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 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 investigate the differences in clinical characteristics, diagnosis, and treatment of pediatric septic shock in pediatric intensive care unit (PICU) among hospitals of different levels. Methods: This retrospective study enrolled 368 children with septic shock treated in the PICU of Beijing Children's Hospital, Henan Children's Hospital, and Baoding Children's Hospital from January 2018 to December 2021. Their clinical data were collected, including the general information, location of onset (community or hospital-acquired), severity, pathogen positivity, consistence of guideline (the rate of standard attainment at 6 h after resuscitation and the rate of anti-infective drug administration within 1 h after diagnosis), treatment, and in-hospital mortality. The 3 hospitals were national, provincial, and municipal, respectively. Furthermore, the patients were divided into the tumor group and the non-tumor group, and into the in-hospital referral group and the outpatient or emergency admission group. Chi-square test and Mann-Whitney U test were used to analyze the data. Results: The 368 patients aged 32 (11, 98) months, of whom 223 were males and 145 females. There were 215, 107, and 46 patients with septic shock, with males of 141, 51, and 31 cases, from the national, provincial, and municipal hospitals, respectively. The difference in pediatric risk of mortality Ⅲ (PRISM Ⅲ) scores among the national,provincial and municipal group was statistically significant (26(19, 32) vs.19(12, 26) vs. 12(6, 19), Z=60.25,P<0.001). The difference in community acquired septic shock among the national,provincial and municipal group was statistically significant (31.6%(68/215) vs. 84.1%(90/107) vs. 91.3%(42/46), χ²=108.26,P<0.001). There were no significant differences in compliance with guidelines among the 3 groups (P>0.05). The main bacteria detected in the national group were Klebsiella pneumoniae (15.4% (12/78)) and Staphylococcus aureus (15.4% (12/78)); in the provincial group were Staphylococcus aureus (19.0% (12/63)) and Pseudomonas aeruginosa (12.7% (8/63)), and in the municipal group were Streptococcus pneumoniae (40.0% (10/25)) and Enteric bacilli (16.0% (4/25)). The difference in the proportion of virus and the proportion of 3 or more initial antimicrobials used among the national,provincial and municipal group was statistically significant (27.7% (43/155) vs. 14.9% (13/87) vs. 9.1% (3/33), 22.8%(49/215) vs. 11.2%(12/107) vs. 6.5%(3/46), χ²=8.82, 10.99, both P<0.05). There was no difference in the in-hospital mortality among the 3 groups (P>0.05). Regarding the subgroups of tumor and non-tumor, the national group had higher PRISM Ⅲ (31(24, 38) vs. 22 (21, 28) vs.16 (9, 22), 24 (18, 30) vs. 17(8, 24) vs. 10 (5, 16), Z=30.34, 10.45, both P<0.001), and it was the same for the subgroups of in-hospital referral and out-patient or emergency admission (29 (21, 39) vs. 23 (17, 30) vs. 15 (10, 29), 23 (17, 29) vs. 18 (10, 24) vs. 11 (5, 16), Z=20.33, 14.25, both P<0.001) as compared to the provincial and municipal group. There was no significant difference in the in-hospital mortality among the 2 pairs of subgroups (all P>0.05). Conclusion: There are differences in the severity, location of onset, pathogen composition, and initial antibiotics of pediatric septic shock in children's hospitals of different levels, but no differences in compliance with guidelines and in-hospital survival rate.
Female ; Male ; Humans ; Child ; Retrospective Studies ; Shock, Septic/therapy* ; Hospitalization ; Intensive Care Units, Pediatric ; Hospitals, Pediatric

Aim To prepare the sea cucumber enzy¬molysis fermentation liquid (SCEFL) by enzymatic hydrolysis of protease and fermentation of probiotics and to investigate the effect of SCEFL on the immunosup-pression induced by cyclophosphamide in mice and to explore its mechanism by metabomic method. Methods The immunosuppressive model was induced by in-traperitoneal injection of cyclophosphamide. C57BL/6J mice were randomly divided into normal group, model group, Levamisole group, SCEFL groups (at low, medium and high doses). The pathological changes of spleen were observed by HE staining. The proportion of CD4

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