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 analyze the current status of clinical trials of Chinese materia medica for the purpose of registration in China; To provide reference for the research and development of new TCM drugs.Methods:Clinical trials of Chinese materia medica/natural medicine registered in Drug Clinical Trial Registration and Information Disclosure Platform were retrieved from inception to December 31, 2024. Excel 2019 software was used to input and analyze the data such as the number of registered clinical trials, date of first publication, study status, field of indication, trial phases, sponsors, group leader, and design types.Results:A total of 1 137 Chinese materia medica clinical trials had been registered, accounting for 4.12% of the total number registered on the platform. Phase Ⅱ clinical trials accounted for the highest proportion (58.8%), and 99.7% of clinical trials conducted domestically. The sponsors were predominantly domestically pharmaceutical enterprises. These 1 137 clinical trials of Chinese materia medica clinical trials involved 752 drug categories, 28 dosage forms, and 796 varieties (the same class of drugs had different drug dosage forms), with capsules being the most common. The indications primarily focused on respiratory, digestive, cardio-cerebrovascular, neuropsychiatric, gynecological diseases. The group leader of clinical trials was distributed in 28 provinces, among which the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine as the group leader, undertook the most clinical trials of TCM. 89.9% of the clinical trials adopted the randomized controlled trial design, and only 31.9% of the clinical trials purchased insurance for the subjects.Conclusion:The research and development of new TCM drugs has entered a phase of vigorous development. Further efforts are still needed in establishing systematic guidelines for Chinese materia medica clinical trials, accelerating the internationalization of TCM, exploring innovative dosage forms and indications, and strengthening the protection of participants' rights.

ObjectiveTo explore the therapeutic effect of Huangliansan on atopic dermatitis （AD） model mice induced by 2， 4-dinitrochlorobenzene （DNCB）. MethodA total of 42 male BALB/c mice were randomly divided into normal group， model group， hydrocortisone group， low， medium， and high-dose groups （0.3， 0.6， 1.2 g·kg^-1） of Huangliansan oil， and water extract group （0.6 g·kg^-1） of Huangliansan. In addition to the normal group， DNCB was applied on the back of mice in other groups to establish the AD model. On the 15^th day after DNCB stimulation， each group was given the corresponding drug or solvent， and the changes in skin lesions， dermatitis score， and frequency of scratching were observed and recorded. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the skin and spleen. Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA levels of filaggrin （FLG）， lorophane （LOR）， and involucrin （IVL） in skin， as well as immunoglobulin E （lgE）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and interferon-γ （IFN-γ） in spleen. ResultCompared with the normal group， the model group showed symptoms of skin swelling and scab， and the score of dermatitis， the frequency of scratching， and the spleen index were increased （P<0.05）. The expression levels of FLG， LOR， and IVL in skin tissue were significantly decreased （P<0.01）， and the mRNA expressions of IgE， IL-4， IL-6， IL-1β， and TNF-α in the spleen were significantly increased， while the expression level of IFN-γ was significantly decreased （P<0.01）. Compared with the model group， the symptoms of skin erythema， scaly， and scab of mice in each drug group were alleviated to varying degrees， and the score of dermatitis， the frequency of scratching， and the spleen index were decreased （P<0.05， P<0.01）. In addition， the expression levels of FLG， LOR， and IVL in the skin of mice in the drug group were increased （P<0.05， P<0.01）， and the mRNA expression of IgE， IL-4， IL-6， IL-1β， and TNF-α in spleen were decreased. IFN-γ was increased （P<0.05， P<0.01）， and the lesions of the skin and spleen were improved to varying degrees. The medium-dose group of Huangliansan oil and hydrocortisone group had the most obvious manifestations （P<0.05， P<0.01）. The indexes in the medium-dose group of Huangliansan oil were better than those in the water extract group of Huangliansan. ConclusionHuangliansan may improve the expression level of skin barrier protein， inhibit the expression of helper T cell 2 （Th2）-related inflammatory factors， increase the expression of helper T cell 1 （Th1） inflammatory factors， restore the skin barrier function and Th1/Th2 balance in the spleen， regulate the inflammatory response in the spleen of AD mice， and thus relieve AD. Huangliansan oil is more effective than water extract.

Objective:To investigate the efficacy and safety of dual wavelength pulsed dye laser combined with 30% supramolecular salicylic acid in the treatment of moderate and severe facial acne.Methods:Sixty patients with moderate and severe acne that visited the Dermatology Department of the First Affiliated Hospital of Chongqing Medical University from May 2020 to January 2021, were selected and randomly divided into observation group and control group, with 30 patients in each group. The observation group was given dual-wavelength pulsed dye laser combined with 30% supramolecular salicylic acid. 30% supramolecular salicylic acid was used once every two weeks, for a total of six times. Dual-wavelength pulsed dye laser was given once a month, for a total of three times. The control group was only given dual-wavelength pulsed dye laser, once a month, a total of three times.Results:Twenty-two cases (73.33%) in the observation group were effective, while 14 cases (46.67%) in the control group were effective. The efficacy of the observation group was better than that of the control group, and the difference was statistically significant (χ 2=4.44, P<0.05). There were no obvious adverse reactions in both groups. Conclusions:Dual-wavelength pulsed dye laser combined with 30% supramolecular salicylic acid is effective and safe in the treatment of moderate and severe facial acne, which is worth popularizing.

Objective:To investigate the clinical manifestation, pathological types, treatment and prognosis of primary tracheobronchial tumors in children.Methods:We retrospectively studied the primary tracheobronchial tumors patients who diagnosed from May 2009 to Jan 2021 in Guangzhou Women and Children Medical Center. The clinical manifestations, pathological types, therapeutic methods and prognosis were analyzed.Results:There were 15 patients identified as the primary tracheobronchial tumors, including synovial sarcoma (1 case), pulmonary inflammatory myofibroblastic tumor(IMT 4 cases), mucoepidermoid carcinoma(7 cases), infantile hemangioma (1 case), Ewing's sarcoma (1 case). Respiratory symptoms are the most complaint at the time of diagnosis including 15 patients with cough, 2 with hemoptysis, and 1 with dyspnea. Endoscopic treatment of tracheobronchial tumors was performed under extracorporeal membrane oxygenation (ECMO) support in 1 patient. Sleeve lobectomy was performed in 3 patients, lobectomies in 6, and local tumor resections in 4 patients including 2 patients suffered second surgery due to tumor recurrence.Conclusion:The clinical manifestations of the primary tracheobronchial tumors in children are nonspecific. Complete resection led to excellent outcome.

[Abstract] Objective: To explore the pathogenosis and prognostic markers for non-smoking female lung cancer patients with bioinfor‐ matics analysis and functional prediction of potential lung cancer associated genes in female non-smokers. Methods: Data for nonsmoking female patients with lung cancer were downloaded from the Gene Expression Omnibus (GEO) database and the differentially expressed genes (DEGs) were identified using GEO2R. DAVID online data base was used to perform gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG), and STRING online software was used to perform protein-protein interaction (PPI) analysis; then the plug-in (M-CODE) was used to screen the key DEGs; finally, GEPIA and Kaplan-Meier plotter were used to perform function prediction and prognosis analysis of key DEGs. Results: A total of 160 DEGs were screened, including 54 up-regulated and 106 down-regulated genes; GO enrichment analysis showed that these DEGs were mainly related to neovascularization, single cell adhesion, positive regulation of GTPase activity and signal transduction (all P<0.05). KEGG pathway analysis revealed that DEGs were mainly involved in cell adhesion molecules (CAMs), leukocyte transendothelial migration, tight junction and endocytosis (all P<0.05); PPI network analysis revealed 8 key DEGs, including TIE1, PECAM1, CLDN5, VEGFD, ICAM2, ESAM, EMCN and ROBO4. Conclusion: TIE1, CLDN5, ICAM2, ESAM, VEGFD and ROBO4 may be the research targets of the pathogenesis of non-smoking female lung cancer patients. PECAM1 and EMCN may be the new bio-markers to predict the progression and prognosis of nonsmoking female lung cancer patients.

Objective To investigate the effect of repetitive transcranial magnetic stimulation (rTMS) on the depressive like behaviors and expression of brain derived neurotrophic factor (BDNF),IL-1β and NF-κB of hippocampal in chronic unpredictable mild stress (CUMS) rats.Methods Thirty-two adult male rats were randomly divided into four groups (n =8):Control group,Control + rTMS group,CUMS group and CUMS + rTMS group.The sucrose preference test,forced swim test and open field test were used to evaluate depressive like behaviors for each groups.In addition,the expression of BDNF,NF-κB and IL-1 β in hippocampal were detected by western blot and ELISA after behavioral test,respectively.Results 1.The effects of rTMS on depressive like behaviors of CUMS rats:in the sucrose preference test,the sucrose preference rate of CUMS rats (0.67 ± 0.06) was significantly lower than Control group (0.91 ± 0.04),which was higher in the CUMS + rTMS group (0.83 ±0.08).In the forced swim test,the immobility time of CUMS group ((26.88 ± 11.33) s) was longer than Control group ((15.22 ± 6.75) s) and CUMS + rTMS group ((18.41 ± 6.95) s).In the open field test,both the total distance travelled and number of central area entry times of CUMS group((849.165 ± 769.01) cm,(7.42 ± 5.68))were significantly shorter ((6224.81 ± 1403.2) cm) and smaller (22.86 ± 3.72) than Control group,and those of the CUMS + rTMS were longer ((4105.57 ± 1516.92)cm) and larger (21.25 ± 3.45).All the behavioral results were statistically significant (P＜ 0.05).And of all the aforementioned behavioral parameters,there were no significant differences between Control group and Control + rTMS group(P＞0.05).2.The effects of rTMS on the hippocampal expressions of BDNF,NF-κB and IL-1β in CUMS rats:compared with Control group,the hippocampal expression of BDNF in CUMS rats was significantly decreased,while the expressions of NF-κB and IL-1β in the hippocampus were significantly increased (P＜ 0.05).Compared with CUMS group,the hippocampa expression of BD-NF in the CUMS + rTMS group was increased,and the expressions of NF-κB and IL-1β in the hippocampus was significantly decreased (P ＜ 0.05).The expressions of BDNF,NF-κB and IL-1β had no differences between Control group and Control + rTMS group.Conclusion rTMS increased the expression of BDNF,reduced the production of NF-κB and IL-1β,and alleviated depressive like behaviors in CUMS rats.

ObjectiveTo explore the relation of heart rate variability (HRV) in neonatal asphyxia with myocardial injury.MethodsContinuous electrocardiographic monitoring by 24-hour Holter recordings was performed in 53 neonates with asphyxia and 40 healthy newborn.The difference of HRV with sinus rhythm was analyzed.Time-domain indexs included standard deviation of R-R intervals (SDNN) ; standard deviation of all mean 5-minute R-R intervals (SDANN) ;standard deviation of all R-R intervals for all 5-minute segments of 24 hours (SDNNindex ) ;root mean squared successive difference (rMSSD) ;percent of NN50 in the total number R-R intervals ( PNN50 ).Results( 1 ) Maximum heart rate,minimum heart rate and average heart rate of 24-hour Holter in healthy newborn were faster than those in newborn with neonatal asphyxia ( P ＜ 0.05 ).And the heart rate was faster in newborn with mild neonatal asphyxia than that in newborn with serious neonatal asphyxia ( P ＜ 0.05 ).(2) SDNN,SDANN of HRV index analysis showed significantly difference between healthy newborn and asphyxia newhom ( P ＜ 0.05 ).There were no difference of SDNN,SDANN,SDNNindex,rMSSD and PNN50 between mild and serious neonatal asphyxia (P ＞ 0.05 ).No significant differences of SDNNindex,rMSSD and PNN50 were found among three groups.ConclusionMyocardial injury caused by neonatal asphyxia can lead to damage of cardiac autonomic nevous and affect heart rate changes.The degree of myocardial injury is related to the degree of neonatal asphyxia.