1.Mechanism of Dangui Shaoyaosan in Alleviating Inflammatory Responses in Diabetic Kidney Disease by Modulating Macrophage Polarization in Kidneys of db/db Mice
Luyu HOU ; Linlin ZHENG ; Wenjing SHI ; Zixuan WANG ; Shilong GUO ; Zhe LYU ; Dengzhou GUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):1-10
ObjectiveTo observe the effects of Danggui Shaoyaosan on macrophage polarization and renal inflammation in db/db mice with diabetic kidney disease (DKD), and to explore its renal protective effects and underlying mechanisms. MethodsEight db/m mice were assigned to the normal group, and forty db/db mice were randomly divided into a model group, low-, medium-, and high-dose Danggui Shaoyaosan groups (8.39, 16.77, 33.54 g·kg-1), and an irbesartan group (0.025 g·kg-1). All mice were administered treatment by gavage for 12 consecutive weeks. General conditions of the mice were observed during the intervention. At the end of the 12-week intervention, 24-h urine samples were collected using metabolic cages, after which the mice were anesthetized for sample collection. Blood was collected by enucleation and centrifuged to obtain serum for the determination of glycated serum protein (GSP), serum creatinine (SCr), blood urea nitrogen (BUN), total cholesterol (TC), and triglycerides (TG). The urinary albumin-to-creatinine ratio (UACR) was measured. Renal pathological changes were observed using hematoxylin-eosin (HE) staining, periodic acid-Schiff (PAS) staining, and Masson staining. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and monocyte chemoattractant protein-1 (MCP-1) levels. Immunofluorescence (IF) was performed to detect F4/80 expression in renal tissue, and immunohistochemistry (IHC) was used to assess CD206 expression. Real-time quantitative polymerase chain reaction (Real-time PCR) was employed to measure the mRNA expression of TNF-α, IL-10, inducible nitric oxide synthase (iNOS), and arginase-1 (Arg-1). Western blot analysis was used to detect the protein expression of iNOS, Arg-1, CD86, and CD206 in renal tissue. ResultsCompared with the normal group, the model group showed increased levels of GSP, UACR, SCr, BUN, TC, and TG, elevated levels of the inflammatory factor TNF-α and the chemokine MCP-1, and decreased IL-10 levels (P<0.01). Pathological examination revealed glomerular hypertrophy, mesangial cell proliferation with marked mesangial expansion, inflammatory cell infiltration, vacuolar degeneration of renal tubular epithelial cells, prominent glycogen deposition, and increased collagen fiber deposition. In addition, relative F4/80 fluorescence intensity was enhanced, CD206 expression in the glomeruli and renal interstitium was reduced, and TNF-α and iNOS mRNA expression was increased. IL-10 and Arg-1 mRNA expression was decreased, iNOS and CD86 protein expression was increased, and Arg-1 and CD206 protein expression was decreased (P<0.05, P<0.01). Compared with the model group, the Danggui Shaoyaosan groups and the irbesartan group showed decreased levels of GSP, UACR, SCr, BUN, TC, and TG, reduced serum TNF-α and MCP-1 levels, and increased IL-10 levels. Renal pathological damage was improved to varying degrees. Relative F4/80 fluorescence intensity was reduced, CD206 expression in the glomeruli and renal interstitium was increased, and TNF-α and iNOS mRNA expression was decreased. IL-10 and Arg-1 mRNA expression was increased, iNOS and CD86 protein expression was reduced, and Arg-1 and CD206 protein expression was increased (P<0.05, P<0.01). ConclusionDanggui Shaoyaosan can improve renal function and alleviate renal pathological damage in db/db mice. Its mechanism may be related to inhibiting M1 pro-inflammatory macrophage polarization, promoting M2 anti-inflammatory macrophage polarization, reducing inflammatory responses, delaying the progression of renal fibrosis, improving renal pathological injury, and thereby exerting renal protective effects.
2.Clinical application value of the combined detection of fecal SDC2 and TFPI2 gene methylation in the diagnosis of colorectal cancer and advanced adenoma
Zhaorui WANG ; Xu CHEN ; Na LIU ; Qing WANG ; Kunfang SONG ; Linlin GUO ; Ping WANG ; Ningjing LEI
Chinese Journal of Clinical Laboratory Science 2025;43(10):732-736
Objective To investigate the clinical application value of the combined detection of fecal syndecan 2(SDC2)and tissue factor pathway inhibitor 2(TFPI2)gene methylation in the diagnosis of colorectal cancer and advanced adenoma.Methods 124 pa-tients with colorectal lesions,including 43 colorectal cancer,52 advanced adenoma,and 29 non-advanced adenoma,and 46 patients with other gastrointestinal diseases such as colorectal inflammatory diseases,ulcers,etc.,who visited 5 hospitals in Zhengzhou from 2020 to 2023 were enrolled as the disease group.In addition,78 healthy individuals who underwent physical examinations in the same period were selected as the healthy control group.Their fecal samples were collected before undergoing colonoscopy,and the methyla-tion levels of SDC2 and TFPI2 genes were detected using the quantitative methylation-specific PCR(qMSP).Results The positive rates of the combined detection of SDC2 and TFPI2 gene methylation in the colorectal cancer group and advanced adenoma group(95.34%and 88.46%)were significantly higher than those in the non-advanced adenoma group(48.27%,x2=18.75/13.63,P<0.05)and healthy controls(3.84%,x2=96.38/91.54,P<0.05).In the colorectal cancer group,the postoperative positive rate of the combined detection in 34 patients with positive methylation of SDC2 and TFPI2(5.88%)was significantly lower than the preoperative positive rate(95.34%,x2=54.74,P<0.05).Moreover,the mean Ct values of methylation of SDC2 and TFPI2 genes(32.38±3.72 and 31.85±3.21)in the colorectal cancer group were significantly lower than that in the advanced adenoma group(35.07±3.09 and 34.96±2.78,t=3.84 and 5.05,P<0.05).The analysis results of the ROC curve showed that the sensitivities of SDC2,TFPI2,and their combination detection in the diagnosis of colorectal cancer were 86.40%,88.37%,and 89.35%,respectively,while in differenti-ating advanced adenoma they were 73.52%,66.73%,and 80.15%,respectively.The combined detection of SDC2 and TFPI2 in differ-entiating colorectal cancer and advanced adenoma achieved the highest diagnostic efficacy(AUCROC=0.889 and 0.738,respectively).Conclusion The combined detection of methylation of SDC2 and TFPI2 genes in fecal samples has high diagnostic value for colorectal cancer and advanced adenomas,and also has certain clinical application value for the progression and postoperative monitoring of the disease.
3.Effects of SIRT2 on liver tight junctions and endoplasmic reticulum stress in cold-treated mice
Tianrui ZHAO ; Linlin XUE ; Junshu NIE ; Huaixiu ZHANG ; Chenchen LI ; Jingru GUO
Chinese Journal of Veterinary Science 2025;45(7):1469-1477
In order to investigate the effect of SIRT2 on tight junctions and endoplasmic reticulum stress in liver tissues of cold-treated mice,10 each of 5-week-old male C57BL/6 mice and SIRT2 knockout mice were selected and randomly divided into the wild-type room-temperature control group(WT Control),the wild-type cold-treated group(WT Cold),the SIRT2 knockout+room-temperature control group(KO Control)and SIRT2 knockout+cold treatment group(KO Cold).Mice in the room-temperature control group were kept at a temperature of(24+2)℃,and the cold-treatment group was placed in a(4+2)℃ artificial climate chamber for 3 h of random stimu-lation per day for 3 weeks.H&E staining,Masson staining,and transmission electron microscopy were employed to examine the microscopic and ultrastructural changes in mouse liver;AST and ALT concentrations in mouse serum were detected by biochemical analyzers;Western blot analysis was used to detect the expressions of tight junction-related proteins(Claudin1,Occludin),endo-plasmic reticulum stress-related proteins(GRP78,CHOP,XBP1,p-eIF2α,eIF2α),and pro-inflam-matory cytokines(TNF-α,IL-1β,IL-6).The results showed that compared with WT Control,the liver lobular structure of WT Cold and KO Control mice was unclear,hepatic cord arrangement was disordered,cytoplasm was loose,white vacuoles appeared,a small amount of collagen deposi-tion and fibroplasia,mitochondria were slightly swollen in hepatocytes,and endoplasmic reticulum was unevenly distributed,while the serum concentrations of AST and ALT were increased(P<0.050,P<0.010),and the liver tissues showed decreased protein expression of Occludin and Clau-din1(P<0.050,P<0.010,P<0.001),and increased protein expression of GRP78,CHOP,XBP1,p-eIF2α/eIF2α,TNF-α,IL-6,and IL-1β(P<0.050,P<0.010,P<0.001);compared with the KO Control,KO Cold mice showed a large number of white vacuoles,a small number of balloon-like lesions,inflammatory cell infiltration,obvious collagen deposition and fibroplasia,mitochondrial swelling in hepatocytes,mitochondrial ridge reduction,endoplasmic reticulum thickening,and ser-um AST and ALT concentrations increased(P<0.010),and in liver tissue,the protein expression of Occludin and Claudinl decreased(P<0.010),while the protein expression of GRP78,CHOP,XBP1,p-eIF2α/eIF2α,TNF-α,IL-6 and IL-1β increased(P<0.050,P<0.010,P<0.001).The re-sults showed that SIRT2 knockdown could aggravate the liver tissue tight junction damage caused by cold treatment,induce endoplasmic reticulum stress,and further promotes the inflammatory re-sponse.
4.Clinical application value of the combined detection of fecal SDC2 and TFPI2 gene methylation in the diagnosis of colorectal cancer and advanced adenoma
Zhaorui WANG ; Xu CHEN ; Na LIU ; Qing WANG ; Kunfang SONG ; Linlin GUO ; Ping WANG ; Ningjing LEI
Chinese Journal of Clinical Laboratory Science 2025;43(10):732-736
Objective To investigate the clinical application value of the combined detection of fecal syndecan 2(SDC2)and tissue factor pathway inhibitor 2(TFPI2)gene methylation in the diagnosis of colorectal cancer and advanced adenoma.Methods 124 pa-tients with colorectal lesions,including 43 colorectal cancer,52 advanced adenoma,and 29 non-advanced adenoma,and 46 patients with other gastrointestinal diseases such as colorectal inflammatory diseases,ulcers,etc.,who visited 5 hospitals in Zhengzhou from 2020 to 2023 were enrolled as the disease group.In addition,78 healthy individuals who underwent physical examinations in the same period were selected as the healthy control group.Their fecal samples were collected before undergoing colonoscopy,and the methyla-tion levels of SDC2 and TFPI2 genes were detected using the quantitative methylation-specific PCR(qMSP).Results The positive rates of the combined detection of SDC2 and TFPI2 gene methylation in the colorectal cancer group and advanced adenoma group(95.34%and 88.46%)were significantly higher than those in the non-advanced adenoma group(48.27%,x2=18.75/13.63,P<0.05)and healthy controls(3.84%,x2=96.38/91.54,P<0.05).In the colorectal cancer group,the postoperative positive rate of the combined detection in 34 patients with positive methylation of SDC2 and TFPI2(5.88%)was significantly lower than the preoperative positive rate(95.34%,x2=54.74,P<0.05).Moreover,the mean Ct values of methylation of SDC2 and TFPI2 genes(32.38±3.72 and 31.85±3.21)in the colorectal cancer group were significantly lower than that in the advanced adenoma group(35.07±3.09 and 34.96±2.78,t=3.84 and 5.05,P<0.05).The analysis results of the ROC curve showed that the sensitivities of SDC2,TFPI2,and their combination detection in the diagnosis of colorectal cancer were 86.40%,88.37%,and 89.35%,respectively,while in differenti-ating advanced adenoma they were 73.52%,66.73%,and 80.15%,respectively.The combined detection of SDC2 and TFPI2 in differ-entiating colorectal cancer and advanced adenoma achieved the highest diagnostic efficacy(AUCROC=0.889 and 0.738,respectively).Conclusion The combined detection of methylation of SDC2 and TFPI2 genes in fecal samples has high diagnostic value for colorectal cancer and advanced adenomas,and also has certain clinical application value for the progression and postoperative monitoring of the disease.
5.Effect of degree of neuromuscular block on intraoperative surgical conditions and postoperative recovery quality in patients undergoing lumbar interbody fusion
Jie GUO ; Xiaoyi CHEN ; Junda CHEN ; Xinghe WANG ; Dong HUANG ; Dongmei YUE ; Linlin ZHAO ; Junli CAO ; Su LIU
Chinese Journal of Anesthesiology 2025;45(4):444-448
Objective:To evaluate the effect of the degree of neuromuscular blockade on the intraoperative surgical conditions and postoperative recovery quality in patients undergoing lumbar interbody fusion.Methods:In this randomized controlled trial, 100 patients of either sex, aged 18-79 yr, with a body mass index of 18.5-35.0 kg/m 2, of American Society of Anesthesiologists Physical Status classification < Ⅳ, scheduled for elective lumbar interbody fusion at the Affiliated Hospital of Xuzhou Medical University from August to October 2024, were allocated into 2 groups ( n=50 each) using stratified randomization based on the number of lumbar segments: deep neuromuscular blockade group (group D) and moderate neuromuscular blockade group (group M). The intraoperative post-tetanic count was maintained at 1 or 2 in group D, and the intraoperative train-of-four was maintained at 1 or 2 in group M. The scores for surgical conditions, duration of operation, blood loss, length of incision, occurrence of severe hypoxemia after extubation, requirement for rescue analgesia in post-anesthesia care unit, 15-item Quality of Recovery scale score and length of stay were recorded. Results:Compared with group M, the scores for surgical conditions were significantly increased, the rate of rescue analgesia in post-anesthesia care unit was decreased, 15-item Quality of Recovery scale scores were increased at 3 days after surgery ( P<0.05), and no significant changes were found in the duration of operation, blood loss, length of incision, incidence of severe hypoxemia after extubation and length of hospital stay in group D ( P>0.05). Conclusions:Compared with moderate neuromuscular blockade, deep neuromuscular blockade can provide better surgical conditions and improve the quality of early postoperative recovery for patients undergoing lumbar interbody fusion.
6.Construction and validation of a risk prediction model for pneumothorax after CT-guided percutaneous lung puncture biopsy based on nomograms
Fanjie HAN ; Haibin WANG ; Linlin LI ; Ran GUO ; Changjiang LIU
The Journal of Practical Medicine 2025;41(15):2412-2417
Objective To construct and validate the efficacy of a risk prediction model for pneumothorax after CT-guided percutaneous pulmonary puncture biopsy(CT-PCNB)based on nomograms.Methods A total of 246 patients who underwent CT-PCNB examination in the hospital from October 2020 to October 2023 were selected and divided into training set(n=144)and validation set(n=102)using a random sampling method.In the training set,univariate and multivariate logistic regression analyses were performed to identify risk factors for pneumothorax after CT-PCNB.A nomogram model was constructed based on the identified risk factors,and its accuracy was validated using the validation set.Results Multifactorial logistic regression analysis showed that age≥60 years,concomitant underlying lung disease,lesion diameter<2 cm,distance from lesion to pleura≥10 mm,puncture through interlobular pleura,and≥2 pleural punctures were the risk factors for pneumothorax after CT-PCNB in the training set(P<0.05).A nomogram model was constructed based on these six factors.The ROC curve results for the training set showed an AUC of 0.852,sensitivity of 84.50%,and specificity of 67.50%.The nomogram model was validated using the validation set,with ROC curve results showing an AUC of 0.845,sensitivity of 83.00%,and specificity of 69.50%.There was no statistically significant difference between the predicted and actual values in both the training and validation sets(χ2=1.803,1.225;P>0.05),indicating clinical validity.Conclusion The nomogram model constructed based on the risk factors for pneumothorax after CT-PCNB has high predictive efficacy and is clinically meaningful.
7.Analysis of gene detection results of next-generation sequencing of liquid based cytological specimens of lung adenocarcinoma cavity effusion and evaluation of clinical efficacy
Shuo LIANG ; Yuan WANG ; Zihan SUN ; Jiameng ZHANG ; Xiaoyue XIAO ; Cong WANG ; Yue SUN ; Xinxiang CHANG ; Linlin ZHAO ; Huan ZHAO ; Huiqin GUO ; Zhihui ZHANG
Chinese Journal of Oncology 2025;47(9):905-912
Objective:To analyze the results of next generation sequencing (NGS) gene testing in liquid-based cytological specimens of lung adenocarcinoma cavity and evaluate the clinical efficacy of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) treatment.Methods:Liquid based cytological specimens of 222 cases of lung adenocarcinoma with cavity effusion and 201 cases of metastatic lymph node biopsy were collected. Specimens were obtained from the Cytology Laboratory of the Cancer Hospital of the Chinese Academy of Medical Sciences. The collection period was from January 2018 to December 2022. The results of NGS gene detection were compared. The clinical efficacy of 91 patients treated with EGFR-TKI was evaluated, and the survival curve was analyzed by Kaplan-Meier and other statistical methods.Results:The mutation rates of cancer-related genes detected by NGS were 82.0% (182/222) vs 79.1% (159/201), ( P=0.455) in liquid-based cytological specimens and histological specimens of metastatic lymph node biopsy, respectively. However, the mutation rate of EGFR T790M was significantly higher in cavity effusion than in lymph node biopsy specimens [12.2%(27/222)>3.5%(7/201), P=0.001]. The results of gene mutation were identical in 10 of the 13 cases with cavity effusion and metastatic lymph node biopsy, and the agreement rate of EGFR was 84.6%(11/13). In 3 inconsistent cases, EGFR mutations were detected in 2 cavity effusion cases that were not detected by lymph node biopsy. Results of genetic analysis of fluid-based cytological samples of 91 patients with cavity effusion were evaluated after drug treatment with EGFR-TKI. The mean progression-free survival (PFS) of the patients was 11.4 months (95% CI: 9.9-12.9). The mean PFS of patients harboring EGFR mutation was 12.3 months (95% CI: 10.8-13.9), and the mean PFS of EGFR wild type was 4.1 months (95% CI: 2.1-6.2). Conclusions:The results of NGS gene detection in liquid-based cytological specimens of lung adenocarcinoma patients with cavity effusion show that the PFS time is similar to that of histological specimens after clinical treatment with EGFR-TKI, which proves the reliability of NGS gene detection results in liquid cytological specimens. NGS gene testing appears higher sensitivity in cavity liquid-based samples than in metastatic lymph node samples.
8.Construction and validation of a risk prediction model for pneumothorax after CT-guided percutaneous lung puncture biopsy based on nomograms
Fanjie HAN ; Haibin WANG ; Linlin LI ; Ran GUO ; Changjiang LIU
The Journal of Practical Medicine 2025;41(15):2412-2417
Objective To construct and validate the efficacy of a risk prediction model for pneumothorax after CT-guided percutaneous pulmonary puncture biopsy(CT-PCNB)based on nomograms.Methods A total of 246 patients who underwent CT-PCNB examination in the hospital from October 2020 to October 2023 were selected and divided into training set(n=144)and validation set(n=102)using a random sampling method.In the training set,univariate and multivariate logistic regression analyses were performed to identify risk factors for pneumothorax after CT-PCNB.A nomogram model was constructed based on the identified risk factors,and its accuracy was validated using the validation set.Results Multifactorial logistic regression analysis showed that age≥60 years,concomitant underlying lung disease,lesion diameter<2 cm,distance from lesion to pleura≥10 mm,puncture through interlobular pleura,and≥2 pleural punctures were the risk factors for pneumothorax after CT-PCNB in the training set(P<0.05).A nomogram model was constructed based on these six factors.The ROC curve results for the training set showed an AUC of 0.852,sensitivity of 84.50%,and specificity of 67.50%.The nomogram model was validated using the validation set,with ROC curve results showing an AUC of 0.845,sensitivity of 83.00%,and specificity of 69.50%.There was no statistically significant difference between the predicted and actual values in both the training and validation sets(χ2=1.803,1.225;P>0.05),indicating clinical validity.Conclusion The nomogram model constructed based on the risk factors for pneumothorax after CT-PCNB has high predictive efficacy and is clinically meaningful.
9.Effect of Serum Containing Buyang Huanwutang on Podocytes Induced by High Glucose in Mice
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):64-73
ObjectiveTo observe the effect of serum containing Buyang Huanwutang on podocyte injury induced by high glucose in mice, and to explore its potential mechanism. MethodsMouse podocytes (MPC5) were cultured in vitro, and the optimal intervention concentration and time of serum containing Buyang Huanwutang were screened. Cells were divided into normal group (5.5 mmol·L-1 glucose), isotonic group (5.5 mmol·L-1 glucose+24.5 mmol·L-1 Mannitol), high glucose group (30 mmol·L-1 glucose), blank serum group (30 mmol·L-1 glucose+20% blank serum) and Buyang Huanwutang(30 mmol·L-1 glucose+10% serum containing). FerroOrange fluorescent probe was used to detect the level of iron (Fe2+) in cells. The level of reactive oxygen species (ROS) in cells was detected by Hoechst staining. The levels of glutathione (GSH) and malondialdehyde (MDA) in cells were detected by the kit. Enzyme-linked immunosorbent assay (ELISA) was used to determine the level of 4-hydroxynonenal (4-HNE). Western blot was used to detect Desmin, podocyte hole membrane protein (Nephrin), Podocalyxin, long-chain acyl-CoA synthetase 4(ACSL4), member 11 of solute carrier family 7 (SLC7A11), glutathione peroxidase 4(GPX4), nuclear transcription factor E2-related factor 2(Nrf2) and heme oxygenase -1(HO-1). The mRNA expression levels of Desmin, Nephrin, Podocalyxin, ACSL4, SLC7A11, GPX4, Nrf2 and HO-1 were detected by real-time fluorescence quantitative polymerase chain reaction. ResultsCompared with the normal control group, the expressions of Desmin and ACSL4, the levels of Fe2+ and ROS, and the contents of MDA and 4-HNE in the high glucose group increased significantly, and the expressions of GSH, Nephrin, Podocalyxin, SLC7A11, GPX4, Nrf2 and HO-1 decreased(P<0.05, P<0.01). There was no significant change in the above results in isotonic group. Compared with the high glucose group, the expressions of Desmin and ACSL4, the levels of Fe2+ and ROS, and the contents of MDA and 4-HNE in Buyang Huanwutang-containing serum group and Fer-1 group decreased significantly, and the expressions of GSH, Nephrin, Podocalyxin, SLC7A11, GPX4, Nrf2 and HO-1 increased(P<0.05, P<0.01). There was no significant change in the above results in blank serum group. ConclusionBuyang Huanwutang medicated serum can alleviate podocyte injury induced by high glucose, and its mechanism is related to regulating Nrf2/HO-1 pathway and inhibiting ferroptosis.
10.Treatment of Recurrent Pediatric Cough and Asthma Based on the Theory of "Mutual Dependence of Ascending and Descending" from the Perspectives of Deficiency,Phlegm,and Blood Stasis
Yu LIU ; Ying DING ; Yongbin YAN ; Xuran GUO ; Linlin LIU
Journal of Traditional Chinese Medicine 2025;66(9):963-967
Guided by the theory of "mutual dependence of ascending and descending", recurrent pediatric cough and asthma are considered to result from the imbalance of the ascending and descending of zang-fu organ qi, which is closely associated with deficiency, phlegm, and blood stasis. In clinical practice, the disease is treated based on differentiation during the initial stage, progression stage, and stable stage. In the initial stage, the pathogenesis is attributed to lung deficiency complicated by external pathogens and liver qi rising to attack the lung; the treatment should focus on restraining the lung and calming the liver, using the self-formulated Longchai Yuchuan Fomulation (龙柴愈喘方). During the progression stage, the disorder is due to the spleen failing to ascend clear qi and the kidney failing to grasp qi, leading to phlegm formation from deficiency; treatment should focus on tonifying spleen yang, supplementing kidney qi, and resolving phlegm and fluid retention, using a modified combination of Linggui Zhugan Decoction (苓桂术甘汤) and Jinkui Shenqi Pill (金匮肾气丸). In the stable stage, qi deficiency leads to poor consolidation, with phlegm turbidity and blood stasis; the treatment should aim at tonifying qi, transforming phlegm, and dispelling blood stasis, using a modified version of Guizhi Fuling Pill (桂枝茯苓丸).

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