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.

Reactive oxygen species (ROS) are major contributors to radiation therapy-induced side effects in cancer patients. A fusion antioxidant enzyme comprising glutathione S-transferase (GST), superoxide dismutase 1 (SOD1), and a transmembrane peptide has been shown to effectively mitigate ROS-induced damage. To enhance its targeting capability, the fusion protein was further modified by incorporating a matrix metalloproteinase-2/9 substrate peptide (X) and the transmembrane peptide R9, yielding the antioxidant enzyme GST-SOD1-X-R9 (GS1XR). This modification reduced its transmembrane ability in tumor cells, thereby selectively protecting normal cells from oxidative stress. However, the use of non-human GST poses potential immunogenicity risks. In this study, we employed seamless cloning technology to construct an expression vector containing the human GST gene to replace the non-human GST gene, and then expressed and purified novel fusion antioxidant enzymes GS1R and GS1XR. The protective effects of newly constructed GS1R and GS1XR against hydrogen peroxide (H₂O₂)-induced oxidative damage in L-02 cells were then evaluated using GS1 as a control. Enzymatic activity assays revealed that the specific activity of GST in GS1XR remained unchanged compared to the unmodified protein, while SOD activity was enhanced. Exposure to 200 μmol/L H₂O₂ transiently activated the nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway; however, this activation diminished after 24 h, reducing cell viability to 48.4%. Both GS1R and GS1XR effectively scavenged intracellular ROS, directly counteracting oxidative stress and promoting Nrf2 nuclear translocation, thereby activating antioxidant pathways and restoring cell viability to normal levels. The two enzymes showed comparable efficacy. In contrast, GS1, lacking transmembrane capability, was restricted to scavenging extracellular ROS and provided only limited protection. In conclusion, both novel fusion antioxidant enzymes demonstrated significant potential in safeguarding normal cells from ROS-mediated oxidative damage. The findings provide a foundation for further investigation in related field.
Humans ; Oxidative Stress/drug effects* ; Hydrogen Peroxide ; Antioxidants/metabolism* ; Glutathione Transferase/metabolism* ; Recombinant Fusion Proteins/pharmacology* ; Superoxide Dismutase-1 ; Reactive Oxygen Species/metabolism* ; Superoxide Dismutase/biosynthesis*

Objective:To understand the occupational injury situation of front-line workers in metallurgical and shipbuilding and repairing industry, and explore the risk factors of occupational injury.Methods:From September 2023 to March 2024, using cluster sampling method, front-line workers from 2 metallurgical enterprises in Shaoguan and Jinan City and 2 shipbuilding and repairing enterprises in Jiangmen and Shenzhen City were selected as the investigation objects. 6248 questionnaires were distributed and collected, and 6178 were effective questionnaires, with a effective recovery rate of 98.88%. The basic information, living habits, working system, protection and occupational injury of workers were investigated, and the data of occupational injury in factories was collected. The types, jobs and main causes of occupational injuries in different industries were analyzed, and the influencing factors of occupational injuries were analyzed by univariate and multi-factor logistic regression.Results:The incidence of occupational injury was 3.13% (128/4086) in metallurgical industry and 4.02% (84/2092) in shipbuilding and repairing industry. The top three occupational injuries in the metallurgical industry were furnace worker (17.19%, 22/128), steel rolling worker (14.84%, 19/128), maintenance worker (10.16%, 13/128), and the top three injury types were mechanical injury (24.22%, 31/128), height fall (20.31%, 26/128) and object strikes (17.97%, 23/128). The top three occupational injuries in shipbuilding and repairing industry were welder (20.24%, 17/84), riveter (9.52%, 8/84) and crane (8.33%, 7/84). The top three injury types were hit by objects (34.52%, 29/84), hit by falling objects (22.62%, 19/84), and lifting injury (20.24%, 17/84). The injuries of workers in metallurgical industry and shipbuilding and repairing industry were mainly fractures, accounting for 32.03% (41/128) and 60.71% (51/84), respectively. The incidence of occupational injury was higher in males, with sleep disorder, high temperature exposure and chemical toxicity exposure ( P<0.05). There were significant differences in age, smoking degree, working age and emotional state between workers with occupational injury and those without occupational injury ( P<0.05). Multivariate analysis showed that male, age above 50 years old, moderate smoking, working years of 5-9 years, mild anxiety, poor health status and high temperature exposure were risk factors for occupational injury ( OR=25.57, 3.72, 14.27, 2.09, 1.50, 4.36, 0.66, P<0.05) . Conclusion:The incidence of occupational injury is higher in shipbuilding and repairing industry, and fracture is the main type of occupational injury. The occurrence of occupational injury is affected by gender, age, smoking, working age, emotional state, health status and high temperature exposure.

Objective:To understand the occupational injury situation of front-line workers in metallurgical and shipbuilding and repairing industry, and explore the risk factors of occupational injury.Methods:From September 2023 to March 2024, using cluster sampling method, front-line workers from 2 metallurgical enterprises in Shaoguan and Jinan City and 2 shipbuilding and repairing enterprises in Jiangmen and Shenzhen City were selected as the investigation objects. 6248 questionnaires were distributed and collected, and 6178 were effective questionnaires, with a effective recovery rate of 98.88%. The basic information, living habits, working system, protection and occupational injury of workers were investigated, and the data of occupational injury in factories was collected. The types, jobs and main causes of occupational injuries in different industries were analyzed, and the influencing factors of occupational injuries were analyzed by univariate and multi-factor logistic regression.Results:The incidence of occupational injury was 3.13% (128/4086) in metallurgical industry and 4.02% (84/2092) in shipbuilding and repairing industry. The top three occupational injuries in the metallurgical industry were furnace worker (17.19%, 22/128), steel rolling worker (14.84%, 19/128), maintenance worker (10.16%, 13/128), and the top three injury types were mechanical injury (24.22%, 31/128), height fall (20.31%, 26/128) and object strikes (17.97%, 23/128). The top three occupational injuries in shipbuilding and repairing industry were welder (20.24%, 17/84), riveter (9.52%, 8/84) and crane (8.33%, 7/84). The top three injury types were hit by objects (34.52%, 29/84), hit by falling objects (22.62%, 19/84), and lifting injury (20.24%, 17/84). The injuries of workers in metallurgical industry and shipbuilding and repairing industry were mainly fractures, accounting for 32.03% (41/128) and 60.71% (51/84), respectively. The incidence of occupational injury was higher in males, with sleep disorder, high temperature exposure and chemical toxicity exposure ( P<0.05). There were significant differences in age, smoking degree, working age and emotional state between workers with occupational injury and those without occupational injury ( P<0.05). Multivariate analysis showed that male, age above 50 years old, moderate smoking, working years of 5-9 years, mild anxiety, poor health status and high temperature exposure were risk factors for occupational injury ( OR=25.57, 3.72, 14.27, 2.09, 1.50, 4.36, 0.66, P<0.05) . Conclusion:The incidence of occupational injury is higher in shipbuilding and repairing industry, and fracture is the main type of occupational injury. The occurrence of occupational injury is affected by gender, age, smoking, working age, emotional state, health status and high temperature exposure.

Objective To evaluate the clinical value of free glycoprotein non-metastatic melanoma protein B(GPNMB)as a drug resis-tance and prognostic marker for non-small cell lung cancer(NSCLC)patients with epidermal growth factor receptor(EGFR)amplifica-tion accompanied by mutations.Methods Fifty-five cases of NSCLC patients with EGFR amplification associated with mutations who received treatment from March 2018 to September 2019 were included as the observation group.All patients received an EGFR-tyrosine kinase inhibitor(EGFR-TKI)as the first-line treatment;67 blood samples from the physical examination center during the same period were randomly included as healthy control.We compared the expression levels of free GPNMB between the two groups,explored the correlation between GPNMB expression and the clinicopathological information in the observation group;and combined the clinical efficacy to evaluate its value as a drug resistance marker.Through follow-up,the progress free survival(PFS)of patients was statistically analyzed,and through multivariate Cox regression analysis,independent risk factors affecting the survival in the observation group were explored.Results Compared with that in the control group,the expression level of free GPNMB in the observation group was signi-ficantly up-regulated.The expression level of free GPNMB in the observation group is significantly related to the clinical efficacy of EGFR-TKI(P = 0.016).Patients with high GPNMB expression have significantly stronger drug resistance,and patients with high GPNMB expression have significantly shorter PFS duration(P = 0.032).A high free GPNMB expression(HR = 4.029,95%CI:1.942-8.358,P＜0.001)is also an independent risk factor affecting patient survival.Conclusion The expression level of free GPNMB in patients with EGFR amplification accompanied by mutant NSCLC is significantly up-regulated,and its high expression is significantly related to the enhancement of the patient's drug resistance.High GPNMB expression is significantly related to the poor prognosis of patients and is an independent risk factor affecting patient survival.

OBJECTIVE: Identifying novel strategies to prevent particulate matter (PM)-induced lung injury is crucial for the reduction of the morbidity of chronic respiratory diseases. The combined intervention represented by herbal formulae for simultaneously targeting multiple pathological processes can provide a more beneficial effect than the single intervention. The aim of this paper is therefore to design a safe and effective medicinal and edible Chinese herbs (MECHs) formula against PM-induced lung injury. METHODS: PM-induced oxidative stress, inflammatory response and apoptosis A549 cell model were used to screen anti-oxidant, anti-inflammatory and anti-apoptotic MECHs, respectively. A network pharmacology method was utilized to rationally design a novel herbal formula. Ultra performance liquid chromatography-mass spectrometer was utilized to assess the quality control of MECHs formula. The excretion of magnetic iron oxide nanospheres of the MECHs formula was estimated in zebrafish. The MECH formula against PM-induced lung injury was investigated with mice experiments. RESULTS: Five selected herbs were rationally designed to form a new MECH formula, including Citri Exocarpium Rubrum (Juhong), Lablab Semen Album (Baibiandou), Atractylodis Macrocephalae Rhizoma (Baizhu), Mori Folium (Sangye) and Polygonati Odorati Rhizoma (Yuzhu). The formula effectively promoted the magnetic iron oxide nanospheres excretion in zebrafish. The mid/high dose formula significantly prevented PM-induced lung damage in mice by enhancing the activity of SOD and GSH-Px, reducing the MDA and ROS level and attenuating the upregulation of pro-inflammatory cytokine (IL-6, IL-8, IL-1β and TNF-α), down regulating the protein expression of NF-κB, STAT3 and Caspase-3. CONCLUSION Our findings suggest that the effective MECHs formula will become a novel strategy for preventing PM-induced lung injury and provide a paradigm for the development of functional foods using MECHs.

Objective:To explore whether BHLHE40 can affect the sensitivity of thyroid cancer (TC) cells to cisplatin by activating oxidative phosphorylation (OXPHOS) pathway by targeting high mobility group A2 (HMGA2) .Methods:The mRNA expression of HMGA2 and its upstream transcription factor BHLHE40 in TC tissues was analyzed by TCGA-THCA and hTFtarget online databases. The si-HMGA2, oe-HMGA2, oe-BHLHE40, negative control si-NC and oe-NC were transfected into TC cells (K1 and SW579) by liposome transfection method. The mRNA expression levels of BHLHE40 and HMGA2 in TC cells (SW579, FTC-133, and K1) and normal thyroid cells (Nthy ori3-1) were detected by real-time quantitative PCR (qRT-PCR). The cell viability was detected by MTT assay, the half inhibitory concentration (IC 50) value of cisplatin was calculated by CCK-8 assay, the apoptosis level was detected by flow cytometry, and the expression of OXPHOS complex was detected by Western blotting. Seahorse XFe 96 was used to analyze the oxygen consumption rate of the TC cells. Dual-luciferase assay and chromatin immunoprecipitation (ChIP) assay were used to analyze the binding relationship between BHLHE40 and HMGA2. Results:TCGA database results showed that the mRNA expression levels of HMGA2 and BHLHE40 in TC tissues (10.57±2.58, 13.89±1.13) were higher than those in normal thyroid tissues (4.82±1.69, 12.28±1.01), with statistically significant differences ( t=16.69, P<0.001; t=10.43, P<0.001). The results of qRT-PCR showed that the relative mRNA expression levels of HMGA2 in normal thyroid cells (Nthy ori3-1) and TC cells (SW579, FTC-133, and K1) were 1.00±0.13, 2.94±0.23, 4.71±0.41 and 6.29±0.49, while those of BHLHE40 were 1.00±0.12, 2.60±0.23, 3.39±0.35 and 6.18±0.51 respectively, both with statistically significant differences ( F=130.50, P<0.001; F=125.20, P<0.001). Further pairwise comparison showed that mRNA expression levels of HMGA2 and BHLHE40 in TC cells were significantly higher than those in normal thyroid cells (all P<0.001). According to MTT experimental results, si-HMGA2 treatment significantly reduced the cell viability of K1 cells compared to the si-NC group (all P<0.05). In addition, compared to the oe-NC group, oe-HMGA2 treatment significantly increased the cell viability of SW579 cells (all P<0.05). Compared to the oe-NC+DMSO group, the oe-HMGA2+DMSO group showed enhanced cell viability of SW579 cells, while the OXPHOS pathway inhibitor Gboxin was able to reverse the effect of overexpressing HMGA2 on cell viability (all P<0.05). The results of flow cytometry and CCK-8 experiments showed that compared to the si-NC group (apoptosis level: 6.19%±0.28%; cisplatin IC 50 value: 17.47 μmol/L), knocking down HMGA2 could increase the apoptosis level (11.96%±0.32%; t=19.17, P<0.001) and cisplatin sensitivity (IC 50 value: 1.49 μmol/L) of K1 cells. In addition, compared to the oe-NC group (apoptosis level: 9.98%±0.32%; cisplatin IC 50 value: 8.17 μmol/L), overexpressing HMGA2 significantly decreased the apoptosis level (4.32%±0.25%; t=19.65, P<0.001) and cisplatin sensitivity (IC 50 value: 34.95 μmol/L) of SW579 cells. The results of dual-luciferase assay showed that compared with the si-NC group, knocking down the expression of BHLHE40 in human kidney epithelial 293T cells significantly reduced the luciferase activity of wild-type HMGA2 (0.31±0.02 vs. 1.00±0.11; t=10.69, P=0.004). However, there was no significant effect on the luciferase activity of mutant-type HMGA2 (1.06±0.11 vs. 1.00±0.07; t=0.80, P=0.470). ChIP results showed that the mRNA expression level of HMGA2 in K1 cells was significantly increased in the anti-BHLHE40 group (6.57±0.62) compared with the IgG group (1.00±0.10; t=15.36, P<0.001). Compared to the oe-NC+DMSO group, the oe-HMGA2+DMSO group showed decreased apoptosis level ( P<0.05) and cisplatin sensitivity of SW579 cells, with a significant increase in the expression of OXPHOS complexes Ⅰ-Ⅴ and cellular oxygen consumption rates (all P<0.05). The effect of overexpressing HMGA2 was reversed by treatment with oe-HMGA2+Gboxin (all P<0.05). The recovery experiment showed that compared to the oe-NC+si-NC group, overexpression of BHLHE40 in SW579 cells increased cell viability and the expression of OXPHOS complexes Ⅰ-Ⅴ, while decreasing apoptosis levels and increasing cellular oxygen consumption rates and cisplatin IC 50 values (all P<0.05). However, simultaneous knockdown of HMGA2 reversed the effect of overexpressing BHLHE40 (all P<0.05) . Conclusion:BHLHE40 can activate the OXPHOS pathway by targeting and regulating the expression of HMGA2, thereby affecting the sensitivity of TC cells to cisplatin.

Objective:To study the clinical value of lung ultrasound score (LUSsc) within 2 h after birth for pulmonary surfactant (PS) use in preterm infants with respiratory distress syndrome (RDS).Methods:From July 2019 to May 2021, preterm infants with RDS hospitalized in our hospital and received pulmonary ultrasound within 2 h after birth were prospectively enrolled. 12-area LUSsc was calculated. The infants were assigned into <32 weeks group and 32-36 weeks group according to gestational age (GA). Simple random sampling was carried out in each group with 1/5 as the validation set and the other 4/5 as the training set. The infants were also assigned into PS group and non-PS group according to PS usage within 24 h after birth. Receiver operator characteristic (ROC) curve of LUSsc predicting PS usage was drawn and validated.Results:A total of 857 RDS infants were enrolled, including 313 in <32 weeks group and 544 in 32-36 weeks group. For <32 weeks group, area under curve (AUC) of LUSsc>8.5 predicting PS use was 0.779 (95% CI 0.722-0.837), with 76.4% sensitivity and 81.4% specificity. The accuracy of using LUSsc>8.5 as cut-off predicting actual clinical PS application was 82.3% (Kappa value 0.692, P<0.05, McNemar's test P>0.05).For 32-36 weeks group, AUC of LUSsc>9.5 predicting PS use was 0.785 (95% CI 0.723-0.848), with 71.1% sensitivity and 81.7% specificity. The accuracy of using LUSsc>9.5 as cut-off predicting actual clinical PS application was 92.6% (Kappa value 0.772, P<0.05, McNemar's test P>0.05). Conclusions:LUSsc within 2 h after birth is independent predictor of PS use in preterm infants with RDS. For <32 weeks group, LUSsc>8.5 suggests PS application and for 32-36 weeks group the cut-off is LUSsc>9.5.

Objective:To explore the clinical features of patients with syndrome of intracranial hypotension (SIH) complicated by bilateral chronic subdural hematoma (CSDH).Methods:A case-control study was conducted; 16 patients with SIH complicated with bilateral CSDH (SIH group) and 32 patients with bilateral CSDH (non-SIH group) admitted to Department of Neurology and Neurosurgery, Shanghai Tenth People's Hospital Affiliated to Tongji University from January 2016 to October 2020 were selected. The differences of demographic characteristics, initial symptoms, medical history and CT image features between the two groups were compared.Results:(1) In 16 patients from the SIH group, 13 (81.3%) complained of typical postural headache symptoms, 3 (18.6%) showed fake subarachnoid hemorrhage on CT, 80.0% (12/15) showed dural diffuse enhancement on MRI, and 33.3% (5/15) showed signs of brain droop. Spinal MRI showed 27.3% patients (3/11) had signs of intracranial cerebrospinal fluid leakage. Of the 10 patients underwent bilateral trepanation and drainage, 6 experienced postoperative deterioration (4 received multiple additional surgeries including decompressive craniectomy, and 1 severe patient died in hospital after giving up treatment due to malignant tumor). (2) SIH group had significantly younger age, and significantly lower percentages of patients with limb weakness symptoms, hypertension, head trauma histories and increased hematoma pressure during trepanation and drainage, significantly lower age-adjusted comorbidities index, significantly decreased total and differential thickness of bilateral hematoma on CT, significantly shorter disease course, and statistically higher proportion of patients with postural headache and hematoma uniform density than non-SIH group ( P<0.05). Conclusion:According to age, initial symptoms and CT features, bilateral CSDH patients caused by SIH can be identified to a certain extent, and cranial and spinal MRI is recommended for definitive diagnosis of SIH.