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 explore the cognitive characteristics, influencing factors, and treatment needs regarding biologic agents among patients with psoriasis.Methods:A cross-sectional study was conducted. Patients with psoriasis attending the Department of Dermatology, Xijing Hospital, Air Force Medical University were selected from October to December 2022, and from June to December 2023. A self-designed electronic questionnaire was used for investigation, covering demographic characteristics, psoriasis history (disease types, disease duration, previous treatments, etc.), biologics knowledge (sources of awareness, core cognitive dimensions), and treatment needs.Results:The valid questionnaire response rate reached 93.2% (439/471). The ages of enrolled patients were 35.95 ± 12.57 years, and the disease duration was 7.90 ± 3.26 years. Psoriasis vulgaris was the predominant type (363 cases, 82.69%). The overall awareness rate of biologics slightly increased from 68.62% (105/153) in 2022 to 72.38% (207/286) in 2023 ( P > 0.05). Primary information sources included new media (WeChat/internet) platforms (168 cases, 53.84%) and peer-to-peer sharing (115 cases, 36.86%), while physician counseling merely accounted for 9.29% (29 cases) ( P < 0.001). Insufficient knowledge of biologics was manifested primarily as poor awareness of comorbidities (47.60%, 209/439) and treatment monitoring protocols (22.32%, 98/439). Core concerns regarding biologic therapy included safety (73.34%, 322/439), economic burden (65.14%, 286/439), and long-term efficacy (63.55%, 279/439) ; 60.13% (264/439) of the patients expected rapid improvement of skin symptoms. As for treatment modalities, 90.20% (396/439) of the patients preferred regimens with extended dosing intervals. Conclusions:The patients with psoriasis demonstrated an imbalance in their cognitive structure regarding biologic agents. Their treatment needs exhibited multidimensional characteristics, emphasizing not only rapid clearance of skin lesions but also greater importance of treatment safety and cost-effectiveness.

OBJECTIVE To investigate the horizontal transfer mechanisms and genetic evolutionary characteristics of virulence genes in hypervirulent Klebsiella pneumoniae(HvKP)and resistance genes in carbapenem-resistant K.pneumoniae(CRKP).METHODS The donor strain was a CRKP strain harboring both VIM and KPC resist-ance genes.The positive control was a CRKP strain containing blaIMP-4.The recipient strain was Escherichia coli J53.The horizontal transfer capability of the resistance genes was verified by plasmid conjugation assay.Fifty HvKP strains were collected from the First Affiliated Hospital of Jinan University from 2017 to 2021 and conjuga-ted with the plasmids of five CRKP strains which could mediate the transmission of drug resistance genes.The conjugants were screened by selective medium containing meropenem,and polymerase chain reaction(PCR)was used to detect the resistance and virulence genes in the conjugation-successful strains.The lethality of conjugants was verified by Galleria mellonella larval assays.RESULTS The donor CRKP strain had the ability to mediate re-sistance gene transmission.Its blaKPC gene was successfully transferred to E.coli J53,but the transference of the VIM gene failed.Five CRKP strains with the ability to mediate resistance gene transmission were screened,of which one strain carried both VIM and KPC resistance genes,and the other four strains carried the IMP-4 resist-ance gene.Only one CRKP strain with the blaIMP-4 resistance gene was successfully conjugated with HvKP,carry-ing both virulence genes(aerobactin and RmpA)and the blaIMP-4 resistance gene,with positive wire drawing test result.The Galleria mellonella larvae test showed that the conjugation-successful strain caused the death of all lar-vae within 24 hours.CONCLUSIONS In the hospital environment,CRKP and HvKP can form"superbugs"with both drug resistance and high virulence through plasmid conjugation,posing a severe challenge to clinical an-ti-infection treatment.It is necessary to strengthen the prevention and control of hospital-acquired infections.

Objective:To explore the cognitive characteristics, influencing factors, and treatment needs regarding biologic agents among patients with psoriasis.Methods:A cross-sectional study was conducted. Patients with psoriasis attending the Department of Dermatology, Xijing Hospital, Air Force Medical University were selected from October to December 2022, and from June to December 2023. A self-designed electronic questionnaire was used for investigation, covering demographic characteristics, psoriasis history (disease types, disease duration, previous treatments, etc.), biologics knowledge (sources of awareness, core cognitive dimensions), and treatment needs.Results:The valid questionnaire response rate reached 93.2% (439/471). The ages of enrolled patients were 35.95 ± 12.57 years, and the disease duration was 7.90 ± 3.26 years. Psoriasis vulgaris was the predominant type (363 cases, 82.69%). The overall awareness rate of biologics slightly increased from 68.62% (105/153) in 2022 to 72.38% (207/286) in 2023 ( P > 0.05). Primary information sources included new media (WeChat/internet) platforms (168 cases, 53.84%) and peer-to-peer sharing (115 cases, 36.86%), while physician counseling merely accounted for 9.29% (29 cases) ( P < 0.001). Insufficient knowledge of biologics was manifested primarily as poor awareness of comorbidities (47.60%, 209/439) and treatment monitoring protocols (22.32%, 98/439). Core concerns regarding biologic therapy included safety (73.34%, 322/439), economic burden (65.14%, 286/439), and long-term efficacy (63.55%, 279/439) ; 60.13% (264/439) of the patients expected rapid improvement of skin symptoms. As for treatment modalities, 90.20% (396/439) of the patients preferred regimens with extended dosing intervals. Conclusions:The patients with psoriasis demonstrated an imbalance in their cognitive structure regarding biologic agents. Their treatment needs exhibited multidimensional characteristics, emphasizing not only rapid clearance of skin lesions but also greater importance of treatment safety and cost-effectiveness.

OBJECTIVE To investigate the horizontal transfer mechanisms and genetic evolutionary characteristics of virulence genes in hypervirulent Klebsiella pneumoniae(HvKP)and resistance genes in carbapenem-resistant K.pneumoniae(CRKP).METHODS The donor strain was a CRKP strain harboring both VIM and KPC resist-ance genes.The positive control was a CRKP strain containing blaIMP-4.The recipient strain was Escherichia coli J53.The horizontal transfer capability of the resistance genes was verified by plasmid conjugation assay.Fifty HvKP strains were collected from the First Affiliated Hospital of Jinan University from 2017 to 2021 and conjuga-ted with the plasmids of five CRKP strains which could mediate the transmission of drug resistance genes.The conjugants were screened by selective medium containing meropenem,and polymerase chain reaction(PCR)was used to detect the resistance and virulence genes in the conjugation-successful strains.The lethality of conjugants was verified by Galleria mellonella larval assays.RESULTS The donor CRKP strain had the ability to mediate re-sistance gene transmission.Its blaKPC gene was successfully transferred to E.coli J53,but the transference of the VIM gene failed.Five CRKP strains with the ability to mediate resistance gene transmission were screened,of which one strain carried both VIM and KPC resistance genes,and the other four strains carried the IMP-4 resist-ance gene.Only one CRKP strain with the blaIMP-4 resistance gene was successfully conjugated with HvKP,carry-ing both virulence genes(aerobactin and RmpA)and the blaIMP-4 resistance gene,with positive wire drawing test result.The Galleria mellonella larvae test showed that the conjugation-successful strain caused the death of all lar-vae within 24 hours.CONCLUSIONS In the hospital environment,CRKP and HvKP can form"superbugs"with both drug resistance and high virulence through plasmid conjugation,posing a severe challenge to clinical an-ti-infection treatment.It is necessary to strengthen the prevention and control of hospital-acquired infections.

Neck dissection and reconstruction are two important aspects of oral cancer treatment.There are various surgical methods for neck dissection and reconstruction,but all of them are performed by open surgery.This article reports a full endoscopic neck dis-section through the retroauricular hairline approach,the radical resection of the intraoral tumor and the repair of the defect by superfi-cial circumflex iliac artery perforator flap with in situ vascular anastomosis intraorally.The incision is located in the hairline,hidden and invisible,and there is no exposed surgical scar on the neck after surgery.This paper introduces the technique of scarless neck dissection combined with free skin flap repair for the treatment of oral cancer and discusses its advantages and disadvantages.

Objective To evaluate iron deposition in gray matter nuclei in patients with cerebral small vessel disease(CSVD)based on quantitative susceptibility mapping(QSM)and to analyze its correlation with cognitive function.Methods Patients with CSVD attending the outpatient clinic in the Department of Neurology at Xuanwu Hospital of Capital Medical University from December 2016 to November 2022,and healthy controls recruited from previous studies in the Department of Radiology and Nuclear Medicine at Xuanwu Hospital of Capital Medical University from September 2022 to November 2022 were retrospectively consecutively collected.Baseline data of CSVD patients and healthy controls was collected and compared,including age,sex,past history(hypertension,diabetes,hyperlipidemia),smoking history,alcohol consumption history and Montreal cognitive assessment(MoCA)scale score.MRI of all CSVD patients and healthy controls were collected,including three-dimensional T1 weighted imaging,QSM,T2 weighted imaging,and fluid attenuated inversion recovery(FLAIR)sequence imaging.According to the MRI-related imaging features and CSVD total burden score,the patients were divided into mild CSVD(CSVD-m)group and severe CSVD(CSVD-s)group,and healthy controls were the control group.QSM was used to obtain the susceptibility values of gray matter nuclei for all CSVD patients and controls.One-way covariance analysis and Bonferroni correction were used to compare the gray matter nuclei susceptibility values among the three groups.Spearman correlation analysis was performed between susceptibility values of gray matter nuclei with statistically significant differences in susceptibility values and cognitive function.Results A total of 61 cases of CSVD patients were included,including 29 cases in the CSVD-s group and 32 cases in the CSVD-m group;32 healthy controls were included in the control group.(1)There was no statistically significant difference in age,sex,hypertension,diabetes,hyperlipidemia,smoking,and alcohol consumption between the CSVD-s group,CSVD-m group and control group(all P＞0.05).The MoCA scale scores of the CSVD-s group and CSVD-m group were lower than those of the control group(25.0[22.5,27.5]points,27.0[25.0,29.0]points than 28.0[27.0,29.0]points,H=15.006,P＜0.01).The difference in the imaging features distribution of cerebral microbleeds,white matter hyperintensity,and perivascular space among the CSVD-s group and the CSVD-m group was statistically significant(all P＜0.05).(2)The differences in susceptibility values of the left putamen(F=4.790),pallidus(F=12.896),hippocampus(F=3.904)and the right putamen(F=36.278),pallidus(F=39.449),caudate nucleus(F=6.797),and thalamus(F=6.525)were statistically significant among the three groups(all P＜0.05).After Bonferroni correction,the susceptibility values of the left putamen and pallidus and the right putamen,pallidus,caudate nucleus,and thalamus in the CSVD-s group were higher than those of the control group(all P＜0.05);the susceptibility values of the left pallidus and the right pallidus,putamen,and thalamus in the CSVD-m group were higher than those of the control group(all P＜0.01),and the left hippocampus was lower than that of the control group(P=0.045).(3)The susceptibility values of the bilateral putamen were significantly negatively correlated with MoCA scale score(left putamen:rs=-0.316,P=0.015;right putamen:rs=-0.316,P=0.014).Conclusion Abnormal iron metabolism occurs in gray matter nuclei of CSVD patients,and iron deposition in the putamen correlate with cognitive dysfunction.

Objective To construct a non-drug intervention program for acute chemotherapy-related nausea and vomiting in children with cancer and to evaluate its efficacy.Methods Through literature review and Delphi expert correspondence,a non-drug intervention program for acute chemotherapy-related nausea and vomiting in children with cancer was constructed.By the convenience sampling method,200 consecutive children who received chemotherapy in the neurosurgery department of a tertiary children's hospital in Zhejiang province from February 1 to October 31,2023 were included as the application subjects,with 100 cases in an experimental group and 100 cases in a control group.The experimental group applied the non-drug intervention program of acute chemotherapy-related nausea and vomiting in children with cancer,and the routine measures were applied in the control group.The incidence of nausea and vomiting,severity of vomiting,compliance rate of normal sleep duration and incidence of negative emotions were compared between the 2 groups.Results The recovery rate of the valid questionnaire in 2 rounds of expert letter inquiry was 100％,and the expert authority coefficient was 0.836.The Kendall harmony coefficients were 0.471 and 0.820(P＜0.001),and the final non-drug intervention program for pediatric acute chemotherapy-related nausea and vomiting included 5 primary,14 secondary and 18 tertiary items.The results showed that the incidence of nausea,vomiting and negative emotions in the experimental group were lower than that in the control group,with statistically significant differences(P＜0.05).The severity of vomiting was less than it in the control group,with statistically significant difference(P＜0.05).The standard rate of normal sleep time was higher than that of the control group,and the difference was statistically significant(all P＜0.05).Conclusion The non-drug intervention program of chemotherapy-related nausea and vomiting in children is scientific and feasible,and the implementation of the program can reduce the incidence of nausea,vomiting and negative emotions,reduce the severity of vomiting,and improve the standard rate of normal bedtime in children.

In order to investigate the changes in lipid metabolism of the host after infection with porcine epidemic diarrhea virus(PEDV),blood,intestinal and cellular samples were collected from PEDV-infected piglets and Vero cells,and the levels of triglyceride(TG)and total cholesterol(TC)were detected by blood biochemical analyses,flow cytometry,and immunofluorescence,and changes in the gene levels of the enzymes related to lipid metabolism of the hosts were detected by qPCR.The results showed that PEDV infection significantly elevated host total lipid levels and sig-nificantly promoted the expression of enzymes related to fatty acid and cholesterol metabolism in the intestine and Vero cells of piglets.In conclusion,PEDV infection leads to an increase in host to-tal lipid levels,remodeling of lipid metabolism in piglets intestine and Vero cells,and lipid metabo-lism gene changes.The results of this study show that lipid metabolism-related genes can be used as a key target to influence PEDV replication,which will provide a new theoretical basis for the subsequent prevention,control and treatment of PEDV infection.

Objective To explore the effects of Manchester pain management on postoperative pain and recovery in pregnant women with gestational hypertension.Methods In total,134 pregnant women with gestational hypertension admitted to the Obstetrics Department of Shengjing Hospital between April 2022 and May 2023 were selected.The patients were randomly divided into a control group(group C)and a Manchester group(MPMM group).Pain frequency,pain(VAS),anxiety(SAS),depression(SDS),and sleep quality(PSQI)scores,as well as morning systolic blood pressure(SP),diastolic blood pressure(DP),mean arterial pressure(MAP),first time out of bed,and hospitalization time were recorded and compared between the two groups on days 0,1,and 3 after surgery.Results The pain score and frequency of pain in the MPMM group were lower than those in the C group on days 0 and 1 after surgery(P<0.05);the SAS and PSQI scores were lower in the C group on days 1 and 3 after surgery(P<0.05);and the SDS scores on days 0,1,and 3 after surgery were lower than those in the C group(P<0.05).On days 0,1,and 3 after surgery,the SP in the MPMM group was lower than that in the C group;the DP and MAP were lower than those in the C group on days 1 and 3 after surgery,and the first time of getting out of bed and hospitalization time were significantly shorter than those in group C(P<0.05).Conclusion The MPMM alleviates postoperative pain,reduces anxiety and depression,improves sleep quality,and promotes early recovery in pregnant women with gestational hypertension.