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.

Circadian rhythms,a result of the long-term evolution of organisms adapting to the Earth′s environment,play a crucial regulatory role in biological activities.Disruptions in circadian rhythms have been iden-tified as potential carcinogenic risk factors,closely linked to the pathological states of digestive system diseases,and significantly contribute to the occurrence and progression of digestive system tumors.These rhythms not only influence the regulation of cell cycles,proliferation,and apoptosis of tumor cells but also profoundly affect their migratory abilities,immune functions,and drug resistance.This article focuses on the specific regulatory mechanisms of circadian rhythm genes in relation to digestive system tumors and comprehensively summarizes research findings on the anti-tumor effects of traditional Chinese medicine targeting these circadian clock genes.The aim is to provide new avenues for target selection and prognostic evaluation in the treatment of digestive system tumors.

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.

OBJECTIVE To investigate the role and mechanisms of the soluble guanylate cyclase(sGC)stimulator sGC003 in improving high altitude pulmonary edema(HAPE)in mice.METHODS Mice were randomly assigned to a normal control group,model group,model+dexamethasone 4 mg·kg-1 group(before modeling,intragastric administration of saline was performed once daily for 6 d,followed by intragastric administration of dexamethasone 4 mg·kg-1 on days 7 and 8),model+riociguat 10 mg·kg-1 group(before modeling,intragastric administration once a day for 7 d),and model+sGC003 5 and 10 mg·kg-1 groups(before modeling,intragastric administration once a day for 7 d).All groups except the normal control group received intratracheal instillation of lipopolysaccharide at a dose of 4 mg·kg-11 h after drug administration on day 7,followed by placement in a hypoxic environment to establish the HAPE model.After 24 h of modeling,the expiratory time,end-inspiratory pause,enhanced pause,and breathing frequency were measured,Lung tissue morphology was examined using HE staining,and lung tissue edema was assessed by determining the wet to dry weight ratio(W/D).The level of interleukin-1β(IL-1β)was determined using immunofluorescence staining.The phosphorylation level of vasodilator-stimulated phosphoprotein(VASP)in lung tissue was analyzed by Western blotting.Additionally,levels of sGC,hypoxia inducible factor-1α(HIF-1α),cyclic guanosine monophosphate(cGMP),IL-6,and IL-1βin serum were quantified using ELISA.RESULTS Compared with the normal control group,the model group had obvious pulmonary edema,and the lung W/D,IL-1β levels,expiratory time,end-inspiratory pause,enhanced pause,as well as serum levels of IL-1β,HIF-1α and IL-6 were significantly increased.Concurrently,the frequency of breathing and serum levels of sGC and cGMP were significantly decreased.Compared with model group,the expiratory time,end-inspiratory pause,enhanced pause,lung W/D and IL-1β levels,and serum levels of IL-1β,HIF-1α and IL-6 were significantly decreased in the model+sGC003 10 mg·kg-1 group;while the frequency of breathing,serum sGC and cGMP levels,phosphorylation level of VASP in lung tissues were significantly increased.CONCLUSION sGC003 can improve lung function,suppress pulmonary inflammation,and mitigate pulmonary edema in HAPE mice by activating the sGC/cGMP pathway.

OBJECTIVE To investigate the role of Bai Ling Long Zao An Shen formula(BLLZ)in sleep improvement in an environmental stress-induced insomnia rat model and explore its underlying mechanisms.METHODS(1)Component analysis:the chemical constituents of the BLLZ extract were analyzed using ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS).(2)Eval-uation of the sedative and hypnotic effect:① Mice:50 ICR mice were randomly divided into normal control group,BLLZ-L group(5,10 and 20 g·kg-1)and diazepam group(DZP,3 mg·kg-1).After five days of intragastric administration,pentobarbital sodium-induced righting reflex and locomotor activity tests were performed.② Rats:8 SD rats were implanted with electrodes and allowed to recover for seven days before baseline EEG data was collected over 24 h.A crossover design(7 d washout period)was employed,with rats randomly assigned to the DZP(3 mg·kg-1)and BLLZ(20 g·kg-1)group.After five days of treatment,24 h EEG recordings were obtained.(3)Insomnia model and interventions:①8 SD rats were allowed to recover for seven days post-surgery,followed by 6 h(14:00-20:00)baseline EEG recording.A 3×3 crossover design was used to assign rats to model(environmental stress-induced insomnia),model+DZP,or model+BLLZ groups.After five days of treatment,insomnia was induced by frequent cage changes(14:00,16:00 and 18:00),and EEG changes were monitored.(4)Mechanistic study:32 SD rats were randomly divided into the normal control group,model group,and model+DZP group.After five days of treatment,hypothalamic tissues were collected for biochemi-cal analysis.γ-aminobutyric acid(GABA),glutamate(Glu),and dopamine(DA)levels were measured using biochemical kits while γ aminobutyric acid receptor subunit alpha-1(GABAA1),core clock proteins period circadian regulator 2(PER2)and circadian locomotor output cycles(CLOCK)protein expressions were assessed by Western blotting.RESULTS(1)Compared with the normal control group,the sleep latency of BLLZ 10 and 20 g·kg-1 and DZP groups was significantly shortened,and the locomotor activity of BLLZ 20 g·kg-1 and DZP groups was significantly reduced;BLLZ 20 g·kg-1 signifi-cantly increased the total sleep time,slow-wave sleep time,and average duration of sleep in normal rats,and significantly reduced the wakefulness time.(2)The total sleep time and slow-wave sleep time of the model group significantly decreased and the wakefulness time significantly increased compared with baseline.(3)Compared with the model group,the total sleep time and slow-wave sleep time of the model+BLLZ group and the model+DZP group were significantly increased,and the wakefulness time significantly shortened.(4)Compared with the normal control group,the Glu/GABA ratio,DA content and CLOCK protein expression were significantly increased and GABAA1 and PER2 protein expres-sion were significantly decreased in the model group;compared with the model group,the Glu/GABA ratio,DA content and CLOCK protein expression were significantly decreased,and the expression of GABAA1 and PER2 were significantly increased in the model+BLLLZ group and the model+DZP group.CONCLUSION BLLZ has sedative and hypnotic effects.It can prolong the total slow-wave sleep time by increasing the average duration of slow-wave sleep episodes,thereby increasing the total sleep time and improving environmental stress-induced insomnia.The mechanism may be related to the downregulation of the Glu/GABA ratio and DA levels as well as the enhancement of GABAA1 expressions and the regulation of hypothalamic core clock protein expressions.

BACKGROUND:With the continuous advancement of medical technology,the combination of oblique lumbar interbody fusion and percutaneous pedicle screw fixation has become a common method for treating lumbar degenerative diseases;however,there is still a lack of in-depth research on the relationship between changes in anterior convexity angles at different postoperative segments and fusion device settlement.OBJECTIVE:To explore the relationship between changes in anterior convexity angle of different segments and Cage subsidence after oblique lumbar interbody fusion combined with percutaneous pedicle screw fixation surgery.METHODS:Patients(93 cases)with lumbar degenerative diseases who underwent oblique lumbar interbody fusion combined with percutaneous pedicle screw fixation surgery in Weifang Hospital of Traditional Chinese Medicine from February 2019 to April 2023 were selected as the research subjects.According to the postoperative disc height loss value,they were divided into Cage subsidence group(25 cases,＞2 mm)and non-Cage subsidence group(68 cases,＜2 mm).Multiple Logistic regression analysis was used to analyze the risk factors for cage subsidence.Stepwise regression was utilized to evaluate the relationship of each risk factor and Cage subsidence.Risk prediction model was built and evaluated.RESULTS AND CONCLUSION:(1)After adjusting for confounding factors,there was still independent correlation between lumbar lordosis loss value and segmental lordosis improvement value and Cage subsidence risk(P＜0.05).(2)Age,oswestry disability index,disc height improvement value,segmental lordosis improvement value,and lumbar lordosis loss value were all independent influencing factors for the occurrence of Cage subsidence(P＜0.05).Among them,age,lumbar lordosis loss value,disc height improvement value,and segmental lordosis improvement value were most closely related to Cage subsidence.(3)The results of multivariate Logistic regression model analysis showed that when P=0.80,the Jordan index was the highest and the prediction effect was the best,with accuracy of 89.27％,sensitivity of 86.67％,and specificity of 89.89％.The model evaluation results showed that it had good discrimination and high accuracy.(4)As the lumbar lordosis loss value and segmental lordosis improvement value increase,the risk of Cage subsidence increases,which affects clinical efficacy.(5)It is indicated that age,lumbar lordosis loss value,disc height improvement value,and segmental lordosis improvement value are most closely related to Cage subsidence,and clinical doctors should pay more attention.