OBJECTIVE: To elucidate the molecular mechanisms underlying sepsis-induced injury in mouse intestinal organoids and investigate the possible mechanisms or potential drug targets of myeloid differentiation factor 88 inhibitor [TJ-M2010-5 (TJ5)] on this condition. METHODS: Small intestinal organoids from C57BL/6 mice aged 6-8 weeks were established and characterized using immunofluorescence for cell growth and proliferation marker nuclear antigen Ki-67, goblet cell marker mucin-2 (MUC-2), epithelial cell marker E-cadherin, and Paneth cell marker lysozyme (Lyz). Small intestinal organoids after 3 days of passaging were divided into different groups: a normal control group treated with culture medium containing 0.2% dimethyl sulfoxide (DMSO) for 10 hours, a lipopolysaccharide (LPS) group treated with culture medium containing 200 mg/L LPS and 0.2% DMSO for 10 hours, and a TJ5 group pre-treated with 10 mmol/L TJ5 for 2 hours followed by treatment with culture medium containing 200 mg/L LPS for 10 hours. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to measure the expression levels of interleukin-6 (IL-6) and zonula occludens-1 (ZO-1) in the small intestinal organoids. RNA transcriptome sequencing was performed on the small intestinal organoids from each group to analyze differentially expressed genes between groups, and significant enrichment was analyzed using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). RESULTS: By the 7th day of primary culture, mature organoids had formed, and their growth rate increased after passaging. Immunofluorescence identification showed expressions of Ki-67, MUC-2, E-cadherin, and Lyz, indicating that the mouse small intestinal organoids maintained their cellular composition and functional characteristics under in vitro culture conditions. RT-qPCR results showed that compared with the normal control group, the mRNA expression of IL-6 in the small intestinal organoids of the LPS group was significantly increased (2^-ΔΔCT: 1.83±0.16 vs. 1.02±0.28, P < 0.05), while the mRNA expression of ZO-1 was significantly decreased (2^-ΔΔCT: 0.53±0.11 vs. 1.01±0.18, P < 0.05). In contrast, the mRNA expression trends of both IL-6 and ZO-1 were reversed in the TJ5 group, showing statistically significant differences as compared with the LPS group (2^-ΔΔCT: IL-6 mRNA was 1.24±0.01 vs. 1.83±0.16, ZO-1 mRNA was 1.97±0.29 vs. 0.53±0.11, both P < 0.05). RNA transcriptome sequencing showed 49 differentially expressed genes in the LPS group compared to the normal control group, with 42 upregulated and 7 downregulated. Compared to the LPS group, the TJ5 group showed 84 differentially expressed genes, with 47 upregulated and 37 downregulated. GO enrichment analysis of these differentially expressed genes showed that the significantly enriched biological processes of the differentially expressed genes between the normal control group and the LPS group included responses to LPS, responses to molecule of bacterial origin and responses to bacterium. The significantly enriched biological processes of the differentially expressed genes between the LPS group and the TJ5 group included glutathione metabolic processes, responses to stress cellular and responses to chemical stimulus. In molecular function groups, glutathione binding and oligopeptide binding were significantly enriched by the differentially expressed genes. In cellular component classifications, the enrichment of the differentially expressed genes was mainly observed in the cytoplasm, endoplasmic reticulum, and microsomes. KEGG pathway enrichment analysis indicated that the differentially expressed genes between the normal control group and LPS group were enriched in IL-17 signaling pathways, tumor necrosis factor (TNF) signaling pathways, viral protein interactions with cytokines and cytokine receptors signaling pathways, and cytokine-cytokine receptor interaction signaling pathways. In contrast, the differentially expressed genes between the LPS and TJ5 groups were mainly enriched in atherosclerosis signaling pathways, ferroptosis signaling pathways, glutathione metabolism signaling pathways, and cytochrome P450-mediated drug metabolism signaling pathways. CONCLUSIONS Mouse small intestinal organoids were successfully extracted and cultured. TJ5 may exert its protective effects by regulating gene expression and related signaling pathways (fluid shear stress and atherosclerosis, ferroptosis, glutathione metabolism, cytochrome P450 drug metabolism, etc.) in sepsis-injured mouse small intestinal organoids. These genes and signaling pathways may be key targets for treating sepsis-induced intestinal injury.
Animals ; Mice ; Sepsis/genetics* ; Organoids/drug effects* ; Mice, Inbred C57BL ; Intestine, Small/metabolism* ; Gene Expression Profiling ; Transcriptome ; Lipopolysaccharides

Objective:To analyze the effect of long-term care insurance on the incidence of falls in old people with disability, and the modification effect of different living arrangements.Methods:Based on the discharge diagnosis records of the hospitalization in urban residents who applied for long-term care insurance in Chengdu from July 2014 to June 2021, those who received long-term care insurance were selected as the intervention group and those who received no long-term care insurance were selected as the control group, the changes in the incidence of falls before and after the long-term care were analyzed using difference-in-differences method, and the modification effects of different living arrangements were explored using stratified analysis.Results:Long-term care insurance significantly reduced the risk for monthly incidence of falls in older adults [average treatment effects on treated (ATT) = -1.21% (95% CI: -1.82%--0.60%)], and the effect increased over time (ATT=-1.26%, 95% CI: -1.93%--0.59%). Long-term care insurance had a significant effect on reducing the incidence of falls in old people living with spouses/family members (ATT=-0.92%, 95% CI: -1.47%--0.37%) and in nursing facilities/hospitals (ATT=-2.10%, 95% CI: -3.81%--0.40%), but it had no significant effect in those living alone. Conclusions:Long-term care insurance can effectively reduce the incidence of falls in old people with disability, showing a positive effect. Living with family members or in nursing facilities/hospitals has a modification effect of the implementation of long-term care insurance policy. It is necessary to pay attention to key populations in the future and expand the coverage of long-term care insurance.

Objective:To study the current status of longitudinal extrauterine growth restriction (EUGR) in extremely preterm infants (EPIs) and to develop a prediction model based on clinical data from multiple NICUs.Methods:From January 2017 to December 2018, EPIs admitted to 32 NICUs in North China were retrospectively studied. Their general conditions, nutritional support, complications during hospitalization and weight changes were reviewed. Weight loss between birth and discharge > 1SD was defined as longitudinal EUGR. The EPIs were assigned into longitudinal EUGR group and non-EUGR group and their nutritional support and weight changes were compared. The EPIs were randomly assigned into the training dataset and the validation dataset with a ratio of 7∶3. Univariate Cox regression analysis and multiple regression analysis were used in the training dataset to select the independent predictive factors. The best-fitting Nomogram model predicting longitudinal EUGR was established based on Akaike Information Criterion. The model was evaluated for discrimination efficacy, calibration and clinical decision curve analysis.Results:A total of 436 EPIs were included in this study, with a mean gestational age of (26.9±0.9) weeks and a birth weight of (989±171) g. The incidence of longitudinal EUGR was 82.3%(359/436). Seven variables (birth weight Z-score, weight loss, weight growth velocity, the proportion of breast milk ≥75% within 3 d before discharge, invasive mechanical ventilation ≥7 d, maternal antenatal corticosteroids use and bronchopulmonary dysplasia) were selected to establish the prediction model. The area under the receiver operating characteristic curve of the training dataset and the validation dataset were 0.870 (95% CI 0.820-0.920) and 0.879 (95% CI 0.815-0.942), suggesting good discrimination efficacy. The calibration curve indicated a good fit of the model ( P>0.05). The decision curve analysis showed positive net benefits at all thresholds. Conclusions:Currently, EPIs have a high incidence of longitudinal EUGR. The prediction model is helpful for early identification and intervention for EPIs with higher risks of longitudinal EUGR. It is necessary to expand the sample size and conduct prospective studies to optimize and validate the prediction model in the future.

Objective:To investigate the effect of ultra-early enteral nutrition (UEEN) support on the prognosis of young and middle-aged postoperative patients with cerebral hemorrhage.Methods:The clinical data of young and middle-aged patients (aged 18-59 years) admitted to Tianjin Fifth Central Hospital from January 2020 to June 2023 after surgery for intracerebral hemorrhage were retrospectively analyzed, and the general data, nutritional indexes, gastrointestinal complications, neurological function recovery and long-term prognosis of the patients were recorded. According to the time of initiation of enteral nutrition (EN) support, patients were divided into UEEN group (EN implementation within 12 hour after surgery) and early enteral nutrition (EEN) group (EN implementation within 24 to 48 hour after surgery). The differences of the above indexes between the two groups were analyzed and compared.Results:A total of 64 young and middle-aged postoperative patients with cerebral hemorrhage were enrolled, including 32 cases in the UEEN group and 32 cases in the EEN group. There were no significant differences in gender, age, proportion of hypertension and diabetes, Glasgow coma score (GCS) on admission and surgical methods between the two groups. In terms of nutritional indexes, serum total protein, albumin and hemoglobin levels of patients in both groups on day 7 after admission were lower than those on day 1, and higher than those on day 3, and the above indexes levels in UEEN group were significantly higher than those in EEN group on day 7 [total protein (g/L): 63.05±5.79 vs. 59.02±6.63, albumin (g/L): 40.40±5.26 vs. 37.66±4.63, hemoglobin (g/L): 133.33±12.58 vs. 123.80±22.12, all P < 0.05]. In terms of gastrointestinal complications, the incidence of stress ulcer in the UEEN group within 14 days after admission was significantly lower than that in the EEN group [12.5% (4/32) vs. 31.3% (10/32), P < 0.05], but there was no statistically significant difference in feeding intolerance symptoms between the two groups. In terms of neurological recovery and long-term prognosis, GCS scores and Barthel index scores of 14 days after admission were higher than those of 1 day after admission, but there was no statistical significance between the two groups. Six months after surgery, Glasgow outcome scale (GOS) and Barthel index score of the UEEN group were significantly higher than those of the EEN group (GOS score: 3.81±1.06 vs. 3.18±1.07, Barthel index score: 60.78±7.24 vs. 54.52±5.13, both P < 0.05). Conclusion:UEEN support can improve the nutritional level of young and middle-aged postoperative patients with cerebral hemorrhage, reduce the occurrence of postoperative gastrointestinal complications, promote the recovery of neurological function, and improve the long-term prognosis.

Objective: To understand the public health emergency response capacity in primary and secondary schools, and to explore the problems and challenges in the prevention and control of public health emergency in primary and secondary schools for specific strategies. Methods: By using the stratified group sampling method, a questionnaire survey on general situation, knowledge, attitude and training, as well as public health emergencies response capacity among 2 988 teachers or leaders responsible for school emergency response in primary and secondary schools from Beijing, Chongqing and Yunnan. Results: Participants varied on their positions, titles, educational background and knowledge accuracy. Higher knowledge accuracy was associated with higher educational background ( χ 2=50.73-203.36, P < 0.05 ). The implementation of regular public health emergency related programs was poorly conducted in high schools (50.0%). Urban schools (42.0%) had higher proportion of qualified health care professionals than rural schools (18.2%), and private schools (48.5%) was higher than public schools (24.7%). The primary challenges included the shortage of guidance from professionals and the lack of related testing equipment (84.91%, 74.03%). Conclusion Although the ability of emergency handling of public health emergencies in schools in the three regions is advancing with the times, there are still many deficiencies, some omissions in the mastery of knowledge. It is suggested to inerease pre service and special training of school health work CDC should strengthen technical guidance and work supervision of infectious disease management in schools.

Objective:To explore the relationship between neutrophil to lymphocyte ratio(NLR) and first pass effect (FPE) during mechanical thrombectomy in patients with acute anterior circulation large vessel occlusive stroke.Methods:Four hundred and six patients with acute anterior circulation large vessel occlusive stroke, admitted to and received mechanical thrombectomy in our hospital from January 2018 to June 2021, were chosen in our study. They were divided into FPE group ( n=186) and non-FPE group ( n=220) according to whether the occluded vessels were successfully or completely re-canalized after the first mechanical thrombectomy. The baseline data, clinical characteristics and operation related data were recorded and compared. The factors with P<0.05 in univariate analysis were included in multivariate Logistic regression analysis to identify the independent factors for FPE. The receiver operating characteristic (ROC) curve was plotted to compare the effectiveness of various factors in predicting FPE. Results:Univariate analysis showed that there were significant differences in gender, proportion of different collateral circulation grading, neutrophil count, lymphocyte count, platelet to lymphocyte ratio (PLR), and NLR between the two groups ( P<0.05). As compared with non-FPE group, FPE group had significantly higher ASPECTS scores at admission, significantly shorter time from femoral artery puncture to recanalization, significantly higher proportion of vascular recanalization, statistically higher proportion of patients using balloon guiding catheter, significantly lower ratio of contrast extravasation, significantly lower incidences of spontaneous intracerebral hemorrhage and mortality, and statistically higher rate of good prognosis 90 d after surgery ( P<0.05). The results of multivariate Logistics regression analysis showed that gender ( OR=0.686, 95%CI: 1.131-3.491, P=0.017), ASPECTS scores ( OR=0.143, 95%CI: 0.094-0.220, P<0.001), NLR ( OR=1.722, 95%CI: 1.413-2.098, P<0.001), and PLR ( OR=1.007, 95%CI: 1.003-1.014, P<0.001) were independent factors for FPE. ROC curve results showed that the areas under the curve predicted by gender, ASPECTS scores, PLR, NLR and combination of multiple factors (gender+ASPECTS scores+PLR+NLR) were 0.60, 0.17, 0.71, 0.77 and 0.91, respectively; among them, NLR had the highest efficacy in single-factor prediction for FPE, with cut-off value of 5.86, specificity of 83.3%, and sensitivity of 62.3%. Conclusion:High NLR at admission in patients with acute anterior circulation large vessel occlusive stroke is likely to have FPE failure during mechanical thrombectomy.

Objective:To explore the characteristics of the changes in risk score for intensive care unit (ICU) patients during hospitalization by the intelligent calculation method, and to provide evidence for the risk prevention.Methods:In this retrospective study, ICU patients of the Fifth Central Hospital in Tianjin from November 3, 2021 to March 28, 2022 were enrolled and divided into ≥ 14 days group, 10-13 days group, 7-9 days group, and 3-6 days group according to the ICU length of stay. Risk scores assessed by the intelligent calculation method of the ICU patients were collected, including nutritional risk screening 2002 (NRS 2002), Caprini score and Padua score. NRS 2002 score for all patients, Caprini score for surgical patients and Padua score for internal medicine patients were selected. Trends in change of each score were compared between patients admitted to ICU 1, 3, 7 (if necessary), 10 (if necessary), and 14 days (if necessary).Results:A total of 138 patients were involved, including 79 males and 59 females, with an average age of (61.71±18.86) years and an average hospital stay of [6.00 (4.00, 9.25)] days. ① in the group with ICU length of stay ≥ 14 days (21 cases): there was no significant change in the NRS 2002 scores of the patients within 10 days, but the NRS 2002 score was significantly decreased in 14 days as compared with 1 day [3.00 (2.50, 3.50) vs. 4.00 (3.00, 5.00), P < 0.05]; both Caprini and Padua score were increased with prolonged hospital stay and compared with 1 day, the scores at the other time points were significantly increased, especially at 14 days [Caprini score: 5.00 (3.25, 7.00) vs. 2.50 (1.25, 5.50), Padua score: 6.00 (6.00, 7.00) vs. 3.00 (1.00, 3.00), both P < 0.05].② in the group with ICU length of stay from 10-13 days (15 cases): with the prolonged hospital stay, there was no significant change in NRS 2002 score, but both Caprini and Padua score were increased at 3, 7, 10 days, especially at 10 days [Caprini score: 3.00 (2.00, 4.75) vs. 2.00 (0.25, 2.75), Padua score: 5.00 (3.50, 6.00) vs. 2.00 (0.50, 4.00), both P < 0.05].③ in the group with ICU length of stay from 7-9 days (23 cases): compared with 1 day, the NRS 2002 score at 3 days and7 days were decreased, but the Caprini and Padua score were increased, especially at 7 days [NRS 2002 score: 2.00 (1.00, 4.00) vs. 2.00 (2.00, 4.00), Caprini score: 3.00 (2.00, 5.50) vs. 2.00 (0.25, 3.00), Padua score: 5.00 (4.00, 6.00) vs. 2.00 (0, 2.00), all P < 0.05]. ④ in the group with ICU length of stay from 3-6 days (79 cases): compared with 1 day, the NRS 2002 score at 3 days was decreased [NRS 2002 score: 2.00 (1.00, 3.00) vs. 2.00 (1.00, 3.00), P < 0.05], Caprini and Padua score were significantly increased [Caprini score: 3.00 (2.00, 4.00) vs. 2.00 (1.00, 3.00), Padua score: 5.00 (4.00, 5.00) vs. 2.00 (1.00, 3.00), both P < 0.05]. Conclusion:Based on dynamic assessment of intelligent calculation methods, the risk of thrombosis in ICU patients increased with hospital length of stay, and the nutritional risk was generally flat or reducing in different hospitalization periods.

Hepatotoxicity is a common side effect for patients treated with gefitinib, but the related pathogenesis is unclear and lacks effective predictor and management strategies. A multi-omics approach integrating pharmacometabolomics, pharmacokinetics and pharmacogenomics was employed in non-small cell lung cancer patients to identify the effective predictor for gefitinib-induced hepatotoxicity and explore optional therapy substitution. Here, we found that patients with rs4946935 AA, located in Forkhead Box O3 (FOXO3) which is a well-known autophagic regulator, had a higher risk of hepatotoxicity than those with the GA or GG variant (OR = 18.020, 95%CI = 2.473 to 459.1784, P = 0.018) in a gefitinib-concentration dependent pattern. Furthermore, functional experiments identified that rs4946935_A impaired the expression of FOXO3 by inhibiting the promotor activity, increasing the threshold of autophagy initiation and inhibiting the autophagic activity which contributed to gefitinib-induced liver injury. In contrast, erlotinib-induced liver injury was independent on the variant and expression levels of FOXO3. This study reveals that FOXO3 mutation, leading to autophagic imbalance, plays important role in gefitinib-induced hepatotoxicity, especially for patients with high concentration of gefitinib. In conclusion, FOXO3 mutation is an effective predictor and erlotinib might be an appropriately and well-tolerated treatment option for patients carrying rs4946935 AA.

Objective:To investigate the value of radiotherapy in patients with stage Ⅳ B thoracic esophageal squamous cell carcinoma (ESCC) at initial diagnosis. Methods:A total of 199 patients with stage Ⅳ B thoracic ESCC at initial diagnosis (according to UICC/AJCC Eighth Edition Esophageal and Esophagogastric Junction Cancer TNM Staging) who were treated in the Fourth Hospital of Hebei Medical University between January 2010 to December 2016 were recruited. Winthin the whole group, 130 patients (65.3%) had distant lymph node metastases alone, 51 cases (25.6%) of solid organ metastases alone and 18 cases (9.0%) of solid organ complicated with distant lymph node metastases. Among them, 16 patients (8.0%) were treated with chemotherapy alone, 50 cases (25.1%) of radiotherapy alone, 133 cases (66.8%) of radiochemotherapy (81 patients treated with concurrent radiochemotherapy and 52 patients treated with sequential radiochemotherapy). The survival rate was calculated by Kaplan-Meier method and the difference was analyzed by log-rank test. Clinical prognosis was assessed by multivariate Cox regression model. Results:The median overall survival (OS) of the entire cohort was 12.3 months (95% CI: 10.6-15.4m), and the 1-, 2-, 3-and 5-year OS rates were 52.1%, 25.2%, 19.1%, and 11.5%, respectively. Multivariate analysis showed that tumor length, the number of metastatic organs, and treatment modalities were the independent prognostic factors for OS. There was no significant difference in OS between concurrent radiochemotherapy and sequential radiochemotherapy ( P=0.955). The OS of patients in the radiotherapy dose of ≥6000 cGy group was significantly longer than that of their counterparts in the 4500-5039 cGy and 5040-6000 cGy groups (both P<0.001). Conclusions:For stage Ⅳ B thoracic ESCC patients at initial diagnosis, tumor length ≤3cm, single organ metastasis, and radiochemotherapy strategy are significantly correlated with longer OS. For stage Ⅳ ESCC patients with good physical status, radiotherapy can be supplemented on the basis of systemic chemotherapy. Concurrent or sequential radiochemotherapy needs to be individualized. If patients are tolerable, radiochemotherapy is recommended to the primary tumor or non-regional metastatic lymph nodes, aiming to prolong the OS of patients.

Objective:To compare the effects of freshwater and seawater drowning on sheep's pulmonary circulation hemodynamics and respiratory mechanics.Methods:According to the random number table method, healthy crossbred sheep were divided into freshwater drowning group ( n = 12) and seawater drowning group ( n = 12). 30 mL/kg of freshwater or seawater was infused respectively through trachea for approximately 5 minutes. Before the drowning, immediately after drowning, and 30, 60, 120 minutes after drowning, the systemic circulation hemodynamic parameters [heart rate (HR), mean arterial pressure (MAP), cardiac output (CO)] were monitored by pulse indicator continuous cardiac output (PiCCO); the respiratory parameters were obtained through the ventilator, including tidal volume (VT), lung compliance (Cdyn), oxygenation index (PaO 2/FiO 2), peak airway pressure (Ppeak)]; PiCCO and the right heart floating catheter (Swan-Ganz catheter) was used to measure pulmonary hemodynamic parameters [pulmonary systolic pressure (PAS), pulmonary diastolic pressure (PAD), pulmonary artery wedge pressure (PAWP), and extravascular lung water (EVLW)]. The animals were sacrificed at the end of the experiment, and the amount of residual water in the respiratory tract was measured; the pathological changes in the lung tissue were observed by hematoxylin-eosin (HE) staining. Results:① Systemic circulation hemodynamics: compared with the values before drowning, HR, MAP, and CO at the time of immediately after drowning in both freshwater and seawater were significantly increased and peaked. In addition, all indicators in the freshwater drowning group were significantly higher than those in the seawater drowning group [HR (bpm): 170.75±1.87 vs. 168.67±2.27, MAP (mmHg, 1 mmHg = 0.133 kPa): 172.92±1.62 vs. 159.42±3.18, CO (L/min): 13.27±0.71 vs. 10.33±0.73, all P < 0.05].② Respiratory parameters: compared with values before drowning, PaO 2/FiO 2, VT, and Cdyn decreased immediately in both freshwater and seawater drowning groups, Ppeak was significantly increased; in addition, the values in the seawater drowning group were decreased or increased more significantly than freshwater drowning group [PaO 2/FiO 2 (mmHg): 37.83±1.99 vs. 60.42±5.23, VT (mL): 86.25±7.66 vs. 278.75±9.67, Cdyn (mL/cmH 2O): 8.86±0.33 vs. 23.02±0.69, Ppeak (cmH 2O, 1 cmH 2O = 0.098 kPa): 42.17±2.69 vs. 17.67±1.15, all P < 0.01]. In addition, PaO 2/FiO 2 in the freshwater drowning group was gradually increased over time, while the seawater group continued to decline.③ Pulmonary circulation hemodynamic parameters: PAS, PAD, PAWP at the time of immediately after drowning in both freshwater and seawater groups were significantly higher than before drowning; in addition, the freshwater drowning group was significantly higher than the seawater drowning group [PAS (mmHg): 34.58±2.87 vs. 26.75±1.66, PAD (mmHg): 27.25±1.22 vs. 16.75±0.87, PAWP (mmHg): 27.83±1.85 vs. 11.75±1.82, all P < 0.01]. Thereafter, PAS and PAD in the freshwater drowning group gradually decreased, while the parameters in the seawater drown group continued to increase. PAWP gradually decreased after freshwater or seawater drowning, and recovered to pre-drowning levels 120 minutes after drowning and 30 minutes after drowning, respectively. EVLW continued to increase after freshwater drowning, reaching a peak at 30 minutes, and then decreased, until 120 minutes after drowning was still significantly higher than that before drowning (mL/kg: 10.73±1.27 vs. 7.67±0.69, P < 0.01); EVLW could not be measured.④ Residual water in the respiratory tract: residual water in the freshwater drowning group was significantly less than that in the seawater drowning group (mL: 164.33±25.21 vs. 557.33±45.23, P < 0.01).⑤ HE staining: partial alveolar atrophied in the freshwater drowning group, some alveolar spaces were broken, alveolar spaces and alveolar cavity showed a little powdery substance deposition; it was noted that alveolar expanded in the seawater drowning group, alveolar spaces were broken and bleeding and edema were obvious in the interstitial space. Conclusion:The effect of seawater drowning on the respiratory mechanics and pulmonary circulation of animals is more obvious than that of freshwater drowned animals, and the amount of residual water in the respiratory tract is also significantly more than that of freshwater drowned animals.