Objective:To discuss the effect of asiatic acid(AA)on the inflammation and oxidative stress damage induced by lipopolysaccharide(LPS)in the primary cultured hippocampus neurons,and to clarify its mechanism.Methods:The primary cultured rat hippocampus neurons(cell purity identified by immunofluorescence staining)were divided into control group,LPS(10 mg·L-1)group,and LPS+AA group(10 mg·L-1 LPS+10,20,and 40 μmol·L-1 AA),AA group(20 μmol·L-1 AA),ML385 group[10 μmol·L-1 nuclear factor erythroid 2-related factor(Nrf2)inhibitor],and LPS+ML385+AA group(10mg·L-1 LPS+10 μmol·L-1 ML385+20 μmol·L-1 AA).After drug treatment,methylthiazolyldiphenyl-tetrazolium bromide(MTT)method was used to detect the survival rates of the hippocampus neurons in various groups;lactate dehydrogenase(LDH)kit was used to detect the LDH leakage rates of the hippocampus neurons in various groups;enzyme linked immunosorbent assay(ELISA)kit was used to detect the expression levels of inflammatory factors[interleukin(IL)-1β and tumor necrosis factor(TNF)-α]and the activities of superoxide dismutase(SOD)and malondialdehyde(MDA)levels in the hippocampus neurons in various groups;Griess method was used to detect the nitric oxide(NO)levels in supernatant of the hippocampus neurons in various groups;immunofluorescence staining was used to detect the expressions of Nrf2 and heme oxygenase-1(HO-1)proteins in the hippocampus neurons in various groups;Western blotting method was used to detect the expression levels of Nrf2,HO-1,nuclear factor-kappa B(NF-κB),and B-cell lymphoma 2(Bcl-2)proteins in the hippocampus neurons in various groups.Results:Compared with control group,the survival rate of the hippocampus neurons,SOD activity,and Bcl-2 expression level in the cells in LPS group were significantly decreased(P＜0.01),while the LDH leakage rate,expression levels of IL-1β and TNF-α,MDA level,and NO level,as well as the expression level of NF-κB protein,were significantly increased(P＜0.01);the fluorescence intensities and expression levels of Nrf2 and HO-1 proteins in hippocampus neurons were significantly decreased(P＜0.01).Compared with LPS group,the survival rates of hippocampus neurons,SOD activities,and expression levels of Bcl-2 in the cells in LPS+10 μmol·L-1 AA group and 20 μmol·L-1 AA group were significantly increased(P＜0.01),while the LDH leakage rates,expression levels of IL-1β and TNF-α,MDA levels,and NO levels,as well as expression levels of NF-κB protein,were significantly decreased(P＜0.05 or P＜0.01),and the fluorescence intensities and protein expression levels of Nrf2 and HO-1 in the cells were significantly increased(P＜0.01).Compared with LPS+20 μmol·L-1 AA group,the fluorescence intensities of Nrf2 and HO-1 in the cells in LPS+ML385+AA group were significantly decreased(P＜0.01),and the expression levels of Nrf2 protein in the nucleus and cytoplasm,the expression levels of HO-1 and Bcl-2 proteins in the cells were significantly decreased(P＜0.01),while the expression level of NF-κB protein was significantly increased(P＜0.01).Conclusion:AA can improve LPS-induced inflammation and oxidative stress damage in the primary cultured rat hippocampus neurons,and its mechanism may be related to the activation of the Nrf2/HO-1 signaling pathway.

Objective:To investigate symptoms of anxiety and depression among primary and middle school students quarantined in hotels during the COVID-19 epidemic.Methods:Anxiety and depression symptoms among 726 primary and middle school students quarantined in hotels were investigated with The Screen for Child Anxiety Related Emotional Disorders(SCARED)and Depression Self-rating Scale for Children(DSRSC)from September to October 2022 in Chifeng City,Inner Mongolia Autonomous Region.There were 624 students completed investi-gation with response rate of 86％.The positive score of SCARED was ≥23 and DSRSC was ≥ 15.Results:The detection rates of anxiety and depression were 17.9％and 15.4％respectively.The detection rates of anxiety and depression were higherin middle school students than inprimary school students(Ps＜0.05).The scores of general-ized anxiety and social phobia factors were higher in female students than in male students(Ps＜0.05).The scores of dissociative anxiety factor and depression were higher in middle school students than in primary school students(Ps＜0.05).Conclusion:During the COVID-19 epidemic,middle school students quarantined in hotels are more likely to have anxiety and depression symptoms than primary school students,and female students are more likely to have anxiety symptoms than male students.

Objective To improve the isolation and culture methods for primary hepatocytes and to establish a he-patic steatosis cell model for enhancing experimental efficiency and model precision.Methods Improvements were made upon existing techniques for primary hepatocyte cultivation and steatosis model establishment.The technology oriented to optimize the retrograde cannulation and fixation procedures,meticulously calibrating the perfusion speed and duration for mouse liver digestion.The culture medium was supplemented with 2% fetal bovine serum(FBS)during the removal of liver capsules and filtration steps.Additionally,the induction parameters for the steatosis cell model were refined,including the selection of free fatty acid(FFA)types and optimize their concentrations,ratios,and precise induction durations.Results The optimized protocol yielded mouse primary hepatocytes with a viability exceeding 90%,demonstrating ample quantity,favorable morphology,and excellent overall condition.The steatosis cell model exhibited prominent cytoplasmic lipid droplets,impaired glucose and lipid metabolism,as well as mild inflammation and insulin resistance,closely mimicking key aspects of the disease in vivo.Conclusions The refined techniques facilitated the establishment of a stable and physiologically representative in vitro steatosis cell model,which may support further research of pathogenesis of the disease,identification of potential therapeutic targets.

ACC oxidase (ACO) is one of the key enzymes that catalyze the synthesis of ethylene. Ethylene is involved in salt stress response in plants, and salt stress seriously affects the yield of peanut. In this study, AhACO genes were cloned and their functions were investigated with the aim to explore the biological function of AhACOs in salt stress response, and to provide genetic resources for the breeding of salt-tolerant varieties of peanut. AhACO1 and AhACO2 were amplified from the cDNA of salt-tolerant peanut mutant M29, respectively, and cloned into the plant expression vector pCAMBIA super1300. The recombinant plasmid was transformed into Huayu22 by pollen tube injection mediated by Agrobacterium tumefaciens. After harvest, the small slice cotyledon was separated from the kernel, and the positive seeds were screened by PCR. The expression of AhACO genes was analyzed by qRT-PCR, and the ethylene release was detected by capillary column gas chromatography. Transgenic seeds were sowed and then irrigated with NaCl solution, and the phenotypic changes of 21-day-seedings were recorded. The results showed that the growth of transgenic plants were better than that of the control group Huayu 22 upon salt stress, and the relative content of chlorophyll SPAD value and net photosynthetic rate (Pn) of transgenic peanuts were higher than those of the control group. In addition, the ethylene production of AhACO1 and AhACO2 transgenic plants were 2.79 and 1.87 times higher than that of control peanut, respectively. These results showed that AhACO1 and AhACO2 could significantly improve the salt stress tolerance of transgenic peanut.
Salt Tolerance/genetics* ; Arachis/genetics* ; Plant Breeding ; Ethylenes/metabolism* ; Plants, Genetically Modified/genetics* ; Gene Expression Regulation, Plant ; Plant Proteins/genetics*

Epilepsies are a group of chronic neurological disorders characterized by spontaneous recurrent seizures caused by abnormal synchronous firing of neurons and transient brain dysfunction. The underlying mechanisms are complex and not yet fully understood. Endoplasmic reticulum (ER) stress, as a condition of excessive accumulation of unfolded and/or misfolded proteins in the ER lumen, has been considered as a pathophysiological mechanism of epilepsy in recent years. ER stress can enhance the protein processing capacity of the ER to restore protein homeostasis through unfolded protein response, which may inhibit protein translation and promote misfolded protein degradation through the ubiquitin-proteasome system. However, persistent ER stress can also cause neuronal apoptosis and loss, which may aggravate the brain damage and epilepsy. This review has summarized the role of ER stress in the pathogenesis of genetic epilepsy.
Humans ; Endoplasmic Reticulum Stress/genetics* ; Unfolded Protein Response ; Endoplasmic Reticulum/pathology* ; Apoptosis ; Epilepsy/genetics*

Objective:To evaluate the effects of remimazolam on gastrointestinal motor function in the patients undergoing gastrointestinal endoscopy.Methods:A total of 262 American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 18-64 yr, with body mass index of 18-28 kg/m 2, scheduled for elective gastrointestinal endoscopy from May 2022 to August 2022, were divided into 2 groups ( n=131 each) using a random number table method: remimazolam group (group R) and propofol group (group P). The patients in group R received intravenous remimazolam 0.20-0.25 mg/kg, and patients in group P received intravenous propofol 1.5-2.0 mg/kg. The gastrointestinal endoscopy was performed when the patients′ Modified Observer′s Assessment of Alertness/Sedation scores ≤3. During fasting before gastrointestinal preparation, before gastrointestinal endoscopy and while leaving the post-anesthesia care unit (PACU), the concentrations of serum motilin and gastrin were measured by enzyme-linked immunosorbent assay, the intestinal peristalsis rating assessed by the endoscopist during the examination was recorded, the occurrence of hypotension and hypoxemia during the examination and occurrence of abdominal distension, abdominal pain, and nausea and vomiting during stay in PACU were recorded. Results:Compared with group P, the intestinal peristalsis rating was significantly increased, the serum motilin and gastrin concentrations were increased while leaving PACU, the incidence of hypotension and hypoxemia was decreased during the examination, and the incidence of abdominal distention was decreased during stay in PACU in group R ( P<0.05). Conclusions:Remimazolam has a milder inhibitory effect on secretion of gastrointestinal hormones than propofol in the patients undergoing gastrointestinal endoscopy and is helpful for the recovery of gastrointestinal motility.

Objective:To evaluate the obesity factor on ventilator-induced lung injury (VILI) in rats.Methods:Forty-five clean-grade male Sprague-Dawley rats, aged 6-8 weeks, were divided into 3 groups ( n = 15 each)according to the body weight: normal weight control group (group C), normal weight VILI group (group CV) and obese VILI group (group FV). The body weight was 233-267 g in C and CV groups and 288-332 g in FV group.In group C, the tidal volume (V T) was 10 ml/kg.In CV and FV groups, the rats were ventilated for 4 h with the V T set at 40 ml/kg, respiratory rate 40 breaths/min, inspiratory/expiratory ratio 1∶2, PEEP 0 mmHg, and fraction of inspired oxygen 21% to establish the VILI model.The arterial blood samples were collected immediately before tracheal intubation and at 4 h of mechanical ventilation for blood gas analysis and PaO 2 recording.The remaining blood samples were used for plasma collection.The rats were sacrificed after blood collection at 4 h of ventilation, and the bilateral lung tissues were isolated to collect the bronchoalveolar lavage fluid (BALF). The concentrations of leptin in plasma and tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and interleukin-6 (IL-6) in plasma and BALF were detected by enzyme-linked immunosorbent assay.The wet/dry weight (W/D) ratio of lung tissues was measured.The pathological changes of lung tissues were observed after HE staining, and the lung injury score was evaluated.The expression of NF-κB p65 in lung tissues was detected by Western blot. Results:Compared with group C and group CV, the plasma leptin concentration was significantly increased in group FV( P<0.01). Compared with group C, the concentrations of TNF-α, IL-6 and IL-1β in plasma and BALF were significantly increased, PaO 2 was decreased, the lung injury score and W/D ratio of lung tissues were increased, and NF-κB p65 expression was up-regulated at 4 h of ventilation in CV and FV groups ( P<0.01). Compared with group CV, the concentrations of TNF-α, IL-6 and IL-1β in plasma and BALF were significantly decreased, PaO 2 was increased, the lung injury score and W/D ratio of lung tissues were decreased, and NF-κB p65 expression was down-regulated at 4 h of ventilation in group FV ( P<0.05). Conclusion:Obesity factor can reduce VILI in rats, and the mechanism may be related to the increase in plasma leptin levels.

Objective To investigate the protective effect of microRNA-181b (miR-181b) on aged rats with sepsis-induced hippocampus injury in vivo. Methods Seventy-five male healthy old Sprague-Dawley (SD) rats were randomly divided into five groups (n = 15) using a random number table: sham operation group (Sham group), sepsis group [cecal ligation and puncture (CLP) group], miR-181b Agomir+CLP group (Ag+CLP group), miR-181b Antagomir+CLP group (An+CLP group) and normal saline (NS) control group (NS+CLP group). Rats sepsis model was reproduced by CLP, and in Sham group, the cecum of rats was separated only after abdominal operation without ligation or perforation. The rats in Ag+CLP group were given miR-181b Agomir 10 μL via lateral ventricle at 24 hours before CLP, the rats in An+CLP group were given 10 μL miR-181b Antagomir, and those in NS+CLP group were given 10 μL NS. At 6, 12, 24 hours after CLP, 5 rats of each group were sacrificed randomly, and hippocampus were harvested. The expression of miR-181b in hippocampus was determined by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR). The expression of nuclear factor-κB p65 (NF-κB p65) was determined by Western Blot. The contents of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). Results Compared with Sham group, the expression of miR-181b in hippocampus of CLP group was significantly decreased at 6 hours after CLP (2-ΔΔCT: 0.70±0.12 vs. 0.98±0.06, P < 0.05), and the expressions of NF-κB p65, IL-1β and TNF-α were significantly increased [NF-κB p65/Histone H3:0.30±0.03 vs. 0.07±0.01, IL-1β (ng/L): 120.39±8.02 vs. 50.55±11.12, TNF-α (ng/L): 59.48±4.60 vs. 40.31±3.96, all P < 0.05], this trend was continued till 24 hours, and these results indicated that there was obvious inflammation in hippocampus of sepsis rats. There was no statistical difference in the expression of miR-181b, NF-κB p65, IL-1β or TNF-α in hippocampus between NS+CLP group and CLP group, which indicated that injection of NS into the rat lateral ventricle, had not aggravated the damage degree of hippocampus. Compared with CLP group, the expression of miR-181b in hippocampus of Ag+CLP group was significantly increased at 6 hours after CLP (2-ΔΔCT: 1.87±0.25 vs. 0.70±0.12, P < 0.05), and the expressions of NF-κB p65, IL-1β and TNF-α were significantly lowered [NF-κB p65/Histone H3:0.16±0.03 vs. 0.30±0.03, IL-1β (ng/L): 73.76±8.17 vs. 120.39±8.02, TNF-α (ng/L): 49.52±4.77 vs. 59.48±4.60, all P < 0.05]. There was no statistical difference in the expression of miR-181b in hippocampus between An+CLP group and CLP group (2-ΔΔCT: 0.80±0.08 vs. 0.70±0.12 at 6 hours, 0.48±0.03 vs. 0.46±0.05 at 12 hours, 0.61±0.09 vs. 0.63±0.07 at 24 hours, all P > 0.05), but the expressions of NF-κB p65, IL-1β and TNF-α in hippocampus at 6 hours after CLP of An+CLP group were significantly higher than those of CLP group [NF-κB p65/Histone H3: 0.44±0.02 vs. 0.30±0.03, IL-1β (ng/L): 134.21±5.78 vs. 120.39±8.02, TNF-α (ng/L): 67.62±5.86 vs. 59.48±4.60, all P < 0.05], this trend was continued till 24 hours after CLP. The above results showed that overexpression of miR-181b might attenuate the inflammation of hippocampus through down-regulation of NF-κB, IL-1β and TNF-α. Conclusions The expression of hippocampal miR-181b was significantly decreased in septic rats. Up-regulation of miR-181b could inhibit the activation of NF-κB signal pathway and the release of the inflammatory cytokine IL-1β and TNF-α stimulated by sepsis, and alleviate the inflammatory reaction and hippocampus injury in rat with sepsis.

Objective:To investigate the significance of regulatory T cells in peripheral blood of chronic renal insufficientce.Methods:The peripheral blood samples were collected from 40 patients with chronic renal insufficient.The ratios ot CD4+T cell in lymphocyte and CD4+CD127-Treg and CD4+CD25+CD127-Treg in CD4+T were detected by flow cytometry.Results:The number of CD4+T in lymphocyte of chronic renal insufficient was higher than in healthy control group and there wasn’t significantly difference of the CD4+CD25+CD127-Treg ratios in CD4+T between chronic renal insufficience and healthy central.The ratio of CD4+T cells in lymphocytes of chronic renal insufficience was lower than in healthy control group except compensatory stage.There was no correlation between CD4+T cell ratios in lymphocytes,CD4+CD127-Treg or CD4+CD25+CD127-Treg ratios in CD4+T cells and the values of BUN,Cr among the hypertension patients.Conclusion:The number of CD4+T cells increases,and CD4+CD127-Treg decreases in the patients with chronic renal insufficience and their immune functions are shown in disoroler .