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 WS 582-2017 Specifications for testing of quality control in X and γ ray stereotactic radiotherapy system (hereinafter referred to as WS 582-2017) with respect to its current status, existing problems and evaluation since its implementation in Beijing, Chongqing, Jiangsu and Sichuan. Methods:The evaluation of this standard was carried out by means of field survey, questionnaire survey and expert seminar. In accordance with the WS 582-2017 implementation assessment programme, a total of 153 professionals involved in the implementation of WS 582-2017 from 80 radiological health technical service institutions, health supervision institutions and medical institutions of two municipalities and two provinces were surveyed in 2023. The investigation was carried out by using a combination of cluster sampling and stratified sampling and through sampling professionals with of different institution characteristics and different technical titles. The general survey method was used for Jiangsu province, while the cluster sampling method was used for others, with 153 individulas from 80 institutions being sampled for the investigation.Results:The awareness rate of WS 582-2017 was 80.39%, of which 90.85% considered it to be scientific and rigorous, 81.05% considered it to be operational and 97.39% considered it to be in consistency with the relevant regulatory standards, 90.20% considerd it to have played an important role in promoting radiological protection optimization for radiotherapy, 90.85% considered the implementation of it to have played a significant role in the quality control of standarded testing of radiotherapy equipment, and 86.93% considered the implementation of it to have played a significant role in enhancing the professional competence of relevant technicians.Conclusions:The overall implementation of WS 582-2017 in two municipalities and two provinces is good, with scientific and operable technical contents and the remarkable effect achieved in its implementation. However, the WS 582-2017 needs to be further improved in some areas, and training on the standard needs to be strengthened to improve the awareness rate of it.

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.

Objective:To investigate WS 582-2017 Specifications for testing of quality control in X and γ ray stereotactic radiotherapy system (hereinafter referred to as WS 582-2017) with respect to its current status, existing problems and evaluation since its implementation in Beijing, Chongqing, Jiangsu and Sichuan. Methods:The evaluation of this standard was carried out by means of field survey, questionnaire survey and expert seminar. In accordance with the WS 582-2017 implementation assessment programme, a total of 153 professionals involved in the implementation of WS 582-2017 from 80 radiological health technical service institutions, health supervision institutions and medical institutions of two municipalities and two provinces were surveyed in 2023. The investigation was carried out by using a combination of cluster sampling and stratified sampling and through sampling professionals with of different institution characteristics and different technical titles. The general survey method was used for Jiangsu province, while the cluster sampling method was used for others, with 153 individulas from 80 institutions being sampled for the investigation.Results:The awareness rate of WS 582-2017 was 80.39%, of which 90.85% considered it to be scientific and rigorous, 81.05% considered it to be operational and 97.39% considered it to be in consistency with the relevant regulatory standards, 90.20% considerd it to have played an important role in promoting radiological protection optimization for radiotherapy, 90.85% considered the implementation of it to have played a significant role in the quality control of standarded testing of radiotherapy equipment, and 86.93% considered the implementation of it to have played a significant role in enhancing the professional competence of relevant technicians.Conclusions:The overall implementation of WS 582-2017 in two municipalities and two provinces is good, with scientific and operable technical contents and the remarkable effect achieved in its implementation. However, the WS 582-2017 needs to be further improved in some areas, and training on the standard needs to be strengthened to improve the awareness rate of it.

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 investigate the influence of polycystic ovary syndrome (PCOS) patients with different body mass index (BMI) on perinatal and neonatal outcomes of frozen-thawed embryo transfer.Methods:A retrospective cohort study was performed on the clinical data of patients with PCOS infertility who underwent cryopreservation transplantation in Reproductive Center of the First Affiliated Hospital of Anhui Medical University from 2016 to 2020. The clinical pregnancy was singleton, a total of 1 481 cycles were divided into 4 groups according to BMI value. There were 75 cycles in the underweight group (BMI<18.5 kg/m 2), 793 cycles in the normal weight group (18.5 kg/m 2≤BMI<24.0 kg/m 2), 468 cycles in the overweight group (24.0 kg/m 2≤BMI<28.0 kg/m 2), 145 cycles in the obese group (BMI≥28.0 kg/m 2). The differences of general information, perinatal outcome and neonatal outcome were compared among the four groups. Results:Compared with the overweight group, the normal weight group and the underweight group, the obesity group had the highest early abortion rate [23.4% (34/145) vs. 15.8% (74/468) vs. 14.0% (111/793) vs. 9.3% (7/75), P=0.014], and the lowest live birth rate [68.3% (99/145) vs. 76.7% (359/468) vs. 79.7% (632/793) vs. 88.0% (66/75), P=0.003]. The incidence of gestational diabetes in the obesity group and the overweight group [6.9% (10/145) and 4.5% (21/468)] was higher than that in the normal weight group [2.3% (18/793)] (the obesity group vs. the normal weight group P=0.005, the overweight group vs. the normal weight group P=0.028). The rate of cesarean section in the obesity group and the overweight group [81.8% (81/99), 74.9% (269/359)] was higher than that in the normal weight group [67.6% (427/632)] and the underweight group [57.6% (38/66), the obesity group vs. the normal weight group P=0.005, the obesity group vs. the underweight group P=0.001, the overweight group vs. the normal weight group P=0.015, the overweight group vs. the underweight group P=0.004]. The macrosomia birth rate [18.2% (18/99), 15.6% (56/359)] was also higher than that of the normal weight group [10.1% (64/632)] and the underweight group [6.1% (4/66), the obesity group vs. the normal weight group P=0.018, the obesity group vs. the underweight group P=0.025, the overweight group vs. the normal weight group P=0.011, the overweight group vs. the underweight group P=0.041]. There were no significant differences in late abortion rate, gestational hypertension, ectopic pregnancy and premature birth rate, Apgar score, height and birth defects (all P>0.05). Conclusion:Obesity and overweight affect the perinatal outcomes and neonatal outcomes in patients with PCOS. In clinical work, attention should be paid to the weight management of PCOS patients.

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 investigate the influence of polycystic ovary syndrome (PCOS) patients with different body mass index (BMI) on perinatal and neonatal outcomes of frozen-thawed embryo transfer.Methods:A retrospective cohort study was performed on the clinical data of patients with PCOS infertility who underwent cryopreservation transplantation in Reproductive Center of the First Affiliated Hospital of Anhui Medical University from 2016 to 2020. The clinical pregnancy was singleton, a total of 1 481 cycles were divided into 4 groups according to BMI value. There were 75 cycles in the underweight group (BMI<18.5 kg/m 2), 793 cycles in the normal weight group (18.5 kg/m 2≤BMI<24.0 kg/m 2), 468 cycles in the overweight group (24.0 kg/m 2≤BMI<28.0 kg/m 2), 145 cycles in the obese group (BMI≥28.0 kg/m 2). The differences of general information, perinatal outcome and neonatal outcome were compared among the four groups. Results:Compared with the overweight group, the normal weight group and the underweight group, the obesity group had the highest early abortion rate [23.4% (34/145) vs. 15.8% (74/468) vs. 14.0% (111/793) vs. 9.3% (7/75), P=0.014], and the lowest live birth rate [68.3% (99/145) vs. 76.7% (359/468) vs. 79.7% (632/793) vs. 88.0% (66/75), P=0.003]. The incidence of gestational diabetes in the obesity group and the overweight group [6.9% (10/145) and 4.5% (21/468)] was higher than that in the normal weight group [2.3% (18/793)] (the obesity group vs. the normal weight group P=0.005, the overweight group vs. the normal weight group P=0.028). The rate of cesarean section in the obesity group and the overweight group [81.8% (81/99), 74.9% (269/359)] was higher than that in the normal weight group [67.6% (427/632)] and the underweight group [57.6% (38/66), the obesity group vs. the normal weight group P=0.005, the obesity group vs. the underweight group P=0.001, the overweight group vs. the normal weight group P=0.015, the overweight group vs. the underweight group P=0.004]. The macrosomia birth rate [18.2% (18/99), 15.6% (56/359)] was also higher than that of the normal weight group [10.1% (64/632)] and the underweight group [6.1% (4/66), the obesity group vs. the normal weight group P=0.018, the obesity group vs. the underweight group P=0.025, the overweight group vs. the normal weight group P=0.011, the overweight group vs. the underweight group P=0.041]. There were no significant differences in late abortion rate, gestational hypertension, ectopic pregnancy and premature birth rate, Apgar score, height and birth defects (all P>0.05). Conclusion:Obesity and overweight affect the perinatal outcomes and neonatal outcomes in patients with PCOS. In clinical work, attention should be paid to the weight management of PCOS patients.

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.