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.

ObjectiveTo explore the effects of exosomal CXCL on the biological behavior of cervical cancer cells and its underlying mechanisms. MethodsChemokine CXCL was first screened through bioinformatics databases. The GEPIA database was analyze CXCL expression in cervical cancer tissues and adjacent normal cervical tissues. Western blot was performed to detect CXCL expression levels in cervical cancer cells （Caski） and normal cervical epithelial cells （H8）. The successful isolation of exosomes was confirmed by nanoparticle tracking analysis， transmission electron microscopy （TEM） and Western blot. ELISA was employed to detect the expression level of exosomes CXCL was determined by ELISA. After CXCL knockdown via siRNA transfection， cells were divided into three groups： blank control， negative control and experimental groups. Cell proliferation was evaluated using the CCK-8 assay， while cell migration and invasion were assessed by Transwell assays. ResultsExosomal CXCL expression was significantly upregulated in cervical cancer cells compared with normal cervical epithelial cells （P<0. 01）， and also markedly elevated in cervical cancer tissues compared with adjacent normal tissues. After low expression of CXCL knockdown significantly reduced CXCL expression in both cancer cells and their derived exosomes（P<0. 05）. Low expression markedly inhibited the proliferation， invasion and migration abilities ConclusionSilencing exosomal CXCL may inhibit the malignant biological behavior of cancer cells.

Exosomes are small vesicles with a lipid bilayer membrane structure that have applied in precision medicine due to their non-invasive nature， high accessibility， and stability. Exosomes play a crucial role in processes such as tumor metastasis， invasion， and angiogenesis. Gynecological malignancies primarily include cervical cancer， ovarian cancer， and endometrial cancer， and their early diagnosis and treatment have long been a focus of research. As novel biological markers， exosomes exhibit high specificity and can effectively block the occurrence and progression of gynecological malignancies. This article explores the diagnostic and therapeutic applications of exosomes in cervical cancer， ovarian cancer， and endometrial cancer in detail. In cervical cancer， exosomes are involved in processes such as HPV infection， angiogenesis， and immune evasion， with specific miRNAs （such as miR-30d-5p and let-7d-3p） serving as diagnostic markers. Furthermore， exosomes can act as targeted drug delivery vehicles and vaccine development platforms. In ovarian cancer， the miRNAs carried by exosomes （such as miR-21 and the miR-200 family） have reference value for early diagnosis， and exosomes play an important role in chemotherapy resistance and tumor progression. For endometrial cancer， miRNAs in exosomes （such as miR-15a-5p and miR-106b-5p） can serve as biomarkers for early detection. Additionally， this article highlights the challenges faced by exosomes in clinical applications， such as the complexity of isolation and extraction and the identification of cell sources， and emphasizes the necessity for further basic research and clinical trials. This study provides new ideas and methods for the early diagnosis and precision treatment of gynecological malignancies， holding significant theoretical and clinical importance.

Objective To track and evaluate the implementation and application of the occupational health standard Radiation shielding requirements for radiotherapy room—Part 4: Radiotherapy room of ²⁵²Cf neutron afterloading (GBZ/T 201.4-2015) by radiation health technical service agencies, medical institutions, health supervision agencies, and radiotherapy facility design units, and to provide a scientific basis for the further revision and implementation of this standard. Methods Following the Guideline for health standards tracking evaluation (WS/T 536-2017) and the project implementation plan, relevant practitioners were randomly selected for a questionnaire survey. The survey primarily focused on their awareness, standard training, application, and revision suggestions of GBZ/T 201.4-2015. The results were summarized and analyzed. Results A total of 168 evaluation questionnaires were collected from relevant practitioners in 28 provinces. Only 31.6% of the respondents reported being “well familiar” or “ familiar” with the standard, 27.4% of the respondents believed that the standard was widely used, and 45.2% of the respondents believed that the standard could meet the needs of their work. Only 14.9% of the respondents had received relevant training on the standard, more than half of the respondents had not applied the standard within the past 10 years, and 45.2% of the respondents believed that the standard "needs to be revised". Conclusion Due to the small number of californium-252 neutron afterloading radiotherapy devices in operation on the market, the overall awareness of the standard is low, suggesting that relevant authorities need to strengthen training and publicity of the standard, and that certain sections of the standard need to be revised or merged.

Objective To analyze the risk factors,traditional Chinese medicine(TCM)syndromes,and pathogen distribution in patients with bronchiectasis(BE)complicated with type 2 diabetes mellitus(T2DM).Methods From June 2022 to June 2024,a total of 299 patients with acute exacerbation of BE admitted to Guangdong Integrated Chinese and Western Medicine Hospital Affiliated to Guangzhou University of Chinese Medicine were selected.Based on the presence of T2DM,the patients were divided into the BE-T2DM group(74 cases)and the BE-only group(225 cases).Clinical data of the patients were collected,and univariate analysis and multivariate logistic regression models were used to identify risk factors for BE complicated with T2DM.TCM syndromes and pathogen distribution were statistically analyzed.Results(1)Univariate analysis showed that there were statistically significant differences between the two groups in gender,age,body mass index(BMI),hypertension,coronary atherosclerotic heart disease(shortened to coronary heart disease),atherosclerosis,the ratio of forced expiratory volume in one second to forced vital capacity(FEV1/FVC),white blood cell count(WBC),neutrophil count analysis for the risk factors showed that gender,BMI,hypertension,and FEV1/FVC were the independent risk factors for BE complicated with T2DM.(2)In terms of the distribution of TCM syndromes,both groups were mainly characterized by phlegm-heat accumulating in the lung syndrome and phlegm-damp obstructing the lung syndrome,and BE-T2DM group had a higher proportion of phlegm-heat accumulating in the lung syndrome.(3)For the infection of pathogens,BE-T2DM group had a higher infection rate of Haemophilus influenzae,Acinetobacter baumannii,and Klebsiella pneumoniae,while the BE-only group was predominantly infected with Pseudomonas aeruginosa;BE-T2DM group had a significantly higher rate of viral infections,mainly infected with influenza A virus,rhinovirus,and SARS-CoV-2;BE-T2DM group also suffered from fungal infections,usually infected with Candida albicans.Conclusion For BE patients complicated with T2DM,the independent risk factors are gender,BMI,hypertension,and FEV1/FVC;the common TCM syndromes are phlegm-heat accumulating in the lung and phlegm-damp obstructing the lung;pathogen infections are mainly caused by Gram-negative bacteria,viruses,and fungi.

Objective To investigate the effects of high-altitude hypoxic environment on the pharmacokinetic characteristics and brain tissue distribution of phenytoin sodium in epileptic rats.Methods A total of 70 male SPF-grade Wistar rats aged 2 months and weighing(200±20)g were used in the study.An epilepsy model was induced in the rats using the lithium chloride-pilocarpine method.The successfully modeled rats were randomly assigned to a normoxic treatment group and a high-altitude hypoxic treatment group.Phenytoin sodium was administered via intragastric gavage at a dose of 50 mg/kg in both groups.Blood samples were collected from the orbital venous plexus before treatment and 0.5,1,2,3,4,6,8,10,and 24 h post treatment.The animals were euthanized after the final blood collection,and samples of the liver and the whole brain tissue were collected.In the brain tissue distribution experiment,brain tissue samples were collected at 0.5,1,2,and 4 h after drug administration.The concentration of phenytoin sodium in rat plasma and brain tissue was determined by liquid chromatography-tandem mass spectrometry(LC-MS/MS),and the pharmacokinetic parameters were calculated using WinNolin 8.1 software.The expression levels of CYP2C9 in liver tissue and those of P-gp in brain tissue of epileptic rats were determined by Western blot.Results Compared with those in the normoxia group,the peak concentration,peak time,and half-life of phenytoin sodium in the high-altitude hypoxia group were significantly decreased by 46.0%,42.3%,and 55.5%,respectively(all P<0.05);the clearance rate was significantly increased by 162.0%(P<0.05);and the area under the curve of plasma concentration-time curve was decreased by 45.6%(P<0.01).At 0.5,1,and 2 hours after administration,compared with that in the normoxia treatment group,the concentration of phenytoin sodium in the brain tissue of the high-altitude hypoxia treatment group was significantly decreased by 78.1%,63.5%,and 32.5%,respectively(all P<0.05).Western blot results showed that the expression levels of CYP2C9 in the liver tissue and P-gp in the brain tissue of rats in the high-altitude hypoxia group were approximately 1.78 and 1.65 times higher than those in the normoxia group,respectively(both P<0.05).Conclusion The hypoxic environment at high altitudes can promote the metabolism of phenytoin sodium,reduce its absorption efficiency,and change the characteristics its distribution in the brain,which may be related to the up-regulation of the expression of CYP2C9 in the liver and that of P-gp in the brain.

Objective To investigate the changes of OAT expression in bone cells in chronic renal failure(CRF)and involved mechanism,and to explore the effect of OAT expression on bone metabolism.Methods Ra-ndomly divide the rats into a control group(n=6)and a model group(n=6).The model group established a rat chronic renal failure model using"single nephrectomy+adenine gavage"method,and the red blood cell(RBC)and hemoglobin(HGB)of the rat body were measured using a blood routine analyzer;Measure indicators such as creatinine(Cr),urea nitrogen(BUN),uric acid(UA),blood calcium(Ca2+),and blood phosphorus(P3+)using a fully automated biochemical analyzer;Pathological examination of rat kidneys;X-ray examination of rat tibia;Immunohistochemical examination of bone tissue OAT1 level.Results The bone density of the model group rats is lower than that of the control group;The calcium and phosphorus metabolism of rats in the model group was in metabolic disorder,and the OAT1 value of bone tissue binding was much lower than that of the control group(P= 0.0018),which was statistically significant.(P=0.0018)Conclusion Chronic renal failure affected the binding ability of OAT1 in bone tissue,leading to the metabolic disorder for calcium absorption and phosphorus metabolism,thus aggravating renal osteodystrophy(P<0.05).

BACKGROUND:The researchers noted that upon embedding clinical-grade catgut and polyglycolide-co-lactide threads in the normal human"Zusanli"(ST 36)acupoint,the local area displayed temporal and inflammatory stimulatory effects,resulting in thread differentiation.However,the underlying mechanism behind thread involvement remains to be studied. OBJECTIVE:To investigate the expression levels of calcitonin gene-related peptide,5-hydroxytryptamine,leukotriene B4,and bradykinin at point"Zusanli"(ST 36)in rats after embedding catgut and polyglycolide-co-lactide respectively at different time points. METHODS:110 male SD rats were divided into a blank group(10 rats),a catgut embedding group(50 rats),and a polyglycolide-co-lactide embedding group(50 rats)according to the random number table method.In the blank group,no thread was embedded.In catgut embedding group and the polyglycolide-co-lactide embedding group,the thread was embedded in the left side of the ST36 acupoint once.Tissue was collected from the left side of the ST36 acupoint area 8 hours,3,7,14,and 21 days after embedding.The expression levels of calcitonin gene-related peptide and 5-hydroxytryptamine were detected by immunohistochemistry,and the contents of leukotriene B4 and bradykinin were detected by ELISA. RESULTS AND CONCLUSION:(1)Compared with the blank group,the expression of calcitonin gene-related peptide,5-hydroxytryptamine,bradykinin,and leukotriene B4 was significantly increased in the 8 hours,3,7,14,and 21 days of the catgut embedding group(P<0.05);calcitonin gene-related peptide expression was significantly increased in 8 hours,3,7,and 14 days in the polyglycolide-co-lactide embedding group(P<0.05);the expression of bradykinin was significantly increased in 8 hours,3,and 7 days(P<0.05);the expression of leukotriene B4 was significantly increased at 8 hours,3,7,14,and 21 days(P<0.05).(2)Compared with the polyglycolide-co-lactide embedding group,the expression of calcitonin gene-related peptide was increased at 7,14,21 days after thread embedding(P<0.05),and the expression of 5-hydroxytryptamine was increased at 8 hours,3,7,14 and 21 days after thread embedding(P<0.05);contents of leukotriene B4 and bradykinin in tissues were increased at 8 hours,3,14 and 21 days after embedding(P<0.05)in the catgut embedding group.(3)The results show that calcitonin gene-related peptide,5-hydroxytryptamine,leukotriene B4,and bradykinin in the acupoint region alter after catgut embedding in the ST36 of rats,as well as the alteration of calcitonin gene-related peptide,leukotriene B4,and bradykinin is found in the acupoint region after polyglycolide-co-lactide embedding in rats,which may be one of the mechanisms involved in the local time sensitive stimulus effects caused by embedding threads at acupoints.Moreover,there is a discernible difference between the two thread types.

Objective:To investigate the application value of modified multivisceral trans-plantation (MMT) in chronic intestinal pseudo-obstruction (CIPO) secondary to autoimmune enteril leiomyositis (AEL).Methods:The retrospective and descriptive study was conducted. The clinico-pathological data of a recipient who was admitted to Beijing Tsinghua Changgung Hospital Affiliated to Tsinghua University on February 2022 and underwent MTT for CIPO secondary to AEL were collected. The recipient was a male, aged 29 years old. Results of preoperative histopathological examination showed that there were muscle plexus and ganglion cells in the rectum, sigmoid colon, ascending colon, intrinsic muscle layer of ileum, and a small amount of submucosal layer. There was also a small amount of chronic inflammatory cell infiltration in the muscle, indicating a high possi-bility of diagnosis of neurogenic CIPO.Results:(1) Surgical situations. The operation time was 14 hours and 30 minutes, and the cold ischemia time was 9 hours and 30 minutes. The intra-operative blood product dosage included 14 U of red blood cells, 1 400 mL of fresh frozen plasma, and two therapeutic doses of platelets. (2) Postoperative histopathological examination. Results of postoperative histopathological examination showed chronic inflammation and local erosion of the small intestine and duodenal mucosa, with scattered disappearance of the focal mucosal muscle layer; There is a large infiltration of CD3 + and CD8 + lymphocytes in the lamina propria, especially in the muscularis propria. In severe lesions, there is infiltration of ribbon lymphocytes in the subserosal and muscular layers; Muscle fiber degeneration, reduction, and fibrosis. Deposition of pigment granules in the cytoplasm of smooth muscle cells; No abnormalities were found in the intermuscular, submu-cosal ganglia, and Cajal cells; Fibrosis of the serosal layer with local cellulose exudation; Chronic inflammation of the colonic mucosa, scattered and focal lymphocyte infiltration in the local muscle layer, and myositis related changes. Pathological diagnosis was secondary CIPO induced by AEL. (3) Postoperative immune rejection, recurrence and treatment. Results of colonoscopy and histopatholo-gical examination at postoperative 8 days showed acute cellular rejection. The cell count of reci-pient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes were 27.00×10 3, 373.00×10 3, 179.00×10 3 and 142.00×10 3 cell/mL, respectively. Anti-immune rejection treatment was performed using tacrolimus, rabbit anti-human thymocyte immunoglobulin, methylprednisolone mycophenolate mofetil, and monoclonal antibodies against basil. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes at postoperative 57 days were 0.72×10 3, 239.59×10 3, 89.28×10 3 and 91.53×10 3 cell/mL, respectively. Results of colonoscopy and histopathological examination at postoperative 79 days showed the recurrence of AEL. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes were 0.32×10 3, 264.92×10 3, 46.95×10 3 and 169.54×10 3 cell/mL, respectively. The tacrolimus and methylprednisolone were used for treatment. Results of colonoscopy and histopathological examina-tion at postoperative 89 days showed AEL recurrence without remission. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lympho-cytes, and CD8 + lymphocytes were 0.28×10 3, 187.00×10 3, 55.52×10 3 and 92.45×10 3 cell/mL, respec-tively. The tacrolimus and methylprednisolone were used for treatment. Results of colonoscopy and histopathological examination at postoperative 92 days showed the intestinal mucosa had returned to a normal state. (4) Postoperative oral feeding time and time to get rid of parenteral nutrition. The recipient began oral feeding at postoperative 28 days and eliminated parenteral nutrition at postoperative 35 days. (5) Follow-up. The recipient was discharged 114 days after surgery and as of the follow-up deadline, the graft function was good. The recipient maintained a low-fat, high sugar, and high protein diet, completely consumed orally, with a body mass index of 22 kg/m 2, and has returned to normal work. Conclusion:MMT can be used for the treatment of CIPO secondary to AEL.

Objective To explore the role of zinc finger protein 36 (ZFP36) in regulating osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) and preosteoblasts. Methods ZFP36 expression was observed in primary mouse BMSCs and mouse preosteoblasts (MC3T3-E1 cells) during induced osteogenic differentiation. Zfp36-deficient cell models were constructed in the two cells using RNA interference technique and the changes in differentiation capacities of the transfected cells into osteoblasts were observed. Transcriptome sequencing was used to investigate the potential mechanisms of ZFP36 for regulating osteoblast differentiation of the two cells. U0126, a ERK/MAPK signal suppressor, was used to verify the regulatory mechanism of Zfp36 in osteogenic differentiation of Zfp36-deficient cells. Results During the 14-day induction of osteogenic differentiation, both mouse BMSCs and MC3T3-E1 cells exhibited increased expression of ZFP36, and its mRNA expression reached the peak level on Day 7 (P<0.0001). The Zfp36-deficient cell models showed reduced intensity of alkaline phosphatase (ALP) staining and alizarin red staining with significantly lowered expressions of the osteogenic marker genes including Alpl, Sp7, Bglap and Ibsp (P<0.01). Transcriptome sequencing verified the reduction of bone mineralization-related gene expressions in Zfp36-deficient cells and indicated the involvement of ERK signaling in the potential regulatory mechanism of Zfp36. Immunoblotting showed that pERK protein expression increased significantly in Zfp36-deficient cells compared with the control cells. In Zfp36-deficient MC3T3-E1 cells, inhibition of activated ERK/MAPK signaling with U0126 resulted in obviously enhanced ALP staining and significantly increased expressions of osteoblast differentiation markers Runx2 and Bglap (P<0.05). Conclusions ZFP36 is involved in the regulation of osteoblast differentiation of mouse BMSCs and preosteoblasts, and ZFP36 deficiency causes inhibition of osteoblast differentiation of the cells by activating the ERK/MAPK signaling pathway.