1.Mechanism of Paeoniae Radix Rubra and Aconiti Lateralis Radix Praeparata in Treatment of Acute-on-chronic Liver Failure Based on Bioinformation Analysis and Experimental Validation
Xiaoling TIAN ; Yu ZHANG ; Shan DU ; Mengsi WU ; Nianhua TAN ; Bin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):156-165
		                        		
		                        			
		                        			ObjectiveTo explore the mechanism of action of Paeoniae Radix Rubra and Aconiti Lateralis Radix Praeparata (CSFZ) in the treatment of acute-on-chronic liver failure (ACLF) through network pharmacology, molecular docking, and animal experiments. MethodsNetwork pharmacology was used to identify potential targets and related signaling pathways for the treatment of ACLF with CSFZ. Molecular docking was used to examine the binding activity of the core components with corresponding key targets. An ACLF rat model was established by subcutaneous and tail vein injections of bovine serum albumin combined with lipopolysaccharide (LPS) + D-galactosamine (D-GalN) intraperitoneal injection. A normal control group (NC), a model group, a CSFZ group (CSFZ, 5.85 g·kg-1), and a hepatocyte growth-promoting granule group (HGFG, 4.05 g·kg-1) were set up in this study. Pathological changes in rat liver tissue were observed using hematoxylin and eosin (HE) and Masson staining. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of interleukin-6 (IL-6), B-cell lymphoma-2 (Bcl-2), Caspase-3, and albumin (ALB). Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure the mRNA and protein expression levels of phosphoinositide 3-kinase (PI3K), protein kinase B (Akt), phosphorylated PI3K (p-PI3K), and phosphorylated Akt (p-Akt). ResultsNetwork pharmacology screening identified 49 active ingredients of CSFZ, 103 action targets, and 3 317 targets related to ACLF. Among these, 74 targets overlapped with CSFZ drug targets. Key nodes in the protein-protein interaction (PPI) network included Akt1, tumor necrosis factor (TNF), IL-6, Bcl-2, and Caspase-3. Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis identified multiple signaling pathways, with the PI3K/Akt signaling pathway being the most frequent. Molecular docking showed that the core components of the drug exhibited good binding activity with the corresponding key targets. Animal experiments confirmed that CSFZ significantly improved liver tissue pathological damage in ACLF rats, reduced the release of inflammatory factors and liver cell apoptosis, and upregulated the expression levels of the PI3K/Akt signaling pathway. ConclusionThrough network pharmacology, molecular docking, and in vivo experiments, this study confirms the effect of CSFZ in reducing liver cell inflammatory damage and inhibiting liver cell apoptosis. The specific mechanism may be related to its involvement in regulating the PI3K/Akt signaling pathway. 
		                        		
		                        		
		                        		
		                        	
2.Application of deep learning in oral imaging analysis
Yuxuan YANG ; Jingyi TAN ; Lili ZHOU ; Zirui BIAN ; Yifan CHEN ; Yanmin WU
Chinese Journal of Tissue Engineering Research 2025;29(11):2385-2393
		                        		
		                        			
		                        			BACKGROUND:In recent years,deep learning technologies have been increasingly applied in the field of oral medicine,enhancing the efficiency and accuracy of oral imaging analysis and promoting the rapid development of intelligent oral medicine. OBJECTIVE:To elaborate the current research status,advantages,and limitations of deep learning based on oral imaging in the diagnosis and treatment decision-making of oral diseases,as well as future prospects,exploring new directions for the transformation of oral medicine under the backdrop of deep learning technology. METHODS:PubMed was searched for literature related to deep learning in oral medical imaging published from January 2017 to January 2024 with the search terms"deep learning,artificial intelligence,stomatology,oral medical imaging."According to the inclusion criteria,80 papers were finally included for review. RESULTS AND CONCLUSION:(1)Classic deep learning models include artificial neural networks,convolutional neural networks,recurrent neural networks,and generative adversarial networks.Scholars have used these models in competitive or cooperative forms to achieve more efficient interpretation of oral medical images.(2)In the field of oral medicine,the diagnosis of diseases and the formulation of treatment plans largely depend on the interpretation of medical imaging data.Deep learning technology,with its strong image processing capabilities,aids in the diagnosis of diseases such as dental caries,periapical periodontitis,vertical root fractures,periodontal disease,and jaw cysts,as well as preoperative assessments for procedures such as third molar extraction and cervical lymph node dissection,helping clinicians improve the accuracy and efficiency of decision-making.(3)Although deep learning is promising as an important auxiliary tool for the diagnosis and treatment of oral diseases,it still has certain limitations in model technology,safety ethics,and legal regulation.Future research should focus on demonstrating the scalability,robustness,and clinical practicality of deep learning,and finding the best way to integrate automated deep learning decision support systems into routine clinical workflows.
		                        		
		                        		
		                        		
		                        	
3.Treadmill training activates endogenous neural stem cells to promote spinal cord injury repair in mice
Chanjuan CHEN ; Zeyu SHANGGUAN ; Qizhe LI ; Wei TAN ; Qing LI
Chinese Journal of Tissue Engineering Research 2025;29(19):3976-3982
		                        		
		                        			
		                        			BACKGROUND:Treadmill training is one of the effective ways to promote the recovery of motor function after spinal cord injury.Treadmill training can promote neurogenesis,but the effect of different intensities of treadmill training on the activation of endogenous stem cells is still unclear. OBJECTIVE:To analyze the activation effect of different intensities of treadmill training on endogenous neural stem cells in the spinal cord of mice after spinal cord injury. METHODS:Fifty female C57BL/6J mice were divided into control group,spinal cord injury group,low-,moderate-,and high-intensity exercise groups with 10 mice in each group by random number table method.T10 segment spinal cord injury model was constructed by the clamp method in spinal cord injury group,low-,moderate-,and high-intensity exercise groups.On day 7 after spinal cord injury,mice in the low-,moderate-,and high-intensity exercise groups were respectively trained on the treadmill with corresponding intensity,3 times/d,10 min/times,6 times a week for 28 consecutive days.At 3,7,14,21,and 28 days after treadmill training,the hind limb motor function was evaluated by BMS score.At 28 days after treadmill training,the spinal cord tissue of the injured area was obtained,and the expression of epidermal growth factor receptor,glial fibrillary acidic protein,and 5-Ethynyl-2'-deoxyuridine(EdU),a proliferative marker,was detected.Hematoxylin-eosin staining was used to observe the morphology of spinal cord. RESULTS AND CONCLUSION:(1)The BMS score of mice in the spinal cord injury group was lower than that in the control group(P<0.05).With the extension of treadmill training time,the BMS scores of mice with spinal cord injury gradually increased,and the BMS scores of mice in moderate-intensity exercise group on days 14 and 21 after treadmill training were higher than those in spinal cord injury group and low-and high-intensity exercise groups(P<0.05).The BMS score of mice in moderate-and high-intensity exercise group was higher than that in spinal cord injury group and low-intensity exercise group at 28 days after treadmill training(P<0.05).(2)Compared with the control group,the proportion of epidermal growth factor receptor and EdU positive cells was increased in spinal cord injury group(P<0.05).Compared with spinal cord injury group,the proportion of epidermal growth factor receptor and EdU positive cells was increased in low-,moderate-,and high-intensity exercise groups(P<0.05),and the highest was found in moderate-intensity exercise group.Compared with control group,the proportion of glial fibrillary acidic protein positive cells was increased in spinal cord injury group(P<0.05).Compared with spinal cord injury group,the proportion of glial fibrillary acidic protein positive cells was lower in low-,moderate-,and high-intensity exercise groups(P<0.05),and the moderate-intensity exercise group was the lowest.(3)Hematoxylin-eosin staining showed that a large cavity was formed in the injured area of mice with spinal cord injury,and the cavity in the injured area of mice with spinal cord injury decreased after different intensities of treadmill training,and the decrease was most obvious in the moderate-intensity exercise group.(4)These results indicate that low-,moderate-,and high-intensity treadmill training can promote the recovery of motor function of mice with spinal cord injury by activating endogenous neural stem cells,and the effect of moderate-intensity exercise training is the most obvious.
		                        		
		                        		
		                        		
		                        	
4.Study on the influential factors for clinical efficacy of polymyxin B combined with other antibiotics in the treat-ment of carbapenem-resistant Acinetobacter baumannii pulmonary infection
Mei DU ; Ruijuan TAN ; Lidan WANG ; Hejun CHEN ; Hanze LI ; Yuanyuan ZHANG
China Pharmacy 2025;36(12):1495-1499
		                        		
		                        			
		                        			OBJECTIVE To analyze the influential factors for clinical efficacy of polymyxin B combined with other antibiotics in the treatment of carbapenem-resistant Acinetobacter baumannii (CRAB) pulmonary infection. METHODS A retrospective analysis was conducted on the clinical data of patients with CRAB pulmonary infection in our hospital from May 2021 to October 2024. Information such as age, gender, admitting department, infection status, underlying medical conditions, mechanical ventilation time, combination anti-infective treatment regimens, and the Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE-Ⅱ) score 24 h before medication was compiled. Based on the effectiveness of the treatment, patients were divided into treatment-effective group and treatment-ineffective group. Univariate analysis and multivariate Logistic regression analysis were employed to identify independent factors influencing clinical efficacy. RESULTS A total of 156 patients were included, and 108 patients were treated effectively, with an effective rate of 69.23%. The results of univariate analysis indicated that there were statistically significant differences between 2 groups in terms of the duration of mechanical ventilation time, APACHE-Ⅱ score 24 h before medication, the number of complication types, the proportion of abnormal coagulation function, anti-infective treatment course, and hospital stay before medication (P<0.05). The results of multivariate Logistic regression analysis showed that APACHE-Ⅱ score≥15 points 24 h before medication [OR=2.965, 95%CI (1.284, 6.845), P=0.020], mechanical 20251606) ventilation time≥10 d [OR=3.577, 95%CI (1.185, 10.793), P=0.037] and hospital stay≥14 d before medication [OR=2.422, 95%CI (1.036, 5.654), P=0.041] were independent 15120420253@139.com risk factors, and anti-infective treatment course>7 d was a protective factor [OR=0.445, 95%CI (0.221, 0.895), P=0.043]. CONCLUSIONS This study shows that the effective rate of polymyxin B combined with other antibiotics in the treatment of CRAB pulmonary infection is less than 70%. The mechanical ventilation time≥10 d, APACHE-Ⅱ score≥15 points 24 h before medication, and hospital stay≥14 d before medication may lead to treatment failure, whereas anti-infective treatment course>7 d may be associated with treatment success.
		                        		
		                        		
		                        		
		                        	
5.Polygonatum Sibiricum Polysaccharides Improve Colonic Injury in a Mouse Model of Chronic Obstructive Pulmonary Disease by Regulating Bile Acid Metabolism in the Colon
Wanrong LI ; Mengting TAO ; Yuanfeng ZOU ; Dan HE ; Nengyuan TANG ; Xin TAN ; Lixia LI ; Dandan CHEN
Journal of Sun Yat-sen University(Medical Sciences) 2025;46(3):431-443
		                        		
		                        			
		                        			ObjectiveTo investigate the effect and mechanism of Polygonatum neutral polysaccharides from sibiricum (PSP-NP) on colon injury in mice with chronic obstructive pulmonary disease (COPD). MethodsMale C57BL/6J mice were randomly divided into a control group, a COPD model group, and a PSP-NP group. The COPD model was established using smoke exposure combined with intranasal LPS administration. The PSP-NP group was simultaneously treated daily with 200 mg/kg of PSP-NP via intragastric gavage, while the other groups received an equal volume of saline. HE staining was used to observe the pathological changes in the colon. ELISA was employed to detect the levels of LPS in serum and the expressions of ZO-1, Occludin, IL-6, and TNF-α in colon tissue. UPLC-MS was used to detect the types and contents of bile acids in colonic content, and to screen for differential bile acids. Differential microbial flora were identified using 16S rRNA gene sequencing, and correlation analysis was conducted with differential bile acids. PSP-NP was combined with the differential bile acids cholic acid (CA), and deoxycholic acid (DCA) in vitro to analyze the binding capacity of PSP-NP for CA and DCA. PSP-NP was applied to NCM460 normal colonic epithelial cells cultured in CA and DCA. Cell migration ability was assessed using the scratch assay, and the mRNA expression levels of inflammatory cytokines TNF-α, IL-6, and NF-κB were measured by RT-qPCR. ResultsPSP-NP effectively improved colonic damage in COPD model mice, enhanced mechanical barrier function, alleviated inflammatory response, and regulated abnormal changes in colonic flora and bile acid metabolism. Correlation analysis further revealed that PSP-NP regulated colonic bile acid metabolism and reduced the redundancy of secondary bile acids by increasing the relative abundance of Bacteroidota, Verrucomicrobiota, Bacteroides, and Akkermansia, while decreasing the relative abundance of Lactobacillus and Bifidobacterium. Notably, in vitro binding assays demonstrated that PSP-NP bound to differential bile acids DCA and CA, with the strongest binding capacity for DCA at 58.2%. In cellular functional studies, DCA inhibited the migration ability of colonic epithelial cells NCM460 and significantly increased the relative mRNA expression levels of inflammatory factors TNF-α, IL-6, and NF-κB. Importantly, co-treatment with PSP-NP significantly ameliorated the impact of DCA on NCM460 cells. ConclusionsPSP-NP may significantly improve colonic damage in COPD model mice. The mechanism may involve the regulation of colonic bile acid metabolism and bile acid profiles through both microbial modulation and direct binding, thereby reducing the damage caused by secondary bile acids such as DCA to colonic epithelial cells. 
		                        		
		                        		
		                        		
		                        	
6.Relationship between gaming motivation and gaming disorder symptoms among adolescents: a network analysis approach
Xinjie TAN ; Shuanghong CHEN ; Chunlin LIU ; Wanjun YANG ; Ying HE
Sichuan Mental Health 2025;38(3):261-266
		                        		
		                        			
		                        			BackgroundPreventing and intervening in adolescent gaming disorder is of significant practical importance. Gaming motivation is strongly linked to gaming addiction and serves a key function in comprehending and addressing addictive gaming behaviors. However, the relationship between components of gaming motivation and symptoms of gaming disorder remains unclear. ObjectiveTo explore the relationship between components of gaming motivation and symptoms of gaming disorder among adolescents, so as to provide references for the prevention and intervention of gaming disorder in this population. MethodsFrom January to February 2024, a cluster sampling method was employed to select 1 414 adolescents from four middle schools in Sichuan Province and Chongqing Municipality as participants in the study. Online Game Motivation Scale (OGMS) and Gaming Disorder Scale for Adolescents (GADIS-A) were administered. Network analysis methods were utilized to investigate the relationships between components of gaming motivation and symptoms of gaming disorder. ResultsThe network edge weights revealed that achievement motivation was positively correlated with impaired game control ability, continued gaming despite negative consequences and the frequency of symptom occurrence (r=0.115, 0.050, 0.076, P<0.05). Social motivation was negatively correlated with negative consequences (r=-0.054, P<0.05),while immersion motivation was positively correlated with continued gaming despite negative consequences (r=0.032, P<0.05). Achievement motivation exhibited the highest strength centrality (1.157) among the three components of gaming motivation. ConclusionThe connections between components of gaming motivation and symptoms of gaming disorder exhibit distinct patterns, with each motivational component influencing gaming disorder through specific symptom pathway. Among these components, achievement motivation plays the most critical role in the interplay between gaming motivation and symptoms of gaming disorder. [Funded by Chongqing Science and Health Joint Medical Science and Technology Innovation Projects General Projects (number, 2023MSXM133)] 
		                        		
		                        		
		                        		
		                        	
7.Effect of Yinchenhao Tang Combined with Yinchen Zhufu Tang on Treg/Th17 Cells in Vitro from Patients with Hepatitis B Virus-related Acute-on-chronic Liver Failure
Menghui ZENG ; Shan DU ; Nianhua TAN ; Jie PENG ; Bin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):43-51
		                        		
		                        			
		                        			ObjectiveTo study the regulatory effects of Yinchenhao Tang combined with Yinchen Zhufu Tang on the expression of regulatory T (Treg)/helper T 17 (Th17) cells cultured in vitro from the patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF). MethodsFresh peripheral blood was collected from the patients with HBV-ACLF for the separation of peripheral blood mononuclear cells (PBMCs). Immunomagnetic beads were used to isolate primary Treg and naive CD4+ T cells. After in vitro expansion, naive CD4+ T cells were induced to differentiate into Th17 cells. Rats were treated with the clearing method (Yinchenhao Tang), warming method (Yinchen Zhufu Tang), and combination of clearing method with warming method (Yinchenhao Tang combined with Yinchen Zhufu Tang, also known as Wenyang Jiedu Huayu Prescription), respectively, and then the medicated plasma samples were collected. Meanwhile, blank plasma was collected from the rats treated with normal saline. Cells were classified into blank, clearing method (5.04 g·kg-1), warming method (6.21 g·kg-1), and combination of clearing method with warming method (17.1 g·kg-1) groups and treated with corresponding plasma. The frequency of Treg/Th17 cells was detected by flow cytometry. The level of transforming growth factor-β (TGF-β) was measured by the enzyme-linked immunosorbent assay. The cytometric bead array (CBA) was employed to measure the levels of interleukin-10 (IL-10), interleukin-17A (IL-17A), tumor necrosis factor-α (TNF-α), and interleukin-23 (IL-23). The mRNA and protein levels of Forkhead box P3 (FoxP3) and retinoic acid-related orphan receptor-gamma t (ROR-γt) were determined by Real-time PCR and Western blot, respectively. ResultsCompared with the blank group, the combination of clearing method with warming method group showed decreased frequency of Treg and Th17 cells, lowered levels of Treg cytokines (TGF-β and IL-10) and Th17 cytokines (TNF-α, IL-17A, and IL-23), and down-regulated mRNA and protein levels of FoxP3 and ROR-γt (P<0.01). Compared with the clearing method group, the combination of clearing method with warming method group showed decreased Treg cell frequency and down-regulated mRNA and protein levels of FoxP3. Meanwhile, the combination group showed decreased Th17 cell frequency, lowered levels of TGF-β, IL-10, TNF-α, IL-17A, and IL-23, and down-regulated mRNA and protein levels of ROR-γt (P<0.05, P<0.01). Compared with the warming method group, the combination of clearing method with warming method group showed decreased frequency of Treg cells and down-regulated mRNA and protein levels of FoxP3. Meanwhile, the combination group showed decreased Th17 cell frequency, declined levels of TGF-β, IL-10, TNF-α, IL-17A, and IL-23, and down-regulated mRNA and protein levels of ROR-γt (P<0.05). ConclusionThe combination of clearing method with warming method can down-regulate the expression of specific cytokines of Treg and Th17 cells, inhibit the over activation of Treg and Th17 cells, and reduce the secretion of cytokines such as TGF-β, IL-10, TNF-α, IL-17A, and IL-23, thereby alleviating inflammation and improving the prognosis of the patients with liver failure. 
		                        		
		                        		
		                        		
		                        	
8.Mechanism of Xiayuxue Tang in Inhibiting Hepatocellular Carcinoma Cells by Regulating YAP1/SIRT5 Signaling Axis to Mediate Succinate Metabolism and Succinylation
Linzhu LU ; Qianqian GUO ; Xuefei TIAN ; Bin CHEN ; Nianhua TAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):52-61
		                        		
		                        			
		                        			ObjectiveTo investigate the inhibitory effects and mechanisms of Xiayuxue Tang (XYXT) on hepatocellular carcinoma cells through the regulation of succinate metabolism and succinylation modification. MethodsXYXT-medicated serum was prepared. The effects of different concentrations of succinate on the proliferation of HepG2 and MHCC97H cells were observed using the cell counting kit-8 (CCK-8) assay, and the experimental concentrations for subsequent tests were determined. Further CCK-8 assays were performed to evaluate the effects of XYXT-medicated serum of different concentrations (5%, 10%, and 15%) on cell proliferation. Flow cytometry, scratch test, and Transwell assay were employed to analyze the effect of 10% XYXT-medicated serum on the cell cycle, migration, invasion, and apoptosis. The changes in succinate metabolism and succinylation modification were examined based on succinate content assays, succinate dehydrogenase (SDH) activity detection, and western blot. The expressions of Yes-associated protein 1 (YAP1) and sirtuin 5 (SIRT5) were detected via Real-time PCR and western blot. Molecular docking was applied to validate the binding between XYXT’s main components and target proteins. ResultsCompared with the control group, 1-2 mmol·L-¹ succinate significantly promoted HepG2 and MHCC97H cell proliferation (P<0.01). XYXT-medicated serum (5%, 10%, and 15%) markedly inhibited the proliferation of both cell lines compared with the blank group (P<0.05, P<0.01). Treatment with 10% XYXT-medicated serum arrested the cell cycle at the stage prior to DNA synthesis (G0/G1) (P<0.01), suppressed migration (P<0.01) and invasion (P<0.05, P<0.01), and promoted apoptosis of HepG2 and MHCC97H cells (P<0.01). Co-treatment with XYXT and succinate reversed the inhibitory effects of XYXT on proliferation, migration, and invasion of HepG2 and MHCC97H cells (P<0.05, P<0.01). The proportion of cells at G0/G1 phase decreased (P<0.01), and the apoptosis rate decreased (P<0.01). In terms of succinate metabolism, compared with the blank serum group, the 10% XYXT-medicated serum reduced succinate levels of HepG2 and MHCC97H cells (P<0.05, P<0.01), enhanced SDH activity (P<0.01), and downregulated succinylation modification. In terms of YAP1/SIRT5 pathway, compared with the blank serum group, the 10% XYXT-medicated serum significantly downregulated the mRNA and protein expressions of YAP1 in HepG2 and MHCC97H cells (P<0.05, P<0.01) while significantly upregulated the mRNA and protein expressions of SIRT5 (P<0.05, P<0.01). Molecular docking confirmed that there was a good binding ability between YAP1 and SIRT5, as well as between XYXT's main active components and YAP1 and SIRT5. ConclusionXYXT suppresses hepatocellular carcinoma cells by modulating the YAP1/SIRT5 signaling axis to intervene in succinate metabolism and succinylation modification. 
		                        		
		                        		
		                        		
		                        	
9.Differences in Expression of Intestinal Flora in Patients with Different TCM Syndrome Types of "Yang Huang-Yin-Yang Huang-Yin Huang" in Hepatitis B Virus-related Acute-on-chronic Liver Failure
Shan DU ; Menghui ZENG ; Nianhua TAN ; Jie PENG ; Bin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):62-69
		                        		
		                        			
		                        			ObjectiveTo investigate the differential expression of intestinal flora in patients with different traditional Chinese medicine (TCM) syndrome types (Yang Huang syndrome, Yin-Yang Huang syndrome, and Yin Huang syndrome) of hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) and clarify the biological basis of jaundice and Yin Huang syndrome in liver failure. MethodsA total of 20 cases of HBV-ACLF patients were included in the Yang Huang group, 20 cases in the Yin-Yang Huang group, 16 cases in the Yin Huang group, and 20 healthy adult volunteers. 16S rRNA gene sequencing was used to detect the diversity, species distribution, and differences of the subjects' intestinal flora, and bioinformatics analysis was conducted. ResultsCompared with those in the healthy control group, the species richness and diversity of intestinal flora in the HBV-ACLF Yang Huang group, Yin-Yang Huang group, and Yin Huang group were significantly reduced, and there were significant differences in the composition of intestinal flora compared with healthy volunteers. However, there were no significant differences in the species richness, diversity, and composition of intestinal flora among the three groups. LEfSe analysis showed that compared with the healthy control group, the HBV-ACLF Yang Huang group showed significant enrichment of Staphylococcus aureus(P<0.01). Yin-Yang Huang group showed significant enrichment of s_Ileibacterium valens(P<0.05,P<0.01), and the Yin Huang group showed significant enrichment of Enterococcus faecium and Streptococcus sali varius(P<0.05). These strains may be biomarkers between the three groups of patients and the healthy control group. Compared with that in the Yin-Yang Huang group, Tyzzerella_nexilis was significantly enriched in the Yang-Huang group, and Streptococcus lactiae was significantly enriched in the Yin-Yang Huang group. Compared with that in the Yang-Huang group and the Yin-Yang Huang group, Enterococcus faecalis was significantly enriched in the Yin Huang group. The above strains may be biomarkers among the three groups of patients, and Enterococcus faecium may be a biomarker for the transition from the Yang Huang group to the Yin Huang group. ConclusionsThere are significant differences in the intestinal flora between patients with HBV-ACLF Yang Huang syndrome, Yin-Yang Huang syndrome, and Yin Huang syndrome. Enterococcus faecium is significantly enriched in the Yin Huang syndrome group, suggesting that dysbiosis of the intestinal flora may be the biological basis for jaundice and Yin Huang syndrome in liver failure. 
		                        		
		                        		
		                        		
		                        	
10.Changes of Treg/Th17 Cell Expression in Peripheral Blood of Patients with Hepatitis B Virus-related Acute-on-chronic Liver Failure in Five Types of Traditional Chinese Medicine Syndrome
Shan DU ; Menghui ZENG ; Nianhua TAN ; Jie PENG ; Bin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):70-76
		                        		
		                        			
		                        			ObjectiveTo study the expression differences of regulatory T cells (Treg) and T helper 17 cells (Th17) in the peripheral blood of patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) in five types of traditional Chinese medicine (TCM) syndrome. MethodsA total of 144 patients with HBV-ACLF were included and divided into five types of TCM syndrome, including 34 cases of heat-toxin amassment syndrome, 44 cases of dampness-heat amassment syndrome, 27 cases of Qi-deficiency and stasis jaundice syndrome, 21 cases of spleen-kidney Yang deficiency syndrome, and 18 cases of liver-kidney Yin deficiency syndrome. Meanwhile, 30 healthy volunteers were included as controls. The frequency of Treg and Th17 cells in the peripheral blood of subjects in each group was detected by flow cytometry, and the Treg/Th17 ratio was calculated. Cytometric bead array (CBA) was used to detect the levels of cytokines interleukin (IL)-10, transforming growth factor-β (TGF-β), tumor necrosis factor-α (TNF-α), IL-17A, and IL-23. Real-time fluorescence quantitative PCR (Real-time PCR) detected the mRNA expression of forkhead box P3 (FoxP3) and retinoic acid-related orphan receptor gamma t (ROR-γt). Results(1) Compared with that in the healthy control group, the frequency of Treg and Th17 cells in patients with various TCM syndrome types of HBV-ACLF increased (P<0.05). Compared with that in the heat-toxin amassment syndrome group, the frequency of Treg and Th17 cells decreased in the dampness-heat amassment syndrome group (P<0.05), while the frequency of Treg and Th17 cells increased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the dampness-heat amassment syndrome group, the frequency of Treg and Th17 cells increased in the dampness-heat amassment syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the Qi-deficiency and stasis jaundice syndrome group, the frequency of Treg and Th17 cells increased in the spleen-kidney Yang deficiency syndrome group (P<0.05), while the frequency of Treg and Th17 cells decreased in the liver-kidney Yin deficiency syndrome group (P<0.05). Compared with the spleen-kidney Yang deficiency syndrome group, the frequency of Treg and Th17 cells decreased in the liver-kidney Yin deficiency syndrome group (P<0.05). (2) Compared with that in the healthy control group, the Treg/Th17 cell ratio in patients with various TCM syndromes of HBV-ACLF decreased (P<0.05). Compared with that in the heat-toxin amassment syndrome group, the Treg/Th17 cell ratio increased in the dampness-heat amassment syndrome group (P<0.05), while it decreased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the dampness-heat amassment syndrome group, the Treg/Th17 cell ratio decreased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the Qi-deficiency and stasis jaundice syndrome group, the Treg/Th17 cell ratio increased in the spleen-kidney Yang deficiency syndrome group and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with the spleen-kidney Yang deficiency syndrome group, the Treg/Th17 cell ratio in the liver-kidney Yin deficiency syndrome group increased (P<0.05). (3) Compared with those in the healthy control group, the levels of Treg-related cytokines IL-10 and TGF-β, as well as Th17-related cytokines TNF-α, IL-17A, and IL-23, were elevated in patients with various TCM syndrome types of HBV-ACLF (P<0.05). There was no significant difference in TNF-α levels among different TCM syndrome types. Compared with those in the heat-toxin amassment syndrome group, the levels of IL-10, TNF-β, IL-17A, and IL-23 in the dampness-heat amassment syndrome group, Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome groups increased (P<0.05). Compared with those in the dampness-heat amassment syndrome group, the levels of IL-10, TGF-β, IL-17A, and IL-23 increased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with those in the Qi-deficiency and stasis jaundice syndrome group, the levels of IL-10, TGF-β, IL-17A, and IL-23 in the spleen-kidney Yang deficiency syndrome group increased (P<0.05), while those in the liver-kidney Yin deficiency syndrome group decreased (P<0.05). Compared with those in the spleen-kidney Yang deficiency syndrome, the levels of IL-10, TGF-β, IL-17A, and IL-23 in the liver-kidney Yin deficiency syndrome group decreased (P<0.05). (4) Compared with that in the healthy control group, the mRNA of Treg/Th17 cell specific transcription factors FoxP3 and ROR-γt were elevated in patients with various TCM syndrome types of HBV-ACLF (P<0.05). Compared with that in the heat-toxin amassment syndrome group, the mRNA of FoxP3 and ROR-γt increased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the dampness-heat amassment syndrome group, the mRNA of FoxP3 and ROR-γt increased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome (P<0.05). Compared with that in the Qi-deficiency and stasis jaundice syndrome group, the mRNA of FoxP3 and ROR-γt in the spleen-kidney Yang deficiency syndrome group increased (P<0.05), and it decreased in the liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the spleen-kidney Yang deficiency syndrome group, the mRNA of FoxP3 and ROR-γt decreased in the liver-kidney Yin deficiency syndrome group (P<0.05). ConclusionThe frequency and ratio of Treg/Th17 cells, as well as the expression of related cytokines and specific receptors in peripheral blood of patients with HBV-ACLF in five types of TCM syndromes are different, which has certain reference value for TCM syndrome differentiation and treatment of patients with HBV-ACLF. 
		                        		
		                        		
		                        		
		                        	
            
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