Emodin is a hydroxyanthraquinone compound that is widely distributed and has multiple pharmacological activities, including anti-diarrheal, anti-inflammatory, and liver-protective effects. Research indicates that emodin may be one of the main components responsible for inducing hepatotoxicity. However, studies on the mechanisms of liver injury are relatively limited, particularly those related to bile acids(BAs) metabolism. This study aims to systematically investigate the effects of different dosages of emodin on BAs metabolism, providing a basis for the safe clinical use of traditional Chinese medicine(TCM)containing emodin. First, this study evaluated the safety of repeated administration of different dosages of emodin over a 5-week period, with a particular focus on its impact on the liver. Next, the composition and content of BAs in serum and liver were analyzed. Subsequently, qRT-PCR was used to detect the mRNA expression of nuclear receptors and transporters related to BAs metabolism. The results showed that 1 g·kg~(-1) emodin induced hepatic damage, with bile duct hyperplasia as the primary pathological manifestation. It significantly increased the levels of various BAs in the serum and primary BAs(including taurine-conjugated and free BAs) in the liver. Additionally, it downregulated the mRNA expression of farnesoid X receptor(FXR), retinoid X receptor(RXR), and sodium taurocholate cotransporting polypeptide(NTCP), and upregulated the mRNA expression of cholesterol 7α-hydroxylase(CYP7A1) in the liver. Although 0.01 g·kg~(-1) and 0.03 g·kg~(-1) emodin did not induce obvious liver injury, they significantly increased the level of taurine-conjugated BAs in the liver, suggesting a potential interference with BAs homeostasis. In conclusion, 1 g·kg~(-1) emodin may promote the production of primary BAs in the liver by affecting the FXR-RXR-CYP7A1 pathway, inhibit NTCP expression, and reduce BA reabsorption in the liver, resulting in BA accumulation in the peripheral blood. This disruption of BA homeostasis leads to liver injury. Even doses of emodin close to the clinical dose can also have a certain effect on the homeostasis of BAs. Therefore, when using traditional Chinese medicine or formulas containing emodin in clinical practice, it is necessary to regularly monitor liver function indicators and closely monitor the risk of drug-induced liver injury.
Emodin/administration & dosage* ; Bile Acids and Salts/metabolism* ; Animals ; Male ; Liver/injuries* ; Chemical and Drug Induced Liver Injury/genetics* ; Drugs, Chinese Herbal/adverse effects* ; Humans ; Rats, Sprague-Dawley ; Mice ; Rats

OBJECTIVES: To investigate the molecular epidemiology of children with phenylketonuria (PKU) in Gansu, China, providing foundational data for intervention strategies. METHODS: A retrospective analysis was conducted on 1 159 PKU families who attended Gansu Provincial Maternity and Child Care Hospital from January 2012 to December 2024. Sanger sequencing, multiplex ligation-dependent probe amplification, whole exome sequencing, and deep intronic variant analysis were used to analyze the PAH gene. RESULTS: For the 1 159 children with PKU, 2 295 variants were identified in 2 318 alleles, resulting in a detection rate of 99.01%. The detection rates were 100% (914/914) in 457 classic PKU families, 99.45% (907/912) in 456 mild PKU families, and 96.34% (474/492) in 246 mild hyperphenylalaninemia families. The 2 295 variants detected comprised 208 distinct mutation types, among which c.728G>A (14.95%, 343/2 295) had the highest frequency, followed by c.611A>G (4.88%, 112/2 295) and c.721C>T (4.79%, 110/2 295). The cumulative frequency of the top 23 hotspot variants reached 70.28% (1 613/2 295), and most variant alleles were detected in exon 7 (29.19%, 670/2 295). CONCLUSIONS Deep intronic variant analysis of the PAH gene can improve the genetic diagnostic rate of PKU. The development of targeted detection kits for PAH hotspot variants may enable precision screening programs and enhance preventive strategies for PKU.
Humans ; Phenylketonurias/epidemiology* ; Female ; Male ; Retrospective Studies ; Phenylalanine Hydroxylase/genetics* ; Mutation ; Child, Preschool ; China/epidemiology* ; Child ; Infant

This study established a droplet digital PCR(ddPCR)EvaGreen assay and probe methods for Schistosoma japonicum detection,and evaluated their application in detecting early infections in the S.japonicum host oncomelania and mice.Primers and corresponding probes for both ddPCR methods were designed and synthesized,and plasmids containing target sequences were constructed.The sensitivity of the two methods was tested through detection of the corresponding plasmids,and infectious and mixed oncomelania genomic DNA.Their specificity was evaluated by the detection of genomic DNA of negative oncomelania,Schistosoma mansoni,Clonorchis sinensis,Spirometra mansoni,and S.japonicum(as a positive control).The ddPCR probe method was evaluated by detection of early infection of oncomelania exposed tomiracidium with various ratios and incubation times,and the early migration and distribution of cercaria or schistosomula in mouse hosts infected with 200 cercaria via abdominal skin contact.According to standard curves constructed through the detection of plasmid serial dilutions,the regression equation for the EvaGreen assay was y=-0.839 9x+7.050 9,with a correlation coefficient R2=0.988 1,and the regression equation for the probe method was y=-1.047 5x+7.255 1,with a correlation coefficient R2=0.999 8.The lowest limit of plasmid detection with the probe method was between 38.94 cp/μL and 194.74 cp/μL.Both methods successfully detected positive reactions in the genomic DNA samples of infectious oncomelania at concentrations above 0.002 ng/μL and in the genomic DNA of each group of oncomelania mixtures.No significant differences between probe methods were observed in the detection values in the control group and the genomic DNA of negative oncomelania,S.mansoni,C.sinensis,and S.mansoni.However,the detection value of genomic DNA of negative oncomelania(291 ng/μL)with the EvaGreen assay was(20.3±4.39)cp/μL,a value significantly higher than the(1.5±0.1)cp/μL observed in the control group.For detection of early infection in oncomelania,the probe method detected Schistosoma japonicum DNA after 30 s incubation at room temperature with a≥5∶1 ratio of miracidium to oncomelania;the detection value peaked after a short time(5 min),and the peak value showed a fold increase similar to the increase in the miracidium to oncomelania ratio.Detection of early stage infection in mice with the probe method revealed that the schistosomula entered the lungs on day 2 and the liver on day 4,and continually migrated within the organs with abundant blood supply(spleen,kidney,and brain)in the first 9 days;moreover,a tendency toward ectopic parasitism was observed in the heart and pancreas on day 9,and a constantly negative control level was observed in the testes.The ddPCR probe method was more accurate and specific than the EvaGreen assay in the detection of plasmids,and infectious and mixed oncomelania,and the latter showed non-specific reactions in negative oncomelania detection.In a practical application,the probe method was demonstrated to be sensitive,to effectively reflect the early infection of oncomelania,and to reveal schistosomula migration and distribution in multiple organs of infected mice.

Objective By observing and measuring the relevant data of the buccal branch of the facial nerve,the parotid duct and the facial artery,the positional relationship among the three was analyzed to avoid accidental injury to the buccal branch of the facial nerve and the parotid duct when ligaturing the facial artery during the operation.Methods Forty adult head and neck specimens were dissected to observe the relationship between the buccal branch of the facial nerve and the parotid duct,the course and positional relationship of the facial artery,and the relationship between the buccal branch of the facial nerve and the peripheral vascular network.The relevant diameters were measured with a vernier caliper.Results The buccal branch of the facial nerve was divided into the superior buccal branch and the inferior buccal branch,and there was no direct anastomosis or connecting fiber between the buccal branch of the facial nerve and the parotid duct.The superior buccal branch was relatively thick,and it has a relatively constant position,which was parallel to the parotid duct.The position of the inferior buccal branch was not constant and it ran on or slightly above the plane of angulus oris.The superior buccal branch was located(10.76±5.54)mm from the parotid duct,while the inferior buccal branch was positioned(6.84±4.06)mm away from the parotid duct.The course of the main trunk of the facial artery was relatively fixed.Moreover,if the branch of the facial artery was missing,other branches of the facial artery would extend to replace the missing branch artery.The main trunk of the facial artery had a diameter of(2.34±0.83)mm,and its branches formed anastomoses with the buccal branch of the maxillary artery,creating a vascular network in the parotid and buccal regions.There was a vascular network around the buccal branch of the facial nerve,which was mostly small branches of the facial artery and the superficial temporal artery.Conclusion The buccal branch of the facial nerve exhibits a consistent anatomic relationship with the parotid duct and the facial artery.During the ligation of the facial artery,the parotid duct can serve as a landmark to accurately locate the buccal branch of the facial nerve,thereby significantly reducing the risk of inadvertent injury to the buccal branch of the facial nerve and the parotid duct.

Objective To explore the relationship between the level of Caveolin-1 and high mobility group protein B1(HMGB-1)in cerebrospinal fluid and the severity of sepsis and their predictive value.Methods A total of 102 children with sepsis were selected in a hospital from June 2021 to December 2023.According to neonatal critical case scoring criteria,the children were divided into mild group(n=41)and severe group(n=61).According to the diagnostic criteria of purulent meningitis and the results of cerebrospinal fluid examina-tion,the children were divided into sepsis complicated with purulent meningitis group(n=16)and simple sepsis group(n=86).The clinical data of the children were collected,and the levels of Caveolin-1,HMGB-1 and inflammatory factors[hypersensitive C-reactive protein(hs-CRP),procalcitonin(PCT)and tumor necro-sis factor-α(TNF-α)]in cerebrospinal fluid of the children were detected by enzyme-linked immunosorbent assay.Multivariate Logistic regression analysis was conducted to analyze the factors affecting the severity of sepsis,and the receiver operating characteristic curve was drawn to analyze the diagnostic value of Caveolin-1 and HMGB-1 in the severity of sepsis and the prediction value of purulent meningitis in the children.Results Compared with mild group,cerebrospinal fluid levels of Caveolin-1 and HMGB-1 in severe group were signifi-cantly increased,and the difference was statistically significant(P<0.05).The levels of PCT,hs-CRP and TNF-α in severe group were significantly higher than those in mild group,and the difference was statistically significant(P<0.05).Multivariate Logistic regression analysis showed that PCT,hs-CRP,TNF-α,Caveolin-1 and HMGB-1 levels were all risk factors for the severity of sepsis in children(P<0.05).The combined diag-nosis of Caveolin-1 and HMGB-1 was significantly better than that of Caveolin-1(Z=2.109,P=0.035),HMGB-1(Z=2.099,P=0.036),PCT(Z=2.487,P=0.013),hs-CRP(Z=2.419,P=0.016)and TNF-α(Z=3.441,P=0.001)were diagnosed alone.Compared with simple sepsis group,cerebrospinal fluid Caveo-lin-1 and HMGB-1 levels in sepsis complicated with purulent meningitis group were significantly increased,and the difference was statistically significant(P<0.05).The combined prediction of Caveolin-1 and HMGB-1 was significantly better than that of Caveolin-1(Z=2.621,P=0.009)and HMGB-1(Z=1.997,P=0.046)alone.Conclusion There are significant increases in the levels of Caveolin-1 and HMGB-1 in chil-dren with severe sepsis or sepsis complicated with purulent meningitis.Caveolin-1 and HMGB-1 have a certain clinical value in evaluating the severity of sepsis in children and predicting whether they are complicated with purulent meningitis.

Background: Aristolochic acid II (AAII), a major nephrotoxic and carcinogenic component of aristolochic acids (AAs), has been less studied compared with its well-characterized analog, aristolochic acid I (AAI). Although AAs are known to induce carcinogenesis via DNA adduct formation, the toxicity mechanisms, environmental prevalence, and long-term health impacts of AAII remain poorly understood. Objective: This study aimed to systematically evaluate AAII’s acute and chronic toxicity, carcinogenic mechanisms, and environmental exposure patterns using integrated murine models and phytochemical analyses to clarify its toxicological profile and associated health risks. Methods: C57BL/6J mice were used in the following experiments: (1) determination of AAII content in 3 commonly used Aristolochia medicinal materials via liquid chromatography-mass spectrometry/mass spectrometry; (2) acute toxicity testing with single doses of 10, 20, or 40 mg/kg; and (3) chronic exposure with 1 or 10 mg/kg administered every other day for 24 weeks, followed by 21 to 40 weeks of postexposure monitoring. Histopathological examination, whole-exome sequencing, biochemical assays, and micronucleus tests were performed to assess multi-organ damage, tumorigenesis, genomic mutation signatures, and direct clastogenicity. Phytochemical analyses were used to evaluate environmental distribution. Results: (1) A single 40 mg/kg dose of AAII induced dose-dependent renal tubular degeneration without hepatotoxicity; (2) the 10 mg/kg group showed significant mortality (20%), tumor incidence (33.3%, primarily forestomach and bladder transitional cell carcinomas), persistent renal interstitial fibrosis, and subclinical hepatic injury. Chronic exposure to 1 mg/kg still induced 13.3% mortality and 15.5% tumor incidence over a 64-week period; (3) whole-exome sequencing revealed a predominance of C>T mutations and pathway enrichment in chemical carcinogenesis and cytochrome P450-mediated metabolism, indicating reactive metabolite-driven mechanisms distinct from classical AA-DNA adducts; and (4) no histopathological changes were observed in nontarget organs (brain, heart, and testes), and micronucleus assays confirmed the absence of direct clastogenicity. Conclusion: This study highlights the delayed carcinogenic risks of low-dose chronic AAII exposure and emphasizes the need to update regulatory frameworks to ensure the safe use of aristolochiaceae-containing herbal products.

Objective To investigate the expression of serum microRNA(miR)-16a and mir-15b in patients with early-onset neonatal sepsis(EONS)and its value in clinical diagnosis and prognosis evaluation.Methods 114 children diagnosed with EONS admitted to the Department of Neonatology,Baoding Hospital of Beijing Children's Hospital Affiliated to Capital Medical University from October 2020 to October 2023 were selected as the EONS group,and 114 healthy newborns in the hospital during the same period were selected as the control group.Follow up was conducted on all EONS patients for a period of 6 months,including 34 cases in the poor prognosis group and 80 cases in the good prognosis group.The relative expression levels of miR-16a and miR-15b were measured within 6h of admission using real-time fluorescence quamtitative PCR(qRT-PCR).ROC curve was applied to analyze the diagnostic and prognostic value of serum miR-16a and miR-15b for EONS.Multivariate Logistic regression was applied to analyze the factors that affected the poor prognosis of EONS.Results Compared with the reference group,the birth weight of children in the EONS group(3.44±0.35kg/m2 vs 3.89±0.40kg/m2)was obviously reduced,and there was a obvious difference(35/114,1/114)in pathological jaundice between the two groups,and the differences were statistically significant(t=9.040,χ2=35.922,all P＜0.05).Compared with the reference group,the serum levels of miR-16a and miR-15b in the EONS group were obviously increased,and the differences was statistically significant(t=7.699,8.606,all P＜0.05).The area under the curve(AUC)of the ROC curve for the combined diagnosis of EONS by serum miR-16a and miR-15b was higher than that of miR-16a and miR-15b alone,and the differences were statistically significant(Z=2.619,2.157,all P<0.05).Compared with the good prognosis group,the poor prognosis group had a obvious decrease in body weight at birth,and a obvious increase in serum levels of miR-16a and miR-15b,and the differences were statistically significant(t=5.434,6.308,all P＜0.05).The AUC of serum miR-16a and miR-15b combined for prognostic evaluation was higher than that of miR-16a and miR-15b evaluated separately,and the differences were statistically significant(Z=2.364,2.073,all P＜0.05).Birth weight,miR-16a,miR-15b expression level was a prognostic factor of EONS(Wald χ2=33.459,16.146,49.797,all P<0.05).Conclusion MiR-16a and miR-15b are highly expressed in the serum of children with EONS,and they have certain diagnostic and prognostic value for EONS.

Objective By observing and measuring the relevant data of the buccal branch of the facial nerve,the parotid duct and the facial artery,the positional relationship among the three was analyzed to avoid accidental injury to the buccal branch of the facial nerve and the parotid duct when ligaturing the facial artery during the operation.Methods Forty adult head and neck specimens were dissected to observe the relationship between the buccal branch of the facial nerve and the parotid duct,the course and positional relationship of the facial artery,and the relationship between the buccal branch of the facial nerve and the peripheral vascular network.The relevant diameters were measured with a vernier caliper.Results The buccal branch of the facial nerve was divided into the superior buccal branch and the inferior buccal branch,and there was no direct anastomosis or connecting fiber between the buccal branch of the facial nerve and the parotid duct.The superior buccal branch was relatively thick,and it has a relatively constant position,which was parallel to the parotid duct.The position of the inferior buccal branch was not constant and it ran on or slightly above the plane of angulus oris.The superior buccal branch was located(10.76±5.54)mm from the parotid duct,while the inferior buccal branch was positioned(6.84±4.06)mm away from the parotid duct.The course of the main trunk of the facial artery was relatively fixed.Moreover,if the branch of the facial artery was missing,other branches of the facial artery would extend to replace the missing branch artery.The main trunk of the facial artery had a diameter of(2.34±0.83)mm,and its branches formed anastomoses with the buccal branch of the maxillary artery,creating a vascular network in the parotid and buccal regions.There was a vascular network around the buccal branch of the facial nerve,which was mostly small branches of the facial artery and the superficial temporal artery.Conclusion The buccal branch of the facial nerve exhibits a consistent anatomic relationship with the parotid duct and the facial artery.During the ligation of the facial artery,the parotid duct can serve as a landmark to accurately locate the buccal branch of the facial nerve,thereby significantly reducing the risk of inadvertent injury to the buccal branch of the facial nerve and the parotid duct.

This study established a droplet digital PCR(ddPCR)EvaGreen assay and probe methods for Schistosoma japonicum detection,and evaluated their application in detecting early infections in the S.japonicum host oncomelania and mice.Primers and corresponding probes for both ddPCR methods were designed and synthesized,and plasmids containing target sequences were constructed.The sensitivity of the two methods was tested through detection of the corresponding plasmids,and infectious and mixed oncomelania genomic DNA.Their specificity was evaluated by the detection of genomic DNA of negative oncomelania,Schistosoma mansoni,Clonorchis sinensis,Spirometra mansoni,and S.japonicum(as a positive control).The ddPCR probe method was evaluated by detection of early infection of oncomelania exposed tomiracidium with various ratios and incubation times,and the early migration and distribution of cercaria or schistosomula in mouse hosts infected with 200 cercaria via abdominal skin contact.According to standard curves constructed through the detection of plasmid serial dilutions,the regression equation for the EvaGreen assay was y=-0.839 9x+7.050 9,with a correlation coefficient R2=0.988 1,and the regression equation for the probe method was y=-1.047 5x+7.255 1,with a correlation coefficient R2=0.999 8.The lowest limit of plasmid detection with the probe method was between 38.94 cp/μL and 194.74 cp/μL.Both methods successfully detected positive reactions in the genomic DNA samples of infectious oncomelania at concentrations above 0.002 ng/μL and in the genomic DNA of each group of oncomelania mixtures.No significant differences between probe methods were observed in the detection values in the control group and the genomic DNA of negative oncomelania,S.mansoni,C.sinensis,and S.mansoni.However,the detection value of genomic DNA of negative oncomelania(291 ng/μL)with the EvaGreen assay was(20.3±4.39)cp/μL,a value significantly higher than the(1.5±0.1)cp/μL observed in the control group.For detection of early infection in oncomelania,the probe method detected Schistosoma japonicum DNA after 30 s incubation at room temperature with a≥5∶1 ratio of miracidium to oncomelania;the detection value peaked after a short time(5 min),and the peak value showed a fold increase similar to the increase in the miracidium to oncomelania ratio.Detection of early stage infection in mice with the probe method revealed that the schistosomula entered the lungs on day 2 and the liver on day 4,and continually migrated within the organs with abundant blood supply(spleen,kidney,and brain)in the first 9 days;moreover,a tendency toward ectopic parasitism was observed in the heart and pancreas on day 9,and a constantly negative control level was observed in the testes.The ddPCR probe method was more accurate and specific than the EvaGreen assay in the detection of plasmids,and infectious and mixed oncomelania,and the latter showed non-specific reactions in negative oncomelania detection.In a practical application,the probe method was demonstrated to be sensitive,to effectively reflect the early infection of oncomelania,and to reveal schistosomula migration and distribution in multiple organs of infected mice.

Objective To investigate the expression of serum microRNA(miR)-16a and mir-15b in patients with early-onset neonatal sepsis(EONS)and its value in clinical diagnosis and prognosis evaluation.Methods 114 children diagnosed with EONS admitted to the Department of Neonatology,Baoding Hospital of Beijing Children's Hospital Affiliated to Capital Medical University from October 2020 to October 2023 were selected as the EONS group,and 114 healthy newborns in the hospital during the same period were selected as the control group.Follow up was conducted on all EONS patients for a period of 6 months,including 34 cases in the poor prognosis group and 80 cases in the good prognosis group.The relative expression levels of miR-16a and miR-15b were measured within 6h of admission using real-time fluorescence quamtitative PCR(qRT-PCR).ROC curve was applied to analyze the diagnostic and prognostic value of serum miR-16a and miR-15b for EONS.Multivariate Logistic regression was applied to analyze the factors that affected the poor prognosis of EONS.Results Compared with the reference group,the birth weight of children in the EONS group(3.44±0.35kg/m2 vs 3.89±0.40kg/m2)was obviously reduced,and there was a obvious difference(35/114,1/114)in pathological jaundice between the two groups,and the differences were statistically significant(t=9.040,χ2=35.922,all P＜0.05).Compared with the reference group,the serum levels of miR-16a and miR-15b in the EONS group were obviously increased,and the differences was statistically significant(t=7.699,8.606,all P＜0.05).The area under the curve(AUC)of the ROC curve for the combined diagnosis of EONS by serum miR-16a and miR-15b was higher than that of miR-16a and miR-15b alone,and the differences were statistically significant(Z=2.619,2.157,all P<0.05).Compared with the good prognosis group,the poor prognosis group had a obvious decrease in body weight at birth,and a obvious increase in serum levels of miR-16a and miR-15b,and the differences were statistically significant(t=5.434,6.308,all P＜0.05).The AUC of serum miR-16a and miR-15b combined for prognostic evaluation was higher than that of miR-16a and miR-15b evaluated separately,and the differences were statistically significant(Z=2.364,2.073,all P＜0.05).Birth weight,miR-16a,miR-15b expression level was a prognostic factor of EONS(Wald χ2=33.459,16.146,49.797,all P<0.05).Conclusion MiR-16a and miR-15b are highly expressed in the serum of children with EONS,and they have certain diagnostic and prognostic value for EONS.