OBJECTIVE To prepare hyaluronic acid （HA）-modified emodin （EMD）-contained multi-walled carbon nanotubes （MWCNTs） drug delivery system （HA-MWCNTs-EMD） and explore its in vitro inhibitory effect on breast cancer cells. METHODS EMD was loaded onto MWCNTs to prepare a drug delivery system MWCNTs-EMD； subsequently， the system was further modified with HA to obtain the drug delivery system HA-MWCNTs-EMD. The two drug delivery systems mentioned above were characterized. With free EMD as the reference， the drug release in vitro of the above two drug delivery systems was investigated； the uptake of EMD by two breast cancer cells （MCF-7， MDA-MB-231 cells） was detected. The impacts of the above two drug delivery systems on the expression of surface glycoprotein differentiation group 44 （CD44）， activity， apoptosis and lactate dehydrogenase （LDH） release of two breast cancer cells were detected. RESULTS The encapsulation efficiencies of MWCNTs-EMD and HA-MWCNTs-EMD were both （63.52±2.74）%， with drug loading rates of （25.01±1.83）% and （12.13± 1.96）%， particle sizes of （865.95±2.16） and （351.86±1.68） nm， polydispersity indexes of 0.54±0.02 and 0.23±0.01， and Zeta potentials of （23.87±0.14） and （－42.79±0.39） mV， respectively. The 2， 4， 6， 8， 10， 12 and 24-hour cumulative release rates of EMD in MWCNTs-EMD and HA-MWCNTs-EMD were significantly lower than those in free EMD， while the cumulative release rate of HA-MWCNTs-EMD was significantly higher than that of MWCNTs-EMD （P＜0.05）； the EMD uptakes of MWCNTs-EMD and HA-MWCNTs-EMD by the two types of breast cancer cells were significantly higher than their uptake of free EMD （P＜0.05）. Compared with the free EMD group， the MWCNTs-EMD and MWCNTs-EMD groups showed significantly higher apoptosis rate and LDH release， significantly lower surface CD44 expression （except for the MWCNTs-EMD group） and cell viability in both cell types， and the effect of HA-MWCNTs-EMD was more pronounced （P＜0.05）. CONCLUSIONS A novel drug delivery system HA-MWCNTs- EMD loaded with EMD is developed successfully； the drug delivery system has a certain slow-release effect， which can significantly reduce the activity of breast cancer cells， promote their apoptosis and increase the release of LDH， and the above anti- breast cancer effect is significantly stronger than that of free EMD and MWCNTs-EMD.

Objective To explore the protective effect and mechanism of Jinyinqingre oral liquid on acute lung injury in-duced by lipopolysaccharide(LPS)in mice.Methods C57BL/6J mice were randomly divided into six groups according to the random number table method:blank control group,model control group,Jinyinqingre oral liquid low-dose group,Jinyinqingre oral liquid medium-dose group,Jinyinqingre oral liquid high-dose group,and dexamethasone group.Except for the blank control group,the other groups were injected with lipopolysac charide(LPS)(5 mg·kg-1)into the trachea to establish the acute lung injury model of mice,and the Jinyinqingre oral liquid low,medium,and high groups were continuously administered the drug by gavage for three days.After 24 h,lung tissue and bronchoalveolar lavage fluid(BALF)were collected from the six groups for follow-up detection.The pathological injury of lung tissue in each group was observed by HE staining.The total number of cells in BALF was detected.The to-tal protein content of BALF was detected by the BCA method.The contents of inflammatory cytokines TNF-α,IL-1β,IL-6,and IgM in BALF were detected by ELISA.The expression of NF-κB and NLRP3 proteins in lung tissues was detected by immunohistochemistry and Western blotting.Results Compared with model control group,Jinyinqingre oral liquid alleviated the pathological injury of lung tissue(P＜0.05),decreased the total cell count,total protein content and IgM expression in BALF(P＜0.01),and the expres-sion of inflammatory cytokines TNF-α,IL-6 and IL-1β in BALF was dreased(P＜0.05),the protein expressions of NF-κB and NL-RP3 in lung tissues was dreased(P＜0.05).Conclusion Jinyinqingre oral liquid attenuated the pathological injury,inflammatory exudation,and expression of inflammatory cytokines in LPS-induced lung injury in mice,and its mechanism may be through the reg-ulation of NF-κB/NLRP3 signaling pathway,providing a theoretical basis for its clinical application.

Objective:To investigate predictive factors for successful endovascular recanalization in patients with non-acute symptomatic internal carotid artery occlusion (SICAO), to develop a decision tree model using the Classification and Regression Tree (CART) algorithm, and to evaluate the predictive performance of the model.Methods:Patients with non-acute SICAO received endovascular therapy at 8 comprehensive stroke centers in China were included retrospectively. They were randomly assigned to a training set and a validation set. In the training set, the least absolute shrinkage and selection operator (LASSO) algorithm was used to screen important variables, and a decision tree prediction model was constructed based on CART algorithm. The model was evaluated using the receiver operating characteristic (ROC) curve, Hosmer-Lemeshow goodness-of-fit test and confusion matrix in the validation set.Results:A total of 511 patients with non-acute SICAO were included. They were randomly divided into a training set ( n=357) and a validation set ( n=154) in a 7:3 ratio. The successful recanalization rates after endovascular therapy were 58.8% and 58.4%, respectively. There was no statistically significant difference ( χ2=0.007, P=0.936). A CART decision tree model consisting of 5 variables, 5 layers and 9 classification rules was constructed using the six non-zero-coefficient variables selected by LASSO regression. The predictive factors for successful recanalization included fewer occluded segments, proximal tapered stump, ASITN/SIR collateral grading of 1-2, ischemic stroke, and a recent event to endovascular therapy time of 1-30 d. ROC analysis showed that the area under curve of the decision tree model in the training set was 0.810 (95% confidence interval 0.764-0.857), and the optimal cut-off value for predicting successful recanalization was 0.71. The area under curve in the validation set was 0.763 (95% confidence interval 0.687-0.839). The accuracy was 70.1%, precision was 81.4%, sensitivity was 63.3%, and specificity was 79.7%. The Hosmer-Lemeshow test in both groups showed P>0.05. Conclusion:Based on the type of ischemic event, the time from the latest event to endovascular therapy, proximal stump morphology, the number of occluded segments, and the ASITN/SIR collateral grading constructed the decision tree model can effectively predict successful recanalization after non-acute SICAO endovascular therapy.

Ectopic eruption of the first permanent molar is a common malocclusion in the early stage of mixed dentition. The incidence among children aged 6-10 years is 0.83%-8.7%, with an increasing trend. The main manifestation is mesial impaction of the first permanent molar during eruption, which is accompanied by the loss of root resorption and space loss of the second primary molar. Ectopic eruption of the first permanent molars can lead to underdevelopment of the dental arch and severe crowding of the posterior dentition. Prevention and early intervention are very important. The ectopic eruption of the first permanent molar is related to the disruption of the differential growth and developmental balance among the teeth, alveolar bone and jaw. In clinical practice, the diagnosis can be made according to the patient's medical history and clinical and imaging examinations, and the treatment can be performed by dividing the teeth, dividing the distal slices of the second primary molars, and distally moving the first permanent molars. This paper reviews the incidence, etiology, clinical manifestations, risks of ectopic eruption of the first permanent molar, diagnostic methods, and common treatment options to provide references for clinical treatment.

OBJECTIVE: To develop and validate a mechanical power (MP)-oriented nomogram prediction model of weaning failure in mechanically ventilated patients. METHODS: Patients who underwent invasive mechanical ventilation (IMV) for more than 24 hours and were weaned using a T-tube ventilation strategy were collected from the Medical Information Mart for Intensive Care-IV v1.0 (MIMIC-IV v1.0) database. Demographic information and comorbidities, respiratory mechanics parameters 4 hours before the first spontaneous breathing trial (SBT), laboratory parameters preceding the SBT, vital signs and blood gas analysis during SBT, length of intensive care unit (ICU) stay and IMV duration were collected and all eligible patients were enrolled into the model group. Lasso method was used to screen the risk factors affecting weaning outcomes, which were included in the multivariate Logistic regression analysis. R software was used to construct the nomogram prediction model and build the dynamic web page nomogram. The discrimination and accuracy of the nomogram were assessed by receiver operator characteristic curve (ROC curve) and calibration curves, and the clinical validity was assessed by decision curve analysis (DCA). The data of patients undergoing mechanical ventilation hospitalized in ICU of the First People's Hospital of Lianyungang City and the Second People's Hospital of Lianyungang City from November 2021 to October 2022 were prospectively collected to externally validate the model. RESULTS: A total of 3 695 mechanically ventilated patients were included in the model group, and the weaning failure rate was 38.5% (1 421/3 695). Lasso regression analysis finally screened out six variables, including positive end-expiratory pressure (PEEP), MP, dynamic lung compliance (Cdyn), inspired oxygen concentration (FiO₂), length of ICU stay and IMV duration, with coefficients of 0.144, 0.047, -0.032, 0.027, 0.090 and 0.098, respectively. Logistic regression analysis showed that the six variables were all independent risk factors for predicting weaning failure risk [odds ratio (OR) and 95% confidence interval (95%CI) were 1.155 (1.111-1.200), 1.048 (1.031-1.066), 0.968 (0.963-0.974), 1.028 (1.017-1.038), 1.095 (1.076-1.113), and 1.103 (1.070-1.137), all P < 0.01]. The MP-oriented nomogram prediction model of weaning failure in mechanically ventilated patients showed accurate discrimination both in the model group and external validation group, with area under the ROC curve (AUC) and 95%CI of 0.832 (0.819-0.845) and 0.879 (0.833-0.925), respectively. Furthermore, its predictive accuracy was significantly higher than that of individual indicators such as MP, Cdyn, and PEEP. Calibration curves showed good correlation between predicted and observed outcomes. DCA indicated that the nomogram model had high net benefits, and was clinically beneficial. CONCLUSIONS The MP-oriented nomogram prediction model of weaning failure accurately predicts the risk of weaning failure in mechanical ventilation patients and provides valuable information for clinicians making decisions on weaning.
Humans ; Respiration, Artificial/methods* ; Ventilator Weaning/methods* ; Nomograms ; Lung ; Risk Factors

BACKGROUND: Rheumatoid arthritis (RA), a chronic systemic autoimmune disease, is characterized by synovitis and progressive damage to the bone and cartilage of the joints, leading to disability and reduced quality of life. This study was a randomized clinical trial comparing the outcomes between withdrawal and dose reduction of tofacitinib in patients with RA who achieved sustained disease control. METHODS: The study was designed as a multicenter, open-label, randomized controlled trial. Eligible patients who were taking tofacitinib (5 mg twice daily) and had achieved sustained RA remission or low disease activity (disease activity score in 28 joints [DAS28] ≤3.2) for at least 3 months were enrolled at six centers in Shanghai, China. Patients were randomly assigned (1:1:1) to one of three treatment groups: continuation of tofacitinib (5 mg twice daily); reduction in tofacitinib dose (5 mg daily); and withdrawal of tofacitinib. Efficacy and safety were assessed up to 6 months. RESULTS: Overall, 122 eligible patients were enrolled, with 41 in the continuation group, 42 in the dose-reduction group, and 39 in the withdrawal group. After 6 months, the percentage of patients with a DAS28-erythrocyte sedimentation rate (ESR) of <3.2 was significantly lower in the withdrawal group than that in the reduction and continuation groups (20.5%, 64.3%, and 95.1%, respectively; P  < 0.0001 for both comparisons). The average flare-free time was 5.8 months for the continuation group, 4.7 months for the dose reduction group, and 2.4 months for the withdrawal group. CONCLUSION: Withdrawal of tofacitinib in patients with RA with stable disease control resulted in a rapid and significant loss of efficacy, while standard or reduced doses of tofacitinib maintained a favorable state. TRIAL REGISTRATION Chictr.org, ChiCTR2000039799.
Humans ; Quality of Life ; China ; Arthritis, Rheumatoid/drug therapy* ; Piperidines/therapeutic use* ; Treatment Outcome ; Antirheumatic Agents/therapeutic use* ; Pyrroles/therapeutic use*

Treating patients with esophageal squamous cell carcinoma (ESCC) is challenging due to the high chemoresistance. Growth differentiation factor 15 (GDF15) is crucial in the development of various types of tumors and negatively related to the prognosis of ESCC patients according to our previous research. In this study, the link between GDF15 and chemotherapy resistance in ESCC was further explored. The relationship between GDF15 and the chemotherapy response was investigated through in vitro and in vivo studies. ESCC patients with high levels of GDF15 expression showed an inferior chemotherapeutic response. GDF15 improved the tolerance of ESCC cell lines to low-dose cisplatin by regulating AKT phosphorylation via TGFBR2. Through an in vivo study, we further validated that the anti-GDF15 antibody improved the tumor inhibition effect of cisplatin. Metabolomics showed that GDF15 could alter cellular metabolism and enhance the expression of UGT1A. AKT and TGFBR2 inhibition resulted in the reversal of the GDF15-induced expression of UGT1A, indicating that TGFBR2-AKT pathway-dependent metabolic pathways were involved in the resistance of ESCC cells to cisplatin. The present investigation suggests that a high level of GDF15 expression leads to ESCC chemoresistance and that GDF15 can be targeted during chemotherapy, resulting in beneficial therapeutic outcomes.
Humans ; Esophageal Squamous Cell Carcinoma/drug therapy* ; Cisplatin/metabolism* ; Esophageal Neoplasms/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Carcinoma, Squamous Cell/genetics* ; Growth Differentiation Factor 15/therapeutic use* ; Receptor, Transforming Growth Factor-beta Type II/therapeutic use* ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic

Periodontitis is an inflammatory disease that is characterized by progressive destruction of the periodontium and causes tooth loss in adults.Periodontitis is known to be associated with dysbiosis of the oral microflora,which is often linked to various diseases.However,the complexity of plaque microbial communities of periodontitis,antibiotic resistance,and enhanced virulence make this disease difficult to treat.In this study,using metagenomic shotgun sequencing,we inves-tigated the etiology,antibiotic resistance genes(ARGs),and virulence genes(VirGs)of periodontitis.We revealed a significant shift in the composition of oral microbiota as well as several functional pathways that were represented significantly more abundantly in periodontitis patients than in con-trols.In addition,we observed several positively selected ARGs and VirGs with the Ka/Ks ratio＞1 by analyzing our data and a previous periodontitis dataset,indicating that ARGs and VirGs in oral microbiota may be subjected to positive selection.Moreover,5 of 12 positively selected ARGs and VirGs in periodontitis patients were found in the genomes of respiratory tract pathogens.Of note,91.8％of the background VirGs with at least one non-synonymous single-nucleotide polymorphism for natural selection were also from respiratory tract pathogens.These observations suggest a potential association between periodontitis and respiratory infection at the gene level.Our study enriches the knowledge of pathogens and functional pathways as well as the positive selection of antibiotic resistance and pathogen virulence in periodontitis patients,and provides evidence at the gene level for an association between periodontitis and respiratory infection.

Objective:To explore the diagnostic value of mechanical power (MP) in patients with moderate to severe acute respiratory distress syndrome (ARDS) based on the Medical Information Mart for Intensive Care-Ⅲv1.4 (MIMIC-Ⅲ v1.4).Methods:The information of ARDS patients undergoing invasive mechanical ventilation for no less than 48 hours who were hospitalized at Beth Israel Deaconess Medical Center in Boston, Massachusetts from June 2001 to October 2012 in the MIMIC-Ⅲ v1.4 were collected. The demographics of patients, disease severity scores, ARDS etiology, prognostic indicators, pre-ventilation arterial blood gas analysis and respiratory parameters within 48 hours of ventilation were extracted. According to the lowest oxygenation index (PaO 2/FiO 2) before ventilation, the patients were divided into mild to moderate ARDS group (> 150 mmHg, 1 mmHg≈0.133 kPa) and moderate to severe ARDS group (≤ 150 mmHg), and the differences in baseline characteristics between the two groups were compared. The independent predictors associated with the severity of ARDS were analyzed using Logistic regression. The receiver operator characteristic curve (ROC curve) was plotted. The area under ROC curve (AUC) was calculated to evaluate the diagnostic value of MP for moderate to severe ARDS. The Youden index was used to determine the diagnostic threshold of MP for moderate to severe ARDS. According to the cut-off value of MP based on Youden index, all ARDS patients were divided into high and low MP groups. Kaplan-Meier survival curve was used to analyze the 28-day survival status of patients. Results:A total of 403 ARDS patients were enrolled in the study, including 107 subjects with mild to moderate ARDS and 296 with moderate to severe ARDS. There were significant differences in age, sequential organ failure assessment (SOFA) score, the lowest PaO 2/FiO 2 before ventilation, the last PaO 2/FiO 2 before ventilation, 28-day mortality, the length of intensive care unit (ICU) stay, duration of mechanical ventilation, lung dynamic compliance (Cdyn) in the second 24 hours of ventilation and positive end-expiratory pressure (PEEP), plateau pressure (Pplat), driving pressure (ΔP), respiratory rate (RR), lung static compliance (Cst), MP, inspired fraction of oxygen (FiO 2) within 48 hours of ventilation between the two groups. After adjusting variables such as age, SOFA score, the last PaO 2/FiO 2 before ventilation, and related respiratory mechanics parameters, multivariate Logistic regression analysis showed that higher ΔP, PEEP and MP, and lower last PaO 2/FiO 2 before ventilation were independently associated with moderate to severe ARDS [odds ratio ( OR) and 95% confidence interval (95% CI) was 1.137 (1.032-1.252), 1.333 (1.139-1.561), 1.102 (1.030-1.179), and 0.996 (0.993-0.998), respectively, all P < 0.01]. The ROC curve analysis showed that the best cut-off value of MP for the diagnosis of moderate to severe ARDS was 18.1 J/min with sensitivity of 81.42% and specificity of 60.75%, and the AUC was 0.745 (95% CI was 0.690-0.799). According to the cut-off value of MP obtained by ROC curve, all ARDS patients were divided into high MP group (> 18.1 J/min) and low MP group (≤ 18.1 J/min). The Kaplan-Meier survival curve showed that the 28-day cumulative survival rate in the high MP group was significantly lower than that in the low MP group (73.8% vs. 85.1%; Log-Rank test: χ2 = 5.660, P = 0.017). Conclusion:MP is an independent predictor of the severity of ARDS, and it can be used to diagnose moderate to severe ARDS.

BACKGROUND: Sweat secreted by eccrine sweat glands is transported to the skin surface through the lumen. The eccrine sweat gland develops from the initial solid bud to the final gland structure with a lumen, but how the lumen is formed and the mechanism of lumen formation have not yet been fully elucidated. This study aimed to investigate the mechanism of lumen formation of eccrine gland organoids (EGOs). METHODS: Human eccrine sweat glands were isolated from the skin for tissue culture, and the primary cultured cells were collected and cultured in Matrigel for 14 days in vitro. EGOs at different development days were collected for hematoxylin and eosin (H&E) staining to observe morphological changes and for immunofluorescence staining of proliferation marker Ki67, cellular motility marker filamentous actin (F-actin), and autophagy marker LC3B. Western blotting was used to detect the expression of Ki67, F-actin, and LC3B. Moreover, apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) apoptosis assay kit, and the expression of poly (ADP-ribose) polymerase and Caspase-3 was detected by Western blot. In addition, 3-methyladenine (3MA) was used as an autophagy inhibitor to detect whether the formation of sweat glands can be effectively inhibited. RESULTS: The results showed that a single gland cell proliferated rapidly and formed EGOs on day 4. The earliest lumen formation was observed on day 6. From day 8 to day 14, the rate of lumen formation in EGOs increased significantly. The immunofluorescence and Western blot analyses showed that the expression of Ki67 gradually decreased with the increase in days, while the F-actin expression level did not change. Notably, the expression of autophagy marker LC3B was detected in the interior cells of EGOs as the apoptosis signal of EGOs was negative. Compared with the control group, the autophagy inhibitor 3MA can effectively limit the formation rate of the lumen and reduce the inner diameter of EGOs. CONCLUSION Using our model of eccrine gland 3D-reconstruction in Matrigel, we determined that autophagy rather than apoptosis plays a role in the lumen formation of EGOs.
Apoptosis ; Autophagy ; Eccrine Glands ; Epithelial Cells ; Humans ; Organoids