1.Treatment of Sepsis-induced Inflammatory Responses with Xijiao Dihuangtang by Modulation of PKM2-mediated One-carbon Metabolism Pathway
Qixiang YAN ; Yeyan ZHU ; Fan GE ; Qimeng SUN ; Leyao YE ; Fang TIAN ; Jun LU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):18-26
ObjectiveTo investigate the effects of Xijiao Dihuangtang (XJDHT) on mice with sepsis and cellular models of sepsis and explore its molecular mechanism in alleviating sepsis-induced inflammatory responses via regulating pyruvate kinase M2 (PKM2)-mediated one-carbon metabolism pathway. MethodsForty C57BL/6N mice were randomly divided into four groups: normal group, model group, low-dose XJDHT group (7.7 g·kg-1), and high-dose XJDHT group (15.4 g·kg-1). After one week of continuous gavage, sepsis was induced using cecal ligation and puncture (CLP) in groups except the normal group. 24 h after the surgery, mortality rates in all groups were recorded, and serum cytokines were measured by enzyme linked immunosorbent assay (ELISA). Lung histopathology was examined by hematoxylin-eosin (HE) staining. During the in vitro experiment, the human monocytic leukemia cell line (THP-1) was exposed to various concentrations of XJDHT and treated with lipopolysaccharide (LPS) at a final concentration of 2 mg·L-1 for 24 h. Cell apoptosis was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Protein levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were measured by Western blot. Transcriptome sequencing was performed to analyze differentially expressed genes in all groups and conduct gene ontology (GO) enrichment. Key genes in the one-carbon metabolism pathway, including pyruvate kinase M2 (PKM2), 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR), and phosphoglycerate dehydrogenase (PHGDH), were verified by Western blot. A PKM2 inhibition model was established using shikonin for further protein expression analysis. ResultsAnimal experiments showed that compared with the normal group, the model group exhibited significantly elevated body temperature and lung pathology (P<0.01) and increased serum TNF-α and IL-1β levels (P<0.01). High-dose XJDHT reduced body temperature and lung tissue damage (P<0.01) and significantly decreased serum TNF-α and IL-1β levels (P<0.01). Low-dose XJDHT treatment showed no significant temperature change (P<0.01) but reduced serum TNF-α and IL-1β levels (P<0.01). Transcriptome sequencing and Western blot revealed significant differences in the expression of TNF-α, IL-1β, and one-carbon metabolism genes (PKM2, MTR, and PHGDH) (P<0.01). Cell experiments demonstrated that compared to the normal group, the model group showed elevated protein expressions of TNF-α and IL-1β in THP-1 cells (P<0.01), decreased Bcl-2/Bax ratio, and increased apoptosis (P<0.01). Transcriptome sequencing and Western blot revealed significant differences in the expression of TNF-α, IL-1β, and one-carbon metabolism genes (PKM2, MTR, and PHGDH) (P<0.01). Compared to the model group, high-dose XJDHT significantly increased Bax/Bcl-2 ratio and PHGDH protein expression (P<0.01) and effectively reduced cell apoptosis (P<0.01) while down-regulating protein expressions of TNF-α, IL-1β, PKM2, and MTR (P<0.01). Low-dose XJDHT moderately increased Bax/Bcl-2 ratio and PHGDH protein expression (P<0.05), reduced apoptosis (P<0.05), and decreased IL-1β and MTR protein levels (P<0.05, P<0.01), but there were no significant changes in TNF-α and PKM2 expression. After PKM2 inhibition by shikonin in THP-1 cells, the expression of protein related to one-carbon metabolism was detected. Compared with the blank group, the LPS-induced model group showed significantly upregulated PKM2 and MTR protein expression (P<0.01) and downregulated PHGDH expression (P<0.01). Compared with the model group, shikonin treatment significantly reduced PKM2 expression (P<0.05), increased PHGDH expression (P<0.01), and decreased MTR expression (P<0.05). ConclusionXJDHT can inhibit the release of inflammatory factors in sepsis, and its mechanism is related to the intervention of the PKM2-regulated one-carbon metabolism pathway in macrophages.
2.Exploration of mechanism of action of tretinoin polyglucoside in rats with IgA nephropathy based on mitochondrial dynamics
Yan-Min FAN ; Shou-Lin ZHANG ; Hong FANG ; Xu WANG ; Han-Shu JI ; Ji-Chang BU ; Ke SONG ; Chen-Chen CHEN ; Ying DING ; Chun-Dong SONG
Chinese Pharmacological Bulletin 2024;40(11):2069-2074
Aim To investigate the effects of multi-gly-cosides of Tripterygium wilfordii(GTW)on mitochon-drial dynamics-related proteins and the mechanism of nephroprotective effects in IgA nephrophathy(IgAN)rats.Methods SPF grade male SD rats were random-ly divided into the Control group,modelling group,prednisone group(6.25 mg·kg·d-1)and GTW group(6.25 mg·kg·d-1).The IgAN rat model was established by the method of"bovine serum albumin(BSA)+carbon tetrachloride(CCl4)+lipopolysac-charide(LPS)".The total amount of urinary protein(24 h-UTP)and erythrocyte count in urine were meas-ured in 24 h urine.Blood biochemistry of serum albu-min(ALB),alanine aminotransferase(ALT),urea ni-trogen(BUN),and creatinine(Scr)were measured in abdominal aorta of the rats;immunofluorescence and HE staining were used to observe the histopathology of the kidneys;RT-PCR and Western blotting were used to detect the mRNA and protein expression levels of key proteins regulating mitochondrial division and fu-sion:dynamin-related protein 1(Drp1),mitochondrial fusion protein 1(Mfn1),and mitochondrial fusion pro-tein 2(Mfn2),and PTEN-induced putative kinase 1(Pink1),in the kidney tissue of rats.Results GTW significantly reduced urinary erythrocyte count and 24 h-UTP,decreased serum ALT,BUN and Scr levels,in-creased serum ALB levels,improved renal histopatho-logical status in IgAN rats,increased the protein and mRNA expression levels of Mfn1,Mfn2,and Pink1,and decreased the protein and mRNA expression levels of Drp1 in renal tissues.Conclusions GTW may regu-late mitochondrial structure and maintain the dynamic balance of mitochondrial dynamics by promoting the ex-pression of Mfn1,Mfn2,Pink1 and decreasing Drp1.This may result in a reduction in urinary erythrocyte counts and proteinuria,and an improvement in renal function.
3.Effects of Tripterygium glycosides tablets on LIGHT-HVEM/LTβR pathway in rats with IgA nephropathy
Xu WANG ; Hong FANG ; Yan-Min FAN ; Han-Shu JI ; Ke SONG ; Chen-Chen CHEN ; Ji-Chang BU ; Ying DING ; Chun-Dong SONG
Chinese Pharmacological Bulletin 2024;40(12):2277-2282
Aim To explore the mechanism of action of Tripterygium glycosides tablets on kidney of rats with IgA nephropathy based on inflammation-related path-ways.Methods Forty-five male SD rats of SPF grade were randomly divided into control group and modeling group.In addition to the blank group,the modeling group used the combination of bovine serum albumin(BSA)+carbon tetrachloride(CC14)+lipopolysac-charide(LPS)to establish the IgA nephropathy rat model.Successfully modeled rats were randomly divid-ed into the model group,the prednisone group and Tripterygium glycosides tablets group,and the treat-ment group was given the drug by gavage from the 13 th week,and the 24 hours urine,blood and kidney tis-sues of the rats were collected and examined after 4 weeks of the administration of the drug.Urine erythro-cyte count,quantitative 24-h urine protein(24 h-UTP),urea nitrogen(BUN),and blood creatinine(Scr)were detected in each group;serum interleukin 1β(IL-1β)and tumor necrosis factor α(TNF-α)were detected by enzyme-linked immunosorbent assay(Elisa);the pathological changes in the renal tissues of the rats in each group were observed by horizontal hematoxylin-eosin(HE)staining;and the renal tis-sues in each group were observed by Western blotting.The expressions of LIGHT,HVEM,LTβR proteins and their mRNAs in rat kidney tissue were detected by Western blot and real-time fluorescence quantitative polymerase chain reaction(RT-PCR).Results Tripterygium glycosides tablets significantly reduced the levels of urinary erythrocyte count,24 h-UTP,BUN,and Scr in IgA nephropathy rats(P<0.01),improved renal histopathology,lowered the levels of se-rum inflammatory factors IL-1β and TNF-α(P<0.01),and lowered the levels of LIGHT,HVEM,LTβR proteins and their mRNA expression in renal tis-sues(P<0.01).Conclusions Tripterygium glyco-sides tablets may inhibit the immune response and re-duce the release of inflammatory factors by down-regu-lating the LIGHT-HVEM/LT(3R pathway,thus reduc-ing the inflammatory response,lowering the urinary e-rythrocytes and urinary proteins,improving the renal nephron pathologic injury,and protecting the renal function.
4.Summary of best evidence for sexual health management in colorectal cancer patients
Jinghui FAN ; Lili ZHANG ; Siqin YAN ; Chengyuan FANG ; Gaohua WANG
Chinese Journal of Modern Nursing 2024;30(21):2837-2843
Objective:To integrate the best evidence of sexual health management for colorectal cancer patients both domestically and internationally, so as to provide guidance for clinical practice.Methods:Evidence on sexual health management of colorectal cancer patients in domestic and foreign databases and websites was systematically searched, and evidence from literature that met quality evaluation standards was extracted and summarized. The search period was from July 2003 to July 2023.Results:A total of 16 articles were included, including five clinical decisions, six clinical practice guidelines, three expert consensus/group standards, and two systematic reviews. Thirty-four recommended opinions were formed, including eight aspects of evaluation and screening, assessment tools, manifestations of sexual dysfunction, coping strategies for sexual problems, psychological intervention, measures for fertility protection, health education, and follow-up management.Conclusions:This study summarizes the best evidence for sexual health management in colorectal cancer patients. It is necessary to selectively apply evidence based on the evaluation of patient conditions by medical and nursing staff and clinical situations to ensure effective and scientific sexual health management strategies for patients, and promote the translation of best evidence into clinical practice.
5.Evaluation and optimization of metagenomic sequencing platforms for bloodstream infection samples
Xin PENG ; Hang FAN ; Meng-Nan CUI ; Lei LIN ; Guang-Qian PEI ; Yun-Fei WANG ; Xiu-Juan ZUO ; Xiao-Feng FANG ; Yan GUO ; Yu-Jun CUI
Chinese Journal of Zoonoses 2024;40(10):928-934
This study was aimed at comparing performance differences among three metagenomic sequencing platforms,MGISEQ-2000,Illumina NextSeq 2000,and Ion GeneStudio S5 Plus,to optimize the sequencing process for trace samples.The three sequencing platforms were used to perform high-throughput sequencing on DNA standards and simulated samples.Through analysis of the quality of raw data and microbial detection capabilities,systematic differences among platforms were compared.The sequencing results were optimized for trace samples by incorporation of exogenous nucleic acids during the li-brary preparation process.In terms of data output per batch and base quality,MGISEQ-2000 surpassed the other two plat-forms.Illumina NextSeq 2000 had the lowest proportion of duplicate reads,whereas Ion GeneStudio S5 Plus had the highest proportion,and significant differences were observed across platforms(P<0.001).In sequencing uniformity,MGISEQ-2000 and Illumina NextSeq 2000 were superior to Ion GeneStudio S5 Plus.MGISEQ-2000 provided a substantial advantage in microbial detection capability(P<0.001),but the advantage diminished with decreasing bacterial fluid concentration.Ion GeneStudio S5 Plus had the shortest duration for single-batch sequencing.Moreo-ver,for trace samples with DNA content ≤0.05 ng,the experi-mental group(with added exogenous nucleic acids)achieved a higher number of reads than the control group(without exogenous nucleic acids),with a 11.09±8.03 fold increase.In conclu-sion,the different sequencing platforms each had advantages and disadvantages,thus allowing researchers to choose the appro-priate platform according to specific needs.Furthermore,the addition of exogenous nucleic acids improved the microorganism detection efficiency,and provided better support for subsequent diagnosis and evaluation of results.
6.C/EBPα Positively Regulates Chicken NRG4 Gene Expression via Binding to Its Exon 2
Hao-Yu LUO ; Jia-Xin HUANG ; Fan GAO ; Xiao-Hong YAN ; Fang MU ; Ning WANG
Chinese Journal of Biochemistry and Molecular Biology 2024;40(11):1608-1617
Neuregulin 4 (NRG4) is a newly discovered adipokine,which plays crucial roles in many physiological processes such as energy balance,and glucose and lipid metabolism.The transcriptional regulation of NRG4 gene remains largely unknown.Our previous 5'RACE analysis showed that the tran-scription start sites (TSS) of chicken NRG4 gene were dispersed over a region of 200 bp,suggesting that chicken NRG4 gene possesses a dispersed promoter.In the present study,the promoter activity of the proximal exon and intron of chicken NRG4 gene was analyzed.Dual-luciferase reporter assay demonstra-ted that the genomic fragment (-122/+452) containing exon 1,intron 1,and exon 2 of chicken NRG4 gene had strong bidirectional promoter activity.Bioinformatics analysis showed that there was a putative binding site of the transcription factor CCAAT enhancer-binding proteinα(C/EBPα) in exon 2 of chick-en NRG4 gene.Reporter gene assay showed that both deletion and site-directed mutagenesis of the C/EBPα binding site abrogated the regulatory effect of C/EBPα on the promoter activity of the-122/+452 fragment of chicken NRG4 gene.Gene expression correlation analysis showed that C/EBPα and NRG4 gene expression were positively correlated in chicken adipose tissues.ChIP-PCR revealed that C/EBPα directly binds to the exon 2 of NRG4 gene in chicken adipose.In conclusion,the proximal exon and intron of chicken NRG4 gene has bidirectional promoter activity,and C/EBPα directly regulates NRG4 gene expression in chicken adipose via binding to its exon 2.
7.Clinical Features and Prognosis of Patients with CD5+Diffuse Large B-Cell Lymphoma
Xiu-Juan HUANG ; Jian YANG ; Xiao-Fang WEI ; Yuan FU ; Yang-Yang ZHAO ; Ming-Xia CHENG ; Qing-Fen LI ; Hai-Long YAN ; You-Fan FENG
Journal of Experimental Hematology 2024;32(3):750-755
Objective:To analyze the clinical characteristics and prognosis of patients with CD5+diffuse large B-cell lymphoma(DLBCL).Methods:The clinical data of 161 newly treated DLBCL patients in Gansu Provincial Hospital from January 2013 to January 2020 were retrospectively analyzed.According to CD5 expression,the patients were divided into CD5+group and CD5-group.The clinical characteristics and prognosis of the two groups were statistically analyzed.Results:The median age of patients in CD5+group was 62 years,which was higher than 56 years in CD5-group(P=0.048).The proportion of women in CD5+group was 62.96%,which was significantly higher than 41.79%in CD5-group(P=0.043).The proportion of patients with IPI score>2 in CD5+group was 62.96%,which was higher than 40.30%in CD5-group(P=0.031).Survival analysis showed that the median overall survival and progression-free survival time of patients in CD5+group were 27(3-77)and 31(3-76)months,respectively,which were both shorter than 30(5-84)and 32.5(4-83)months in CD5-group(P=0.047,P=0.026).Univariate analysis showed that advanced age,positive CD5 expression,triple or double hit at initial diagnosis,high IPI score and no use of rituximab during chemotherapy were risk factors for the prognosis of DLBCL patients.Further Cox multivariate regression analysis showed that these factors were also independent risk factors except for advanced age.Conclusion:CD5+DLBCL patients have a worse prognosis than CD5-DLBCL patients.Such patients are more common in females,with advanced age and high IPI score,which is a special subtype of DLBCL.
8.Analysis of Incidence Rate,Risk Factors and Prognosis of Pulmo-nary Hypertension in Ph-MPNs Patients
Hong-Xia AN ; Qi-Ke ZHANG ; Xiao-Fang WEI ; You-Fan FENG ; Yan-Qing SUN
Journal of Experimental Hematology 2024;32(5):1463-1471
Objective:To explore and analyze the incidence rate,influencing factors and impact on prognosis of pulmonary hypertension(PH)in patients with Philadelphia chromosome negative myeloproliferative neoplasms(Ph-MPNs).Methods:The clinical data of 271 patients with Ph-MPNs were retrospectively analyzed,and different disease subtypes were classified.Patients with different disease types were further divided into PH+and PH-groups according to whether HP occurred.Statistical methods were used to analyze the incidence rate,risk factors,and impact on prognosis of PH in Ph-MPNs patients.Results:The overall incidence rate of PH among 271 patients was 26.9%,and according to the classification of disease subtypes,it was found that the incidence rate of PH in patients with primary myelofibrosis(PMF)was significantly higher than those of patients with polycythemia vera and essential thrombocythemia(both P<0.05).Multivariate regression analysis showed that advanced age,long disease course,JAK2 positive and increased hematocrit,lactate dehydrogenase,monocyte count,and uric acid level were independent risk factors for PH in Ph-MPNs patients(OR>1,P<0.05),and there were some differences in the independent risk factors between different disease subtypes.Survival analysis results showed that the overall survival(OS)rate of PH+patients was significantly lower than that of PH-patients in other types except for PMF(all P<0.05).Conclusion:The incidence rate of PH in Ph-MPNs patients is high,and its risk factors are diverse.The OS rate of Ph-MPNs patients with PH is low.Therefore,we should be highly alert to the occurrence of PH in Ph-MPNs patients clinically.
9.CD163 expression characteristics in tumor tissue and blood of patients with intracranial aneurysm and its use as a molecular marker
Yuanfeng WEN ; Quan YAN ; Guofeng FAN ; Peng CHENG ; Taian FANG ; Gonggaoang BATE ; Xuefei LIU ; Yun WANG ; Guojia DU
Chinese Journal of Neuromedicine 2024;23(5):464-470
Objective:To investigate the CD163 expression characteristics in intracranial aneurysm (IA) tissue and blood of patients with IA and its feasibility as an early clinical screening indicator for IA.Methods:A total of 28 patients with IA admitted to Department of Neurosurgery, First Affiliated Hospital of Xinjiang Medical University from January 2021 to November 2023 were selected as IA group, and 28 healthy subjects from Health Management Center, First Affiliated Hospital of Xinjiang Medical University at the same time period were selected as control group. Eight saccular IA tissues and 12 superficial temporal artery tissues were collected from patients from IA group accepted IA clipping, and real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the CD163 mRNA expression in these tissues. RT-qPCR was also used to detect the CD163 mRNA expression in the blood of the 2 groups. Seven patients with IA and 7 control subjects from the above 2 groups were randomly selected, respectively; and plasma CD163 protein content was detected by enzyme-linked immunosorbent assay (ELISA). Multivariate Logistic regression was used to analyze the influencing factors for IA. Receiver operating characteristic (ROC) curve was used to analyze the diagnostic values of blood CD163 mRNA expression and plasma CD163 protein content in IA. Results:CD163 mRNA expression in IA tissues was significantly higher than that in superficial temporal artery tissues (41.870±20.355 vs. 6.080±5.444, P<0.05). CD163 mRNA expression in the blood of IA patients was significantly higher than that in the controls (1.969[1.124, 2.318] vs. 1.124[0.933, 1.379], P<0.05). CD163 mRNA expression in the blood of ruptured IA group, unruptured IA group, and control group was gradually decreased, with significant differences ( P<0.05). CD163 mRNA expression in the blood of female IA patients was not statistically different compared with that in male IA patients ( P>0.05). ELISA showed that the CD163 protein content in plasma of the IA group was significantly higher than that in the control group [10.537±1.879] ng/L vs. [8.598±0.885] ng/L, P<0.05). Multivariate Logistic regression analysis showed that age and CD163 mRNA expression in the blood were independent influencing factors for IA occurrence ( OR=0.844, 95% CI: 0.750-0.951, P=0.005; OR=0.111, 95% CI: 0.024-0.506, P=0.004). ROC curve showed that the area under the curve (AUC) of CD163 mRNA expression in blood in diagnosing IA was 0.759 (95% CI: 0.618-0.890, P=0.002), and that of CD163 protein content in plasma in diagnosing IA was 0.864 (95% CI: 0.610-1.000, P=0.035). Conclusion:CD163 mRNA expressions in blood and IA tissues and CD163 protein content in plasma are high in patients with IA; CD163 mRNA expression in blood is an independent risk factor for IA; CD163 protein in plasma can be used as a molecular marker for screening IA.
10.Design of new type of acupressure wrist-ankle strap and its value for treating mild anxiety insomnia
Ying YUAN ; Wei-Hong LI ; Yan-Li YOU ; Qian-Xi DAI ; Fan-Fu FANG
Chinese Medical Equipment Journal 2024;45(4):78-82
Objective To design a new type of acupressure wrist-ankle strap and evaluate its clinical effect on the treatment of patients with mild anxiety insomnia.Methods The acupressure wrist-ankle strap was composed of a fixation band and an acupressure part.Totally 94 insomnia patients with mild anxiety at some hospital from October 2020 to January 2023 were selected as the subjects and divided into an observation group(n=48)and a control group(n=46)with the random number table method.The observation group was treated with the acupressure wrist-ankle strap,and the control group was given with placebos.The two groups were compared before and after treatment in terms of Pittsburgh sleep quality index(PSQI)and Hamilton anxiety scale(HAMA)with two-way repeated measures ANOVA.SPSS 21.0 software was used for data analysis.Results The PSQI and HAMA scores of the two groups were enhanced significantly after treatment(P<0.05).The two groups had statistically significant difference in the effect of the time factor on PSQI and HAMA scores(P<0.05),and also in the effect of the time x group interaction on PSQI and HAMA scores(P<0.05).Condusion The acupressure wrist-ankle strap contributes to improving the sleep and anxiety of mild anxiety insomnia patients in a self-administered,non-invasive,painless,and non-adverse manner.[Chinese Medical Equipment Journal,2024,45(4):78-82]

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