ObjectiveTo explore the role of cuproptosis and identify cuproptosis-related genes in Wilson disease （WD） through bioinformatics analysis and clinical validation，providing implications and directions for the diagnosis and treatment of WD. Methods（1） Screening of target genes： The differentially expressed genes （DEGs） between WD and healthy control were obtained from GeneCards，and the cuproptosis-related genes were obtained from Gene Expression Omnibus （GEO） and published literature.The cuproptosis-related genes in WD were obtained by intersection.Through gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses，the specific biological process，functions or metabolic pathways of cuproptosis-related genes in WD were predicted.Molecular docking and PyMOL visualization were then performed to analyze and verify the potential regulatory mechanism of Gandou Fumu Decoction for cuproptosis.（2）Validation of target genes： The blood samples of 15 WD patients treated in the department of encephalopathy and 15 healthy volunteers undergoing physical examinations in the health management center were randomly collected from the First Affiliated Hospital of Anhui University of Chinese Medicine.The expression levels of target genes were determined by Western blot and real-time PCR. Results（1） A total of 3 607 DEGs in WD were obtained from GSE107323 in GEO，and 68 cuproptosis-related genes were obtained from GeneCards and published literature.Twelve common target genes were obtained by intersection，including three up-related genes（SQSTM1，MIF1，and TAX1BP1） and nine down-regulated genes（CP，SERPINE1，AOC3，GPX4，SLC27A5，VEGF-A，PDHB，PDK1，and ATP7B）.The common target genes were mainly enriched in monocarboxylic acid metabolism，oxidoreductase activity，negative regulation of molecular functions，which mainly involved HIF-1，ferroptosis and other signaling pathways.Molecular docking and PyMOL visualization results showed Gandou Fumu Decoction had good binding ability with the cuproptosis-related genes PDK1，SERPINE1，VEGFA，and AOC3 in WD.（2）A total of 30 blood samples were collected，including 15 WD patients and 15 health volunteers.Western blot results showed that expression levels of target genes were consistent with the results obtained by bioinformatics analysis.RT-qPCR results showed that compared with healthy volunteers，WD patients had down-regulated mRNA levels of SERPINE1，GPX4，SLC27A5，and VEGF-A and up-regulated mRNA levels of SQSTM1 and MIF1（P<0.05）. ConclusionThe expression levels of cuproptosis-related genes in WD patients are consistent with the results predicted by bioinformatics analysis.The characteristic preparation Gandou Fumu Decoction of Xin'an Medicine showed good binding abilities with the cuproptosis-related genes in WD.Cuproptosis may play a key role in the pathophysiological mechanism of WD，which can provide a new target for the diagnosis and treatment of WD.

Objective To discuss the clinical safety,feasibility and efficacy of transcatheter arterial infusion chemotherapy(TAI)combined with lipiodol chemoembolization in the treatment of advanced colorectal cancer(CRC).Methods The clinical data of 37 patients with advanced CRC,who received TAI combined with lipiodol chemoembolization at the First Affiliated Hospital of Zhengzhou University of China between June 2016 and December 2022,were retrospectively analyzed.The clinical efficacy was evaluated,the progression-free survival(PFS)and the serious complications were recorded.Results A total of 55 times of TAI combined with lipiodol chemoembolization procedures were successfully accomplished in the 37 patients.The mean used amount of lipiodol emulsion was 2.9 mL(0.8-10 mL).No serious complications such as bleeding and intestinal perforation occurred.The median follow-up time was 24 months(range of 3-48 months).The postoperative one-month,3-month,6-month and 12-month objective remission rates(ORR)were 67.6%(25/37),67.6%(25/37),64.9%(24/37)and 56.8%(21/37)respectively,and the postoperative one-month,3-month,6-month and 12-month disease control rates(DCR)were 91.9%(34/37),91.9%(34/37),89.2%(33/37)and 81.1%(30/37)respectively.The median PFS was 16 months(range of 2-47 months).As of the last follow-up,22 patients survived and 15 patients died of terminal stage of tumor.Conclusion Preliminary results of this study indicate that TAI combined with lipiodol chemoembolization is clinically safe and effective for advanced CRC,and it provide a new therapeutic method for patients with advanced CRC.

Objective:To investigate the consistency of myocardial extracellular volume between systole and diastole using dual-layer detector spectral CT.Methods:This was a cross-sectional study. Thirty-five patients who underwent cardiac spectral CT examination in West China Hospital of Sichuan University from April 2022 to December 2022 were retrospectively collected. Hematocrit was collected within 3 days before the CT scan. The delayed phases holographic spectral images in systole (45%) and diastole (75%) were obtained using dual-layer spectral CT. CT data were processed using a spectral post-processing workstation, and the extracellular volume (ECV) based on iodine density images, referred as CT-ECV, in systolic and diastolic phases were calculated, respectively. According to the American Heart Association′s 16-segment model of left ventricular, the standard short-axis images were constructed, and the myocardium was standardized into 16 segments at the basal, mid-cavity, and apical levels of the left ventricle. Two radiologists performed a subjective evaluation in the image quality of the CT-ECV images of the whole heart and the three sections in systole and diastole using a "five-point" scale. The ECV of the 16 segments and the whole heart in systole and diastole was calculated. The consistency of subjective evaluations between systole and diastole was assessed using Kappa statistics. Wilcoxon signed-rank tests were used to compare the differences in scores between systole and diastole. Paired sample t-test was used to compare the differences in CT-ECV scores between systole and diastole. The intraclass correlation coefficient was used to test the intra-and inter-observer consistency of CT-ECV measurements between two radiologists. P<0.05 was statistically significant. Results:There was good agreement between the two radiologists on subjective scores of CT-ECV image quality between systole and diastole ( Kappa>0.80), and there was no statistical difference in image quality among the basal, mid-cavity, and apical levels of the left ventricle and whole heart between systole and diastole ( P>0.05). The systolic and diastolic CT-ECV for the entire heart obtained through the delay phase were (33.29±3.46)% and (33.50±3.39)%, respectively, with no statistically significant difference ( t=-0.78, P=0.442). CT-ECV in systole and diastole were (34.15±3.94)% and (35.30±3.99)% for segment 8, (34.03±3.76)% and (35.46±3.74)% for segment 9, and (33.98±3.32)% and (35.05±3.98)% for segment 14, respectively. The mean values of the systolic CT-ECV of segments 8, 9 and 14 were significantly lower than those of diastolic CT-ECV ( t=-2.65, -3.26, -2.42, P=0.012, 0.003, 0.022, respectively). The ICCs for CT-ECV measurements of 16 segments by the two radiologists were greater than 0.90 in both systolic and diastolic, indicating good agreement. Conclusions:There is no significant difference in whole heart CT-ECV values between systolic and diastolic myocardial ECV based on dual-layer spectral CT. However, minor differences (less than 2%) are found between systolic and diastolic myocardial CT-ECV for some segments. Myocardial CT-ECV measurement should be performed on the same segment during the same phase to obtain stable and accurate ECV values.

Objective To investigate the clinical effect of unilateral percutaneous vertebroplasty(PVP)combined with 3D printing technology for the treatment of thoracolumbar osteoporotic compression fracture.Methods A total of 77 patients with thoracolumbar osteoporotic compression fractures from October 2020 to April 2022 were included in the study,all of which were vertebral body compression fractures caused by trauma.According to different treatment methods,they were di-vided into experimental group and control group.Thirty-two patients used 3D printing technology to improve unilateral transpedicle puncture vertebroplasty in the experimental group,there were 5 males and 27 females,aged from 63 to 91 years old with an average of(77.59±8.75)years old.Forty-five patients were treated with traditional bilateral pedicle puncture vertebroplasty,including 7 males and 38 females,aged from 60 to 88 years old with an average of(74.89±7.37)years old.Operation time,intraoperative C-arm X-ray times,anesthetic dosage,bone cement injection amount,bone cement diffusion good and good rate,complications,vertebral height,kyphotic angle(Cobb angle),visual analogue scale(VAS),Oswestry disability index(ODI)and other indicators were recorded before and after surgery,and statistically analyzed.Results All patients were followed up for 6 to 23 months,with preoperative imaging studies,confirmed for thoracolumbar osteoporosis com-pression fractures,two groups of patients with postoperative complications,no special two groups of patients'age,gender,body mass index(BMI),time were injured,the injured vertebral distribution had no statistical difference(P＞0.05),comparable data.Two groups of patients with bone cement injection,bone cement dispersion rate,preoperative and postoperative vertebral body height,protruding after spine angle(Cobb angle),VAS,ODI had no statistical difference(P＞0.05).The operative time,intra-operative fluoroscopy times and anesthetic dosage were statistically different between the two groups(P＜0.05).Compared with the traditional bilateral puncture group,the modified unilateral puncture group combined with 3D printing technology had shorter operation time,fewer intraoperative fluoroscopy times and less anesthetic dosage.The height of anterior vertebral edge,kyphosis angle(Cobb angle),VAS score and ODI of the affected vertebrae were statistically different between two groups at each time point after surgery(P＜0.05).Conclusion In the treatment of thoracolumbar osteoporotic compression fractures,3D printing technology is used to improve unilateral puncture PVP,which is convenient and simple,less trauma,short operation time,fewer fluoroscopy times,satisfactory distribution of bone cement,vertebral height recovery and kyphotic Angle correction,and good functional improvement.

Objective This study aimed to assess the clinical efficacy of Shenling Baizhu Granules in treating low anterior resection syndrome(LARS)in rectal cancer.Methods The study employed a randomized,double-blind,placebo-parallel controlled,single-center,validity-tested clinical trial design.December 2019 to June 2022,the Department of Gastrointestinal Surgery and Integrated Traditional Chinese and Western Medicine of Peking University First Hospital recruited 110 patients who had undergone low anterior resection(LAR)for rectal cancer and subsequently developed LARS.These patients,meeting the enrollment criteria,were randomly assigned into the treatment group(55)and the control group(55)using the double-blind method principle.The randomization table was generated by SAS 9.2 software employing the double-blind method.The treatment group received oral Shenling Baizhu Granules,while the control group received oral placebo granules.Both groups commenced treatment on the 10th day after-surgery for 30 consecutive days.Patients were evaluated using LARS score,traditional Chinese medicine(TCM)symptom grading,and XU Zhongfa score before treatment,on the 15th day of treatment,and on the 1st day after treatment cessation.Results Out of 110 patients,107 were included in the full analysis set for efficacy analysis:55 patients in the treatment group and 55 patients in the control group.One case in the treatment group was excluded(against protocol),and two cases in the control group were excluded(one lost to follow-up,one against protocol).Baseline data between the two groups were consistent,with no statistically significant difference.Before treatment,LARS scores for the treatment and control groups were 33.0(31.0,36.0)and 34.0(32.0,37.0)respectively.Patients with TCM symptom scores of grades 2 to 3 accounted for 92.73％and 90.57％in the treatment and control groups,respectively,with no statistically significant difference.After 30 days of treatment,LARS scores for the treatment and control groups were 21.0(19.8,23.0)and 26.0(22.0,28.0)respectively.The percentage of patients with TCM symptom scores of grades 2 to 3 decreased to 33.33％in the treatment group and 66.04％in the control group,with a statistically significant difference.Shenling Baizhu Granules showed rapid improvement in watery or loose stools in post-operative rectal cancer patients.After 30 days of treatment,Shenling Baizhu Granules significantly improved appetite,stool consistency,abdominal distension,abdominal pain,and eructation symptoms in postoperative rectal cancer patients.Before treatment,the XU Zhongfa scores for the treatment and control groups were 3.0(2.0,4.3)and 4.0(2.0,4.0)respectively,with no statistically significant difference.After 30 days of treatment,the XU Zhongfa scores for the treatment and control groups were 7.0(6.0,8.0)and 6.0(5.0,7.0)respectively,with the treatment group significantly higher than the control group(P＜0.01).Conclusion Shenling Baizhu Granules can effectively improve LARS symptoms in patients following LAR of rectal cancer within a short period of time.

Objective:To explore the effects of hesperidin against human tongue squamous cell carcinoma Tca-8113 cells and explore its mechanism of action.Methods:Tca-8113 cells were cultured in vitro with different concentrations of hesperidin.MTT assay and flow cytometry were used to detect the cell proliferation and apoptosis respectively.The potential targets of hesperidin against Tca-8113 cells were studied by network pharmacology.The protein-protein interaction network(PPI)was constructed,gene ontology GO analy-sis,kyoto gene and genome baike encyclopedia(KEGG)enrichment analysis were performed.Q-PCR was used to detect the effect of different concentrations of hesperidin on the related mRNA expression level of Tca-8113 cells.The binding force of hesperidin to the core targets was predicted by molecular docking and a scoring function was used to judge the binding force.Results:Hesperidin inhibi-ted proliferation and promoted apoptosis of Tca-8113 cells(P＜0.05)in a dosedependant manner;KEGG enrichment analysis of the key gene targets involved in the regulation of oral squamous cell carcinoma by hesperidin may be related to PI3K-Akt,MAPK and Ras signaling pathways.Q-PCR results showed that the mRNA expression of MAPK8,HSP90AA1 and MAPK1(ERK2)were significantly different(P＜0.05).Molecular docking results showed that hesperidin had the strongest binding force with MAPK1(ERK2).Conclu-sion:Hesperidin may inhibit the proliferation and promote apoptosis of Tca-8113 cells by regulating Ras/Raf/MEK/ERK signaling pathway.

Objective To investigate the effect of pristimerin(PM)on adriamycin(ADM)resistance in osteosarcoma cells and its mechanism.Methods The ADM resistant cells MG-63/ADR was established in vitro by using the ADM increasing concentration gradient long-term culture method,CCK-8 was used to detect the effects of different concentrations of ADM on the viability of osteosarcoma cells MG-63 and MG-63/ADR cells to verify the drug resistance of ADM;subsequently,MG-63/ADR cells were randomly divided into the MG-63/ADR group(normal culture),low-dose PM group(adding 0.5 μmol/L PM),middle-dose PM group(adding 1.0 μmol/L PM)and high-dose PM group(adding 2.0 μmol/L PM).The cell viability of each group under different concentrations of ADM treatment was detected by CCK-8;the sensitivity of cells to ADM in each group was detected by plate clone formation assay;the cell morphology of each group was observed under microscope;the cell apoptosis level was detected by flow cytometry;Western blot was used to detect the expression of phosphorylated mammalian STE20-like protein kinase 1(p-MST1),MST1,phosphorylated large tumor suppressor 1(p-LATS1),LATS1,phosphorylated Yes associated protein(p-YAP)and YAP proteins.Results With the intervention of 25,125,625,3 125 and 15 625 ng/mL ADM,the activity of MG-63/ADR cells was significantly higher than that of MG-63 cells(P＜0.05),which indicated that the ADM resistant cell model was successfully established.Compared with the MG-63/ADR group,the number of cells in the low,middle and high-dose PM groups decreased,and the cells gradually shrank,the spacing between cells became larger,and the cell viability,cell clonal formation rate and p-YAP/YAP level decreased(P＜0.05),the apoptosis rate and the levels of p-MST1/MST1 and p-LATS1/LATS1 significantly increased(P＜0.05).Conclusion The inhibitory effect of PM on ADM resistance in osteosarcoma cells may be related to the activation of Hippo pathway and the downregulation of YAP signal.

Objective:To investigate the effect of CD226 gene knockout (CD226 -/-) on carbon tetrachloride (CCl 4) -induced liver fibrosis in mice. Methods:Eight 6-8 week-old male wild-type and CD226 -/- C57BL/6 mice were used, with weights of (22.16±2.24) g and (21.84±2.50) g, respectively. Four mice each from wild-type and CD226 -/- mice were injected intraperitoneally with CCl 4 to establish a liver fibrosis model, namely, the liver fibrosis group and the CD226 -/- liver fibrosis group, and the remaining four mice each corresponded to the control group and the CD226 -/- control group. The effect of CD226 -/- on liver mass, liver index and Ishak fibrosis score were observed. HE, Masson, and Sirius red staining were performed to observe the liver injury and liver fibrosis. Immunohistochemistry and Western blot were used to detect the protein expression levels of collagen type I (Collagen I) and α-smooth muscle actin (α-SMA) in liver tissues. Flow cytometry was used to analyze the proportion of T lymphocytes and their intracellular IFN-γ secretion levels in the peripheral blood and liver tissues. The mRNA expression level of transforming growth factor-β1 (TGF-β1) in the liver tissues was detected by real-time fluorescence quantitative PCR (qPCR). Results:Compared to the liver fibrosis group, the CD226 -/- liver fibrosis group showed an increase in liver mass [(2.00±0.14) g vs. (1.41±0.16) g] and liver index (7.25±0.59 vs. 5.33±0.30), and a decrease in Ishak's fibrosis score [(6.75±0.96) score vs. (10.00±1.41) score], and the differences were statistically significant (all P<0.05). HE staining showed disorganization of the liver tissue structure, and degeneration and necrosis of the hepatocytes both occurred in the liver fibrosis group and CD226 -/- liver fibrosis group, but the degree of inflammatory cell recruitment was milder in the CD226 -/- liver fibrosis group than the liver fibrosis group. Sirius red and Masson staining revealed reduced collagen fibre deposition in the CD226 -/- liver fibrosis group compared to the liver fibrosis group. Immunohistochemical staining showed that the expression of Collagen I [(4.38±0.51)% vs. (6.55±1.40)%] and α-SMA [(0.77±0.20)% vs. (1.20±0.24)%] were reduced in the CD226 -/- liver fibrosis group compared to the liver fibrosis group, and the differences were statistically significant (both P<0.05). Western blot indicated that the relative expression level of Collagen I was reduced in the CD226 -/- liver fibrosis group compared to the liver fibrosis group (0.35±0.14 vs. 1.66±0.73), and the difference was statistically significant ( P<0.05). Flow cytometry revealed increased intracellular IFN-γ secretion by T lymphocytes in the peripheral blood in the CD226 -/- liver fibrosis group compared to the liver fibrosis group. qPCR result revealed that the relative mRNA expressions of TGF-β1 was reduced in the CD226 -/- liver fibrosis group compared to the liver fibrosis group (0.38±0.18 vs. 0.61±0.28), and the difference was statistically significant ( P<0.05). Conclusion:CD226 -/- attenuates CCl 4-induced hepatic inflammatory infiltration and hepatic fibrosis, which may be related to its affected intrahepatic T lymphocytes and intracellular IFN-γ secretion.

Objective:To analyze the current prevalence of pre-diabetes (PDM) and its relationship with overweight and obesity in an adult health check-up population.Methods:This study was a cross-sectional and retrospective cohort study and was applied using whole-cluster random sampling method. A total of 491 379 adults who underwent health check-ups at the Health Management Centre of Sichuan Provincial People′s Hospital from January 2017 to July 2023 were selected to analyze the epidemiological characteristics of PDM and overweight-obesity, as well as the trend of change over time. A retrospective cohort study was conducted on 19 001 of the subjects who underwent≥3 health check-ups and did not have diabetes and PDM at baseline, and the relationships between body mass index, waist circumference and the risk for developing PDM were analyzed using Cox proportional risk regression models. And the dose-response relationship between body mass index, waist circumference and the risk for developing PDM was analyzed using restricted cubic spline regression (RCS).Results:Of the 491 379 cases included in the cross-sectional study, 275 084 were male and 216 295 were female, 163 158 cases were under 40 years old, and 328 221 cases were 40 years old and above; the total prevalence of PDM was 19.41% in 2017-2023, with an overall increasing trend. Of the 19 001 people included in the cohort study, a total of 2 487 (13.09%) new cases of PDM were identified at the end of follow-up. After adjusting for confounding factors, overweight ( HR=1.150, 95% CI: 1.047-1.263), obesity ( HR=1.335, 95% CI: 1.149-1.552) and abdominal obesity ( HR=1.218, 95% CI: 1.105-1.342) were risk factors for PDM. The risk of PDM rised with the increase of body mass index (>22.9 kg/m 2, Pnon-linear=0.973) and waist circumference (>80 cm, Pnon-linear=0.830), with a linear dose-response mode. In different gender and age groups, it was found the greater the body mass index (>24.1 kg/m 2 for men,>21.5 kg/m 2 for women;>23.3 kg/m 2 for age≥40 years,>24.1 kg/m 2 for age<40 years) and waist circumference (>85 cm for men, >73 cm for women; >82 cm for age ≥40 years, >85 cm for age <40 years), the higher the risk of PDM. Conclusions:The prevalence of PDM is on the rise in the adult health check-up population. To prevent PDM, it is necessary to control the body mass index and waist circumference to a lower level than the overweight and obesity standards.

The formation of learning and memory is regulated by synaptic plasticity in hippocampal neurons. Here we explored how gestational exposure to dexamethasone, a synthetic glucocorticoid commonly used in clinical practice, has lasting effects on offspring's learning and memory. Adult offspring rats of prenatal dexamethasone exposure (PDE) displayed significant impairments in novelty recognition and spatial learning memory, with some phenotypes maintained transgenerationally. PDE impaired synaptic transmission of hippocampal excitatory neurons in offspring of F1 to F3 generations, and abnormalities of neurotransmitters and receptors would impair synaptic plasticity and lead to impaired learning and memory, but these changes failed to carry over to offspring of F5 and F7 generations. Mechanistically, altered hippocampal miR-133a-3p-SIRT1-CDK5-NR2B signaling axis in PDE multigeneration caused inhibition of excitatory synaptic transmission, which might be related to oocyte-specific high expression and transmission of miR-133a-3p. Together, PDE affects hippocampal excitatory synaptic transmission, with lasting consequences across generations, and CDK5 in offspring's peripheral blood might be used as an early-warning marker for fetal-originated learning and memory impairment.