ObjectiveTo explore the mechanism by which exosomes derived from gastric cancer cells MGC-803 promote tumor progression through the upregulation of ABCG8 expression. MethodsUltra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） lipidomics， quantitative real-time PCR （qPCR） and Western blot were used to verify the role of ABCG8 in gastric cancer progression. Exosomes derived from gastric cancer cells were introduced， and a series of in vitro experiments were conducted to evaluate their effect on ABCG8 expression， as well as on cell proliferation， migration， and clonogenic ability in normal gastric mucosal epithelial GES-1 cells. ResultsUPLC-MS/MS lipidomics analysis revealed significant changes in triacylglycerol （TG） among the differential lipids. qPCR and Western blot analysis showed that the mRNA and protein expression levels of ABCG8 in the GC-Exo-treated group were higher than those in the GES-1 cell group （P<0.05）. Colony formation and Transwell migration assays revealed that， compared with the GES-1 cell group， the GC-Exo-treated group exhibited a greater number of colonies （P<0.05） and migrated cells （P<0.05） after 48 h of treatment. CCK-8 assay results showed that the proliferation rate in the GC-Exo-treated group （50 μg/mL） was higher than that in the GES-1 cell group （P<0.001）. Lipidomics analysis revealed that， compared with the GES-1 cell group， TG levels were upregulated in the GC-Exo-treated group， accompanied by decreased levels of phosphatidylcholine （PC） PC（38：2e）. ConclusionExosomes derived from gastric cancer cells MGC-803 may promote tumor progression by upregulating ABCG8 expression to mediate cholesterol efflux， thereby triggering pro-carcinogenic lipid metabolic reprogramming in normal gastric mucosal epithelial cells.

Objective:To investigate the reliability and validity of different measurement methods under 30° arthroscopy for quantifying the proportion of glenoid bone defect in bony Bankart lesions of the shoulder joint and validate its preliminary application effect.Methods:Eight intact shoulder glenoid specimens were selected, with no existing defects or deformities, from donors of 4 females and 4 males, with their age at death of 43-67 years [(54.4±8.0)years]. Bone defects of 12.5% and 25% were created in the glenoid at 0° and 45° relative to the longitudinal axis, with two specimens per defect category. The defect proportion in each specimen was quantified using direct measurement and CT-based digital reconstruction and these values served as reference standards for subsequent statistical analysis. Using a combined approach of arthroscopic simulation equipment and cadaveric study, five investigators performed simulated examinations through the standard posterior portal (2 cm medial and 1.5 cm inferior to the posterolateral acromial corner) and the modified posteroinferior portal (2 cm medial and 3 cm inferior to the posterolateral acromial corner) separately. Under 30° arthroscopy, the glenoid bone loss percentage was measured using the bare spot method and secant chord method. The reliability was analyzed for these measurements. Furthermore, using direct physical measurements and CT-based three-dimensional reconstruction data from the same specimens as reference standards, the comprehensive validity of four measurement methods was evaluated (standard posterior portal-bare spot method, standard posterior portal-secant chord method, modified posteroinferior portal-bare spot method, and modified posteroinferior portal-secant chord method). The independent validity of each method was assessed according to bone defect morphology classification to determine differences in measurement accuracy across defect types. In an arthroscopic procedure for a patient with Bigliani type IIIB bony Bankart lesion, the standard posterior portal-secant chord method was applied to quantify the proportion of glenoid bone defects.Results:The mean reference values from direct measurement and CT measurement of glenoid bone defect proportion in eight bony Bankart lesion specimens were 12.71%/12.37%, 13.17%/13.10%, 25.71%/24.9%, 26.6%/26.95%, 13.41%/13.10%, 12.90%/12.59%, 26.42%/25.94%, and 26.73%/27.06%, respectively. Measurements obtained by the five investigators showed intraclass correlation coefficients (ICCs) all greater than 0.90, indicating excellent interobserver agreement. In the validity analysis, the standard posterior portal-secant chord method demonstrated the highest overall validity. Using direct measurement and CT-based measurement as reference standards, the overall validity was (0.90±0.38)% and (1.07±0.53)% for the standard posterior portal-bare spot method, (1.33±0.40)% and (1.51±0.54)% for the modified posteroinferior portal-bare spot method, and (0.53±0.17)% and (0.70±0.38)% ( P<0.05) for the modified posteroinferior portal-secant chord method. In contrast, the standard posterior portal-secant chord method showed an overall validity of (0.10±0.10)% and (0.28±0.39)% ( P>0.05). In subsequent independent validity analyses, the standard posterior portal-secant chord method also demonstrated superior validity across all bone defect subtypes over the other three methods. In a patient with a Bigliani type IIIB bony Bankart lesion, we used the standard posterior portal-secant chord method to quantify the glenoid bone loss in 2 minutes, revealing a defect proportion of 26.6%. An arthroscopic autologous iliac bone graft procedure with single-tunnel elastic fixation guided by this measurement achieved favorable outcomes, with stable reduction, secure internal fixation and favorable recovery of shoulder function at 2 months postoperatively. Conclusion:For various types of bony Bankart lesions, the 30° arthroscopic standard posterior portal-secant chord method provides the most accurate quantification of glenoid bone loss and its preliminary clinical application yields satisfactory results.

Objective To analyze the expression and clinical significance of stimulator of interfer-on genes(STING),C-C motif chemokine ligand 5(CCL5),interferon regulatory factor 3(IRF3)and programmed death ligand-1(PDL1)in squamous cell lung cancer.Methods A total of 56 pa-tients with squamous cell lung cancer were enrolled.Resected tumor tissues and adjacent non-tumor tissues(located more than 5 cm from the tumor margin)were collected.Immunohistochemical staining was performed to detect the expression of STING,CCL5,IRF3 and PDL1.The correlations of STING,CCL5,IRF3 and PDL1 with clinical data were analyzed.The relationship between the expression of STING,CCL5,IRF3 and PDL1 in lung squamous cell carcinoma tissues and prognosis was also evalua-ted.The prognostic factors of patients with lung squamous cell carcinoma were analyzed.Results The positive rate of STING expression in lung squamous cell carcinoma tissues was significantly lower than that in adjacent non-tumor tissues,whereas the positive rates of CCL5,IRF3 and PDL1 were significantly higher(P＜0.05).The expression levels of STING,CCL5,IRF3 and PDL1 were associated with tumor diameter,TNM stage,lymph node metastasis and differentiation degree(P＜0.05).The 3-year survival rate of STING positive expression patients was significantly higher than that of STING negative expression patients(P＜0.05).The 3-year survival rate of CCL5 positive,IRF3 positive and PDL1 positive expression patients was significantly lower than that of CCL5 negative,IRF3 negative and PDL1 negative expression patients(P＜0.05).STING,CCL5,IRF3 and PDL1 were identified as prognostic factors for patients with squamous cell lung cancer(P＜0.05).Conclusion In squamous cell lung cancer tissues,STING is expressed at low levels,while CCL5,IRF3 and PDL1 are ex-pressed at high levels.These findings have significant clinical value in assessing the prognosis of pa-tients with squamous cell lung cancer.

Objective To develop an ultra high performance liquid chromatography-mass spectrometry(UPLC-MS)method for quantifying serum levels of norepinephrine(NE)and 5-hydroxytryptamine(5-HT),and to monitor the dynamic changes in these neurotransmitters during the process of establishing a model of acute reserpine-induced depression-like mice.Methods By evaluating matrix effect,recovery,precision,and accuracy efficiency,an UPLC-MS method for determining the concentrations of NE and 5-HT in serum was established.Forty-eight C57 mice were randomly divided into normal control and model groups,which were intraperitoneally injected with physiological saline(10 mL/kg)and reserpine(2.5 mg/kg),respectively.At various time points after intraperitoneal injection,the degree of ptosis and decreases in body temperature of the mice were observed before orbital blood was sample for the determination of NE and 5-HT levels.Results The concentrations of NE and 5-HT showed good linearity within the range of 15.63 to 2000.00 ng/mL,with R2 values greater than 0.999.The results of methodological validation met the requirements for the analysis of biological samples,with a lower limit of quantification of 15.63 ng/mg.After intraperitoneal injection of reserpine,the model mice exhibited varying body temperature decreases and ptosis.At 1 and 2 h post-administration,the depression-like symptoms in the model group were significantly different from those of the normal control group(P＜0.01).The body temperature of mice in the model group was significantly lower than that of mice in the normal control group(P＜0.01),while the score of the eyelid ptosis was significantly higher(P＜0.001).The levels of NE and 5-HT in the serum of model mice were also significantly depleted,and were significantly different from those of the normal control group at 0.5,1 and 2 h(P＜0.05).Conclusion The study process of established a rapid and accurate method for dynamically observing the changes in NE and 5-HT levels during the process of establishing a model of acute reserpine-induced depression-like mice,which might contribute to the study of the pathogenesis of depression and the development of new antidepressant drugs.

Objective To investigate the pharmacological mechanism through which total flavonoids extracted from Xiaobuxin-Tang(XBXT-2)affects neural network activities in the frontal cortex by focusing on the effects of XBXT-2 on the cortical field potentials in the frontal association cortex(FrA)in mice.Methods Cortical electrodes were implanted into the skull of C57BL/6J mice targeting the FrA.After a 7-day recovery period,the mice were administered XBXT-2 intragastrically at a dose of 100 mg/kg,and 1 hour later,local field potential(LFP)in the FrA were recorded for 30 minutes.Spectral analysis of the data was performed using Neuro Explorer software.Changes in the power spectral density of α,β,θ,γ,and δ frequency bands before and after drug administration were analyzed using GraphPad Prism 10.3.Phase-amplitude coupling of θ and γ oscillations was analyzed using Matlab 2021 software.Results It was found that the oral administration of XBXT-2 significantly suppressed high-frequency γ oscillations while simultaneously enhancing θ,β,α,and δ oscillations in FrA of mice compared to the control.Furthermore,XBXT-2 treatment markedly strengthened the phase-amplitude coupling between θ and γ oscillations.Conclusion XBXT-2 possibly affects emotional and cognitive functions by modulating neural network activity in FrA and enhancing θ-γ phase-amplitude coupling in mice.

OBJECTIVES: To investigate the protective effects of graphene heating film far-infrared (FIR) hyperthermia therapy against frostbite in mice and its impacts on microcirculation and coagulation function. METHODS: Seventy-six C57BL/6J mice were randomized into control, model, graphene-FIR, and carbon fiber-FIR groups. After 7-day FIR intervention (4 h/day), the mice were subjected to acute (4 ℃, 4 h) and intermittent (4 ℃, 4 h/day for 3 days) cold exposure and the changes in rectal temperature were monitored. In liquid nitrogen frostbite experiment, 24 ICR mice were divided into model, graphene-FIR, and carbon fiber-FIR groups, and after a 7-day FIR pretreatment (4 h/day), the liquid nitrogen frostbite models were established and apparent scores of the wounds were assessed on days 3 and 6 after modeling. In carrageenan-induced thrombosis experiment, 40 ICR mice were allocated to control, model, graphene-FIR, carbon fiber-FIR, and prazosin groups to test the effect of a 7-day FIR intervention on thrombosis induced by intraperitoneal carrageenan injection (2.5 mg/kg) by measuring thrombus length, blood perfusion, and serum biomarkers (6-keto-PGF1α, TXB2, t-PA, IL-6, IL-1β, TNF‑α) 24 h after the injection. RESULTS: The mice in graphene-FIR group showed significantly elevated rectal temperature in cold exposure tests. In mice with liquid nitrogen-induced frostbite, graphene-FIR treatment significantly reduced the wound scores and reduced frostbite area, producing better effects than carbon fiber. In mice with carrageenan-induced thrombosis, graphene-FIR treatment significantly decreased tail thrombosis length and thrombosis area, increased blood perfusion, lowered serum levels of TXB2, TNF-α and IL-6, and increased the levels of 6-keto-PGF1α and t-PA. CONCLUSIONS Graphene heating film FIR therapy can alleviate frostbite injury in mice by improving microcirculation, suppressing thrombosis and inflammatory responses, and reducing coagulation dysfunction.
Animals ; Frostbite/therapy* ; Graphite ; Mice, Inbred C57BL ; Mice ; Infrared Rays ; Mice, Inbred ICR ; Hyperthermia, Induced/methods* ; Male ; Microcirculation

OBJECTIVES: To assess the regulatory effect of cannabidiol (CBD) on circadian rhythm sleep disorders following general anesthesia and explore its potential mechanism in a rat model of propofol-induced rhythm sleep disorder. METHODS: An electrode was embedded in the skull for cortical EEG recording in 24 male SD rats, which were randomized into control, propofol, CBD treatment, and diazepam treatment groups (n=6). Eight days later, a single dose of propofol (10 mg/kg) was injected via the tail vein with anesthesia maintenance for 3 h in the latter 3 groups, and daily treatment with saline, CBD or diazepam was administered via gavage; the control rats received only saline injection. A wireless system was used for collecting EEG, EMG, and body temperature data within 72 h after propofol injection. After data collection, blood samples and hypothalamic tissue samples were collected for determining serum levels of oxidative stress markers and hypothalamic expressions of the key clock proteins. RESULTS: Compared with the control rats, the rats with CBD treatment showed significantly increased sleep time at night (20:00-6:00), especially during the time period of 4:00-6:00 am. Compared with the rats in propofol group, which had prolonged SWS time and increased sleep episodes during 18:00-24:00 and sleep-wake transitions, the CBD-treated rats exhibited a significant reduction of SWS time and fewer SWS-to-active-awake transitions with increased SWS aspects and sleep-wake transitions at night (24:00-08:00). Diazepam treatment produced similar effect to CBD but with a weaker effect on sleep-wake transitions. Propofol caused significant changes in protein expressions and redox state, which were effectively reversed by CBD treatment. CONCLUSIONS CBD can improve sleep structure and circadian rhythm in rats with propofol-induced sleep disorder possibly by regulating hypothalamic expressions of the key circadian clock proteins, suggesting a new treatment option for perioperative sleep disorders.
Animals ; Rats, Sprague-Dawley ; Male ; Cannabidiol/therapeutic use* ; Rats ; Circadian Rhythm/drug effects* ; Propofol/adverse effects* ; Anesthesia, General/adverse effects* ; Sleep Wake Disorders/chemically induced* ; Hypothalamus/metabolism* ; Electroencephalography

OBJECTIVE: To observe the clinical effect of acupuncture and moxibustion combined with umbilical therapy on anxiety and depression in patients with neurogenic bladder (NB) after spinal cord injury (SCI). METHODS: Thirty cases of NB after SCI with anxiety and depression were selected and treated with acupuncture and moxibustion combined with umbilical therapy. Acupuncture was applied at Baihui (GV20), Yintang (GV24⁺), Sanyinjiao (SP6), Shenmen (HT7), Hegu (LI4), Taichong (LR3), once a day, continuous treatment for 4 weeks. Ginger moxibustion was applied at the bladder meridian of foot taiyang and governor vessel, once a day, continuous treatment for 4 weeks. In treatment of umbilical therapy, Chaihu (Radix Bupleuri), Yujin (Radix Curcumae), Rougui (Cortex Cinnamomi) were ground and mixed with the same amount of honey, put into the application, and the application was placed on the navel after filling the navel with fine salt, once a day for 4 weeks. Hamilton anxiety scale (HAMA) score, Hamilton depression scale (HAMD) score, urodynamic indexes (maximum urinary flow rate [Qmax], maximum detrusor pressure [Pdet-max], residual urine volume [RUV]), neurogenic bladder symptom score (NBSS), urinary symptom distress scale (USDS) score were compared before and after treatment, and the clinical efficacy was evaluated. RESULTS: After treatment, the scores of HAMA, HAMD, NBSS, USDS and RUVwere lower than those before treatment (P<0.05), and Qmax and Pdet-max were higher than those before treatment (P<0.05). The total effective rate was 93.3 (28/30). CONCLUSION Acupuncture and moxibustion combined with umbilical therapy can effectively relieve anxiety and depression symptoms, improve urination disorders in patients with NB after SCI.
Humans ; Moxibustion ; Male ; Female ; Adult ; Middle Aged ; Acupuncture Therapy ; Spinal Cord Injuries/psychology* ; Depression/etiology* ; Anxiety/etiology* ; Urinary Bladder, Neurogenic/etiology* ; Young Adult ; Aged ; Combined Modality Therapy ; Acupuncture Points

Objective To explore the effect of acupuncture combined with manual reduction on pain and lumbar function in elderly patients with degenerative lumbar spondylolisthesis under the guidance of the Bone Strengthening and Tendon Softening Theory.Methods Ninety elderly patients with lumbar spondylolisthesis treated in our hospital from January 2021 to April 2023 were selected as study subjects.Based on a random number table method,the participants were divided into experimental group and control group,each consisting of 45 cases.The control group received acupuncture treatment,while the experimental group received manual reduction treatment on top of that.Both groups underwent continuous treatment for 4 weeks.We compared the two groups in Traditional Chinese Medicine(TCM)syndrome score,Visual Analog Scale(VAS)score,Japanese Orthopaedic Association(JOA)lumbar function score,Oswestry Disability Index(ODI),serum inflammatory factor level,and clinical efficacy between before and after treatment.Results Before treatment,there was no statistically significant difference in TCM syndrome scores or VAS scores between the two groups(P＞0.05).After treatment,both TCM syndrome scores and VAS scores significantly decreased in both groups,with the experimental group showing significantly lower scores than the control group(P＜0.05).Before treatment,there was no statistically significant difference in JOA scores and ODI scores between the two groups(P＞0.05).After treatment,JOA scores significantly increased in both groups,with the experimental group showing significantly higher scores than the control group(P＜0.05).ODI scores significantly decreased in both groups,with the experimental group showing significantly lower scores than the control group(P＜0.05).Before treatment,the two groups did not differ significantly in serum IL-6,TNF-α,or CRP levels(P＞0.05).After treatment,serum IL-6,TNF-α,and CRP levels significantly decreased in both groups,with the experimental group showing lower levels than the control group(P＜0.05).The total effective rate in the experimental group was 88.89％,significantly higher than 68.89％ in the control group(P＜0.05).Conclusion Acupuncture combined with manual reduction guided by the Bone Straightening and Tendon Softening theory can effectively alleviate pain,improve various TCM syndromes,enhance lumbar function level,and show significant clinical efficacy in patients.