Objective: To construct gene recombinant expression plasmids of human anti-P antibody with IgG1 and kappa chain based on the human hybridoma cell line. Methods: Starting from the specific RT-PCR products encoding the variable regions of the IgH chain and IgL chain of the anti-P monoclonal cell line, appropriate restriction enzyme digestion sites were introduced at both ends of the VH and VL fragments through nested PCR. The plasmids carrying the antibody constant region and the nested PCR products of VH and VL were ligated by the action of T4 ligase and subsequently transferred into competent E. coli DH5ɑ, and positive clones were selected in the antibiotic resistant LB medium. After sequence confirmation, recombinant plasmids DNA were transfected into HEK293T cells, and the recombinant antibody obtained in the culture supernatant. The characteristics of recombinant expression antibodies were determined by using rapid antibody isotying kit, serological agglutination tests and flow cytometry. Results: Recombinant gene expression vectors, pFUSEss-CHIg-hG1+VH, pFUSE2ss-CLIg-hκ+VL, were successfully constructed. Human IgG1 kappa light chain antibodies were detected in the supernatant of HEK293T cells transient transfected with recombinant plasmids. After the supernatant was ultra-filtered and concentrated, it could cause agglutination reactions with P antigen-positive red blood cells. The mean fluorescence intensity (MFI) of the reaction between recombinant antibodies and antigen-positive red blood cells in flow cytometry experiments was higher than that of antigen-negative red blood cells. Conclusion: The experimental study on the conversion of red blood group antibody types by genetic engineering technology represents a beneficial exploration towards establishing a feasible technical route for the development of genetic recombination and modification of antibodies reagent.

Objective To explore the syndrome differentiation rules of the Yin Dan and Yang Dan method,and summarize Professor Yang Zhimin's valuable experience in the treatment of insomnia.Methods The case data of 11390 insomnia patients treated by Professor Yang Zhimin was retrospectively analyzed,and the distribution of symptoms,signs,pathogenesis and syndrome differentiation rules of insomnia patients was examined based on latent structure analysis.Result A total of 44 symptoms and signs can explain the overall situation of insomnia patients,showing the characteristics of both excess and deficiency,cold and heat.A total of 36 hidden categories(symptom clusters and pathogenesis)was composed.Symptoms and signs such as fear of wind and cold can explain the overall situation of insomnia patients with cold syndrome due to weakness of Wei Qi,symptoms and signs such as pale eyelid can explain the overall situation of insomnia patients with heat syndrome due to weakness of Ying Xve,and established diagnostic criteria for insomnia patients with cold syndrome due to weakness of Wei Qi and heat syndrome due to weakness of Ying Xve.Conclusion Based on the Yang Dan and Yin Dan method to distinguish and treat insomnia from the perspective of disharmony between Ying and Wei,it is in line with the basic understanding of the pathogenesis of insomnia in the classics of traditional Chinese medicine.The pattern differentiation rules obtained through the latent structure analysis method are helpful to summarize valuable clinical experience and provide a certain reference and basis for clinical practice.

The global incidence of head and neck cancer (HNC) is rising, with over 60% of patients presenting at a locally advanced stage. Radiotherapy remains a cornerstone of HNC treatment, and advancements in modern techniques and concurrent chemotherapy have improved local control and survival rates of HNC patients. However, these benefits also bring challenges in the management of toxicities. Due to the proximity of salivary glands and tumors, especially the highly radiosensitive parotid and submandibular glands, this condition is among the most common adverse effects of radiotherapy. Radiation damages acinar cells and ducts, causing glandular atrophy, fibrosis, and reduced saliva secretion, thereby leading to xerostomia and related complications. The risk and severity of injury are associated with the radiation dose and volume affecting the glands. Prevention and management strategies emphasize precise radiotherapy planning, target optimization, and supportive care. Emerging multimodal imaging techniques offer potential for non-invasive prediction and early diagnosis and treatment of radiation-induced salivary gland injury. Future research in regenerative medicine, tissue engineering, and molecular biology aims to elucidate molecular mechanisms, such as signaling pathways and genomics, facilitating personalized strategies to mitigate radiotherapy-induced toxicities and enhance the quality of life of patients.

Objective:To explore the effective active components of Qi Bi Anshen decoction(QAT)in the treatment of autism spectrum disorder(ASD)and its effects on ASD behaviors and related lipid metabolism abnormalities.Methods:24 adult male Sprague-Dawley rats were randomly divided into 3 groups:Control group,PPA group and PPA+QAT group.The ASD rat model was established by intracerebroventricular injection of propionic acid,and the QAT administration group was given intragastric administration for 7 days.Behavioral tests were conducted to detect the so-cial,repetitive stereotyped and anxiety-like behaviors of rats.UPLC-MS was used to analyze the differential metabolites and enriched pathways of rats.Network pharmacology was used to screen the effective active monomer components of QAT involved in regulating ASD.10 sexually mature C57BL/6J mice were randomly paired in male-female cages.The ASD mouse model was established by a single intraperitoneal injection of sodium valproate to pregnant mice.The preg-nant mice were randomly divided into 3 groups:Control group,VPA group and VPA+QUE group.The administration group was given QUE in the drinking water of pregnant mice until the end of the perinatal period.Behavioral tests were conducted to detect ASD-like behaviors in mice.Reagent kits were used to detect the contents of alkaline phosphatase(AKP),alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglycerides(TG)and total cholesterol(TC)in the liver and serum of mice.Oil red O staining was used to observe the morphology of liver cells.Results:QAT administration could improve the ASD-like behaviors induced by PPA(P＜0.05).UPLC-MS analysis showed that the differential metabolites of each group of rats were mainly enriched in lipid metabolism pathways.Network pharmacol-ogy screening identified QUE as the effective active monomer component.QUE administration could improve the ASD-like behaviors induced by VPA(P＜0.05).QUE administration could reverse the abnormal changes in AKP,TC and TG in the liver induced by VPA(P＜0.05)and reduce lipid droplet deposition in the liver.Conclusion:The active monomer component QUE in QAT has therapeutic effects on ASD behaviors and related liver lipid metabolism abnormali-ties.

Angiotensin-converting enzyme 2 (ACE2) is known for its role in regulating blood pressure and electrolyte balance, but it has also been identified as the functional receptor for coronaviruses.With the global spread of corona virus disease 2019, the role of ACE2 across different age groups, particularly in children, has garnered increasing attention.Studies indicate that the distribution of ACE2 in the respiratory systems of children and adults differs significantly, which implies a distinct functional role in pediatric respiratory diseases.This paper reviews the latest research progress on ACE2 in children′s diseases such as corona virus disease 2019, acute respiratory distress syndrome, community-acquired pneumonia, and asthma.

Objective To explore the syndrome differentiation rules of the Yin Dan and Yang Dan method,and summarize Professor Yang Zhimin's valuable experience in the treatment of insomnia.Methods The case data of 11390 insomnia patients treated by Professor Yang Zhimin was retrospectively analyzed,and the distribution of symptoms,signs,pathogenesis and syndrome differentiation rules of insomnia patients was examined based on latent structure analysis.Result A total of 44 symptoms and signs can explain the overall situation of insomnia patients,showing the characteristics of both excess and deficiency,cold and heat.A total of 36 hidden categories(symptom clusters and pathogenesis)was composed.Symptoms and signs such as fear of wind and cold can explain the overall situation of insomnia patients with cold syndrome due to weakness of Wei Qi,symptoms and signs such as pale eyelid can explain the overall situation of insomnia patients with heat syndrome due to weakness of Ying Xve,and established diagnostic criteria for insomnia patients with cold syndrome due to weakness of Wei Qi and heat syndrome due to weakness of Ying Xve.Conclusion Based on the Yang Dan and Yin Dan method to distinguish and treat insomnia from the perspective of disharmony between Ying and Wei,it is in line with the basic understanding of the pathogenesis of insomnia in the classics of traditional Chinese medicine.The pattern differentiation rules obtained through the latent structure analysis method are helpful to summarize valuable clinical experience and provide a certain reference and basis for clinical practice.

Objective To observe the correlations between left atrial myocardial strain and left ventricular function in children with Duchenne muscular dystrophy(DMD).Methods Fifty-one DMD children(DMD group)and 42 healthy one(control group)were prospectively enrolled.The parameters of routine ultrasound and three-dimensional speckle-tracking echocardiography(3D-STE)were compared between groups,and the correlations between left atrial strain parameters and left ventricular function parameters were analyzed.Results The mitral annular lateral wall velocity(e'),left ventricular ejection fraction(LVEF),left atrial ejection fraction(LAEF),left atrioventricular coupling index(LACI),left ventricular global longitudinal strain(LVGLS),left atrial strain during reservoir phase(LASr)and left atrial strain during conduit phase(LAScd)were all lower,while mitral early diastolic peak flow velocity/e'(E/e'),left atrial stiffness index(LASI)and left atrial filling index(LAFI)were higher in DMD group than those in control group(all P＜0.05).In DMD group,LAEF,LASr and LAScd were moderately positively correlated with LVGLS(r=0.409,0.437,0.440,all P＜0.05),LAFI and LASI were weakly negatively correlated with LVGLS(r=-0.207,-0.223,both P＜0.05),while LASr was moderately positively correlated with e'(r=0.419,P＜0.05).Conclusion The left atrial myocardial strain was correlated with left ventricular systolic and diastolic function in DMD children.

Objective To systematically investigate the impact of music intervention on neuroplasticity in patients with Alzheimer's disease(AD).Methods PubMed,EBSCOhost,Web of Science,Embase,CNKI,Wanfang data and VIP were searched for studies pub-lished up to February,2025.They were analysed using GingerALE 3.0.2 for meta-analysis.Results Five fMRI studies were included,involving 134 participants(99 with AD).A total of 49 activation coordinates were extracted.The main activation regions in the healthy group were concentrated in the left precuneus,the right inferior frontal gyrus and the right superior temporal gyrus,while the primary activated regions in AD group shifted to the right operculum and adjacent right insula,left superior frontal gyrus,right medial frontal gyrus,and left superior temporal gyrus,indicating that during music processing,neural activation patterns in AD patients shifted from higher-order associative cortices to subcortical structures,accompanied by compensatory changes in frontal lobe activation and a loss of activation in the parietal lobe,particularly in the precuneus.Conclusion Music intervention can activate the relatively preserved subcortical-limbic system in patients with AD and promote compensatory reorganization of the frontal lobes.

Bile acids encompass primary bile acids and secondary bile acids,the latter of which are produced from primary bile acids in the gut.In recent years,bile acids,compounds with a steroid structure found in bile,have been shown through extensive clinical investigations and fundamental studies to play a significant role in regulating intestinal and systemic immune homeostasis,inflammatory responses,and neurological disorders.In the gut,primary and secondary bile acids can disrupt the invasion of pathogenic microorganisms into the host due to their unique physical properties.Additional-ly,they regulate downstream signaling pathways by binding to effector receptors,thereby performing vital biological func-tions,including the modulation of immune responses.Consequently,bile acids function as essential small molecules in the pathophysiological processes of various intestinal diseases and extra-intestinal tissue conditions influenced by intestinal homeostasis.This article focuses on the role and regulatory mechanisms of the"microbe-bile acid"axis in intestinal inflam-mation,elucidating the synthesis and metabolism of bile acids and the significance of intestinal microbes in signal trans-duction via bile acids during the inflammatory process.It aims to provide a foundation and reference for the development of drugs and therapeutic strategies targeting diseases related to intestinal inflammation and immune dysregulation.

OBJECTIVE To explore the effect of fluconazole on proteomics of Trichosporon asahii so as to reveal the responding process of T.asahii to fluconazole stress and the resistance mechanisms to azoles on the protein level.METHODS T.asahii AS 2.2174 was chosen as the research subject,the minimum inhibitory concentration(MIC)of fluconazole was determined by broth microdilution assay.The protein abundance of T.asahii was detec-ted by means of tandem mass tag(TMT)technique combined with liquid chromatography-tandem mass spectrom-etry(LC-MS/MS)before and after the treatment with fluconazole(1× MIC).The differentially expressed pro-teins(DEPs)were identified based on the screening standards of fold change ≥1.20 or ≤0.83 and P＜0.05.Gene ontlogy(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis were performed for the DEPs so as to understand the biological property of the DEPs and the major biological pathways that the DEPs in-volved in.Finally,the targeted validation was carried out for the targeted differentially expressed proteins by using multiple reaction monitoring(MRM).RESULTS The MIC of fluconazole to T.asahii AS 2.2174 was 8 μg/ml.Totally 196 DEPs were identified,including 93 upregulated DEPs and 103 downregulated DEPs.The function en-richment analysis showed that the DEPs mainly participated in synthesis and metabolism of sterols,drug metabo-lism,stress response,energy metabolism and intertranslation.The targeted DEPs showed the consistent expres-sion trends in MRM target validation and TMT-LC-MS/MS.CONCLUSIONS The protein abundance of T.asahii has remarkable change under the fluconazole stress.The bioinformatics analysis reveals the complicated molecular mechanisms of T.asahii in response to the fluconazole stress,which may offer valuable ideas for understanding the drug resistance to azoles and developing new drug targets.