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.

Thanks to the application of gene modification technology, the development of new immunosuppressant and the shift towards using pigs as the main organ donors, xenotransplantation has embarked on a new round of "ice-breaking journey". Xenotransplantation demonstrates strong positive value in addressing the imbalance between organ supply and demand, extending the lives of patients with organ failure and enhancing human physical functions. However, xenotransplantation also raises a series of ethical risks, such as safety concerns over health damage and virus transmission, the hollowing out of the principle of informed consent, the impairment of individual identity and independence, and the distribution fairness of access and capability for trials. To address these issues, a mechanism combining prior general consent and specific consent during the process should be established, along with a dynamic and traceable consent procedure to safeguard patient autonomy. A "waterfall-style" regulatory system and a mechanism for aggregating cutting-edge technologies should be created to enhance the safety of xenotransplantation. Distribution and governance rules should be formulated according to different stages and goals of xenotransplantation exploration to improve fairness. A typological approach should be used to determine which groups of people can undergo xenotransplantation under current medical conditions to achieve the minimization of harm.

Objective: To investigate the effects of RING 1 and YY1 binding protein (RYBP), a member of the polycomb group (PcG), on bladder cancer cell proliferation and invasion, as well as the underlying mechanisms involved, so as to provide reference for the prevention and treatment of bladder cancer. Methods: Overexpressing and knocking down of RYBP were achieved in bladder cancer cell lines (T24, UM-UC-3, and 5637) via plasmids or siRNAs.Cell proliferation was assessed via thiazolyl blue (MTT) and colony formation assays, whereas migration was evaluated via scratch and Transwell assays.Changes in epithelial-mesenchymal transition (EMT) markers and other related proteins were examined with Western blotting. Results: RYBP overexpression significantly inhibited bladder cancer cell proliferation, invasion, and migration, whereas RYBP knockdown promoted these behaviors.Western blotting results revealed that RYBP overexpression downregulated the expressions of EMT markers N-cadherin, Vimentin, and Slug, but upregulated the expression of E-cadherin.Conversely, RYBP knockdown upregulated the expressions of N-cadherin, Vimentin, and Slug, while reducing the expression of E-cadherin. Conclusion: RYBP appears to inhibit the proliferation and migration of bladder cancer cells via the EMT pathway, indicating its potential application in bladder cancer therapies.

Objective: To investigate the spatio-temporal clustering characteristics of influenza in Ningxia Hui Autonomous Region from 2014 to 2023, so as to provide the basis for strengthening influenza prevention and control. Methods: Data pertaining to influenza cases reported in Ningxia Hui Autonomous Region from 2014 to 2023 were retrieved from the Infectious Disease Surveillance System of the Chinese Disease Prevention and Control Information System, including age, sex, current residence, onset date, and reporting date. The seasonal incidence of influenza was analyzed using seasonal index. The spatio-temporal clustering characteristics of influenza were identified using spatial autocorrelation analysis and spatio-temporal scan analysis. Results: A total of 20 377 influenza cases were reported in Ningxia Hui Autonomous Region from 2014 to 2023, with a male-to-female ratio of 1.15∶1. The majority were children under 15 years, with 10 950 cases accounting for 53.74%. Influenza was highly prevalent in January, February, March, and December, with seasonal indices of 219.06%, 111.00%, 246.65%, and 366.24%, respectively. The average annual reported incidence was 29.55/100 000, among which Pengyang County, Jinfeng District, Dawukou District, Xiji County, and Litong District had higher average annual reported incidence, at 63.99/100 000, 55.71/100 000, 55.70/100 000, 49.49/100 000, and 49.04/100 000, respectively. Spatial autocorrelation analysis showed that in 2023, there was spatial clustering of influenza cases in Ningxia Hui Autonomous Region (Moran's I=0.333, P<0.05), with a high-high cluster in Jingyuan County, while in other years, the distribution of influenza cases was random (all P>0.05). Spatio-temporal scan analysis showed that from 2014 to 2023, there were four space-time clusters in Ningxia Hui Autonomous Region, including one type Ⅰ cluster in Hongsibao District of Wuzhong City, with the clustering period from January 20 to 26, 2014; and three type Ⅱ clusters, mainly in January, February, March and December, covering one area in Shizuishan City, five areas in Guyuan City, one area in Zhongwei City, three areas in Wuzhong City, and four areas in Yinchuan City. Conclusions From 2014 to 2023, children under 15 years were the primary population affected by influenza in Ningxia Hui Autonomous Region, with distinct spatio-temporal distribution characteristics. The peak incidence occurred during the winter and spring seasons, and the main clustering areas were in the southern regions.

Objective To investigate the biological function of long-stranded non-coding RNA colorectal tumor differentially expressed gene(lncRNA CRNDE)in the malignant progression of papillary thyroid carci-noma(PTC)and its potential regulatory mechanisms.Methods Cancer and paracancerous tissue specimens from totally 30 PTC patients who underwent thyroidectomy at the Affiliated People's Hospital of Jiangsu U-niversity from July 2019 to January 2021 were collected,and human thyroid cancer cell lines(TPC-1 and BC-PAP)and normal human thyroid cells(Nthy-ori 3-1)were additionally selected as the target cells.Real time quantitative polymerase chain reaction(qPCR)was performed to detect the expression of lncRNA CRNDE and microRNA(miR)-451 in PTC cancer,paracancerous tissues,and cell lines.TPC-1 and BCPAP cells were randomly divided into control(Ctrl)group,lncRNA CRNDE overexpression(lncRNA CRNDE-OE)group and lncRNA CRNDE knockdown(lncRNA CRNDE-siRNA)group.CCK-8 method was used to detect cell proliferation activity.Transwell method was used to detect cell invasion and migration.Flow cytometry was used to detect cell apoptosis.Western blot was used to detect N-cadherin,vimentin,and E-cadherin protein ex-pression.The luciferase reporter assay was used to analyze the targeting relationship between lncRNA CRNDE and miR-451 gene.Results lncRNA CRNDE expression was increased and miR-451 expression was decreased in cancer tissues compared with paracancerous tissues(P＜0.001);lncRNA CRNDE expression was up-regulated and miR-451 expression was down-regulated in TPC-1 and BCPCP cells compared with Nthy-ori 3-1 cells(P＜0.001).Compared with the Ctrl group,BCPAP and TPC-1 cells in the lncRNA CRNDE-OE group showed increased survival,invasion and migration rates,decreased apoptosis,decreased E-cadherin pro-tein expression,elevated N-cadherin and Vimentin protein expression,and decreased miR-451 expression(P＜0.01),whereas in the lncRNA CRNDE-siRNA group,cell survival,invasion rate and migration rate were de-creased,apoptosis rate was elevated,E-cadherin protein expression was increased,and N-cadherin and Vimen-tin protein expression was decreased(P＜0.01).Dual luciferase reporter assay showed that lncRNA CRNDE had obvious targeting relationship with miR-451.Conclusion lncRNA CRNDE is highly expressed in PTC cancer tissues,which may promote PTC cell proliferation,migration,invasion,epithelial mesenchymal transi-tion,and inhibit cell apoptosis by targeting miR-451 expression inhibition.

A number of drugs for the treatment of type 2 diabetes mellitus(T2DM)are currently under clinical investigation,including the sodium-dependent glucose transporters 2(SGLT2)inhibitor rongliflozin,the SGLT1/2 inhibitor LIK066,the di-peptidyl peptidase-4(DPP-4)inhibitor DBPR108,the glucagon-likepeptide-1 receptor(GLP-1R)ago-nist CJC-1134-PC,the G-protein-coupled receptor 40(GRP40)agonist SCO-267 and the Glucokinase(GK)agonist PB201.This article briefly reviews the clinical research progress of drugs targeting the above targets in the field of T2DM treatment,in or-der to provide reference for the treatment of T2DM patients.

Objective:To explore the neuroprotective effects and mechanisms of memantine on sepsis-associated encephalopathy (SAE) model mice.Methods:Totally 90 male C57BL/6J mice aged 8-12 weeks were randomly divided into 3 groups (with 30 mice in each group) : sham group, model group and memantine group. The SAE mouse model was established by cecum ligation and puncture while mouse in sham group received open and closed abdomen only. The mice in the memantine group were irrigation with memantine (15 mg · kg -1· d -1) 3 hours before surgery and 7 consecutive days after modeling. The mice in the model group and sham group were irrigation with an equal volume of 0.9% sodium chloride solution. The 7-day survival rate was observed, neurobehavioral and cognitive function scores of each group of mice after modeling were assessed.Blood-brain barrier permeability was measured by detecting the content of Evans blue. Immunofluorescence staining was used to detect the expression of astrocytes. Enzyme linked immunosorbent assay was conducted to detect cellular inflammatory factors and the glutamic acid content detection kit was used to detect the expression of glutamic acid. All data were analyzed by Graphpad Prism 8.3.0 software, survival rate was analyzed using Kaplan-Meier survival curve.Multigroup comparisons were conducted by one-way ANOVA or Kruskal-Wallis test. Results:(1) There was a statistically significant difference in the 7-day survival rate among the three groups of mice after modeling ( F=24.11, P<0.01), and the 7-day survival rate of the memantine group was higher than that of the model group (57% (17/30), 27% (8/30), P<0.01). (2)The behavioral results showed that after 7 days of modeling, there were statistically significant differences in the total distance of the open field test, central area stay time, four corner area stay time, neurobehavioral scores, pole climbing test, and preference index for new object recognition test among the three groups of mice ( F/ χ2=17.67, 17.30, 9.39, 14.06, 10.36, 14.81, all P<0.05).The neurobehavioral score, pole climbing test score, preference index for new object recognition test, total distance of open field test, and central area stay time of the model group were all lower than those of the sham group (all P<0.05), while four corner area stay time of the model group was higher than that of the sham group ( P<0.05).The total distance of open field test (1 564.07(1 363.24, 1 988.19) cm, 913.91 (574.32, 1 096.23) cm), central area stay time (5.21 (4.91, 8.76) s, 1.09 (0.25, 1.64) s), neurobehavioral scores (9.75±0.50, 8.25±0.50), pole climbing test scores (5.67±0.52, 4.56±0.53), and preference index for new object recognition test (56.50±10.59, 26.84±2.91) of the memantine group were all higher than those of the model group (all P<0.05). The four corner area stay time was lower than that of the model group ((480.30±50.64) s, (529.80±36.20) s, P<0.05).(3)The comparison of molecular indicators showed that there were statistically significant differences in the content of Evans blue in the brain, the number of astrocytes in the hippocampus and cerebral cortex, pro-inflammatory cytokines (TNF-α、IL-1β、IL-6), anti-inflammatory cytokines (IL-10), and glutamic acid among the three groups of mice ( F/ χ2=8.84, 6.43, 28.46, 23.63, 12.23, 16.04, 69.22, 6.65, all P<0.05).The content of Evans blue, the number of astrocytes in the hippocampus and cerebral cortex, the expression of TNF-α、IL-1β、IL-6, and glutamate in the model group were all lower than those in the sham group(all P<0.05). The levels of IL-10 in the model group was lower than that in the sham group ( P<0.05).The content of Evans blue ((5.67±1.38)μg/g, (11.08±2.79)μg/g), the number of astrocytes in the hippocampus (16.50 (13.75, 22.25)/μm 2), 80.00 (73.50, 83.50)/μm 2) and the cerebral cortex (40.00 (29.00, 48.00)/μm 2, 81.50 (72.25, 89.00)/μm 2) in the memantine group were lower than those in the model group (all P<0.05).The pro-inflammatory factor TNF-α, IL-1β, IL-6 and glutamic acid expression in the memantine group were lower than those in the model group (all P<0.05), and the anti-inflammatory cytokine IL-10 was higher than that in the model group ( P<0.05). Conclusions:Memantine can improve the neurobehaviors and cognitive functions of SAE mice through improving the integrity of the damaged blood-brain barrier, alleviating inflammation in the brain, as well as reducing glutamate levels in the brain.

Improving liver regeneration (LR) remains a medical issue, and there is currently a lack of safe and effective drugs for LR. Rhizoma Dioscoreae (SanYak, SY) is a traditional Chinese medicine. However, the underlying action mechanism of SY treatment for LR is yet to be fully elucidated. To explore the mechanism by which SY affects LR, we have conducted a series of methods for network pharmacological analysis, molecular docking, and in vivo experimental validation in mice. Overall, 9 compounds and 30 predicted target genes of SY were found to be associated with the therapeutic effects of LR. Compared with the model group, hematoxylin and eosin staining revealed that the mice with preoperative drug intervention possessed fewer postoperative hepatocyte bubbles and relatively regular morphology. Furthermore, the serum alanine transaminase and aspartate aminotransferase levels were reduced, immunohistochemistry revealed elevated proliferating cell nuclear antigen positivity rate, and Western blotting demonstrated that the phospho-protein kinase B (AKT)/AKT ratio was downregulated and that vascular endothelial growth factor A (VEGFA) expression levels were upregulated. This study explored dioscin, the main active ingredient of SY, and its potential therapeutic effects on LR. It repairs damaged liver following surgery and promotes liver cell proliferation. The action mechanism comprises reducing AKT phosphorylation levels and upregulating VEGFA expression levels. Thus, this study provides a new direction for further research on the mechanism of SY promoting LR.

Objective:To explore the impact of early and late antibody-mediated rejection (AMR) on treatment options and allograft outcomes after kidney transplantation (KT).Methods:From January 2013 to December 2022, the study retrospectively enrolled 141 KT allograft recipients receiving allograft biopsy and diagnosed as AMR according to the Banff 2019 criteria. Recipients with a diagnosis of AMR within 30 days post-KT were classified into early AMR group (n=19) while the remainders assigned as late AMR group (n=122). The outcome endpoints included recipient survival rate, death-censored graft survival rate, follow-up estimated glomerular filtration rate (eGFR) and immunodominant donor-specific antibody (DSA) intensity. Wilcoxon's test was utilized for assessing the differences in eGFR and DSA intensity while Kaplan-Meier curve and Log-rank test were employed for evaluating graft survival impact. Treatment regimens for AMR were collected and categorized.Results:The median follow-up duration was 2.6(1.2, 5.2) year. No graft failure was noted in early AMR group while 44 recipients in late AMR group experienced graft failure, with 34 cases (77.2%) due to AMR progression. The 5-year death-censored graft survival rate was significantly better in early AMR group than that in late AMR group [100% vs 60.1%(50.5%, 71.6%), P=0.002]. The one-year change in eGFR for early AMR group was significantly superior to that of late AMR group [19.3(-2.6, 38.1) vs -3.3(-14.0, 5.4), P=0.001]. One-year mean fluorescent intensity (MFI) of early AMR group was 1 158(401.5, 3 126.5). It was significantly lower than that when diagnosed with early AMR [3 120.5(2 392.8, 9 340.0)] and one-year MFI of late AMR group [8 094(2 251.5, 13 560.5)] ( P=0.005, P<0.001). Early AMR group primarily received standard treatment (3/19, 15.8%) and regimens centered on rituximab and/or bortezomib (7/19, 43.8%). Late AMR group mainly received standard (16/122, 13.1%) or intensified regimens (9/122, 7.4%) and regimens focused upon rituximab and/or bortezomib (32/122, 26.2%) and MP monotherapy (21/122, 17.2%). Conclusion:The outcome for early AMR is significantly better than that for late AMR. For early AMR, early and robust immunosuppression is recommended. For late AMR, early detection and timely treatment are crucial and individualized strategies should be implemented.

Objective To explore the effective target and active ingredient prediction of premature ovarian insufficiency(POI)treated by Tiaojing decoction by ferroptosis pathway by network pharmacokogy and bioniformatics.Methods The active ingredients and target genes of Tiaojing decoction were obtained through traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)and bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine databases,disease targets for POI were obtained by using GeneCards,therapeutic target database and Drugbank,and intersection targets were obtained by Venn.Core target gene network of Tiaojing decoction in treatment of POI were constructed through the ferroptosis pathway by ferroptosis database,Cytoscape and STRING software.Then,the DAVID database was used to perform gene ontology and biological pathway enrichment analysis on the predicted targets related to ferroptosis of POI treated with Tiaojing decoction.Subsequently,differential genes in biology related to POI were obtained through the gene expression omnibus(GEO)database for further extraction of key targets,and the binding affinity of active small molecule drug components were verified through molecular docking.Results A total of 154 active ingredients,101 targets related to POI and 23 genes related to ferroptosis were selected from 13 kinds of traditional Chinese medicines in Tiaojing decoction.The enrichment analysis showed that the main involved pathways were phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B(PI3K/Akt)signaling pathway and hypoxia inducible factor-1 signaling pathway.Through GEO database screening,the key targets of Tiaojing decoction in intervening POI through ferroptosis pathway were glycogen synthase kinase-3 β(GSK-3 β),caveolin-1(CAV1),mammalian target of rapamycin(mTOR)and protein kinase C alpha gene(PRKCA),all expression of which reduced in POI.The results of molecular docking showed that CAV1-quercetin and GSK-3β-luteolin had stable binding ability.Conclusion The network pharmacology results suggest that the ferroptosis pathway may be an important mechanism of Tiaojing decoction in treatment of POI,GSK-3β,CAV1,mTOR,PRKCA and other targets,as well as PI3K/Akt signaling pathway,may play important roles in them.