BACKGROUND:Parkinson's disease has the main pathological changes in the midbrain,especially in the dense substantia nigra,leading to impaired motor and non-motor function in patients.At present,research is limited by cellular heterogeneity,and its pathogenesis still needs to be further elucidated.In recent years,single-cell RNA sequencing(scRNA-seq)has gradually been applied in neurodegenerative diseases,which is of great significance for understanding intercellular heterogeneity,disease development mechanisms,and treatment strategies. OBJECTIVE:To review the research progress of scRNA-seq technology applied to Parkinson's disease in recent years,providing a theoretical basis for the application of scRNA-seq in the treatment and diagnosis of Parkinson's disease. METHODS:The first author used a computer system to search for relevant literature in the CNKI,WanFang,PubMed,and Web of Science databases,with the Chinese search terms"single-cell RNA sequencing,Parkinson's disease,cell heterogeneity,cell subtypes,dopaminergic neurons,glial cells"and English search terms"single-cell RNA seq,Parkinson disease,heterogenicity,subtypes,dopaminergic neurons,glial cells."71 articles were ultimately included for review and analysis. RESULTS AND CONCLUSION:(1)scRNA-seq is a high-throughput experimental technique that utilizes RNA sequencing at the single-cell level to quantify gene expression profiles in specific cell populations,revealing cellular mysteries at the molecular level.Compared with traditional sequencing techniques,scRNA-seq technology is used to reveal the diversity of cell types and changes in specific gene expression in complex tissues under various physiological and pathological conditions through automatic clustering analysis of cell transcriptome.(2)By using scRNA-seq,the development process of dopaminergic neurons and the unique functional characteristics of various cell subtypes are elucidated,in order to better understand potential therapeutic molecular targets.(3)The use of scRNA-seq analysis has improved our understanding of the response of Parkinson's disease glial cells,enabling us to comprehensively map and characterize different cell type populations,identify specific glial cell subpopulations related to neurodegeneration,and draw valuable single cell maps as reference data for future research.(4)The application of scRNA-seq to detect embryonic mice and stem cells will help improve the in vitro differentiation protocol and quality control of cell therapy,as well as evaluate the overall cell quality and developmental stage of dopaminergic neurons derived from stem cells.

OBJECTIVE: To develop a clinical automated diagnostic system for temporomandibular disorders (TMD) based on the diagnostic criteria for TMD (DC/TMD) to assist dentists in making rapid and accurate clinical diagnosis of TMD. METHODS: Clinical and imaging data of 354 patients, who visited the Center for TMD & Orofacial Pain at Peking University Hospital of Stomatology from September 2023 to January 2024, were retrospectively collected. The study developed a clinical automated diagnostic system for TMD using the DC/TMD, built on the. NET Framework platform with branching statements as its internal structure. Further validation of the system on consistency and diagnostic efficacy compared with DC/TMD were also explored. Diagnostic efficacy of the TMD clinical automated diagnostic system for degenerative joint diseases, disc displacement with reduction, disc displacements without reduction with limited mouth opening and disc displacement without reduction without limited mouth opening was evaluated and compared with a specialist in the field of TMD. Accuracy, precision, specificity and the Kappa value were assessed between the TMD clinical automated diagnostic system and the specialist. RESULTS: Diagnoses for various TMD subtypes, including pain-related TMD (arthralgia, myalgia, headache attributed to TMD) and intra-articular TMD (disc displacement with reduction, disc displacement with reduction with intermittent locking, disc displacement without reduction with limited opening, disc displacement without reduction without limited opening, degenerative joint disease and subluxation), using the TMD clinical automated diagnostic system were completely identical to those obtained by the TMD specialist based on DC/TMD. Both the system and the expert showed low sensitivity for diagnosing degenerative joint disease (0.24 and 0.37, respectively), but high specificity (0.96). Both methods achieved high accuracy (> 0.9) for diagnosing disc displacements with reduction and disc displacements without reduction with limited mouth opening. The sensitivity for diagnosing disc displacement without reduction without limited mouth opening was only 0.59 using the automated system, lower than the expert (0.87), while both had high specificity (0.92). The Kappa values for most TMD subtypes were close to 1, except the disc displacement without reduction without limited mouth opening, which had a Kappa value of 0.68. CONCLUSION This study developed and validated a reliable clinical automated diagnostic system for TMD based on DC/TMD. The system is designed to facilitate the rapid and accurate diagnosis and classification of TMD, and is expected to be an important tool in clinical scenarios.
Humans ; Temporomandibular Joint Disorders/diagnosis* ; Retrospective Studies ; Male ; Female ; Adult ; Middle Aged ; Facial Pain/diagnosis* ; Diagnosis, Computer-Assisted/methods* ; Sensitivity and Specificity ; Young Adult

In order to mine candidate vaccine antigens against ticks and to control ticks safely and effectively,the aim of this study was to immunize rabbits with purified aquaporins(AQPs)rD-mAQP1,rDmAQP2 and rDmAQP3 of Dermacentor marginatus.Blood collections for Western blot and ELISA tests were performed.The anti-tick challenge was conducted.The optimal expression conditions of rDmAQP1,rDmAQP2 and rDmAQP3 were screened by SDS-PAGE gel electrophore-sis.The three recombinant proteins were purified by HisSepNi-NTA6FF purification column.Rab-bits were divided into four groups of three rabbits each,including a control group and three immu-nized groups.The three purified recombinant proteins were separately immunized to three groups of rabbits,and the rabbits were immunized once on the 0th,14th and 21st day.Blood samples were collected every 7 days to prepare polyclonal antibodies.The reactivity was detected by Western blot and the antibody titer was detected by ELISA.Tick challenge test was carried out after the anti-body titer increased.The results showed that the optimal expression conditions for rDmAQP1 were induced for 8 h at IPTG concentration of 1.0 mmol/L and 37 ℃;the optimal expression conditions for rDmAQP2 were induced for 7 h at IPTG concentration of 1.0 mmol/L and 37 ℃;and the opti-mal expression conditions for rDmAQP3 were induced for 5 h at IPTG concentration of 1.0 mmol/L and 37 ℃.Western blot results showed that rDmAQP1,rDmAQP2 and rDmAQP3 all had certain reactivity.The ELISA results showed that the antibody titers of rabbits immunized with rD-mAQP1,rDmAQP2 and rDmAQP3 were as follows:the total anti-tick effect of rDmAQP1 protein was 79.74％,and the inhibition rates on average full-blooded tick weight,average egg weight and average egg hatching rate were 9.43％,25.17％and 63.81％,respectively.The total anti-tick effect of rDmAQP2 protein was 78.78％,and the inhibition rates of average full-blooded tick weight,av-erage egg weight and average egg hatching rate of Dermacentor marginatus were 8.30％,20.14％and 68.26％,respectively.The total anti-tick effect of rDmAQP3 protein was 87.91％,and the inhi-bition rates of average full-blooded tick weight,average egg weight and average egg hatching rate were 3.23％,22.47％and 80.5％,respectively.Through serological test and anti-tick test,it has been found that rDmAQP1,rDmAQP2 and rDmAQP3 all have the potential of candidate antigens against ticks,among which rDmAQP3 has the best immune effect,which lays a foundation for the study of the function of rDmAQP1,rDmAQP2 and rDmAQP3.

Peptides that are composed of dextrorotary (d)-amino acids have gained increasing attention as a potential therapeutic class. However, our understanding of the in vivo fate of d-peptides is limited. This highlights the need for whole-body, quantitative tracking of d-peptides to better understand how they interact with the living body. Here, we used mouse models to track the movement of a programmed death-ligand 1 (PD-L1)-targeting d-dodecapeptide antagonist (DPA) using positron emission tomography (PET). More specifically, we profiled the metabolic routes of [⁶⁴Cu]DPA and investigated the tumor engagement of [⁶⁴Cu/⁶⁸Ga]DPA in mouse models. Our results revealed that intact [⁶⁴Cu/⁶⁸Ga]DPA was primarily eliminated by the kidneys and had a notable accumulation in tumors. Moreover, a single dose of [⁶⁴Cu]DPA effectively delayed tumor growth and improved the survival of mice. Collectively, these results not only deepen our knowledge of the in vivo fate of d-peptides, but also underscore the utility of d-peptides as radiopharmaceuticals.

Objective:To provide experimental evidence for genetic counseling and prenatal diagnosis by analyzing the clinical characteristics, screening and identification of the function of suspicious variants in a X-1inked spondyloepiphyseal dysplasia tarda (SEDT) family.Methods:The family members' medical history, general physical examination, femur, spine X-ray examination were collected. Peripheral blood samples of the family members were collected and DNA was extracted from these samples. Sequencing clinical whole exons of proband DNA by targeted gene high-throughput sequencing method, then analysis sequencing data. The suspicious mutation was confirmed in pedigree members by PCR and Sanger sequencing. Reverse transcription polymerase chain reaction (RT-PCR) experiments of total RNA from blood lymphocytes were performed. The amplification of exons 3 and 4 of the pathogenic gene were amplified and identified by agarose gel. The expression of the pathogenic gene was also detected.Results:Three affected males of the family were diagnosed with SEDT according to their clinical and radiological features. A nonsense mutation in the transport protein particle complex subunit 2 ( TRAPPC2) gene NM_001011658: c.91A>T (p.K31*) was found in the proband using whole exome sequencing. This variation was also detected in his cousin, but not in non-phenotypic members of the family. The RT-PCR result for amplification of exon 3 and 4 of peripheral blood lymphocytes was the same as those of normal controls, indicating that the mutation did not affect the splicing of transcripts. qPCR results showed that the transcriptional expression of TRAPPC2 in patients was significantly lower than that in family normal controls and normal people controls. Conclusion:Identification of the novel nonsense mutation (c.91A>T) in the SEDT family enables early patients screening, carrier detection, genetic counseling, prenatal diagnosis, and clinical prevention and treatment. The detailed genotype/phenotype descriptions contribute to the SEDT mutation spectrum. The study of the function of TRAPPC2 mutation will help to further elucidate the role of sedlin in cartilage.

Objective To observe the effect of variable frequency vibration therapy while sitting in an anti-spasmodic posture on spasticity and the motor function of the upper limbs among stroke survivors.Methods Thirty stroke survivors with upper limb spasticity were randomly divided into a treatment group and a control group,each of 15.Both groups were given routine rehabilitation training for 4 weeks while the treatment group was additionally provided with variable frequency vibration training while sitting in anti-spasmodic postures.Before and after the treatment,the modified Ashworth scale (MAS) was used to assess spasticity.The root mean square (RMS) value of the surface electromyogram amplitude of the affected biceps when extended passively and those of the triceps,obliques and multifidus in maximum isometric contraction was measured and recorded.The motor function of the affected upper limbs was evaluated using the active range of motion (A-ROM) of the shoulder,elbow and wrist,as well as a Fugl-Meyer assessment (FMA).Moreover,ability in the activities of daily living (ADL) was assessed using the modified Barthel index (MBI).Results After the treatment,significant improvement was observed in the average MAS,A-ROM,RMS,FMA and MBI results in both groups compared to those before the treatment.Moreover,the results in the treatment group were significantly better than those of the control group,on average.Conclusions Variable frequency vibration therapy while sitting in an anti-spasmodic posture combined with traditional rehabilitation is more effective than the latter alone in relieving spasticity as well as improving motor function and ability in the activities of daily living among stroke survivors with the upper limb spasticity.

Hospital image information system is evolving from department-oriented to patient-oriented. This paper introduces the implementation of Beijing University People's Hospital image data center. The ideas, approaches and the technological standards for image data center's system integration are proposed. Additionally, the problems in practical application and the outlook for this system are discussed.

ObjectiveTo assess the effects of crude extract of Cape Jasmine in the experimental Alzheimer's model induced by Aβ25-35 and the memory acquisition impaied model induced by ibotenic acid(IBO).MethodsAlzheimer's dementia of mice was induced by Aβ25-35 i.cv. treatment and the impaired memory acquisition model was induced by IBO injected into the forebrain nucleus basalis. The effects of crude extract of Cape Jasmine on the learning and memory function of model animals were evaluated with Morris water maze and step-through test in 3 dose groups(12-5, 25, 50 mg/kg). Donepezil(0-75 mg/kg)was used in the positive control group.ResultsMorris water maze test showed that the spatial learning and memory ability of the model mice significantly improved in three crude extract of Cape Jasmine groups(P<0-05). Meanwhile, the impaired function of the rats induced by IBO significantly improved in the medium dose group(P<0-01). The electric shock latent period in step-through box test prolonged and the frequency of electric shock decreased within 3 minutes in three groups.ConclusionCrude extract of Cape Jasmine can improve the learning and memory function of mice or rat in the model group.

ObjectiveTo explore the possible mechanisms of Wuling Jun Power by the DNA microarray technique.MethodsAn experimental depression model was established by exposing the mouse to a chronic mild stress procedure. The total RNA was extracted reverse-transcripted and hybrided to the mouse 1-2 cDNA microarray (Clontech). The difference of expression profiles between control model, Wuling Jun powder and fluoxetine-treated groups were analyzed by the Image 2-1 Software.Results130 genes were significantly altered in stress group compared with the control groups. Among them, 116 genes were up-regulated and 14 genes were down-regulated. Meanwhile, 85 genes significantly changed in the Wuling Jun powder treated group with 34 genes up-regulated and 51 genes down-regulated compared with the model groups. For the Fluoxetine-treated group, 133 genes significantly changed with 35 genes up-regulated and 98 genes down-regulated compared with the model groups. These genes were associated with many aspects of life including receptor activity, protein kinases, inflammatory factors, transferrin, neurogenesis and so on.ConclusionMultiple genes were affected by the stress exposure. Altered changes of some genes were normalized by Wuling Jun powder and Fluoxetine treatment. In general, the mechanisms of Wuling Jun powder and Fluoxetine are similar, but also with minor difference.

ObjectiveTo investigate the effects of Jiuqiang Naoliqing(JNQ) on the morphology of the vital organs(the brain,kidney and heart) in acute hypertension rats.MethodsThe model was established by L-NG-nitro arginine(L-NNA),which can inhibit the synthesis of nitric oxide(NO).Affected animals were administrated with high,medium or low dose of JNQ while nifedipine was used as the positive control.Effects on cardiac function and morphology of the vital organs were investigated.Results2 weeks after the establishment of the model,the affected rats had a 36% higher blood pressure compared with the control group.The acute hypertension rats showed significant pathologic changed in the brain,kidney and heart,while there was no obvious difference in the heart rate,electrocardiaogram,and blood pressure between the administrated groups.After treatment with JNQ,the contraction force and the oxygen extraction of the myocardiocytes were significantly reduced and the increasing velocity of the left ventricular pressure was decreased.The brain,kidney and heart showed improvement in pathology analysis to different levels.ConclusionAlthough it has no obvious effects on the heart rate and blood pressure in acute hypertension rats,JNQ can decrease the oxygen extraction in myocardiocytes and can obviously alleviate brains,kidneys and hearts pathologic changes.