OBJECTIVES: To investigate the mechanism by which Guijianyu ameliorates podocyte injury in a mouse model of diabetic kidney disease (DKD) induced by advanced glycation endproducts (AGEs). METHODS: Sixty db/db mouse models of DKD were randomized equally into 5 groups for treatment with saline, Guijianyu extract at 3 doses or irbesartan for 12 weeks, and the changes in renal pathology and structure were observed using transmission electron microscopy, and the expressions of related genes and key proteins were detected using RT-qPCR and immunohistochemistry. In cultured MPC-5 cells incubated with 50 mg/L AGEs-BSA for 24 h, the effect of different concentrations of Guijianyu extract on cell viability was examined with CCK-8 assay; Western blotting was performed to detect the protein expressions of RAGE, VEGFA, TNF-α, NF-κB(p65), IL-6 and caspase-3, and the mRNA expressions of RAGE, NF-κB (p65), VEGFA and IL-6 were detected with RT-qPCR. RESULTS: In mouse models of DKD, treatment with high-dose Guijianyu extract significantly reduced renal expressions of RAGE, VEGFA, NF-κB(p65), and IL-6 proteins and the mRNA expressions of RAGE, NF-κB, and IL-6. In MPC-5 cells, exposure to AGEs significantly reduced cell viability and increased the protein expressions of RAGE, NF‑κB (p65), VEGFA, TNF-α, IL-6 and caspase-3 (P<0.05) and mRNA expressions of RAGE, NF-κB (p65), VEGFA, and IL-6. Treatment with Guijianyu extract obviously improved cell viability and reduced the expressions of RAGE, NF-κB(p65), VEGFA, TNF-α, IL-6, and caspase-3. Furthermore, Guijianyu extract effectively reversed RAGE agonist-induced elevation of protein expressions of RAGE, VEGFA, TNF-α, IL-6, and caspase-3 and mRNA expressions of RAGE, NF-κB (p65), IL-6, and VEGFA in MPC-5 cells. CONCLUSIONS Guijianyu extract ameliorates AGEs-induced mouse renal podocyte injury in DKD by inhibiting the activation of AGEs-RAGE signaling pathway and reducing the expressions of pro-inflammatory cytokines and vascular endothelial growth factors.
Animals ; Glycation End Products, Advanced ; Drugs, Chinese Herbal/pharmacology* ; Mice ; Signal Transduction/drug effects* ; Podocytes/pathology* ; Diabetic Nephropathies/drug therapy* ; Receptor for Advanced Glycation End Products ; Vascular Endothelial Growth Factor A/metabolism* ; Interleukin-6/metabolism* ; Male

Objective:To investigate the indications for prenatal diagnosis using copy number variation-sequencing (CNV-seq) and the abnormalities detected by the method.Methods:This retrospective analysis involved 17 994 singleton pregnant women who underwent prenatal CNV-seq at the First Affiliated Hospital of Zhengzhou University from January 2019 to December 2022. These cases were divided into five groups based on the following indications for CNV-seq: abnormal fetal ultrasound findings, high-risk results indicated by non-invasive prenatal testing (NIPT) or Down's syndrome serological screening (Down's screening), adverse pregnancy history, and advanced maternal age. The proportions of cases with the indications for prenatal CNV-seq, the detection rates of abnormalities (numerical abnormalities of chromosomes, pathogenic/likely pathogenic CNV in structural abnormalities) in the five groups, and the distribution of these abnormalities were analyzed. Statistical analysis was performed using Chi-square test. Results:Among the 17 994 pregnant women, the women with abnormal fetal ultrasound findings, high-risk NIPT results, high-risk Down's screening results, adverse pregnancy history, and advanced maternal age accounted for 32.65% (5 875/17 994), 11.90% (2 142/17 994), 31.62% (5 690/17 994), 11.70% (2 105/17 994), and 12.13% (2 182/17 994), respectively. The detection rates of abnormalities in the five groups were 10.60% (623/5 875), 34.64% (742/2 142), 4.69% (267/5 690), 2.99% (63/2 105), and 3.67% (80/2 182), respectively. The overall detection rate of abnormalities was 9.86% (1 775/17 994). The cases with numerical abnormalities of chromosomes accounted for 68.79% (1 221/1 775), trisomy 21 was predominant (49.30%, 602/1 221). Chromosomal structural abnormalities were detected in 31.21% (554/1 775) of the cases with abnormalities, with 57.76% (320/554) harboring pathogenic CNVs and 42.24% (234/554) harboring likely pathogenic CNVs. The detection rate of chromosomal numerical abnormalities was higher than that of structural abnormalities in the abnormal fetal ultrasound group, NIPT high-risk group, and advanced maternal age group [6.81% (400/5 875) vs. 3.80% (223/5 875), χ2=53.10; 27.96% (599/2 142) vs. 6.68% (143/2 142), χ2=338.40; 2.43% (53/2 182) vs. 1.24% (27/2 182), χ2=8.61; all P<0.01]. A total of 416 microdeletions and 255 microduplications were detected in the 554 cases. The top three regions with the highest frequencies in microdeletions were Xp22.31 (12.74%, 53/416), 22q11.21 (7.93%, 33/416), and 17q12 (5.77%, 24/416); in microduplications, they were 22q11.21 (14.90%, 38/255), 17q12 (3.53%, 9/255), and 7q11.23 (3.53%, 9/255). Conclusions:Abnormal fetal ultrasound findings accounted for the highest proportion of prenatal diagnostic indications. The overall detection rate of abnormalities by CNV-seq is relatively high, especially in those with high-risk NIPT results as an indication for prenatal diagnosis. Among the chromosomal structural abnormalities detected in this study, the frequencies of Xp22.31 microdeletion and 22q11.21 microduplication are higher.

BACKGROUND:Erythropoietin has anti-inflammatory,anti-apoptotic,and pro-bone defect repair effects.To date,fewer studies have been conducted on its effects and molecular mechanism underlying restorative dentin formation after pulp injury. OBJECTIVE:To explore the effect of erythropoietin on restorative dentin formation after pulp injury. METHODS:(1)Animal experiment:Thirty-two rats were randomly divided into control group(n=16)and experimental group(n=16).In the experimental group,collagen sponges containing erythropoietin were used to directly cap the pulp at the pulp injury,and in the control group,collagen sponges containing PBS were used to directly cap the pulp at the exposed pulp injury.The cavity was then closed with glass ionomer adhesive.After 2 and 4 weeks of treatment,the maxillary bones of the two groups were collected,and the expression of nestin in dentin was detected by immunohistochemistry,and the reparative dentin production was observed by hematoxylin-eosin staining.The maxillae of four Sprague-Dawley rats were taken for immunohistochemical detection of erythropoietin expression in molar and incisor teeth.(2)Cell experiment:Human dental pulp cells,human periodontal ligament cells and human gingival fibroblasts were obtained from human dental tissue,periodontal ligament,and gingival tissue.Real-time reverse transcription PCR(RT-PCR)was used to detect the mRNA expression of erythropoietin.Erythropoietin,dentin sialophosphoprotein,dentin matrix protein 1,and nestin mRNA levels in human pulp cells were detected by RT-PCR under induced or uninduced odontoblastic differentiation.After down-regulation of erythropoietin expression or exogenous administration of erythropoietin intervention under induced or uninduced differentiation odontoblastic differentiation,the relative mRNA expression of dentin sialophosphoprotein and dentin matrix protein 1 in human pulp cells was detected by RT-PCR,and the formation of mineralized nodules was detected by alizarin red S staining,and mRNA and protein expressions of bone morphogenetic protein 2 were detected by RT-PCR and western blot,respectively. RESULTS AND CONCLUSION:(1)Animal experiment:Compared with the control group,the restorative dentin production and nestin expression were higher in the experimental group after 2 and 4 weeks of treatment.The expression of erythropoietin was weakly positive in pulp,odontoblastic cell layer and periodontal membrane of the rat's first maxillary molar,and strongly positive in odontoblasts.(2)Cell experiment:The mRNA expression of erythropoietin was higher in human dental pulp cells than in the other two types of cells.The mRNA expressions of dentin sialophosphorin,dentin matrix protein 1,nestin,erythropoietin and bone morphogenetic protein 2 in human pulp cells increased and the formation of mineralized nodules during odontoblastic differentiation under induction compared with non-induction conditions.The mRNA expression of dentin sialophosphoprotein,dentin matrix protein 1,nestin,bone morphogenetic protein 2 and the formation of mineralized nodules were decreased in human pulp cells after downregulation of erythropoietin under induced odontoblastic differentiation,and the protein expression of bone morphogenetic protein 2 was also decreased.After exogenous erythropoietin intervention,the expression of the above indexes in human dental pulp cells increased.To conclude,erythropoietin can promote the formation of dentin to some extent.

Objective To investigate the mechanism by which Guijianyu ameliorates podocyte injury in a mouse model of diabetic kidney disease(DKD)induced by advanced glycation endproducts(AGEs).Methods Sixty db/db mouse models of DKD were randomized equally into 5 groups for treatment with saline,Guijianyu extract at 3 doses or irbesartan for 12 weeks,and the changes in renal pathology and structure were observed using transmission electron microscopy,and the expressions of related genes and key proteins were detected using RT-qPCR and immunohistochemistry.In cultured MPC-5 cells incubated with 50 mg/L AGEs-BSA for 24 h,the effect of different concentrations of Guijianyu extract on cell viability was examined with CCK-8 assay;Western blotting was performed to detect the protein expressions of RAGE,VEGFA,TNF-α,NF-κB(p65),IL-6 and caspase-3,and the mRNA expressions of RAGE,NF-κB(p65),VEGFA and IL-6 were detected with RT-qPCR.Results In mouse models of DKD,treatment with high-dose Guijianyu extract significantly reduced renal expressions of RAGE,VEGFA,NF-κB(p65),and IL-6 proteins and the mRNA expressions of RAGE,NF-κB,and IL-6.In MPC-5 cells,exposure to AGEs significantly reduced cell viability and increased the protein expressions of RAGE,NF-κB(p65),VEGFA,TNF-α,IL-6 and caspase-3(P<0.05)and mRNA expressions of RAGE,NF-κB(p65),VEGFA,and IL-6.Treatment with Guijianyu extract obviously improved cell viability and reduced the expressions of RAGE,NF-κB(p65),VEGFA,TNF-α,IL-6,and caspase-3.Furthermore,Guijianyu extract effectively reversed RAGE agonist-induced elevation of protein expressions of RAGE,VEGFA,TNF-α,IL-6,and caspase-3 and mRNA expressions of RAGE,NF-κB(p65),IL-6,and VEGFA in MPC-5 cells.Conclusion Guijianyu extract ameliorates AGEs-induced mouse renal podocyte injury in DKD by inhibiting the activation of AGEs-RAGE signaling pathway and reducing the expressions of pro-inflammatory cytokines and vascular endothelial growth factors.

Objective:To evaluate the clinical efficacy of mitral valve transcatheter edge-to-edge repair (TEER) in patients with non-central degenerative mitral regurgitation (DMR).Methods:This retrospective study included patients with non-central DMR who underwent TEER at Fuwai Hospital between January 2021 and February 2024. Patients were categorized into two groups: the commissure-involved group and the non-commissure group, based on whether the mitral valve commissures were involved. Clinical data, surgical outcomes, and echocardiographic findings at 3 months postoperatively were collected and compared, and patients were followed up. The primary endpoint was the procedural success rate at discharge.Results:A total of 59 patients were included, aged (68.6±9.3) years, including 23 females (39%). In the overall study population, 78% (46/59) of patients had severe mitral regurgitation. Forty-two cases were in the non-commissure group, and 17 cases were in the commissure-involved group. Patients in the non-commissure group mainly had lesions in the A1/P1 region, while patients in the commissure-involved group mainly had lesions in the A3/P3 region. There was no significant difference in the procedural success rate at discharge (93% vs. 88%, P=0.95) and the incidence of postoperative complications (5% vs. 6%, P=1.00) between the two groups. Two patients in the commissure-involved group experienced single leaflet device attachment, with one of them requiring conversion to surgical mitral valve surgery; In the non-commissure group, two patients experienced single-valve clamping, and one of them was converted to surgical mitral valve surgery. The follow-up time of the entire cohort was (15.5±10.3) months. In the non-commissure group, 2 patients died and 2 were readmitted. While in the commissure-involved group, no patients died and only 1 patient was readmitted. Conclusion:TEER is an effective treatment for patients with non-central DMR involving the commissures, without increasing the incidence of postoperative complications.

Objective To study the pharmacokinetics of dihydralazine sulfate,triamterene,hydrochlorothiazide and reserpine in compound reserpine and triamterene in rats.Methods SD rats were randomly divided into three groups.Compound reserpine and triamterene was given at dose of 3.6,10.8 and 32.4 mg·kg-1 by single oral gavage,respectively.HPLC-MS was used to measure the blood concentrations of dihydralazine sulfate,triamterene,hydrochlorothiazide,and reserpine at various time points.DAS software was used to compute the pharmacokinetic parameters.Results After a single oral gavage of 3.6,10.8 and 32.4 mg·kg-1 of compound reserpine tablets triamterene,the tmax of dihydralazine sulfate in rat plasma were 1.50,1.33,and 1.42 h,and the Cmax of dihydralazine sulfate were 12.30,38.31 and 120.52 μg·L-1,respectively.The tmax of triamterene were 1.33,1.33,and 1.42 h,and the Cmax of triamterene were 20.93,67.36,and 168.64 μg·L-1,respectively.The tmax of hydrochlorothiazide were 2.00,2.00,and 1.75 h,and the Cmax of hydrochlorothiazide were 19.89,57.58,and 160.78 μg·L-1,respectively.Risperdal was found at very low levels in rat plasma,and only trace amounts were detected at 1.00 and 1.50 h of 32.4 mg·kg-1 administration.Conclusions Dihydralazine sulfate,triamterene,and hydrochlorothiazide can be eliminated quickly after compound reserpine and triamterene was given orally to rats,and their oral absorption is basically linear.The greater the dosage,the better the absorption of effective components.

Objective To explore the impact of integrated traditional Chinese and Western medicine on the overall survival(OS)of patients with biliary tract carcinoma(BTC),and to analyze the related factors affecting the OS of patients with BTC,so as to provide accurate prognosis assessment for clinical diagnosis and treatment.Methods The medical records of 198 patients with BTC in Henan Provincial Hospital of Traditional Chinese Medicine from February 2018 to October 2023 were retrospectively analyzed.The Cox proportional hazards model was utilized to analyze the independent prognostic factors affecting the OS of BTC patients.Survival curves were constructed using the Kaplan-Meier method,and the Log-rank test was employed to compare median OS differences among various groups.Results A total of 116 patients with BTC were included.There were 59 cases(50.86％)in the Western medicine group,with a median OS of(14±2.77)months,and the 1-,2-and 3-year OS rates were 81.36％,61.02％and 46.11％,respectively.There were 57 cases(49.14％)in the integrated traditional Chinese and Western medicine group,with a median OS of(29±3.91)months,and the 1-,2-and 3-year OS rates were 82.32％,67.64％and 58.00％,respectively.There was a statistically significant difference in survival time between the two groups(P=0.038).Univariate analysis indicated that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy and integrated traditional Chinese and Western medicine were significantly correlated with OS in BTC patients(all P＜0.05).Multivariate analysis revealed that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine had significant effects on the OS of patients with BTC(all P＜0.05).Conclusion The median OS of the traditional Chinese and Western medicine groups was significantly longer than that of the Western medicine group,and the long-term OS rate of the patients was significantly better than that of the Western medicine group.Age,clinical stage,surgery,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine are independent prognostic factors affecting OS in patients with BTC.

Objective:To investigate the indications for prenatal diagnosis using copy number variation-sequencing (CNV-seq) and the abnormalities detected by the method.Methods:This retrospective analysis involved 17 994 singleton pregnant women who underwent prenatal CNV-seq at the First Affiliated Hospital of Zhengzhou University from January 2019 to December 2022. These cases were divided into five groups based on the following indications for CNV-seq: abnormal fetal ultrasound findings, high-risk results indicated by non-invasive prenatal testing (NIPT) or Down's syndrome serological screening (Down's screening), adverse pregnancy history, and advanced maternal age. The proportions of cases with the indications for prenatal CNV-seq, the detection rates of abnormalities (numerical abnormalities of chromosomes, pathogenic/likely pathogenic CNV in structural abnormalities) in the five groups, and the distribution of these abnormalities were analyzed. Statistical analysis was performed using Chi-square test. Results:Among the 17 994 pregnant women, the women with abnormal fetal ultrasound findings, high-risk NIPT results, high-risk Down's screening results, adverse pregnancy history, and advanced maternal age accounted for 32.65% (5 875/17 994), 11.90% (2 142/17 994), 31.62% (5 690/17 994), 11.70% (2 105/17 994), and 12.13% (2 182/17 994), respectively. The detection rates of abnormalities in the five groups were 10.60% (623/5 875), 34.64% (742/2 142), 4.69% (267/5 690), 2.99% (63/2 105), and 3.67% (80/2 182), respectively. The overall detection rate of abnormalities was 9.86% (1 775/17 994). The cases with numerical abnormalities of chromosomes accounted for 68.79% (1 221/1 775), trisomy 21 was predominant (49.30%, 602/1 221). Chromosomal structural abnormalities were detected in 31.21% (554/1 775) of the cases with abnormalities, with 57.76% (320/554) harboring pathogenic CNVs and 42.24% (234/554) harboring likely pathogenic CNVs. The detection rate of chromosomal numerical abnormalities was higher than that of structural abnormalities in the abnormal fetal ultrasound group, NIPT high-risk group, and advanced maternal age group [6.81% (400/5 875) vs. 3.80% (223/5 875), χ2=53.10; 27.96% (599/2 142) vs. 6.68% (143/2 142), χ2=338.40; 2.43% (53/2 182) vs. 1.24% (27/2 182), χ2=8.61; all P<0.01]. A total of 416 microdeletions and 255 microduplications were detected in the 554 cases. The top three regions with the highest frequencies in microdeletions were Xp22.31 (12.74%, 53/416), 22q11.21 (7.93%, 33/416), and 17q12 (5.77%, 24/416); in microduplications, they were 22q11.21 (14.90%, 38/255), 17q12 (3.53%, 9/255), and 7q11.23 (3.53%, 9/255). Conclusions:Abnormal fetal ultrasound findings accounted for the highest proportion of prenatal diagnostic indications. The overall detection rate of abnormalities by CNV-seq is relatively high, especially in those with high-risk NIPT results as an indication for prenatal diagnosis. Among the chromosomal structural abnormalities detected in this study, the frequencies of Xp22.31 microdeletion and 22q11.21 microduplication are higher.

Objective To explore the impact of integrated traditional Chinese and Western medicine on the overall survival(OS)of patients with biliary tract carcinoma(BTC),and to analyze the related factors affecting the OS of patients with BTC,so as to provide accurate prognosis assessment for clinical diagnosis and treatment.Methods The medical records of 198 patients with BTC in Henan Provincial Hospital of Traditional Chinese Medicine from February 2018 to October 2023 were retrospectively analyzed.The Cox proportional hazards model was utilized to analyze the independent prognostic factors affecting the OS of BTC patients.Survival curves were constructed using the Kaplan-Meier method,and the Log-rank test was employed to compare median OS differences among various groups.Results A total of 116 patients with BTC were included.There were 59 cases(50.86％)in the Western medicine group,with a median OS of(14±2.77)months,and the 1-,2-and 3-year OS rates were 81.36％,61.02％and 46.11％,respectively.There were 57 cases(49.14％)in the integrated traditional Chinese and Western medicine group,with a median OS of(29±3.91)months,and the 1-,2-and 3-year OS rates were 82.32％,67.64％and 58.00％,respectively.There was a statistically significant difference in survival time between the two groups(P=0.038).Univariate analysis indicated that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy and integrated traditional Chinese and Western medicine were significantly correlated with OS in BTC patients(all P＜0.05).Multivariate analysis revealed that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine had significant effects on the OS of patients with BTC(all P＜0.05).Conclusion The median OS of the traditional Chinese and Western medicine groups was significantly longer than that of the Western medicine group,and the long-term OS rate of the patients was significantly better than that of the Western medicine group.Age,clinical stage,surgery,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine are independent prognostic factors affecting OS in patients with BTC.

Objective To study the pharmacokinetics of dihydralazine sulfate,triamterene,hydrochlorothiazide and reserpine in compound reserpine and triamterene in rats.Methods SD rats were randomly divided into three groups.Compound reserpine and triamterene was given at dose of 3.6,10.8 and 32.4 mg·kg-1 by single oral gavage,respectively.HPLC-MS was used to measure the blood concentrations of dihydralazine sulfate,triamterene,hydrochlorothiazide,and reserpine at various time points.DAS software was used to compute the pharmacokinetic parameters.Results After a single oral gavage of 3.6,10.8 and 32.4 mg·kg-1 of compound reserpine tablets triamterene,the tmax of dihydralazine sulfate in rat plasma were 1.50,1.33,and 1.42 h,and the Cmax of dihydralazine sulfate were 12.30,38.31 and 120.52 μg·L-1,respectively.The tmax of triamterene were 1.33,1.33,and 1.42 h,and the Cmax of triamterene were 20.93,67.36,and 168.64 μg·L-1,respectively.The tmax of hydrochlorothiazide were 2.00,2.00,and 1.75 h,and the Cmax of hydrochlorothiazide were 19.89,57.58,and 160.78 μg·L-1,respectively.Risperdal was found at very low levels in rat plasma,and only trace amounts were detected at 1.00 and 1.50 h of 32.4 mg·kg-1 administration.Conclusions Dihydralazine sulfate,triamterene,and hydrochlorothiazide can be eliminated quickly after compound reserpine and triamterene was given orally to rats,and their oral absorption is basically linear.The greater the dosage,the better the absorption of effective components.