1.Effect of different glaucoma surgeries on postoperative refractive status
International Eye Science 2026;26(3):417-423
Glaucoma, the leading cause of irreversible blindness worldwide, remains a central focus of ophthalmic research, particularly with regard to surgical management. Conventional procedures, such as trabeculectomy with mitomycin and glaucoma drainage device implantation, continue to be considered the gold standard because of their strong intraocular pressure lowering efficacy. However, these operations are associated with relatively high rates of postoperative complications, and perioperative fluctuations in intraocular parameters can introduce refractive prediction errors that ultimately compromise visual quality. In recent years, minimally invasive glaucoma surgery(MIGS)has gained increasing attention for its advantages in reducing complications, shortening operative time, minimizing incision size, and accelerating visual recovery, while better preserving postoperative refractive stability. This review systematically summarizes the differences in refractive outcomes between conventional surgery and MIGS, examines the underlying mechanisms, and discusses practical clinical strategies to manage refractive shifts. The aim is to provide a theoretical foundation for precise refractive management in glaucoma surgery, thereby enhancing patients' visual quality and overall quality of life.
2.Exploring on Processing Mechanism of Enhanced "Invigorating Spleen and Stopping Diarrhea" Effect of Soil-fried Atractylodis Macrocephalae Rhizoma Based on "Microscopic Characterization, Chemical Analysis and Pharmacodynamic Evaluation" Trinity
Guoshun SHAN ; Yuyan XIAO ; Chu YUAN ; Xiuai CHEN ; Qimiao ZHAO ; Xiang LIU ; Hao WU ; Ke ZHANG ; Siqi LIU ; Yongduo YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):182-193
ObjectiveTo analyze the processing mechanism underlying the enhanced effect of invigorating spleen and stopping diarrhea of soil-fried Atractylodis Macrocephalae Rhizoma(AMR) by analyzing the changes of microstructure, chemical composition and anti-ulcerative colitis(UC) activity before and after soil stir-frying. MethodsThe microstructure and elemental composition of AMR before and after soil stir-frying were analyzed by scanning electron microscopy-energy dispersive spectroscopy(SEM-EDS), to investigate the differences in microstructure and the underlying causes. Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) coupled with UNIFI 1.9.2 natural product analysis platform were used to analyze and identify the chemical constituents in raw and soil-fried products, and multivariate statistical methods including principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to explore the differences and sources of chemical constituents between them. A dextran sulfate sodium(DSS)-induced UC mouse model was established. The method of disease activity index(DAI) was used to evaluate the severity of intestinal inflammation. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of colon tissue, enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory factors, Real-time quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to analyze the expressions of key genes and proteins involved in the intestinal mucosal barrier. The 16S rRNA sequencing was used to evaluate the diversity of intestinal flora, headspace gas chromatography-mass spectrometry(HS-GC-MS) was used to explore the levels of short-chain fatty acids(SCFAs) in feces. Base on the above findings, this paper investigated the effects of raw and soil-fried AMR on the biological, chemical, mechanical and immune barriers of model animals, and the differences in pharmacological effects and underlying mechanisms from the perspective of regulating the intestinal mucosal barrier in UC mice. ResultsSEM observation revealed numerous hearth soil particles on the surface of soil-fried AMR, accompanied by bubble-like bulges. At the same time, there were many cracks and folds on the surface of the hearth soil. EDS analysis revealed that the contents of Si, Al, Mg and Ca in soil-fried AMR were significantly higher than those of raw products, and these elements constituted the primary components of hearth soil. UPLC-Q-TOF-MS combined with database comparison was used to identify the chemical constituents of raw and soil-fried AMR. In positive ion mode, a total of 132 components were identified, primarily comprising three categories of terpenoids, polyphenols and amino acids. In negative ion mode, a total of 40 components were identified, primarily polyphenolic and glycoside compounds. Among them, the contents of sesquiterpenes and polyphenolic acids were changed significantly before and after processing. Soil-fried AMR could reduce the DAI score of UC mice, alleviate the shortening of colon length, reduce the levels of pro-inflammatory factors such as interleukin(IL)-17, IL-18, γ-interferon(IFN-γ) and tumor necrosis factor(TNF)-α in serum, increase the levels of anti-inflammatory factors such as secretory immunoglobulin A(sIgA), IL-10, IL-4 and transforming growth factor-β(TGF-β) in serum, increase the expressions of key genes and proteins of intestinal mucosal barrier such as tight junction protein-1(ZO-1), Occludin, Claudin-1 and mucin 2(MUC2) in colonic mucosa, and improve the disorders of intestinal flora diversity and the levels of SCFAs(P<0.05, P<0.01). The raw and stir-fried products of AMR also exhibited the aforementioned effects, but they were weaker than the soil-fried products. Additionally, the auxiliary material hearth soil also had a certain pharmacodynamic effect. ConclusionSoil-fried AMR can enhance the protective effect on intestinal mucosal barrier in UC mice. These changes or heating-induced alterations in the microscopic structure and chemical composition of AMR may be attributed to the dual effects of adsorption of hearth soil.
3.Exploring on Processing Mechanism of Enhanced "Invigorating Spleen and Stopping Diarrhea" Effect of Soil-fried Atractylodis Macrocephalae Rhizoma Based on "Microscopic Characterization, Chemical Analysis and Pharmacodynamic Evaluation" Trinity
Guoshun SHAN ; Yuyan XIAO ; Chu YUAN ; Xiuai CHEN ; Qimiao ZHAO ; Xiang LIU ; Hao WU ; Ke ZHANG ; Siqi LIU ; Yongduo YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):182-193
ObjectiveTo analyze the processing mechanism underlying the enhanced effect of invigorating spleen and stopping diarrhea of soil-fried Atractylodis Macrocephalae Rhizoma(AMR) by analyzing the changes of microstructure, chemical composition and anti-ulcerative colitis(UC) activity before and after soil stir-frying. MethodsThe microstructure and elemental composition of AMR before and after soil stir-frying were analyzed by scanning electron microscopy-energy dispersive spectroscopy(SEM-EDS), to investigate the differences in microstructure and the underlying causes. Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) coupled with UNIFI 1.9.2 natural product analysis platform were used to analyze and identify the chemical constituents in raw and soil-fried products, and multivariate statistical methods including principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to explore the differences and sources of chemical constituents between them. A dextran sulfate sodium(DSS)-induced UC mouse model was established. The method of disease activity index(DAI) was used to evaluate the severity of intestinal inflammation. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of colon tissue, enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory factors, Real-time quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to analyze the expressions of key genes and proteins involved in the intestinal mucosal barrier. The 16S rRNA sequencing was used to evaluate the diversity of intestinal flora, headspace gas chromatography-mass spectrometry(HS-GC-MS) was used to explore the levels of short-chain fatty acids(SCFAs) in feces. Base on the above findings, this paper investigated the effects of raw and soil-fried AMR on the biological, chemical, mechanical and immune barriers of model animals, and the differences in pharmacological effects and underlying mechanisms from the perspective of regulating the intestinal mucosal barrier in UC mice. ResultsSEM observation revealed numerous hearth soil particles on the surface of soil-fried AMR, accompanied by bubble-like bulges. At the same time, there were many cracks and folds on the surface of the hearth soil. EDS analysis revealed that the contents of Si, Al, Mg and Ca in soil-fried AMR were significantly higher than those of raw products, and these elements constituted the primary components of hearth soil. UPLC-Q-TOF-MS combined with database comparison was used to identify the chemical constituents of raw and soil-fried AMR. In positive ion mode, a total of 132 components were identified, primarily comprising three categories of terpenoids, polyphenols and amino acids. In negative ion mode, a total of 40 components were identified, primarily polyphenolic and glycoside compounds. Among them, the contents of sesquiterpenes and polyphenolic acids were changed significantly before and after processing. Soil-fried AMR could reduce the DAI score of UC mice, alleviate the shortening of colon length, reduce the levels of pro-inflammatory factors such as interleukin(IL)-17, IL-18, γ-interferon(IFN-γ) and tumor necrosis factor(TNF)-α in serum, increase the levels of anti-inflammatory factors such as secretory immunoglobulin A(sIgA), IL-10, IL-4 and transforming growth factor-β(TGF-β) in serum, increase the expressions of key genes and proteins of intestinal mucosal barrier such as tight junction protein-1(ZO-1), Occludin, Claudin-1 and mucin 2(MUC2) in colonic mucosa, and improve the disorders of intestinal flora diversity and the levels of SCFAs(P<0.05, P<0.01). The raw and stir-fried products of AMR also exhibited the aforementioned effects, but they were weaker than the soil-fried products. Additionally, the auxiliary material hearth soil also had a certain pharmacodynamic effect. ConclusionSoil-fried AMR can enhance the protective effect on intestinal mucosal barrier in UC mice. These changes or heating-induced alterations in the microscopic structure and chemical composition of AMR may be attributed to the dual effects of adsorption of hearth soil.
4.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.
5.Effect of Modified Shibaotang on Serum Sex Hormone Levels in Patients with Male Late-onset Hypogonadism of Kidney Essence Deficiency Syndrome Complicated with Diabetes Mellitus
Yi SHAN ; Shaokang CHEN ; Zhenfu SHI ; Haifeng XU ; Yi LU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):227-233
ObjectiveTo observe the effect of modified Shibaotang on serum sex hormone levels in patients with male late-onset hypogonadism of kidney essence deficiency syndrome complicated with diabetes mellitus. MethodsA total of 60 patients with male late-onset hypogonadism of kidney essence deficiency syndrome complicated with diabetes mellitus,who met the inclusion criteria and were admitted to Yinchuan Hospital of Traditional Chinese Medicine from October 2022 to October 2023,were selected and randomly divided into an observation group and a control group,with 30 patients in each group. Both groups continued their original treatments,including blood glucose lowering and blood lipid regulation. The observation group was treated with modified Shibaotang,while the control group was treated with testosterone undecanoate capsules. The treatment lasted for 12 weeks. The changes in traditional Chinese medicine (TCM)syndrome scores,partial androgen deficiency in aging males (PADAM)symptom scores,glucose metabolism indexes [fasting plasma glucose (FPG),2-hour postprandial glucose (2 h PG),glycosylated hemoglobin (HbA1c)],and serum sex hormone indexes [sex hormone-binding globulin (SHBG),free testosterone (FT),total testosterone (TT),prolactin (PRL),luteinizing hormone (LH),follicle stimulating hormone (FSH),estrogen (E2)] were compared between the two groups before and after treatment. Safety was also evaluated. Results(1)Clinical efficacy comparison:After treatment,the clinical efficacy in both groups was similar,and there was no statistically significant difference between the two groups. (2)TCM syndrome score and PADAM symptom score comparison:After treatment,both groups showed a significant reduction in TCM syndrome scores and PADAM symptom scores (P<0.01),and the observation group showed a significantly greater reduction compared to the control group (P<0.05). (3)Glucose metabolism indexes comparison:After treatment,the levels of FPG,2 h PG,and HbA1c were significantly reduced in both groups (P<0.01),and there was no statistically significant difference between the two groups regarding FPG,2 h PG,and HbA1c levels after treatment. (4)Serum sex hormone indexes comparison:After treatment,the levels of FT,TT,PRL,LH,and FSH were significantly increased in both groups (P<0.01),while E2 levels were significantly decreased (P<0.01). There was no statistically significant difference between the two groups in the levels of FT,TT,PRL,LH,FSH,and E2 after treatment. There was also no significant difference in SHBG levels within or between the groups before and after treatment. During the clinical observation,neither group exhibited any obvious adverse reactions. ConclusionModified Shibaotang can significantly improve the clinical symptoms of male late-onset hypogonadism of kidney essence deficiency syndrome complicated with diabetes mellitus,reduce blood glucose,and increase sex hormone levels. The mechanism may involve the inhibition of aromatase transformation in adipocytes,promotion of GnRH production,and regulation of the hypothalamic-pituitary-gonadal axis function.
6.Nutritional supply status of school meals for primary and secondary students in three cities of Henan Province
LI Shan, YANG Li, HE Qidong, CAO Linlin, CHEN Xiaolong, LIU Bingrui, FENG Yinhua, FU Pengyu
Chinese Journal of School Health 2025;46(1):50-52
Objective:
To assess the nutritional supply status of school meals for primary and secondary school students in Henan Province, so as to provide a basis for scientific guidance of school meals.
Methods:
During 2021-2023, 115 lunch and dinner samples were collected from 25 primary and secondary schools in Zhoukou, Anyang and Luoyang of Henan Province by a direct selection method, and 13 nutrients were determined for each sample. The nutrient supply was evaluated based on Nutrition Guidelines of School Meals and Reference Intake of Dietary Nutrients for Chinese Residents (2023 Edition). Mann-Whitney U test was used to compare the differences of nutritional supply between urban and rural schools.
Results:
The median values for energy (709.77 kcal,1 kcal=4.18 kJ), fat energy supply ratio (0.27) and carbohydrate energy supply ratio (0.55) in the 66 lunches and dinners from primary school were within the recommended range. The supply of protein (28.39 g) and sodium (1 464.59 mg) was excessive. The median values of zinc (2.62 mg) and dietary fiber (5.19 g) were lower than the reference values. No statistically significant differences were observed in the supply of 13 nutrients between urban and rural primary schools( U = 427.00 -633.00, P > 0.05 ). Among 49 samples from secondary schools, the median value of energy supply (930.02 kcal), carbohydrate energy ratio ( 0.54 ) and fat energy supply ratio(0.25) were within the recommended range; and the median values of protein (38.82 g) and sodium (2 556.80 mg) were higher than the standard; and the median values of calcium (250.32 mg) and vitamin B1 (0.16 mg) were lower than the standard. Additionally, the differences in the level of vitamin B2 ( U =372.00) and zinc ( U =375.00) between the urban and rural secondary schools were statistically significant ( P <0.05).
Conclusion
Nutrient supply of primary and secondary school meals in three cities of Henan Province is inadequate and imbalanced, and the recipe need to be further optimized and improved.
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.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.
9.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.
10.PAK4-PROTAC targeted degradation drug enhances immune cell-induced apoptosis in renal cell carcinoma
Chen YAO ; Bohan MA ; Xiaojing BAI ; Shan XU
Journal of Modern Urology 2025;30(6):527-532
Objective: To explore the potential application of PAK4-PROTAC targeted degradation drug (PpD) in renal cancer immunotherapy. Methods: TIMER 2.0 and TISIDB databases were used to analyze the relationship among PAK4 expression, tumor purity and abundance of immune cell infiltration in renal tumor microenvironment (TEM).Renal cancer cell lines OS-RC-2, 786-O and ACHN were treated with 0, 125 and 250 nmol/L PpD, and the effects of Jurkat cell co-culture on the results were investigated.The cell apoptosis was detected with flow cytometry, and the expression of programmed cell death 1 ligand 1 (PD-L1) in renal cancer cells was detected with immunoblotting. Results: The high expression of PAK4 was positively related to immune purity, and inhibited the abundance of immune killer cells in TEM, such as CD8 T cells, CD4 T cells, natural killer cells and dendritic cells.With 250 nmol/L PpD treatment, there were 21.02% apoptotic cells in OS-RC-2, 29.67% apoptotic cells in 786-O, and 15.39% apoptotic cells in ACHN, respectively.However, with the same concentration of 250 nmol/L PpD treatment, cell apoptotic rate was sharply increased to 70.13% in OS-RC-2/Jurkat, 70.68% in 780-O/Jurkat, and 60.27% in ACHN/Jurkat co-culture models, respectively. Conclusion: PpD can promote apoptosis of renal cancer cells by reducing the expression of PAK4 protein, and enhance the killing effects of immune cells on tumor cells.


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