1.Comparative Study on the Effects of Qingchang Wenzhong Decoction (清肠温中方) on the Intestinal Mucus Barrier in Conventional and Pseudo-Germ-Free Model Mice of Acute Ulcerative Colitis
Mingxu GAO ; Leilei LIU ; Lijie LU ; Jiali WANG ; Juncong HU ; Yangzhe LIN ; Qirui LIU ; Yue DONG ; Luyue WANG ; Zeyu XUE ; Junxiang LI ; Lei SHI
Journal of Traditional Chinese Medicine 2026;67(14):1528-1537
ObjectiveTo explore the mechanisms of Qingchang Wenzhong Decoction (清肠温中方, QWD) in treating ulcerative colitis (UC). MethodsA total of 100 male C57BL/6J mice were assigned to a pseudo-germ-free group (n=45) and a conventional group (n=55) using a random number table. Pseudo-germ-free model was established by giving antibiotic treatment and subsequently subjected to acute UC induction. After the model was successfully established, the 45 pseudo-germ-free acute UC mice were randomly allocated to a pseudo-germ-free model group, a pseudo-germ-free QWD group, and a pseudo-germ-free mesalazine group, with 15 mice in each group. The 55 conventional mice were randomly divided into a normal control group (n=10), as well as a conventional model group, a conventional QWD group, and a conventional mesalazine group, with 15 mice in each group. Except for the normal control group, acute UC models were established in all other groups. Mice in the control group received no intervention. From day 32 of the experiment, mice in the pseudo-germ-free QWD group and the conventional QWD group were administered with QWD granules at a dose of 9.71 g/(kg·d) by gavage with deionized water, while those in the pseudo-germ-free mesalazine group and the conventional mesalazine group were given mesalazine enteric-coated tablets at a dose of 151.67 mg/(kg·d) by gavage. Mice in the remaining groups received an equal volume of deionized water once daily for 7 consecutive days. On day 25 of the experiment, six mice were randomly selected from both the pseudo-germ-free group and the conventional group to measure body weight, and fecal samples were collected for 16S rDNA sequencing to compare differences in gut microbiota, including operational taxonomic units (OTUs), Shannon index, Observed_species index, and phylogenetic diversity (PD whole tree). On day 39, body weight was measured in all groups, and the disease activity index (DAI) was evaluated. Colon length and colon wet weight were recorded. Histopathological changes in colonic tissues were assessed by hematoxylin and eosin (HE) staining, and histological injury scores were determined. In addition, the expression levels of mucopolysaccharides and mucin 2 (MUC2) in colonic tissues were measured. ResultsOn day 25, there was no significant difference in body weight between the pseudo-germ-free group and the conventional microbiota group (P>0.05). Compared to the conventional microbiota group, the pseudo-germ-free group showed significantly reduced OTUs, Shannon index, Observed_species index, and PD whole tree (P<0.01). On day 39, compared to the normal control group, mice in the conventional model group showed a significant decrease in the body weight and an increase in DAI, along with increased colon wet weight, shortened colon length, markedly elevated histopathological scores, and reduced expression of mucopolysaccharides and MUC2 (P<0.01). In contrast, no significant differences were observed in these parameters in the pseudo-germ-free model group (P>0.05). Compared to the conventional model group, the conventional QWD group showed significant improvement in all measured parameters, whereas the conventional mesalazine group showed reductions only in DAI scores and colon wet weight (P<0.01). No statistically significant differences were observed among the pseudo-germ-free model group, the pseudo-germ-free QWD group, and the pseudo-germ-free mesalazine group (P>0.05). ConclusionQWD may promote the reconstruction of the intestinal mucus barrier by regulating the intestinal flora, thereby exerting a therapeutic effect on acute UC.
2.Comparative Study on the Effects of Qingchang Wenzhong Decoction (清肠温中方) on the Intestinal Mucus Barrier in Conventional and Pseudo-Germ-Free Model Mice of Acute Ulcerative Colitis
Mingxu GAO ; Leilei LIU ; Lijie LU ; Jiali WANG ; Juncong HU ; Yangzhe LIN ; Qirui LIU ; Yue DONG ; Luyue WANG ; Zeyu XUE ; Junxiang LI ; Lei SHI
Journal of Traditional Chinese Medicine 2026;67(14):1528-1537
ObjectiveTo explore the mechanisms of Qingchang Wenzhong Decoction (清肠温中方, QWD) in treating ulcerative colitis (UC). MethodsA total of 100 male C57BL/6J mice were assigned to a pseudo-germ-free group (n=45) and a conventional group (n=55) using a random number table. Pseudo-germ-free model was established by giving antibiotic treatment and subsequently subjected to acute UC induction. After the model was successfully established, the 45 pseudo-germ-free acute UC mice were randomly allocated to a pseudo-germ-free model group, a pseudo-germ-free QWD group, and a pseudo-germ-free mesalazine group, with 15 mice in each group. The 55 conventional mice were randomly divided into a normal control group (n=10), as well as a conventional model group, a conventional QWD group, and a conventional mesalazine group, with 15 mice in each group. Except for the normal control group, acute UC models were established in all other groups. Mice in the control group received no intervention. From day 32 of the experiment, mice in the pseudo-germ-free QWD group and the conventional QWD group were administered with QWD granules at a dose of 9.71 g/(kg·d) by gavage with deionized water, while those in the pseudo-germ-free mesalazine group and the conventional mesalazine group were given mesalazine enteric-coated tablets at a dose of 151.67 mg/(kg·d) by gavage. Mice in the remaining groups received an equal volume of deionized water once daily for 7 consecutive days. On day 25 of the experiment, six mice were randomly selected from both the pseudo-germ-free group and the conventional group to measure body weight, and fecal samples were collected for 16S rDNA sequencing to compare differences in gut microbiota, including operational taxonomic units (OTUs), Shannon index, Observed_species index, and phylogenetic diversity (PD whole tree). On day 39, body weight was measured in all groups, and the disease activity index (DAI) was evaluated. Colon length and colon wet weight were recorded. Histopathological changes in colonic tissues were assessed by hematoxylin and eosin (HE) staining, and histological injury scores were determined. In addition, the expression levels of mucopolysaccharides and mucin 2 (MUC2) in colonic tissues were measured. ResultsOn day 25, there was no significant difference in body weight between the pseudo-germ-free group and the conventional microbiota group (P>0.05). Compared to the conventional microbiota group, the pseudo-germ-free group showed significantly reduced OTUs, Shannon index, Observed_species index, and PD whole tree (P<0.01). On day 39, compared to the normal control group, mice in the conventional model group showed a significant decrease in the body weight and an increase in DAI, along with increased colon wet weight, shortened colon length, markedly elevated histopathological scores, and reduced expression of mucopolysaccharides and MUC2 (P<0.01). In contrast, no significant differences were observed in these parameters in the pseudo-germ-free model group (P>0.05). Compared to the conventional model group, the conventional QWD group showed significant improvement in all measured parameters, whereas the conventional mesalazine group showed reductions only in DAI scores and colon wet weight (P<0.01). No statistically significant differences were observed among the pseudo-germ-free model group, the pseudo-germ-free QWD group, and the pseudo-germ-free mesalazine group (P>0.05). ConclusionQWD may promote the reconstruction of the intestinal mucus barrier by regulating the intestinal flora, thereby exerting a therapeutic effect on acute UC.
3.Thermal sensitization of acupoints in patients with knee osteoarthritis: A cross-sectional case-control study.
Jian-Feng TU ; Xue-Zhou WANG ; Shi-Yan YAN ; Yi-Ran WANG ; Jing-Wen YANG ; Guang-Xia SHI ; Wen-Zheng ZHANG ; Li-Na JIN ; Li-Sha YANG ; Dong-Hua LIU ; Li-Qiong WANG ; Bao-Hong MI
Journal of Integrative Medicine 2025;23(3):289-296
OBJECTIVE:
Varied acupoint selections represent a potential cause of the uncertainty surrounding the efficacy of acupuncture for knee osteoarthritis (OA). Skin temperature, a guiding factor for acupoint selection, may help to address this issue. This study explored thermal sensitization of acupoints used for the treatment of knee OA.
METHODS:
This cross-sectional case-control study enrolled cases aged 45-75 years with symptomatic knee OA and age- and gender-matched non-knee OA controls in a 1:1 ratio. All participants underwent infrared thermographic imaging. The primary outcome was the relative skin temperature of acupoint (STA), and the secondary outcome was the absolute STA of 11 acupoints. The Z test was used to compare the relative and absolute STAs between the groups. Principal component analysis was used to extract the common factors (CFs, acupoint cluster) in the STAs. A general linear model was used to identify factors affecting the STA in the knee OA cases. For the group comparisons of relative STA, P < 0.0045 (adjusted for 11 acupoints through Bonferroni correction) was considered to indicate statistical significance. For other analyses, P < 0.05 was used as the threshold for statistical significance.
RESULTS:
The analysis included 308 participants, consisting of 151 cases (mean age: [64.58 ± 6.67] years; male: 25.83%; mean body mass index: [25.70 ± 3.16] kg/m2) and 157 controls (mean age: [63.37 ± 5.96] years; male: 26.11%; mean body mass index: [24.47 ± 2.84] kg/m2). The relative STAs of ST34 (P = 0.0001), EX-LE2 (P < 0.0001), EX-LE5 (P = 0.0006), SP10 (P < 0.0001), BL40 (P = 0.0012) and GB39 (P = 0.0037) were higher in the knee OA group. No difference was found in the STAs of ST35, ST36, SP9, GB33 and GB34. Four CFs were identified for relative STA in both groups. The acupoints within each CF were consistent between the groups. The mean values of the relative STAs across each CF were higher in the knee OA group. In the knee OA cases, no factors were observed to affect the relative STA, while age and gender were found to affect the absolute STA.
CONCLUSION
Among patients with knee OA, thermal sensitization occurs in the acupoints of the lower extremity, exhibiting localized and regional thermal consistencies. The thermally sensitized acupoints that we identified in this study, ST34, SP10, EX-LE2, EX-LE5, GB39 and BL40, may be good choices for the acupuncture treatment of knee OA. Please cite this article as: Tu JF, Wang XZ, Yan SY, Wang YR, Yang JW, Shi GX, Zhang WZ, Jing LN, Yang LS, Liu DH, Wang LQ, Mi BH. Thermal sensitization of acupoints in patients with knee osteoarthritis: A cross-sectional case-control study. J Integr Med. 2025; 23(3): 289-296.
Humans
;
Osteoarthritis, Knee/physiopathology*
;
Male
;
Cross-Sectional Studies
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Middle Aged
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Female
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Acupuncture Points
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Case-Control Studies
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Aged
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Skin Temperature
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Acupuncture Therapy
4.Serum Lipidomics Profiling to Identify Potential Biomarkers of Ischemic Stroke: A Pilot Study in Chinese Adults.
Ji Jun SHI ; Zu Jiao NIE ; Shu Yao WANG ; Hao ZHANG ; Xin Wei LI ; Jia Ling YAO ; Yi Bing JIN ; Xiang Dong YANG ; Xue Yang ZHANG ; Ming Zhi ZHANG ; Hao PENG
Biomedical and Environmental Sciences 2025;38(8):918-925
OBJECTIVE:
Lipid oxidation is involved in the pathogenesis of atherosclerosis and may be contribute to the development of Ischemic stroke (IS). However, the lipid profiles associated with IS have been poorly studied. We conducted a pilot study to identify potential IS-related lipid molecules and pathways using lipidomic profiling.
METHODS:
Serum lipidomic profiling was performed using LC-MS in 20 patients with IS and 20 age- and sex-matched healthy controls. Univariate and multivariate analyses were simultaneously performed to identify the differential lipids. Multiple testing was controlled for using a false discovery rate (FDR) approach. Enrichment analysis was performed using MetaboAnalyst software.
RESULTS:
Based on the 294 lipids assayed, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) models were used to distinguish patients with IS from healthy controls. Fifty-six differential lipids were identified with an FDR-adjusted P less than 0.05 and variable influences in projection (VIP) greater than 1.0. These lipids were significantly enriched in glycerophospholipid metabolism (FDR-adjusted P = 0.009, impact score = 0.216).
CONCLUSIONS
Serum lipid profiles differed significantly between patients with IS and healthy controls. Thus, glycerophospholipid metabolism may be involved in the development of IS. These results provide initial evidence that lipid molecules and their related metabolites may serve as new biomarkers and potential therapeutic targets for IS.
Humans
;
Pilot Projects
;
Lipidomics
;
Male
;
Female
;
Biomarkers/blood*
;
Middle Aged
;
Ischemic Stroke/blood*
;
Aged
;
China
;
Lipids/blood*
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Adult
;
Case-Control Studies
;
East Asian People
5.Stent location and effectiveness in different types of iliac vein compression
Song XUE ; Dong YAN ; Huihua SHI ; Zhen ZHAO ; Xinwu LU ; Minyi YIN
Journal of Surgery Concepts & Practice 2025;30(2):159-164
Objective To investigate the stent location and effectiveness in different types of iliac vein compression.Methods A retrospective analysis was conducted on patients with left iliac vein compression who underwent stent implantation at our department from June 2021 to December 2023. Based on the location of compression, patients were categorized into high, classical, and low types. The patients’ general information, lesion characteristics, stent details, stent patency, and clinical outcomes were analyzed and followed up.Results A total of 242 patients were included in this study. And 90. 9% (220 cases) were the classical type, 5.4% (13 cases) were the high type, and 3.7% (9 cases) were the low type. There was no significant difference in age distribution, gender ratio and comorbidities among the three groups. The length of inferior vena cava protruding from the high type was significantly longer than that of the classical type[(3.1±0.8) cm vs. (1.6±1.0) cm, P<0.001), but significantly shorter in the low type[(-0.7±2.4) cm vs. (1.6±1.0) cm, P<0.001). The mean follow-up time was (18.0±7.0) months. The stent patency rate was 97.0% at 1 year and 92.8% at 2 years postoperation. The clinical symptoms of the three groups were significantly relieved. No thrombosis was found in right limbs.Conclusions There is a significant difference in the location of the stent tip between classical and non-classical compression types. Complete coverage of lesion by the stent should be the prerequisite for stent placement in treatment of all three types. Small sample, short-term follow-up data suggested that stents can also relieve clinical symptoms and have good patency rate in the high compression type and the low compression type.
6.Effects and mechanisms of soluble guanylate cyclase stimulator sGC003F on chronic heart failure induced by aortic arch constriction in mice
Xin CHENG ; Shuo LI ; Jinshui ZHANG ; Yingxian SHI ; Yang ZHANG ; Rui XUE ; Jincao LI ; Huajin DONG ; Zhibing ZHENG ; Youzhi ZHANG ; Henglin WANG
Chinese Journal of Pharmacology and Toxicology 2025;39(1):25-35
OBJECTIVE To investigate the effect and mechanism of soluble guanylate cyclase stimulator sGC003F on cardiac function in mice with chronic heart failure(CHF).METHODS C57BL/6J male mice were randomly divided into the sham operation(sham)group,transverse aortic constriction induced CHF mouse model group,model+veliciguat(Ver,3 mg·kg-1)group(positive control)and model+sGC003F(3 and 10 mg·kg-1)group.Four weeks after modeling,drugs were ig given,once a day,for 28 d.Echocardiography was used to measure the changes in cardiac function,and the myocardial hypertrophy related indexes were calculated.The levels of serum N-terminal pro-brain natriuretic peptide(NT-pro-BNP),N-terminal pro-atrial natriuretic peptide(NT-pro-ANP),soluble guanylate cyclase(sGC),cyclic guanosine monophosphate(cGMP)and inflammatory factors interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and IL-1β were detected by ELISA.The pathological changes of left heart tissue were observed with HE and Masson staining.Image was used to analyze the percentage of fibrosis in cardiac tissus stained with Masson.The activity of superoxide dismutase(SOD),content of malondialdehyde(MDA)in myocardial tissue,and level of nitric oxide(NO)in serum were detected by biochemical detection kits.The protein expression levels of p-mammalian target of rapamycin(p-mTOR),p-protein kinase B(p-Akt),TNF-α and IL-6 in cardiac tissue were detected by Western blotting.RESULTS Com-pared with the sham group,the left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFs)in the model group decreased significantly(P<0.01),the cardiac structure changed significantly,the percentage of myocardial fibrosis increased significantly(P<0.05),so were serum NT-pro-BNP and NT-pro-ANP levels(P<0.01).Compared with the model group,the above indexes of the model+Ver group and the model+sGC003F 3 mg·kg-1 group were significantly improved(P<0.05,P<0.01).The sGC003F 10 mg·kg-1 group had a significant improvement in LVEF,LVFs,and NT-pro-BNP(P<0.01).Compared with the sham group,the serum levels of NO,sGC and cGMP in the model group decreased significantly(P<0.05,P<0.01).Compared with the model group,the serum levels of NO,sGC and cGMP were significantly increased in the model+sGC003F 3 mg·kg-1 group(P<0.01),but only serum cGMP levels were significantly increased in model+Ver and model+sGC003F 10 mg·kg-1 groups(P<0.01).Compared with the sham group,the serum levels of TNF-α,IL-1β and IL-6 in the model group were significantly increased(P<0.05,P<0.01).Compared with the model group,the serum levels of TNF-α,IL-1β and IL-6 were significantly decreased in the model+sGC003F 3 mg·kg-1 group(P<0.05,P<0.01),and only the TNF-α level was significantly decreased in the model+sGC003F 10 mg·kg-1 group(P<0.01).Compared with the sham group,the SOD activity of the model group was significantly decreased(P<0.01),but the MDA content significantly increased(P<0.01).Compared with the model group,SOD and MDA were significantly improved in the model+sGC003F 3 mg·kg-1 group(P<0.05,P<0.01),but in the model+Ver group only the SOD activity significantly increased(P<0.05).Western blotting showed that the expressions of p-mTOR,p-Akt,TNF-α and IL-6 protein in myocardial tissue of the model group were significantly higher than in the sham group(P<0.05).Compared with the model group,the expressions of the above proteins in the model+sGC003F 3 mg·kg-1 group were significantly decreased(P<0.05,P<0.01),so were the expressions of TNF-α protein in the model+sGC003F 10 mg·kg-1 group and model+Ver group(P<0.01).CONCLUSION sGC003F can improve cardiac function,and reduce myocardial fibrosis in CHF model mice,which may be related to the inhibition of myocardial oxidative stress and inflammation,and the regulation of NO/sGC/cGMP and AKT/mTOR signaling pathways.
7.The postoperative care of a patient with uremic complicated by ventricular tachycardia undergoing catheter radiofrequency ablation therapy
Yanli DONG ; Qingyan SHI ; Junlin XUE ; Qianqian ZUO ; Hui JU ; Junying QIAN
Chinese Journal of Nursing 2025;60(18):2275-2278
To summarize the nursing experience of a patient with uremia on maintenance hemodialysis complicated by recurrent ventricular tachycardia and treated with transcatheter radiofrequency ablation.Key nursing interventions included:dynamically assessing the patient's coagulation and bleeding status,being vigilant against the occurrence of deep vein thrombosis,and preventing local and major organ bleeding;implementing goal-oriented volume management strategies to prevent electrolyte disorders;the strengthened management of vascular access to reduce risks of stenosis or occlusion in the arteriovenous fistula;conducting precise assessment and comprehensive intervention to reduce the patient's psychological and mental burden.After careful treatment and nursing care,the patient was stable and discharged on the 6th postoperative day.During the 2-month outpatient follow-up,cardiac function indicators were normal,and the fistula was unobstructed,and the patient recovered well.
8.Effects of different acupuncture and moxibustion methods on synovial hypoxia and fibrosis in knee osteoarthritis model rats
Lingling QIN ; Xiaoqin ZHANG ; Qi DONG ; Mengting SHI ; Pingju XUE ; Haijiao XING ; Jing XU
Journal of Acupuncture and Tuina Science 2025;23(5):394-402
Objective:To investigate the effects of moxibustion,filiform needle acupuncture,and electroacupuncture on synovial hypoxia and expression of synovial fibrosis markers[transforming growth factor(TGF)-β and collagen alpha-1(Ⅰ)chain(COL1A1)]in knee osteoarthritis(KOA)rats.Methods:Fifty Sprague-Dawley rats were randomly divided into a normal group,a model group,a moxibustion group,an acupuncture group,and an electroacupuncture group,with 10 rats in each group.Except for the normal group,rats in the other four groups were injected with sodium iodoacetate into the right knee joint cavity to establish KOA models.The right Futu(ST32)and Zusanli(ST36)were selected in the moxibustion,acupuncture,and electroacupuncture groups to perform the mild moxibustion with moxa sticks,filiform needle acupuncture,or electroacupuncture,respectively.Each intervention lasted 15 min,once every other day,and continued for 4 weeks,with a total of 14 interventions.The Lequesne score was used to evaluate the degree of knee dysfunction before and after intervention,and the rat's right knee joint diameter was measured to evaluate the degree of knee swelling.Morphological changes of the right knee joint synovial tissue were observed using hematoxylin-eosin staining.Hypoxia probe immunofluorescence staining was used to observe the degree of synovial hypoxia in rats.Western blotting was used to detect the protein expression levels of hypoxia-inducible factor(HIF)-1α,TGF-β,and COL1A1 in the synovial tissue of the right knee joints.Results:Before intervention,the Lequesne score and the right knee joint diameter in the other four groups increased significantly compared to the normal group(P<0.01).After intervention,the Lequesne score and the right knee joint diameter increased significantly in the model group compared to the normal group(P<0.01)together with worsened cartilage deformation and osteophytes,inflammatory cell infiltration and fibrosis in synovial tissue,and hypoxia degree,and the expression levels of HIF-1α,TGF-β,and COL1A1 proteins in the knee joint synovial tissue of rats increased(P<0.01).The Lequesne score and the right knee joint diameter decreased significantly(P<0.01 or P<0.05),the cartilage morphology was normal without obvious osteophytes,and the hypoxia degree reduced in the three intervention groups compared to the model group.Among them,the moxibustion group had the most notable improvement in hypoxia,synovial tissue fibrosis,angiogenesis,and inflammatory cell infiltration.The expression levels of HIF-1α,TGF-β,and COL1A1 proteins decreased in the moxibustion and electroacupuncture groups(P<0.01),and the expression levels of HIF-1α and TGF-β proteins were decreased in the acupuncture group(P<0.01).Conclusion:Moxibustion,filiform needle acupuncture,and electroacupuncture improve knee joint function and reduce knee joint swelling in KOA rats.The mechanism may be to improve synovial fibrosis of the knee joint by regulating HIF-1α;moxibustion has the best effect on improving hypoxia among the three interventions,but the effect of filiform needle acupuncture on COL1A1 is not significant.
9.Comparison of chemical constituents in traditional decoction and formula granule decoction of Wendan Decoction
Tan XUE ; Man-wen XU ; Xue-hua FAN ; Feng-yu DONG ; Yan MIAO ; Jia-ning SUN ; Jun-han SHI ; Lu ZHANG ; Jing YAO ; Rui-xin LIU
Chinese Traditional Patent Medicine 2025;47(2):384-394
AIM To compare the chemical constituents in traditional decoction and formula granule decoction of classical famous prescription Wendan Decoction.METHODS The HPLC fingerprints were established,after which the contents of adenosine,synephrine,liquiritin,naringin,hesperidin,6-gingerol and adenosine cyclophosphate were determined,cluster analysis,principal component analysis and multidimensional scaling analysis were adopted in the investigation of component differences,and the equivalent of formula granules was adjusted.RESULTS The similarities of HPLC fingerprints for 10 batches of traditional decoctions were higher than those of HPLC fingerprints for 9 batches of formula granule decoctions(P<0.01).Adenosine,synephrine,liquiritin,hesperidin and cyclic adenosine monophosphate demonstrated higher contents in traditional decoctions than those in formula granule decoctions(P<0.05),6-gingerol displayed lower content than that in the latter produced by manufacturers A,C(P<0.05),which was higher than that in the latter produced by manufacturer B(P<0.01).Various batches of traditional decoctions and formula granule decoctions could be obviously distinguished,adenosine,synephrine and hesperidin exhibited great influences on the classification of principal component analysis,and the quality of formula granule decoctions produced by manufacturer C was closer to that of traditional decoctions.After equivalent correction,the contents of various constituents in formula granule decoctions produced by manufacturers A,C showed no significant differences as compared with those in traditional decoction(P>0.05).CONCLUSION The formula granules of Wendan Decoction from different manufacturers exist quality differences,so the preparation process and extraction process of this preparation should be optimized to improve quality,and equivalent ratio should be adjusted according to actual requirements to ensure its scientific and rational clinical application.
10.Mechanism of baicalin combined with heat stimulation in treating acute lymphoblastic leukemia based on network pharmacology and in vitro experimental verification
Zi-ru LIU ; Zhu-yun SUN ; Ping-liang GE ; Ran SHI ; Xiao-yun LIU ; Dong-xue YE ; Guo-ying ZHANG ; Rong RONG ; Yong YANG
Chinese Pharmacological Bulletin 2025;41(6):1167-1176
Aim To explore the mechanism of baicalin combined with heat stimulation in treating acute lym-phoblastic leukemia(ALL)based on network pharma-cology and in vitro experiments.Methods The CCK-8 assay was used to screen the suitable conditions for heat stimulation to interfere ALL cell lines Jurkat,CCRF-CEM,Hut-78 and a normal lymphocyte HMy2.CIR,and the effects of baicalin combined with heat stimulation on the proliferation of three ALL cell lines and a normal lymphocyte were tested.The key targets of baicalin combined with fever stimulation for the treatment of ALL were obtained based on network phar-macological analysis,and the potential mechanisms were predicted by gene ontology(GO)annotation and kyoto encyclopedia of genes and genomes(KEGG)en-richment.The expression levels of TNF-α,AKT1,TYMS and CASP3 mRNA in ALL cell lines Jurkat and CCRF-CEM were examined by RT-qPCR with baicalin alone and baicalin combined with heat stimulation.Results The optimal conditions for heat stimulation to intervene ALL cells were 41 ℃ for 24 h,and heat stimulation combined with baicalin synergistically inhibited the growth of ALL cell lines and effectively reduced the cy-totoxicity of baicalin.Based on the network pharmaco-logical analysis,55 intersecting targets of baicalin with ALL diseases and 77 intersecting targets of baicalin with fever were obtained.The results of GO annotation and KEGG enrichment suggested that baicalin com-bined with fever stimulation to intervene ALL might be associated with influencing intracellular reactive oxygen species metabolism,DNA transcription and apoptotic processes involved in cysteine enzymes.Apoptosis,TNF and IL-17 signaling pathways were the key pathways for baicalin combined with heat stimulation in treating ALL.Under heat stimulation at 41 ℃ using SDHA gene as housekeeping gene,in vitro experiments showed that baicalin significantly up-regulated the expression of TNF-α and CASP3,and down-regulated the expression of TYMS in ALL cells.Conclusions Based on net-work pharmacologic analyses and in vitro experiments,baicalin combined with heat stimulation can regulate TNF-α and CASP3 gene levels in ALL cells and de-stroy cellular structure to promote cell apoptosis,thus synergistically treating ALL.

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