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.

Methods: This retrospective study analyzed 32 consecutive IDS patients who underwent UEDD surgery. Clinical features, laboratory data (erythrocyte sedimentation rate and C-reactive protein), and treatment outcomes were analyzed. Results: Definite microorganisms were identified in 27 patients (84.3%), with 24 (88.9%) meeting cure criteria. The cure rate was significantly higher in the detected pathogen group compared to the undetected pathogen group (88.9% vs. 80%; χ²=19.36, p<0.0001). Metagenomic next generation sequencing (mNGS) provided faster diagnosis (41.72±6.81 hours) compared to tissue culture (95.74±35.47 hours, p<0.05). The predominant causative pathogen was Mycobacterium tuberculosis, followed by Staphylococcus aureus. Significant improvements were observed in Visual Analog Scale pain scores, from a mean of 7.9 preoperatively to 1.06 at 1 year postoperatively. The Oswestry Disability Index revealed a similar trend, showing significant improvement (p<0.05). Conclusions UEDD is a viable alternative to traditional open surgery for managing IDS in high-risk patients. UEDD offers a dual therapeutic-diagnostic advantage during the initial admission phase, enabling simultaneous debridement, neurological decompression, and targeted biopsy in a single intervention. Compared with traditional tissue culture, mNGS enables rapid microbiological diagnosis and extensive pathogen coverage.

Objective:The variability of the apnea-hypopnea index（AHI） measured in the first and second halves of the night is significant in patients with obstructive sleep apnea hypopnea syndrome（OSAHS）. This variation may be related to fluid redistribution caused by the supine position during sleep. Methods:Eighty-nine adult subjects were enrolled. Circumferences（neck, chest, waist, and calf） were measured before sleep onset and upon awakening. Polysomnography（PSG） was performed, and the night was divided into two halves based on the midpoint of total sleep time to calculate AHI for each half. The correlation between changes in AHI and changes in circumferences was analyzed. Results:Twenty simple snorers and sixty-nine OSAHS patients were included, with a median AHI of 22.6（11.8, 47.3） events/hour. Compared to pre-sleep measurements, there was no significant change in neck circumference upon awakening in the control group（P=0.073）, while reductions were observed in the other three measurements（P=0.006, P=0.038, P<0.001）. In the OSAHS group, neck circumference increased（P<0.001）, and reductions were noted in the other three measurements（P<0.001 for all）, with the most significant change observed in calf circumference 40.0（37.1, 42.0） cm to 38.0（35.8, 40.5） cm. Compared to the first half of the night, total AHI, supine AHI, and NREM AHI significantly decreased in the second half（P=0.010, P=0.031, P=0.001）, while no significant changes were observed in lateral AHI and REM AHI（P=0.988, P=0.530）. Further analysis revealed a significant relationship between increased chest circumference and decreases in NREM AHI, supine AHI, and supine NREM AHI（P=0.036, P=0.072, P=0.034）, as well as between decreased lateral position AHI and increased waist circumference（P=0.048）. Additionally, this study found a negative correlation between changes in calf circumference and changes in AHI（R=-0.24, P=0.048）, while neck circumference changes positively correlated with changes in AHI（R=0.26, P=0.03）. Conclusion:In OSAHS patients during the second half of sleep compared to before sleeping, chest circumference, waist circumference, and calf circumference decrease while neck circumference increases; total AHI, supine position AHI, and NREM period AHI decrease; increases in chest circumference are associated with decreases in NREM period AHI, supine position AHI, supine position NREM period AHI. There is nocturnal variability in AHI among OSAHS patients that may be associated with fluid shifts during sleep.
Humans ; Sleep Apnea, Obstructive/physiopathology* ; Male ; Female ; Polysomnography ; Fluid Shifts/physiology* ; Adult ; Middle Aged ; Neck ; Severity of Illness Index ; Sleep/physiology* ; Snoring/physiopathology*

Anxiety disorder is a major symptom of autism spectrum disorder (ASD) with a comorbidity rate of ~40%. However, the neural mechanisms of the emergence of anxiety in ASD remain unclear. In our study, we found that hyperactivity of basolateral amygdala (BLA) pyramidal neurons (PNs) in Shank3 InsG3680 knock-in (InsG3680^+/+) mice is involved in the development of anxiety. Electrophysiological results also showed increased excitatory input and decreased inhibitory input in BLA PNs. Chemogenetic inhibition of the excitability of PNs in the BLA rescued the anxiety phenotype of InsG3680^+/+ mice. Further study found that the diminished control of the BLA by medial prefrontal cortex (mPFC) and optogenetic activation of the mPFC-BLA pathway also had a rescue effect, which increased the feedforward inhibition of the BLA. Taken together, our results suggest that hyperactivity of the BLA and alteration of the mPFC-BLA circuitry are involved in anxiety in InsG3680^+/+ mice.
Animals ; Prefrontal Cortex/metabolism* ; Basolateral Nuclear Complex/metabolism* ; Mice ; Anxiety/metabolism* ; Nerve Tissue Proteins/genetics* ; Male ; Gene Knock-In Techniques ; Pyramidal Cells/physiology* ; Mice, Transgenic ; Neural Pathways/physiopathology* ; Mice, Inbred C57BL ; Microfilament Proteins

This study reports the diagnosis and treatment of six cases of post-transplant lymphoproliferative disorder (PTLD) in liver transplant recipients, confirmed at the Affiliated Hospital of Qingdao University between August 2017 and May 2023. The report includes details on anti-rejection therapy, Epstein-Barr virus (EBV) and cytomegalovirus (CMV) infections, imaging findings, histopathological results, treatment courses, and prognoses. By summarizing the clinical experience in the diagnosis and management of PTLD following liver transplantation, this study aims to provide valuable insights and references for the clinical diagnosis and treatment of this condition.

Objective:To investigate the effects of musculoskeletal ultrasound(MSUS)-guided shoulder joint capsule hydraulic expansion method combined with rotator interval(RI)injection method on ultrasound imaging indicators,shoulder function,and serum inflammatory factors in treating patients with early primary frozen shoulder(PFS).Methods:A total of 168 patients with early PFS admitted to Jiangbei District of The Frist Affiliated Hospital to Army Medical University between June 2021 and June 2023 were selected.They were randomly divided into a control group and an observation group by using a random number table,with 84 patients in each group.The control group received the therapy with MSUS-guided shoulder joint capsule hydraulic expansion.The observation group received the therapy with RI injection on the basis of the control group.After 5 weeks of treatment,the changes of the following parameters were compared and analyzed between the two groups:Visual Analog Scale(VAS)score for shoulder pain,Simple Shoulder Test(SST)score,Constant-Murley Score(CMS),thickness of the glenohumeral joint capsule at the axilla of ultrasound imaging,thickness of coracohumeral ligament(CHL),RI thickness,positivity rate of RI blood flow,passive range of motion(ROM)of the shoulder joint,and serum inflammatory factor levels including tumor necrosis factor-α(TNF-α),C-reactive protein(CRP)and interleukin-6(IL-6).Results:At 5th week after treatment,both groups showed VAS scores decreased,and SST and CMS scores increased.The VAS score(1.34±0.30)of observation group was significantly lower than(1.97±0.46)of control group,and the SST score and CMS score were significantly higher than those of control group(t=10.514,9.597,7.910,P<0.05),respectively.Both groups showed decreases in thickness of the glenohumeral joint capsule at the axilla,CHL thickness,RI thickness,and positivity rate of RI blood flow,and these indicators of observation group were significantly lower than them of control group,with statistically significant differences(t=5.518,16.106,9.050,25.211,P<0.05).The ranges of backward extension and outward rotation of observation group were larger than those of control group,and the differences were statistically significant(t=9.209,12.447,P<0.05).The serum levels of TNF-α,IL-6 and CRP of observation group were significantly lower than those of control group,and the differences were statistically significant(t=10.523,17.750,19.995,P<0.05),respectively.Conclusion:The treatment of MSUS-guided shoulder joint capsule hydraulic expansion combined with RI injection can effectively alleviate the degrees of shoulder pain and dysfunction in patients with early PFS,and improve indicators of ultrasound imaging,the ROM values of outward rotation and backward extension,and reduce serum levels of inflammatory factors,which have favorable therapeutic effects.

Objective This study aims to observe the impact of warm moxibustion on menstrual pain in college students with primary dysmenorrhea and explore its potential mechanism.Methods College students with primary dysmenorrhea were recruited and treated with warm moxibustion at Shenque acupoint for three consecutive menstrual cycles.Healthy subjects were also recruited for comparison.Pain scale,uterine artery hemodynamics,and related inflammatory factors were assessed before and after treatment.Results ①The results of scale study showed that the severity and duration of dysmenorrhea were gradually alleviated with the prolongation of treatment time through the analysis of variance of repeated measurements of the total scores of McGill pain inquiry scale and CMSS dysmenorrhea symptom scale before and after 3 treatments.The results of variance analysis and pairwise comparison of repeated measurements of PRI,VAS and PPI of McGill pain inquiry scale before and after 3 times treatment in warm moxibustion group also showed that each index decreased gradually with the prolongation of treatment time.The comparison of the scores of each item of CMSS scale showed that the severity and duration of low back before treatment were significantly different from those in the healthy group(P<0.001),but the difference was weakened after the third treatment.The severity of vomiting,the duration of vomiting,the severity of diarrhea and the duration of diarrhea were significantly different from those in the healthy group before treatment(P<0.001),but they were still higher than those in the healthy group after the third treatment.but the difference was not statistically significant.②Prior to treatment,PD college students exhibited significantly higher S/D and PI values on both sides compared to healthy subjects,with a statistically significant difference observed for PI on the left side(P<0.001).Following treatment,all aforementioned indexes decreased significantly,particularly PI on the left side which showed a significant difference from pre-treatment levels(P<0.001).③Before treatment,the levels of serum IL-1β,TNF-α,and CRP in PD college students were significantly higher compared to those in the healthy group.The difference in IL-1β level was statistically significant(P<0.001).After treatment,there was a noticeable decrease in the levels of IL-1β,TNF-α,and CRP.Specifically,IL-1β showed a significant reduction(P<0.01),and this time the comparison with the healthy group did not reveal any significant difference in IL-1β levels.Conclusion The application of warm moxibustion at the Shenque acupoint demonstrates a significant improvement in both the dysmenorrhea pain rating index and severity among college students with primary dysmenorrhea,while also alleviating the severity and duration of associated symptoms.These positive effects may be attributed to warm moxibustion's ability to enhance uterine microcirculation in individuals with primary dysmenorrhea,and ameliorating inflammatory conditions.

BACKGROUND:Solute carrier family 1 member 5(SLC1A5)plays a potential role in a variety of diseases,but the exact mechanism of action is unclear.The construction of stable SLC1A5 overexpression and knockdown cell models can provide a powerful experimental tool for in-depth study of the exact role and mechanism of SLC1A5 in diseases and the discovery of potential therapeutic targets. OBJECTIVE:To construct lentiviral vectors for overexpression and knockdown of mouse SLC1A5 and establish stable transfected RAW264.7 cell lines,so as to provide an experimental foundation for further investigation of the role of SLC1A5 in inflammation. METHODS:Primers were designed and synthesized based on the SLC1A5 gene sequence,and the gene segment was amplified using polymerase chain reaction.Subsequently,the target gene segment was directionally inserted into the GV492 vector plasmid,which had been digested with AgeI/NheI enzymes,to construct recombinant lentiviral plasmids.Positive clones were further selected,and their sequences were confirmed.The pHelper1.0 plasmid vector and pHelper2.0 plasmid vector,along with the target plasmid vector,was co-cultured with 293T cells for transfection,resulting in the production and titration of lentiviral stocks.Furthermore,RAW264.7 cells were cultured in vitro,and the working concentration of puromycin was determined.Lentiviruses were separately co-cultured with RAW264.7 cells,and transfection efficiency was determined by measuring fluorescence intensity.Stable transfected cells were selected using puromycin,and real-time fluorescence quantitative PCR and western blot assay were employed to assess the gene and protein expression levels of SLC1A5 in stably transfected cell lines. RESULTS AND CONCLUSION:(1)Sequencing results indicated a perfect match between the sequencing and target sequences,confirming the successful construction of recombinant lentiviral vectors.(2)The titer for the overexpression SLC1A5 lentivirus was 1×109 TU/mL,while the titer for the knockdown SLC1A5 lentivirus was 3×109 TU/mL.(3)The working concentration of puromycin for RAW264.7 cells was determined to be 3 μg/mL.(4)The optimal conditions for transfecting RAW264.7 cells with overexpression/knockdown expression of SLC1A5 lentivirus involved the use of HiTransG P transfection enhancer with a multiplicity of infection value of 50.(5)A significant upregulation of the gene and protein expression levels of SLC1A5 was detected in cell lines stably overexpressing SLC1A5,while gene and protein expression levels of SLC1A5 were significantly decreased in the knockdown stable cell lines.These findings indicate that lentiviral vectors for mouse SLC1A5 overexpression and knockdown have been successfully constructed and a stably transfected RAW264.7 cell line has been obtained.

Objective : To explore the therapeutic effect of extracellular vesicles(EVs) of Aspergillus flavus(A. flavus) on A. flavus infection. Methods: The EVs of A. flavus were isolated using ultracentrifugation and detected/identified by nanoparticle tracking analysis(NTA). Quantitative real-time PCR(qRT-PCR) and enzyme-linked immunosorbent assay(ELISA) kits were used to assess the effect of A. flavus EVs on the polarization of bone marrowderived macrophages(BMDMs) and the expression levels of several cytokines,including tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),interleukin-6(IL-6),and interleukin-10(IL-10). The Galleria mellonella(G.mellonella) infection model was constructed. Three concentration groups of A. flavus EVs(0. 2,2,and 20 μg/larvae) were set as the experimental groups,and PBS was used as the control group for the infection model.Meanwhile,three forms of negative control groups were established,including the PBS group,the pierced controlgroup, and the negative control group. The therapeutic effect of A. flavus EVs on A. flavus infection was evaluated by the survival rate of the G. mellonella infection models. Results: The particle size of A. flavus EVs ranged from 20 to550 nm. A. flavus EVs could polarize BMDMs into both M1 and M2 phenotypes and induce the production of cytokines,including TNF-α,IFN-γ,IL-6,and IL-10. The results of the G. mellonella infection model showed that A.flavus EVs could improve the survival rate of G. mellonella after A. flavus infection. Conclusion The EVs produced by A. flavus can promote the expression of both pro-inflammatory and anti-inflammatory cytokines in BMDMs,induce M1 polarization and M2 polarization of BMDMs,and increase the survival rate of G. mellonella after A. flavus infection.