1.Prevalence of human T-lymphocyte leukemia virus among blood donors in Guangzhou
Yuxiao LI ; Xia RONG ; Bo HE ; Rongsong DU ; Zhengang SHAN ; Qiao LIAO
Chinese Journal of Blood Transfusion 2025;38(2):222-226
[Objective] To analyze the prevalence of human T-lymphocyte leukemia virus (HTLV) among blood donors in Guangzhou from 2016 to 2021, and provide a basis for blood collection and supply management in this region. [Methods] A total of 2 116 951 voluntary blood donors were screened for anti-HTLV by enzyme-linked immunosorbent assay (ELISA) from March 2016 to December 2021 in Guangzhou, and the reactive cases were further confirmed by Western blotting (WB). Qualitative data were analyzed by χ2 with spss19 software. The trend of the total positive rate of HTLV confirmation test by WB from 2016 to 2021 was analyzed with the Joinpoint software, and the annual percent change (APC) was used to determine whether the trend changes were statistically significant. [Results] From March 2016 to December 2021, the total positive rate for anti-HTLV by ELISA among voluntary blood donors in Guangzhou was 0.019 7% (416/ 2116 951), and the WB confirmed positive rate was 0.001 1% (23/2 116 951). The total positive rate of HTLV among individual voluntary blood donors in the six main districts (0.002 12%, 19/895 301) was higher than that among group voluntary blood donors (0.000 32%, 3/951 947) (P<0.05). There was no significant difference in the total positive rate of HTLV confirmation between the six main districts (0.001 19%) and the three non-main districts (0.000 37%) (P>0.05). The trend of the total positive rate of HTLV infection in the six main districts and the Guangzhou area(including the six main districts and three non-main districts) showed no significant increase or decrease. [Conclusion] The prevalence of HTLV among blood donors in Guangzhou remains at a low level.
2.Effect of the nitroglycerin-controlled low central venous pressure technique on cerebral metabolic markers and cerebral blood oxygen saturation in patients undergoing laparoscopic hepatectomy for liver cancer
Bo WANG ; Xia FU ; Conghai LYU ; Chunfang YIN ; Qiyuan WU
Journal of Clinical Hepatology 2025;41(3):478-484
ObjectiveTo investigate the effect of the nitroglycerin-controlled low central venous pressure (CLCVP) technique on brain metabolic markers and cerebral blood oxygen saturation in patients undergoing laparoscopic hepatectomy for liver cancer, and to reduce the risk of neurological complications. MethodsA total of 105 patients who underwent elective laparoscopic hepatectomy for liver cancer in Haikou Hospital Affiliated to Xiangya Hospital of Central South University from April 2020 to May 2023 were enrolled and randomly divided into CLCVP group with 54 patients and non-CLCVP group with 51 patients. The patients in the CLCVP group were treated with the nitroglycerin CLCVP technique during surgery, while those in the non-CLCVP group were given conventional surgical treatment. The two groups were compared in terms of the following indicators: perioperative indicators; hemodynamic parameters and cerebral oxygen metabolism before anesthesia induction (T0), at 5 minutes after anesthesia induction (T1), at 5 minutes after the beginning of liver parenchyma dissection (T2), at 5 minutes after the end of hepatectomy (T3), and immediately after the end of surgery (T4); the changes in liver function parameters after surgery; the incidence rate of adverse reactions. The independent-samples t test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups; the analysis of variance with repeated measures was used for comparison between multiple time points. ResultsCompared with the non-CLCVP group, the CLCVP group had significantly lower intraoperative blood loss and intraoperative fluid infusion volume (t=5.408 and 7.220, both P<0.05), while there were no significant differences between the two groups in time of operation, anesthesia time, extubation time, resuscitation time and intraoperative urine volume (all P>0.05). Compared with the data at T0, both groups had significant reductions in mean arterial pressure, heart rate, and central venous pressure during surgery (all P<0.05), and compared with the non-CLCVP group, the CLCVP group had significantly lower mean arterial pressure and central venous pressure (P<0.05) and a significantly higher heart rate (P<0.05) at T2 and T3. Compared with the data at T0, both groups had a significant reduction in Ca-jvDO2 at T2 — T4 time points (all P<0.05), while there was no significant difference in Ca-jvDO2 between the two groups at each time point (all P>0.05). Compared with the data at T0, the CLCVP group had a significant reduction in rSO2 at T2 — T4 time points (all P<0.05), and the CLCVP group had a significantly lower level of rSO2 than the non-CLCVP group at T2 — T3 time points (both P<0.05); there were no significant changes in CERO2 and Djv-aBL in either group at each time point (all P>0.05). At 3 and 7 days after surgery, both groups had significant increases in the liver function parameters of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TBil) (all P<0.05), and the CLCVP group had significantly lower levels of AST and ALT than the non-CLCVP group (all P<0.05); there was no significant difference in TBil between the two groups at each time point (all P>0.05). There was no significant difference in the incidence rate of perioperative complications between the two groups (χ2=0.729, P=0.394). ConclusionThe application of the nitroglycerin CLCVP technique in laparoscopic hepatectomy for liver cancer can reduce the amount of intraoperative blood loss in patients, but it is necessary to further enhance the monitoring of cerebral blood oxygen saturation during surgery, so as to reduce the risk of neurological complications as much as possible.
3.Component Analysis of Anmeidan and Its Mechanism in Regulating ERK1/2/MNK/ELF4E Signaling Pathway to Improve Circadian Rhythm in Insomnia Rats
Yi GAO ; Bo XU ; Jing XIA ; Linlin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):44-53
ObjectiveTo identify the main chemical constituents of Anmeidan (AMD) and to explore the mechanism of AMD in regulating the extracellular signal-regulated kinase 1/2 (ERK1/2)/mitogen-activated protein kinase (MAPK)-interacting serine/threonine-protein kinase (MNK)/eukaryotic translation initiation factor 4E (eIF4E) signaling pathway to improve circadian rhythm disturbances in insomnia rats. MethodsThe main chemical constituents of AMD were identified using ultra-high-performance liquid chromatography-linear ion trap-electrostatic orbital trap mass spectrometry (UPLC-LTQ/Orbitrap/MS) in combination with reference standards. Sixty male Sprague-Dawley (SD) rats were randomly divided into control, model, melatonin, and AMD low-, medium-, and high-dose groups, with 10 rats in each group. Except for the control group, all rats were administered p-chlorophenylalanine via intraperitoneal injection to establish an insomnia model. The activity-rest rhythm of rats was assessed using the open field test and circadian rhythm test. Hematoxylin-eosin (HE) staining and Nissl staining were used to observe structural changes in hypothalamic neurons. Immunofluorescence, real-time quantitative polymerase chain reaction (Real-time PCR), and Western blot analysis were employed to detect mRNA and protein expression levels of ERK1/2, MNK, and eIF4E in the hypothalamus. ResultsA total of 50 chemical components, including flavonoids, phenylpropanoids, triterpenoid saponins, alkaloids, and lignans, were identified in AMD. Compared with the control group, the model group exhibited significantly increased total distance traveled, average speed, central area residence time, and cumulative rearing time (P<0.01), as well as prolonged cumulative activity time and total activity time in both light and dark phases (P<0.01). Hypothalamic neurons in the model group were sparsely arranged, reduced in number, and exhibited nuclear disappearance or nucleolar rupture, with a significantly increased apoptosis index (P<0.01). The cytoplasm appeared turbid, Nissl body staining was lighter, and the Nissl body apoptosis index was significantly increased (P<0.01). The mRNA expression levels of ERK1/2, MNK, and eIF4E were significantly decreased (P<0.01), along with a significant reduction in protein expression levels of ERK1/2, phosphorylated ERK1/2 (p-ERK1/2), MNK, phosphorylated MNK (p-MNK), eIF4E, and phosphorylated eIF4E (p-eIF4E) (P<0.01). Compared with the model group, the total distance, average speed, central area residence time and body upright cumulative time of the AMD high-dose group were significantly reduced (P<0.01). The total distance, average speed and body upright cumulative time of the AMD medium-dose group were significantly reduced (P<0.01). The cumulative time of light activity and total time of activity in each dose group of AMD were significantly shortened (P<0.01). The cumulative time of dark activity in the high-dose group of AMD was prolonged (P<0.01). The neurons in the middle and high dose groups of AMD were closely arranged, the number of neurons increased, and the apoptosis index of hypothalamic cells decreased significantly (P<0.05, P<0.01). The cytoplasm of the low, middle and high dose groups of AMD was clear, the color of Nissl body became darker, and the apoptosis index of Nissl body decreased significantly (P<0.01). The expression of ERK1/2, MNK and eIF4E mRNA and protein in the hypothalamus of the middle and high dose groups of AMD increased significantly (P<0.05, P<0.01). ConclusionAMD primarily contains 50 chemical constituents, including flavonoids, phenylpropanoids, and triterpenoid saponins. It exhibits a "synergistic enhancement" effect through multiple components and multiple pathways to improve insomnia. AMD ameliorates circadian rhythm disturbances in p-chlorophenylalanine-induced insomnia rats by upregulating ERK1/2/MNK/eIF4E signaling pathway-related proteins.
4.Protective Effect and Mechanism of Anmeidan against Neuronal Damage in Rat Model of Sleep Deprivation Based on Hippocampal Neuroinflammation
Guangjing XIE ; Zixuan XU ; Junlu ZHANG ; Jian ZHANG ; Jing XIA ; Bo XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):65-71
ObjectiveTo investigate the effects of Anmeidan (AMD) on neuroinflammation in the hippocampus of sleep-deprived rats. MethodsSD rats were randomly divided into four groups (n = 10 per group): control group, model group, AMD group, and melatonin group. A sleep deprivation model was established using the modified multiple platform water environment method. The AMD group received AMD at a dose of 18.18 g·kg-1·d-1, the melatonin group received melatonin at 100 mg·kg-1·d-1, and the control and model groups were given an equal volume of pure water. All treatments were administered by gavage for four weeks. Spontaneous activity was assessed using an animal behavior video system. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). Hippocampal pyramidal neuron morphology was examined using hematoxylin-eosin (HE) staining, and ultrastructural changes of hippocampal neurons were observed via transmission electron microscopy. Immunofluorescence was used to detect the expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in the hippocampus. Western blot analysis was performed to measure the expression of nuclear factor-κB (NF-κB), phosphorylated NF-κB (p-NF-κB), NOD-like receptor protein 3 (NLRP3), and Caspase-1 proteins. ResultsCompared with the control group, the model group showed a significant increase in activity duration and frequency (P<0.01), increased hippocampal pyramidal cell structural damage and decreased cell count, aggravated hippocampal ultrastructural damage, mitochondrial cristae disruption, and exacerbated vacuolization. The expression of p-NF-κB p65, NLRP3, and Caspase-1 proteins was upregulated, serum IL-1β, IL-6, and TNF-α levels were significantly elevated (P<0.01), and the fluorescence intensity of BDNF and NGF proteins was significantly reduced (P<0.01). Compared with the model group, the AMD group showed a significant reduction in activity duration and frequency (P<0.01), increased hippocampal pyramidal cell count with reduced structural damage, alleviated hippocampal ultrastructural damage, significantly downregulated p-NF-κB p65, NLRP3, and Caspase-1 protein expression (P<0.01), decreased serum IL-1β, IL-6, and TNF-α levels (P<0.01), and significantly increased the fluorescence intensity of BDNF and NGF proteins (P<0.01). ConclusionAnmeidan alleviates hippocampal neuronal damage in sleep-deprived rats, potentially by downregulating the NLRP3 signaling pathway, reducing inflammatory cytokine release, and increasing neurotrophic factor levels.
5.Alleviation of Ulcerative Colitis by Shaoyaotang via Inhibiting Glycolysis Through SIRT6/HIF-1α Pathway
Yiling XIA ; Hui CAO ; Dongsheng WU ; Bo ZOU ; Erle LIU ; Yiwen WANG ; Shaijin JIANG ; Yiqian YU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):10-19
ObjectiveTo investigate the role of silent information regulatory protein (SIRT6)/hypoxia-inducible factor-1α (HIF-1α) pathway in regulating the reprogramming of glucose metabolism in ulcerative colitis (UC) and the mechanism of intervention of Shaoyaotang. MethodsForty-eight c57bL/6 mice were randomly divided into a blank group, a model group, a Mesalazine group (0.42 g·kg-1), a Shaoyaotang group (31.08 g·kg-1), an inhibitor group (OSS-128167, 50 mg·kg-1), and an inhibitor + Shaoyaotang group (50 mg·kg-1 OSS-128167 + 31.08 g·kg-1 Shaoyaotang). A UC model was established by the administration of 2.5% dextran sulfate sodium (DSS) solution for mice in other groups for 7 d, except for the blank group. The mice in each group were treated with saline, Mesalazine, Shaoyaotang, inhibitor, and inhibitor + Shaoyaotang, respectively, for 7 d. The mice were necropsied 24 h after the last administration of the drug. The blood was collected from the orbital region, and colon tissue was taken. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in colon tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to detect serum interleukin (IL)-10, IL-17, and IL-6 levels. A biochemical method was used to detect glucose and lactate dehydrogenase A (LDHA) levels. Immunohistochemistry (IHC) was employed to detect IL-22 and transforming growth factor-β1 (TGF-β1) levels in colon tissue, and Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect relative protein and mRNA expressions of SIRT6, HIF-1α, and LDHA. ResultsCompared with those of the blank group, disease activity index (DAI) scores of mice in the model group and inhibitor group were significantly increased (P<0.01). The length of colon tissue was significantly shortened, and colon tissue was congested and eroded. The pathohistological scores were significantly increased (P<0.01). The levels of serum inflammatory factors IL-17 and IL-6 were significantly elevated, and the levels of IL-10 were significantly decreased (P<0.01). The protein expressions of IL-22 and TGF-β1 were significantly reduced in colon tissue (P<0.01). The relative protein and mRNA expressions of SIRT6 were significantly decreased (P<0.01), and the relative protein and mRNA expressions of HIF-1α and LDHA and the contents of glucose and lactate were significantly elevated (P<0.01). The level of inflammation in the colon of the mice in the inhibitor group was more severe than that in the model group (P<0.01). Compared with the model group, the Mesalazine group, the Shaoyaotang group, and the inhibitor + Shaoyaotang group showed reduced colonic injury, significant decrease in serum IL-17 and IL-6, significant increase in IL-10 (P<0.01), significant increase in the protein expressions of IL-22 and TGF-β1 in colon tissue (P<0.01), significant increase in the protein expressions of SIRT6 and the relative mRNA expressions (P<0.01), and significant reduction in the protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate (P<0.01). Compared with those in the Shaoyaotang group, the serum IL-17 and IL-6 were significantly increased, and IL-10 was significantly decreased in the inhibitor + Shaoyaotang group (P<0.01). The protein expressions of IL-22 and TGF-β1 in colon tissue were significantly decreased (P<0.01). The expressions of SIRT6 protein and the relative mRNA expressions were significantly decreased (P<0.01). The protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate were significantly elevated (P<0.01). However, the difference between the Shaoyaotang group and the Mesalazine group was not significant. ConclusionShaoyaotang can effectively treat DSS-induced mice with UC through the SIRT6/HIF-1α pathway, and its mechanism of action may be related to the regulation of the SIRT6/HIF-1α pathway and glucose metabolism reprogramming and the inhibition of glycolysis.
6.Alleviation of Ulcerative Colitis by Shaoyaotang via Inhibiting Glycolysis Through SIRT6/HIF-1α Pathway
Yiling XIA ; Hui CAO ; Dongsheng WU ; Bo ZOU ; Erle LIU ; Yiwen WANG ; Shaijin JIANG ; Yiqian YU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):10-19
ObjectiveTo investigate the role of silent information regulatory protein (SIRT6)/hypoxia-inducible factor-1α (HIF-1α) pathway in regulating the reprogramming of glucose metabolism in ulcerative colitis (UC) and the mechanism of intervention of Shaoyaotang. MethodsForty-eight c57bL/6 mice were randomly divided into a blank group, a model group, a Mesalazine group (0.42 g·kg-1), a Shaoyaotang group (31.08 g·kg-1), an inhibitor group (OSS-128167, 50 mg·kg-1), and an inhibitor + Shaoyaotang group (50 mg·kg-1 OSS-128167 + 31.08 g·kg-1 Shaoyaotang). A UC model was established by the administration of 2.5% dextran sulfate sodium (DSS) solution for mice in other groups for 7 d, except for the blank group. The mice in each group were treated with saline, Mesalazine, Shaoyaotang, inhibitor, and inhibitor + Shaoyaotang, respectively, for 7 d. The mice were necropsied 24 h after the last administration of the drug. The blood was collected from the orbital region, and colon tissue was taken. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in colon tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to detect serum interleukin (IL)-10, IL-17, and IL-6 levels. A biochemical method was used to detect glucose and lactate dehydrogenase A (LDHA) levels. Immunohistochemistry (IHC) was employed to detect IL-22 and transforming growth factor-β1 (TGF-β1) levels in colon tissue, and Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect relative protein and mRNA expressions of SIRT6, HIF-1α, and LDHA. ResultsCompared with those of the blank group, disease activity index (DAI) scores of mice in the model group and inhibitor group were significantly increased (P<0.01). The length of colon tissue was significantly shortened, and colon tissue was congested and eroded. The pathohistological scores were significantly increased (P<0.01). The levels of serum inflammatory factors IL-17 and IL-6 were significantly elevated, and the levels of IL-10 were significantly decreased (P<0.01). The protein expressions of IL-22 and TGF-β1 were significantly reduced in colon tissue (P<0.01). The relative protein and mRNA expressions of SIRT6 were significantly decreased (P<0.01), and the relative protein and mRNA expressions of HIF-1α and LDHA and the contents of glucose and lactate were significantly elevated (P<0.01). The level of inflammation in the colon of the mice in the inhibitor group was more severe than that in the model group (P<0.01). Compared with the model group, the Mesalazine group, the Shaoyaotang group, and the inhibitor + Shaoyaotang group showed reduced colonic injury, significant decrease in serum IL-17 and IL-6, significant increase in IL-10 (P<0.01), significant increase in the protein expressions of IL-22 and TGF-β1 in colon tissue (P<0.01), significant increase in the protein expressions of SIRT6 and the relative mRNA expressions (P<0.01), and significant reduction in the protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate (P<0.01). Compared with those in the Shaoyaotang group, the serum IL-17 and IL-6 were significantly increased, and IL-10 was significantly decreased in the inhibitor + Shaoyaotang group (P<0.01). The protein expressions of IL-22 and TGF-β1 in colon tissue were significantly decreased (P<0.01). The expressions of SIRT6 protein and the relative mRNA expressions were significantly decreased (P<0.01). The protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate were significantly elevated (P<0.01). However, the difference between the Shaoyaotang group and the Mesalazine group was not significant. ConclusionShaoyaotang can effectively treat DSS-induced mice with UC through the SIRT6/HIF-1α pathway, and its mechanism of action may be related to the regulation of the SIRT6/HIF-1α pathway and glucose metabolism reprogramming and the inhibition of glycolysis.
7.A Case Report of Pachydermoperiostosis by Multidisciplinary Diagnosis and Treatment
Jie ZHANG ; Yan ZHANG ; Li HUO ; Ke LYU ; Tao WANG ; Ze'nan XIA ; Xiao LONG ; Kexin XU ; Nan WU ; Bo YANG ; Weibo XIA ; Rongrong HU ; Limeng CHEN ; Ji LI ; Xia HONG ; Yan ZHANG ; Yagang ZUO
JOURNAL OF RARE DISEASES 2025;4(1):75-82
A 20-year-old male patient presented to the Department of Dermatology of Peking Union Medical College Hospital with complaints of an 8-year history of facial scarring, swelling of the lower limbs, and a 4-year history of scalp thickening. Physical examination showed thickening furrowing wrinkling of the skin on the face and behind the ears, ciliary body hirsutism, blepharoptosis, and cutis verticis gyrate. Both lower limbs were swollen, especially the knees and ankles. The skin of the palms and soles of the feet was keratinized and thickened. Laboratory examination using bone and joint X-ray showed periostosis of the proximal middle phalanges and metacarpals of both hands, distal ulna and radius, tibia and fibula, distal femurs, and metatarsals.Genetic testing revealed two variants in
8.Research on BP Neural Network Method for Identifying Cell Suspension Concentration Based on GHz Electrochemical Impedance Spectroscopy
An ZHANG ; A-Long TAO ; Qi-Hang RAN ; Xia-Yi LIU ; Zhi-Long WANG ; Bo SUN ; Jia-Feng YAO ; Tong ZHAO
Progress in Biochemistry and Biophysics 2025;52(5):1302-1312
ObjectiveThe rapid advancement of bioanalytical technologies has heightened the demand for high-throughput, label-free, and real-time cellular analysis. Electrochemical impedance spectroscopy (EIS) operating in the GHz frequency range (GHz-EIS) has emerged as a promising tool for characterizing cell suspensions due to its ability to rapidly and non-invasively capture the dielectric properties of cells and their microenvironment. Although GHz-EIS enables rapid and label-free detection of cell suspensions, significant challenges remain in interpreting GHz impedance data for complex samples, limiting the broader application of this technique in cellular research. To address these challenges, this study presents a novel method that integrates GHz-EIS with deep learning algorithms, aiming to improve the precision of cell suspension concentration identification and quantification. This method provides a more efficient and accurate solution for the analysis of GHz impedance data. MethodsThe proposed method comprises two key components: dielectric property dataset construction and backpropagation (BP) neural network modeling. Yeast cell suspensions at varying concentrations were prepared and separately introduced into a coaxial sensor for impedance measurement. The dielectric properties of these suspensions were extracted using a GHz-EIS dielectric property extraction method applied to the measured impedance data. A dielectric properties dataset incorporating concentration labels was subsequently established and divided into training and testing subsets. A BP neural network model employing specific activation functions (ReLU and Leaky ReLU) was then designed. The model was trained and tested using the constructed dataset, and optimal model parameters were obtained through this process. This BP neural network enables automated extraction and analytical processing of dielectric properties, facilitating precise recognition of cell suspension concentrations through data-driven training. ResultsThrough comparative analysis with conventional centrifugal methods, the recognized concentration values of cell suspensions showed high consistency, with relative errors consistently below 5%. Notably, high-concentration samples exhibited even smaller deviations, further validating the precision and reliability of the proposed methodology. To benchmark the recognition performance against different algorithms, two typical approaches—support vector machines (SVM) and K-nearest neighbor (KNN)—were selected for comparison. The proposed method demonstrated superior performance in quantifying cell concentrations. Specifically, the BP neural network achieved a mean absolute percentage error (MAPE) of 2.06% and an R² value of 0.997 across the entire concentration range, demonstrating both high predictive accuracy and excellent model fit. ConclusionThis study demonstrates that the proposed method enables accurate and rapid determination of unknown sample concentrations. By combining GHz-EIS with BP neural network algorithms, efficient identification of cell concentrations is achieved, laying the foundation for the development of a convenient online cell analysis platform and showing significant application prospects. Compared to typical recognition approaches, the proposed method exhibits superior capabilities in recognizing cell suspension concentrations. Furthermore, this methodology not only accelerates research in cell biology and precision medicine but also paves the way for future EIS biosensors capable of intelligent, adaptive analysis in dynamic biological research.
9.Shaoyaotang Restores Th17/Treg Cell Balance by Regulating Glucose Metabolism Reprogramming in Treatment of Ulcerative Colitis
Yiwen WANG ; Yiling XIA ; Erle LIU ; Shaijin JIANG ; Bo ZOU ; Dongsheng WU ; Youwei XIAO ; Hui CAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):78-85
ObjectiveTo investigate the effect of Shaoyaotang on T helper cell 17/regulatory T lymphocyte(Th17/Treg) cell balance in ulcerative colitis and decipher the intervention mechanism based on glucose metabolism reprogramming. MethodsThe mouse model of ulcerative colitis was established by the dextran sulfate sodium (DSS) method. Forty-eight C57BL/6 mice were randomly allocated into normal, model, Western drug control (mesalazine, 0.39 g·kg-1·d-1), Shaoyaotang (15.54 g·kg-1·d-1), inhibitor (2-deoxy-D-glucose, 2-DG, 100 mg·kg-1·d-1), and inhibitor (2-DG, 100 mg·kg-1·d-1) + Shaoyaotang (15.54 g·kg-1·d-1) groups. Mice were administrated with the corresponding drugs by gavage for 7 days. The general conditions and the colon injury degree were observed 24 h after the last administration. The expression of interleukin (IL)-10 and IL-17 in the colon tissue was detected by immunohistochemical staining. Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were performed to determine the protein and mRNA levels, respectively, of hypoxia-inducing factor-1α (HIF-1α), lactate dehydrogenase (LDHA), and hexokinase 2 (HK2) in the colon tissue. Th17/Treg cell differentiation was detected by flow cytometry. Enzyme-linked immunosorbent assay was employed to measure the levels of lactic acid and glucose in the colon tissue and IL-10, IL-17, and IL-6 in the serum. ResultsCompared with the normal group, the model group showed decreases in body weight and disease activity index (DAI) (P<0.05), elevations in levels of HIF-1α, LDHA, HK2, IL-17, IL-6, Th17 cells, lactic acid, and glucose in the colon tissue (P<0.05), and declines in the levels of of IL-10 and Treg cells (P<0.05). Compared with the model group, the drug administration groups showed increases in body weight and DAI (P<0.05), declines in levels of HIF-1α, LDHA, HK2, IL-17, IL-6, Th17 cells, lactic acid, and glucose in the colon tissue (P<0.05), and rises in levels of IL-10 and Treg cells (P<0.05). Shaoyaotang+2-DG group had the most obvious effect. ConclusionShaoyaotang can relieve diarrhea and bloody stool in mice with ulcerative colitis by restoring the Th17/Treg cell balance via regulation of glucose metabolism reprogramming, thus playing a role in the treatment of ulcerative colitis.
10.Shaoyaotang Regulates Glucose Metabolism Reprogramming to Inhibit Macrophage Polarization Toward M1 Phenotype
Shaijin JIANG ; Hui CAO ; Dongsheng WU ; Bo ZOU ; Yiwen WANG ; Yiling XIA ; Erle LIU ; Qi CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):86-93
ObjectiveTo explore the regulation of Shaoyaotang on glucose metabolism reprogramming of macrophages and the mechanism of this decoction in inhibiting macrophage polarization toward the M1 phenotype. MethodsHuman monocytic leukemia-1 (THP-1) cells were treated with 100 ng·L-1 phorbol myristate acetate for induction of macrophages as the normal control group. The cells treated with 100 ng·L-1 lipopolysaccharide combined with 20 ng·L-1 interferon (IFN)-γ for induction of M1-type macrophages were taken as the M1 model group. M1-type macrophages were treated with the blank serum, Shaoyaotang-containing serum, 0.5 mol·L-1 2-deoxy-D-glucose (2-DG), and Shaoyaotang-containing serum + 2-DG, respectively. After intervention, the expression of CD86 and CD206 was examined by flow cytometry. The levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-10, and transforming growth factor (TGF)-β were assessed by ELISA. Real-time PCR and Western blot were employed to determine the mRNA and protein levels, respectively, of hypoxia-inducible factor-1 alpha (HIF-1α), glucose transporter 1 (GLUT1), hexokinase 2 (HK2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3). ResultsCompared with that in the normal control group, the expression of CD86, the marker of M1-type macrophages, increased in the M1 model group and blank serum group (P<0.01), which indicated that the M1 inflammatory model was established successfully. In addition, the M1 model group was observed with up-regulated mRNA and protein levels of proinflammatory cytokines IL-6 and TNF-α and glycolysis-related factors HIF-1α, GLUT1, HK2, GAPDH, and PFKFB3 (P<0.01). Compared with the M1 model group, the Shaoyaotang-containing serum, 2-DG, and combined intervention groups showed decreased expression of CD86 (P<0.01), down-regulated mRNA and protein levels of proinflammatory factors IL-6 and TNF-α and glycolysis-related factors HIF-1α, GLUT1, HK2, GAPDH, and PFKFB3 produced by M1-type macrophages (P<0.01), increased expression of CD206 (marker of M2-type macrophages) (P<0.01), and elevated levels of IL-10 and TGF-β produced by M2-type macrophages (P<0.01). ConclusionShaoyaotang inhibits macrophage differentiation toward pro-inflammatory M1-type macrophages and promotes the differentiation toward anti-inflammatory M2-type macrophages by regulating glucose metabolism reprogramming. The evidence gives insights into new molecular mechanisms and targets for the treatment of ulcerative colitis with Shaoyaotang.

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