1.Synergistic diagnostic potential of novel biomarkers SYWC and ADA for tuberculous pleural effusion
Chunhong FAN ; Wenlong HU ; Limei ZHU ; Ruifu YANG ; Daxia CAI ; Shaohua LU ; Xingdong CAI
The Journal of Practical Medicine 2025;41(15):2406-2411
Objective To evaluate the respective or synergistic value of cytoplasmic tryptophan-tRNA ligase(WARS1/SYWC)and adenosine deaminase(ADA)in diagnosing tuberculous pleural effusion(TPE).Methods A retrospective analysis was conducted on 120 patients with pleural effusion(64 cases of TPE,56 cases of non-TPE)admitted to the First Affiliated Hospital of Jinan University and its affiliated Shunde Hospital from January 2020 to December 2024.Pleural fluid SYWC levels were identified using enzyme-linked immunosorbent assay(ELISA).Univariate and multivariate logistic regression analyses were performed to identify diagnostic predictors,while receiver operating characteristic(ROC)curves were plotted to assess the diagnostic perfor-mance of individual and combined biomarkers.Results Compared to the non-TPE group,TPE group exhibited significantly younger age,lower pleural CEA,less serum CEA,and lower neutrophil-to-lymphocyte ratio(NLR),but significantly higher levels of pleural ADA,total protein,SYWC,and serum CRP(all P<0.05).Univariate analysis identified age,pleural CEA,carbohydrate antigen 199,ADA,SYWC,serum CEA,and NLR as potential predictors.Multivariate analysis confirmed pleural ADA(OR=1.064,95%CI:1.017~1.228)and SYWC(OR=6.695,95%CI:2.794~16.04)as independent diagnostic factors.At optimal cutoffs,SYWC(16.94 μg/L)demonstrated a sensitivity of 71.80%and specificity of 98.21%,while ADA(36.5 U/L)showed a sensitivity of 93.75%and a specificity of 89.29%.Combined detection increased the sensitivity to 95.56%,the specificity to 98.0%,and the accuracy to 97.87%.ROC analysis revealed an AUC of 0.973(95%CI:0.943~1.000)for the combination,outperforming ADA(0.897)and SYWC(0.938)alone.Conclusion The combi-nation of SYWC and ADA notably enhances diagnostic efficacy for TPE,providing high sensitivity and specificity as a reliable tool for clinical differentiation.
2.Assessing the Causal Association of Circulating Amino Acids With Myasthenia Gravis:A Bi-Directional Mendelian Randomization Study
Hu ZANG ; Xiaoyu JI ; Chang ZHU ; Wenlong YAO ; Li WAN ; Tongtong LIU
Herald of Medicine 2025;44(3):440-445
Objective To investigate the bidirectional causal relationship between circulating amino acid levels and the risk of myasthenia gravis(MG)using Mendelian randomization(MR).Methods A two-sample Mendelian randomization a-nalysis was conducted using publicly available genome-wide association study(GWAS)genetic data,with validation from GWAS data from different sources to assess the robustness of the results.Five models were used for the two-sample bidirectional MR anal-ysis,and odds ratios(OR)were calculated to evaluate the causal relationship between the levels of nine circulating amino acids and MG risk.Sensitivity analyses,heterogeneity tests,and pleiotropy tests were performed to assess the robustness of the results.The causal effect estimated by the inverse variance weighted(IVW)method was the primary result,and the IVW-estimated causal effects were further validated using data from different GWAS sources to assess robustness.Results Genetically predicted high-er circulating glutamine levels were significantly associated with a lower risk of MG[OR(95%CI)=0.696(0.524,0.926),P=0.012 7,IVW model].Validation analyses using GWAS data from various sources also demonstrated a significant negative associa-tion between genetically predicted higher circulating glutamine levels and MG risk[OR(95%CI)=0.321(0.178,0.581),P=1.67x10-1,IVW model].Moreover,genetically predicted higher MG risk was associated with lower levels of circulating glutamine and alanine(β=-0.178±0.009,P=0.049;β=-0.013±0.007,P=0.048,IVW model,respectively).Conclusion Genetic evidence reveals a potential bidirectional causal relationship between circulating amino acid levels and MG risk.Further studies are required to elucidate the mechanisms underlying this relationship.
3.The general pattern analysis of cirrhosis treatment based Ren-yin year's cases
Yirong HU ; Yixuan LIU ; Wenlong ZHENG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2025;32(4):448-453
Objective A data mining method was used to analyse and summarize Professor Zheng Wenlong's ideas and prescription and medication rules for the treatment of cirrhosis,taking the case of cirrhosis of the liver in Ren-yin year(2022)as an example.Methods A collection of traditional Chinese medicine prescriptions for liver cirrhosis patients who received treatment at the clinic of chief traditional Chinese medicine physician Zheng Wenlong at Hangzhou Traditional Chinese Medicine Hospital of Zhejiang Chinese Medical University from January 20,2022 to January 19,2023,and who met the inclusion and exclusion criteria.An excel database was created to standardize the data,followed by frequency statistical analyzes of prescriptions,medications and the characteristics of herbal remedies.Association rule analysis was carried out using SPSS Modeler 18,which facilitated the creation of complex network diagrams,subsequently enhanced through visualization in software Cytoscape.Clustering analysis was executed utilizing SPSS 25.0.Results A total of 176 prescriptions were selected,and the top 3 most frequently used prescriptions were Lingzhu decoction,Shengming decoction and Beihua decoction,with the frequency of 10.13%(23/227),10.13%(23/227)and 9.69%(22/227).A diverse array of 155 herbal medicines were employed,and the top 3 most frequently used drugs were Glycyrrhiza,Poria and Zingiberis Rhizoma Recens,with the frequency of 4.75%(127/2 676),4.26%(114/2 676)and 3.21%(86/2 676).The predominant medicinal properties of these herbs were classified as either cold or warm,while the primary tastes identified were bitter,sweet and spicy,with channel tropism were liver,lung,stomach and spleen.According to the descending order of improvement percentage,the results showed that the strongly correlated pairs of herbs were Chaenomelis Fructus-Aconiti Lateralis Radix Praeparata and Citri Reticulatae Pericarpium Viride-Pericarpium Citri Reticulatae,with improvement degrees of 2.74%and 2.55%respectively;the strongly correlated 3-herb combinations(Jiao Yao)included Aconiti Lateralis Radix Praeparata-Poria-Chaenomelis Fructus and Aconiti Lateralis Radix Praeparata-Rhizoma Atractylodis Macrocephalae-Chaenomelis Fructus,with improvement degrees of 3.24%and 3.23%respectively.The clustering analysis categorized the high-frequency herbs into 3 distinct groups:C1:Citri Reticulatae Pericarpium Viride,Pericarpium Citri Reticulatae,Jujubae Fructus,Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine,and Zingiberis Rhizoma Recens;C2:Rhizoma Atractylodis Macrocephalae,Aconiti Lateralis Radix Praeparata,Artemisiae Scopariae Herba,and Paeoniae Radix Alba;C3:Schisandra Chinensis,Rhizoma Alismatis,Radix Glycyrrhizae,Poria,Zingiberis Rhizoma and Ramulus Cinnamomi.These correspond to the formulas of Erchen decoction,Yinchenshufu decoction and Wuling powder.Conclusions The underlying pathogenesis of cirrhosis is linked to dysfunctions of the Jueyin and Yangming meridians,as well as disturbances in the Shaoyang axis.The primary therapeutic approach of Professor Zheng Wenlong involves a comprehensive evaluate all of the patient's clinical presentation to identify the shared pathogenesis underlying various symptoms,integrating the patient's personal constitution with the annual cycle of the Five Phases(Wuyun)and Six Qi(Liuqi)as described in traditional Chinese medicine,and formulate prescriptions based on the interactions of the Five Phases(Wuxing)and guided by the yin-yang theory of"Opening-Closing-Pivot"(Kai-He-Shu).The focus of prescriptions is rehabilitation and the final goal is reach the state of"Tian Ren He Yi".
4.Changes in brain activity in patients with post-traumatic stress disorder two months after the traumatic event
Luodong YANG ; Haohao LI ; Yao MENG ; Min HU ; Wenlong XING ; Liang JIANG ; Guiqing ZHANG
Chinese Mental Health Journal 2025;39(4):301-307
Objective:To explore changes in brain activity in patients with post-traumatic stress disorder(PTSD).Methods:A total of 40 participants involved in car accidents were included,and functional magnetic reso-nance imaging(fMRI)scans were collected within one week.Anxiety,depression,and personality assessments were conducted with the Hamilton Anxiety Scale(HAMA),Hamilton Depression Scale(HAMD),and Eysenck Person-ality Scale for Adult(EPQ).After two months,a second fMRI scan was conducted,and a PTSD diagnosis was made.Participants were divided into a trauma-exposed group(n=23)and a PTSD group(n=17)based on wheth-er they developed PTSD.Changes in brain functional activity between the trauma-exposed group and the PTSD group were compared using the percentage of amplitude fluctuation(perAF)method.Results:Compared to the trauma-exposed group,the PTSD group showed a decreased perAF value in the left hippocampus at 1 week,and de-creased perAF values in the right mid-cingulate gyrus and left postcentral gyrus at 2 months(P<0.05).When comparing the PTSD group at different times,the perAF values in the left middle temporal gyrus and left medial su-perior frontal gyrus decreased at 2 months(P<0.05).Correlation analysis revealed that PCL-5 scores were posi-tively correlated with EPQ Psychoticism(r=0.32,P=0.041),HAMA(r=0.35,P<0.05),and HAMD(r=0.34,P<0.05).Regression analysis found that higher scores of EPQ psychoticism(OR=11.79)and HAMA(OR=1.62)were risk factors for post-accident PTSD,while higher scores of EPQ extraversion(OR=0.32)were pro-tective factors.Conclusion:It suggests that patients with post-traumatic stress disorder may show decreased activity in the right middle cingulate cortex,left postcentral gyrus,left middle temporal gyrus,and left medial superior fron-tal gyrus within two months after the traumatic event.
5.An analysis of the seasonal epidemic characteristics of influenza in Kunming City of Yunnan Province from 2010 to 2024
Zexin HU ; Min DAI ; Wenlong LI ; Minghan WANG ; Xiaowei DENG ; Yue DING ; Hongjie YU ; Juan YANG ; Hong LIU
Shanghai Journal of Preventive Medicine 2025;37(8):643-648
ObjectiveTo characterize the seasonal patterns of influenza in Kunming City, Yunnan Province before, during, and after the COVID-19 pandemic, and provide scientific evidence for optimizing influenza prevention and control strategies. MethodsInfluenza-like illness (ILI) and etiological surveillance data for influenza from the 14th week of 2010 to the 13th week of 2024 in Kunming City of Yunnan Province were collected. Harmonic regression models were constructed to analyze the epidemic characteristics and seasonal patterns of influenza before (2010/2011‒2019/2020 influenza seasons), during (2020/2021‒2022/2023 influenza seasons), and after (2023/2024 influenza season) the COVID-19 pandemic. ResultsBefore the COVID-19 pandemic, influenza in Kunming City mainly exhibited an annual cyclic pattern without a significant semi-annual periodicity, peaking from December to February of the next year, with an epidemic duration of 20‒30 weeks. During the pandemic, influenza seasonality shifted, with an increase in semi-annual periodicity and an approximate one month delay in annual peaks. However, after the pandemic, the annual amplitude of influenza increased compared with that before the pandemic, and the epidemic duration extended by about one month. Although the annual peak largely reverted to the pre-pandemic levels, the annual peaks for different influenza subtypes/lineages had not fully recovered. ConclusionInfluenza seasonality in Kunming City underwent substantial alterations following the COVID-19 pandemic and has not yet fully reverted to pre-pandemic levels. Continuous surveillance on different subtypes/lineages of influenza viruses remains essential, and prevention and control strategies should be adjusted and optimized in a timely manner based on current epidemic trends.
6.Expression of m6A-related proteins in mice with PTSD-like behavior improved by sertraline hydrochloride treatment
Jiaying LU ; Luodong YANG ; Min HU ; Wenlong XING ; Haiteng CUI ; Guiqing ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(11):961-968
Objective:To explore the expression changes of N6-methyladenosine (m6A)-related proteins in the hippocampus of mice with post traumatic stress disorder (PTSD)-like behavior and the therapeutic effects of sertraline hydrochloride.Methods:Male C57BL/6J mice aged 4-6 weeks were selected to establish a PTSD model using a single prolonged stress and foot shock stimulation. A total of 24 mice were randomly divided into the control group, model group, and sertraline group using a random number table, with 8 mice in each group. Mice in the sertraline group were intraperitoneally injected with sertraline hydrochloride (15 mg/kg, once daily) 24 h after PTSD modelling, continuing for 14 days. Mice in the control group and model group were injected with an equal volume of 0.9% NaCl solution (once daily, for 14 days). The anxiety, despair, and learning and memory functions of the mice were assessed using the open field test, Y-maze test, and forced swimming test. Western blot was performed to measure the protein expression levels of methyltransferase-like protein 3 (METTL3), fat mass and obesity-associated gene (FTO), ALKB homolog 5 (ALKBH5), Wilms tumour 1 associating protein (WTAP), and methyltransferase-like protein 14 (METTL14) in the hippocampus. Immunofluorescence was used to detect the expression levels of METTL3, FTO, and ALKBH5 in the hippocampus. Statistical analysis was performed using SPSS 26.0 and GraphPad Prism 9.0.Comparisons between two groups were conducted using independent samples t-test, while comparisons among three groups were performed using one-way analysis of variance (ANOVA) or Kruskal-Wallis H test, followed by pairwise comparisons using LSD test. Results:(1) Behavioral results showed that the total distance travelled in the central area ( F=9.231, P<0.05) and the time spent in the central area ( H=8.045, P<0.05) showed statistically significant differences among the control, model, and sertraline groups. Mice in the control and sertraline groups travelled a greater distance((332.68±121.17)cm, (248.56±40.21)cm) and spent more time(24.98(23.08, 26.71)s, 22.52(18.86, 26.20)s) in the central area than those in the model group((131.66±84.90)cm, 9.14(6.56, 18.53)s) (all P<0.05). In the forced swimming test, the number of resting episodes ( F=16.882, P<0.05) and the duration of rest ( H=12.285, P<0.05) differed significantly among the three groups. Mice in the control group ((19.14±8.30) counts, 30.21 (18.98, 52.62) s) and the sertraline group ((17.63±8.14) counts, 25.90 (16.78, 37.56) s) had fewer resting episodes and shorter resting durations compared to those in the model group ((37.75±6.47) counts, 83.37 (64.62, 124.42) s) (all P<0.05). The percentage of alternations in the Y-maze experiment showed significant statistical differences among the three groups( F=6.844, P<0.05). Mice in the control group ((51.33±11.49)%) and the sertraline group ((48.24±3.10)%) exhibited a higher percentage of alternations than that in the model group ((36.70±8.15)%) ( P<0.05). (2) Western blot results showed that the protein expression levels of METTL3, FTO, and ALKBH5 in the hippocampal tissue of the three groups showed significant differences ( F=10.263, 9.010, 6.950, all P<0.05). The METTL3 and FTO protein expression levels in the hippocampus in the control group (0.85±0.07, 0.86±0.04) and the sertraline group (0.93±0.06, 0.95±0.13) were higher than those in the model group (0.74±0.02, 0.68±0.04) (all P<0.05). However, the ALKBH5 protein expression levels in the control group (0.93±0.08) and the sertraline group (0.87±0.13) were lower than that in the model group (1.13±0.04) (both P<0.05). (3) Immunofluorescence results showed that the expression levels of METTL3, FTO, and ALKBH5 proteins in the hippocampal tissue of the three groups showed significant statistical differences ( F=37.912, 62.659, 54.417, all P<0.05). The expression levels of METTL3 and FTO in the hippocampus in the control group (14.03±0.32, 13.85±0.28) and the sertraline group (17.94±0.29, 10.52±0.66) were higher than those in the model group (11.67±1.48, 8.70±0.68) (all P<0.05). The expression levels of ALKBH5 in the control group (12.94±0.38) and the sertraline group (13.30±0.93) were lower than that in the model group (19.24±1.03) (both P<0.05). Conclusion:The expression of m6A-related proteins in the hippocampus of PTSD-like mice is altered. Sertraline treatment can significantly regulate the expression of these proteins and improve anxiety, despair, and learning and memory impairments in the PTSD-like mice.
7.Analysis of immune infiltration mechanism of dermatomyositis and prediction of therapeutic targets of traditional Chinese medicine based on CIBERSORT algorithm
Pu WANG ; Min HU ; Suyue PAN ; Qiao HUANG ; Dongyu CHEN ; Wenlong FAN ; Xiaoyu YANG ; Hong-xin WANG ; Yuqing HE
Chinese Journal of Immunology 2025;41(4):783-791,中插1
Objective:To analyze the gene chip related to dermatomyositis based on bioinformatics,to explore the immune in-filtration mechanism of key genes in dermatomyositis by CIBERSORT deconvolution algorithm,and to predict the therapeutic targets of dermatomyositis by network pharmacology.Methods:The gene microarray of dermatomyositis was searched in GEO database,and the differentially coexpressed genes were screened and analyzed.The differentially coexpressed genes were analyzed by GO analysis,KEGG analysis,protein interaction network construction(PPI)by R software package.Verify the expression levels of key genes,and the correlation of immune cell infiltration was analyzed by CIBERSORT deconvolution method.Through the medical ontology informa-tion retrieval platform Coremine medical database,the traditional Chinese medicine treatment targets of dermatomyositis were screened and summarized.Results:A total of 196 differentially expressed genes were screened.GO enrichment analysis showed that these differentially expressed genes were mainly concentrated in defense response to virus,blood particles,double-stranded RNA binding,polypeptide antigen binding,and so on.KEGG enrichment analysis showed that it was enriched in RIG-Ⅰ-like receptor sig-nal pathway,Toll-like receptor signal pathway and other signal pathways related to the pathogenesis of dermatomyositis.Finally,four key genes of dermatomyositis,STAT1,ISG15,IRF7 and IRF9 were obtained.Through CIBERSORT algorithm,M1 macrophages,M2 macrophages and CD8+T cells were the three kinds of cells with the highest average proportion and the most obvious immune infil-tration,and there was a significant positive correlation between activated natural killer cells and activated dendritic cells,while there was a significant negative correlation between resting mast cells and activated mast cells.The therapeutic targets of traditional Chinese medicine such as fish brain stone were predicted based on Coremine medical database;through channel analysis,it could be found that these traditional Chinese medicines are mainly attributed to liver meridian,lung meridian,spleen meridian;efficacy analysis is mainly focused on clearing heat,detoxification,promoting blood circulation and removing blood stasis,relieving cough and resolving phlegm and so on.Conclusion:Four key genes and some key signal pathways of dermatomyositis,STAT1,ISG15,IRF7 and IRF9 were obtained by bioinformatics method,the immune infiltration mechanism was analyzed by CIBERSORT algorithm,and the thera-peutic potential targets of traditional Chinese medicine were screened out to provide direction for the pathogenesis and treatment of der-matomyositis.
8.The general pattern analysis of cirrhosis treatment based Ren-yin year's cases
Yirong HU ; Yixuan LIU ; Wenlong ZHENG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2025;32(4):448-453
Objective A data mining method was used to analyse and summarize Professor Zheng Wenlong's ideas and prescription and medication rules for the treatment of cirrhosis,taking the case of cirrhosis of the liver in Ren-yin year(2022)as an example.Methods A collection of traditional Chinese medicine prescriptions for liver cirrhosis patients who received treatment at the clinic of chief traditional Chinese medicine physician Zheng Wenlong at Hangzhou Traditional Chinese Medicine Hospital of Zhejiang Chinese Medical University from January 20,2022 to January 19,2023,and who met the inclusion and exclusion criteria.An excel database was created to standardize the data,followed by frequency statistical analyzes of prescriptions,medications and the characteristics of herbal remedies.Association rule analysis was carried out using SPSS Modeler 18,which facilitated the creation of complex network diagrams,subsequently enhanced through visualization in software Cytoscape.Clustering analysis was executed utilizing SPSS 25.0.Results A total of 176 prescriptions were selected,and the top 3 most frequently used prescriptions were Lingzhu decoction,Shengming decoction and Beihua decoction,with the frequency of 10.13%(23/227),10.13%(23/227)and 9.69%(22/227).A diverse array of 155 herbal medicines were employed,and the top 3 most frequently used drugs were Glycyrrhiza,Poria and Zingiberis Rhizoma Recens,with the frequency of 4.75%(127/2 676),4.26%(114/2 676)and 3.21%(86/2 676).The predominant medicinal properties of these herbs were classified as either cold or warm,while the primary tastes identified were bitter,sweet and spicy,with channel tropism were liver,lung,stomach and spleen.According to the descending order of improvement percentage,the results showed that the strongly correlated pairs of herbs were Chaenomelis Fructus-Aconiti Lateralis Radix Praeparata and Citri Reticulatae Pericarpium Viride-Pericarpium Citri Reticulatae,with improvement degrees of 2.74%and 2.55%respectively;the strongly correlated 3-herb combinations(Jiao Yao)included Aconiti Lateralis Radix Praeparata-Poria-Chaenomelis Fructus and Aconiti Lateralis Radix Praeparata-Rhizoma Atractylodis Macrocephalae-Chaenomelis Fructus,with improvement degrees of 3.24%and 3.23%respectively.The clustering analysis categorized the high-frequency herbs into 3 distinct groups:C1:Citri Reticulatae Pericarpium Viride,Pericarpium Citri Reticulatae,Jujubae Fructus,Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine,and Zingiberis Rhizoma Recens;C2:Rhizoma Atractylodis Macrocephalae,Aconiti Lateralis Radix Praeparata,Artemisiae Scopariae Herba,and Paeoniae Radix Alba;C3:Schisandra Chinensis,Rhizoma Alismatis,Radix Glycyrrhizae,Poria,Zingiberis Rhizoma and Ramulus Cinnamomi.These correspond to the formulas of Erchen decoction,Yinchenshufu decoction and Wuling powder.Conclusions The underlying pathogenesis of cirrhosis is linked to dysfunctions of the Jueyin and Yangming meridians,as well as disturbances in the Shaoyang axis.The primary therapeutic approach of Professor Zheng Wenlong involves a comprehensive evaluate all of the patient's clinical presentation to identify the shared pathogenesis underlying various symptoms,integrating the patient's personal constitution with the annual cycle of the Five Phases(Wuyun)and Six Qi(Liuqi)as described in traditional Chinese medicine,and formulate prescriptions based on the interactions of the Five Phases(Wuxing)and guided by the yin-yang theory of"Opening-Closing-Pivot"(Kai-He-Shu).The focus of prescriptions is rehabilitation and the final goal is reach the state of"Tian Ren He Yi".
9.Analysis of immune infiltration mechanism of dermatomyositis and prediction of therapeutic targets of traditional Chinese medicine based on CIBERSORT algorithm
Pu WANG ; Min HU ; Suyue PAN ; Qiao HUANG ; Dongyu CHEN ; Wenlong FAN ; Xiaoyu YANG ; Hong-xin WANG ; Yuqing HE
Chinese Journal of Immunology 2025;41(4):783-791,中插1
Objective:To analyze the gene chip related to dermatomyositis based on bioinformatics,to explore the immune in-filtration mechanism of key genes in dermatomyositis by CIBERSORT deconvolution algorithm,and to predict the therapeutic targets of dermatomyositis by network pharmacology.Methods:The gene microarray of dermatomyositis was searched in GEO database,and the differentially coexpressed genes were screened and analyzed.The differentially coexpressed genes were analyzed by GO analysis,KEGG analysis,protein interaction network construction(PPI)by R software package.Verify the expression levels of key genes,and the correlation of immune cell infiltration was analyzed by CIBERSORT deconvolution method.Through the medical ontology informa-tion retrieval platform Coremine medical database,the traditional Chinese medicine treatment targets of dermatomyositis were screened and summarized.Results:A total of 196 differentially expressed genes were screened.GO enrichment analysis showed that these differentially expressed genes were mainly concentrated in defense response to virus,blood particles,double-stranded RNA binding,polypeptide antigen binding,and so on.KEGG enrichment analysis showed that it was enriched in RIG-Ⅰ-like receptor sig-nal pathway,Toll-like receptor signal pathway and other signal pathways related to the pathogenesis of dermatomyositis.Finally,four key genes of dermatomyositis,STAT1,ISG15,IRF7 and IRF9 were obtained.Through CIBERSORT algorithm,M1 macrophages,M2 macrophages and CD8+T cells were the three kinds of cells with the highest average proportion and the most obvious immune infil-tration,and there was a significant positive correlation between activated natural killer cells and activated dendritic cells,while there was a significant negative correlation between resting mast cells and activated mast cells.The therapeutic targets of traditional Chinese medicine such as fish brain stone were predicted based on Coremine medical database;through channel analysis,it could be found that these traditional Chinese medicines are mainly attributed to liver meridian,lung meridian,spleen meridian;efficacy analysis is mainly focused on clearing heat,detoxification,promoting blood circulation and removing blood stasis,relieving cough and resolving phlegm and so on.Conclusion:Four key genes and some key signal pathways of dermatomyositis,STAT1,ISG15,IRF7 and IRF9 were obtained by bioinformatics method,the immune infiltration mechanism was analyzed by CIBERSORT algorithm,and the thera-peutic potential targets of traditional Chinese medicine were screened out to provide direction for the pathogenesis and treatment of der-matomyositis.
10.Expression of m6A-related proteins in mice with PTSD-like behavior improved by sertraline hydrochloride treatment
Jiaying LU ; Luodong YANG ; Min HU ; Wenlong XING ; Haiteng CUI ; Guiqing ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(11):961-968
Objective:To explore the expression changes of N6-methyladenosine (m6A)-related proteins in the hippocampus of mice with post traumatic stress disorder (PTSD)-like behavior and the therapeutic effects of sertraline hydrochloride.Methods:Male C57BL/6J mice aged 4-6 weeks were selected to establish a PTSD model using a single prolonged stress and foot shock stimulation. A total of 24 mice were randomly divided into the control group, model group, and sertraline group using a random number table, with 8 mice in each group. Mice in the sertraline group were intraperitoneally injected with sertraline hydrochloride (15 mg/kg, once daily) 24 h after PTSD modelling, continuing for 14 days. Mice in the control group and model group were injected with an equal volume of 0.9% NaCl solution (once daily, for 14 days). The anxiety, despair, and learning and memory functions of the mice were assessed using the open field test, Y-maze test, and forced swimming test. Western blot was performed to measure the protein expression levels of methyltransferase-like protein 3 (METTL3), fat mass and obesity-associated gene (FTO), ALKB homolog 5 (ALKBH5), Wilms tumour 1 associating protein (WTAP), and methyltransferase-like protein 14 (METTL14) in the hippocampus. Immunofluorescence was used to detect the expression levels of METTL3, FTO, and ALKBH5 in the hippocampus. Statistical analysis was performed using SPSS 26.0 and GraphPad Prism 9.0.Comparisons between two groups were conducted using independent samples t-test, while comparisons among three groups were performed using one-way analysis of variance (ANOVA) or Kruskal-Wallis H test, followed by pairwise comparisons using LSD test. Results:(1) Behavioral results showed that the total distance travelled in the central area ( F=9.231, P<0.05) and the time spent in the central area ( H=8.045, P<0.05) showed statistically significant differences among the control, model, and sertraline groups. Mice in the control and sertraline groups travelled a greater distance((332.68±121.17)cm, (248.56±40.21)cm) and spent more time(24.98(23.08, 26.71)s, 22.52(18.86, 26.20)s) in the central area than those in the model group((131.66±84.90)cm, 9.14(6.56, 18.53)s) (all P<0.05). In the forced swimming test, the number of resting episodes ( F=16.882, P<0.05) and the duration of rest ( H=12.285, P<0.05) differed significantly among the three groups. Mice in the control group ((19.14±8.30) counts, 30.21 (18.98, 52.62) s) and the sertraline group ((17.63±8.14) counts, 25.90 (16.78, 37.56) s) had fewer resting episodes and shorter resting durations compared to those in the model group ((37.75±6.47) counts, 83.37 (64.62, 124.42) s) (all P<0.05). The percentage of alternations in the Y-maze experiment showed significant statistical differences among the three groups( F=6.844, P<0.05). Mice in the control group ((51.33±11.49)%) and the sertraline group ((48.24±3.10)%) exhibited a higher percentage of alternations than that in the model group ((36.70±8.15)%) ( P<0.05). (2) Western blot results showed that the protein expression levels of METTL3, FTO, and ALKBH5 in the hippocampal tissue of the three groups showed significant differences ( F=10.263, 9.010, 6.950, all P<0.05). The METTL3 and FTO protein expression levels in the hippocampus in the control group (0.85±0.07, 0.86±0.04) and the sertraline group (0.93±0.06, 0.95±0.13) were higher than those in the model group (0.74±0.02, 0.68±0.04) (all P<0.05). However, the ALKBH5 protein expression levels in the control group (0.93±0.08) and the sertraline group (0.87±0.13) were lower than that in the model group (1.13±0.04) (both P<0.05). (3) Immunofluorescence results showed that the expression levels of METTL3, FTO, and ALKBH5 proteins in the hippocampal tissue of the three groups showed significant statistical differences ( F=37.912, 62.659, 54.417, all P<0.05). The expression levels of METTL3 and FTO in the hippocampus in the control group (14.03±0.32, 13.85±0.28) and the sertraline group (17.94±0.29, 10.52±0.66) were higher than those in the model group (11.67±1.48, 8.70±0.68) (all P<0.05). The expression levels of ALKBH5 in the control group (12.94±0.38) and the sertraline group (13.30±0.93) were lower than that in the model group (19.24±1.03) (both P<0.05). Conclusion:The expression of m6A-related proteins in the hippocampus of PTSD-like mice is altered. Sertraline treatment can significantly regulate the expression of these proteins and improve anxiety, despair, and learning and memory impairments in the PTSD-like mice.

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