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.

Objective To investigate the prevalence, distribution characteristics, and influencing factors of mental health problems among the elderly, and to provide a scientific basis for policy-making.  Methods A convenience sampling method was used to investigate depression, anxiety, and cognitive function among permanent residents aged 65 and older at 59 mental health care sites for the elderly in Hubei Province. Multinomial logistic regression was employed to analyze influencing factors. Results The screening rates for depression, anxiety, and cognitive function at critical/high-risk levels among the elderly in Hubei Province were 9.7%, 5.4%, and 12.2%, respectively. Urban elderly had lower risks of depression and cognitive function at critical/high-risk levels compared to rural elderly (OR for critical depression = 0.640, P < 0.001; OR for high-risk depression = 0.595, P = 0.012; OR for critical cognitive function = 0.448, P < 0.001; OR for high-risk cognitive function = 0.188, P < 0.001). Six key population groups had higher risks of depression, anxiety, and cognitive function at critical/high-risk levels than others (OR for critical depression = 1.463, P < 0.001; OR for high-risk depression = 1.912, P < 0.001; OR for critical anxiety = 1.462, P < 0.001; OR for high-risk anxiety = 2.882, P < 0.001; OR for critical cognitive function = 1.381, P < 0.001; OR for high-risk cognitive function = 2.345, P < 0.001). A higher number of chronic diseases was associated with increased risks of critical and high-risk depression (OR for critical = 1.316, P < 0.001; OR for high-risk = 3.677, P < 0.001) and cognitive impairment (OR for critical depression = 1.316, P < 0.001; OR for high-risk depression = 3.677, P < 0.001; OR for critical anxiety = 1.512, P < 0.001; OR for high-risk anxiety = 1.801, P < 0.001). Conclusion It is recommended to expand mental health care sites in rural areas, improve the layout of mutual-support elderly care facilities, and explore sustainable models for rural elderly care. Efforts should also focus on enhancing social participation among the elderly through community-based activities, and strengthening cognitive screening and emotional regulation interventions, with particular attention to the mental health needs of older, isolated, and chronically ill individuals.

Objective To investigate the therapeutic effect and mechanism of MCC950 on benzalkonium chloride-in-duced dry eye(DED)in rats.Methods Among 60 male SD rats,50 rats were treated with 20 g·L-1 benzalkonium chlo-ride solution 3 times a day in both eyes to induce DED model,and a significant decrease in tear secretion was regarded as a successful modeling,and the remaining 10 rats were regarded as the control group.DED rats were randomly divided into 5 groups:DED,DED+10MCC950,DED+50MCC950,DED+100MCC950,and DED+500MCC950 groups,and were given 0.2 mL of 0,10,50,100,and 500 μmol·L-1 MCC950 in both eyes for two weeks,and then examined the amount of tear secretion,corneal histopathology(HE staining),reactive oxygen species(ROS),and inflammation factors(NLRP3,Caspase-1,IL-1β,IL-6).Results The tear secretion of rats in the DED group was significantly lower than that of the control group(P＜0.05).Except for the DED+10MCC950 group,the tear secretion of rats in different concentrations of MCC950 treatment groups was increased and positively correlated with the MCC950 dose(all P＜0.05).HE staining of the cornea showed that compared with the control group,the cornea of the DED group was significantly thinner,the arrange-ment of surface cells was disorganized,the number of cells was significantly reduced,and a large number of"vacuole"-like structures appeared;the corneal stroma was disorganized and sparsely structured.The degree of histopathological changes in the cornea of rats treated with different concentrations of MCC950 decreased with the increase of the concentration of MCC950.Compared with the control group,the relative expression level of ROS in the DED group of rats was significantly increased(P＜0.05).In each MCC950 treatment group,as the concentration of MCC950 increased,the relative expression level of corneal ROS staining in rats gradually decreased,showing a concentration-dependent effect(all P＜0.05).Com-pared with the control group,the levels of NLRP3,IL-1 β,Caspase-1 and IL-6 were significantly increased in the DED group(all P＜0.05).The levels of various inflammatory factors were dose-dependently reduced in the MCC950 treatment groups(all P＜0.05).Conclusion MCC950 can reduce oxidative stress and inflammation and improve the symptoms of benza-lkonium chloride-induced DED in rats by inhibiting the activation of NLRP3 inflammatory vesicles.

Periodontitis is a common oral disease caused by bacteria coupled with an excessive host immune response. Stem cell therapy can be a promising treatment strategy for periodontitis, but the relevant mechanism is complicated. This study aimed to explore the therapeutic potential of mitochondria from human embryonic stem cell-derived mesenchymal stem cells (hESC-MSCs) for the treatment of periodontitis. The gingival tissues of periodontitis patients are characterized by abnormal mitochondrial structure. Human gingival fibroblasts (HGFs) were exposed to 5 μg/mL lipopolysaccharide (LPS) for 24 h to establish a cell injury model. When treated with hESC-MSCs or mitochondria derived from hESC-MSCs, HGFs showed reduced expression of inflammatory genes, increased adenosine triphosphate (ATP) level, decreased reactive oxygen species (ROS) production, and enhanced mitochondrial function compared to the control. The average efficiency of isolated mitochondrial transfer by hESC-MSCs was determined to be 8.93%. Besides, a therapy of local mitochondrial injection in mice with LPS-induced periodontitis showed a reduction in inflammatory gene expression, as well as an increase in both the mitochondrial number and the aspect ratio in gingival tissues. In conclusion, our results indicate that mitochondria derived from hESC-MSCs can reduce the inflammatory response and improve mitochondrial function in HGFs, suggesting that the transfer of mitochondria between hESC-MSCs and HGFs serves as a potential mechanism underlying the therapeutic effect of stem cells.
Humans ; Gingiva/cytology* ; Fibroblasts/metabolism* ; Mitochondria/physiology* ; Mesenchymal Stem Cells/cytology* ; Animals ; Periodontitis/therapy* ; Mice ; Reactive Oxygen Species/metabolism* ; Inflammation ; Lipopolysaccharides ; Human Embryonic Stem Cells/cytology* ; Cells, Cultured ; Adenosine Triphosphate/metabolism* ; Male

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 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 (Approval 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 may 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.

[Objective]To explore the correlation between general anesthesia and mild cognitive impairment in older adults so as to provide new ideas for early prevention and timely intervention of mild cognitive impairment(MCI).[Methods]Based on the baseline survey of the Hubei memory and aging cohort study(2018-2023),the participants completed a thorough neuropsychological assessment and physical examination,and self-reported a history of general anesthesia and surgery.The association of general anesthesia and MCI in the elderly was analyzed using the logistic regression model.In addition,the stratification and interaction analysis of anesthesia history,anesthesia number and physical intellectual exercise were conducted separately.[Results]A total of 5 069 older adults aged 65 and above were included in this study,including 3 692 city dwellers and 1 377 rural people,among whom were 2 584 women(51%).Out of the 1 472 participants with history of general anesthesia,249 people(17.4%)had MCI.After controlling for confounding factors,there was a 39.6%increased risk of MCI in older adults who underwent general anesthesia[OR=1.396,95%CI(1.169,1.668),P<0.001],suggesting that general anesthesia may be an independent influence on MCI.For the older adults who had one general anesthesia[OR=1.235,95%CI(1.001,1.523),P=0.049],two general anesthesia[OR=1.779,95%CI(1.292,2.450),P<0.001],and three OR more general anesthesia[OR=2.395,95%CI(1.589,3.610),P<0.001],their risks of MCI were increased by 23.5%,77.9%,and 139.5%,respectively.Compared with the older adults without a history of general anesthesia who did not exercise,the risk of developing MCI was significantly negatively correlated with the exercise group,cognitive exercise group,and combined exercise and cognitive exercise groups(all P<0.001).The risk of developing MCI in the exercise group was 60.2%of that in the no exercise group[OR=0.602,95%CI(0.456,0.795)],the risk in the cognitive exercise group was 42.4%of that in the no exercise group[OR=0.424,95%CI(0.294,0.613)],and the risk in the combined exercise and cognitive exercise group was 27.0%of that in the no exercise group[OR=0.270,95%CI(0.208,0.353)].In the older adults with a history of general anesthesia,compared with the no exercise group,the risk of developing MCI was significantly negatively correlated with the cognitive exercise group and the combined exercise and cognitive exercise group(all P<0.05).The risk of developing MCI in the cognitive exercise group was 47.7%of that in the no exercise group[OR=0.477,95%CI(0.256,0.892)],the risk in the combined exercise and cognitive exercise group was 34.5%of that in the no exercise group[OR=0.345,95%CI(0.220,0.540)],while the risk in the exercise-only group did not show a significant difference.[Conclusion]The risk of MCI increased significantly in older adults with a history of general anesthesia,and this risk increased with the times of anesthesia.Physical and mental exercise reduces the risk of MCI.it is recommended that older adults with a history of anesthesia incorporate physical and mental exercise into their daily lives to prevent mild cognitive impairment.

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.

AIM:To investigate the efficacy and safety of checkpoint inhibitor lenvatinib combined with Tirelizumab in the treatment of advanced he-patocellular carcinoma(HCC).METHODS:A retro-spective analysis was performed on 52 patients with advanced HCC treated with Renvatinib,tirelli-zumab or their combination from January 2021 to December 2022 in Zhongda Hospital Affiliated to Southeast University,with disease progression,death and intolerance as endpoints.Efficacy was as-sessed according to RECIST 1.1 criteria and follow-up was conducted up to June 2023.The main end-points of the study were objective response rate(ORR),disease control rate(DCR),progression-free survival,PFS)and security.RESULTS:The combina-tion therapy significantly improved ORR and mPFS in patients compared with tirelizumab or lenvatinib monotherapy.Compared with single tirellizumab or lenvastinib,there was no significant difference in the adverse reactions associated with the combina-tion treatment,suggesting that the combination of the two was generally well tolerated and the side effects are controllable.CONCLUSION:Lonvastinib combined with tirelizumab is more effective in the treatment of HCC,can significantly prolong PFS,and is generally well tolerated,which may be a po-tential treatment for advanced HCC.

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.