The synthetic PDCoV N protein gene was optimized and cloned into the pET-30a vector to obtain the pET-30a-N plasmid.Thenthe recombinant plasmid was transformed into three strains of BL21 E.coli using heat-shock to explore protein expression conditions.The expressed proteins was purified using Ni Focurose 6FF(IMAC)and used as antigen to immunize the New Zealand White rabbit to prepare the polyclonal antibody against the PDCoV N protein.The antibody titer was measured by indirect ELISA method.The specificity for the antibody was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the pET-30a-N plasmid showed high expression level in BL21 StarTM(DE 3).The optimal expression condition was 37 ℃ 4 h.The purity of the target protein could reach 90.3％after purification.Indirect ELISA showed that the antibody titers was up to 1∶204 800.Western blot and IFA showed that the produced rabbit polyclonal antibody exhibited good specificity.In conclusion,the polyclonal antibody was prepared which specifically recognized the PDCoV N proteins.The results provided some references for the subsequent exploration of PDCoV N protein function and laid a foundation for establishing a diag-nostic method for PDCoV.

Objective To study the effect and possible mechanism of Siraitiae fructus Qingyan Formula(SQF)on acute pharyngitis(AP)rats induced by ammonia water.Methods The active ingredients and targets of SQF were obtained from TCMSP database,pharyngitis targets were acquired from disease databases such as DrugBank,and the common targets were identified through intersections.Constructing protein-protein interaction(PPI)networks to obtain key targets.GO and KEGG enrichment analysis were carried out,and the"active ingredients-targets-pathways"network was constructed.Discovery Studio 2019 was used for molecular docking of active ingredients.The acute pharyngitis model was induced by fixed point quantitative application of ammonia water.After Siraitiae fructus Qingyan oral thick paste(SQP)low-,medium-and high-dose(4,8,16 g/kg)administration,The body mass,general behavior and symptoms of the rats were monitored continuously,and the score of pharyngeal swelling was recorded;the pathological changes of pharyngeal were observed by HE staining.The classification and the count of inflammatory cells were determined.The expression of inflammatory factors(IL-6,IL-1β,PGE2)and PI3K-AKT signaling pathways in pharyngeal tissues were assessed by quantitative real-time polymerase chain reaction(qRT-PCR).Results A total of 35 active ingredients,223 action targets and 2549 pharyngitis related targets were obtained through network pharmacology,with 153 common targets in total.The key targets were protein kinase B(PKB or AKT).Pathways involved PI3K-AKT signaling pathway.Combined with key targets,it was speculated that PI3K-AKT is an important signaling pathway for the treatment of acute pharyngitis with SQF.Compared with the model group,the related symptoms of AP rats were alleviated after the treatment of SQP.The redness and swelling of pharynx were significantly improved(P<0.001).The hyperplasia of the upper mucosa of pharyngeal was alleviated or disappeared,infiltration of a small amount of inflammatory cells,and hypertrophy and hyperplasia of submucosa glandular cells were alleviated.The numbers of inflammatory cells in blood of rats significantly decreased(P<0.05).And the mRNA expression levels of IL-6,IL-1β and PGE2 in pharyngeal tissues were significantly decreased(P<0.05,P<0.01,P<0.001),while the mRNA expression levels of PI3K,AKT and mTOR mRNA were remarkably raised(P<0.05,P<0.01).Conclusion SQF has obvious improvement effect on AP rats,and its mechanism may be related to reducing inflammation level,improving pharyngeal mucosa hyperplasia,inflammatory cells infiltration,hypertrophy and hyperplasia of submucosal glandular cells,and regulating PI3K-AKT signaling pathway.

This study aims to screen epitope antigens targeting the receptor binding domain(RBD)of porcine epidemic diarrhea virus(PEDV)based on its amino acid sequence(GenBank accession number:AKN45969.1),prepare PEDV RBD polyclonal antibody,and perform their identification.Bioinformatics analysis software was used to predict the potential antigenic epitopes of PEDV RBD and sequence comparison with porcine coronavirus strains was performed,the selected dominant antigen epitopes were then conjugated with keyhole limpet hemocyanin(KLH),to synthesize pep-tides directly and immunize mice to generate specific antibody,Western blot technique and indirect immunofluorescence assay were utilized to identify the specificity of the antibodies,and indirect ELISA method was further applied to determine the antibody potency.Results showed the selected PEDV RBD dominant epitope sequence shared 100％similarity with 18 other PEDV strains,while exhibiting low sequence similarity with 11 TGEV strains(27.8％—29.3％)and 16 PDCoV strains(10.5％—13.4％),indicating good epitope conservation.Western blot showed that the specificity of the prepared peptide antibody specifically recognized the PEDV RED protein overexpressed in Ex-pi293F cells and overexpressed in baculovirus system,and at the same time,the antibody was still able to detect the PEDV S protein expressed in PEDV-infected Vero cells at a 1∶2 000 dilution,while it did not react with TGEV-and PDCoV-infected ST cells,indicating that the good specificity of the peptide antibody.ELISA revealed that the potency of specific antibodies in mouse serum could reach up to 1∶25 600.The above results indicate that bioinformatics techniques were suc-cessfully utilized to predict antigenic epitopes of PEDV RBD protein,and specific PEDV RBD pep-tide antibodies were prepared.

The synthetic PDCoV N protein gene was optimized and cloned into the pET-30a vector to obtain the pET-30a-N plasmid.Thenthe recombinant plasmid was transformed into three strains of BL21 E.coli using heat-shock to explore protein expression conditions.The expressed proteins was purified using Ni Focurose 6FF(IMAC)and used as antigen to immunize the New Zealand White rabbit to prepare the polyclonal antibody against the PDCoV N protein.The antibody titer was measured by indirect ELISA method.The specificity for the antibody was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the pET-30a-N plasmid showed high expression level in BL21 StarTM(DE 3).The optimal expression condition was 37 ℃ 4 h.The purity of the target protein could reach 90.3％after purification.Indirect ELISA showed that the antibody titers was up to 1∶204 800.Western blot and IFA showed that the produced rabbit polyclonal antibody exhibited good specificity.In conclusion,the polyclonal antibody was prepared which specifically recognized the PDCoV N proteins.The results provided some references for the subsequent exploration of PDCoV N protein function and laid a foundation for establishing a diag-nostic method for PDCoV.

Objective To study the effect and possible mechanism of Siraitiae fructus Qingyan Formula(SQF)on acute pharyngitis(AP)rats induced by ammonia water.Methods The active ingredients and targets of SQF were obtained from TCMSP database,pharyngitis targets were acquired from disease databases such as DrugBank,and the common targets were identified through intersections.Constructing protein-protein interaction(PPI)networks to obtain key targets.GO and KEGG enrichment analysis were carried out,and the"active ingredients-targets-pathways"network was constructed.Discovery Studio 2019 was used for molecular docking of active ingredients.The acute pharyngitis model was induced by fixed point quantitative application of ammonia water.After Siraitiae fructus Qingyan oral thick paste(SQP)low-,medium-and high-dose(4,8,16 g/kg)administration,The body mass,general behavior and symptoms of the rats were monitored continuously,and the score of pharyngeal swelling was recorded;the pathological changes of pharyngeal were observed by HE staining.The classification and the count of inflammatory cells were determined.The expression of inflammatory factors(IL-6,IL-1β,PGE2)and PI3K-AKT signaling pathways in pharyngeal tissues were assessed by quantitative real-time polymerase chain reaction(qRT-PCR).Results A total of 35 active ingredients,223 action targets and 2549 pharyngitis related targets were obtained through network pharmacology,with 153 common targets in total.The key targets were protein kinase B(PKB or AKT).Pathways involved PI3K-AKT signaling pathway.Combined with key targets,it was speculated that PI3K-AKT is an important signaling pathway for the treatment of acute pharyngitis with SQF.Compared with the model group,the related symptoms of AP rats were alleviated after the treatment of SQP.The redness and swelling of pharynx were significantly improved(P<0.001).The hyperplasia of the upper mucosa of pharyngeal was alleviated or disappeared,infiltration of a small amount of inflammatory cells,and hypertrophy and hyperplasia of submucosa glandular cells were alleviated.The numbers of inflammatory cells in blood of rats significantly decreased(P<0.05).And the mRNA expression levels of IL-6,IL-1β and PGE2 in pharyngeal tissues were significantly decreased(P<0.05,P<0.01,P<0.001),while the mRNA expression levels of PI3K,AKT and mTOR mRNA were remarkably raised(P<0.05,P<0.01).Conclusion SQF has obvious improvement effect on AP rats,and its mechanism may be related to reducing inflammation level,improving pharyngeal mucosa hyperplasia,inflammatory cells infiltration,hypertrophy and hyperplasia of submucosal glandular cells,and regulating PI3K-AKT signaling pathway.

This study aims to screen epitope antigens targeting the receptor binding domain(RBD)of porcine epidemic diarrhea virus(PEDV)based on its amino acid sequence(GenBank accession number:AKN45969.1),prepare PEDV RBD polyclonal antibody,and perform their identification.Bioinformatics analysis software was used to predict the potential antigenic epitopes of PEDV RBD and sequence comparison with porcine coronavirus strains was performed,the selected dominant antigen epitopes were then conjugated with keyhole limpet hemocyanin(KLH),to synthesize pep-tides directly and immunize mice to generate specific antibody,Western blot technique and indirect immunofluorescence assay were utilized to identify the specificity of the antibodies,and indirect ELISA method was further applied to determine the antibody potency.Results showed the selected PEDV RBD dominant epitope sequence shared 100％similarity with 18 other PEDV strains,while exhibiting low sequence similarity with 11 TGEV strains(27.8％—29.3％)and 16 PDCoV strains(10.5％—13.4％),indicating good epitope conservation.Western blot showed that the specificity of the prepared peptide antibody specifically recognized the PEDV RED protein overexpressed in Ex-pi293F cells and overexpressed in baculovirus system,and at the same time,the antibody was still able to detect the PEDV S protein expressed in PEDV-infected Vero cells at a 1∶2 000 dilution,while it did not react with TGEV-and PDCoV-infected ST cells,indicating that the good specificity of the peptide antibody.ELISA revealed that the potency of specific antibodies in mouse serum could reach up to 1∶25 600.The above results indicate that bioinformatics techniques were suc-cessfully utilized to predict antigenic epitopes of PEDV RBD protein,and specific PEDV RBD pep-tide antibodies were prepared.

Objective To investigate the effect of trunk control on the biomechanical characteristics of lower limb movements during Asian squats(AS)and Western squats(WS)in young adults to provide empirical support for the application and promotion of deep squat training.Methods Twenty-four healthy young male collegiate students performed AS and WS with and without bar control,and their lower limb kinematic and kinetic characteristics were collected using an infrared light-point motion capture system and a three-dimensional(3D)dynamometer.The 3D angles of the lower limbs were obtained using Cortex-642.6.2 software,based on the calculation of Euler angles,and the 3D moments were obtained by applying the inverse dynamics method.The effects of trunk control and deep squatting posture on the lower limb kinematic characteristics were examined using a two-factor analysis of variance with a 2×2 repeated design.Results There was no significant interaction between trunk control and the deep squatting posture for either kinematic or kinetic parameters(P>0.05).The WS group had a large knee flexion angle,peak patellofemoral contact force,and ratio of peak hip and knee extension moments,and small ankle dorsiflexion and hip flexion angles(P<0.05).The deep squat with a bar had a large ankle dorsiflexion angle,peak patellofemoral contact force,and hip flexion angle as well as a small knee flexion angle and ratio of peak hip and knee extension moments(P<0.05).Conclusions WS is helpful for training hip extension muscle groups,whereas AS is helpful for training knee extension muscle strength.The peak patellofemoral joint contact force of the WS is significantly greater than that of the AS;therefore,it is recommended that patients with patellofemoral joint pain use the AS.A squat with a bar can compensate for the body's balance;thus,people with limited ankle dorsiflexion range of motion or anterior tibial muscle weakness may consider trunk control training,such as a deep squat with a bar.This may help improve lower limb stability during squats.

Objective:To analyze the clinical situation of critically ill children with Montgomery T-tube，aiming to summarize the characteristics of T-tube application in pediatric and the experience of postoperative airway management in PICU.Methods:The etiology，clinical characteristics，complications and ICU admissions of patients with Montgomery T-tube admitted to the Pediatric Hospital of Fudan University from April 2019 to December 2021 were analyzed，and the application of T-tube in patients with critical conditions requiring long-term mechanical ventilation was described in the light of clinical experience.Results:During the study period,seven children were admitted to the PICU after T-tube insertion,including three males and four females,aged 9~75 months.Five children received mechanical ventilation.Among them,there were five cases with congenital laryngeal malformations,one case with tracheoesophageal fistula,and one case with laryngeal papilloma.The main complications were sputum blockage,infection,and granulation proliferation.One child died of secretion blockage,while the other children were successfully evacuated from the T-tube.The longest retention time of the T-tube was 367 days.Five patients experienced hoarseness after removing the T-tube,and upon re-examination with fiberoptic bronchoscopy,no recurrence of subglottic stenosis was observed.There was no respiratory distress or wheezing,and there were no abnormalities observed during regular outpatient follow-up after discharge.After discharge,the quality of life of the six surviving children improved compared to preoperative,and they all resumed oral feeding.There were no complaints of swallowing difficulties or aspiration during outpatient follow-up.But they were all combined with malnutrition.Conclusion:The Montgomery T-tube is a secure and dependable airway stent utilized for airway remodeling and the maintenance of airway patency following interventional surgery.For critically ill children,early management of airway clearance and infection prevention are imperative.

OBJECTIVE: To review the current research status and hotspots of acupuncture-assisted tumor chemotherapy and provide references for clinical and basic research in this field. METHODS: The relevant literature on acupuncture-assisted tumor chemotherapy from the inception of the China National Knowledge Infrastructure (CNKI) and Web of Science (WoS) databases to December 10, 2023, was retrieved. CiteSpace6.2.R4 and VOSviewer1.6.20 software was used to analyze publication volume, authors, institutions, source journals, and keywords, etc. and to create visualized mapping. RESULTS: A total of 2 116 articles were included (1 829 from CNKI and 287 from WoS), showing an overall upward trend in publication volume. The scope of acupuncture-assisted tumor chemotherapy has expanded, with a growing variety of research types. In CNKI, the most prolific authors and institutions were from Henan University of CM, forming core research teams. In WoS, the most prolific author was Bao T, and the leading institution was Kyung Hee University in South Korea, although author and institution distribution was more scattered, with close inter-regional collaboration. There were 602 keyword nodes in CNKI and 383 in WoS, with high-frequency keywords in both databases mainly focusing on treatment protocols, cancer types, and chemotherapy-related adverse reactions. CNKI publications highlighted frequent use of acupoints, with more diverse acupoint protocol options, while WoS focused more on different research methods. Recent CNKI studies have focused on improving immune function and quality of life, while "systematic review" emerged as a key term in WoS. CONCLUSION Research on acupuncture-assisted tumor chemotherapy should further strengthen collaboration and communication, focus on improving clinical evidence, and promote wider application of acupuncture in integrative oncology research.
Humans ; Acupuncture Therapy ; Bibliometrics ; Neoplasms/drug therapy* ; Antineoplastic Agents ; China

Objective: To investigate the potential relationship between sensory characteristics and gray matter volumes in children with autism spectrum disorders (ASD), to provide a basis for the diagnosis and treatment of children with ASD. Methods: A total of 40 ASD children who were treated or recovered in Xi an medical institutions and 16 typically developing (TD) children who were from several kindergatens in Xi an were invited for participation. Sensory characteristics were evaluated by the sensory processing and self regulation checklist, 3D structural brain images were obtained with TIWI, and gray matter volumes were analyzed by voxel based morphometry. Sensory characteristics and gray matter volumes were compared between groups and the relationship between sensory characteristics and different gray matter volumes were analyzed. Results: The scores of auditory, visual, tactile, sensory processing ability and sensory under responsivity in the ASD group were lower than those in the TD group ( Z/t =-2.63, -2.57 , -3.11, -2.19, -3.83, P <0.05). Gray matter volumes in nine brain regions increased in the ASD group compared to the TD group, including the left and right posterior inferior lobe, right parahippocamal gyrus, left insula, left media frontal gyrus, left superion occipital gyrus, right superion occipital gyrus, right superion parietal lobe, and right posterion central gyrus ( t =3.53, 3.69 , 3.37, 3.86, 3.61, 3.37, 4.04, 3.38, 3.16, P <0.01). In the ASD group, the scores of visual, vestibular, proprioceptive, sensory processing ability, sensory seeking behavior and sensory over responsivity were negatively correlated with gray matter volumes of left superior occipital gyrus ( r =-0.36, -0.40, -0.39, -0.36, -0.40, -0.36), and the scores of visual, vestibular, and sensory over responsivity were negatively correlated with gray matter volumes of the right superior parietal lobule ( r =-0.36, -0.50, -0.42)( P <0.05). Conclusion The presence of paresthesia in children with ASD is associated with gray matter volumes of the left superior occipital gyrus and right superior parietal lobule.