To construct a recombinant expression system for a single-domain antibody targeting the urease of Helicobacter pylori(Hp),this study employed several strategies.First,using artificial intelligence(AI)auxiliary tools such as Pymol,I-TASSER,and ClussPro2,the molecular interactions between different antibodies and Hp urease subunit B(UreB)were analyzed.The fully humanized single-domain antibody UreBAb was identified as the primary research target.Next,the UreBAb gene sequence was optimized based on Escherichia coli codon preferences,and was inserted into expression vectors such as pET28a and pE-SUMO.The resulting recombinant expression strains were obtained by transforming Escherichia coli Rosetta(DE3).Recombinant antibody proteins were prepared through IPTG induction,and its activity was detected using extracted Hp urease as the antigen.SDS-PAGE analysis confirmed the correct expression of both UreBAb and SUMO-UreBAb,with protein yields of 0.34 mg/mL and 0.41 mg/mL,respectively.Unidirectional immunodiffusion experiments further confirmed that both recombinant antibodies exhibited strong affinity for Hp UreB antigen,with inhibition rates of 51.27%and 74.07%,respectively.Additionally,leveraging artificial intelligence tools such as AlphaFold2,cluspro2,mCSM-AB,OSPREY,and FoldX,the study evaluated and analyzed key binding sites and mutational strategies affecting the stability of the antigen-antibody complex.Subsequently,nine UreBAb evolution mutants were constructed,and their binding activities with the antigen were enhanced.Among these,the I107W mutant showed the most significant improvement,achieving a 24.95%increase compared to the wild-type UreBAb.This research lays a solid foundation for the development of fully humanized single-domain antibodies against Hp.

Objective To explore the effect of glioma stem cells with high expression of protein tyrosin phosphatase receptor type Z1 (PTPRZ1 )on the phenotypic polarization and phagocytosis of tumor-associated macrophages and its regulatory mechanism.Methods GSCs and non-stem tumor cells (NSTCs) were screened out from human glioblastoma (GBM) specimens using flow cytometry,and the PTPRZ1 expression in paired GSCs and NSTCs were detected.Human peripheral blood mononuclear cells (PBMC)-derived CD14+monocytes were exposed to the conditioned medium from glioma cells or recombinant chemokine C-C motif ligand 20 (CCL20)for TAM polarization.Stable PTPRZ1 knockout GSCs (PTPRZ1-KO GSCs) were constructed using CRISPR/Cas9. TAM phagocytosis to GSCs,NSTCs,PTPRZ1-Control GSCs (PTPRZ1-Ctrl GSCs)and PTPRZ1-KO GSCs and the expression of immunosuppressive phenotype (M2) polarization marker CD163 were examined using flow cytometry.Differentially expressed genes (DEGs ) between paired GSCs and NSTCs were determined using a bulk RNA-sequencing dataset (GSE54791 )from Gene Expression Omnibus (GEO).A gene set informing worse outcome of patients with GBM was generated using The Cancer Genome Atlas (TCGA)-GBM cohort.By intersecting the aforementioned gene set with the gene set that encodes for human membrance proteins,the PTPRZ1 gene is obtained.Gene set enrichment analysis (GSEA)was used for pathway enrichment analysis to compare the differentially regulated pathways between GBMs with high or low PTPRZ1 expression.Bulk RNA sequencing,qRT-PCR and Western blotting were used to identify the DEGs between PTPRZ1-KO GSCs and PTPRZ1-Ctrl GSCs.Results GSCs were more capable of escaping from TAM phagocytosis than NSTCs (P<0.05 )and had specifically up-regulated PTPRZ1 expression.PTPRZ1-KO significantly suppressed GSCs escaping from TAM phagocytosis (P<0.01 ). GBMs with high PTPRZ1 expression showed significant inhibition of pathways mediating phagocytosis (P<0.05).The expression of CCL20 as a M2 TAM polarization chemokine was significantly down-regulated in PTPRZ1-KO GSCs (P<0.05 ).Treatment with recombinant CCL20 up-regulated the expression of CD163 as a M2 TAM marker in TAM.Conclusion PTPRZ1+GSCs mediate M2 TAM polarization and inhibit TAM phagocytosis,which may be related to the up-regulation of CCL20 in PTPRZ1+GSCs.

Central venous catheterization plays an important role in the rescue of critically patients. Commonly used central veins in clinical practice include subclavian vein, internal jugular vein, and femoral vein. Serious complications after catheterization can endanger patients′ lives in severe cases. It is necessary to improve the understanding and treatment of serious complications of central vein catheterization in clinical work. A case of iliac vein dissection caused by right femoral vein catheterization was summarized in this article, and the catheter was successfully removed under digital subtraction angiography direct vision after the distal end of the catheter penetrated the vascular wall and reached the peritonea, which provided reference for the treatment of iatrogenic injury caused by central vein catheterization.

Objective To analyze the efficacy and effects of balloon dilation combined with iliac vein balloon dilatation on complications and coagulation function of acute deep venous thrombosis of lower limbs(DVT)complicated with Cockett syndrome.Methods Data of 102 patients with acute DVT combined with Cockett syndrome admitted to the hospital from March 2018 to September 2022 were retrospectively analyzed,including 53 patients treated with catheter thrombolytic balloon dilation of iliac vein(recorded as the study group)and 49 patients treated with catheter thrombolytic therapy(recorded as the control group).Both groups were followed up for 1 year.Thrombosis clearance,edema reduction of affected limb,lower limb hemodynamics,coagulation function,peripheral blood inflammatory factors,complications and thrombosis recurrence were compared between the two groups.Results The thrombus removal grade of the study group was better than that of the control group(P＜0.05).Before and 1 year after surgery,In the control group,the leg circumference diameter difference above the knee between the affected side and the healthy side was(8.02±1.41)cm and(2.89±0.32)cm,respectively.In the study group,the data were(8.19±1.38)cm and(2.57±0.29)cm,respectively.Before and 1 year after surgery,in the control group,the leg circumference diameter difference below the knee between the affected side and the healthy side was(6.84±1.18)cm and(2.13±0.38)cm,respectively.In the study group,the data were(6.63±1.09)cm and(1.76±0.32)cm,respectively.The difference was statistically significant(P＜0.05),and the study group was lower 1 year after surgery(P＜0.05).The average blood flow velocity of femoral vein in control group was(24.75±4.03)cm/s and(28.82±4.29)cm/s before and 3 days after surgery,respectively.In the study group,the data were(24.02±3.86)cm/s and(30.94±4.37)cm/s,respectively.The femoral vein blood flow before and 3 days after surgery in the control group was(13.02±2.12)ml/s and(15.05±2.29)ml/s,respectively,while that in the study group was(13.36±2.09)ml/s and(16.26±2.34)ml/s,with statistical significance(P＜0.05).The study group was higher 3 days after surgery(P＜0.05).The prothrombin time before and 3 days after operation were(22.93±2.04)s and(18.13±1.34)s in the control group,and(23.24±1.99)s and(17.29±1.21)s in the study group,respectively.The thrombin time before and 3 days after operation were(24.86±2.31)s and(21.04±1.75)s in the control group,and(24.13±2.16)s and(19.89±1.53)s in the observation group,respectively.The activated partial thromboplastin time before and 3 days after surgery was(59.21±3.92)s and(49.13±3.02)s in control group,and(60.17±3.85)s and(47.09±2.98)s in observation group,respectively,and the difference was statistically significant(P＜0.05).It was lower 3 days after surgery in study group(P＜0.05).In the study group,preoperative tumor necrosis factor(TNF)-α,platelet activating factor(PAF)and thromboxen B2(TXB2)were(31.91±4.89)ng/L,(14.59±2.36)pg/ml,and(213.12±30.98)pg/ml,respectively.Three days after surgery,the levels were(36.24±4.29)ng/L,(16.12±2.59)pg/ml,and(239.86±32.85)pg/ml,respectively,with statistical significance(P＜0.05).TNF-α,PAF and TXB2 were higher in the study group 3 days after surgery(P＜0.05).There was no significant difference in the incidence of total complications between the two groups(P＞0.05).The recurrence rate of thrombosis in study group was lower than that in control group(3.77％vs 16.33％,P＜0.05).Conclusion Catheter thrombolytic balloon dilation of iliac vein for acute DVT complicated with Cockett syndrome can enhance the thrombolysis effect,improve the swelling of the affected limb,lower limb hemodynamics and coagulation function,and reduce the risk of thrombosis recurrence,which is safe and reliable.However,this treatment plan can mediate the occurrence of inflammation,and reasonable anti-inflammatory therapy should be actively implemented after surgery.

Objective To investigate the effect of ultrasound-guided bilateral superior laryngeal nerve internal branch block combined with endotracheal surface anesthesia in patients with hypertensive cerebral hemorrhage for the duration of postoperative tube insertion.Methods A total of 100 emergency hypertensive intracerebral hemor-rhage surgical patients who visited our hospital from October 2021 to April 2023 were included as the research subjects.They were randomly divided into four groups,25 patients in each group.After surgery,group U received bilateral superior laryngeal nerve internal branch block under guidance of ultrasound combined with endotracheal surface anesthesia,group C1 received bilateral superior laryngeal nerve internal branch block under guidance of ultrasound,group C2 received endotracheal surface anesthesia,and group C3 did not undergo any procedure after surgery.Hemodynamic changes(HR,MAP,and SpO2)at different time points during the postoperative tube insertion in four groups of patients were recorded.The frequency of restlessness within 10 hours after surgery,the dosage of dexmedetomidine and urapidil,the volume of wound drainage,and the satisfaction of bed nurses were also recorded.Results There was no statistically significant difference in general conditions among the four groups(P>0.05).There was no statistically significant difference in HR and MAP at different time points in Group U(P>0.05),while the differences among the other three groups were statistically significant(P<0.05).At the same time point,the MAP and HR of group U and C1 were significantly lower than those of group C3(P<0.05),and the MAP of group U was significantly lower than that of group C2(P<0.05).There was no statistical difference in SPO2 among the four groups of patients at the same time point(P>0.05);The frequency of restlessness,dosage of dexmedetomidine and urapidil,and volume of wound drainage in the U and C1 groups were significantly lower than those in the C2 and C3 groups(P<0.05).Except for the difference in restlessness frequency(P<0.05),there was no statistical difference in other indicators between group U and C1;There was a statistical difference in satisfaction among the four groups of nurses(P<0.05,C3>C2>C1>U group).No nerve block related complications were observed in the U and C1 group.Conclusion Ultrasound-guided bilateral superior laryngeal nerve branch block combined with endotracheal surface anesthesia can maintain hemodynamics steadily of the postoperative patients in the NICU to varying degrees and reduce the frequency of postoperative restlessness,the dosage of sedative and anti-hypertensive drugs,while reducing the flow of wound drainage,and improve the satisfaction of bed nurses.

A 3-year-old male patient was diagnosed with neurofibromatosis type 1(NF1) for two years. The patient has multiple neurofibromas in retroperitoneum, lumbococcygeal paravertebral, lumbosacral spinal canal, and foramina. Due to retroperitoneal mass compression, the child suffered from urological complications such as hydronephrosis, ureterdilation, neurogenic bladder, etc., which seriously affected the urination function and resulted in multiple surgical treatments. Currently, the patient has been treated with mitogen activates extracelluar signal-regulated kinases(MEK) inhibitor selumetinib targeted therapy, and has voluntarily urinated, and his general state is better than before medication. The diagnosis and treatment of this case reflects the importance of multidisciplinary collaboration in the diagnosis and treatment of rare diseases.

Objective:To evaluate the efficacy and safety of endoscopic retrograde cholangiopancreatography (ERCP) after pancreaticoduodenectomy and endoscopic selection strategies.Methods:Clinical data of 34 patients treated with ERCP after pancreaticoduodenectomy at the Endoscopic Center of the First Affiliated Hospital of Air Force Medical University from January 2013 to December 2021 were retrospectively analyzed. The success rates of endoscopic insertion, diagnosis, treatment and ERCP, and the incidence of adverse events were analyzed.Results:Fifty ERCP treatments were performed in 34 patients. The success rates of endoscopic insertion, diagnosis, treatment, and ERCP after pancreaticoduodenectomy were 92.0% (46/50), 93.5% (43/46), 88.4% (38/43) and 76.0% (38/50), respectively. The success rates of ERCP assisted with colonoscope and balloon-assisted enterosocpe were 76.0% (19/25) and 75.0% (18/24), respectively. There were 3 adverse events, including 1 case of anastomotic mucosa tear during surgery, 1 case of cardiopulmonary arrest and 1 case of postoperative cholangitis.Conclusion:ERCP is effective and safe after pancreaticoduodenectomy in general. ERCP assisted with colonoscope and balloon-assisted colonoscope shows similar success rate after pancreaticoduodenectomy.

【Objective】 To explore the effects of insulin on the QT interval and induced arrhythmias of cardiomyocytes derived from human induced pluripotent stem cells （hiPSC-CMs）. 【Methods】 Immunofluorescence staining and flow cytometry were used to analyze the purity of hiPSC-CMs. Microelectrode array （MEA） was utilized to detect the electrophysiological changes including heart rate （HR）， field potential duration （FPD， which is similar to QT interval in ECG）， FPDc （FPD corrected by HR）， conduction velocity （CV）， and spike amplitude before and after insulin treatment. The effects of E4031 on QT interval prolongation and induced arrhythmias of hiPSC-CMs were evaluated before and after treatment with insulin. 【Results】 hiPSC-CMs highly expressed myocardial specific marker cTnT. The purity of hiPSC-CMs was 97.1%. After 5-day insulin treatment of hiPSC-CMs， HR increased by （11.9±3.3）%， FPD shortened by （22.7±2.8）%， FPDc shortened by （15.6±1.6）%， and spike amplitude increased by （39.1±7.9）% when compared with untreated group， but CV remained unchanged. 10 nmol/L of E4031 could prolong the FPDc of hiPSC-CMs by （37.8±9.0）%， and 30 nmol/L of E4031 could induce arrhythmias. After insulin treatment， 10 nmol/L of E4031 prolonged the FPDc of hiPSC-CMs by （21.8±3.1）% （compared with the untreated group， insulin decreased FPDc prolongation by E4031， 37.8%±9.0% vs. 21.8%±3.1%， P<0.05）， while 30 nmol/L of E4031 did not induce arrhythmias. 【Conclusion】 Insulin can shorten the QT interval of hiPSC-CMs and significantly reduce the QT interval prolongation and the risk of arrhythmias induced by drugs.

【Objective】 To study the clinical practice and early outcome of percutaneous full-endoscopic modified posterior lumbar interbody fusion（mPLIF）combined with pedicle screw fixation through paraspinal muscle clearance. 【Methods】 A retrospective study was conducted to analyze the clinical data of patients with lower lumbar spinal diseases treated from May 2019 to April 2020. All the enrolled patients received mPLIF combined with pedicle screw fixation through paraspinal muscle clearance. The follow-up period was more than 1 year； the general parameters included age， gender， duration of disease， diagnosis of disease， surgery segment， and postoperative hospitalization time. Operation parameters included operation time and blood loss. We obtained the clinical parameters such as visual analogue scale （VAS） score for back and lower extremity， Oswestry disability index （ODI） score， and Macnab satisfaction score at the last follow-up. We evaluated the imaging parameters including intervertebral disc height， segmental lordosis angle， lumbar lordosis angle， as well as fusion outcome of patients with single segmental lumbar disease. In addition， intraoperative and postoperative complications were recorded. 【Results】 Totally 18 patients met the inclusion criteria， among whom 8 were male and 10 were female， with the average age of （53.3±8.3） years old and the average duration of disease being （28.9±36.6） months. Among them 16 patients were diagnosed as lumbar degenerative disease and the other 2 had lumbar disc infection. One patient received L3-L4 and L4-L5 intervertebral fusion； the others had one-segmental fusion， among which 11 cases were L4-L5 and 6 cases were L5-S1. The average operation time was （207.8±31.7） min， and the average blood loss was （25.6±7.8） mL， and the mean postoperative hospitalization time was （6.56±2.30） days. VAS scores of back and lower extremities at postoperative 1 week， 6 months and the last follow-up were statistically significantly improved from the preoperative scores. ODI scores at postoperative 6 months and the last follow-up were also statistically significantly improved. The rate of excellent and good according to the Macnab criteria was 94.4%. For the 17 single-level fusion patients， intervertebral height was significantly higher postoperatively and at the last follow-up compared with that of the preoperative one （P<0.05）. Segmental lordosis angle was bigger postoperatively and at the last follow-up （P>0.05）， which was not statistically significant. Lumbar lordosis angle was significantly bigger postoperatively （P<0.05） and bigger at the last follow-up， but with no statistical significance （P>0.05）. The fusion rate at the last follow-up was 88.2%. The cage broke in the process of implantation in one patient. A cage retroposition occurred in one patient at the follow-up. 【Conclusion】 Percutaneous full-endoscopic modified posterior lumbar interbody fusion combined with pedicle screw fixation through bilateral paraspinal muscle clearance by one incision showed excellent clinical outcomes in treating many kinds of lower lumbar diseases. This operation should be an excellent option for lower lumbar fusion.

Extracellular vesicles (EVs), also known as membrane vesicles, are vesicular bodies secreted by eukaryotic cells and bacteria. EVs can carry proteins, DNA, RNA, and various metabolites for the exchange and transmission of substances between cells. They play contents-dependent physiological functions, such as delivering nutrients, participating in immune response, and treating cancers. Currently, most studies focus on the exploration of vesicles secreted by eukaryotic cells and gram-negative bacteria, while few studies focus on gram-positive bacteria. This review summarized the production, content composition, physiological function, and engineering of EVs secreted by gram-positive bacteria, and prospected future perspectives in this area.
Bacteria/metabolism* ; Extracellular Vesicles/metabolism* ; Gram-Negative Bacteria ; Gram-Positive Bacteria/metabolism* ; Proteins/metabolism*