Drug-induced liver injury （DILI） is the main cause of failures in drug development and the withdrawal of approved drugs from the market， and therefore， there is an increasing demand for accurate prediction and in vitro testing. However， the two-dimensional cell culture system of hepatocytes is not suitable for the toxicity study of long-term drug use due to the fact that it cannot accurately simulate and reproduce the real environment and micro-ecosystem of hepatocytes in vivo. In view of this， there is an urgent need for liver models with higher predictability to assess the hepatotoxicity of drugs in drug development and the safety evaluation of active compounds. This article reviews the construction and application of three-dimensional in vitro hepatocyte culture systems for DILI， in order to provide a reference for their effective implementation in DILI analysis.

Objective: To investigate the value of decreased carbohydrate antigen 19-9(CA19-9) kinetics in predicting short-term outcomes and determining prognosis among advanced biliary or pancreatic cancer patients receiving first-or second-line therapy in the real world. Methods : Eighty-nine patients were retrospectively collected with advanced biliary or pancreatic cancer, especially on the CA19-9 dynamics and decline rates at different time points. This study evaluated the association of CA19-9 changes with clinicopathological features, short-term response to antitumor therapy, and survival outcomes. Results : The enrolled patients recorded baseline CA19-9 levels ranging from 1.20 to 65 706.40 U/ml, with a median of 303.11 U/ml. There was no statistical correlation between baseline CA19-9 levels and gender, age, body mass index, primary tumor site, hepatic metastases, pulmonary metastases, lymph node metastases, peritoneal metastases, performance status, treatment lines, and combinations of drug types. Baseline CA19-9 levels were not associated with systemic immunoinflammatory index, prognostic nutritional index, and total bilirubin. A 25% or 50% decrease in CA19-9 after 2-3 therapy courses indicated short-term efficacy in reaching tumor objective remission or disease control. Both combinations of multiple drug types and a 25% decline in CA19-9 after one course of treatment were independent prognostic factors that affected the longer progression-free survival of patients receiving first or second line of treatment. Conclusion Decreased CA19-9 kinetics has specific values in predicting the efficacy and prognosis of advanced biliary or pancreatic cancer.

Mastitis,as an important disease that endangers the development of the dairy industry,causes huge economic losses to the breeding industry.Staphylococcus aureus is one of the impor-tant pathogenic bacteria that cause mastitis in dairy cows.Previous studies have found that deoxy-cholic acid(DCA)plays an important role in many diseases,but its effect on mastitis has not been reported yet.In order to investigate the effect and mechanism of DCA on Staphylococcus aureus in-duced mouse mastitis,this study constructed a Staphylococcus aureus induced mouse mastitis model,mice were intraperitoneally injected DCA(0.01,0.02,0.03 mg/g),breast tissue were col-lected,and the protective effect and mechanism of DCA on mastitis were evalued.The results showed that DCA dose dependently reduced the pathological damage of mouse breast tissue in-duced by Staphylococcus aureus;DCA inhibited the expression of inflammatory cytokines TNF-α and IL-1β and MPO activity induced by Staphylococcus aureus;DCA significantly reduced the con-centration of MDA and iron ions,while upregulating the expression of GSH and GPX4,indicating that DCA could inhibit ferroptosis induced by Staphylococcus aureus;Western blot results showed that DCA could upregulate TGR5 receptors,thereby inhibiting the NF-κB signaling pathway.The above results indicate that DCA inhibits mastitis by regulating the TGR5-NF-κB signaling pathway to inhibit the inflammatory response and ferroptosis induced by Staphylococcus aureus.

Mastitis,as an important disease that endangers the development of the dairy industry,causes huge economic losses to the breeding industry.Staphylococcus aureus is one of the impor-tant pathogenic bacteria that cause mastitis in dairy cows.Previous studies have found that deoxy-cholic acid(DCA)plays an important role in many diseases,but its effect on mastitis has not been reported yet.In order to investigate the effect and mechanism of DCA on Staphylococcus aureus in-duced mouse mastitis,this study constructed a Staphylococcus aureus induced mouse mastitis model,mice were intraperitoneally injected DCA(0.01,0.02,0.03 mg/g),breast tissue were col-lected,and the protective effect and mechanism of DCA on mastitis were evalued.The results showed that DCA dose dependently reduced the pathological damage of mouse breast tissue in-duced by Staphylococcus aureus;DCA inhibited the expression of inflammatory cytokines TNF-α and IL-1β and MPO activity induced by Staphylococcus aureus;DCA significantly reduced the con-centration of MDA and iron ions,while upregulating the expression of GSH and GPX4,indicating that DCA could inhibit ferroptosis induced by Staphylococcus aureus;Western blot results showed that DCA could upregulate TGR5 receptors,thereby inhibiting the NF-κB signaling pathway.The above results indicate that DCA inhibits mastitis by regulating the TGR5-NF-κB signaling pathway to inhibit the inflammatory response and ferroptosis induced by Staphylococcus aureus.

Objective:To investigate the correlation between cardiac polyps and gastroesophageal flap valve (GEFV).Methods:The clinical, endoscopic and pathological data of 349 patients with cardiac polyps (the cardiac polyp group) visiting Affiliated Hospital of Yangzhou University from January 1, 2016 to December 31, 2021 were retrospectively collected, and the same number of non-cardiac polyp patients (the non-cardiac polyp group) were matched in the same period as control according to the propensity score. The clinical, endoscopic and pathological data of the two groups were compared.Results:After matching with propensity score, there were 296 patients in each group, with no significant differences in smoking, acid reflux, heartburn, Helicobacter pylori infection, bile reflux, reflux esophagitis or pancreatitis between the two groups ( P>0.05). Compared with the non-cardiac polyp group, the risk of cardiac polyps increased in GEFV Ⅱ patients ( OR=3.046, 95%CI: 2.100-4.419, P<0.001) and GEFV Ⅲ patients ( OR=4.202, 95%CI: 2.299-7.681, P<0.001). Compared with the non-cardiac polyp group, the risk of cardiac polyps increased in patients with GEFV abnormalities ( OR=2.822, 95%CI: 1.615-4.931, P<0.001). GEFV abnormalities was associated with the cardiac polyp site ( χ2=22.169, P=0.003) and was not significantly associated with cardiac polyp size, number, morphology, intestinal metaplasia of the surrounding mucosa or intraepithelial neoplasia ( P>0.05). Conclusion:The occurrence of cardiac polyps is related to GEFV, and the patients with GEFV abnormalities are more likely to develop cardiac polyps.

ObjectiveTo investigate the mechanism of acupuncture in the treatment of chemotherapy-related cognitive impairment （CRCI） of breast cancer based on resting-state functional magnetic resonance imaging （rs-fMRI）. MethodTwenty-five patients with CRCI of breast cancer were included and treated with the acupuncture based on the method of regulating qi and blood， nourishing mind and benefiting intelligence； the selected acupoints included Zusanli （ST 36， bilateral）， Xuehai （SP 10， bilateral）， Tanzhong （CV 17）， Zhongwan （CV 12）， Qihai （CV 6）， Baihui （GV 20）， Fengfu （GV 16， bilateral）， Xinshu （BL 15， bilateral）， Tongli （HT 5， bilateral）， Zhaohai （KI 6， bilateral）， Yixi （BL 45， bilateral） twice a week， each time interval of 2-3 days， for 8 weeks. The scores of Montreal Cognitive Assessment （MoCA）， Mini-mental State Examination （MMSE）， European Cancer Research and Treatment Organization Quality of Life Questionnaire （EORTC QLQ-C30） were compared before and after acupuncture， and the effectiveness were evaluated by MoCA scale. The patients received rs-fMRI before and after treatment， and used low-frequency oscillation amplitude and functional connectivity （FC） analysis to extract the mean zALFF values of regions of interest such as bilateral anterior cingulate gyrus， bilateral hippocampus， bilateral amygdala， bilateral temporal pole middle temporal gyrus， and bilateral temporal pole supramarginal gyrus for comparison， and used the brain regions with statistically significant differences in the pre- and post-treatment zALFF values as the seed points for the seed-point-based FC Analysis. Correlation analyses were performed between the imaging metrics and the clinical scales. ResultsTwenty-four patients with CRCI of breast cancer completed treatment and follow-up. The zALFF values of the left hippocampus， left amygdala， and left temporal pole temporal gyrus in patients' rs-fMRI decreased after treatment （P＜0.05 or P＜0.01）， and left temporal pole temporal gyrus and right posterior cerebellar lobe FC were elevated （t = -5.169）， and MoCA scale total scores and visuospatial and executive， naming， and delayed recall cognitive scores， MMSE scale total scores， and EORTC QLQ-C30 scale mood scores were significantly higher （P＜0.05 or P＜0.01）. The total effective rate of MoCA was 58.33%. The difference in zALFF values of the left temporal pole middle temporal gyrus before and after treatment was negatively correlated with the MoCA total score （r= -0.499， P = 0.015）， as well as the difference in abstract function （r = -0.498， P = 0.016）. ConclusionThe acupuncture method of regulating qi and blood， nourishing mind and benefiting intelligence can improve the cognitive function of breast cancer patients with CRCI， and its mechanism may be related to improving the functional activities of hippocampus， amygdala and temporal lobe， as well as the functional connections of left temporal pole-temporal middle gyrus and right posterior cerebellar lobe.

Objective:To investigate the diagnostic performance of CT-derived fractional flow reserve (CT-FFR) derived from standard images (STD), images processed by first-generation (SSF1) and second-generation (SSF2) whole-heart motion correction algorithm, respectively.Methods:Patients who underwent both coronary CT angiography (CCTA) and invasive coronary angiography (ICA) with FFR examination within 3 months in Shanghai General Hospital, Shanghai Jiao Tong Univerisity School of Medicine from January 2020 to December 2022 were screened in this retrospective study. Totally of 121 patients (134 lesions) were finally included in the study. CCTA images were reconstructed using iterative reconstruction, iterative reconstruction plus SSF1 and SSF2 algorithms. All images were divided into three groups: STD group, SSF1 group, and SSF2 group. The image quality of the CCTA images was assessed using the Likert scale, and differences between the two groups were compared using the Mann-Whitney U and Kruskal-Wallis test. The correlation and consistency between CT-FFR and FFR were evaluated using Spearman correlation coefficient and Bland-Altman plots. The diagnostic performance of CCTA and CT-FFR from three groups was compared by receiver operating characteristic (ROC) curves. The area under the curve (AUC) was compared using the DeLong test. Results:Compared to the STD group and SSF1 group, the SSF2 group showed the best performance in image quality score (median=3.7). Best correlation ( r=0.652, P<0.001) and consistency (mean difference=0.03) between CT-FFR and FFR were observed in SSF2 group. ROC analysis results revealed that, at the per-lesion level, in the diagnosis of ischemic lesions, the diagnostic performance of CT-FFR in the SSF2 group was significantly better than that of the SSF1 group (AUC=0.88 vs. 0.76, P=0.003), while no significant difference was observed between STD group and SSF1 group ( P=0.125). At the per-patient level, the SSF2 group also demonstrated the highest diagnostic performance. Conclusion:The SSF2 algorithm significantly improved CCTA image quality and enhanced its diagnostic performance for evaluating stenosis severity and CT-FFR calculations.

Objective To investigate the effects of fibrin-derived peptide Bβ15-42(FgBβ15-42)on mitophagy in acute kidney injury(AKI)with ischemia-reperfusion at the cellular level.Methods Human kidney-2(HK2)cells were cultured in vitro and ran-domly divided into five groups:normal control group(Control group),hypoxia/reoxygenation(H/R)model group(H/R group),H/R+9μmol/L FgBβ15-42 peptide group(Fg group),H/R+20μmol/L chloroquine group(CQ group),and H/R+9μmol/L FgBβ15-42 peptide+20μmol/L chloroquine group(Fg+CQ group).Control group:cells were homogenized and replaced with complete medium and incubated in an aerobic incubator(5％CO2,21％O2)for 28hours.H/R group:cells were added to sugar-free and serum-free DMEM/F12medium and then placed in a hypoxic incubator(5％CO2,1％O2 and 94％N2)for 24hours,washed with PBS and added to DMEM/F12medium containing serum and then placed in a reoxygenated environment(5％CO2,21％O2)for 4hours.Fg group:mod-eled according to the above method,intervention was performed by adding FgBβ15-42 peptide 9μmol/L to the complete medium when reoxygenation was performed after the completion of hypoxia.CQ group:pre-treatment with chloroquine(20μmol/L)in serum-free medium at the time of homogenization,with the same post-procedural steps as in the H/R group.Fg+CQ group:the intervention was performed according to the dosing sequence of the CQ group and FgBβ15-42 group at different time periods as mentioned above,and the modeling time of the H/R group.After the intervention,CCK-8 was used to detect the cell viability of HK2.The mitochondrial morphol-ogy,mitophagosome structure and number were observed by electron microscope.The expression levels of autophagy related proteins p62 and LC3 were detected by Western blot.Results Compared with the control group,the survival rate of H/R group was decreased(P＜0.01);the mitochondrial damage was obvious,such as swelling and spinous rupture,and the number of mitophagosomes increased.The expression of autophagy protein LC-3 Ⅱ/Ⅰ was up-regulated(P＜0.05),and the expression of autophagy substrate protein p62 was down-regulated(P＜0.01).Compared with H/R group,the survival rate of Fg,CQ and Fg+CQ groups increased(P＜0.01),mito-phagosome was decreased.the expression of LC-3 Ⅱ/Ⅰ was down-regulated(P＜0.01),the expression of p62 was up-regulated(P＜0.05),and autophagy was inhibited.Conclusion The intervention of FgBβ15-42 peptide can reduce the level of autophagy and mitophagy in H/R-AKI cells,and has a protective effect on H/R injury in HK2 cells.

Objective:To explore the effect and mechanism of diosmetin (Dio) on neuronal ferroptosis in rats with bacterial meningitis (BM).Methods:Male SD rats aged 6-7 weeks of SPF grade were selected for the experiment. The BM model was established by injecting group B hemolytic streptococcus into the cisterna magna of cerebellum. Sixty BM model rats were successfully modeled and divided into model group, low-dose Dio group, medium-dose Dio group, high-dose Dio group and inhibitor group according to the random number table method, with 12 rats in each group. Another 12 weight-matched rats were taken as the control group.The rats in the low-dose Dio group, medium-dose Dio group, high-dose Dio group and the inhibitor group were intragastrically administered with Dio at 50 mg/kg, 100 mg/kg, 200 mg/kg and 200 mg/kg, respectively. The rats in the control group were intragastrically administered with an equal volume of 0.9 % sodium chloride solution. On the day of intragastric administration, the rats in the inhibitor group were intraperitoneally injected with SIRT1 pathway inhibitor EX527 (10 mg/kg), and the rats in the other groups were injected with an equal volume of 0.9% sodium chloride solution. The above interventions were performed once a day for 28 consecutive days. Loeffler neurological score was used to evaluate the neurological impairment in rats. Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cerebrospinal fluid of rats were detected by ELISA. The number of white blood cells in cerebrospinal fluid was detected by a blood cell analyzer. Glutathione (GSH) was detected by micro-enzyme labeling method, malondialdehyde (MDA) was detected by thiobarbituric acid colorimetric method, reactive oxygen species(ROS) was detected by colorimetry, and Fe 2+ level was detected by ferrozine method. Hematoxylin-eosin staining, Prussian blue staining and TUNEL staining were used to observe the pathological damage, iron accumulation and apoptosis in the hippocampus, respectively.Western blot was applied to measure the expression of transferrin (Tf), proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (Bax), caspase-3 and SIRT1/Nrf2/HO-1/Gpx4 signaling pathway proteins. Graphpad Prism 9.0 was used for data analysis. One-way ANOVA was used for statistical analysis, and SNK- q test was used for further pairwise comparisons. Results:(1) There was a statistically significant difference in neurological function scores among the 6 groups of rats ( F=125.451, P<0.001). The neurological function score of the model group was lower than that of control group, while the neurological function scores of the low-dose Dio group, medium-dose Dio group, and high-dose Dio group were higher than those of the model group (all P<0.05). The neurological function score of the inhibitor group ((2.57±0.26)) was lower than that of high-dose Dio group ((4.34±0.48)) ( P<0.05). (2) There were statistically significant differences in the levels of IL-6, TNF-α and the number of white blood cells in the cerebrospinal fluid of rats among the 6 groups ( F=127.817, 102.413, 180.967, all P<0.001). The levels of IL-6, TNF-α and the number of white blood cells in model group were higher than those of control group(all P<0.05). The levels of IL-6, TNF-α and the number of white blood cells in low-dose Dio group, medium-dose Dio group and high-dose Dio group were lower than those of model group (all P<0.001), and those in inhibitor group were all higher than those in high-dose Dio group(all P<0.001). (3) There were statistically significant differences in iron deposition rate and neuronal apoptosis rate among the 6 groups of rats ( F=90.857, 88.835, both P<0.001). The iron deposition rate ((18.37±3.14)%) and neuronal apoptosis rate ((27.58±2.63)%) in the inhibitor group were higher than those in the high-dose Dio group ((6.35±1.08)%, (14.02±1.87)%) (both P<0.05). (4) The levels of GSH, ROS, MDA, and Fe 2+ in the hippocampus of the 6 groups of rats showed statistically significant differences ( F=54.465, 106.453, 55.969, 105.457, all P<0.001). The GSH content in the inhibitor group ((103.48±8.76) mmol/g) was lower than that in the high-dose Dio group ((133.97±10.54) mmol/g), while the contents of ROS, MDA, Fe 2+ ((225.17±16.32) μmol/mg, (10.73±1.58) μmol/mg, (62.71±5.43) μg/g) were higher than those of the high-dose Dio group ((131.87±11.67) μmol/mg, (4.35±0.87) μmol/mg, (34.86±2.95) μg/g) (all P<0.05). (5)There were statistically significant differences in the protein levels of Tf, PCNA, Bax, caspase-3, SIRT1, Nrf2, HO-1 and Gpx4 in the hippocampus of the 6 groups of rats ( F=120.179, 107.568, 157.265, 98.031, 90.932, 52.283, 59.424, 114.539, all P<0.001). The protein levels of Tf, Bax and caspase-3 in the hippocampus of inhibitor group were higher than those of the high-dose Dio group, while the protein levels of PCNA, SIRT1, Nrf2, HO-1, Gpx4 were lower than those of the high-dose Dio group (all P<0.05). Conclusion:Diosmetin can activate SIRT1/Nrf2/HO-1/Gpx4 signaling pathway, thereby inhibiting neuronal ferroptosis in BM rats.

Objective: To investigate the incidence of microvascular myocardial ischemia in diabetic patients without obstructive coronary artery disease (CAD) and its relationship with angina. Materials and Methods: Diabetic patients and an intermediate-to-high pretest probability of CAD were prospectively enrolled. Non-diabetic patients but with an intermediate-to-high pretest probability of CAD were retrospectively included as controls. The patients underwent dynamic computed tomography-myocardial perfusion imaging (CT-MPI) and coronary computed tomography angiography (CCTA) to quantify coronary stenosis, myocardial blood flow (MBF), and extracellular volume (ECV). The proportion of patients with microvascular myocardial ischemia, defined as any myocardial segment with a mean MBF ≤ of 100 mL/min/100 mL, in patients without obstructive CAD (Coronary Artery Disease–Reporting and Data System [CAD-RADS] grade 0–2 on CCTA) was determined. Various quantitative parameters of the patients with and without diabetes without obstructive CAD were compared. Multivariable analysis was used to determine the association between microvascular myocardial ischemia and angina symptoms in diabetic patients without obstructive CAD. Results: One hundred and fifty-two diabetic patients (mean age: 59.7 ± 10.7; 77 males) and 266 non-diabetic patients (62.0 ± 12.3; 167 males) were enrolled; CCTA revealed 113 and 155 patients without obstructive CAD, respectively. For patients without obstructive CAD, the mean global MBF was significantly lower for those with diabetes than for those without (152.8 mL/min/100 mL vs. 170.4 mL/min/100 mL, P < 0.001). The mean ECV was significantly higher for diabetic patients (27.2% vs. 25.8%, P = 0.009). Among the patients without obstructive CAD, the incidence of microvascular myocardial ischemia (36.3% [41/113] vs. 10.3% [16/155], P < 0.001) and interstitial fibrosis (69.9% [79/113] vs. 33.3% [8/24], P = 0.001) were significantly higher in diabetic patients than in the controls. The presence of microvascular myocardial ischemia was independently associated with angina symptoms (adjusted odds ratio = 3.439, P = 0.037) in diabetic patients but without obstructive CAD. Conclusion Dynamic CT-MPI + CCTA revealed a high incidence of microvascular myocardial ischemia in diabetic patients without obstructive CAD. Microvascular myocardial ischemia is strongly associated with angina.