Objective:To explore the abnormal expression and significance of circ-CBLB/miR-486-5p in patients with rheumatoid arthritis of spleen deficiency and dampness excess type.Methods:The 30 healthy individuals included in ...Objective:To explore the abnormal expression and significance of circ-CBLB/miR-486-5p in patients with rheumatoid arthritis of spleen deficiency and dampness excess type.Methods:The 30 healthy individuals included in the method were from the Physical Examination Center of Anhui Provincial Hospital of Traditional Chinese Medicine,and the 60 rheumatoid arthritis patients included were from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine.The disease activity score of patients with rheumatoid arthritis was evaluated using VAS score and DAS28 score,joint symptoms and spleen deficiency syndrome score were evaluated using graded quantification method,immune inflammation indicators were detected using relevant instruments,inflammatory cytokines were detected using ELISA method,macrophage markers were detected using FCM method,and pathway gene expression was detected using RT-qPCR;Evaluate the predictive effect of circ-CBLB and miR-486-5p on disease activity in rheumatoid arthritis using ROC curves.Results:(1)miR-486-5p,CD14^(+)CD86^(+),ESR,CRP,RF,Anti CCP Ab,IL-6,TNF in patients with rheumatoid arthritis-αThe levels of circ-CBLB,CD14^(+)CD163^(+),IL-4,and IL-10 were significantly higher than those of healthy individuals;(2)The expression level of circ-CBLB in patients with rheumatoid arthritis is positively correlated with CD14^(+)CD163^(+),and negatively correlated with miR-486-5p and CD14^(+)CD86^(+);The expression level of miR-486-5p is negatively correlated with CD14^(+)CD163^(+)and positively correlated with CD14^(+)CD86^(+);There is a negative correlation between CD14^(+)CD86^(+)and CD14^(+)CD163^(+);ESR is negatively correlated with circ-CBLB,and positively correlated with miR-486-5p,CD14^(+)CD86^(+),CRP;CRP is negatively correlated with circ-CBLB,CD14^(+)CD163^(+),and positively correlated with CD14^(+)CD86^(+),ESR;(3)The expression level of circ-CBLB in patients with rheumatoid arthritis is negatively correlated with joint tenderness,morning stiffness,lack of qi and lazy speech,and postprandial abdominal distension score;The expression level of miR-486-5p is positively correlated with the scores of joint tenderness and decreased appetite.(4)The ROC curve shows that in terms of circ-CBLB,ESR,CRP,VAS,and DAS28 AUC are 0.662(P=0.032),0.658(P=0.035),0.516(P=0.830),and 0.791(P=0.000),respectively.In terms of miR-486-5p,ESR,CRP,VAS,and DAS28 AUC were 0.566(P=0.385),0.511(P=0.883),0.592(P=0.223),and 0.727(P=0.003),respectively.Conclusion:The abnormal expression of circ CBLB and miR-486-5p in peripheral blood mononuclear cell of patients with rheumatoid arthritis of spleen deficiency and dampness excess type is related to inflammatory polarization markers,immune inflammation,disease activity,joint symptoms and spleen deficiency syndrome of rheumatoid arthritis,and the low expression of circ CBLB and high expression of miR-486-5p have certain predictive value for disease activity of rheumatoid arthritis.展开更多
Objective: to explore the mechanism of transportation and transformation of dampness by the way of the expression of organic anion transporting polypeptide (oatp) superfamily member 2a1 (oatp2a1) mRNA in rat with sple...Objective: to explore the mechanism of transportation and transformation of dampness by the way of the expression of organic anion transporting polypeptide (oatp) superfamily member 2a1 (oatp2a1) mRNA in rat with spleen deficiency syndrome and the significance in transportation and transformation of dampness. Methods: 32 wistar male rats were divided randomly into four groups: normal group (n = 6), normal + AA group (n = 6), spleen deficiency group (n = 10), Spleen deficiency + AA group (n = 10). After reserpine-induced spleen deficiency model was made, intragastric administration of aristolochic acid (AA) was adopted for three days, the expression of oatp2a1 mRNA were detected in the tissues of lung, liver, kidney, stomach, small intestine and large intestine in four groups by using Fluorescent Quantitative-Polymerase Chain Reaction (FQ-PCR). Results: the expression of oatp2a1 mRNA in above six tissues could be detected. The ex-pression of oatp2a1 mRNA in liver tissue of rat with spleen deficiency syndrome was up-regulated compared to normal group (P = 0.035, P < 0.05), the expression of oatp2a1 mRNA in small intestinal tissue of rat with spleen deficiency syndrome was down-regulated compared to normal group (P = 0.004, P < 0.01), the expression of oatp2a1 in intestinal tissue in normal + AA group is down-regulated compared to normal group (P = 0.032, P < 0.05). Conclusions: oatp2a1 might be one of the material basis involved in transportation and transformation of dampness. The changes of expression of oatp2a1 mRNA in small intestine, liver tissue suggests that small intestine, liver might play an important role in the transportation and transformation of dampness in the state of spleen deficiency. We further concluded that the function of spleen’s governing transportation and transformation of dampness was not only including the function of the gastrointestinal, but also part of the liver function in some degree, which needs to be further studied.展开更多
Objective:This study was designed to explore the characteristics of gut microbiota in CHB patients with two most common traditional Chinese medicine(TCM)Syndromes—damp heat syndrome and liver depression and spleen de...Objective:This study was designed to explore the characteristics of gut microbiota in CHB patients with two most common traditional Chinese medicine(TCM)Syndromes—damp heat syndrome and liver depression and spleen deficiency syndrome.Methods:According to the diagnostic criteria of Western medicine,TCM and screening exclusion criteria,65 cases of CHB with damp heat syndrome and 28 cases of CHB with liver depression and spleen deficiency syndrome were finally included in the study.All the basic information was gathered and the fresh fecal samples were collected for 16S rDNA sequencing.16S rDNA of gut microbiota was sequenced using Illumina hiseq 2,500 high-throughput sequencing platform.Based on the optimized sequence,Operational Taxonomic Units(OTU)clustering analysis and taxonomic annotation were carried out.Results:The difference in relative abundance of gut microbiota was significant between damp heat syndrome and liver depression and spleen deficiency syndrome in CHB patients.Cyanobacteria was only found in damp heat syndrome.The relative abundance of Erysipelotrichia and Subdoligranulum were higher in liver depression and spleen deficiency syndrome,while the relative abundance of Rhodospirillales,Alphaproteobacteria and Lachnospira were higher in the damp heat syndrome.LDA Effect Size(LEfSe)analysis showed that Lachnospira,Olsenella and Subdoligranulum had significant difference in species among the two TCM syndromes.Conclusion:The different characteristics of gut microbiota in the two TCM syndromes of CHB patients may play an important role in syndrome formation of TCM,which provides a new field of vision for the accurate diagnosis and treatment of TCM.展开更多
Poria is an important medicine for inducing diuresis to drain dampness from the middle energizer.However,the specific effective components and the potential mechanism of Poria remain largely unknown.To identify the ef...Poria is an important medicine for inducing diuresis to drain dampness from the middle energizer.However,the specific effective components and the potential mechanism of Poria remain largely unknown.To identify the effective components and the mechanism of Poria water extract(PWE)to treat dampness stagnancy due to spleen deficiency syndrome(DSSD),a rat model of DSSD was established through weight-loaded forced swimming,intragastric ice-water stimulation,humid living environment,and alternate-day fasting for 21 days.After 14 days of treatment with PWE,the results indicated that PWE increased fecal moisture percentage,urine output,D-xylose level and weight;amylase,albumin,and total protein levels;and the swimming time of rats with DSSD to different extents.Eleven highly related components were screened out using the spectrum-effect relationship and LC-MS.Mechanistic studies revealed that PWE significantly increased the expression of serum motilin(MTL),gastrin(GAS),ADCY5/6,p-PKAα/β/γcat,and phosphorylated cAMP-response element binding protein in the stomach,and AQP3 expression in the colon.Moreover,it decreased the levels of serum ADH,the expression of AQP3 and AQP4 in the stomach,AQP1 and AQP3 in the duodenum,and AQP4 in the colon.PWE induced diuresis to drain dampness in rats with DSSD.Eleven main effective components were identified in PWE.They exerted therapeutic effect by regulating the AC-cAMP-AQP signaling pathway in the stomach,MTL and GAS levels in the serum,AQP1 and AQP3 expression in the duodenum,and AQP3 and AQP4 expression in the colon.展开更多
基金the Seventh Batch of Inheritance Project of Academic Experience of National Veteran TCM Experts of the National Administration of Traditional Chinese Medicine (GZYYRJH[2022]No. 76)National Natural Science Foundation General Project (No.81973655)+4 种基金National Natural Science Foundation General Project (No.82274501)Anhui Provincial Excellent Youth Talent Support Program for Universities (Anhui Education Secretary [2022]No. 11)Key Laboratory of Xin'an Medical Education Department (No.2020xayx04)Natural Science Research Project of Universities in Anhui Province (No.KJ2020A0397)Anhui NewEra Education Quality Engineering Project (No.2022xscx103)。
文摘Objective:To explore the abnormal expression and significance of circ-CBLB/miR-486-5p in patients with rheumatoid arthritis of spleen deficiency and dampness excess type.Methods:The 30 healthy individuals included in the method were from the Physical Examination Center of Anhui Provincial Hospital of Traditional Chinese Medicine,and the 60 rheumatoid arthritis patients included were from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine.The disease activity score of patients with rheumatoid arthritis was evaluated using VAS score and DAS28 score,joint symptoms and spleen deficiency syndrome score were evaluated using graded quantification method,immune inflammation indicators were detected using relevant instruments,inflammatory cytokines were detected using ELISA method,macrophage markers were detected using FCM method,and pathway gene expression was detected using RT-qPCR;Evaluate the predictive effect of circ-CBLB and miR-486-5p on disease activity in rheumatoid arthritis using ROC curves.Results:(1)miR-486-5p,CD14^(+)CD86^(+),ESR,CRP,RF,Anti CCP Ab,IL-6,TNF in patients with rheumatoid arthritis-αThe levels of circ-CBLB,CD14^(+)CD163^(+),IL-4,and IL-10 were significantly higher than those of healthy individuals;(2)The expression level of circ-CBLB in patients with rheumatoid arthritis is positively correlated with CD14^(+)CD163^(+),and negatively correlated with miR-486-5p and CD14^(+)CD86^(+);The expression level of miR-486-5p is negatively correlated with CD14^(+)CD163^(+)and positively correlated with CD14^(+)CD86^(+);There is a negative correlation between CD14^(+)CD86^(+)and CD14^(+)CD163^(+);ESR is negatively correlated with circ-CBLB,and positively correlated with miR-486-5p,CD14^(+)CD86^(+),CRP;CRP is negatively correlated with circ-CBLB,CD14^(+)CD163^(+),and positively correlated with CD14^(+)CD86^(+),ESR;(3)The expression level of circ-CBLB in patients with rheumatoid arthritis is negatively correlated with joint tenderness,morning stiffness,lack of qi and lazy speech,and postprandial abdominal distension score;The expression level of miR-486-5p is positively correlated with the scores of joint tenderness and decreased appetite.(4)The ROC curve shows that in terms of circ-CBLB,ESR,CRP,VAS,and DAS28 AUC are 0.662(P=0.032),0.658(P=0.035),0.516(P=0.830),and 0.791(P=0.000),respectively.In terms of miR-486-5p,ESR,CRP,VAS,and DAS28 AUC were 0.566(P=0.385),0.511(P=0.883),0.592(P=0.223),and 0.727(P=0.003),respectively.Conclusion:The abnormal expression of circ CBLB and miR-486-5p in peripheral blood mononuclear cell of patients with rheumatoid arthritis of spleen deficiency and dampness excess type is related to inflammatory polarization markers,immune inflammation,disease activity,joint symptoms and spleen deficiency syndrome of rheumatoid arthritis,and the low expression of circ CBLB and high expression of miR-486-5p have certain predictive value for disease activity of rheumatoid arthritis.
文摘Objective: to explore the mechanism of transportation and transformation of dampness by the way of the expression of organic anion transporting polypeptide (oatp) superfamily member 2a1 (oatp2a1) mRNA in rat with spleen deficiency syndrome and the significance in transportation and transformation of dampness. Methods: 32 wistar male rats were divided randomly into four groups: normal group (n = 6), normal + AA group (n = 6), spleen deficiency group (n = 10), Spleen deficiency + AA group (n = 10). After reserpine-induced spleen deficiency model was made, intragastric administration of aristolochic acid (AA) was adopted for three days, the expression of oatp2a1 mRNA were detected in the tissues of lung, liver, kidney, stomach, small intestine and large intestine in four groups by using Fluorescent Quantitative-Polymerase Chain Reaction (FQ-PCR). Results: the expression of oatp2a1 mRNA in above six tissues could be detected. The ex-pression of oatp2a1 mRNA in liver tissue of rat with spleen deficiency syndrome was up-regulated compared to normal group (P = 0.035, P < 0.05), the expression of oatp2a1 mRNA in small intestinal tissue of rat with spleen deficiency syndrome was down-regulated compared to normal group (P = 0.004, P < 0.01), the expression of oatp2a1 in intestinal tissue in normal + AA group is down-regulated compared to normal group (P = 0.032, P < 0.05). Conclusions: oatp2a1 might be one of the material basis involved in transportation and transformation of dampness. The changes of expression of oatp2a1 mRNA in small intestine, liver tissue suggests that small intestine, liver might play an important role in the transportation and transformation of dampness in the state of spleen deficiency. We further concluded that the function of spleen’s governing transportation and transformation of dampness was not only including the function of the gastrointestinal, but also part of the liver function in some degree, which needs to be further studied.
基金The Sixth Group of National Senior Chinese Medicine Experts’ Academic Experience Inheritance Project [approval:State Administration of Traditional Chinese Medicine (2017) No.29]the Foshan High Level Medical Key Discipline Construction Project and "Peak Climbing Plan" for Foshan High Level Hospital Construction[approval:Foshan Municipal Government Office,No. 2019 (01)]
文摘Objective:This study was designed to explore the characteristics of gut microbiota in CHB patients with two most common traditional Chinese medicine(TCM)Syndromes—damp heat syndrome and liver depression and spleen deficiency syndrome.Methods:According to the diagnostic criteria of Western medicine,TCM and screening exclusion criteria,65 cases of CHB with damp heat syndrome and 28 cases of CHB with liver depression and spleen deficiency syndrome were finally included in the study.All the basic information was gathered and the fresh fecal samples were collected for 16S rDNA sequencing.16S rDNA of gut microbiota was sequenced using Illumina hiseq 2,500 high-throughput sequencing platform.Based on the optimized sequence,Operational Taxonomic Units(OTU)clustering analysis and taxonomic annotation were carried out.Results:The difference in relative abundance of gut microbiota was significant between damp heat syndrome and liver depression and spleen deficiency syndrome in CHB patients.Cyanobacteria was only found in damp heat syndrome.The relative abundance of Erysipelotrichia and Subdoligranulum were higher in liver depression and spleen deficiency syndrome,while the relative abundance of Rhodospirillales,Alphaproteobacteria and Lachnospira were higher in the damp heat syndrome.LDA Effect Size(LEfSe)analysis showed that Lachnospira,Olsenella and Subdoligranulum had significant difference in species among the two TCM syndromes.Conclusion:The different characteristics of gut microbiota in the two TCM syndromes of CHB patients may play an important role in syndrome formation of TCM,which provides a new field of vision for the accurate diagnosis and treatment of TCM.
基金supported by the Key Technologies Research and Development Program(No.2017YFC1703004)Hubei Technological Innovation Special Fund(No.2019ACA121).
文摘Poria is an important medicine for inducing diuresis to drain dampness from the middle energizer.However,the specific effective components and the potential mechanism of Poria remain largely unknown.To identify the effective components and the mechanism of Poria water extract(PWE)to treat dampness stagnancy due to spleen deficiency syndrome(DSSD),a rat model of DSSD was established through weight-loaded forced swimming,intragastric ice-water stimulation,humid living environment,and alternate-day fasting for 21 days.After 14 days of treatment with PWE,the results indicated that PWE increased fecal moisture percentage,urine output,D-xylose level and weight;amylase,albumin,and total protein levels;and the swimming time of rats with DSSD to different extents.Eleven highly related components were screened out using the spectrum-effect relationship and LC-MS.Mechanistic studies revealed that PWE significantly increased the expression of serum motilin(MTL),gastrin(GAS),ADCY5/6,p-PKAα/β/γcat,and phosphorylated cAMP-response element binding protein in the stomach,and AQP3 expression in the colon.Moreover,it decreased the levels of serum ADH,the expression of AQP3 and AQP4 in the stomach,AQP1 and AQP3 in the duodenum,and AQP4 in the colon.PWE induced diuresis to drain dampness in rats with DSSD.Eleven main effective components were identified in PWE.They exerted therapeutic effect by regulating the AC-cAMP-AQP signaling pathway in the stomach,MTL and GAS levels in the serum,AQP1 and AQP3 expression in the duodenum,and AQP3 and AQP4 expression in the colon.
