Neutralizing antibodies have been designed to specifically target and bind to the receptor binding domain (RBD) of spike (S) protein to block severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus from attaching to angiotensin converting enzyme 2 (ACE2). This study reports a distinctive nanobody, designated as VHH21, that directly catalyzes the S-trimer into an irreversible transition state through postfusion conformational changes. Derived from camels immunized with multiple antigens, a set of nanobodies with high affinity for the S1 protein displays abilities to neutralize pseudovirion infections with a broad resistance to variants of concern of SARS-CoV-2, including SARS-CoV and BatRaTG13. Importantly, a super-resolution screening and analysis platform based on visual fluorescence probes was designed and applied to monitor single proteins and protein subunits. A spontaneously occurring dimeric form of VHH21 was obtained to rapidly destroy the S-trimer. Structural analysis via cryogenic electron microscopy revealed that VHH21 targets specific conserved epitopes on the S protein, distinct from the ACE2 binding site on the RBD, which destabilizes the fusion process. This research highlights the potential of VHH21 as an abzyme-like nanobody (nanoabzyme) possessing broad-spectrum binding capabilities and highly effective anti-viral properties and offers a promising strategy for combating coronavirus outbreaks.
Single-Domain Antibodies/immunology* ; Spike Glycoprotein, Coronavirus/metabolism* ; SARS-CoV-2/immunology* ; Animals ; Humans ; Antibodies, Neutralizing/immunology* ; Camelus ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Angiotensin-Converting Enzyme 2

Objective:To investigate the effect of regional leptomeningeal collateral circulation (rLMC) score based on CT angiography (CTA) and onset-to-reperfusion time (OTR) on the outcome after endovascular treatment (EVT) in patients with anterior circulation acute large vessel occlusive stroke (ACA-LVOS).Methods:Patients with ACA-LVOS underwent EVT in the Department of Neurology, the Affiliated Hospital of Yangzhou University from July 2017 to July 2023 were included retrospectively. The rLMC score 0-10 was defined as poor collateral circulation, and 11-20 were defined as good collateral circulation. At 90 days after EVT, the modified Rankin Scale (mRS) was used to evaluate the outcome. A score of 0-2 was defined as a good outcome and 3-6 were defined as a poor outcome. Multivariate logistic regression analysis was used to determine the independent influencing factors of the outcome after EVT. Results:A total of 144 patients with ACA-LVOS underwent EVT were enrolled, including 78 males (54.2%), median aged 73 years. The median baseline National Institutes of Health Stroke Scale (NIHSS) score was 16, the median baseline Alberta Stroke Program Early CT Score (ASPECTS) was 9, and the median OTR was 330.5 minutes. Eighty patients (55.6%) had good collateral circulation, 63 (43.8%) had poor outcome, including 13 deaths. Univariate analysis showed that there were significant differences in hypertension, previous stroke history, smoking, triglycerides, baseline NIHSS score, baseline ASPECTS, OTR, and collateral circulation status between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that good collateral circulation (odds ratio [ OR] 0.223, 95% confidence interval [ CI] 0.077-0.643; P=0.005) was an independent predictor of good outcome. In the poor collateral circulation group, longer OTR was an independent predictor of poor outcome ( OR 1.020, 95% CI 1.008-1.032; P=0.001). In the good collateral circulation group, longer OTR was not an independent risk factor for poor outcome ( OR 1.005, 95% CI 1.000-1.010; P=0.062). Conclusion:rLMC score based on CTA and OTR are the independent predictors of the outcome after EVT in patients with ACA-LVOS.

Objective:To explore effect of polydatin on ocular surface dysfunction and pathology in dry eye disease(DED)rats and its protective mechanism.Methods:Eighteen from 90 SD rats were randomly chosen as control group,and other rats were injected subcutaneously with scopolamine hydrobromide to establish DED model.After modeling,rats were separated into model group,0.05%polydatin group,0.5%polydatin group,0.5%polydatin+U0126[mitogen-activated protein kinase kinase(MEK)inhibi-tor]group,with 18 rats in each group;each administration group was given corresponding doses of drugs for intervention,and rats in control group and model group were given same amount of normal saline.On the 7th,14th,21st,and 28th day of intervention,Schirmer test was performed to measure tear secretion amount of rats in each group,fluorescein staining was performed to measure tear film breakup time(BUT)and corneal damage of rats in each group;RT-qPCR was performed to measure mRNA expression of pro-inflammatory factors TNF-α,IFN-γ and IL-1β in cornea and conjunctival tissues;HE staining was performed to observe histopatho-logical changes of cornea;PAS staining was performed to evaluate number of conjunctival goblet cells;Western blot was performed to detect expressions of MEK/extracellular regulatory protein kinase(ERK)pathway related proteins.Results:Compared with control group,tear secretion amount BUT and number of conjunctival goblet cells in model group were obviously reduced(P<0.05),corneal injury score,TNF-α,IFN-γ and IL-1β mRNA levels were obviously increased(P<0.05),corneal epithelial layer was thinned,a large number of inflammatory cell infiltration and new capillaries could be seen;compared with model group,tear secretion amount,BUT,number of conjunctival goblet cells,p-MEK1/2/MEK1/2 and p-ERK1/2/ERK1/2 in 0.05%polydatin group and 0.5%polydatin group were obviously increased(P<0.05),corneal injury score,TNF-α,IFN-γ and IL-1β mRNA levels were obviously reduced(P<0.05),pathological damage of corneal tissue was obviously relieved;U0126 could significantly attenuate protective effects of polydatin on ocular surface dysfunction and pathological changes in DED rats.Conclusion:Polydatin may improve ocular surface dysfunction and pathology of DED rats induced by scopolamine by activating MEK/ERK pathway.

BACKGROUND: Cholecystectomy is a standard surgery for patients suffering from gallbladder diseases, while the causal effects of cholecystectomy on colorectal cancer (CRC) and other complications are still unknown. METHODS: We obtained genetic variants associated with cholecystectomy at a genome-wide significant level ( P value <5 × 10 -8 ) as instrumental variables (IVs) and performed Mendelian randomization (MR) to identify the complications of cholecystectomy. Furthermore, the cholelithiasis was also treated as the exposure to compare its causal effects to those of cholecystectomy, and multivariable MR analysis was carried out to judge whether the effect of cholecystectomy was independent of cholelithiasis. The study was reported based on Strengthening the Reporting of Observational Studies in Epidemiology Using Mendelian Randomization guidelines. RESULTS: The selected IVs explained 1.76% variance of cholecystectomy. Our MR analysis suggested that cholecystectomy cannot elevate the risk of CRC (odds ratio [OR] =1.543, 95% confidence interval [CI]: 0.607-3.924). Also, it was not significant in either colon or rectum cancer. Intriguingly, cholecystectomy might decrease the risk of Crohn's disease (OR = 0.078, 95% CI: 0.016-0.368) and coronary heart disease (OR = 0.352, 95% CI: 0.164-0.756). However, it might increase the risk of irritable bowel syndrome (IBS) (OR = 7.573, 95% CI: 1.096-52.318). Cholelithiasis could increase the risk of CRC in the largest population (OR = 1.041, 95% CI: 1.010-1.073). The multivariable MR analysis suggested that genetic liability to cholelithiasis could increase the risk of CRC in the largest population (OR = 1.061, 95% CI: 1.002-1.125) after adjustment of cholecystectomy. CONCLUSIONS The study indicated that cholecystectomy might not increase the risk of CRC, but such a conclusion needs further proving by clinical equivalence. Additionally, it might increase the risk of IBS, which should be paid attention to in clinical practice.
Humans ; Mendelian Randomization Analysis ; Irritable Bowel Syndrome ; Colorectal Neoplasms/genetics* ; Cholelithiasis/complications* ; Cholecystectomy/adverse effects* ; Genome-Wide Association Study ; Polymorphism, Single Nucleotide

Objective:To reduce the immunogenicity of vaccinia virus vector by replacing the D8L region, which is a neutralizing antibody epitope in vaccinia virus, with an exogenous gene.Methods:A gene fragment encoding influenza virus hemagglutinin (HA) was inserted into the D8L region to replace it using homologous recombination technique. Then, a recombinant vaccinia virus influenza vaccine was constricted. A recombinant vaccinia virus vaccine with the TK region expressing HA was used as a control. The expression of HA was validated by Western blot. BALB/c mice were immunized with the vaccines and the serum antibody titers two weeks after each immunization were evaluated by ELISA and hemagglutination inhibition assay. The protective efficacy of the recombinant vaccinia virus was assessed through a challenge experiment.Results:Western blot confirmed the successful expression of HAD8L protein in the constructed recombinant vaccines. ELISA and hemagglutination inhibition assay showed that after the primary immunization, the anti-HA antibody titer induced by the recombinant vaccinia virus with D8L region mutation was slightly higher than that induced by the vaccine with TK region mutation, and the difference was statistically significant with the increase of immunization times ( P<0.05). The recombinant vaccinia virus with D8L region mutation showed significantly lower immunogenicity than the recombinant virus with TK region mutation after the primary immunization, but there was no significant difference between them with the increase of immunization times ( P>0.05). After H1N1pdm challenge, no virus was detected in the mice immunized with the recombinant vaccinia virus with D8L region mutation and the mice showed mild lung inflammation and less tissue damage. Conclusions:This study indicated that inserting exogenous genes into the D8L region of the neutralizing antibody epitope in the vaccinia virus vector could help to reduce the immunogenicity of the vector itself and enhance the immunogenicity of the exogenous genes. This provided a reference for the use of the vaccinia virus vector as a delivery tool in the field of vaccines or gene therapy.

Objective:To explore the effect of closed-loop nursing on self-efficacy and nursing satisfaction in children with nephrotic syndrome (NS) .Methods:From March 2019 to January 2021, convenience sampling was used to select 104 children with NS and their accompanying parents in Henan Provincial People's Hospital as the research object. The children were divided into the control group and the observation group by random number table method, 52 cases in each group. The control group received routine nursing, and the observation group carried out closed-loop nursing on the basis of the control group. The scores of the General Self-efficacy Scale and Satisfaction Scale were compared between the two groups.Results:After three months of intervention, the children's self-efficacy score in the observation group was higher than that in the control group, and the parents' satisfaction with nursing was 94.23% (49/52) , which was higher than 78.85% (41/52) in the control group. The differences were all statistically significant ( P<0.05) . Conclusions:The closed-loop nursing can improve the self-efficacy of children with NS and the satisfaction of their parents, which is worthy of clinical promotion.

Background: Studies have shown that curcumin can regulate the expressions of a variety of miRNAs and affect tumor cell biological behavior. However, whether curcumin affects the biological behavior of gastric cancer cells by regulating expression of miR-539 has not been clarified. Aims: To investigate the effect and mechanism of curcumin on proliferation, migration, invasion, apoptosis of gastric cancer cells by regulating the expression of miR-539. Methods: A total of 49 gastric cancer tissue specimens were collected. Gastric cancer AGS, SGC7901 cells were divided into blank group, curcumin group, negative control group, miR-539 mimics group, miR-539 inhibitor group, inhibitor control group, miR-539 inhibitor +curcumin group. qRT-PCR was used to detect the mRNA expression of miR-539 in gastric cancer tissue and cells. Cell proliferation was detected by MTT. Scratch test, Transwell test were used to detected cell migration and invasion. Cell apoptosis was determined by flow cytometry. Western blotting was used to detect protein expressions of APOBEC3B, c-Myc, cyclin D1, claudin-1 and N-cadherin. Results: Expressions of miR-539 mRNA in gastric cancer tissue and gastric cancer cells were significantly decreased than those in corresponding controls (P<0.05), and was related to tumor stage in gastric cancer patients (P<0.05). Curcumin up-regulated the expression of miR-539 mRNA in a dose-dependent manner (P<0.05). Compared with corresponding control group, cell proliferation, migration and invasion were significantly decreased in miR-539 mimics group and curcumin group (P<0.05), cell apoptosis rate was significantly increased (P<0.05), and protein expressions of APOBEC3B, c-Myc, cyclin D1, claudin-1 and N-cadherin were significantly decreased (P<0.05). Conclusions: Expression of miR-539 is decreased in gastric cancer, and curcumin can inhibit proliferation, migration and invasion of gastric cancer cells and induce cell apoptosis by up-regulating the expression of miR-539.

Objective:To investigate the value of D-dimer, carbohydrate antigen 199 (CA199) and insulin-like growth factor binding protein 2 (IGFBP2) for postoperative monitoring and prediction of survival time in patients with resectable pancreatic cancer.Methods:The data of 119 patients with pancreatic cancer who were admitted to the First Hospital of Qinhuangdao from May 2010 to January 2014 were collected. Immunoturbidimetry was used to determine the level of D-dimer before surgery, at postoperative stable disease stage and disease progression stage; electrochemiluminescence was used to determine the level of CA199, and enzyme-linked immunosorbent assay (ELISA) was used to determine the serum IGFBP2 level. A total of 30 healthy people and 40 patients with pancreatic serous cystadenoma were treated as the controls. The correlations of the levels of preoperative D-dimer, CA199 and IGFBP2 with clinicopathological characteristics and survival time of patients with pancreatic cancer were analyzed.Results:The levels of preoperative D-dimer, CA199, IGFBP2 in patients with pancreatic cancer were higher than those in the both control group (all P < 0.01). The levels of serum D-dimer, CA199 and IGFBP2 after the progression of pancreatic cancer were higher than those at postoperative stable disease stage [1 496.0 ng/ml (590.0 ng/ml, 2 280.4 ng/ml) vs. 578.1 ng/ml (381.7 ng/ml, 671.5 ng/ml), 207.0 U/ml (54.5 U/ml, 736.5 U/ml) vs. 31.9 U/ml (14.1 U/ml, 44.0 U/ml), (435±107) ng/ml vs. (249±83) ng/ml, all P < 0.01]. There were no statistical differences in the proportion of pancreatic cancer patients stratified by different clinicopathological factors with the increased levels of D-dimer before operation (all P > 0.05). The proportion of the increased levels of CA199 and IGFBP2 in patients with lymph node metastasis was higher than that in patients without lymph node metastasis (both P < 0.05); there was no association of the increased levels of CA199 and IGFBP2 with other factors (all P > 0.05). The preoperative progression-free survival (PFS) time and overall survival (OS) time of pancreatic cancer patients with elevated D-dimer level was shorter than that for those with normal D-dimer level [(10.6±1.2) months vs. (20.4±2.4) months, (18.9±1.9) months vs. (29.2±2.6) months, both P < 0.01]. When the threshold value of CA199 was 37 U/ml, there was no correlation between CA199 and survival of pancreatic cancer patients (all P > 0.05); when the threshold value was 253.8 U/ml (median CA199 for the enrolled patients) and 1 000 U/ml, patients with elevated CA199 level had shorter OS time and PFS time compared with the patients with normal CA199 level [253.8 U/ml: (11.5±1.5) months vs. (21.0±2.6) months, (19.9±2.1) months vs. (29.0±2.7) months, both P < 0.01; 1 000 U/ml: (8.9±1.9) months vs. (19.1±1.9) months, (15.5±2.3) months vs. (28.0±2.0) months, both P < 0.01]. When the threshold value of IGFBP2 was 339.1 ng/ml, patients with elevated preoperative IGFBP2 level had shorter PFS time and OS time compared with the patients with normal IGFBP2 level [(10.8± 1.1) months vs. (21.1±2.6) months, (18.9±1.8) months vs. (30.3±2.8) months, both P < 0.01]. Cox multivariate analysis showed that preoperative D-dimer and IGFBP2 levels were independent factors affecting PFS and OS in patients with pancreatic cancer (D-dimer: HR = 0.561, 95% CI 0.336-0.936, P = 0.027; HR = 0.515, 95% CI 0.303-0.874, P = 0.014; IGFBP2: HR = 0.430, 95% CI 0.253-0.731, P = 0.002; HR = 0.361, 95% CI 0.202-0.644, P = 0.001). Conclusions:For patients with resectable pancreatic cancer, D-dimer, CA199 and IGFBP2 can be used for postoperative condition monitoring, and preoperative D-dimer and IGFBP2 can be used for survival time prediction.

Objective To explore the efficacy and safety of apatinib combined with S-1 in patients with advanced NSCLC without sensitive gene mutation or unknown mutation status.Methods One hundred and four patients with advanced NSCLC without sensitive gene mutation or unknown mutation status were selected from the oncology department of the First Hospital of Qinhuangdao City,Hebei Province from April 2015 to April 2017.All patients refused intravenous chemotherapy.One hundred and four patients were randomly divided into treatment group (apatinib combined with S-1 group) and control group (S-1 alone group) by 1:1 digital method.However,two patients in the treatment group transferred to the control group for personal reasons.There is 50 cases in apatinib combined with S-1 group and 54 cases in S-1 group.The efficacy and adverse reactions of the two groups were evaluated.Results The objective remission rate was 48.0％ (24/50) and 27.8％ (15/54) (x2=4.530,P =0.033),the disease control rate was 82.0％ (41/50) and 74.1％ (40/54) (x2=0.947,P=0.331),the median PFS was 6.6 months and 3.4 months (t=25.555,P =0.000),the median OS was 16.0 months and 10.5 months (t =59.439,P =0.000),respectively.The overall incidence of adverse reactions was 82.0％ (41/50) and 70.4％ (38/54) respectively (x2 =1.923,P=0.166),of which 18.0％ (9/50) and 13.0％ (7/54) were more than grade 3 respectively (x2 =0.506,P =0.477).There was no death caused by treatment-related adverse reactions in both groups.Conclusion Appatinib combined with S-1 capsule has good short-term and long-term efficacy in the treatment of advanced non-small cell lung cancer without gene mutation or unknown mutation.The adverse reactions are tolerable and can be used as first-line treatment for patients unwilling to receive intravenous chemotherapy.

Objective to explore the value of scenario simulation combined with PDCA cycle in the training of newly recruited staff of the radiology department. Methods thirty newly recruited members of the radiology department were selected as research subjects and received three months of training using scenario simulation combined with PDCA cycle teaching. The subjects took examinations before and after the training to estimate its effect on the their first-aid related knowledge and skills, as well as the apprehension of professional knowledge; questionnaires were used to collect the subjects' self-evaluation and their feedbacks for the training. T test was conducted using SPSS 20.0. Results the scores of first aid related knowledge, first aid instrument use, first aid skills and first aid ability after training were significantly higher, and the differences were statistically significant. After the training, the staff's performance in terms of noun explanation, problem solving and essay questions was significantly higher than that before the training, and the overall performance was significantly improved. More than 90％of the new recruits believed that scenario simulation combined with PDCA had practical effects on improving clinical practice ability , knowledge understanding, comprehensive analysis, problem solving ability and cooperative ability. More than 85％ of the subjects considered it helpful for mastering the teaching content , improving learning efficiency , developing independent learning ability and stimulating learning interest. Conclusion scenario simulation teaching combined with PDCA can improve new employee's learning interest and teaching satisfaction, and further improve doctor-patient communication ability and training effect.