Liver fibrosis is the core pathological stage of the progression of various chronic liver diseases to liver cirrhosis， and hepatic stellate cell （HSC） activation and the abnormal accumulation of collagen fibers are important processes for the development and progression of liver fibrosis. In recent years， studies have shown that HSC activation is regulated by the complex interactions between various organelles （including mitochondria， endoplasmic reticulum， Golgi apparatus， lysosome， and peroxisomes）， and such interactions affect the key cellular processes such as energy metabolism， protein synthesis and folding， reactive oxygen species balance， and autophagy， thereby participating in the progression of liver fibrosis. Meanwhile， traditional Chinese medicine and its active ingredients with multi-target synergistic effects have attracted wide attention. From the perspective of the interaction between organelles， this article systematically elaborates on the specific mechanism of such interactions in the progression of liver fibrosis and reviews how traditional Chinese medicine inhibits HSC activation and collagen production by regulating the function of these organelle and their interaction networks， thereby exerting an anti-liver fibrosis effect， in order to provide a theoretical basis for in-depth understanding of the pathological mechanism of liver fibrosis and the development of new traditional Chinese medicine intervention strategies.

OBJECTIVE To investigate the effects of total flavonoids from Carthamus tinctorius L. （TFCTL） on hepatic stellate cell （HSC） activation based on the microRNA （miRNA）-204/NUAK family SNF1-like kinase 1 （NUAK1）/Hippo signaling axis， thereby elucidating the potential mechanism underlying their antifibrotic effects. METHODS The HSC-T6 cells were divided into control group， model group， TFCTL low-concentration group （20 μg/mL）， TFCTL medium-concentration group （40 μg/mL）， and TFCTL high-concentration group （60 μg/mL）. Except for control group， the remaining groups were treated with 5 ng/mL of transforming growth factor-β to induce the activation of hepatic stellate cells， followed by the addition of corresponding drug solutions/culture medium and incubation for 24 hours. Cell apoptosis was assessed， the expression levels of α-smooth muscle actin （α-SMA）， type Ⅰ collagen （Collagen Ⅰ） and proteins associated with the Hippo/Yes-associated protein （YAP） pathway [YAP， large tumor suppressor kinase 1 （LATS1）， and mammalian STE20-like kinase 1 （MST1）] were detected. Additionally， cell transfection was used to investigate the activity of the miRNA-204/NUAK1/Hippo signaling axis at both the genetic and protein levels. RESULTS After intervention with TFCTL， the apoptosis rate of HSC-T6 cells and the protein expressions of MST1 （except for the TFCTL high-concentration group） and LATS1 were significantly increased （P＜0.05）， while the protein expressions of α-SMA， CollagenⅠ， and YAP （except for the TFCTL medium-concentration group） were significantly decreased （P＜0.05）. Further results from cell transfection experiments revealed that after transfection with miRNA-204 mimics， the mRNA it’s protein expressions of α-SMA， CollagenⅠ， NUAK1， and YAP in HSC-T6 cells were significantly decreased （P＜0.05）， while the mRNA and protein expressions of LATS1 and the mRNA expression of MST1 were significantly increased （P＜0.05）. Conversely， the results were opposite following transfection with miRNA-204 inhibitors. CONCLUSIONS TFCTL can exert anti-hepatic fibrosis effects by up-regulating the expression of miRNA-204， thereby down- regulating the expressions of NUAK1， inactivating the Hippo/YAP pathway， which in turn suppresses the activation of HSC and promotes their apoptosis.

Metabolic dysfunction-associated fatty liver disease （MAFLD） is a chronic metabolic liver disorder with complex etiologies. Different clinical phenotypes of MAFLD （such as obesity， hyperlipidemia， type 2 diabetes mellitus， the postmenopausal state， and chronic hepatitis B） have different mechanisms of action in the development and progression of MAFLD， leading to high heterogeneity in its clinical progression and prognosis. This article systematically reviews the pathogeneses and clinical features of the above five clinical phenotypes of MAFLD and elaborates on the corresponding individualized diagnosis and treatment regimens integrating traditional Chinese medicine and Western medicine， in order to provide a reference for clinical practice and improve clinical diagnosis and treatment.

OBJECTIVE: To explore the incidence, clinical features, genetic variant characteristics and prognosis of Glutaric acidemia type I (GA1) among neonates from Henan Province. METHODS: A total of 814 625 neonates undergoing screening for inherited metabolic diseases by tandem mass spectrometry (MS/MS) at the Third Affiliated Hospital of Zhengzhou University from January 2016 to December 2022 were selected as the study subjects. A retrospective method was adopted to collect the clinical data of the patients. Whole exome sequencing was carried out to detect GCDH gene variants in individuals with positive results by GA1 newborn screening, and Sanger sequencing was used to verify the candidate variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the pathogenicity of candidate variants was rated. This study was approved by the Medical Ethics Committee of the Hospital (Ethics Number: 2019 Medical Ethics Review No. 67). RESULTS: Eight cases of GA1 were diagnosed among the 814 625 neonates. Blood glutaryl carnitine (C5DC) and urine glutaric acid (GA) levels of the 8 children were higher than the normal reference values. In total 12 variants were detected, all of which were missense variants. c.1064G>A (p.Arg355His) was the most common one, accounting for 21.4% (3/14). Three GCDH gene variants, including 1297G>C (p.Ala433Pro), c.467G>A (p.Gly156Asp) and c.1125T>G (p.Cys375Trp), were previously unreported. REVEL software analysis predicted that all of the three variants were harmful. 3D protein structure modeling indicated that the three variants may cause amino acid residue alterations, and c.1297G>C (p.Ala433Pro) and c.1125T>G (p.Cys375Trp) may result in increase in hydrogen bonds and affect the function of GCDH protein. By December 2023, one of the eight children had deceased, and another child had severe clinical symptoms with poor prognosis. Six children had a good prognosis, of which two had mild motor development delay and four had normal development without clinical symptoms. CONCLUSION The incidence of GA1 in newborns screened by MS/MS in Henan Province is 1/101 828, and the carrier rate of pathogenic GCDH variants is 1/160. The c.1064G>A (p.Arg355His) may be the hotspot variant of the GCDH gene among children with GA1 in Henan. Discovery of the three novel variants has enriched the mutational spectrum of the GCDH gene and provide a basis for the early diagnosis, treatment, prognosis and genetic counseling of this disease.
Humans ; Amino Acid Metabolism, Inborn Errors/epidemiology* ; Glutaryl-CoA Dehydrogenase/chemistry* ; Infant, Newborn ; Female ; Neonatal Screening/methods* ; Male ; Brain Diseases, Metabolic/epidemiology* ; China/epidemiology* ; Retrospective Studies ; Mutation ; Genetic Variation ; Glutarates

Objective To explore the effect on the expression of ERα/PGC1α/PPARα pathway,the therapeutic effect of Danhe Liuwei Dihuang Decoction on ovariectomized NAFLD rats was observed.Methods 60 female non-pregnant SPF SD rats were randomly divided into sham operation group(Sham),model group(Model),estrogen group(E2),traditional Chinese medicine high,medium and low dose group(DHR-H/M/L).Sham group was given normal diet after sham operation,and the other groups were given high-fat,high-fructose and high-cholesterol diet(HF/HF/HC)after bilateral ovariectomy to prepare postmenopausal NAFLD model.The Sham group and the Model group were given normal saline by gavage,and the other groups were given corresponding doses of drugs by gavage.The intervention time was 12 weeks,and the experimental period was 14 weeks.The general condition and body weight of rats in each group were recorded.HE staining of uterus was used to observe the morphology of uterus,HE staining of liver and oil red O staining were used to observe steatosis.Serum liver function(ALT,AST)and lipid metabolism indexes(TG,TC)were measured by automatic biochemical analyzer.The levels of serum E2 and free fatty acid(FFA)and the expression of FFA and TG in liver tissue were detected by ELISA.The mRNA and protein expressions of ERα,PGC1α and PPARα in liver tissues were detected by RT-PCR and Western blot.Results Compared with the Model group,by Danhe Liuwei Dihuang Decoction,the body weight and liver weight of ovariectomized NAFLD rats significantly decreased,liver fat deposition significantly decreased,E2 level increased,ALT,AST,TG,TC,FFA and liver tissue homogenate FFA,TG content significantly decreased,liver ERα,PGC1α,PPARα mRNA and protein expression up-regulated(P<0.05 or P<0.01).Conclusion Danhe Liuwei Dihuang Decoction can improve liver steatosis in postmenopausal NAFLD model rats.The mechanism may be related to up-regulating the expression of ERα/PGC1α/PPARα signaling pathway to promote liver FFA oxidation and reduce liver TG deposition.

Objective To retrieve,evaluate and summarize the best evidence for blood glucose monitoring and management in patients with diabetes of the exocrine pancreas(DEP),and to provide evidence-based basis for clinical practice.Methods According to the"6S"evidence model,relevant evidence on blood glucose management in patients with DEP was searched from computer decision support systems,guideline networks,professional association websites and databases from top to bottom,covering clinical decision-making,clinical practice guidelines,evidence summaries,systematic reviews,expert consensuses,and randomized controlled trials.The search scope was from the establishment of the databases to June 30,2024.Totally 2 researchers independently evaluated the quality of the included literature,and extracted data and summarized evidence that met the criteria.Results A total of 10 articles were included,including 3 guidelines,1 expert consensus,2 systematic reviews,l clinical decision,1 RCT,and 2 cohort studies.Finally,26 pieces of best evidence were formed from 5 aspects,including blood glucose management team construction and goals,blood glucose monitoring methods,glucose control medication management,lifestyle health education and treatment of hypoglycemia.Conclusion This study summarizes the best evidence for blood glucose monitoring and management in DEP patients,which can provide resource preparation for clinical translation and a basis for medical staff to carry out blood glucose management in DEP patients.

AIM:To examine the impact of An-gong Niuhuang pill in conjunction with convention-al therapy on the recovery of neurological function in patients with cerebral hemorrhage(ICH)charac-terized by phlegm-heat internal obstruction,and to evaluate its efficacy in enhancing neurological func-tion,promoting brain tissue repair,and mitigating oxidative stress.Additionally,this study aims to elu-cidate the potential mechanisms underlying these effects through the AMPK-PGC1α-Nrf2 pathway.METHODS:A total of 130 patients with ICH charac-terized by phlegm-heat internal closure were re-cruited and randomly allocated into a control group(receiving conventional treatment)and a study group(receiving Angong Niuhuang pill in combination with conventional treatment)using the random number table method,with 65 cases in each group.Post-treatment assessments included the National Institutes of Health Stroke Scale(NI-HSS)score,Glasgow Coma Scale(GCS)score,modi-fied Rankin scale(mRS)score,as well as tissue lev-els of AMP-activated protein kinase(AMPK),peroxi-some proliferator-activated receptor gamma coacti-vator-1α(PGC1α),and nuclear factor erythroid 2-related factor 2(NRF2).Additionally,serum levels of superoxide dismutase(SOD),malondialdehyde(MDA),reactive oxygen species(ROS),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),inter-leukin-6(IL-6),glutathione peroxidase 4(GPX4),ac-yl-coenzyme A synthetase long-chain family mem-ber 4(ACSL4),iron ions,tumor volume,brain ede-ma volume,and the incidence of adverse reactions were evaluated.RESULTS:Each group comprised 65 cases.Post-treatment,the NIHSS score(9.97±1.22)and mRS score(2.29±0.33)in the Research Group were significantly lower than those in the Control Group(11.17±1.52 and 2.64±0.45,respec-tively),with p-values less than 0.05.Additionally,the GCS score for the Research Group(14.57±1.19)was notably higher compared to the control group(13.18±1.05)(P＜0.05).After treatment,the expres-sions of AMPK[(3.34±0.81)ng/mL],PGC1α[(2.30±0.67)ng/mL]and NRF2[(3.72±0.85)ng/mL]in the tissues of the group were higher than those of the control group[(2.63±0.65)ng/mL,(1.83±0.70)ng/mL,(2.91±0.96)ng/mL](P＜0.05).After treatment,the levels of serum SOD[(98.76±6.67)U/mL]and IL-10[(40.37±5.61)pg/mL]in the research group were higher than those in the Control group group[(89.65±6.89)U/mL,(35.69±4.65)pg/mL](P＜0.05);the serum levels of MDA[(3.36±0.62)nmol/mL],ROS[(126.35±23.74)U/mL],TNF-α[(17.22±2.07)pg/mL]and IL-6[(33.37±3.76)pg/mL]in the group were lower than those in the control group group[(3.76±0.83)nmol/mL,(159.85±26.67)nmol/mL,(19.15±2.34)nmol/mL,(41.26±4.91)nmol/mL](P＜0.05).After treatment,the serum GPX4 level in the Research Group[(9.87±1.25)ng/mL]was higher than that in the control group[(8.16±1.12)ng/mL](P＜0.05).The serum levels of ACSL4[(8.74±1.45)ng/mL]and iron ion[(27.44±3.35)μmol/L]in the study group were lower than those in the control group(10.12±2.11)ng/mL,(30.46±3.17)μmol/L](P＜0.05).After treatment,the hematoma volume[(6.46±1.13)mL]and brain edema volume[(11.47±1.76)mL]in the research group were bet-ter than those in the Control group[(8.71±1.02)mL,(3.41±2.04)mL](P＜0.05).CONCLUSION:The combination of Angong Niuhuang pill with conven-tional treatment demonstrates significant efficacy in improving neurological function,promoting brain tissue repair,and alleviating oxidative stress,thereby exhibiting high application value in clinical practice.

Objective To explore the role of secreted phosphoprotein 1(SPP1)and myeloid differentiation primary response 88(MYD88)in morphine-induced cardiomyocyte apoptosis.Methods A morphine addiction model was established in Sprague-Dawley(SD)rats.Twelve SD rats were randomly assigned to the normal saline(NS)group or the morphine-dependent(DAM)group.Histopathological analysis was employed to observe and compare myocardial tissue morphology between the two groups.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was performed to assess the number of apoptotic cells in each group.The expression levels of SPP1 and MYD88 were evaluated using immunohistochemistry.Quantitative real-time poly merase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression of SPP1,MYD88,Bax,Bcl2,Caspase-3,and Caspase-9.Simultaneously,Western blot analysis was used to detected the expression of Cleaved Caspase-3 and Cleaved Caspase-9 proteins.In vitro,SPP1 expression was knocked down in primary neonatal rat cardiomyocytes(NRCMs),and cells were divided into three groups:control(CON),morphine treated(DA),and shSPP1#3+DA.Cell viability was assessed using the CCK-8 assay,and apoptosis rates were determined by flow cytometry.Results HE and TUNEL staining of myocardial tissues from morphine-addicted SD rats revealed that,compared with the NS group,myofibrils in the DAM group exhibited partial disruption and a significant increase in apoptotic cells(P<0.05).Western blot and RT-qPCR analyses demonstrated that,relative to the NS group,the mRNA and protein levels of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 were significantly upregulated in the DAM group(P<0.05),whereas Bcl2 expression was significantly downregulated at both mRNA and protein levels(P<0.05),and the protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were also increased.with all differences being statistically significant.In NRCMs following morphine intervention,cell viability in the DA group was markedly reduced compared to the CON group(P<0.05),accompanied by a signifi-cant increase in apoptosis rate(P<0.05).Consistently,Western blot and RT-qPCR results showed elevated mRNA and protein expression of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 in the DA group(P<0.05),along with decreased Bcl2 expression(P<0.05).The protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were elevated simultaneously.In contrast,the shSPP1#3+DA group exhibited opposing trends compared to the DA group,with statistically sig nificant differences(P<0.05).Conclusion SPP1 and MYD88 play critical roles in mediating morphine-induced cardiomyocyte apoptosis,and silencing SPP1 has been shown to significantly reduce the extent of cardiomyocyte apoptosis following morphine exposure.

AIM:To examine the impact of An-gong Niuhuang pill in conjunction with convention-al therapy on the recovery of neurological function in patients with cerebral hemorrhage(ICH)charac-terized by phlegm-heat internal obstruction,and to evaluate its efficacy in enhancing neurological func-tion,promoting brain tissue repair,and mitigating oxidative stress.Additionally,this study aims to elu-cidate the potential mechanisms underlying these effects through the AMPK-PGC1α-Nrf2 pathway.METHODS:A total of 130 patients with ICH charac-terized by phlegm-heat internal closure were re-cruited and randomly allocated into a control group(receiving conventional treatment)and a study group(receiving Angong Niuhuang pill in combination with conventional treatment)using the random number table method,with 65 cases in each group.Post-treatment assessments included the National Institutes of Health Stroke Scale(NI-HSS)score,Glasgow Coma Scale(GCS)score,modi-fied Rankin scale(mRS)score,as well as tissue lev-els of AMP-activated protein kinase(AMPK),peroxi-some proliferator-activated receptor gamma coacti-vator-1α(PGC1α),and nuclear factor erythroid 2-related factor 2(NRF2).Additionally,serum levels of superoxide dismutase(SOD),malondialdehyde(MDA),reactive oxygen species(ROS),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),inter-leukin-6(IL-6),glutathione peroxidase 4(GPX4),ac-yl-coenzyme A synthetase long-chain family mem-ber 4(ACSL4),iron ions,tumor volume,brain ede-ma volume,and the incidence of adverse reactions were evaluated.RESULTS:Each group comprised 65 cases.Post-treatment,the NIHSS score(9.97±1.22)and mRS score(2.29±0.33)in the Research Group were significantly lower than those in the Control Group(11.17±1.52 and 2.64±0.45,respec-tively),with p-values less than 0.05.Additionally,the GCS score for the Research Group(14.57±1.19)was notably higher compared to the control group(13.18±1.05)(P＜0.05).After treatment,the expres-sions of AMPK[(3.34±0.81)ng/mL],PGC1α[(2.30±0.67)ng/mL]and NRF2[(3.72±0.85)ng/mL]in the tissues of the group were higher than those of the control group[(2.63±0.65)ng/mL,(1.83±0.70)ng/mL,(2.91±0.96)ng/mL](P＜0.05).After treatment,the levels of serum SOD[(98.76±6.67)U/mL]and IL-10[(40.37±5.61)pg/mL]in the research group were higher than those in the Control group group[(89.65±6.89)U/mL,(35.69±4.65)pg/mL](P＜0.05);the serum levels of MDA[(3.36±0.62)nmol/mL],ROS[(126.35±23.74)U/mL],TNF-α[(17.22±2.07)pg/mL]and IL-6[(33.37±3.76)pg/mL]in the group were lower than those in the control group group[(3.76±0.83)nmol/mL,(159.85±26.67)nmol/mL,(19.15±2.34)nmol/mL,(41.26±4.91)nmol/mL](P＜0.05).After treatment,the serum GPX4 level in the Research Group[(9.87±1.25)ng/mL]was higher than that in the control group[(8.16±1.12)ng/mL](P＜0.05).The serum levels of ACSL4[(8.74±1.45)ng/mL]and iron ion[(27.44±3.35)μmol/L]in the study group were lower than those in the control group(10.12±2.11)ng/mL,(30.46±3.17)μmol/L](P＜0.05).After treatment,the hematoma volume[(6.46±1.13)mL]and brain edema volume[(11.47±1.76)mL]in the research group were bet-ter than those in the Control group[(8.71±1.02)mL,(3.41±2.04)mL](P＜0.05).CONCLUSION:The combination of Angong Niuhuang pill with conven-tional treatment demonstrates significant efficacy in improving neurological function,promoting brain tissue repair,and alleviating oxidative stress,thereby exhibiting high application value in clinical practice.

Objective To explore the role of secreted phosphoprotein 1(SPP1)and myeloid differentiation primary response 88(MYD88)in morphine-induced cardiomyocyte apoptosis.Methods A morphine addiction model was established in Sprague-Dawley(SD)rats.Twelve SD rats were randomly assigned to the normal saline(NS)group or the morphine-dependent(DAM)group.Histopathological analysis was employed to observe and compare myocardial tissue morphology between the two groups.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was performed to assess the number of apoptotic cells in each group.The expression levels of SPP1 and MYD88 were evaluated using immunohistochemistry.Quantitative real-time poly merase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression of SPP1,MYD88,Bax,Bcl2,Caspase-3,and Caspase-9.Simultaneously,Western blot analysis was used to detected the expression of Cleaved Caspase-3 and Cleaved Caspase-9 proteins.In vitro,SPP1 expression was knocked down in primary neonatal rat cardiomyocytes(NRCMs),and cells were divided into three groups:control(CON),morphine treated(DA),and shSPP1#3+DA.Cell viability was assessed using the CCK-8 assay,and apoptosis rates were determined by flow cytometry.Results HE and TUNEL staining of myocardial tissues from morphine-addicted SD rats revealed that,compared with the NS group,myofibrils in the DAM group exhibited partial disruption and a significant increase in apoptotic cells(P<0.05).Western blot and RT-qPCR analyses demonstrated that,relative to the NS group,the mRNA and protein levels of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 were significantly upregulated in the DAM group(P<0.05),whereas Bcl2 expression was significantly downregulated at both mRNA and protein levels(P<0.05),and the protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were also increased.with all differences being statistically significant.In NRCMs following morphine intervention,cell viability in the DA group was markedly reduced compared to the CON group(P<0.05),accompanied by a signifi-cant increase in apoptosis rate(P<0.05).Consistently,Western blot and RT-qPCR results showed elevated mRNA and protein expression of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 in the DA group(P<0.05),along with decreased Bcl2 expression(P<0.05).The protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were elevated simultaneously.In contrast,the shSPP1#3+DA group exhibited opposing trends compared to the DA group,with statistically sig nificant differences(P<0.05).Conclusion SPP1 and MYD88 play critical roles in mediating morphine-induced cardiomyocyte apoptosis,and silencing SPP1 has been shown to significantly reduce the extent of cardiomyocyte apoptosis following morphine exposure.