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.

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 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.

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 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.

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 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.

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*

Objective:To establish surgical difficulty scoring system of partial nephrectomy based on holographic imaging and explore its application value in partial nephrectomy.Methods:A total of 184 patients including 110 males and 74 females with renal tumors diagnosed as stage cT 1 to cT 2 before surgery in the First Affiliated Hospital of Xiamen University from October 2019 to January 2022 were included. Among the 184 patients, 141 patients were treated with partial nephrectomy and 43 patients were treated with radical nephrectomy (3 partial nephrectomies were finally changed to radical nephrectomies due to vascular and tumor location). 60 patients had hypertension. 24 patients had diabetes. 7 patients had hyperuricemia. The median age was 55(47, 62) years. The median BMI was 23.7(21.8, 26.4) kg/m 2. The median maximum tumor diameter was 3.9(2.9, 5.2) cm. The median preoperative eGFR was 99.7(83.4, 114.2) ml/(min·1.73m 2). The median R. E.N.A.L. score was 8(6, 9). The median PADUA score was 9(8, 10). 153 patients were diagnosed as stage cT 1 and 31 patients were diagnosed as stage cT 2. The hologram reconstruction was performed according to preoperative CT or MRI examination. The maximum diameter of the tumor in the kidney (D), the compression degree of the renal segmental vessels by tumor(C), the area of the renal sinus occupied by tumor(O) and the mass of exophytic rate(M) were comprehensively considered and finally constituted the difficulty scoring system named DCOM score for partial nephrectomy. The DCOM score divided the complexity of tumor surgery into mild (4-6 points), moderate (7-8 points) and high (≥ 9 points). Meanwhile, the MIC (surgical margins are negative, WIT is <20 min, and no major complications)was used to evaluate the overall surgical effect. The DCOM, R. E.N.A.L. and PADUA scores were performed on all patients and compared with each other to evaluate the surgical effect of DCOM score in partial nephrectomy. Results:All surgeries in this study were successfully completed, including 141 partial nephrectomies and 43 radical nephrectomies. The DCOM score was 10(9, 11) for radical nephrectomy and 6(5, 8) for partial nephrectomy, and the difference was statistically significant ( P=0.001). There were 23 patients (37.7%) in highly complex group, 39 patients (88.6%) in moderately complex group and 79 patients (100.0%) in mildly complex group underwent partial nephrectomy, respectively. According to multifactorial analysis, patients in highly and moderately complex group of DCOM score had 8.88 times ( P=0.001) and 1.76 times ( P=0.005) less reach MIC than those in mildly complex group, respectively. Patients in highly and moderately complex group of PADUA score had 4.86 times ( P=0.005)and 3.41 times ( P=0.006)less reach MIC than patients in mildly complex group of DCOM score, respectively. What’s more, patients in moderately complex group of R. E.N.A.L. score had 3.11 times ( P=0.003) less reach MIC than patients in mildly complex group of DCOM score. In the ROC curves to predict MIC achievement, the AUC values of R. E.N.A.L., PADUA and DCOM scores were 0.657, 0.655 and 0.746, respectively. Comparing:R. E.N.A.L. score with DCOM score, the AUC value was statistically significant ( P=0.025). Conclusions The surgical difficulty scoring system (DCOM score) based on holographic imaging can predict the outcome of partial nephrectomy, but further verification is needed.

Objective:To evaluate the value of conventional ultrasound, shear wave elastic parameters and immunohistochemistry in predicting axillary lymph node metastasis of breast cancer.Methods:The ultrasonographic features and pathological results of 172 masses in 152 breast cancer patients who underwent surgery in the First Affiliated Hospital of Shihezi University Medical College from May to October 2020 were analyzed retrospectively. The patients were divided into metastatic group and non-metastatic group according to the status of axillary lymph nodes. The conventional ultrasound characteristics, shear wave velocity (SWV) and immunohistochemical indexes (ER, PR, HER-2, Ki-67) of 2 groups of breast cancer masses were analyzed. Finally, the parameters with statistically significant difference between groups were selected and the Logistic regression model was established.Results:There were significant differences in the aspect ratio, calcification, SWVmean and HER-2 expression between metastatic group and non-metastatic group (all P<0.05). A prediction model was constructed with aspect ratio >1, calcification, high SWVmean and HER-2(+ ). The area under receiver operating characteristic curve (AUC) of the subjects was 0.891, which was larger than the single parameter (all P<0.05), and was in good agreement with pathological results (Kappa=0.731). Conclusions:The joint prediction model can be used to predict the status of lymph nodes, and the axillary lymph node metastasis is more likely to occur in breast cancer with the aspect ratio >1, calcification, high SWVmean and HER-2(+ ).