Developing and identifying effective medications and targets for treating hepatic fibrosis is an urgent priority. Our previous research demonstrated the efficacy of artesunate (ART) in alleviating liver fibrosis by eliminating activated hepatic stellate cells (HSCs). However, the underlying mechanism remains unclear despite these findings. Notably, endocytic adaptor protein (NUMB) has significant implications for treating hepatic diseases, but current research primarily focuses on liver regeneration and hepatocellular carcinoma. The precise function of NUMB in liver fibrosis, particularly its ability to regulate HSCs, requires further investigation. This study aims to elucidate the role of NUMB in the anti-hepatic fibrosis action of ART in HSCs. We observed that the expression level of NUMB significantly decreased in activated HSCs compared to quiescent HSCs, exhibiting a negative correlation with the progression of liver fibrosis. Additionally, ART induced senescence in activated HSCs through the NUMB/P53 tumor suppressor (P53) axis. We identified NUMB as a crucial regulator of senescence in activated HSCs and as a mediator of ART in determining cell fate. This research examines the specific target of ART in eliminating activated HSCs, providing both theoretical and experimental evidence for the treatment of liver fibrosis.
Hepatic Stellate Cells/cytology* ; Liver Cirrhosis/genetics* ; Artesunate/pharmacology* ; Cellular Senescence/drug effects* ; Membrane Proteins/genetics* ; Animals ; Humans ; Nerve Tissue Proteins/genetics* ; Tumor Suppressor Protein p53/genetics* ; Male ; Mice

Objective To investigate the mechanisms that mediate the neuroprotective effect of the intestinal microbial metabolite sodium butyrate(NaB)in a mouse model of Parkinson's disease(PD)via the gut-brain axis.Methods Thirty-nine 7-week-old male C57BL/6J mice were randomized equally into control group,PD model group,and NaB treatment group.In the latter two groups,PD models were established by intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)once daily for 5 consecutive days,and normal saline was injected in the control group.After modeling,the mice received daily gavage of NaB(300 mg/kg)or an equal volume of saline for 14 days.Behavioral tests were carried out to assess the changes in motor function of the mice,and Western blotting was performed to detect the expressions of tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the striatum and nuclear factor-κB(NF-κB),tumor necrosis factor(TNF-α),interleukin 6(IL-6),and the tight junction proteins ZO-1,Occludin,and Claudinin the colon.HE staining was used to observe inflammatory cell infiltration in the colon of the mice.RNA sequencing analysis was performed to identify the differentially expressed genes in mouse colon tissues,and their expressions were verified using qRT-PCR and Western blotting.Results The mouse models of PD with NaB treatment showed significantly increased movement speed and pulling strength of the limbs with obviously upregulated expressions of TH,Occludin,and Claudin and downregulated expressions of α-syn,NF-κB,TNF-α,and IL-6(all P<0.05).HE staining showed that NaB treatment significantly ameliorated inflammatory cell infiltration in the colon of the PD mice.RNA sequencing suggested that Bmal1 gene probably mediated the neuroprotective effect of NaB in PD mice(P<0.05).Conclusion NaB can improve motor dysfunction,reduce dopaminergic neuron loss in the striatum,and ameliorate colonic inflammation in PD mice possibly through a mechanism involving Bmal1.

OBJECTIVE To explore the anti-liver fibrosis effect and mechanism of Atractylenolide Ⅲ-induced hepatic stellate cell(HSC)senescence.METHODS ASCT2 siRNA and Atractylenolide Ⅲ(40 μmol·L-1)acted on human hepatic stellate cells LX2 respectively to inhibit ASCT2,MTT was used to evaluate cell viability,EdU method was used to detect cell proliferation,and se-nescence associated-β-galactosidase(SA-β-Gal)staining was used to detect cell senescence;Western blot was used to detect chan-ges in the LC3-Ⅱ/Ⅰ ratio in LX2 cells,laser confocal detection was used to detect changes in LC3 autophagy flow and error protein accumulation,and the fluorescence of the lysosomal marker LAMP1 was also observed to detect lysosomal function and quantity;kits were applied to detect ROS and MDA levels as well as SOD activity in LX2 cells,and flow cytometry was used to analyze mitochondrial ROS levels and membrane potential.A CCl4-induced mouse liver fibrosis model was constructed.Atractylenolide Ⅲ was administered at 20,30,or 40 mg·kg-1.HE,Masson,and Sirius Red staining were used to observe liver tissue damage and collagen deposition.Western blot was used to detect the expression levels of P21 and P16 in mice in each group,and SA-β-Gal staining and immunohistochemistry were used to analyze the situation and origin of senescent cells.RESULTS After inhibiting ASCT2,the viabil-ity of LX2 cells decreased and senescence increased(P<0.01).Meanwhile,the autophagy function was enhanced and the number of lysosomes was increased but the function was weakened.After adding chloroquine(CQ)to clear lysosomes,the cell viability and auto-phagy function increased(P<0.01).After inhibiting ASCT2,the levels of MDA and ROS in LX2 cells increased,and the activity of SOD decreased(P<0.01).Among them,the level of mitochondrial ROS increased and the membrane potential decreased(P<0.01).After adding rotenone,the cellular redox homeostasis was improved,and the number of lysosomes was restored(P<0.01).In vivo experimental results showed that compared with the model group,Atractylenolide Ⅲ improved liver tissue structural damage and collagen deposition,induced HSC senescence in liver tissue of mice with liver fibrosis,and inhibited HSC activation marker α-smooth muscle actin(α-SMA),promoted the expression of senescence indicators P16 and P21(P<0.01).CONCLUSION Atractylenol-ide Ⅲ induces an increase in mitochondrial ROS and a decrease in membrane potential by inhibiting ASCT2,which further promotes the enhancement of HSC autophagy function,increases the number of lysosomes and weakens their function,thereby inducing the se-nescence of activated HSCs.

Objective To investigate the mechanisms that mediate the neuroprotective effect of the intestinal microbial metabolite sodium butyrate(NaB)in a mouse model of Parkinson's disease(PD)via the gut-brain axis.Methods Thirty-nine 7-week-old male C57BL/6J mice were randomized equally into control group,PD model group,and NaB treatment group.In the latter two groups,PD models were established by intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)once daily for 5 consecutive days,and normal saline was injected in the control group.After modeling,the mice received daily gavage of NaB(300 mg/kg)or an equal volume of saline for 14 days.Behavioral tests were carried out to assess the changes in motor function of the mice,and Western blotting was performed to detect the expressions of tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the striatum and nuclear factor-κB(NF-κB),tumor necrosis factor(TNF-α),interleukin 6(IL-6),and the tight junction proteins ZO-1,Occludin,and Claudinin the colon.HE staining was used to observe inflammatory cell infiltration in the colon of the mice.RNA sequencing analysis was performed to identify the differentially expressed genes in mouse colon tissues,and their expressions were verified using qRT-PCR and Western blotting.Results The mouse models of PD with NaB treatment showed significantly increased movement speed and pulling strength of the limbs with obviously upregulated expressions of TH,Occludin,and Claudin and downregulated expressions of α-syn,NF-κB,TNF-α,and IL-6(all P<0.05).HE staining showed that NaB treatment significantly ameliorated inflammatory cell infiltration in the colon of the PD mice.RNA sequencing suggested that Bmal1 gene probably mediated the neuroprotective effect of NaB in PD mice(P<0.05).Conclusion NaB can improve motor dysfunction,reduce dopaminergic neuron loss in the striatum,and ameliorate colonic inflammation in PD mice possibly through a mechanism involving Bmal1.

Objective:To correlate serum homocysteine (Hcy) level with psychiatric symptoms and social ability in patients with schizophrenia.Methods:A total of 143 patients with schizophrenia who received treatment in the Third Hospital of Quzhou from January 2018 to January 2020 were included in this study. They were randomly divided into abnormal Hcy group (Hcy ≥ 10) μmol/L, n = 46) and normal Hcy group (Hcy < 10 μmol/L, n = 97) according to different serum Hcy levels. Baseline data, the scores of the Brief Psychiatric Rating Scale (BPRS), the Positive and Negative Syndrome Scale (PANSS), and the Substance Dependence Severity Scale (SDSS) were compared between the two groups. The correlation between serum Hcy level and psychiatric symptoms and social ability was analyzed using the Pearson correlation analysis. Results:PANSS total score and BPRS total score were (77.86 ± 8.37) points and (94.47 ± 9.36) points, respectively in the abnormal Hcy group, and (74.37 ± 7.94) points and (90.35 ± 9.26) points, respectively in the normal Hcy group. There were significant differences in PANSS total score and BPRS total score between the two groups ( t = 2.41, 2.47, both P < 0.05). SDSS total score was significantly higher in the abnormal Hcy group than in the normal Hcy group [(11.75 ± 2.38) points vs. (10.53 ± 2.28) points, t = 2.88, P < 0.05]. Pearson correlation analysis showed that serum Hcy level was positively correlated with PANSS positive subscale score ( r = 0.73, P < 0.001), general mental symptom score ( r = 0.43, P = 0.032) and PANSS total score ( r = 0.53, P = 0.027), but it was not correlated with PANSS negative symptom score ( P > 0.05). Serum Hcy level was positively correlated with BPRS excitement subscale score ( r = 0.42, P = 0.037) and hostility subscale score ( r = 0.37, P = 0.047), but it was not correlated with anxiety, blunted affect, unusual thought content subscale scores and BPRS total score (all P > 0.05). Conclusion:Serum Hcy level is correlated with psychiatric symptoms and social ability in patients with schizophrenia. The higher the level of Hcy, the more severe the psychiatric symptoms, and the higher the degree of social disability.

Senescence of activated hepatic stellate cells (aHSCs) is a stable growth arrest that is implicated in liver fibrosis regression. Senescent cells often accompanied by a multi-faceted senescence-associated secretory phenotype (SASP). But little is known about how alanine-serine-cysteine transporter type-2 (ASCT2), a high affinity glutamine transporter, affects HSC senescence and SASP during liver fibrosis. Here, we identified ASCT2 is mainly elevated in aHSCs and positively correlated with liver fibrosis in human and mouse fibrotic livers. We first discovered ASCT2 inhibition induced HSCs to senescence in vitro and in vivo. The proinflammatory SASP were restricted by ASCT2 inhibition at senescence initiation to prevent paracrine migration. Mechanically, ASCT2 was a direct target of glutaminolysis-dependent proinflammatory SASP, interfering IL-1α/NF-κB feedback loop via interacting with precursor IL-1α at Lys82. From a translational perspective, atractylenolide III is identified as ASCT2 inhibitor through directly bound to Asn230 of ASCT2. The presence of -OH group in atractylenolide III is suggested to be favorable for the inhibition of ASCT2. Importantly, atractylenolide III could be utilized to treat liver fibrosis mice. Taken together, ASCT2 controlled HSC senescence while modifying the proinflammatory SASP. Targeting ASCT2 by atractylenolide III could be a therapeutic candidate for liver fibrosis.

Objective: To explore the value of bioelectrical impedance analysis in evaluating hemodialysis patients′ nutritional status. Methods: Cross-sectional design was employed to evaluate 193 hemodialysis patients′ nutritional status in a Level 3 hospital in Zhuhai, with bioelectrical impedance analysis, modified quantitative subjective global assessment, physical and biochemical measurement. The correlation between each index and the predictive effect of bioelectrical impedance analysis on malnutrition of patients were analyzed. Results: According to bioelectrical impedance analysis, hemodialysis patients′ average muscle mass was (44.06±8.84) kg, body fat rate was (19.81±8.89)%, fat free mass index was (17.70±2.45) kg/m², presumed bone mass was (2.49±0.46) kg, and extracellular water rate was (40.54±3.75)%. The bioelectrical impedance analysis indexes were significantly correlated with most other nutritional evaluation indexes (P<0.05). The muscle mass and bone mass of the malnourished group were lower than that of the well-nourished group (P<0.01). Multivariable logistic regression analysis found that body fat rate (OR=0.94, 95%CI 0.89-0.99), fat free body mass index (OR=0.79, 95%CI 0.66-0.93) were associated with lower risk of malnutrition, age (OR=1.08, 95% CI 1.04-1.12), dialysis age (OR=1.02, 95%CI 1.01-1.04), high sensitive C reactive protein (OR=1.11, 95%CI 1.03-1.23) were associated with higher risk of malnutrition. Conclusion Bioelectrical impedance analysis is a valid measurement on evaluating maintenance hemodialysis patients′ nutritional status. There has a high prevalence of malnutrition in hemodialysis patients. The lower risk of malnutrition is associated with higher body fat rate and higher fat free mass index, and the higher risk of malnutrition is associated with older age, longer dialysis age and higher level of high sensitive C reactive protein.

Objective To investigate the kidney function changes and the risk factors in elderly patients aged 80 years and over.Methods A total of 1054 hospitalized patients aged ≥80 years in our hospital were selected.Serum levels of creatinine,urea,uric acid and urinate were tested.Cockcroft-Gault equation was used to estimate glomerular filtrate rate (GFR).Results The renal function in 87.1 ％ of the patients was reduced.The percentage of patients with 30≤eGFR ＜60 ml · min-1 · 1.73 m-2 was 66.2％.The percentage of patients with 15≤eGFR ＜30 ml · min-1 · 1.73 · m-2had a gender difference (P＜0.05).Serum creatinine level was not an ideal indicator for early renal dysfunction in the elderly,especially in emaciated elderly people.Multivariate logistic regression analysis showed that gender,hypertension,coronary heart disease and hyperuricemia were independent risk factors for the decline of renal function (OR=1.937,1.602,1.842,7.020,respectively,all P＜0.05).Conclusions The rate of renal dysfunction is high in very old people.Many risk factors affect renal dysfunction.

Objective To evaluate the changes of IL-1β and IL-lra in hippocampus of rats with frequent febrile seizures(FS).Methods 21-day-old male Sprague-Dawley(SD)rats were randomly divided into 3 groups:normal control group(NC group),hyperthermic control group(HC group)and febrile seizures group(FS group).IL-1β and IL-1ra levels in hippocampus of these rats were determined by ELISA methods,IL-1β mRNA and IL-1ra mRNA levels in hippocampus were measured by RT-PCR.Results IL-1β mRNA and protein levels in hippocampus of rats in FS group were significantly higher than those in NC group and HC group(P＜0.01,P＜0.05).IL-1ra mRNA and protein levels in hippocampus of rats in FS group were significantly higher than those in NC group and HC group(P＜0.01,P＜0.05),too.IL-1ra/IL-1β protein ratio showed no significant difference among 3 groups.Conclusion Frequent FS can cause the increase of IL-1β and IL-lra mRNA and protein levels in hippocampus,suggesting that IL-1β and IL-1ra may be involved in the pathogenesis of hippocampal neuron damage in FS rat.

AIM:To investigate the protection mechanisms of rosiglitazone on diabetic nephropathy.METHODS:Rat mesangial cells(MC) were incubated in media containing 5.5 mmol/L normal control glucose,25 mmol/L high concentration glucose,25 mmol/L glucose +20 ?mol/L rosiglitazone maleate.Cells proliferation were assessed by CCK-8.Synthesis of fibronectin(FN),type Ⅳ collagen(Col-Ⅳ),transforming growth factor-?1(TGF-?1) and matrix metalloproteinase inhibitor-1(TIMP-1) in supernatant were determined by ELISA method,the activities of matrix metalloproteinase-2,9(MMP-2,9) in supernatant were determined by gelatinase zymography.RESULTS:Compared with control group,MC cultured with high concentration glucose showed a high growth rate and increased synthesis of Col-Ⅳand FN,decreased the activities of MMP-2 and MMP-9,and increased secretion of TGF-?1 and TIMP-1.Compared with high glucose group,these changes could be reversed by rosiglitazone intervention.CONCLUSION:Rosiglitazone could inhibit high concentration glucose-induced proliferation of mesangial cells,decrease synthesis of extracellular matrix,and increase degradation of extracellular matrix.