Inflammation-dependent activation of NCOA2 associates with p300 and c-MYC/Max heterodimer to transactivate RUNX2-AS1 and mediate RUNX2 downstream bone differentiation genes in the pathology of septic nonunion.

Septic nonunion (SN) is a common bone disorder caused by the failure of fracture healing. Local inflammation in fracture sites often causes SN; however, little is known about the molecular mechanisms of SN pathology. Herein, we identified a significant upregulation of the long non-coding RNA (lncRNA) RUNX2-AS1 (Runt-related Transcription Factor 2-Antisense 1) in the biopsies of SN patients. Overexpression or knockdown of RUNX2-AS1 in vitro could inhibit or induce, respectively, the expression of RUNX2 and RUNX2-downstream target genes, including ALPL (Alkaline Phosphatase), COL1A1 (Collagen Type I Alpha 1 Chain), IBSP (Integrin Binding Sialoprotein), MMP13 (Matrix Metallopeptidases), and SPP1 (Secreted Phosphoprotein 1), which are involved in bone differentiation. Mechanically, we demonstrated that a transcription factor c-MYC could assemble a transcriptional complex with its partner Max, a histone acetyltransferase p300, and nuclear receptor coactivator 2 (NCOA2), and this complex then bound to the promoter of RUNX2-AS1 to transactivate its expression. The mRNA and protein levels of NCOA2 were dose-dependently increased by treatment with lipopolysaccharide(LPS), a well-known inflammation trigger. LPS exposure increased the enrichment of the NCOA2-p300-c-MYC/Max complex on the RUNX2-AS1 promoter to activate its expression, thereby downregulating the expression of RUNX2 and RUNX2-downstream target genes. Depletion of NCOA2 reversed the expression of RUNX2-AS1, RUNX2, and RUNX2 target genes following LPS exposure. Taken together, our results demonstrate a new signaling pathway that contributes to the pathology of SN and may aid in preventing SN progression.

Enhancement of autophagy flux by isopsoralen ameliorates interleukin-1ß-stimulated apoptosis in rat chondrocytes.

Chondrocyte apoptosis contributes to the pathogenesis of cartilage degeneration in osteoarthritis (OA). We found that isopsoralen pretreatment significantly reversed the increase in DNA fragmentation and apoptosis rate, and significantly decreased the caspase-3 activity and PARP cleavage in IL-1ß-treated chondrocytes. Isopsoralen pretreatment markedly inhibited disruption of matrix proteins. Moreover, the expressions of LC3-II and LAMP-1 were markedly increased but the expression of p62/SQSTM1 was remarkably decreased by isopsoralen pretreatment. Importantly, the protective effects of isopsoralen against IL-1ß were blocked by pretreatment with autophagy inhibitor 3-MA and bafilomycin A1. These results suggest that isopsoralen ameliorates chondrocyte apoptosis by promoting autophagy flux.[Formula: see text].

The CtBP1-p300-FOXO3a transcriptional complex represses the expression of the apoptotic regulators Bax and Bim in human osteosarcoma cells.

C-terminal binding protein 1 (CtBP1), a well-known transcriptional corepressor, functions as an oncogene in multiple cancer types, including osteosarcoma, by modulating the transcription of many tumor suppressors, such as cadherin 1 (CDH1), phosphatase and tensin homolog (PTEN), Bcl2-associated X (Bax), Bcl-2-interacting mediator (Bim), and cyclin-dependent kinase inhibitor 1A (CDKN1A). However, it is still unclear how CtBP1 regulates the expression of these downstream targets. Here, we identified that CtBP1 is overexpressed in osteosarcoma cells and found that CtBP1 directly interacts with the transcription factor forkhead box O3 (FOXO3a) and the histone acetyltransferase p300 in vivo and in vitro. Through microarray analysis, we found that CtBP1 negatively regulates FOXO3a levels. In contrast to the CtBP1 level, the FOXO3a expression level was found to be significantly reduced in osteosarcoma cells. Knockdown of CtBP1 or overexpression of FOXO3a in U2OS cells resulted in different gene expression patterns, and the former caused upregulation of CtBP1 downstream target genes such as CDH1, PTEN, Bax, Bim, and CDKN1A, whereas the latter caused upregulation of Bax and Bim, but not CDH1, PTEN, and CDKN1A. Further analysis indicated that the CtBP1-p300-FOXO3a transcriptional complex specifically binds to the promoters of Bax and Bim. Inhibition of CtBP1 by the constitutive expression of Pep1-E1AWT peptide in U2OS and OSA cells reversed oncogenic phenotypes, including colony formation, cellular proliferation, and migration, and limited tumor growth in vivo. Together our results demonstrated that the CtBP1-p300-FOXO3a transcriptional complex represses the expression of the apoptotic regulators Bax and Bim in human osteosarcoma cells and that targeting CtBP1-mediated transcriptional events might be a potential therapeutic strategy for the osteosarcoma treatment.

Effect of methylene blue on outcomes in patients with distributive shock: a meta-analysis of randomised controlled trials.

OBJECTIVE: This meta-analysis aimed to demonstrate the effect of methylene blue (MB) in patients with distributive shock. DESIGN: Meta-analysis. METHODS: According to the Prospective International Register of Systematic Reviews (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines, we searched the relevant randomised controlled trials (RCTs) via PubMed, Embase and Cochrane Library from the date of database inception to 19 April 2023. The primary outcome was mortality during follow-up, and secondary outcomes included mean arterial pressure (mm Hg), mechanical ventilation time (hours), intensive care unit (ICU) length of stay (LOS) (days), hospital LOS (days) and heart rate (times/min). RESULTS: This study included six RCTs with 265 participants. The study showed no significant difference in mortality between the MB and placebo groups (ORs: 0.59; 95% CI 0.32 to -1.06). However, MB reduced the duration of mechanical ventilation (mean difference (MD): -0.68; 95% CI -1.23 to -0.14), ICU LOS (MD: -1.54; 95% CI -2.61 to -0.48) and hospital LOS (MD: -1.97; 95% CI -3.92 to -0.11). CONCLUSIONS: The use of MB may not reduce mortality in patients with distributive shock, but may shorten the duration of mechanical ventilation, ICU LOS and hospital LOS. More clinical studies are needed to confirm these findings in the future. TRIAL REGISTRATION NUMBER: CRD42023415938.

High prevalence of postoperative arytenoid dislocation in patients undergoing liver transplantation: A case-control study.

Arytenoid dislocation (AD) is a rare complication of surgery under general anesthesia. The potential factors for AD remain poorly defined, and the identification of risk factors is beneficial for reducing its incidence. We found that patients undergoing liver transplantation appeared to be more susceptible to postoperative AD at our hospital. The present study was designed to clarify this issue. A retrospective hospital-based case-control study was conducted in patients undergoing surgery under general anesthesia between 2017 and 2021. Recorded data for all patients were age, sex, body weight, height, body mass index, position of patients during surgery, duration of surgery, emergency status of surgery, and liver transplantation. Logistic regression analysis was performed to determine risk factors for AD. Thirty thousand one hundred fifty-four patients who underwent general anesthesia between 2017 and 2021 were included. Sixteen (0.05%) patients were diagnosed with AD, including 10 (3.9%) patients among 259 patients who underwent liver transplantation and 6 patients had complications among the 29,895 patients who underwent other operations (P < .0001). Postoperative AD incidence was significantly elevated in patients undergoing liver transplantation. This finding should be clinically relevant and alarming for anesthesiologists and clinicians to help avoid arytenoid dislocation and improve patient outcomes. Further studies that incorporate detailed data are needed to determine risk factors for AD.

Re-evaluation of serological criteria for early syphilis treatment efficacy: progression to neurosyphilis despite therapy.

OBJECTIVES: To study 17 cases of secondary syphilis that progressed to neurosyphilis despite appropriate treatments and whose rapid plasma reagin (RPR) titres showed a fourfold decrease within 6 months but did not revert to negative. METHODS: Secondary syphilis patients with the following criteria were analysed: (1) RPR titres declined fourfold within 3 months after therapy, (2) patients denied high-risk sexual behaviours following treatment, (3) RPR titre remained serofast 24 months after treatment, (4) reactive cerebrospinal fluid (CSF)-venereal disease research laboratory (VDRL) and CSF-Treponema pallidum Particle Agglutination Test (TPPA) and (5) HIV antibody negative. RESULTS: 14 male and three female patients met the criteria. 13 patients were asymptomatic. The CSF leucocyte count was elevated in 10 patients of whom nine also had elevated CSF-proteins. The RPR titres following secondary syphilis treatments were ≥ 1:32 in five cases, 1:16 in four cases, 1:8 in six cases and 1:4 in two cases. Following treatments for neurosyphilis, four cases with neurological or psychiatric manifestations resolved or improved, nine cases with raised CSF-white blood cells returned to normal and nine of 12 cases with raised CSF-protein declined to normal. CONCLUSIONS: Neurosyphilis may be detected in immunocompetent patients despite appropriate therapy for early-stage syphilis and appropriate serological responses. Clinicians should consider a CSF examination in any treated patient with evidence of disease progression irrespective of prior treatment history and serological response.

Azithromycin treatment failure among primary and secondary syphilis patients in Shanghai.

BACKGROUND: Azithromycin has been used to treat primary and secondary syphilis and as prophylaxis for sexual partners. We evaluated syphilis treatment failure in patients who received azithromycin therapy. METHODS: Patients who did not respond to azithromycin therapy were referred to Shanghai Skin Disease and sexually transmitted disease hospital. Treatment failure was defined as follows: (1) persistent ulcers or cutaneous or mucosal lesions 1 month after therapy; or (2) detection of spirochetes in dark-field microscopy examination of a lesion at least 1 week after treatment; or (3) failure of rapid plasma reagin titers to decrease 4-fold at 3 months after treatment. RESULTS: A total of 132 patients with primary and secondary syphilis who failed azithromycin therapy were referred to our hospital between January 2001 and October 2008. Of 132 patients, 42 (31.8%) had primary syphilis and 90 (68.2%) had secondary syphilis. Twenty-six patients with primary syphilis developed multiple lesions or secondary syphilis, or persistent ulcers despite using azithromycin. The skin or mucosal lesions did not resolve in 37 patients with secondary syphilis after azithromycin treatment. Ten patients had a positive dark-field examination for Treponema pallidum (T. pallidum) after treatment. The serum rapid plasma reagin titers studied in all cases had failed to decrease 4-fold at 3 months after therapy. The doses of azithromycin used for treatment ranged from 4 to 30 g. CONCLUSIONS: The failure of azithromycin to cure a substantial number of patients with primary and secondary syphilis in Shanghai suggests that azithromycin has limited therapeutic value in this setting.

Nonvenereal transmission of syphilis in infancy by mouth-to-mouth transfer of prechewed food.

Early recognition of acquired syphilis in childhood is vital. Children may acquire syphilis as a consequence of kissing, breast-feeding, or handling. We report 2 cases of infantile syphilis transmitted by mouth-to-mouth feeding from actively infected relatives. Syphilis should be suspected in children presenting with atypical rashes accompanied by headache, sore throat, and adenitis, especially if family members are affected by active syphilis.

Sensitive detection of Treponema pallidum DNA from the whole blood of patients with syphilis by the nested PCR assay.

The aim of this work was to investigate the application of the nested PCR assay for the detection of Treponema pallidum (TP) DNA from the blood of patients with different stages of syphilis. In this study, a nested PCR method targeting the Tpp47 and polA genes (Tpp47-Tp-PCR and polA-Tp-PCR) was developed to detect TP-DNA in whole blood samples collected from 262 patients with different stages of syphilis (84 primary syphilis, 97 secondary syphilis, and 81 latent syphilis patients). The PCR assay detected T. pallidum DNA in 53.6% and 62.9% of the patients with primary and secondary syphilis, respectively, which was much higher than the detection levels in patients with latent syphilis (7.4%) (both p < 0.001). For primary syphilis, a low RPR (0-16) was correlated with a higher detection rate of TP-DNA, whereas for secondary syphilis, the higher detection rate of blood TP-DNA was correlated with higher blood RPR titers (at or beyond 32). For latent syphilis, TP-DNA was only detectable by PCR in the early phase of the latent infection. Thus, blood RPR titers were correlated with the blood T. pallidum burden, but the correlations varied with primary and secondary syphilis. The results indicate that nested PCR is a sensitive method for detecting blood TP-DNA and is especially useful for detecting early syphilis including primary syphilis and secondary syphilis. The findings also suggest that the PCR assay may be used to complement other methods to enhance the diagnosis of syphilis.

Occurrence of congenital syphilis after maternal treatment with azithromycin during pregnancy.

OBJECTIVE: To evaluate the efficacy of azithromycin in preventing congenital syphilis. METHOD: Five pregnant women with syphilis who were allergic to penicillin were given azithromycin, 1 g daily orally or intravenously, in different hospitals. The duration of the therapy ranged from 1 day to 10 days. A second course of therapy was provided at 28 weeks gestation. The babies were given a physical examination and blood test for serum rapid plasma reagin test (RPR), treponema pallidum hemagglutination test (TPHA), and fluorescent treponemal antibody adsorption test (FTA-ABS-19-sIgM) within three months after birth. RESULTS: Five infants born to these mothers developed skin rashes. Four of the infants had hepatomegaly and one showed osteochondritis. The tests RPR, TPHA, and FTA-ABS-19-sIgM were positive. The RPR titers varied from 1:64 to 1:256 and the babies were diagnosed with congenital syphilis. They were successfully treated with penicillin. CONCLUSIONS: Successful therapy for syphilis during pregnancy demands maternal care as well as prevention or cure of congenital infection. The failure of azithromycin in preventing congenital syphilis in our report suggests that azithromycin should not be recommended as an alternative in treating syphilitic pregnant women or fetal syphilis.