BACKGROUND:Ammonia poisoning is considered to be the main hypothesis for the pathogenesis of hepatic encephalopathy.Ammonia can lead to psychiatric and cognitive behavioral disorders,although the specific pathological molecular mechanisms remain unclear.OBJECTIVE:To investigate the effects of ammonia poisoning on cognitive behavior and hippocampal neuronal synapses in mice.METHODS:Thirty-two C57BL/6J mice were randomly divided into a normal control group and an ammonium chloride group,with 16 mice in each group.Normal saline was injected intraperitoneally in the control group,and ammonium chloride(10 mmol/kg)was injected intraperitoneally in the ammonium chloride group to construct a model of ammonia poisoning,once a day.After 7 days of ammonium chloride intervention,blood samples were collected from the hearts of six mice in each group for blood ammonia concentration detection.Behavioral experiments,including the open field test,novel object recognition test,and Y-maze test,were performed to assess mental and cognitive-behavioral changes in mice.Finally,hippocampal tissues were extracted for western blot analysis to detect the expression levels of synaptophysin and postsynaptic density protein-95 in hippocampal neurons.RESULTS AND CONCLUSION:The blood ammonia concentration was significantly elevated in the ammonium chloride group compared with the control group(P＜0.05).Mice in the ammonium chloride group showed anxiety-like behavior and disinhibition phenomenon,and a significant decrease in recognition memory and working memory ability.Western blot results revealed that the expression of synaptophysin and postsynaptic density protein-95 protein in hippocampal neurons in the ammonium chloride group was lower than that in the control group(P＜0.05).To conclude,ammonia poisoning can induce hippocampal neuronal synaptic damage,leading to psychiatric and cognitive behavioral abnormalities in mice.

BACKGROUND:Ammonia poisoning is considered to be the main hypothesis for the pathogenesis of hepatic encephalopathy.Ammonia can lead to psychiatric and cognitive behavioral disorders,although the specific pathological molecular mechanisms remain unclear.OBJECTIVE:To investigate the effects of ammonia poisoning on cognitive behavior and hippocampal neuronal synapses in mice.METHODS:Thirty-two C57BL/6J mice were randomly divided into a normal control group and an ammonium chloride group,with 16 mice in each group.Normal saline was injected intraperitoneally in the control group,and ammonium chloride(10 mmol/kg)was injected intraperitoneally in the ammonium chloride group to construct a model of ammonia poisoning,once a day.After 7 days of ammonium chloride intervention,blood samples were collected from the hearts of six mice in each group for blood ammonia concentration detection.Behavioral experiments,including the open field test,novel object recognition test,and Y-maze test,were performed to assess mental and cognitive-behavioral changes in mice.Finally,hippocampal tissues were extracted for western blot analysis to detect the expression levels of synaptophysin and postsynaptic density protein-95 in hippocampal neurons.RESULTS AND CONCLUSION:The blood ammonia concentration was significantly elevated in the ammonium chloride group compared with the control group(P＜0.05).Mice in the ammonium chloride group showed anxiety-like behavior and disinhibition phenomenon,and a significant decrease in recognition memory and working memory ability.Western blot results revealed that the expression of synaptophysin and postsynaptic density protein-95 protein in hippocampal neurons in the ammonium chloride group was lower than that in the control group(P＜0.05).To conclude,ammonia poisoning can induce hippocampal neuronal synaptic damage,leading to psychiatric and cognitive behavioral abnormalities in mice.

Objective:To establish a fluorescent quantitative RT-PCR assay based on N_sgRNA of SARS-CoV-2 and preliminarily apply it on real samples.Methods:Recombinant plasmid, specific primers and probes of N_sgRNA were designed and synthesized based on Wuhan-Hu-1/2019_MN908947 and synthesis mechanism of subgenomic RNA (sgRNA). Using recombinant plasmid as amplification templates, the optimal reaction conditions and reaction system were screened according to the Ct value, fluorescence intensity, and shape of amplification curve and was evaluated for sensitivity, reproducibility, and specificity. Meanwhile, the possibility of practical application of the method was explored by testing 172 clinical samples and 256 municipal wastewater samples. Results:A qRT-PCR assay for N_sgRNA in SARS-CoV-2 was initially established. The detection limit of the assay was 20 copies/mL, and the variation coefficients of in-batch (0.002%~0.767%) and batch to batch repetition (0.016%~0.752%) were less than 1%. Only N_sgRNA recombinant plasmid was detected in the specificity assay. So the method is more highly sensitive, specific and reproducible. The RatiosgRNA/ gRNA of clinical samples HK.3, EG.5.1, JN.1 and their sub-lineages and their corresponding urban sewage samples in epidemic period were significantly different ( P<0.05). There is a strong correlation between the median of RatiosgRNA/ gRNA in clinical samples and sewage samples in the same period (correlation coefficient r=1.000, P=0.010). Conclusions:In this study, a qRT-PCR method for detecting N_sgRNA of SARS-CoV-2 was established and the method has the characteristics of higher sensitivity, stronger specificity and better repeatability, and it can be used to detect SARS-CoV-2 infectivity.

Objective:The SARS-CoV-2 has a high natural mutation rate, and dynamic monitoring of virus variants remains a key focus in current COVID-19 prevention and control efforts.Methods:In this study, a sensitive and rapid method for detecting SARS-CoV-2 omicron variant nucleic acid was established based on the BMD-PCR technology.Results:This method showed good specificity, and had no cross-reactivity with 11 common viruses transmitted via the respiratory and gastrointestinal tracts, and the limit of detection is 555 copies/ml. Compared with SARS-CoV-2 whole-genome sequencing result, among 50 samples with original Ct values ≤32 tested for the Omicron variant, 49 samples tested positive for the N679K mutation site using BMD-PCR Omicron variant detection, achieving a concordance rate of 98.00%. For 30 samples JN.1 lineage, 29 samples tested positive for the K356T mutation site using BMD-PCR JN.1 lineage detection, with a concordance rate of 96.67%. For 10 samples with original SARS-CoV-2 detection Ct values between 35 and 32, 7 samples tested positive for the N679K mutation site using BMD-PCR Omicron variant detection, Resultsing in a detection rate of 70.00%. For samples with SARS-CoV-2 nucleic acid detection Ct values>35, the detection rate for the N679K mutation site in the BMD-PCR Omicron variant was 20.00%.Conclusions:This method can serve as a high-throughput supplementary approach for the preliminary identification of SARS-CoV-2 variant genotypes.

Objective:To investigate the prognostic impact of perineural invasion in patients with stageⅢ colon cancer and to clarify its guidance value for the duration of postoperative adjuvant chemotherapy.Methods:This study employed a retrospective cohort study method. It analyzed 426 patients with stageⅢ colon cancer who underwent radical surgery at Sun Yat-sen University Cancer Center and Longyan First Affiliated Hospital of Fujian Medical University, between April 2008 and June 2020. Inclusion criteria: patients received at least 3 months of adjuvant CapeOX therapy post-surgery, had complete pathological data, and were followed up for at least 12 months after the last chemotherapy. Among these patients, 231 were male, the median age was 59 (50~67) years, and 263 tumors were located in the right-sided colon. Postoperative pathology indicated that 107 cases (25.12%) had neural invasion, and 131 patients (30.75%) had vascular tumor thrombus. All patients received at least 4 cycles of postoperative CapeOX adjuvant chemotherapy, with 193 patients receiving 8 cycles and 233 patients receiving 4 to 7 cycles of adjuvant chemotherapy. The study analyzed the impact of neural invasion status and the duration of adjuvant chemotherapy on disease-free survival (DFS). Furthermore, within subgroups stratified by different risk levels (referencing the criteria proposed by the IDEA study: high risk: T4, N2 or T4N2; low risk: T3N1) and different neural invasion statuses, the impact of the duration of adjuvant chemotherapy on prognosis was analyzed.Results:The median follow-up time for the entire cohort was 94.00 months (55.27-128.80 months). Multivariate Cox analysis indicated that pathological T stage T4 (HR = 2.457, 95%CI: 1.499-4.029, P<0.001) and postoperative pathological confirmation of perineural invasion (HR = 2.465, 95% CI: 1.519-4.000, P<0.001) were independent adverse prognostic factors for 5-year DFS. In the perineural invasion-positive group, the 5-year DFS for patients who received 8 cycles of postoperative adjuvant CapeOX chemotherapy was 86.90%, compared to 58.22% for those who received 4-7 cycles, with statistically significant differences (both P<0.05). In the perineural invasion-negative group, the 5-year DFS for patients who received 8 cycles was 88.66%, compared to 90.99% for those who received 4-7 cycles, with no statistically significant differences ( P=0.929). Among IDEA high-risk patients with perineural invasion, the 5-year DFS was 91.81% for those who received 8 cycles versus 50.66% for those who received 4-7 cycles, showing a statistically significant difference ( P=0.003). In IDEA high-risk patients without perineural invasion, the 5-year DFS for those who received 8 cycles was 82.28% compared to 87.32% for those who received 4-7 cycles, with no statistically significant difference ( P=0.806). In the IDEA low-risk patients, no differences were observed in the 5-year DFS between patients receiving 8 cycles and those receiving 4-7 cycles of adjuvant CapeOX chemotherapy in both perineural invasion-positive and negative subgroups (both P>0.05). Conclusion:Perineural invasion serves as a significant prognostic factor for 5-year DFS in stage Ⅲ colon cancer patients who have undergone radical surgery and postoperative adjuvant chemotherapy. It can also be considered an important reference factor in deciding the duration of postoperative adjuvant chemotherapy.

Objective To systematically evaluate the risk prediction models for neonatal early-onset sepsis(EOS),aiming to provide reference for the construction and optimization of models,as well as for clinical selection of appro-priate prediction models.Methods PubMed,Web of Science,Embase,Cochrane Library,China National Know-ledge Infrastructure(CNKI),Wanfang Data,China Biology Medicine disc(CBM),and VIP databases were re-trieved,and studies relevant to neonatal EOS risk prediction models were collected.The retrieval period was from the inception of the database to January 18,2025.Two researchers independently screened literatures,extracted da-ta,and evaluated the quality of the included literatures using PROBAST tool.Any disagreements were resolved through consultation with a third reviewer.Results A total of 14 literatures were included in analysis,containing 19 risk prediction models.The area under receiver operating characteristic(ROC)curve(AUC)of the included model ranged 0.71-0.999.The number of prediction factors ranged 3-21.Common prediction factors included young gestational age,low birth weight,1-minute Apgar score,abnormal neonatal temperature,prolonged prema-ture rupture of membranes,amniotic fluid turbidity,maternal Group B streptococcal infection,maternal chorioam-nionitis,as well as elevated levels of procalcitonin and C-reactive protein in neonates.The risk of model overall bias was high,mainly due to insufficient number of outcome variable events in the analysis field,improper processing of missing data,screening of prediction factors based on univariate analysis,lacking model performance evaluation,and overfitting of model.Conclusion The neonatal EOS risk prediction model is still at the development stage.Al-though the current prediction models have better overall predictive performance,the overall quality needs to be im-proved.Future modeling can follow the PROBAST and TRIPOD specifications to reduce bias risk,explore the com-bination of multiple modeling methods,and focus on strengthening external validation and localized application to enhance the clinical applicability and promotion value of the model.

Objective:To investigate the relationship between glycated hemoglobin (HbA1c) level and the occurrence of diabetes complications in patients with type 2 diabetes mellitus in 11 provinces in China.Methods:A total of 4 832 patients with type 2 diabetes mellitus from 60 surveillance sites in 11 provinces where national surveillance for chronic diseases and risk factors was conducted in 2010 were selected as the study participants, and a follow-up survey was conducted in 3 516 persons from 2016 to 2017, finally 3 427 patients were included in the analysis after excluding those data exception and incomplete data. Cox proportional risk regression model was used to evalaute the association between HbA1c level and the risk for diabetes complications (macroangiopathy, microangiopathy and diabetic foot), and subgroup analyses were conducted according to the baseline characteristics of the study participants, such as age, gender and smoking status.Results:A total of 3 427 study participants were included in final analysis of the follow up for an average of 6.2 years, in whom 395 suffered from macroangiopathy, 226 suffered from microangiopathy, and 57 suffered from diabetic foot later during the follow-up period. After adjusting for relevant confounders, using the HbA1c <7.0% as a reference, there was no increased risk for macrovascular lesions in the those with HbA1c levels of 7.0%-, 7.5%-, 8.0%-8.4%, and the risk for macrovascular lesions increased by 38% in those with HbA1c ≥8.5% ( HR=1.38,95% CI:1.06-1.80); the risk for microangiopathies increased by 131% ( HR=2.31,95% CI:1.46-3.65), 206%( HR=3.06,95% CI:1.91-4.90) and 208% ( HR=3.08,95% CI:2.20-4.30) in those with HbA1c levels of 7.5%-, 8.0%-, ≥8.5%, respectively; and the risk for diabetic foot increased by 253% ( HR=3.53, 95% CI: 1.89-6.59) in those with HbA1c level ≥8.5%. Subgroup analyses revealed an effect modifying effect of different diabetes diagnosis situations (previously diagnosed and newly diagnosed) on HbA1c level and the risk for microangiopathy. Conclusions:HbA1c level ≥7.5% would increase the risk for microangiopathy in patients with type 2 diabetes mellitus, the higher the level, the higher the risk, and HbA1c level ≥8.5% would increase the risk for macrovascular lesions and diabetic foot. It is necessary to strengthen the health education in diabetic patients to improve their awareness of blood glucose management and the importance of HbA1c level control to effectively reduce or delay the diabetes complications.

Objective:To explore the current status and influencing factors of non-verbal communication needs in mechanically ventilated patients in ICU using an interpretive sequential mixed research design to inform the future development of targeted non-verbal communication strategies for mechanically ventilated patients in the ICU.Methods:Convenience sampling was used to select 262 mechanically ventilated patients from the general ICUs of two Class Ⅲ Grade A hospitals in Shanghai, from January to June 2023 for the study. Patients were surveyed using the General Information Questionnaire and the Surgical ICU Tracheal Intubation Patient Communication Needs Scale. Multiple linear regression was used to analyze the factors influencing the non-verbal communication needs of mechanically ventilated patients in the ICU. A total of 262 questionnaires were distributed in the quantitative study, and 256 valid questionnaires were recovered, with a valid recovery rate of 97.71% (256/262) . Purposive sampling was used to select 16 ICU mechanically ventilated patients for semi-structured in-depth interviews. The information was analyzed using the Colaizzi 7-step analysis method.Results:In 256 mechanically ventilated patients, the total non-verbal communication needs score was (144.33±12.82) , and the items average scores of physiological needs, safety needs, love and belongingness needs, and self-esteem needs were (3.39±1.83) , (3.35±0.98) , (3.32±1.21) , and (3.29±1.32) , respectively. Multiple linear regression analysis showed that the duration of mechanical ventilation, history of intubation, and education level were the factors influencing the non-verbal communication needs of mechanically ventilated patients in the ICU ( P<0.05) . Five themes were distilled from the qualitative study, including communication needs for shared decision-making about disease trajectories, communication needs for accurate management of disease symptoms, communication needs for psychological stress adjustment, communication needs for social system support, and communication needs for dignity preservation. Integrating and analyzing the quantitative and qualitative results revealed that they complemented each other in explaining and validating ideas in elaborating the current status and intrinsic relationship of non-verbal communication needs of mechanically ventilated patients in ICU. Conclusions:The non-verbal communication needs of mechanically ventilated patients in the ICU are prevalent and influenced by a variety of factors. It is recommended that hospital administrators construct an effective non-verbal communication support system based on the status quo of patients' non-verbal communication needs in order to promote the whole process and multidimensional health management services for ICU mechanically ventilated patients and to improve patients' quality of life.

BACKGROUND:Recent studies have found that Nrf2/ARE signaling pathway activators have the characteristics of low toxicity and control,and have a protective effect against radiation tissue damage.OBJECTIVE:To investigate whether nitrooleic acid can protect submandibular gland epithelial cells from radiation injury by regulating the Nrf2/ARE signaling pathway.METHODS:Rat submandibular gland epithelial cells were cultured in vitro and CCK-8 assay was used to screen the optimal concentration and time of nitrooleic acid administration.Submandibular gland epithelial cells were divided into non-radiation group,radiation control group,nitrooleic acid group,nitrooleic acid+ML385(Nrf2/ARE signaling pathway specific inhibitor)group,and ML385 group.Submandibular gland cells were pretreated with nitrooleic acid and ML385 for 24 hous according to the experimental groups,and then irradiated with 5 Gy radiation to establish the models.At 48 hours after irradiation,CCK-8 assay was used to detect the cell proliferation rate.Real-time quantitative PCR was used to detect the expression of Nrf2,HO-1,and NQO1 mRNA in the cells.Real-time quantitative PCR and enzyme-linked immunosorbent assay were used to detect the cell secretion function and the expression of inflammatory factors.DCFH-DA fluorescent probe kit was used to detect the level of intracellular reactive oxygen species.RESULTS AND CONCLUSION:(1)Compared with the radiation control group,the proliferation rate of submandibular gland epithelial cells and the expression levels of secretion function related factors aquaporin 5 and α-amylase in the nitrooleic acid group of rats increased(P＜0.05),and the expression levels of Nrf2,HO-1,and NQO1 mRNA increased(P＜0.05),while the expression levels of inflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factor-αdecreased(P＜0.05),and reactive oxygen species generation reduced(P＜0.01).(2)Compared with the nitrooleic acid group,the addition of nitrooleic acid and ML385 group resulted in a decrease in cell proliferation rate and expression levels of secretion function related factors aquaporin 5 and α-amylase(P＜0.05),and mRNA expressions of Nrf2,HO-1,and NQO1 were all decreased(P＜0.05),while the expression levels of inflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factor-α increased(P＜0.05),and generation of reactive oxygen species increased(P＜0.05).(3)Results indicated that in the radiation environment,nitrooleic acid has a certain protective effect on the proliferation ability and secretion function of rat submandibular gland epithelial cells,reduces the expression of inflammatory factors,lowers intracellular reactive oxygen species levels,and alleviates the damage of rat submandibular gland epithelial cells caused by radiation.This function may be related to the activation of Nrf2/ARE signaling pathway.

OBJECTIVE This study investigated the clinical implications of endothelial cell-specific molecule 1(ESM-1)in obstructive sleep apnea(OSA)patients,with particular focus on its dynamic correlation with adhesion molecules,aiming to elucidate the regulatory role of ESM-1 in OSA-associated vascular endothelial impairment.METHODS This cross-sectional study enrolled participants undergoing polysomnography(PSG)at the Sleep Medicine Center of Beijing Anzhen Hospital,Capital Medical University between March 2017 and January 2018.Based on the inclusion criteria,161 participants were ultimately included and divided into OSA group(n=118)and control group(n=43).Demographic data and polysomnography parameters were collected.We used a powerful high-throughput Multiplex Immunobead Assay technology to simultaneously test plasm cytokines levels of ESM-1,inter-cellular adhesion molecule 1(ICAM-1),vascular cell adhesion molecule 1(VCAM-1).Circulating C-reactive protein(CRP)and homocysteine(Hcy)were detected by routine blood chemistry panel.RESULTS Circulating ESM-1 levels were significantly elevated in patients with OSA compared with healthy controls[819.73(612.36-1393.47)pg/ml]vs.[286.17(114.48-513.81)pg/ml,P<0.001].After adjusting for confounding factors,we found that circulating ESM-1 levels were an independent risk factor for OSA(odds ratio=2.162,95%CI=1.522-3.072,P<0.001)and circulating ESM-1 levels were positively associated with ICAM-1 and VCAM-1 levels(β=1.977,95%CI=1.429-2.734,P<0.001).CONCLUSION Circulating ESM-1 levels were significantly increased in patients with OSA,which is closely related with adhesion molecules levels.ESM-1 may be a surrogate endothelial dysfunction marker and an independent risk factor for OSA.