Serving as cerebral macrophages， microglial cells are meticulously regulated by the microenvironment of the central nervous system.In response to various environmental and cellular stresses， microglial cells are rapidly activated and exhibit either pro-inflammatory or anti-inflammatory phenotypes to maintain brain tissue homeostasis， and during this process， significant changes are observed in glucose metabolism of microglial cells. Aerobic glycolysis is the primary energy source for pro-inflammatory microglial cells， while oxidative phosphorylation is the energy source for anti-inflammatory microglial cells.This article systematically elaborates on glucose metabolism and glucose metabolic reprogramming pathways in microglial cells， as well as their role in central nervous system diseases. In addition， this article also discusses the potential of targeting glucose metabolic reprogramming in microglial cells for the treatment of related diseases.

Objective To identify genetic regulators of ionizing radiation(IR)sensitivity through clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)genome-wide screening.Methods A specialized single guide RNA(sgRNA)library was developed targeting 987 stress-response regulatory genes identified through Kyoto Encyclopedia of Genes and Genomes(KEGG),Reactome and gene ontology(GO)databases,followed by construction of sgRNA plasmids and packaging into a lentiviral library.Using colon adenocarcinoma Caco-2 cells as a model system,the Cas9-stable transgenic line was established via lentiviral transduction and antibiotic selection.Library-transduced cells underwent puromycin selection,followed by γ-irradiation(dose optimized via preliminary experiments).Post-irradiation phenotypic selection was conducted after 14 days,with subsequent next-generation sequencing of surviving cell populations to identify enriched/depleted sgRNAs.Candidate genes were functionally validated through orthogonal assays:cell counting kit-8(CCK-8)proliferation analysis,clonogenic survival assays and flow cytometric quantification of reactive oxygen species(ROS)and apoptotic markers.Results The optimized screening platform identified 281 radiation response genes(165 radiosensitive and 116 radioprotective candidates).Functional validation revealed Bcl2-associated athanogene 3(BAG3)knockdown significantly enhanced radioresistance.Proliferation was increased 1.2-1.5 fold,clonogenic survival improved 1.5-2.0 fold,and ROS was reduced by 13％-25％coupled with 32％-73％apoptosis attenuation compared to control groups.Conclusion The integrated CRISPR/Cas9 screening platform effectively identifies novel radiation response regulators,as evidenced by the discovery of BAG3 as a critical radiosensitivity determinant.This high-throughput functional genomics approach provides a robust methodology for systematically mapping molecular determinants of cellular radiation response,with potential applications in both mechanistic studies and therapeutic target discovery.

Objective To assay the salivary ANG-2 level of oral lichen planus(OLP)patients,and analyze its correlation with in-flammatory activity of OLP.Methods Eighty-nine OLP patients were included,and divided into four subgroups as non-erosive asymp-tomatic(NEA),non-erosive symptomatic(NES),minor-erosive(MIE)and major-erosive(MAE)groups.Fifteen healthy adults were recruited as controls.Whole unstimulated saliva was collected from each participant,and the salivary ANG-2 level was measured by chemiluminescence immunoassays(CLIA)for analysis.Normal oral mucosal tissue,non-erosive and erosive OLP tissues were collected to detect and analyze the expression of ANG-2 positive blood vessels by immunohistochemistry(IHC).Results The base-lines of age and gender between OLP and control groups showed no significant difference.Compared to controls,the salivary ANG-2 levels of OLP group,non-erosive and erosive OLP subgroups were significantly higher(P＜0.05),in which erosive OLP group was higher than non-erosive OLP group(P=0.022);NES subgroup was slightly higher than NEA(P=0.048),and there was no statistical significance between MIA and MEA subgroups(P=0.067).Spearman correlation analysis showed a positive correlation between sali-vary ANG-2 level and inflammation activity in OLP patients(r=0.314,P=0.003).The expression of ANG-2 in non-erosive OLP mu-cosal tissues slightly increased than normal oral mucosal tissue(P＞0.05),but there was no significant difference.The expression of ANG-2 in erosive OLP mucosal tissues significantly increased than normal oral mucosal tissue(P＜0.001)and non-erosive OLP group(P＜0.001).Conclusion There is a certain correlation between sali-vary ANG-2 level and inflammatory activity of OLP,indicating that salivary ANG-2 level is probable to be one of the inflammatory activity indicators to monitor the state-variation of OLP as a clinical non-inva-sive method.

OBJECTIVE To establish the quality standards of medicinal materials in light of related methods in the general principles of part four of Chinese Pharmacopoeia(2020 Edition), and to conduct systematic research on the Tibetan medicine "Yajima"(Chrysosplenium axillare). METHODS The powder characteristics of medicinal materials were described by microscopic identification method. Silica gel GF254 thin-layer plate was employed to establish a TLC identification method with 5-O-demethylapulein and oxyayanin A as reference substances. Loss on drying, total ash, acid-insoluble ash and ethanol-soluble extractives of 10 batches of Chrysosplenium axillare were determined according to the general principles of part four of Chinese Pharmacopoeia(2020 Edition). HPLC was used to establish the characteristic chromatogram of Chrysosplenium axillare, and the content determination method was established with chrysosplenoside I(CI) and chrysosplenoside A(CA) as the quality control index components of Chrysosplenium axillare. RESULTS The water content, total ash, acid-insoluble ash, ethanol-soluble extractive and the content of CI and CA of all samples varied in the ranges of 9.17%−12.52%, 14.11%−16.74%, 1.50%−4.72%, 32.77%−40.30%, 0.30%−0.99% and 0.28%−0.88%, respectively. CONCLUSION The identification and content determination methods of Yajima(Chrysosplenium axillare) are established for the first time. The methods are easy to operate and exclusive, which is of great significance to accurately evaluate the internal quality of medicinal materials and ensure the quality of drug used.

Objective To observe the influence of different energy windows of the medium-energy general-purpose(MEGP)collimator on image quality,so as to optimize the energy window of yttrium-90(90Y)bremsstrahlung SPECT imaging.Methods 90Y bremsstrahlung spectrum was acquired,and the sensitivity,percentage of the source counts in useful field of view(S/FOV%)and signal-to-background ratio(S/B)of 90Y bremsstrahlung SPECT imaging at MEGP under different energy windows were compared.Results The energy spectrum of 90Y bremsstrahlung was a continuous curve,with the peak of 76.2 keV with MEGP collimator.The images obtained with MEGP collimator were clear,and no significant differences of S/FOV%nor S/B was found between 10%and 20%window width groups(both P＞0.05),but the sensitivities of the latter was higher than the former(P＜0.05).The sensitivity of 70-90 keV images was relatively high,while the S/FOV%and S/B had decreased.The S/FOV%and S/B of images ranging from 40-60 keV were high,but the sensitivity was low.Images acquired with 100 keV±20%showed fairly high sensitivity,S/FOV%and S/B,which was 69.73%,0.62 and 1.64,respectively.Conclusion When performing 90Y bremsstrahlung SPECT with MEGP collimator,the image quality at 20%window width was better than at 10%window width,and 100 keV±20%showed fairly high sensitivity and not significantly decreased S/FOV%and S/B.

Objective:To study the distribution and changes of dopamine transporter (DAT) in guinea pig eyes under different light patterns.Methods:Thirty-six 3-week-old white ordinary-grade guinea pigs were randomly selected and divided into groups of 10 000 lx, 5 000 lx, and 500 lx, with 12 guinea pigs in each group exposed to strong light, medium strong light, and normal light, respectively.Each group was randomly divided into 3 subgroups, with 4 guinea pigs in each subgroup.The 3 subgroups of 500 lx group received light exposure for 5, 20, and 40 minutes, respectively.The 3 subgroups of 5 000 lx group received light exposure for 2, 4, and 40 minutes, respectively.The 3 subgroups of 10 000 lx group received light exposure for 2, 5, and 20 minutes, respectively.After light treatment, each group of guinea pigs was injected with 99mTc-TRODAT-1 for SPECT DAT imaging, and image data were collected by Micro-SPECT.The region of interest (ROI) of guinea pig retinas was analyzed using Micro-CT software.The counts of ROI were expressed as Sum, which reflected the relative distribution or density of DAT.The DAT density between experimental and control eyes of guinea pigs after light exposure, the differences in DAT density between guinea pig eyes under different light intensities, the differences in DAT density between guinea pig eyes after different light durations, and the cumulative and interactive effects of light intensity and light duration on DAT aggregation in guinea pigs were compared.Another 3 guinea pigs were selected, and after light exposure, the 3 guinea pigs' eyes underwent continuous image acquisition for 6 hours at 20-minute intervals, and 18 images per guinea pig were acquired to analyze the trend of DAT density in guinea pig eyes over time.This study was approved by the Ethics Committee of Shanghai General Hospital (No.2020SQ196). Results:The DAT density (Sum value) of experimental eyes at 500, 5 000, and 10 000 lx were 5 598.97±3 159.38, 8 636.78±2 503.16, and 7 407.39±2 053.41, respectively, significantly higher than 4 388.89±2 902.90, 5 981.92±3 057.44, and 5 091.32±2 039.36 of control eyes ( t=5.31, 4.69, 11.80; all at P<0.001). At 500 lx, there was a statistically significant difference in DAT density between the experimental and control eyes of guinea pigs at different light exposure durations ( F=14.01, P<0.01), while no significant difference was found at other light intensities at different light exposure durations (both at P>0.05). When the light exposure time was 5 minutes, the difference in DAT density between the experimental and control eyes of guinea pigs was significantly greater in the 10 000 lx group than in the 500 lx group ( t=-13.22, P<0.001). There was no statistically significant difference between different groups at other light exposure durations (all at P>0.05). No cumulative or interactive effects of light intensity and light duration were found on the differences in DAT density (all at P>0.05). Continuous scanning after illumination showed that DAT density in guinea pig retinas first increased to a peak over time and then gradually returned to normal values. Conclusions:Light, even under moderate or normal light levels, can cause an increase in the secretion of DAT in the retina and stimulate the production of DAT.Light intensity and duration have no cumulative or interactive effects on the distribution and density of retinal DAT.

Currently,human health due to corona virus disease 2019(COVID-19)pandemic has been seriously threatened.The coronavirus severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike(S)protein plays a crucial role in virus transmission and several S-based therapeutic approaches have been approved for the treatment of COVID-19.However,the efficacy is compromised by the SARS-CoV-2 evolvement and mutation.Here we report the SARS-CoV-2 S protein receptor-binding domain(RBD)inhibitor licorice-saponin A3(A3)could widely inhibit RBD of SARS-CoV-2 variants,including Beta,Delta,and Omicron BA.1,XBB and BQ1.1.Furthermore,A3 could potently inhibit SARS-CoV-2 Omicron virus in Vero E6 cells,with EC50 of 1.016 pM.The mechanism was related to binding with Y453 of RBD deter-mined by hydrogen-deuterium exchange mass spectrometry(HDX-MS)analysis combined with quan-tum mechanics/molecular mechanics(QM/MM)simulations.Interestingly,phosphoproteomics analysis and multi fluorescent immunohistochemistry(mIHC)respectively indicated that A3 also inhibits host inflammation by directly modulating the JNK and p38 mitogen-activated protein kinase(MAPK)path-ways and rebalancing the corresponding immune dysregulation.This work supports A3 as a promising broad-spectrum small molecule drug candidate for COVID-19.

Objective:To explore the clinical features and genetic basis for a fetus featuring Rhizomelic skeletal dysplasia.Methods:A fetus diagnosed at the Reproductive and Genetic Center of Suzhou Municipal Hospital in November 2020 was selected as the study subject. Whole exome sequencing (WES) was carried out for the fetus and its parents. Candidate variants were verified by Sanger sequencing. Peripheral blood smears of both parents were also examined.Results:The fetus was found to have a small chest and short limbs, which had suggested skeletal dysplasia. Genetic testing revealed that the fetus has harbored compound heterozygous variants of the LBR gene, including a paternally derived c. 1687+ 1G>A and a maternally derived c. 1757G>A (p.Arg586His). The blood smear of the father showed Pelger-Huёt anomaly with hyposegmentation of neutrophil nuclei, while the neutrophils of the mother appeared to be normal. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG) and the Association for Molecular Pathology (AMP), the c. 1757G>A (p.Arg586His) variant was classified as likely pathogenic (PM3_Strong+ PM2_Supporting+ PP3), and so was the c. 1687+ 1G>A variant (PVS1-Moderate+ PM3+ PM2-Supporting+ PP4). Conclusion:The compound heterozygous variants of the LBR gene probably underlay the pathogenesis of skeletal dysplasia in this fetus.

Objective:To explore the clinical characteristics and genetic basis for a fetus with Joubert syndrome.Methods:A pregnant woman who had visited Suzhou Municipal Hospital on February 26, 2021 was selected as the study subject. The fetus and her parents were subjected to whole exome sequencing (WES), and candidate variants were verified by Sanger sequencing. cDNA analysis of her father and RNA sequencing of her sister were also carried out.Results:The fetus was found to harbor compound heterozygous variants of the TCTN1 gene, namely c. 624G>A and c. 96dupA (p.Glu33Argfs*49), which were inherited from her father and mother, respectively. Her sister also carried the paternal c. 624G>A variant, and mRNA transcripts with the c. 624G>A variant of the TCTN1 gene were not detected by cDNA analysis of her father and RNA sequencing of her sister. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c. 624G>A and c. 96dupA variants were both classified as likely pathogenic (PVS1+ PM2_Supporting). Conclusion:The compound heterozygous variants of the TCTN1 gene probably underlay the pathogenesis in this fetus. Above finding has also expanded the mutational spectrum of the TCTN1 gene.

Objective To analyze the diagnostic value of systemic immune inflammation index(SII)and systemic inflammation response index(SIRI)in rheumatoid arthritis(RA).Methods A total of 470 RA pa-tients diagnosed in the hospital from January to December 2023 were selected.According to the results of anti-cyclic citrullinated peptide(anti-CCP)antibody and rheumatoid factor(RF),RA was divided into four groups:anti-CCP antibody and RF negative group(52 cases),anti-CCP antibody positive RF negative group(70 cases),anti-CCP antibody negative RF positive group(18 cases),anti-CCP antibody and RF positive group(330 cases),and 300 healthy subjects were selected as control group.After calculating SII and SIRI in each group,Kruskal-Wallis was used to examine the difference between SII and SIRI in the anti-CCP antibody and RF negative group,anti-CCP antibody positive RF negative group,anti-CCP antibody negative RF positive group,anti-CCP antibody and RF positive group of RA patients and the control group,and the receiver operat-ing characteristic(ROC)curve was drawn to evaluate the diagnostic efficacy of SII and SIRI in the anti-CCP antibody and RF negative group,anti-CCP antibody positive RF negative group,anti-CCP antibody negative RF positive group,anti-CCP antibody and RF positive group of RA patients.Results SII and SIRI in four groups of RA patients were significantly higher than those in control group(P＜0.05).The area under the curve(AUC)of SII in evaluating the four groups of RA patients were 0.689,0.724,0.709,and 0.774,respec-tively.SIRI had an AUC of 0.679,0.729,0.679,and 0.772 in the evaluation of the four groups of RA pa-tients,respectively.Conclusion SII and SIRI are both elevated in patients with RA,and have certain predictive value for RA.