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.

The 2024 American Society of Clinical Oncology Gastrointestinal Cancers Symposium (ASCO-GI) was held in San Francisco, the USA from January 18th to 20th, 2024 (local time). The multiple studies presented in this symposium will have a significant impact on the clinical practice of esophageal cancer. This article will focus on the surgical methods of esophageal cancer, perioperative immunotherapy, drug therapy for advanced esophageal cancer, rescue treatment after immunotherapy resistance, and other relevant aspects. It aims to summarize and interpret the significant advancements in the field of esophageal cancer presented in this symposium.

Objective:To explore the efficacy and safety of single-incision transobturator bulbourethral sling suspension without skin tunnel puncture in male patients with urinary incontinence.Methods:The clinical data of 6 male patients with urinary incontinence who underwent single-incision transobturator bulbourethral sling suspension without skin tunnel puncture in Beijing Hospital from August 2023 to August 2024 were retrospectively analyzed.The age of the patients ranged from 66 to 76 years old, with an average of 71.7 years old. The disease duration ranged from 18 to 48 months, with an average of 30 months. Six patients used 1 to 3 pads per day, with an average of 2.3 pads. The International Continence Incontinence Questionnaire Short Form (ICI-Q-SF) scored 13 to 19, with an average of 15.8. The Incontinence Quality of Life Questionnaire (I-QOL) scored 5.3 to 30.6, with an average of 18.8. Three patients underwent transurethral resection of the prostate for benign prostatic hyperplasia and three patients underwent radical prostatectomy for prostate cancer. The degree of urinary incontinence was mild in 2 cases and moderate in 4 cases. The technical points are as follows: the puncture method has been changed from the traditional outside-in approach to an inside-out approach. After the puncture needle passes through from beneath the skin at the incision, the sling is guided in, avoiding the need for skin tunneling punctures. Upon completion of the puncture, the ends of the sling on both sides are tied with a certain tension at the midline of the incision, and the incision is then closed layer by layer. The efficacy and safety of surgery were evaluated by recording the number of daily pad use, subjective scoring scale [International Committee on Urinary Incontinence Questionnaire-Short Form (ICI-Q-SF), Incontinence Quality of Life (I-QOL)] and complications at 1 month after surgery. Social continence was defined as 0 to 1 pad use per day. Successful treatment was defined as social continence. Treatment improvement was defined as no social continence, but 50% or more improvement of symptoms compared with that before surgery. Other conditions were defined as treatment failure.Results:All operations were successfully completed. After 1 to 11 months of follow-up, all patients achieved social continence. The patients' postoperative daily use of urinary pads ranged from 0 to 1 piece, with a mean of 0.5 piece. ICI-Q-SF scores ranged from 1 to 7, with a mean of 3. I-QOL scores ranged from 72.1 to 85.2, with a mean of 77.0. All the indicators were significantly improved compared with those before operation. In terms of postoperative complications, one patient had dysuria and urinary retention 2 days after the removal of the catheter, which was improved after symptomatic treatment of anti-inflammatory, detumescence, and indwelling catheter. At the last follow-up, there were no surgical related complications.Conclusions:The single-incision transobturator bulbourethral sling suspension without skin tunnel puncture for the treatment of male urinary incontinence is safe and effective. Compared to the traditional surgical method, it does not increase the difficulty of the procedure and is technically feasible, offering clinicians a new approach and perspective.

Abstract: Approximately 2 billion people worldwide are infected with helminths and the resulting helminthiasis is a heavy health burden for developing countries. Parasitic helminths are a class of multicellular parasites, mainly including trematodes, tapeworms and nematodes, wtih complex life cycle involving multiple developmental stages and typically one or more hosts. Understanding the growth, development, pathogenesis and transmission of these parasites at the molecular level is of great significance for the diagnosis and treatment of helminthiasis. Genetic manipulations, which alter the expression level of target genes, have greatly promoted the biomedical research. In recent years, with the release of genomic data of worms, genetic manipulation techniques, such as RNA interference and clustered regularly interspaced short palindromic repeats (CRISPR) gene editing, have been increasingly applied in the studies of parasitic helminths. This article reviews the progress of genetic manipulations in important medical worms, as well as the methods of genetic manipulations, which would be expected to inspire the future functional study in parasitic helminths.

Background and Objectives: The search for a suitable alternative for urethral defect is a challenge in the field of urethral tissue engineering. Induced pluripotent stem cells (iPSCs) possess multipotential for differentiation. The in vitro derivation of urothelial cells from mouse-iPSCs (miPSCs) has thus far not been reported. The purpose of this study was to establish an efficient and robust differentiation protocol for the differentiation of miPSCs into urothelial cells. Methods: and Results: Our protocol made the visualization of differentiation processes of a 2-step approach possible. We firstly induced miPSCs into posterior definitive endoderm (DE) with glycogen synthase kinase-3β (GSK3β) inhibitor and Activin A. We investigated the optimal conditions for DE differentiation with GSK3β inhibitor treatment by varying the treatment time and concentration. Differentiation into urothelial cells, was directed with all-trans retinoic acid (ATRA) and recombinant mouse fibroblast growth factor-10 (FGF-10). Specific markers expressed at each stage of differentiation were validated by flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR) assay, immunofluorescence staining, and western blotting Assay. The miPSC-derived urothelial cells were successfully in expressed urothelial cell marker genes, proteins, and normal microscopic architecture. Conclusions We built a model of directed differentiation of miPSCs into urothelial cells, which may provide the evi-dence for a regenerative potential of miPSCs in preclinical animal studies.

【Objective】 To establish and verify the vacuum decay method for the tightness inspection of blood products. 【Methods】 The method for inspecting the tightness of blood product was established, and the detection limit, linearity, range, accuracy, precision and durability were verified according to the requirements of methodological verification.The validated method was used to check the tightness of blood product packaging. 【Results】 The detection limit of this method was 2.5 μm, linear correlation coefficient was r=1, the differential pressure of positive sample was within the allowable range of accuracy, and the durability met the requirements.The RSD of results of 6 repeatability tests and 12 intermediate precision tests were both less than 10%, and all validation items met the verification standards. 【Conclusion】 Vacuum decay method can be used to check the tightness of blood products.

Objective:To evaluate the effect of microsurgery for parafalcine meningioma through contralateral longitudinal fissure approach assisted by 3DSlicer software.Methods:From January 2020 to January 2021, 18 patients with parafalcine meningioma in The second hospital of Hebei Medical University were randomly selected as the observation group.The contralateral longitudinal fissure was treated by microsurgery assisted by 3DSlicer software.Before operation, improve the imaging examinations such as brain CT plain and enhancement, magnetic resonance angiography and magnetic resonance venography, establish the models of tumor, superior sagittal sinus and superior cerebral artery by using 3DSlicer software, analyze the positional relationship between tumor and superior sagittal sinus and superior cerebral vein, and remove the tumor by contralateral longitudinal fissure approach.Nineteen patients with parafalx meningioma treated by traditional ipsilateral longitudinal fissure approach from January to December 2019 were selected as the control group.The differences in operation time, intraoperative bleeding, limb muscle strength recovery rate, headache and dizziness relief rate were compared between the two groups.Results:In the observation group, the positional relationship between the reconstructed tumor and the superior cerebral vein above its base was completely consistent with the actual situation during the operation.All patients underwent Simpson grade I resection, and there was no injury to the superior cerebral vein and venous sinus during the operation.The recovery rate of limb muscle strength in the observation group (75%(9/12)) was higher than that in the control group (23%(3/13))( P=0.009). There was no significant difference in the operation time, intraoperative bleeding and the remission rate of headache and dizziness one week after operation in the observation group ( P>0.05). Conclusion:3DSlicer software was helpful for preoperative evaluation of the positional relationship between parafalcine meningioma and the anatomic structures such as superior cerebral vein and superior sagittal sinus.Under the guiding of 3DSlicer software, surgeon removed the tumor totally through the contralateral longitudinal fissure approach, which effectively reduced the operative side injury of the important structures such as superior cerebral vein and functional cerebral cortex, and contributed highly to the recovery of limb muscle strength of patients.

Objective To investigate the application prospect of the most extensive genome engineering pig internationally in preclinical xenotransplantation. Methods Porcine endogenous retrovirus (PERV) knockout combined with 3 major heterologous antigen gene knockouts and 9 humanized genes for inhibition of complement activation, regulation of coagulation disorders, anti-inflammatory and anti-phagocytosis were transferred into a pig (PERV-KO/3-KO/9-TG) as a donor, and the heart, liver and kidney were obtained and transplanted to 3 Rhesus macaque recipients respectively to establish a preclinical research model of pig-to-Rhesus macaque xenotransplantation. The functional status of xenografts after blood flow reconstruction was observed and the survival of recipients was summarized. The hemodynamics of xenografts were monitored. The change of hematological indexes of each recipient was compared. The histopathological manifestation of xenografts was observed. Results After the blood flow was reconstructed, all xenografts showed ruddy color, soft texture and good perfusion. The transplant heart, liver and kidney showed full arterial and venous blood flow and good perfusion at 1 d after operation. The postoperative survival time of heart, liver, and kidney transplant recipients was 7, 26, and 1 d, respectively. The levels of creatine kinase, creatine kinase isoenzyme, and lactate dehydrogenase increased in heart transplant recipient at 1 d after operation, and gradually recovered to near normal levels at 6 d after operation. All indexes increased sharply at 7 d after operation. The level of aspartate aminotransferase increased in liver transplant recipients at 2 d after operation, and the alanine aminotransferase basically returned to normal at 10 d after operation, but the total bilirubin continued to increase. Both aspartate aminotransferase and alanine aminotransferase increased at 12 d after operation, and reached a peak at 15 d after operation. The kidney transplant recipient developed mild proteinuria at 1 d after operation, and died of sudden severe arrhythmia. Histopathology showed that the tissue structure of cardiac and renal xenografts was close to normal, and liver xenografts presented with patchy necrosis, the liver tissue structure was disordered, accompanied by inflammatory damage, interstitial hemorrhage and thrombotic microangiopathy. Conclusions PERV-KO/3-KO/9-TG pig shows advantages in overcoming hyperacute rejection, mitigating humoral rejection and coagulation dysregulation. However, whether it can be used as potential donor for clinical xenotransplantation needs further evaluation.

Objective    To investigate the safety and feasibility of laryngeal mask general anesthesia as a replacement of tracheal intubation general anesthesia in the "three-port" thoracoscopic thymectomy via subxiphoid and subcostal arch for thymoma patients without myasthenia. Methods    From January 2018 to June 2019, clinical data of patients with thymoma who underwent the novel "three-port" operation in our institution were analyzed retrospectively. The patients were divided into two groups according to the anesthesia methods, including a tracheal intubation general anesthesia group and a laryngeal mask general anesthesia group. There were 70 patients in the tracheal intubation general anesthesia group, including 42 males and 28 females, with an average age of 45.83±15.89 years. There were 39 patients in the laryngeal mask general anesthesia group, including 26 males and 13 females, with an average age of 43.31±15.64 years. The clinical data of the two groups were compared. Results    The baseline characteristics of the patients in the two groups were well balanced (P>0.05). No massive bleeding, conversion to thoracotomy, postoperative myasthenia or death occurred in those patients. No patient with laryngeal mask anesthesia had a conversion to tracheal intubation anesthesia during the operation. There was no significant difference in the operation time, intraoperative bleeding, intraoperative maximum partial pressure of CO2, lowest partial pressure of oxygen and anesthesia effect score between the two groups (P>0.05). There was also no statistical difference in postoperative aspiration, gastrointestinal discomfort, length of hospital stay, pain score and patient satisfaction degree between the two groups (P>0.05). However, the anesthesia time before operation and the time of awake after anesthesia in the laryngeal mask anesthesia group were significantly shorter than those in the tracheal intubation general anesthesia group (P<0.05), and the incidence of transient arrhythmia, laryngeal discomfort and hoarseness in the laryngeal mask general anesthesia group was significantly lower than that in the tracheal intubation general anesthesia group (P<0.05). Conclusion    The "three-port" thoracoscopic thymectomy via subxiphoid and subcostal arch under laryngeal mask general anesthesia is safe and feasible in the treatment of thymoma without myasthenia, and can be recommended routinely.

Objective    To evaluate the clinical outcomes of larynx-preserving limited resection with total thoracic esophagectomy and gastric pull-up reconstruction for the treatment of cervical esophageal squamous cell carcinoma (ESCC) without tumor involvement of the larynx and hypopharynx compared with the upper thoracic ESCC. Methods    Retrospective and comparative analysis of consecutive patients with cervical and upper thoracic ESCC who underwent R0 surgical resection from 2006 to 2011 in our center was performed. Kaplan-Meier method was used to calculate the patients’ survival. Results    In total, 44 pairs of patients, including 71 males and 17 females with an average age of 60.66±8.49 years were enrolled in the study after propensity score matching. The baseline characteristics of the two groups of patients were well balanced. There was no statistical difference in the operation time (P=0.100), blood loss (P=0.685), mortality rate in 30 days (P=1.000), total complication rate (P=0.829), cervical anastomosis leakage (P=0.816), mechanical ventilation (P=1.000), normal oral diet within 15 days (P=0.822) and anastomosis recurrence rate (P=0.676) between the two groups. Survival analysis showed that there was no statistical difference in survival time between the cervical group [31.83 (95%CI 8.65-55.02) months] and upper thoracic group [37.73 (95%CI 25.29-50.18) months, P=0.533]. The 5-year survival rates were 32.6% and 42.1%, respectively. Conclusion    Larynx-preserving limited resection  with total thoracic esophagectomy and gastric pull-up reconstruction for the treatment of cervical ESCC without involvement of the larynx and hypopharynx may result in a similar clinical outcome to upper thoracic ESCC.