OBJECTIVE To explore the mechanism of gallic acid targeting β-arrestin2 to inhibit astrocyte inflammation.METHODS Potential targets of gallic acid were found by PharmMapper and verified by thermal shift and molecular docking experi-ments.Western blot was used to detect the expression of β-arrestin2,NF-κB signaling pathway and NLRP3 inflammasome-related proteins;qPCR was used to detect the mRNA levels of proinflammatory cytokines such as IL-1β,IL-6 and TNF-α.A subacute PD mouse model induced by MPTP was established,and the behavioral ability of mice was evaluated by open field test,rotating rod test and climbing pole test;the number of TH+and GFAP+cells in the SNc area of mice was detected by immunohistochemistry;Western blot was used to detect the expression of NF-κB signaling pathway and NLRP3 inflammasome-related proteins in the homogenate pro-tein of mouse midbrain tissue.RESULTS Molecular docking experiments and thermal shift experiments showed that gallic acid could directly bind to β-arrestin2.Gallic acid could reduce the expression of p-IKK and p-P65 proteins in astrocytes(P<0.001,P<0.000 1),reduce the mRNA levels of IL-1β,IL-6 and TNF-α(P<0.05,P<0.01),and reduce the expression of caspase-1 and IL-1β proteins(P<0.000 1),but had no effect on the expression of β-arrestin2 protein(P>0.05).Gallic acid could improve the be-havioral ability of PD model mice,increase the number of TH+neurons and GFAP+cells in PD model mice(P<0.05,P<0.001),and reduce the expression of caspase-1,IL-1β,NLRP3 and p-IKK proteins in the brain of PD model mice(P<0.05).CONCLU-SION Gallic acid inhibits the activation of NF-κB signaling pathway and NLRP3 inflammasome by binding to β-arrestin2 to reduce astrocyte inflammation,and has a neuroprotective effect on MPTP-induced PD model mice.

Objective To investigate the anti-hepatocellular carcinoma mechanism of Chloranthus fortunei(A.Gray)Solms-Laub.via network pharmacology,molecular docking techniques and in vitro experiments.Methods Chemical composition of Chloranthus fortunei(A.Gray)Solms-Laub.was searched by literature.Swiss Target Prediction was used to find corresponding targets.STRING was used to construct protein-protein interactions network(PPI).DAVID was used to enrich GO analysis and KEGG pathway.AutoDock Vina 1.1.2 and Pymol visualisation was used for docking and validation.Results Chloranthus fortunei(A.Gray)Solms-Laub.had 61 active components,685 targets,and 279 intersections with disease targets.The PPI showed that the main active components were Luteolin,Chloranthalactone C,Shizukanolide H,Esculetin,7-Hydroxycoumarin.The key targets were GAPDH,VEGFA,STAT3,JUN,HSP90AA1,AKT1,CTNNB1,CASP3,and ALB.Biological process(BP)involved protein phosphorylation,signal transduction,regulation of RNA polymerase II promoter transcription,cell proliferation,apoptosis.Cellular component(CC)involved cytoplasm,nucleus,cell membrane,cellular exosome.Molecular function(MF)involves protein binding,ATPase,threonine kinase,protein kinase activity.KEGG involved cancer pathway,metabolic pathway,PI3K-Akt signalling pathway,cancer proteoglycans,lipids and atherosclerosis,cytomegalovirus infection,microRNAs in cancer,human T-cell leukaemia virus type 1,Ras signalling pathway,MAPK signalling pathway.Molecular docking showed that silverweed lactone H had a strong affinity for each of the other target proteins,indicating that this component plays a key role.The results of RT-qPCR assay and WB assay showed that there were significant differences in gene and protein expression levels before and after drug administration.Conclusion The Chinese medicine in Chloranthus fortunei(A.Gray)Solms-Laub.can treat hepatocellular carcinoma through the MAPK pathway,and the main active ingredients have good docking effects with the core target proteins of the disease.

Objective To explore the short-term efficacy of endoscopic submucosal dissection (ESD) in the treatment of early carcinoma in the remnant stomach. Methods A retrospective study was conducted on 45 patients with early residual gastric cancer underwent ESD at the Endoscopy Center of Zhongshan Hospital, Fudan University from December 2014 to April 2024, with a total of 45 lesions. The patients were divided into an anastomotic group (n=15) and a non-anastomotic group (n=30) based on the location of tumor occurrence, and their clinical data, endoscopic diagnosis and treatment, and histopathological conditions were compared between the two groups. Results All 45 patients had lesions with redness and erosion. There were 9 cases of poor lifting of submucosal injection in the anastomotic group and 2 cases in the non-anastomotic group, respectively, and the difference was statistically significant (P<0.05). ESD surgery was performed on 13 lesions in the anastomotic group and 28 lesions in the non-anastomotic group, with surgery times of 80.00 (50.00, 100.00) min and 55.00 (43.75, 80.00) min, respectively. The difference in surgery time between the two groups was statistically significant (P=0.03). Among the 45 patients, ESD surgery achieved curative resection in 35 cases, including 11 cases in the anastomotic group and 24 cases in the non-anastomotic group, with no statistically significant difference. Conclusions Careful preoperative evaluation of early carcinoma in the remnant stomach is essential to prevent oversight. Lesions at anastomotic sites and suture lines present higher technical challenges for complete resection. ESD is safe and effective, with auxiliary traction technique available when necessary.

OBJECTIVE To explore the mechanism of gallic acid targeting β-arrestin2 to inhibit astrocyte inflammation.METHODS Potential targets of gallic acid were found by PharmMapper and verified by thermal shift and molecular docking experi-ments.Western blot was used to detect the expression of β-arrestin2,NF-κB signaling pathway and NLRP3 inflammasome-related proteins;qPCR was used to detect the mRNA levels of proinflammatory cytokines such as IL-1β,IL-6 and TNF-α.A subacute PD mouse model induced by MPTP was established,and the behavioral ability of mice was evaluated by open field test,rotating rod test and climbing pole test;the number of TH+and GFAP+cells in the SNc area of mice was detected by immunohistochemistry;Western blot was used to detect the expression of NF-κB signaling pathway and NLRP3 inflammasome-related proteins in the homogenate pro-tein of mouse midbrain tissue.RESULTS Molecular docking experiments and thermal shift experiments showed that gallic acid could directly bind to β-arrestin2.Gallic acid could reduce the expression of p-IKK and p-P65 proteins in astrocytes(P<0.001,P<0.000 1),reduce the mRNA levels of IL-1β,IL-6 and TNF-α(P<0.05,P<0.01),and reduce the expression of caspase-1 and IL-1β proteins(P<0.000 1),but had no effect on the expression of β-arrestin2 protein(P>0.05).Gallic acid could improve the be-havioral ability of PD model mice,increase the number of TH+neurons and GFAP+cells in PD model mice(P<0.05,P<0.001),and reduce the expression of caspase-1,IL-1β,NLRP3 and p-IKK proteins in the brain of PD model mice(P<0.05).CONCLU-SION Gallic acid inhibits the activation of NF-κB signaling pathway and NLRP3 inflammasome by binding to β-arrestin2 to reduce astrocyte inflammation,and has a neuroprotective effect on MPTP-induced PD model mice.

Objective To investigate the anti-hepatocellular carcinoma mechanism of Chloranthus fortunei(A.Gray)Solms-Laub.via network pharmacology,molecular docking techniques and in vitro experiments.Methods Chemical composition of Chloranthus fortunei(A.Gray)Solms-Laub.was searched by literature.Swiss Target Prediction was used to find corresponding targets.STRING was used to construct protein-protein interactions network(PPI).DAVID was used to enrich GO analysis and KEGG pathway.AutoDock Vina 1.1.2 and Pymol visualisation was used for docking and validation.Results Chloranthus fortunei(A.Gray)Solms-Laub.had 61 active components,685 targets,and 279 intersections with disease targets.The PPI showed that the main active components were Luteolin,Chloranthalactone C,Shizukanolide H,Esculetin,7-Hydroxycoumarin.The key targets were GAPDH,VEGFA,STAT3,JUN,HSP90AA1,AKT1,CTNNB1,CASP3,and ALB.Biological process(BP)involved protein phosphorylation,signal transduction,regulation of RNA polymerase II promoter transcription,cell proliferation,apoptosis.Cellular component(CC)involved cytoplasm,nucleus,cell membrane,cellular exosome.Molecular function(MF)involves protein binding,ATPase,threonine kinase,protein kinase activity.KEGG involved cancer pathway,metabolic pathway,PI3K-Akt signalling pathway,cancer proteoglycans,lipids and atherosclerosis,cytomegalovirus infection,microRNAs in cancer,human T-cell leukaemia virus type 1,Ras signalling pathway,MAPK signalling pathway.Molecular docking showed that silverweed lactone H had a strong affinity for each of the other target proteins,indicating that this component plays a key role.The results of RT-qPCR assay and WB assay showed that there were significant differences in gene and protein expression levels before and after drug administration.Conclusion The Chinese medicine in Chloranthus fortunei(A.Gray)Solms-Laub.can treat hepatocellular carcinoma through the MAPK pathway,and the main active ingredients have good docking effects with the core target proteins of the disease.

Objective:To explore a simplified and efficient early prediction model for severe acute pancreatitis (SAP) using the least absolute shrinkage and selection operator (LASSO) regression, and to construct both logistic regression and decision tree models. The aim is to identify high-risk individuals, guide clinical treatment, and improve patient outcomes.Methods:A retrospective analysis was conducted on the clinical data of 412 patients with acute pancreatitis admitted to the Emergency and Gastroenterology Departments of the First Affiliated Hospital of Anhui Medical University and its High-tech Branch from November 2020 to September 2023. LASSO regression was employed to identify factors significantly associated with SAP, followed by the construction of a multivariate logistic regression model and a decision tree model. The predictive performance of these models was evaluated and compared to the bedside index for severity in acute pancreatitis (BISAP).Results:Among the 412 patients, the incidence of SAP was 12.14% ( n=50). Seven variables significantly associated with SAP severity were identified by LASSO regression, including respiratory rate at admission, pain score at admission, pleural effusion, fibrin degradation products, C-reactive protein, serum creatinine, and serum albumin. The logistic regression model incorporated four variables: pleural effusion, pain score at admission, serum creatinine, and serum albumin. In the training set, the model demonstrated a sensitivity of 0.528, specificity of 0.984, accuracy (95% CI) of 0.928 (0.892-0.955), Kappa value of 0.606, and AUC (95% CI) of 0.920 (0.862-0.979). In the testing set, the model showed a sensitivity of 0.643, specificity of 0.925, accuracy (95% CI) of 0.891 (0.822-0.941), Kappa value of 0.519, and AUC (95% CI) of 0.923 (0.861-0.985). The decision tree model comprised three branches and four terminal nodes, indicating that serum creatinine, serum albumin, and pleural effusion could effectively predict SAP occurrence. In the training set, the decision tree model had a sensitivity of 0.500, specificity of 0.973, accuracy (95% CI) of 0.914 (0.876-0.944), Kappa value of 0.544, and AUC (95% CI) of 0.812 (0.731-0.894). In the testing set, the model exhibited a sensitivity of 0.500, specificity of 0.925, accuracy (95% CI) of 0.875 (0.802-0.928), Kappa value of 0.412, and AUC (95% CI) of 0.709 (0.565-0.853). The DeLong test revealed that in the training set, the AUC of the logistic regression model was significantly greater than that of the decision tree model ( P<0.01) and the BISAP score ( P<0.001), while the AUC difference between the decision tree model and the BISAP score was not statistically significant ( P=0.762). In the testing set, the AUC of the logistic regression model was again greater than that of the decision tree model ( P<0.01) and the BISAP score ( P=0.018), whereas the AUC of the decision tree model was lower than that of the BISAP score ( P=0.017). Conclusions:Both the logistic regression and decision tree models demonstrate good predictive value for SAP, and their combined use may provide valuable guidance for clinical practice.

Objective:To carry out an overview of systematic reviews on interventions to prevent the recurrence of diabetic foot ulcer (DFU) patients.Methods:Systematic reviews or Meta-analysis on interventions to prevent DFU recurrence were electronically retrieved from databases such as China National Knowledge Infrastructure, WanFang Data, VIP, China Biology Medicine disc, Joanna Briggs Institute (JBI) Evidence-Based Healthcare Center Database, Cochrane Library, PubMed, Embase, CINAHL and Web of Science. The search period was from database establishment to April 20, 2023. Two researchers independently searched and screened literature, and extracted data, and used AMSTAR 2 software and the quality evaluation criteria for systematic review of JBI Evidence-Based Healthcare Center to evaluate the methodological quality of the included literature. GRADE evaluation system was used for quality assessment of outcome indicators (DFU recurrence rate, effectiveness of measures to prevent DFU recurrence) .Results:A total of 24 systematic reviews were included. Studies showed that monitoring and intervention of foot skin temperature, therapeutic shoes or insoles, comprehensive intervention measures, and specific surgical methods could reduce the recurrence rate of DFU, while foot self-care, foot exercise, health education, and psychological intervention had no statistical effect on preventing DFU recurrence. The methodological quality of systematic reviews was generally low. The reports of four articles were relatively complete, 18 articles had certain defects, and two articles had serious defects. The evaluation of evidence quality showed that there were three pieces of moderate-quality evidence, seven pieces of low-quality evidence, and 30 pieces of extremely low-quality evidence.Conclusions:Existing evidence suggests that foot skin temperature monitoring and intervention, therapeutic shoes or insoles, comprehensive intervention measures, and specific surgical methods (such as Achilles tendon lengthening, metatarsophalangeal joint replacement, metatarsal head resection and so on) have a positive impact on preventing DFU recurrence. Rigorous and high standard research is still needed to verify the controversial issues, so as to provide reliable evidence for future clinical practice and studies.

Objective:To investigate the perioperative risk factors in contribution of hyperbilirubinemia following cardiopulmonary bypass(CPB) assisted cardiovascular surgery, of which cutoff values of key factors are defined.Methods:1 286 patients received cardiac surgery assisted by CPB from January 2017 to March 2019 were included in the study. The perioperative data and the peak serum total bilirubin at selected timepoints were recorded. Logistic regression of multi-factor analysis was used to define risk factors and then broken-line analysis was applied to predict the risky threshold. Results:312(24.26%)patients developed hyperbilirubinemia after surgery, with the in-hospital mortality rate up to 34.62%(108 cases). In those patients, valve surgery(45.51%, 142/312), great vessel open surgery(37.82%, 118/312) and heart transplantation(7.69%, 24/312) were mostly performed. The duration of postoperative ICU stay and the use of ventilation were 6 days and 68 hours, which were significantly higher than those in non-hyperbilirubinemia group( P<0.01). Multivariate logistic regression showed that the postoperative ventilation time>49 h, the cardiopulmonary bypass(CPB) time>181 min and the abnormal preoperative liver function, use of intra-aortic balloon pump and extracorporeal membrane oxygen, unplanned re-exploration for bleeding were the risk factors for postoperative hyperbilirubinemia( P<0.01). The cutoff duration of postoperative mechanical ventilation and CPB affecting the incidence of hyperbilirubinemia, ICU days and in-hospital mortality were 120.21 h and 143.26 min, 248.20 h and 239.51 min, 259.50 h and 190.60 min, respectively. Conclusion:Preoperative abnormal liver function, intraoperative CPB time, postoperative mechanical ventilation time, postoperative use of IABP or ECMO assistance, and unplanned secondary thoracotomy were high-risk factors for hyperbilirubinemia after CPB-assisted cardiovascular surgery.

Cullin-RING E3 ubiquitin ligase (CRL)-4 is a member of the large CRL family in eukaryotes. It plays important roles in a wide range of cellular processes, organismal development, and physiological and pathological conditions. DDB1- and CUL4-associated factor 8 (DCAF8) is a WD40 repeat-containing protein, which serves as a substrate receptor for CRL4. The physiological role of DCAF8 is unknown. In this study, we constructed Dcaf8 knockout mice. Homozygous mice were viable with no noticeable abnormalities. However, the fertility of Dcaf8-deficient male mice was markedly impaired, consistent with the high expression of DCAF8 in adult mouse testis. Sperm movement characteristics, including progressive motility, path velocity, progressive velocity, and track speed, were significantly lower in Dcaf8 knockout mice than in wild-type (WT) mice. However, the total motility was similar between WT and Dcaf8 knockout sperm. More than 40% of spermatids in Dcaf8 knockout mice showed pronounced morphological abnormalities with typical bent head malformation. The acrosome and nucleus of Dcaf8 knockout sperm looked similar to those of WT sperm. In vitro tests showed that the fertilization rate of Dcaf8 knockout mice was significantly reduced. The results demonstrated that DCAF8 plays a critical role in spermatogenesis, and DCAF8 is a key component of CRL4 function in the reproductive system.
Animals ; Cullin Proteins/genetics* ; DNA-Binding Proteins/genetics* ; Factor VIII ; Male ; Mice ; Mice, Knockout ; Spermatogenesis/genetics* ; Ubiquitin-Protein Ligases

Objective:To compare the clinical effects of 4 kinds of neurocutaneous perforator flap with vascular anastomosis for repair of hand and foot wounds.Methods:From January, 2005 to September, 2019, 112 patients with hand and foot wounds were treated, there were 78 cases of fingers, 11 cases of first web, 5 cases of palm, 6 cases of hand and 12 cases of foot. The defect area was 2.0 cm×1.5 cm-21.0 cm×12.0 cm. All 112 cases were repaired by neurocutaneous perforator flaps anastomosed with blood vessels. Types of flap were applied: Radial collateral artery perforator flap (with posterior cutaneous nerve of forearm) in 30 cases. The flap area was 5.0 cm×2.0 cm-13.0 cm×6.0 cm. Superficial peroneal artery flap (without superficial peroneal nerve) anastomosed with blood vessels in 15 cases. The flap area was 2.5 cm×2.0 cm-9.0 cm×6.0 cm. Lateral superficial sural artery perforator flap (with superior sural cutaneous nerve) in 26 cases. The flap area was 2.5 cm×1.8 cm-7.0 cm×5.0 cm. Peroneal artery perforator flap (with middle and lower sural nerve) in 41 cases. The flap was harvested with area of 2.5 cm×1.8 cm-23.0 cm ×14.0 cm to repair the wounds of feet, back of hands, first web, palm and fingers. CTA images were observed in 40 clinical patients, and the occurrence rate of radial collateral artery, superficial peroneal artery, superficial lateral sural artery, and peroneal artery were measured. Anastomosis cutaneous nerve in 97 cases, and no cutaneous nerve anastomosis 15 cases (superficial peroneal artery flap).Results:The peroneal artery perforator flap (41 cases) and radial collateral artery perforator flap (30 cases) were harvested. The incidence of perforator vessels was both 100%, and incidence of superficial sural artery was 80.8% (21/26 cases). In the other 19.2% (5/26 cases), the superficial medial sural artery was replaced by too thin vessels. The utilization rate of superficial peroneal artery was 60.0% (9/15 cases), the other 40.0% (6/15 cases) were converted to peroneal artery perforator flap. All flaps survived except 1 case of superficial perforator flap of lateral sural artery, which underwent necrosis at the distal end and healed after dressing change. One hundred and one cases were followed-up, including 90 cases for repairing soft tissue defects in hands and 11 cases in feet. The followed-up time ranged from 12 to 120 months, with an average of 36.6 months. There were 40 cases with excellent function, 45 cases with good function and 5 cases with fair function. There were 78 cases of cutaneous nerve anastomosis of hand flap, and the sensory function was above S 3 level. There were 12 cases without anastomosis of cutaneous nerve of hand flap, and the sensory function reached S 3 level in 3 cases and S 2 level in 9 cases. In 11 cases, the cutaneous nerve was anastomosed to repair the soft tissue defect of the foot, and the sensory function was above S 3 level. The radial collateral artery perforator flaps were relatively bulky and needed to be treated by fat removal. The other 3 kinds of three flaps were not bulky. Conclusion:The perforating vessels of peroneal artery and radial accessory artery have larger diameter and easy to harvest. The superficial peroneal artery and the lateral superficial sural artery are relatively small in caliber, especially the superficial peroneal artery. Among the 4 kinds of cutaneous nerve nutrient vascular flaps, the radial accessory artery perforator flap was the most bloated. Sensory nerve innervation flaps were found in the upper segment of lateral sural cutaneous nerve, posterior forearm cutaneous nerve and middle and lower segment of sural nerve. The superficial peroneal artery perforator flap was accompanied by superficial peroneal nerve that did not send cutaneous branches into the flap. The upper segment of superficial peroneal nerve was only a passing nerve.