition, neuronal apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, and ultrastructural changes in neurons in the cerebral cortex were observed under a transmission electron microscope. Results:Compared with the CIRI group, the Cur group showed decreased neurobehavioral scores, significantly reduced contents of MDA, Fe 2+, TNF-α, IL-1β and IL-6 (all P<0.05), but obviously increased content of GSH and protein expression of GPX4 (both P<0.05). Further pathological examination revealed edema, rupture and necrosis of neurons in the CIRI group, while mild edema and a small number of necrotic cells were observed in the Cur group only. The results of TUNEL staining indicated that the rate of neuronal apoptosis in the Cur group was lower than that in the CIRI group, with a statistically significant difference between groups [(23.6±3.5)% vs. (36.8±4.2)%; P<0.05]. In addition, under the transmission electron microscope, the CIRI group had a reduced volume of mitochondria, thickened double-layer membrane structure, and decreased or disappeared mitochondrial cristae, while the Cur group showed partial margination of nuclear chromatin and alleviated damage to mitochondria. Conclusions:Cur could attenuate CIRI, and its neuroprotective mechanism may be related to the inhibition of the inflammatory response and GPX4-mediated ferroptosis.