1.Mechanism of Herbal Cake-separated Moxibustion in Improving Neuroimmune Inflammation in Rats with Chronic Fatigue Syndrome by Interfering TLR4/MyD88/NF-κB Pathway
Chuntao ZHAI ; Yawei HOU ; Linjuan SHI ; Yixiao WANG ; Wei LI ; Yuefeng TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):140-149
ObjectiveTo explore the mechanism of herbal cake-separated moxibustion using the classical formula Xiaoyaosan in alleviating neuroimmune inflammatory responses in chronic fatigue syndrome (CFS) rats, based on the regulation of the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor-κB (NF-κB) signaling pathway. MethodsFifty SPF-grade SD rats aged 6-8 weeks were randomly divided into five groups: Normal group, model group, sham herbal cake moxibustion group, Chinese medicine intragastric administration group, and herbal cake-separated moxibustion group, with 10 rats in each group. Except for the normal group, all other groups underwent a 21-day modeling process, followed by behavioral testing. The herbal cake-separated moxibustion and sham herbal cake moxibustion groups received interventions at the Shenque (CV8), Guanyuan (CV4), Zusanli (ST36), and Qimen (LR14) acupoints. The Chinese medicine intragastric administration group was treated with a Xiaoyaosan suspension via gavage. Behavioral tests were conducted after 10 days of continuous intervention. Serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α), as well as hippocampal levels of IL-1β, IL-6, TNF-α, and NF-κB, were detected by enzyme-linked immunosorbent assay (ELISA). Morphological changes in the hippocampus were observed using hematoxylin-eosin (HE) staining. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect mRNA expression levels of TLR4, MyD88, and NF-κB in the hippocampus. Western blot analysis was performed to detect the relative expression levels of TLR4, MyD88, NF-κB, and p65 proteins in the hippocampus. ResultsCompared with the normal group, the model group showed a significant decrease in upright times during the open field test (P<0.01), as well as significant reductions in total movement distance, resting time, and center region duration (P<0.01). In the tail suspension test, immobility time increased (P<0.01), and struggle times decreased (P<0.01). Serum and hippocampal levels of IL-1β, IL-6, and TNF-α, as well as hippocampal NF-κB levels and TLR4, MyD88, and NF-κB mRNA expression, were significantly elevated (P<0.01). After treatment, compared with the model group, the total movement distance and upright times in the open field test were significantly increased in all treatment groups (P<0.01), while resting time and center region duration were notably prolonged (P<0.05, P<0.01). Immobility time in the tail suspension test was significantly shortened (P<0.01), and struggle times significantly increased (P<0.05). Serum and hippocampal levels of IL-1β, IL-6, TNF-α, hippocampal NF-κB levels, and TLR4 and NF-κB mRNA expression were significantly reduced (P<0.05, P<0.01). Compared with the sham herbal cake moxibustion group, the herbal cake-separated moxibustion group showed a significant extension in center region duration during the open field test (P<0.05) and a significant increase in upright times (P<0.01). In the tail suspension test, immobility time was reduced (P<0.01), and struggle times increased (P<0.01). Serum TNF-α levels in the Chinese medicine intragastric administration group were significantly reduced (P<0.01), while serum IL-6 levels, as well as hippocampal levels of IL-1β, TNF-α, NF-κB, and TLR4, MyD88, and NF-κB mRNA expression, were significantly decreased in both the Chinese medicine intragastric administration group and the herbal cake-separated moxibustion group (P<0.05, P<0.01). Compared with the Chinese medicine intragastric administration group, the herbal cake-separated moxibustion group exhibited significantly increased upright times in the open field test (P<0.01). In the tail suspension test, immobility time was reduced (P<0.01), and struggle times increased (P<0.01). Serum IL-1β, hippocampal TNF-α levels, and TLR4, MyD88, and NF-κB mRNA expression were significantly decreased (P<0.05, P<0.01). ConclusionHerbal cake-separated moxibustion effectively improves fatigue and memory function in CFS rats, regulates neuroimmune inflammatory responses, and its mechanism may be related to the modulation of the TLR4/MyD88/NF-κB signaling pathway.
2.Herbal Cake-separated Moxibustion Improves Cognitive Function in Rat Model of Chronic Fatigue Syndrome via PI3K/Beclin1 Signaling Pathway
Chuntao ZHAI ; Yawei HOU ; Linjuan SHI ; Yixiao WANG ; Yuefeng TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):149-158
ObjectiveTo explore the mechanism of moxibustion with the classical prescription Xiaoyaosan as the herbal cake in improving cognitive function in the rat model of chronic fatigue syndrome (CFS) by regulating autophagy via phosphatidylinosiol 3-kinase (PI3K)/autophagy key molecule yeast Atg6 homologue 1 (Beclin1) signaling pathway. MethodsFifty SPF-grade SD rats aged 6-8 weeks were randomly grouped as follows: control, model, sham cake-separated moxibustion, intragastric administration with Chinese herbs, and herbal cake-separated moxibustion, each with 10 rats. The other groups except the control group were subjected to modeling by exhaustive swimming and chronic restraint stress for 21 days, and the behavioral test was performed after modeling. Herbal cake-separated moxibustion and sham cake-separated moxibustion were carried out at Shenque (CV8), Guanyuan (CV4), Zusanli (ST36), and Qimen (LR14). The rats in the intragastric administration with Chinese herbs group were treated with Xiaoyaosan suspension by gavage. After continuous intervention for 10 days, rat behaviors were observed and the levels of interleukin (IL)-4, IL-10, blood urine nitrogen (BUN), and malondiadehyde (MDA) in the serum and hippocampal tissue of rats in each group were determined by enzyme-linked immunosorbent assay. The morphological changes of the hippocampal tissue were observed by hematoxylin-eosin staining. The relative mRNA and protein levels of PI3K and Beclin1 in the hippocampal tissue were determined by Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. ResultsCompared with the normal group, the model group showed reduced crossings, shortened residence time in the target quadrant, and prolonged average escape latency on days 1, 2, 3, and 4 (P<0.01) in the Morris water maze test. In the new object recognition test, the model group showed decreased recognition index of exploration time and recognition index of exploration times (P<0.01), lowered levels of IL-4 and IL-10 and elevated levels of BUN and MDA in the serum and hippocampus (P<0.01), and down-regulated mRNA and protein levels of PI3K and Beclin1 (P<0.01). After treatment, compared with the model group, each treatment group showed increased crossings, prolonged residence time in the target quadrant, and shortened average escape latency on days 1, 2, 3, and 4 (P<0.05, P<0.01). In addition, the treatment elevated the levels of IL-4 and IL-10 and lowered the levels of BUN and MDA in the serum and hippocampus (P<0.05, P<0.01), and up-regulated the mRNA and protein levels of PI3K and Beclin1 (P<0.05, P<0.01). Compared with the sham cake-separated moxibustion group, the herbal cake-separated moxibustion group and the intragastric administration with Chinese herbs group showed increased crossings, prolonged residence time in the target quadrant (P<0.05), elevated level of IL-4, lowered level of MDA, and up-regulated relative mRNA and protein levels of PI3K and Beclin1 in the hippocampus (P<0.05, P<0.01). The serum levels of MDA and BUN in the intragastric administration with Chinese herbs group were significantly decreased. The average escape latency of the herbal cake-separated moxibustion group was significantly shortened on days 2 and 3 (P<0.05, P<0.01). In addition, this group showed elevated serum level of IL-4, lowered serum level of MDA, and declined BUN level in the hippocampus (P<0.05, P<0.01). Compared with the intragastric administration with Chinese herbs group, the herbal cake-separated moxibustion group presented risen serum level of IL-4 and up-regulated mRNA level of Beclin1 (P<0.05) and relative protein levels of PI3K and Beclin1 (P<0.05, P<0.01). ConclusionHerbal cake-separated moxibustion can effectively alleviate fatigue and improve the memory, reduce inflammatory response and oxidative stress, and decrease autophagy of hippocampal neurons in CFS rats by regulating the PI3K/Beclin1 signaling pathway.
3.Herbal Cake-separated Moxibustion Improves Cognitive Function in Rat Model of Chronic Fatigue Syndrome via PI3K/Beclin1 Signaling Pathway
Chuntao ZHAI ; Yawei HOU ; Linjuan SHI ; Yixiao WANG ; Yuefeng TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):149-158
ObjectiveTo explore the mechanism of moxibustion with the classical prescription Xiaoyaosan as the herbal cake in improving cognitive function in the rat model of chronic fatigue syndrome (CFS) by regulating autophagy via phosphatidylinosiol 3-kinase (PI3K)/autophagy key molecule yeast Atg6 homologue 1 (Beclin1) signaling pathway. MethodsFifty SPF-grade SD rats aged 6-8 weeks were randomly grouped as follows: control, model, sham cake-separated moxibustion, intragastric administration with Chinese herbs, and herbal cake-separated moxibustion, each with 10 rats. The other groups except the control group were subjected to modeling by exhaustive swimming and chronic restraint stress for 21 days, and the behavioral test was performed after modeling. Herbal cake-separated moxibustion and sham cake-separated moxibustion were carried out at Shenque (CV8), Guanyuan (CV4), Zusanli (ST36), and Qimen (LR14). The rats in the intragastric administration with Chinese herbs group were treated with Xiaoyaosan suspension by gavage. After continuous intervention for 10 days, rat behaviors were observed and the levels of interleukin (IL)-4, IL-10, blood urine nitrogen (BUN), and malondiadehyde (MDA) in the serum and hippocampal tissue of rats in each group were determined by enzyme-linked immunosorbent assay. The morphological changes of the hippocampal tissue were observed by hematoxylin-eosin staining. The relative mRNA and protein levels of PI3K and Beclin1 in the hippocampal tissue were determined by Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. ResultsCompared with the normal group, the model group showed reduced crossings, shortened residence time in the target quadrant, and prolonged average escape latency on days 1, 2, 3, and 4 (P<0.01) in the Morris water maze test. In the new object recognition test, the model group showed decreased recognition index of exploration time and recognition index of exploration times (P<0.01), lowered levels of IL-4 and IL-10 and elevated levels of BUN and MDA in the serum and hippocampus (P<0.01), and down-regulated mRNA and protein levels of PI3K and Beclin1 (P<0.01). After treatment, compared with the model group, each treatment group showed increased crossings, prolonged residence time in the target quadrant, and shortened average escape latency on days 1, 2, 3, and 4 (P<0.05, P<0.01). In addition, the treatment elevated the levels of IL-4 and IL-10 and lowered the levels of BUN and MDA in the serum and hippocampus (P<0.05, P<0.01), and up-regulated the mRNA and protein levels of PI3K and Beclin1 (P<0.05, P<0.01). Compared with the sham cake-separated moxibustion group, the herbal cake-separated moxibustion group and the intragastric administration with Chinese herbs group showed increased crossings, prolonged residence time in the target quadrant (P<0.05), elevated level of IL-4, lowered level of MDA, and up-regulated relative mRNA and protein levels of PI3K and Beclin1 in the hippocampus (P<0.05, P<0.01). The serum levels of MDA and BUN in the intragastric administration with Chinese herbs group were significantly decreased. The average escape latency of the herbal cake-separated moxibustion group was significantly shortened on days 2 and 3 (P<0.05, P<0.01). In addition, this group showed elevated serum level of IL-4, lowered serum level of MDA, and declined BUN level in the hippocampus (P<0.05, P<0.01). Compared with the intragastric administration with Chinese herbs group, the herbal cake-separated moxibustion group presented risen serum level of IL-4 and up-regulated mRNA level of Beclin1 (P<0.05) and relative protein levels of PI3K and Beclin1 (P<0.05, P<0.01). ConclusionHerbal cake-separated moxibustion can effectively alleviate fatigue and improve the memory, reduce inflammatory response and oxidative stress, and decrease autophagy of hippocampal neurons in CFS rats by regulating the PI3K/Beclin1 signaling pathway.
4.Mechanism of immunosuppression in rats by herb-partitioned moxibustion based on transcriptome sequencing technology
Yuefeng TIAN ; Luojie XIONG ; Huifang WANG ; Chuntao ZHAI ; Wei LI
Chinese Journal of Tissue Engineering Research 2025;29(5):978-988
BACKGROUND:Immunosuppression leads to impaired body immune function and aggravates the disease.Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body,but its regulatory mechanism has not been elucidated. OBJECTIVE:To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity. METHODS:Twenty-four Sprague-Dawley rats were randomly assigned to three groups:control,model,and herb-partitioned moxibustion groups,with eight rats in each group.The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days.No interventions were administered to the control and model groups after modeling.In contrast,the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan,Shenque,Guanyuan,and Zusanli acupoints using a combination of moxa and herbal cakes,once a day,for 10 consecutive days,with samples being collected the day after the end of the intervention.Peripheral blood was collected from all groups of rats to measure their white blood cell count.RNA-seq was performed on the Illumina sequencing platform,and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases. RESULTS AND CONCLUSION:Compared with the control group,the model group exhibited a significant decrease in white blood cell count(P<0.001).RNA-seq analysis identified 3 026 differentially expressed genes between the model and control groups,with 1 565 upregulated and 1 461 downregulated.There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group,with 280 upregulated and 255 downregulated.The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group.However,after moxibustion with herbal cakes,these genes were upregulated.Protein-protein interaction network analysis identified 10 core targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,Mx1,Syk,Hspa1a,and Ret.According to GO and KEGG analyses,moxibustion with herbal cakes regulated the body through pathways related to immune response,viruses,angiogenesis,and the autoimmune system.To conclude,there is a significant association between herbal cake-separated moxibustion intervention and immune suppression targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,and Mx1.The intervention exhibits regulatory effects in the pathways related to immune responses,viral activities,and angiogenesis.
5.Active surveillance of drug safety and influential factors in pediatric patients with Mycoplasma pneumoniae pneumonia
Yuefeng WANG ; Qiang GAO ; Ke QIN
China Pharmacy 2025;36(22):2838-2844
OBJECTIVE To analyze adverse drug reactions (ADRs) in pediatric patients with Mycoplasma pneumoniae pneumonia (MPP), and investigate the influential factors associated with the occurrence of ADRs. METHODS Clinical records of pediatric patients with MPP in Xinxiang Central Hospital (hereinafter referred to as “our hospital”) from Oct. 1st, 2023 to Sept. 30th, 2024 collected from the Hospital Information System were retrospectively reviewed using the global trigger tool (GTT). The positive triggers were further reviewed to confirm the occurrence of ADRs, and the characteristics of those ADRs were analyzed. The children were divided into the ADR group and the non-ADR group. Univariate analysis and multivariate Logistic regression analysis were performed to investigate the potential influential factors for ADRs in MPP children. RESULTS A total of 1 325 MPP children were included. Out of 24 designed triggers, 14 were positive, with a positive trigger rate of 58.3%. There were 1 301 positive trigger cases, involving 846 pediatric patients. After assessment, 399 cases of ADRs were identified in 372 pediatric patients, yielding a positive predictive value of triggers at 30.7%. The main types of ADRs were abnormal blood cells (230 cases, 57.6%), skin and its appendages damage (78 cases, 19.5%), and gastrointestinal system damage (42 cases, 10.5%), which predominantly occurred 4 days or more, within 2-3 days and within 2-3 days after medication, respectively. They were more prevalent in pediatric patients aged>3-10 years (295 cases, 79.3%). The major categories of drugs associated with ADRs included intravenous corticosteroids (209 cases), cephalosporins (180 cases), macrolides (38 cases), tetracyclines (28 cases) and traditional Chinese medicine (TCM) injections (28 cases). These ADR reports were classified into 3 types: 1 serious case, 394 general cases and 4 new cases. In terms of causality assessment, 1 case was assessed as certain, 19 cases as probable, and 379 cases as possible. In terms of ADR outcomes, 151 cases fully recovered, while the outcomes for 248 cases remained unknown. Univariate analysis and multivariate Logistic regression analysis revealed that a history of allergies [OR=5.231, 95%CI (2.004, 13.656), P<0.05], the number of medications used [OR=1.064, 95%CI (1.027, 1.103), P<0.05], and intravenous corticosteroid administration [OR=3.223, 95%CI (2.341, 4.437), P<0.05] were independent risk factors of ADRs in pediatric patients with MPP. The use of azithromycin [OR=0.544, 95%CI (0.376, 0.786), P<0.05] was identified as an independent protective factor against the occurrence of ADRs in pediatric patients with MPP. CONCLUSIONS Children with MPP in our hospital exhibit a higher incidence of ADRs, primarily involving abnormal blood cells, skin and its appendages damage, and gastrointestinal system damage. A history of allergies, the number of medications used, intravenous corticosteroid administration, and the use of azithromycin are associated with the occurrence of ADR in MPP pediatric patients.
6.Birth weight, ideal cardiovascular health metrics in adulthood, and incident cardiovascular disease
Ying SUN ; Bin WANG ; Yuefeng YU ; Yuying WANG ; Xiao TAN ; Jihui ZHANG ; Lu QI ; Yingli LU ; Ningjian WANG
Chinese Medical Journal 2024;137(10):1160-1168
Background::Prenatal and postnatal factors may have joint effects on cardiovascular health, and we aimed to assess the joint association of birth weight and ideal cardiovascular health metrics (ICVHMs) prospectively in adulthood with incident cardiovascular disease (CVD).Methods::In the UK Biobank, 227,833 participants with data on ICVHM components and birth weight and without CVD at baseline were included. The ICVHMs included smoking, body mass index, physical activity, diet information, total cholesterol, blood pressure, and hemoglobin A1c. The Cox proportional hazards model was used to estimate the hazard ratios (HRs) and 95% confidence intervals (CIs) in men and women.Results::Over a median follow-up period of 13.0 years (2,831,236 person-years), we documented 17,477 patients with incident CVD. Compared with participants with birth weights of 2.5-4.0 kg, the HRs (95% CIs) of CVD among those with low birth weights was 1.08 (1.00-1.16) in men and 1.23 (1.16-1.31) in women. The association between having a birth weight <2.5 kg and CVD risk in men was more prominent for those aged <50 years than for those of older age ( P for interaction = 0.026). Lower birth weight and non-ideal cardiovascular health metrics were jointly related to an increased risk of CVD. Participants with birth weights <2.5 kg and ICVHMs score 0-1 had the highest risk of incident CVD (HR [95% CI]: 3.93 [3.01-5.13] in men; 4.24 [3.33-5.40] in women). The joint effect (HR [95% CI]: 1.36 [1.17-1.58]) could be decomposed into 24.7% (95% CI: 15.0%-34.4%) for a lower birth weight, 64.7% (95% CI: 56.7%-72.6%) for a lower ICVHM score, and 10.6% (95% CI: 2.7%-18.6%) for their additive interaction in women. Conclusions::Birth weight and ICVHMs were jointly related to CVD risk. Attaining a normal birth weight and ideal ICVHMs may reduce the risk of CVD, and a simultaneous improvement of both prenatal and postnatal factors could further prevent additional cases in women.
7.Life’s Essential 8 and risk of non-communicable chronic diseases: Outcome-wide analyses
Yuetian YU ; Ying SUN ; Yuefeng YU ; Yuying WANG ; Chi CHEN ; Xiao TAN ; Yingli LU ; Ningjian WANG
Chinese Medical Journal 2024;137(13):1553-1562
Background::Life’s Simple 7, the former construct of cardiovascular health (CVH) has been used to evaluate adverse non-communicable chronic diseases (NCDs). However, some flaws have been recognized in recent years and Life’s Essential 8 has been established. In this study, we aimed to analyze the association between CVH defined by Life’s Essential 8 and risk of 44 common NCDs and further estimate the population attributable fractions (PAFs) of low-moderate CVH scores in the 44 NCDs.Methods::In the UK Biobank, 170,726 participants free of 44 common NCDs at baseline were included. The Life’s Essential 8 composite measure consists of four health behaviours (diet, physical activity, nicotine exposure, and sleep) and four health factors (body mass index, non-high density lipoprotein cholesterol, blood glucose, and blood pressure), and the maximum CVH score was 100 points. CVH score was categorized into low, moderate, and high groups. Participants were followed up for 44 NCDs diagnosis across 10 human system disorders according to the International Classification of Diseases 10th edition (ICD-10) code using linkage to national health records until 2022. Cox proportional hazard models were used in this study. The hazard ratios (HRs) and PAFs of 44 NCDs associated with CVH score were examined.Results::During the median follow-up of 10.85 years, 58, 889 incident NCD cases were documented. Significant linear dose-response associations were found between higher CVH score and lower risk of 25 (56.8%) of 44 NCDs. Low-moderate CVH (<80 points) score accounted for the largest proportion of incident cases in diabetes (PAF: 80.3%), followed by gout (59.6%), sleep disorder (55.6%), chronic liver disease (45.9%), chronic kidney disease (40.9%), ischemic heart disease (40.8%), chronic obstructive pulmonary disease (40.0%), endometrium cancer (35.8%), lung cancer (34.0%), and heart failure (34.0%) as the top 10. Among the eight modifiable factors, overweight/obesity explained the largest number of cases of incident NCDs in endocrine, nutritional, and metabolic diseases (35.4%), digestive system disorders (21.4%), mental and behavioral disorders (12.6%), and cancer (10.3%); however, the PAF of ideal sleep duration ranked first in nervous system (27.5%) and neuropsychiatric disorders (9.9%).Conclusions::Improving CVH score based on Life’s Essential 8 may lower risk of 25 common NCDs. Among CVH metrics, avoiding overweight/obesity may be especially important to prevent new cases of metabolic diseases, NCDs in digestive system, mental and behavioral disorders, and cancer.
8.Establishment of a rapid detection method for carbapenem and quinolone resistant nucleic acid colloidal gold test strips and development of a reagent kit
Beizhen PAN ; Jifei YANG ; Yuefeng WANG ; Yan LIU ; Yujiao ZHOU ; Yuhe MA ; Liyuan SUN
Chinese Journal of Immunology 2024;40(11):2386-2390,2398
Objective:To establish a method for rapid detection of OXA and par C resistance genes of Acinetobacter baumannii(Ab)by double nucleic acid colloidal gold strip and to develop kit.Methods:DNA of Ab was extracted by heating and boiling method.OXA and par C genes sequences of Ab were selected as target gene fragments based on NCBI.Primers were designed and labeled with 6-FAM,digoxin and biotin,respectively.Drug resistance gene detection reagents were developed,and nucleic acid gold test strips were used for rapid and visual detection.Molecular cloning and sequencing techniques were used to clone positive control samples and evaluate specificity,sensitivity and stability of kit.Results:DNA concentration and purity of Ab extracted by boiling method were good.Homology between cloned and sequenced plasmid DNA and gene sequence in GenBank database was 100%,respectively.Speci-ficity of kit was good,with only Ab showing positive results and other bacterial genera showing negative results;DNA concentration of Ab in double nucleic acid colloidal gold test strip decreased to 10-3 ng/μl,a red line still appeared,whose sensitivity was 10 times higher consistent with minimum detection limit of electrophoresis 10-2 ng/μl;test kits were tested at 3rd,6th and 9th months,and showed good stability.Conclusion:Double resistance detection kit established in this study can simultaneously detect OXA and par C resis-tance of Ab,who has advantages of high sensitivity,strong specificity,rapid and simple,and provides a new method for detection of carbapenem and quinolone antibiotic resistance of Ab.
9.Diversity of the T Cell Receptor β Chain Complementarity-Determining Region 3 in Peripheral Blood of Typical Syndromes Cirrhosis of the Liver:An Analysis Based on Immune Repertoire Sequencing
Jia WANG ; Yuting HU ; Yuefeng GONG ; Jian ZHU ; Shibing SU ; Yiyu LU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(7):1916-1924
Objective In this study,the diversity of the TCR β chain CDR3 in peripheral blood of patients with different typical Traditional Chinese Medicine(TCM)syndromes of liver cirrhosis was analyzed by immune repertoire sequencing,the material basis and regularity of the syndromes of liver cirrhosis was discussed.Methods 20 patients with cirrhosis,including liver and gallbladder damp heat(LGHD),liver depression and spleen deficiency(LDSD),and liver and kidney yin deficiency(LKYD)were enrolled into case group,and 10 healthy patients were used as the healthy control group.DNA was extracted from peripheral blood samples,and multiplex PCR amplification of TCR β chain CDR3 was performed,followed by high-throughput sequencing of the products to analyze the diversity of the TCR β chain CDR3.Results The nt sequence numbers unique to CDR3 and aa sequence numbers unique to CDR3 of LDSD between liver cirrhosis syndromes were less than LKYD(P<0.05).Clonality,Pielous,Shannon.Index and DE50 of LGHD and LKYD had significant differences(P<0.05)between two groups,as well as the frequency of multiple fragments in V and J regions and V-J gene recombination of LGHD vs.LDSD,LGHD vs.LKYD,and LKYD vs.LDSD,respectively(P<0.05).LGHD vs.LDSD,TRBV21-1,TRBV12-4,TRBV11-1 subtype and 7 pairs of V-J recombination have statistical differences(P<0.05).LGHD vs.LKYD,TRBV10-2,TRBV7-6,TRBV5-8 subtypes and 30 pairs of V-J recombination were statistically different(P<0.05).LDSD vs.LKYD,there were statistical differences between TRBJ1-5 subtype and 18 pairs of V-J recombination(P<0.05).Conclusions The present study was conducted to find that the diversity of TCR CDR3 in liver cirrhosis syndrome is significantly different and conforms to the regularity of syndrome from excess to deficiency by explore the immunological characteristics of different TCM syndromes of liver cirrhosis,and to provide new support for the objective basis of"combination of disease and TCM syndrome"and"diagnosis and treatment".We explored the different expression patterns and specificity of adaptive immune gene rearrangement in patients with different TCM syndromes to identify different expression patterns and specific markers of adaptive immune gene rearrangements of typical TCM syndromes in liver cirrhosis.
10.Expression of Serum miR-138-5p and miR-212-5p Levels and Clinical Prognosis Value in Prostate Cancer
Long ZHANG ; Yuefeng LI ; Tao WU ; Linjie ZHU ; Haidong WANG
Journal of Modern Laboratory Medicine 2024;39(5):30-34
Objective To investigate the clinical prognosis value of serum microRNA(miR)-138-5p and miR-212-5p levels in prostate cancer.Methods A total of 95 cases of prostate cancer patients admitted to the First Hospital of Handan from July 2020 to June 2021 were collected.Based on follow-up records for two years after surgery,these patients were separated into a poor prognosis group(n=52)and a good prognosis group(n=43),and 48 healthy volunteers who underwent a physical examination at the hospital were collected as the healthy control group.In addition,quantitative real-time fluorescence-PCR(qRT-PCR)was applied to detect the relative expression levels of miR-138-5p and miR-212-5p in the serum of study subjects,and the clinical data of the patients were collected and analyzed.Multivariate COX regression was applied to analyze factors that affected the prognosis of prostate cancer patients.The predictive value of serum miR-138-5p and miR-212-5p for the prognosis of prostate patients was tested,and receiver operating characteristic(ROC)curves were plotted.Pearson method was applied to analyze the correlation between the expression of serum miR-138-5p and miR-212-5p and the Gleason score.Results Compared to the healthy control group,the serum levels of miR-138-5p(0.88±0.10,0.83±0.09 vs 1.01±0.10),and miR-212-5p(0.75±0.09,0.71±0.08 vs 1.02±0.11)were lower in the good prostate cancer prognosis group and poor prostate cancer prognosis group,and the differences were significant(t=14.021,22.275;9.825,18.063,all P<0.05).The prognosis of prostate cancer patients was related to TNM staging,bone metastasis,tissue differentiation degree,preoperative PSA level,and Gleason score(x2=4.417~7.187,t=14.235,all P<0.05).Serum miR-138-5p[HR(95%CI):0.871(0.785~0.966)],and miR-212-5p[HR(95%CI):0.822(0.725~0.932)]were protective factors for poor prognosis(all P<0.05).While Gleason score[HR(95%CI):1.253(1.026~1.530)],TNM stage[HR(95%CI):1.224(1.024~1.463)],bone metastasis[HR(95%CI):1.398(1.036~1.887)],tissue differentiation degree[HR(95%CI):1.520(1.146~2.016)]and PSA level[HR(95%CI):1.426(1.094~1.858)]were all risk factors for poor prognosis(all P<0.05).The AUC(95%CI)of serum miR-138-5p and miR-212-5p were 0.883(95%CI:0.801~0.940)and 0.863(95%CI:0.777~0.925),respectively.Serum miR-138-5p and miR-212-5p were negatively correlated with the Gleason score(r=-0.610,-0.420,all P<0.05).Conclusion Serum miR-138-5p and miR-212-5p levels are elevated in patients with poor prostate cancer prognosis and may have a certain auxiliary predictive value for prostate cancer patient prognosis.

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