Aim To explore the effect of the classical famous prescription Dahuang Zhechong pill(DHZCP)on relieving liver cirrhosis by regulating the stress fiber remodeling mediated by mechanistic signaling pathway and to explore the underlying mechanism.Methods Mice were randomly divided into the control group,model group,DHZCP low-dose group,DHZCP high-dose group,and Colchicine-positive control group.The liver cirrhosis mouse model was constructed by intrap-eritoneal injection of olive oil-solubilized CCl4.HE staining and serologic markers were used to reflect liver injury.Masson staining was used to evaluate collagen deposition in liver tissue.ELISA was applied to detect vasoactive molecules and cancer indicators.Atomic force microscopy was employed to detect liver tissue stiffness.Color Doppler diagnostic instrument was used to assess portal blood flow velocity.Western blot was utilized to detect ROCK2 expression and phosphoryla-tion of YAP,Cofilin,and MLC.Results The liver tis-sues in the model group had obvious inflammatory cell infiltration and collagen deposition,accompanied by significant elevation of serum transaminases and fibrosis indexes.Similarly,vasoactive molecules and cancer in-dicators were elevated,and the mechanoregulatory pro-tein ROCK2 expression and phosphorylation of Cofilin and MLC were elevated,with YAP being strongly de-phosphorylated.Both low and high doses of DHZCP re-versed the pathological changes,serological indices,and inhibited the activation of the stress fiber(SF)re-modeling mechanistic signaling pathway.Conclusion DHZCP effectively ameliorates liver tissue lesions in mice with liver cirrhosis,and its mechanism may be re-lated to the inhibition of SF remodeling mechanistic signaling pathway.

AIM To establish a GC-MS/MS method for the analysis of volatile components in Yinhu Ganmao Powder,and to determine the contents of α-pinene,camphene,sabinene,β-pinene,α-terpinene,(+)-limonene,p-cymene,1,8-cineole,linalool,L-menthol,terpinen-4-ol,DL-menthol,α-terpineol,tridecane,pulegone,caryophyllene,humulene,n-hexadecane and patchouli alcohol.METHODS The analysis was performed on a DB-624 UI capillary column(30 m×0.25 mm×1.40 μm ),and electron ionization source was adopted with multiple reaction monitoring mode.RESULTS Fifty volatile components and twenty-five liposoluble components were identified in volatile oils and medicinal material powder,respectively.Nineteen constituents showed good linear relationships within their own ranges(r ≥ 0.999 0),whose average recoveries were 84.43％-113.31％with the RSDs of less than 9.15％.CONCLUSION This stable,accurate and reproducible method can provide a reference for the quality evaluation of Yinhu Ganmao Powder.

Aim To explore the effect of the classical famous prescription Dahuang Zhechong pill(DHZCP)on relieving liver cirrhosis by regulating the stress fiber remodeling mediated by mechanistic signaling pathway and to explore the underlying mechanism.Methods Mice were randomly divided into the control group,model group,DHZCP low-dose group,DHZCP high-dose group,and Colchicine-positive control group.The liver cirrhosis mouse model was constructed by intrap-eritoneal injection of olive oil-solubilized CCl4.HE staining and serologic markers were used to reflect liver injury.Masson staining was used to evaluate collagen deposition in liver tissue.ELISA was applied to detect vasoactive molecules and cancer indicators.Atomic force microscopy was employed to detect liver tissue stiffness.Color Doppler diagnostic instrument was used to assess portal blood flow velocity.Western blot was utilized to detect ROCK2 expression and phosphoryla-tion of YAP,Cofilin,and MLC.Results The liver tis-sues in the model group had obvious inflammatory cell infiltration and collagen deposition,accompanied by significant elevation of serum transaminases and fibrosis indexes.Similarly,vasoactive molecules and cancer in-dicators were elevated,and the mechanoregulatory pro-tein ROCK2 expression and phosphorylation of Cofilin and MLC were elevated,with YAP being strongly de-phosphorylated.Both low and high doses of DHZCP re-versed the pathological changes,serological indices,and inhibited the activation of the stress fiber(SF)re-modeling mechanistic signaling pathway.Conclusion DHZCP effectively ameliorates liver tissue lesions in mice with liver cirrhosis,and its mechanism may be re-lated to the inhibition of SF remodeling mechanistic signaling pathway.

AIM To establish a GC-MS/MS method for the analysis of volatile components in Yinhu Ganmao Powder,and to determine the contents of α-pinene,camphene,sabinene,β-pinene,α-terpinene,(+)-limonene,p-cymene,1,8-cineole,linalool,L-menthol,terpinen-4-ol,DL-menthol,α-terpineol,tridecane,pulegone,caryophyllene,humulene,n-hexadecane and patchouli alcohol.METHODS The analysis was performed on a DB-624 UI capillary column(30 m×0.25 mm×1.40 μm ),and electron ionization source was adopted with multiple reaction monitoring mode.RESULTS Fifty volatile components and twenty-five liposoluble components were identified in volatile oils and medicinal material powder,respectively.Nineteen constituents showed good linear relationships within their own ranges(r ≥ 0.999 0),whose average recoveries were 84.43％-113.31％with the RSDs of less than 9.15％.CONCLUSION This stable,accurate and reproducible method can provide a reference for the quality evaluation of Yinhu Ganmao Powder.

Background and Objectives: Long-term pathological myocardial hypertrophy (MH) seriously affects the normal function of the heart. Dronedarone was reported to attenuate left ventricular hypertrophy of mice. However, the molecular regulatory mechanism of dronedarone in MH is unclear. Methods: Angiotensin II (Ang II) was used to induce cell hypertrophy of H9C2 cells.Transverse aortic constriction (TAC) surgery was performed to establish a rat model of MH.Cell size was evaluated using crystal violet staining and rhodamine phalloidin staining.Reverse transcription quantitative polymerase chain reaction and western blot were performed to detect the mRNA and protein expressions of genes. JASPAR and luciferase activity were conducted to predict and validate interaction between forkhead box O3 (FOXO3) and protein kinase inhibitor alpha (PKIA) promoter. Results: Ang II treatment induced cell hypertrophy and inhibited sirtuin 1 (SIRT1) expression, which were reversed by dronedarone. SIRT1 overexpression or PKIA overexpression enhanced dronedarone-mediated suppression of cell hypertrophy in Ang II-induced H9C2 cells. Mechanistically, SIRT1 elevated FOXO3 expression through SIRT1-mediated deacetylation of FOXO3 and FOXO3 upregulated PKIA expression through interacting with PKIA promoter. Moreover, SIRT1 silencing compromised dronedaronemediated suppression of cell hypertrophy, while PKIA upregulation abolished the influences of SIRT1 silencing. More importantly, dronedarone improved TAC surgery-induced MH and impairment of cardiac function of rats via affecting SIRT1/FOXO3/PKIA axis. Conclusions Dronedarone alleviated MH through mediating SIRT1/FOXO3/PKIA axis, which provide more evidences for dronedarone against MH.

Objective To investigate the regulatory effect and possible mechanism of lactic acid on the fibrotic phenotype of cardiac fibroblasts.Methods Mouse cardiac fibroblasts(mCFs)were divided into control group(conventional culture),experimental-L group(4 mmol·L-1 L-lactic acid),experimental-M group(8 mmol·L-1 L-lactic acid),experimental-H group(12 mmol·L-1 L-lactic acid),transforming growth factor-β1(TGF-β1)group(10 ng·mL-1 TGF-β1),combined group(10 ng·mL-1 TGF-β1+12 mmol·L-1 L-lactic acid)and monocarboxylate transporter inhibitor(CHC)group(3 mmol·L-1 CHC).Western blot was used to detect the expression of fibrosis-related proteins and pan-lactate modification(Pan Kla)and H3 histone K18 lactate modification;cell scratch assay was used to detect cell migration ability.Results The cell migration rates of the control group,TGF-β1 group,experimental-H group and combined group were(40.56±0.03)％,(61.61±0.04)％,(26.59±0.05)％and(38.33±0.06)％,respectively.Compared with the control group,TGF-β1 group and experimental-H group,TGF-β1 group and combined group,the differences were statistically significant(all P＜0.01).The relative expression levels of collagen type Ⅰ alpha 1(COL1A1)protein in the control group,TGF-β1 group,experimental-H group and TGF-β1+experimental-H group were 0.76±0.09,1.10±0.07,0.40±0.04 and 0.68±0.10,respectively;the relative expression levels of COL3A1 protein were 0.87±0.05,1.15±0.07,0.32±0.07 and 0.73±0.06,respectively;the relative expression levels of α-smooth muscle actin(α-SMA)protein were 0.86±0.04,1.24±0.09,0.30±0.05 and 0.74±0.08,respectively.Compared with the control group,the above indexes of the TGF-β1 group and the experimental-H group were significantly different from those of the control group,and the above indexes of the TGF-β1 group were significantly different from those of the combined group(all P＜0.01).The cell migration rates of mCFs in the control group,experimental-H group and CHC group were(62.60±6.50)％,(28.00±8.15)％and(39.40±4.50)％,respectively;the relative expression levels of COL1A1 protein were 1.10±0.07,0.49±0.04 and 0.34±0.06,respectively;the relative expression levels of COL3A1 protein were 1.04±0.10,0.60±0.20 and 0.37±0.03,respectively;the relative expression levels of α-SMA protein were 1.20±0.11,0.67±0.20 and 0.48±0.18,respectively;the modification levels of Pan Kla were 1.06±0.07,1.54±0.09 and 1.53±0.12,respectively;the modification levels of H3K18la protein were 0.67±0.06,1.23±0.06 and 1.14±0.08,respectively.The above indexes of CHC group and experimental-H group were significantly different from those of control group(all P＜0.01).Conclusion L-lactic acid may play a role in inhibiting the fibrosis phenotype of mCFs by increasing non-histone lactic acid modification and H3K18la modification.

Objective To investigate the regulatory effect and possible mechanism of lactic acid on the fibrotic phenotype of cardiac fibroblasts.Methods Mouse cardiac fibroblasts(mCFs)were divided into control group(conventional culture),experimental-L group(4 mmol·L-1 L-lactic acid),experimental-M group(8 mmol·L-1 L-lactic acid),experimental-H group(12 mmol·L-1 L-lactic acid),transforming growth factor-β1(TGF-β1)group(10 ng·mL-1 TGF-β1),combined group(10 ng·mL-1 TGF-β1+12 mmol·L-1 L-lactic acid)and monocarboxylate transporter inhibitor(CHC)group(3 mmol·L-1 CHC).Western blot was used to detect the expression of fibrosis-related proteins and pan-lactate modification(Pan Kla)and H3 histone K18 lactate modification;cell scratch assay was used to detect cell migration ability.Results The cell migration rates of the control group,TGF-β1 group,experimental-H group and combined group were(40.56±0.03)％,(61.61±0.04)％,(26.59±0.05)％and(38.33±0.06)％,respectively.Compared with the control group,TGF-β1 group and experimental-H group,TGF-β1 group and combined group,the differences were statistically significant(all P＜0.01).The relative expression levels of collagen type Ⅰ alpha 1(COL1A1)protein in the control group,TGF-β1 group,experimental-H group and TGF-β1+experimental-H group were 0.76±0.09,1.10±0.07,0.40±0.04 and 0.68±0.10,respectively;the relative expression levels of COL3A1 protein were 0.87±0.05,1.15±0.07,0.32±0.07 and 0.73±0.06,respectively;the relative expression levels of α-smooth muscle actin(α-SMA)protein were 0.86±0.04,1.24±0.09,0.30±0.05 and 0.74±0.08,respectively.Compared with the control group,the above indexes of the TGF-β1 group and the experimental-H group were significantly different from those of the control group,and the above indexes of the TGF-β1 group were significantly different from those of the combined group(all P＜0.01).The cell migration rates of mCFs in the control group,experimental-H group and CHC group were(62.60±6.50)％,(28.00±8.15)％and(39.40±4.50)％,respectively;the relative expression levels of COL1A1 protein were 1.10±0.07,0.49±0.04 and 0.34±0.06,respectively;the relative expression levels of COL3A1 protein were 1.04±0.10,0.60±0.20 and 0.37±0.03,respectively;the relative expression levels of α-SMA protein were 1.20±0.11,0.67±0.20 and 0.48±0.18,respectively;the modification levels of Pan Kla were 1.06±0.07,1.54±0.09 and 1.53±0.12,respectively;the modification levels of H3K18la protein were 0.67±0.06,1.23±0.06 and 1.14±0.08,respectively.The above indexes of CHC group and experimental-H group were significantly different from those of control group(all P＜0.01).Conclusion L-lactic acid may play a role in inhibiting the fibrosis phenotype of mCFs by increasing non-histone lactic acid modification and H3K18la modification.

OBJECTIVE To explore the efficacy of Sour Jujube Seed Decoction combined with Huanglian Wendan Decoction on adult chronic insomnia and its effect on hypnotic withdrawal.METHODS 187 patients with chronic insomnia were included for anal-ysis,including 102 in the traditional Chinese medicine(TCM)group and 85 in the western medicine group.The TCM group was trea-ted with Sour Jujube Seed Decoction combined with Huanglian Wendan Decoction,while the western medicine group was treated with benzodiazepine under the consideration of doctor.The intervention period was 1 month,with assessments using the Pittsburgh Sleep Quality Index(PSQI)conducted before and after the intervention.Follow-up evaluations were performed at 3 months and 6 months re-spectively after the intervention.RESULTS There was no significant difference between the two groups at baseline.After the inter-vention,the PSQI scores of patients in both groups were significantly improved(P<0.01).Among them,the TCM group was better than the western medicine group in the improvement of sleep quality and sleeping pills,total PSQI score reduction(P<0.01).The re-sults of linear regression analysis showed that after controlling for confounding factors,the regression coefficients of the TCM group in two different models were1.821 and 1.922 respectively,and the former was statistically significant(P<0.05).By screening patients who took hypnotics at baseline in the TCM group and comparing them with those in the western medicine group,the influencing factors of hypnotic withdrawal were analyzed.During the 3-month follow-up,25 out of 39 patients in the TCM group and 17 out of 80 patients in the western medicine group had hypnotic withdrawal(χ2= 19.25,P<0.001);during the 6-month follow-up,23 of the 39 patients in the TCM group and 18 of the 79 patients in the western medicine group had hypnotic withdrawal(χ2= 13.53,P<0.001),the with-drawal rate of patients in the TCM group was significantly higher than that in the western medicine group.Further regression analysis showed that after adjusting for confounding factors,the results showed that the western medicine group had a significantly higher rate of not withdrawal than the TCM group at 3 months(OR=5.50,95%CI:2.30～13.72)and 6 months(OR=6.43,95%CI:2.54～17.77),and the results were statistically different(P<0.05).CONCLUSION Sour Jujube Seed Decoction combined with Huangli-an Wendan Decoction is effective in treating adult chronic insomnia and assisting in hypnotic withdrawal.

This article analyzed the mechanism of Danggui Sini Decoction(DSD) in improving kidney injury caused by blood stasis syndrome(BSS) in rats. Firstly, 32 female SD rats were randomly divided into the following four groups: a normal group and a BSS group, both receiving an equal amount of distilled water by gavage; a normal+DSD group and a BSS+DSD group, both receiving 5.103 g·kg~(-1) DSD orally for a total of 14 days. Daily cold water bath was given to establish the BSS model, and on the 14th day, BSS rats were subcutaneously injected with 0.8 mg·kg~(-1) adrenaline. Normal rats were subjected to the water bath at 37 ℃ and injected with an equal volume of distilled water. After the experiment, 24-hour urine, serum, and kidney samples were collected for metabolomic analysis, biochemical measurements, and hematoxylin-eosin(HE) staining. The study then employed ~1H-NMR metabolomic technology to reveal the metabolic network regulated by DSD in improving BSS-induced kidney injury and used network pharmacology to preliminarily elucidate the key targets of the effectiveness of DSD. Pathological and biochemical analysis showed that DSD intervention significantly reduced inflammation and abnormal levels of blood creatinine, blood urea nitrogen, and urine protein in the kidneys. Metabolomic analysis indicated that DSD attenuated BSS-induced kidney injury primarily by regulating 10 differential metabolites and three major metabolic pathways(taurine and hypotaurine metabolism, citrate cycle, and acetaldehyde and dicarboxylic acid metabolism). Network pharmacology analysis suggested that the protective effect of DSD against BSS-induced kidney injury might be related to two key genes, ATP citrate lyase(ACLY) and nitric oxide synthase 2(NOS2), and two main metabolic pathways, i.e., arginine biosynthesis, and arginine and proline metabolism. This study, from the perspective of network regulation, provides initial insights and evidence into the mechanism of DSD in improving kidney injury induced by BSS, offering a basis for further investigation into the molecular mechanisms underlying its efficacy.
Rats ; Female ; Animals ; Rats, Sprague-Dawley ; Network Pharmacology ; Drugs, Chinese Herbal/chemistry* ; Metabolomics ; Kidney ; Arginine ; Water

OBJECTIVES: To improve the understanding of the clinical phenotypes and genetic characteristics of nephronophthisis (NPHP) and related syndromes in children. METHODS: A retrospective analysis was performed on the medical data of eight children with NPHP and related syndromes who were diagnosed and treated in the Department of Pediatrics of the Second Hospital of Hebei Medical University, from January 2018 to November 2022. The clinical characteristics and genetic testing results were analyzed. RESULTS: Among these eight children, there were five boys and three girls, with an age of onset ranging from 15 months to 12 years. All 8 children exhibited different degrees of renal function abnormalities when they attended the hospital. Among the eight children, two had the initial symptom of delayed development, two had the initial symptom of anemia, and two were found to have abnormal renal function during physical examination. The extrarenal manifestations included cardiovascular abnormalities in two children, skeletal dysplasia in two children, liver dysfunction in one child, retinitis pigmentosa in one child, and visceral translocation in one child. All eight children had renal structural changes on ultrasound, and four children had mild to moderate proteinuria based on routine urine test. Of all eight children, five had NPHP1 gene mutations and one each had a gene mutation in the NPHP3, IFT140, and TTC21B genes, and four new mutation sites were discovered. CONCLUSIONS Children with NPHP and related syndromes often have the initial symptom of delayed development or anemia, and some children also have extrarenal manifestations. NPHP and related syndromes should be considered for children with unexplained renal dysfunction, and high-throughput sequencing may help to make a confirmed diagnosis.
Child ; Humans ; Retrospective Studies ; Syndrome ; Kidney Diseases, Cystic/genetics* ; Mutation ; Phenotype