Background and Aims:Reconstructing huge abdominal wall defects after abdominal wall tumor resection is a significant challenge for surgeons.Here,we summarize the experience of our team in repairing these defects using an anterolateral thigh chimeric perforator flap combined with artificial materials,within a multidisciplinary collaborative framework,aiming to provide a reference for other practitioners.Methods:The clinicopathologic data of 7 patients who underwent primary reconstruction of large abdominal wall defects after malignant tumor resection,using an anterolateral thigh chimeric perforator flap combined with artificial materials,at Xiangya Hospital,Central South University,between October 2019 and April 2024,were retrospectively analyzed.The clinical outcomes were evaluated.Results:All 7 patients successfully underwent both tumor resection and primary abdominal wall reconstruction without surgery-related mortality.The wound in all patients healed in one stage,with no severe complications such as surgical site infections or flap necrosis.During a follow-up period of 6 to 54 months,2 patients experienced tumor recurrence,one of whom died(exact cause unknown),while the other continued anti-tumor treatment.The remaining 5 patients were alive without recurrence.One patient developed abdominal wall bulging,but no obvious incisional hernia occurred in any of the patients.Conclusion:The use of an anterolateral thigh chimeric perforator flap combined with artificial materials for repairing large abdominal wall defects after abdominal tumor resection can achieve satisfactory therapeutic outcomes.This approach is a feasible and safe method for abdominal wall reconstruction.

Background and Aims:Early-onset extensive emphysematous pancreatitis(EP)is a rare but highly lethal subtype of infected pancreatic necrosis(IPN),characterized by abrupt onset and rapid deterioration.This study aimed to investigate its clinical characteristics,microbiological spectrum,treatment approaches,and outcomes to provide evidence for early identification and timely intervention.Methods:A retrospective analysis was performed on 305 IPN patients treated at Xiangya Hospital,Central South University,from January 2010 to October 2023.Eight patients who developed gas accumulation involving≥50%of pancreatic or peripancreatic necrosis within two weeks of onset were defined as early-onset extensive EP.Their clinical data were compared with those of ordinary IPN patients.Results:Early-onset extensive EP accounted for 2.6%of all IPN cases.The early-onset extensive EP group had significantly higher mortality and multiple organ failure rates compared with the ordinary IPN group(75.0%vs.24.6%and 75.0%vs.34.7%,respectively;both P<0.05).A total of 15 microbial isolates were identified from early-onset extensive EP patients,predominantly Klebsiella pneumoniae(62.5%)and Escherichia coli(37.5%).The infection rate of carbapenem-resistant Enterobacteriaceae(CRE)was markedly higher in the EP group than in the ordinary IPN group(75.0%vs.31.1%,P=0.015).Most patients were treated using a step-up approach based on percutaneous catheter drainage,with no significant difference in treatment strategy between the two groups(P=0.625).Conclusion:Early-onset extensive EP represents a rare and fulminant subtype of IPN with extremely poor outcomes.Klebsiella pneumoniae and CRE are the predominant pathogens.Early radiological evaluation and timely intervention are crucial for improving prognosis in these patients.

Background and Aims:Reconstructing huge abdominal wall defects after abdominal wall tumor resection is a significant challenge for surgeons.Here,we summarize the experience of our team in repairing these defects using an anterolateral thigh chimeric perforator flap combined with artificial materials,within a multidisciplinary collaborative framework,aiming to provide a reference for other practitioners.Methods:The clinicopathologic data of 7 patients who underwent primary reconstruction of large abdominal wall defects after malignant tumor resection,using an anterolateral thigh chimeric perforator flap combined with artificial materials,at Xiangya Hospital,Central South University,between October 2019 and April 2024,were retrospectively analyzed.The clinical outcomes were evaluated.Results:All 7 patients successfully underwent both tumor resection and primary abdominal wall reconstruction without surgery-related mortality.The wound in all patients healed in one stage,with no severe complications such as surgical site infections or flap necrosis.During a follow-up period of 6 to 54 months,2 patients experienced tumor recurrence,one of whom died(exact cause unknown),while the other continued anti-tumor treatment.The remaining 5 patients were alive without recurrence.One patient developed abdominal wall bulging,but no obvious incisional hernia occurred in any of the patients.Conclusion:The use of an anterolateral thigh chimeric perforator flap combined with artificial materials for repairing large abdominal wall defects after abdominal tumor resection can achieve satisfactory therapeutic outcomes.This approach is a feasible and safe method for abdominal wall reconstruction.

Background and Aims:Early-onset extensive emphysematous pancreatitis(EP)is a rare but highly lethal subtype of infected pancreatic necrosis(IPN),characterized by abrupt onset and rapid deterioration.This study aimed to investigate its clinical characteristics,microbiological spectrum,treatment approaches,and outcomes to provide evidence for early identification and timely intervention.Methods:A retrospective analysis was performed on 305 IPN patients treated at Xiangya Hospital,Central South University,from January 2010 to October 2023.Eight patients who developed gas accumulation involving≥50%of pancreatic or peripancreatic necrosis within two weeks of onset were defined as early-onset extensive EP.Their clinical data were compared with those of ordinary IPN patients.Results:Early-onset extensive EP accounted for 2.6%of all IPN cases.The early-onset extensive EP group had significantly higher mortality and multiple organ failure rates compared with the ordinary IPN group(75.0%vs.24.6%and 75.0%vs.34.7%,respectively;both P<0.05).A total of 15 microbial isolates were identified from early-onset extensive EP patients,predominantly Klebsiella pneumoniae(62.5%)and Escherichia coli(37.5%).The infection rate of carbapenem-resistant Enterobacteriaceae(CRE)was markedly higher in the EP group than in the ordinary IPN group(75.0%vs.31.1%,P=0.015).Most patients were treated using a step-up approach based on percutaneous catheter drainage,with no significant difference in treatment strategy between the two groups(P=0.625).Conclusion:Early-onset extensive EP represents a rare and fulminant subtype of IPN with extremely poor outcomes.Klebsiella pneumoniae and CRE are the predominant pathogens.Early radiological evaluation and timely intervention are crucial for improving prognosis in these patients.

Objective:To investigate the application value of metagenomic next-genera-tion sequencing (mNGS) in pathogenic diagnosis of suspected infected severe acute pancreatitis (SAP).Methods:The prospective study was conducted. The clinical data of 25 patients with suspected infected SAP who were admitted to the Xiangya Hospital of Central South University from May to September 2023 were collected. Upper limb venous blood samples of all the patients were collected for both of mNGS and routine pathogen microbial culture. Observation indicators: (1) grouping situations of the enrolled patients; (2) comparison of the diagnostic efficiency of mNGS and routine pathogen microbial culture; (3) results of peripheral blood pathogen microbial testing and peri-pancreatic effusion microbial culture; (4) testing time and cost. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3). Count data were expressed as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Results:(1) Grouping situations of the enrolled patients. A total of 25 patients were selected for eligibility. There were 18 males and 7 females, aged 48(40,59)years. The duration of hospital stay of 25 patients was 30(20,50)days. The etiologies of 25 patients included 14 cases of hyperlipidemic pancreatitis, 8 cases of biliary pancreatitis, 1 case of alcohol-induced acute pancreatitis, and 2 cases of pancreatitis caused by other causes. Of the 25 patients, there were 17 cases with infected pancreatic necrosis (IPN) including 7 cases of death, and 8 cases with sterile pancreatic necrosis including no death. (2) Comparison of the diagnostic efficiency of mNGS and routine pathogen microbial culture. The positive rates of mNGS and routine pathogen microbial culture in diagnosis of suspected infected SAP were 72.0%(18/25) and 32.0%(8/25), respectively, showing a significant difference between them ( χ2=8.01, P<0.05). The sensitivity and negative predic-tive value of mNGS and routine pathogen microbial culture in diagnosis of IPN were 94.1%(16/17), 35.3%(6/17) and 85.7%(6/7), 35.3%(6/17), showing significant differences between them ( χ2=12.88, 5.04, P<0.05). The specificity and positive predictive value of mNGS and routine pathogen microbial culture in diagnosis of IPN were 75.0%(6/8), 75.0%(6/8) and 88.9%(16/18), 75.0%(6/8), showing no significant difference between them ( χ2=0, 0.82, P>0.05). (3) Results of peripheral blood pathogen microbial testing and peripancreatic effusion microbial culture. Of the 17 patients with IPN, 36 strains of pathogenic bacteria were detected by mNGS, and 6 strains were detected by routine pathogen microbial culture. There were 16 of 17 patients with IPN showing positive mNGS pathogenic testing, of which 13 cases were consistent with the pathogenic testing results of peri-pancreatic effusion microbial culture, showing a consistency rate of 76.5%(13/17). There were 6 pati-ents with IPN showing positive routine pathogen microbial culture, with a consistency rate of 35.3%(6/17) to peripancreatic effusion microbial culture. (4) Testing time and cost. Testing time of mNGS and routine pathogen microbial culture were (43±17)hours and (111±36)hours, showing a signifi-cant difference between them ( t=9.31, P<0.05). Testing cost of mNGS was (2 267±0)yuan/case, accoun-ting for 1.7% of the hospitalization expenses of (133 759±120 744)yuan/case. Testing cost of routine pathogen microbial culture was (240±0)yuan/case, accounting of 0.2% of the hospitalization expenses. Conclusion:mNGS has important value for early pathogenic diagnosis of suspected infected SAP, and has a high timeliness.

Background and Aims:Accurate early pathogen diagnosis is a breakthrough for improving the prognosis of infectious pancreatic necrosis(IPN)patients.However,there is currently a lack of efficient methods for early identification of IPN in clinical settings.This study was performed to assess the application value of next-generation sequencing technology based on metagenomic capture(MetaCAP)in the pathogen diagnosis of IPN. Methods:A prospective study was conducted on 29 patients suspected of having acute necrotizing pancreatitis at Xiangya Hospital of Central South University between January and July 2024.Blood samples were tested using MetaCAP and conventional pathogen culture.The results of peritoneal fluid pathogen culture were used as the gold standard to compare the diagnostic efficacy of the two methods. Results:Due to three cases lacking peritoneal fluid culture results,a total of 26 cases were included in the final analysis.The overall mortality rate was 23.1％(6/26).During hospitalization,9 cases(34.6％)were diagnosed with IPN.The sensitivity and negative predictive value of MetaCAP for diagnosing IPN were significantly higher than those of conventional pathogen culture(77.8％vs.11.1％,P=0.031;86.7％vs.65.2％,P=0.032),while the differences in specificity(76.5％vs.88.2％,P=0.689)and positive predictive value(63.6％vs.33.3％,P=0.347)between the two methods were not statistically significant.The average detection time for MetaCAP was 33(20-49)h,while microbial culture took 125(45-142)h,with a significant difference(P＜0.001).The average cost for blood MetaCAP testing was 2 500 yuan per case,but it accounted for only 1.19％of the average hospitalization cost. Conclusion:MetaCAP has significant value in the early pathogen diagnosis of IPN,with a shorter detection time,good testing efficacy,and health-economic value,demonstrating a promising clinical application prospect.

Objective To investigate the effect of down-regulating adenosine deaminase acting on RNA-1(AD AR1)on the radiosensitivity of lung adenocarcinoma cells.Methods Lentiviral transfection was used to establish an ADAR1 knockdown cell line based on A549 cells.Then the cells were divided into negative control(shNC)and ADAR1 knockdown(shADAR1)groups,which were followed by a single-dose irradiation of 0 Gy and 6 Gy X-rays.Western blotting and RT-PCR were utilized to detect the expression of AD AR1 at protein and mRNA levels,respectively.CCK-8 assay,wound healing assay and Transwell migration assay were applied to measure cell proliferation and migration abilities.Meanwhile,clone formation assay was performed to detect the effect of down-regulating ADAR1 on the radiosensitivity of A549 cells.Flow cytometry and Western blotting were conducted to detect the expression levels of apoptosis and apoptosis-related proteins Bax and Bcl-2.Immunofluorescence assay and Western blotting were used to detect the expression level of γ-H2AX.Comet assay was performed to detect the level of cellular DNA damage.Twelve female nude mice(4～6 weeks old,weighing 16～18 g)were divided into shNC group,shADAR1 group,shNC+ionizing radiation(IR)group and shADAR1+IR group,with 3 mice in each group.The growth of tumor of different groups was observed with subcutaneous tumorigenesis assay.Results Western blotting and RT-qPCR showed that the protein and mRNA expression of ADAR1 were significantly reduced in A549 shADAR1 cells(P＜0.05).CCK-8 assay,wound healing assay and Transwell migration assay indicated that down-regulation of ADAR 1 inhibited the proliferation and migration abilities of A549 cells,and this inhibition trend became more obvious(P＜0.01)after IR.Cell clone formation assay showed that the clone formation rate of both groups was decreased,with the increase of radiation dose.But the number of formed clones was lower in the shADAR1 group than the shNC group.Flow cytometry and Western blotting displayed that down-regulation of AD AR1 increased the apoptotic rate and Bax expression in A549 cells(P＜0.01)and decreased Bcl-2 expression(P＜0.05),and the apoptotic rate and Bax protein level were further increased in A549 shADAR1 cells after IR(P＜0.01),and the Bcl-2 protein level was further decreased(P＜0.01).The number of γ-H2AX foci and protein level in A549 shADAR1 cells were significantly increased after IR(P＜0.05),and the results of comet assay showed that the DNA damage was more obvious in A549 shADAR1 cells after IR(P＜0.01).Subcutaneous tumorigenesis assay in nude mice showed that the growth of subcutaneous tumour of A549 shADAR1 cells was significantly inhibited after IR(P＜0.01).Conclusion Down-regulation of ADAR1 significantly inhibits the proliferation and migration of A549 cells after IR and promotes apoptosis and DNA damage,and thereby increases the radiosensitivity of lung adenocarcinoma cells.

Objective:To establish a method for the determination of 2, 3-Butanedione (BUT) in the air of workplace, which including the process of collection by absorption in phosphoric acid aqueous solution and the process of analysis and detection by high performance liquid chromatography with derivatization.Methods:In October 2022, a porous glass plate absorption tube containing 10 ml of 0.01% phosphoric acid solution was used to collect BUT in the air of the workplace at a flow rate of 0.2 L/min. The absorption solution was derived by 2, 4-dinitrophenylhydrazine for 75 min and separated on a SB-C18 column (250 mm×4.6 mm, 5 μm) . At the column temperature of 30 ℃, the mixture of acetonitrile-water ( V∶ V, 1∶1) was eluted at the flow rate of 1.0 ml/min. It was detected by UV detector (λ=365 nm) , qualitatived by retention time and quantitatived by external standard. Results:It showed that BUT in phosphoric acid aqueous solution could be stored for at least 7 d at 4 ℃. There was a linear relationship within the determination range of 0.05-6.00 μg/ml, the linear regression equation was y=89.610 x+0.133, r=0.9999. The sampling absorption efficiencies were 98.33%-100.00%, the detection limit of the method was 0.005 μg/ml, the minimum detection concentration was 0.016 mg/m 3 (based on V0=3.0 L) . The recovery rates were 95.96%-102.44%, the intra batch precision were 4.36%-7.78%, and the inter batch precision were 4.96%-6.06%. Conclusion:The method has the advantages of simple operation, high sensitivity and good accuracy. It can prevent the loss and degradation of BUT. It can be used for the determination of BUT in the air of workplace.

Objective:To establish a method for the determination of 2, 3-Butanedione (BUT) in the air of workplace, which including the process of collection by absorption in phosphoric acid aqueous solution and the process of analysis and detection by high performance liquid chromatography with derivatization.Methods:In October 2022, a porous glass plate absorption tube containing 10 ml of 0.01% phosphoric acid solution was used to collect BUT in the air of the workplace at a flow rate of 0.2 L/min. The absorption solution was derived by 2, 4-dinitrophenylhydrazine for 75 min and separated on a SB-C18 column (250 mm×4.6 mm, 5 μm) . At the column temperature of 30 ℃, the mixture of acetonitrile-water ( V∶ V, 1∶1) was eluted at the flow rate of 1.0 ml/min. It was detected by UV detector (λ=365 nm) , qualitatived by retention time and quantitatived by external standard. Results:It showed that BUT in phosphoric acid aqueous solution could be stored for at least 7 d at 4 ℃. There was a linear relationship within the determination range of 0.05-6.00 μg/ml, the linear regression equation was y=89.610 x+0.133, r=0.9999. The sampling absorption efficiencies were 98.33%-100.00%, the detection limit of the method was 0.005 μg/ml, the minimum detection concentration was 0.016 mg/m 3 (based on V0=3.0 L) . The recovery rates were 95.96%-102.44%, the intra batch precision were 4.36%-7.78%, and the inter batch precision were 4.96%-6.06%. Conclusion:The method has the advantages of simple operation, high sensitivity and good accuracy. It can prevent the loss and degradation of BUT. It can be used for the determination of BUT in the air of workplace.

【Objective】 To evaluate the application of monoclonal typing reagents and human anti-A/B antibodies for absorption-elution test in ABO grouping. 【Methods】 The specificity of monoclonal typing reagents and human anti-A/B antibodies with standard A, B, O and AB phenotypes at 4 ℃, room temperature, and 37 ℃ were compared. Affinity was evaluated by the titer, agglutination time and agglutination intensity of the reaction with A1/B cells. 29 samples with ABO discrepancy were tested to evaluate the ability of monoclonal typing reagents and human anti-A/B antibodies to detect weak antigens in absorption-elution test. 【Results】 The specificity and affinity of human anti-A/B antibodies are low, and monoclonal typing reagents have cross reactivity. Human anti-A/B antibodies can detect most weak antigens in absorption-elution test with no cross reactivity. 【Conclusion】 In ABO grouping, the human anti A/B antibody binding absorption-elution test can serve as a supplement method for identifying ABO weak antigens. Accurate results can be obtained with reasonable reagents and corresponding methodology in serological tests,thus ensuring the safety of blood transfusion.