Objective To develop an erythrocyte-based delivery system for butyrylcholinesterase(BChE)that is capable of prophylaxis against organophosphorus nerve agents.Methods Recombinant BChE was produced and analyzed for oligomerization via polyacrylamide gel electrophoresis(PAGE)and Western blotting.A modified hypotonic preswelling method was employed to prepare BChE-loaded erythrocytes.The drug loading capacity and encapsulation efficiency were quantified using enzyme-linked immunosorbent assay(ELISA).Catalytic activity was assessed in vitro with an activity detection kit.The system was characterized via scanning electron microscopy(SEM),flow cytometry and a hematology analyzer.Results Recombinant BChE predominantly existed as dimers(85％dimer,15％monomer).The optimized volume ratio of erythrocytes to hypotonic solution was determined as 1:7.Compared with native and empty erythrocytes,BChE-loaded erythrocytes exhibited significantly higher catalytic activity(P＜0.001).The mean corpuscular volume of BChE-loaded erythrocytes increased(P＜0.001),while the mean content of corpuscular hemoglobin and hemoglobin in erythrocytes per 100 mL decreased(P＜0.001).SEM revealed no morphological differences(biconcave disc shape).Hypotonic preswelling moderately increased erythrocyte apoptosis(P＜0.001),but no statistical difference was observed between BChE-loaded and hypotonic-treated erythrocytes(P＞0.05).CD47 expression remained unchanged compared to native erythrocytes(P＞0.05).Conclusion The modified hypotonic preswelling method can generate BChE-loaded erythrocytes that retain the characteristics of native erythrocytes while conferring catalytic activity,offering a novel strategy for clinical intervention against organophosphorus poisoning.

Objective To compare the clinical efficacy of laparoscopic high ligation of the hernia sac combined with intraperitoneal onlay mesh(IPOM)and local anesthesia Lichtenstein repair for the treatment of inguinal hernia after total cystectomy in male patients.Methods A total of 59 male patients who underwent total cystectomy were included in this study and randomly divided into two groups:the laparoscopic high ligation of the hernia sac combined with IPOM group(IPOM group,30 cases)and the local anesthesia Lichtenstein repair group(local anesthesia group,29 cases).The patient age,surgical time,postoperative ambulation time,visual analogue scale score for pain on the 1st to 3rd days and 3 months after operation,time to first mobilization post-operation,pain scores on postoperative day 1,2 and 3,pain scores at 3 months post-operation were observed and compared between the two groups.Data of intraoperative blood loss,hospital stay,time to return to daily activities,total surgery costs,recurrence rates at 1 year after operation and complications were also observed and compared in two groups of patients.The results were analyzed to determine the optimal surgical method.Results The IPOM group demonstrated better outcomes in terms of surgical time,time to first mobilization,pain scores on postoperative days 1,2 and 3,pain scores at 3 months,intraoperative blood loss,hospital stay and time to return to daily activities compared to those of the local anesthesia group(P＜0.01).However,the total cost of the operation was higher in the IPOM group than those of the local anesthesia group(P＜0.01).There were no significant differences in postoperative complications between the two groups(P＞0.05).The recurrence rate of the IPOM group one year after surgery was better than that of the local anesthesia group(P＜0.05).Conclusion Laparoscopic IPOM with high ligation of the hernia sac is superior to the traditional Lichtenstein repair in reducing postoperative pain and recurrence rates.This surgical method should be the first choice when economic conditions permit.

Objective To compare the clinical efficacy of laparoscopic high ligation of the hernia sac combined with intraperitoneal onlay mesh(IPOM)and local anesthesia Lichtenstein repair for the treatment of inguinal hernia after total cystectomy in male patients.Methods A total of 59 male patients who underwent total cystectomy were included in this study and randomly divided into two groups:the laparoscopic high ligation of the hernia sac combined with IPOM group(IPOM group,30 cases)and the local anesthesia Lichtenstein repair group(local anesthesia group,29 cases).The patient age,surgical time,postoperative ambulation time,visual analogue scale score for pain on the 1st to 3rd days and 3 months after operation,time to first mobilization post-operation,pain scores on postoperative day 1,2 and 3,pain scores at 3 months post-operation were observed and compared between the two groups.Data of intraoperative blood loss,hospital stay,time to return to daily activities,total surgery costs,recurrence rates at 1 year after operation and complications were also observed and compared in two groups of patients.The results were analyzed to determine the optimal surgical method.Results The IPOM group demonstrated better outcomes in terms of surgical time,time to first mobilization,pain scores on postoperative days 1,2 and 3,pain scores at 3 months,intraoperative blood loss,hospital stay and time to return to daily activities compared to those of the local anesthesia group(P＜0.01).However,the total cost of the operation was higher in the IPOM group than those of the local anesthesia group(P＜0.01).There were no significant differences in postoperative complications between the two groups(P＞0.05).The recurrence rate of the IPOM group one year after surgery was better than that of the local anesthesia group(P＜0.05).Conclusion Laparoscopic IPOM with high ligation of the hernia sac is superior to the traditional Lichtenstein repair in reducing postoperative pain and recurrence rates.This surgical method should be the first choice when economic conditions permit.

Objective To investigate the expression and clinical significance of LINC00638 in colon cancer,and further to explore the impact of LINC00638 on the biological function of colon cancer cells.Methods The cancer genome atlas(TCGA)database was used to preliminarily compare the expression differences of LINC00638 in normal colon tissue and colon cancer tissue,and to explore the rela-tionship between LINC00638 and clinical pathological characteristics of patients.Kaplan-Meier survival analysis was used to further in-vestigate the effect of LINC00638 expression on the prognosis of colon cancer patients.The GEPIA database and the Gene Expression Om-nibus(GEO)database were used to further validate the effect of LINC00638 expression on the prognosis of colon cancer patients.Univari-ate and multivariate COX regression analysis were used to explore the effect of LINC00638,clinical and pathological related factors on the prognosis of colon cancer patients.Small interfering RNA(siRNA)was used to silence the expression of LINC00638 in HCT116 cells and DLD1 cells,and the effect of LINC00638 on the proliferation,migration and invasion of colon cancer cell was explored through CCK-8 experiment,colony formation experiment,scratch experiment and Transwell experiment.Results The expression level of LINC00638 was significantly increased in colon cancer tissue,and its expression level was closely related to lymph node metastasis and TNM staging in co-lon cancer patients.The results of Kaplan-Meier survival curve analysis showed that the higher the expression level of LINC00638,the worse the prognosis of colon cancer patients(P＜0.05).The results of COX regression analysis showed that LINC00638,age,and TNM staging could be independent prognostic factors for colon cancer patients.The results of cell function experiments showed that compared with the control group,the proliferation,migration,and invasion ability of cells in knockdown group were significantly decreased(P＜0.05).Conclusion LINC00638 is highly expressed in colon cancer,and the higher its expression level,the worse the prognosis of colon cancer patients.After knocking down LINC00638,it can inhibit the proliferation,migration and invasion ability of colon cancer cells.

Objective To investigate the pathological damage to and inflammation of different intestinal segments in a rat model of severe hemorrhage,and to explore the effect of polarization of intestinal macrophage on the pathophysiology of intestinal inflammation.Methods Male Wistar rats were randomly divided into two groups:the sham operation group and hemorrhage group.In the hemorrhage group,40％of the total blood volume was lost in 25-30 minutes,while in the sham operation group,only the femoral artery and vein were intubated without bleeding.The rats were killed at 0,3,6,12 and 24 hours.The entire intestine was isolated quickly,and sections of the intestine were cut at the duodenum,jejunum,ileocecal junction,colon and rectum for histopathological evaluation.ELISA was adopted to determine related inflammation factors while multi-color immunohistochemistry was used to calculate macrophage surface markers.The data was statistically analyzed.Results(1)Compared with the sham group,there was no significant difference in colon histology at 3 h and 6 h,but significant difference was detected in rectum scores only at 24 h.The scores of other intestinal segments were significantly different at each time point.The severity of ileocecal and colonic lesions after bleeding increased with time.The duodenum,jejunum and ileocecum were more critically injured at 3 h than the rectum at 6 h.The injury to the duodenum,jejunum,ileum and colon was much more pronounced than to the rectum at 12 h.(2)The expressions of TNF-α and IL-1β in the rectum were increased significantly at 12 h post operation.The expressions of IL-1β,TNF-α in the jejunum increased obviously at 3 h and 6 h,respectively.(3)Three hours after severe bleeding,the level of macrophages in the jejunum and ileocececal area increased significantly,and the percentage of M1 macrophages was higher.After 6 hours,the proportion of M2 macrophages in the jejunum and M1 macrophages decreased significantly.After 3 hours,the percentage of M1 macrophages in the colon decreased,but that of M2 macrophages increased.The proportion of M2 polarized macrophages in the duodenum and rectum increased at 3 h after severe bleeding but decreased at 6 h.Conclusion Pathological damage to intestinal sections after bleeding varies depending on the time,and is correlated with the inflammatory level of macrophages.

ObjectiveTo systematically evaluate the efficacy and safety of omental wrapping technique for pancreaticojejunal anastomosis in preventing complications after pancreaticoduodenectomy. MethodsThis study was conducted according to the PRISMA guideline. English and Chinese databases including CNKI， Wanfang Data， VIP， CBM， the Cochrane Library， PubMed， Embase， and Web of Science were searched for clinical studies on omental wrapping technique for pancreaticojejunal anastomosis in preventing complications after pancreaticoduodenectomy published up to November 2022， and Stata 16 and Review Manager 5.4 were used to perform the meta-analysis. ResultsA total of 15 studies with 1 830 patients were included in this study. The meta-analysis showed that the omental wrapping group had a significantly lower overall incidence rate of postoperative pancreatic fistula （POPF） than the non-omental wrapping group （odds ratio ［OR］=0.30， 95% confidence interval ［CI］： 0.22‍ ‍—‍ ‍0.41， P<0.001）， and the subgroup analysis showed that the omental wrapping group had a significantly lower incidence rate of grade B/C POPF than the non-omental wrapping group （OR=0.29， 95%CI： 0.21‍ ‍—‍ ‍0.39， P<0.001）. Compared with the non-omental wrapping group， the omental wrapping group had significantly lower incidence rates of postoperative bile leakage （OR=0.30， 95%CI： 0.16‍ ‍—‍ ‍0.56， P<0.001）， postoperative hemorrhage （OR=0.35， 95%CI： 0.24‍ ‍—‍ ‍0.53， P<0.001）， delayed gastric emptying （OR=0.45， 95%CI： 0.31‍ ‍—‍ ‍0.64， P<0.001）， abdominal infection （OR=0.55， 95%CI： 0.40‍ ‍—‍ ‍0.75， P<0.001）， reoperation （OR=0.31， 95%CI： 0.18‍ ‍—‍ ‍0.54， P<0.001）， and death within 30 days after surgery （OR=0.42， 95%CI： 0.22‍ ‍—‍ ‍0.80， P=0.009）， a significantly earlier time to diet （mean difference ［MD］=-0.98， 95%CI： -1.84 to -0.11， P=0.03）， and a significantly shorter length of postoperative hospital stay （MD=-2.44， 95%CI： -4.10 to -0.77， P=0.004）. There were no significant differences between the two groups in the time of operation （MD=-13.68， 95%CI： -28.31 to -0.95， P=0.07） and intraoperative blood loss （MD=-17.26， 95%CI： -57.55 to -23.03， P=0.40）. ConclusionOmental wrapping can reduce the incidence rates of postoperative complications such as pancreatic fistula， bile leakage， postoperative hemorrhage， abdominal infection， and delayed gastric emptying， improve the prognosis of patients， and shorten the length of hospital stay， without increasing surgical difficulty or time of operation.

Objective Based on the gene expression omnibus(GEO)database,bioinformatics methods were employed to analyze the expression characteristics of hypoxia-related differentially expressed genes(HRDEGs)in ischemic stroke,and key genes were screened,to provide important support for a deeper understanding of ischemic stroke.Methods The GSE16561 and GSE58294 datasets were downloaded from the GEO database,and Python software was used for data integration.The Combat method was employed to eliminate batch effects while retaining disease grouping characteristics.Principal component analysis was conducted to reduce dimensionality of the data before and after batch effect removal,and intraclass correlation coefficient(ICC)testing was performed on the ischemic stroke and normal control groups.Gene set enrichment analysis(GSEA)and single-sample GSEA were conducted on the merged and batch effects eliminated dataset,with a nominal P-value(NOM P-val)＜0.05 and false discovery rate P-value(FDR P-val)＜0.25 used as criteria to select significantly different gene sets.Differential expression genes between the ischemic stroke samples and normal control samples after merging and eliminating batch effects of the GSE16561 and GSE58294 datasets were identified using R software,with an absolute value of log2 gene expression fold change(FC)≥0.58 and adjusted P-value(Padj)＜0.05 as selection criteria.Intersection with hypoxia-related genes obtained from the National Center for Biotechnology Information(NCBI)in the United States yielded the HRDEGs.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were performed on the HRDEGs,and the STRING database was used to construct a protein-protein interaction network of differentially expressed genes.The top 10 key genes were filtered using Cytoscape 3.8 software.Results The ICC analysis results showed excellent consistency in the ischemic stroke and normal control samples after batch effect removal,with ICC values of 0.94 and 0.98 for the GSE16561 and GSE58294datasets,respectively.GSEA results demonstrated significant enrichment of 34 gene sets in the stroke samples in the newly merged and batch effects removed dataset from GSE16561 and GSE58294,leading to the identification of 404 differentially expressed genes(all with Padj＜0.05),including 354 upregulated genes and 50 downregulated genes.Intersection with hypoxia-related genes yielded 64 HRDEGs.GO enrichment analysis indicated significant enrichment of HRDEGs in vesicle lumen,cytoplasmic vesicle lumen,secretory granule lumen,with molecular functions such as amide binding,peptide binding,phospholipid binding,and enzyme inhibitor activity.These genes are primarily involved in the positive regulation of cytokine production,regulation of immune response,response to bacterium-derived molecules,and response to lipopolysaccharide,among other biological processes.KEGG enrichment analysis revealed enrichment of HRDEGs in pathways related to lipid and atherosclerosis,Salmonella infection,neutrophil extracellular trap formation,nucleotide-binding oligomerization domain-like receptor signaling pathway,protein glycosylation in cancer,tuberculosis,and necroptosis.Based on the protein-protein interaction network,10 key genes were identified,including arginase1(ARG1),caspase1(CASP1),interleukin1 receptor type 1(IL-1R1),integrin subunit alpha M(ITGAM),matrix metalloproteinase9(MMP9),prostaglandin-endoperoxide synthase 2(PTGS2),signal transducer and activator of transcription 3(STAT3),Toll-like receptor2(TLR2),TLR4,and TLR8.Conclusion This study has identified 10 key genes associated with ischemic stroke and hypoxia through bioinformatics mining,which maybe provid potential targets for subsequent research and diagnostic and therapeutic interventions.

Objective:To investigate the unqualified hepatitis C virus (HCV) detection result of blood donors from the served area of blood institutions.Methods:The data related to HCV markers detected of the first and repeat blood donors were collected from the system of practice comparison for the Chinese mainland blood institutions from 2017 to 2021. The anti-HCV reactive rate and the rates of anti-HCV negative but HCV-RNA reaction and all the relationship between rates and the annual, regional and different blood donors were statistically analyzed.Results:During 2017-2021, the number of anti-HCV reactive per 100 000 blood donors decreased from 444.3 to 250.44 in the served area of 22 blood institutions ( χ2=49.677, P<0.05). The number of HCV RNA detected positive per 100 000 anti-HCV negative increased from 0.69 to 2.05 year by year, but there was no statistical significance ( χ2=0.643, P>0.05). The anti-HCV unqualified rate was significantly different among regions ( χ2=3 260.283, P<0.05). The anti-HCV unqualified rate of the first blood donors was significantly higher than that of the repeated blood donors ( F=130.993, P < 0.05). The annual number of HCV RNA detected positive per 100 000 anti-HCV negative blood samples from donors ranged from 0 to 17.28. Conclusions:The anti-HCV unqualified rate of blood donors in the served area of 22 blood institutions decreased year by year. Compared with repeated blood donors, HCV infection should be emphasized in first-time blood donors. The implementation of HCV RNA test can detect out much more HCV infections and reduce the risk of transfusion transmitted infectious HCV.

Objective:To analyze the clinical characteristics and pathogenic distribution of severe pneumonia in adults in order to provide basis for clinical diagnosis and treatment.Methods:From June 2021 to April 2022, 145 patients with pneumonia admitted to the Department of Respiratory and Critical Care Medicine of the Second People's Hospital of Guangdong Province. According to whether they meet the diagnostic criteria for severe pneumonia, they were divided into severe ( n=63) and mild ( n=82) groups, and the clinical features between the two groups were compared. At the same time, the role of FilmArray detection in severe pneumonia was discussed. The measurement data were tested using independent sample t test or Mann-Whitney U test, and the counting data were tested using Chi-square test or Fisher exact probability method. Results:The age of the patients in the severe group was (72.67±1.71) years, male patients accounted for 84.1%, and the median hospitalization time was 16 days. Nine patients died in hospital; most of them had fever, shortness of breath, and change of consciousness, accompanied by hypertension, diabetes, cerebrovascular disease, chronic kidney disease, and tumor history. Compared with the mild group, the total number of leukocytes, neutrophil ratio, procalcitonin, and C-reactive protein were higher in the severe group, but the CD3 +, CD4 +, and CD8 + cell counts were lower ( P<0.05). The positive rate of FilmArray detection in the severe group was 81%, and the mixed infection of multiple bacteria accounted for 50%, which was higher than that of traditional culture ( P<0.05). The top four pathogens in severe group were Pseudomonas aeruginosa, Acinetobacter baumannii complex, Klebsiella pneumoniae, and Staphylococcus aureus, which were significantly higher than that in the mild group ( P<0.05). Resistance genes were detected in patients with severe disease, which was significantly higher than that in patients with mild disease (70.7% vs. 17.5%, P<0.05). Conclusions:Severe pneumonia is more common in elderly men, with more basic diseases and poor immunity. FilmArray has a high positive rate and can detect multiple pathogens, which may have a role in the rapid diagnosis of severe pneumonia.

With the promotion of the concept of minimally invasive surgery, hepatobiliary surgery has gradually become a mainstream technique; however, for bile duct tumors in the perihilar region, there are few reports on the application of laparoscopy in such tumor due to the complex structure and anatomical variations of the hepatic hilar region. With reference to related reports in China and globally, this article introduces the research advances in the application of laparoscopy in perihilar tumor surgery. The analysis shows that laparoscopy has certain advantages in the surgical treatment of perihilar tumors, especially in the treatment of Bismuth Ⅰ/Ⅱ hilar cholangiocarcinoma and early-stage gallbladder cancer; due to the high requirements of laparoscopic surgery for the surgeon, only a few experts can perform this surgery at present, and strict screening and evaluation of patients are also required. Therefore, laparoscopy used in perihilar tumor surgery is still at the preliminary stage of exploration, with a need for further exploration and learning.