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 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.

To investigate the effects of the S-curve leakage testing method on the detection of flexible endoscope and occurrence of medium to high-level faults of flexible endoscope. A convenience sampling method was used to study the information of endoscopic leak detection at the digestive endoscopic centre of the First Affiliated Hospital of Air Force Medical University. From July 2016 to December 2017, 58 endoscopes which received conventional leakage testing were set as the control group (29 116 tests). From January 2018 to June 2019, 56 endoscopes which received S-curve leakage testing were set as the observation group (28 112 tests). The results showed that the detection proportion of angular abnormalities in the observation group was higher than that in the control group [65.6% (59/90) VS 40.4% (36/89), χ2=11.330, P<0.001]. However, the detection proportion of medium to high-grade faults such as damaged charge coupled device (CCD) components and broken insertion section in the observation group was lower than that in the control group [0.04% (10/28 112) VS 0.08% (23/29 116), χ2=4.680, P=0.030]. Moreover, the overall maintenance cost was lower in the observation group than that in the control group (313.7 thousand yuan VS 942.6 thousand yuan). It is indicated that the S-curve leakage testing method can detect low-level endoscopic faults early such as angular abnormalities, and reduce the occurrence of medium to high-level endoscopic faults, which contributes to cost reduction and efficiency increase.

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.

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*

The enterobacterial common antigen (ECA) is a polysaccharide composed of polysaccharide repeats that are located in the outer membrane of almost all Enterobacteriaceae bacteria and has diverse biological functions. ECA is synthesized by the synergistic action of multiple genes that are present in clusters on the genome of Enterobacteriaceae bacteria, forming the ECA antigen gene cluster, an important virulence factor that plays a role in host invasion and survival of Enterobacteriaceae in vivo. ECA also plays an important role in the maintenance of the bacterial outer membrane permeability barrier, flagella gene expression, swarming motility, and bile salts resistance. In addition, ECALPS, anchored in the core region of bacterial lipopolysaccharide, is an important surface antigen for bacteria, stimulating high levels of antibody production in the host and could be a target for vaccine research. This review summarizes ECA purification, genes involved in ECA biosynthesis, its immunological characteristics, biological functions and clinical applications.
Antigens, Bacterial/genetics* ; Enterobacteriaceae/genetics* ; Lipopolysaccharides ; Polysaccharides

Deepening the ideological and political construction of curriculum and carrying out the fundamental task of cultivating people with morality are the important requirements of education reform and talent cultivation in the new era. Microbiology Experiment is an important basic course and core practice course of Bioengineering, Pharmaceutical Engineering, Food Science and Engineering, et al. In order to give full play to the education function of Microbiology Experiment, this article deeply developed the ideological elements contained in the curriculum referring to the guidelines for the construction of ideological and political courses in institutions of higher education. And the article explored the ideological and political reform of Microbiology Experiment from three aspects: teaching content reform, teaching method innovation and improvement of teachers' ideological and political construction ability. Strive to integrate the value shaping, knowledge transference and ability training, cultivate high-quality professionals with firm ideals and beliefs.
Curriculum ; Humans ; Universities

Objective:To explore the efficacy of endoscopic retrograde cholangiopancreatography (ERCP) biliary stent implantation in patients with unresectable malignant biliary stricture (MBS) and the influencing factors of overall survival.Methods:The clinical data of 346 patients who underwent ERCP biliary stent implantation due to MBS from May 2013 to October 2016 in Xijing Digestive Disease Hospital of Air Force Military Medical University, Shanxi Bethune Hospital and Mengchao Hepatobiliary Hospital of Fujian Medical University were retrospectively analyzed, and the efficacy, complications and risk factors affecting overall survival were also analyzed.Results:After ERCP biliary stent implantation, the levels of total bilirubin, γ-glutamyl transpeptidase, alkaline phosphatase and alanine aminotransferase were lower than those before surgery (all P < 0.01). The incidence of infection after operation was 14.7% (51/346), and the incidence of biliary infection was 13.0% (45/346). The incidence of post-ERCP pancreatitis (PEP) was 4.6% (16/346). The median survival time after ERCP was 131.0 d (70.3 d, 246.5 d). Multivariate Cox regression analysis showed that the independent risk factors affecting the overall survival patients included the hilar bile duct stenosis ( HR = 1.85, 95% CI 1.44-2.38, P < 0.01), preoperative bilirubin level exceeding the upper limit of normal level by 5 times ( HR = 1.75, 95% CI 1.30-2.36, P < 0.01), carbohydrate antigen 199 level exceeding the upper limit of normal level by 10 times ( HR = 1.27, 95% CI 1.00-1.61, P = 0.050), vascular and organ metastasis ( HR = 1.32, 95% CI 1.04-1.69, P = 0.023), and the poor jaundice decreasing level ( HR = 1.37, 95% CI 1.02-1.85, P = 0.037) . Conclusions:The ERCP biliary stent implantation is a safe and effective therapy for MBS. ERCP biliary stent implantation MBS patients with hilar bile duct stenosis, preoperative bilirubin levels more than 5 times of the upper limit of normal level, carbohydrate antigen 199 levels more than 10 times of the upper limit of normal level, vascular and organ metastasis, and poor jaundice decreasing level may have poor overall survival.

OBJECTIVE: To investigate the effects of millimeter wave (MMW) exposure on apoptosis of human melanoma A375 cells and explore the mechanisms. METHODS: Through electromagnetic field calculation we simulated MMW exposure in cells and calculated the specific absorption rate (SAR). The optimal irradiation parameters were determined according to the uniformity and intensity of the SAR. A375 cells were then exposed to MMV for 15, 30, 60, or 90 min, with or without pretreatment with the caspase-3 inhibitor AC-DEVD-fmk (10 μmol/L) for 1 h at 90 min before the exposure. CCK-8 assay was used to assess the changes in the viability and Annexin-V/ PI staining was used to detect the apoptosis of the cells following the exposures; Western blotting was used to detect the expression of caspase-3 in the cells. RESULTS: The results of electromagnetic field calculation showed that for optimal MMV exposure, the incident field needed to be perpendicular to the bottom of the plastic Petri dish with the antenna placed below the dish. CCk-8 assay showed that MMW exposure significantly inhibited the cell viability in a time-dependent manner ( < 0.05); exposures for 15, 30, 60, and 90 min all resulted in significantly increased apoptosis of the cells ( < 0.05). The cells with MMW exposure showed significantly increased expression of caspase-3. The inhibitory effect of MMW on the cell viability was antagonized significantly by pretreatment of the cells with AC-DEVD-fmk ( < 0.05), which increased the cell viability rate from (36.7±0.09)% to (59.8±0.06)% ( < 0.05). CONCLUSIONS 35.2 GHz millimeter wave irradiation induces apoptosis in A375 cells by activating the caspase-3 protein.
Apoptosis ; Caspase 3 ; metabolism ; Caspase Inhibitors ; pharmacology ; Cell Line, Tumor ; Cell Survival ; Electromagnetic Fields ; Enzyme Activation ; Humans ; Magnetic Field Therapy ; Melanoma ; enzymology ; pathology ; therapy ; Time Factors

To investigate whether lncRNA MALAT1 affects the migration and proliferation of breast cancer cells through the regulation with histone methyltransferase SMYD3, the endogenous MALAT1 in the MCF-7 and MDA-MB-231 breast cancer cells were knocked down by siRNA, and then the migration and proliferation of cells were detected by wound healing migration and MTT assay. The effects of si-MALAT1 on the mRNA and protein levels of miRNA-124, SMYD3 and its downstream genes were detected by Real time PCR and Western blot. The results showed that siRNA-targeted knockdown of MALAT1 reduced the migration and proliferation of breast cancer cells, and inhibited the transcriptional expression of SMYD3 and its downstream genes, including N-cadherin, MYL9, MMP9 and CYR61, and up-regulated miR-124. Overexpression of miR-124 reduced the expression of SMYD3 in breast cancer cells, and knockdown of MALAT1 attenuated the promotion of SMYD3 protein expression by miR-124 inhibitors. In addition, SMYD3 overexpression activated MALAT1 transcription, whereas siRNA interference with SMYD3 downregulated MALAT1. These results indicate that LncRNA MALAT1 acted as a competing endogenous RNA(ceRNA)of miR-124 to regulate expression of SMYD3 in breast cancer cells, and SMYD3 can activate the transcription of MALAT1, which will affect the proliferation and migration of breast cancer cells.