Aim To evaluate the toxicity and mecha-nism of Epimedium sagittatum(Sieb.et Zucc.)Maxim aqueous extract(ESMAE)on HepaRG cells based on serum toxicology.Methods MTT assay was used to detect the activity of HepaRG cells after treatment with the serum containing ESMAE from SD rats.Western blot was used to detect the effects of the serum contai-ning drug on the expression of endoplasmic reticulum stress-related proteins(PERK,eIF-2α,ATF-4,GRP78,CHOP)and pyroptosis related proteins(NL-RP1,caspase-1,cleaved caspase-1,GSDMD,GSDMD-N).MTT assay was used to detect the activity of Hep-aRG cells after treatment with the liver homogenate containing ESMAE from SD rats.Results Twenty percent serum containing drug significantly decreased the viability of HepaRG cells,with the cells exhibiting swelling,rupture,and vesicle-like pyroptosis.The ex-pression levels of endoplasmic reticulum stress-related proteins PERK,eIF-2α,ATF-4,GRP78,and CHOP significantly increased.The expression levels of pro-teins involved in the NLRP1-mediated classical pyrop-tosis pathway were significantly increased.Finally the liver homogenate containing drug decreased the cell ac-tivity,and cells exhibited swelling,rupture,and vesicle-like pyroptosis.Conclusions After administration of ESMAE,the serum containing drug and the liver ho-mogenate containing drug of rats show toxicity to Hep-aRG cells,and can induce endoplasmic reticulum stress and activate the NLRP1/caspase-1/GSDMD pyroptosis pathway in HepaRG cells.

Aim To explore the role and mechanism of epimedokoreanin B(EKB)in HepaRG cell pyroptosis through endoplasmic reticulum stress and NLRP1-me-diated pyroptosis pathway.Methods The effect of EKB on the viability of HepaRG cells at different con-centrations was determined by MTT assay,and the cell growth status was recorded by Incucyte.Four groups of HepaRG cells were set up.The control group was cul-tured with complete medium for 24 h;the drug admin-istration group was cultured with three concentration gradients of 6.25,12.5 and 25 μmol·L-1 of EKB for 24 h.Western blot was used to detect the expression levels of endoplasmic reticulum stress-related proteins and pyroptosis-related proteins in the cells of each group.Results HepaRG cells showed cytotoxicity at a concentration of 6.25 μmol·L-1 for 24 h,and the half maximal inhibitory concentration(IC50)was 12.41 μmol·L-1.Incucyte recordings of the cell growth status showed that the cells in the control group were in good growth status,and the vesicular pyropto-sis cells appeared in the different concentrations of EKB and the cells swelled and ruptured after 24 h.Western blot showed that the protein expression levels of endoplasmic reticulum stress-related proteins pERK,eIF-2α,ATF-4,GRP78,and CHOP significantly in-creased in HepaRG cells at 25 μmol·L-1 of EKB compared with the control group.The proteins of the classical pathway of cellular pyroptosis mediated by NLRP1,caspase-1,cleaved caspase-1,GSDMD,GS-DMD-N significantly increased in HepaRG cells.Con-clusion EKB administration induces HepaRG cell py-roptosis,and EKB activates HepaRG cells to undergo endoplasmic reticulum stress and activates the NLRP1/caspase-1/GSDMD-mediated pyroptosis pathway.

Aim To study the difference in hepatotox-icity of psoralen on normal rats and Yin-deficiency rats from the perspective of lipid metabolism,so as to help explain the mechanism of psoralen cautiously used in patients with Yin deficiency recorded in ancient books.Methods SD rats were randomly divided into the nor-mal control group(carboxymethyl cellulose-Na,CMC-Na),normal administration group(CMC-Na+psor-alen),Yin-deficiency control group(CMC-Na+thy-roxine)and Yin-deficiency administration group(CMC-Na+thyroxine+psoralen).The model of Yin-deficiency was established by thyroxine(1 mg·kg-1)for ten days,and then psoralen(200 mg·kg-1)was given for three days.The serum indexes related to liver injury were detected by automatic biochemical analy-zer,the morphological changes of liver tissue were ob-served using HE and oil red O staining,and the relative transcription levels of lipid metabolism related enzymes and mRNA of transporter and endoplasmic reticulum stress related factors were detected using Real-time PCR.Results After intragastric administration of psoralen for three days,compared with the normal group,the levels of serum alanine aminotransferase(ALT),aspartate transaminase(AST),total bile acid(TBA)and triglyeride(TG)in Yin deficiency group increased more significantly,while TC,ALB and TP de-creased more significantly,and liver HE and oil red O staining showed more obvious lipid degeneration.TG synthesis factors adrenocortical carcinoma(ACC),fatty acid synthase(FASN)and sterolregulatory element binding protein-1(SREBP-1)were down-regulated more significantly,TG transport factors mili-total pro-tein(MTP)and lipoprotein pipase(LPL)were down-regulated more evidently,fatty acid β-oxidation related factors carnitine palmitoyltransferase 1A(CPT1A),carnitine/organic cation transporter 2(OCTN2)and peroxisome proliferators-activated receptors-alpha(PPARα)were down-regulated more apparently,TC transporter adenosine triphosphate binding cassette transporter G8(ABCG8)and bile acid receptor farne-soid X receptor(FXR)were down-regulated more ob-viously,and endoplasmic reticulum stress factor activa-ting transcription factor 4(ATF4)was up-regulated more significantly.Conclusions Psoralen can cause more severe hepatotoxicity in Yin deficiency rats than that in normal administration group,and its mechanism may be related to the disorder of hepatic lipid metabo-lism,aggravation of hepatic cholestasis and steatosis,and activation of endoplasmic reticulum stress re-sponse.

Objective To analyze the correlation between hemoglobin(Hb),anemia,and sarcopenia in the elderly population in Chongqing communities.Methods A cross-sectional study was conducted on elderly individuals who underwent healthy examinations at 5 community health service centers in Chongqing from March to August 2023.Demographic characteristics,social factors,body composition measurement,grip strength,6-meter gait speed and blood tests were assessed.Receiver operating characteristic(ROC)curve was utilized to evaluate the accuracy of Hb in predicting sarcopenia,and the Youden index was employed to determine the optimal Hb cut-off value for diagnosing sarcopenia and its components.Both unadjusted and adjusted logistic regression analyses were performed to examine the relationship between Hb and anemia with sarcopenia and its components.Results A total of 531 elderly populations were included,with an average age of(71.1±6.5)years.The overall prevalence of sarcopenia was 13.6%(72/531),including 29 males(40.3%)and 43 females(59.7%).Unadjusted analyses showed that Hb was correlated with sarcopenia,decreased muscle mass,slower gait speed,and reduced grip strength(P<0.05).After adjusting for all potential risk factors,Hb was still significantly associated with sarcopenia and reduced grip strength(P<0.05).For every 10 g/L increase in Hb,the risk of sarcopenia decreased by 2.3%,and the risk of reduced grip strength decreased by 1.7%(P<0.05).Anemia was correlated with sarcopenia,reduced muscle mass,and decreased grip strength in unadjusted analyses(P<0.05),while the correlation between anemia and reduced grip strength remained significant after adjustment for all potential risk factors(P<0.05).The optimal Hb cut-off value for diagnosing sarcopenia in males and females were 148 g/L and 128 g/L,respectively.Conclusions Hb is an independent risk factor for sarcopenia and reduced grip strength.Anemia is associated with sarcopenia,but is not an independent risk factor for sarcopenia.

Aim To explore the role and mechanism of epimedokoreanin B(EKB)in HepaRG cell pyroptosis through endoplasmic reticulum stress and NLRP1-me-diated pyroptosis pathway.Methods The effect of EKB on the viability of HepaRG cells at different con-centrations was determined by MTT assay,and the cell growth status was recorded by Incucyte.Four groups of HepaRG cells were set up.The control group was cul-tured with complete medium for 24 h;the drug admin-istration group was cultured with three concentration gradients of 6.25,12.5 and 25 μmol·L-1 of EKB for 24 h.Western blot was used to detect the expression levels of endoplasmic reticulum stress-related proteins and pyroptosis-related proteins in the cells of each group.Results HepaRG cells showed cytotoxicity at a concentration of 6.25 μmol·L-1 for 24 h,and the half maximal inhibitory concentration(IC50)was 12.41 μmol·L-1.Incucyte recordings of the cell growth status showed that the cells in the control group were in good growth status,and the vesicular pyropto-sis cells appeared in the different concentrations of EKB and the cells swelled and ruptured after 24 h.Western blot showed that the protein expression levels of endoplasmic reticulum stress-related proteins pERK,eIF-2α,ATF-4,GRP78,and CHOP significantly in-creased in HepaRG cells at 25 μmol·L-1 of EKB compared with the control group.The proteins of the classical pathway of cellular pyroptosis mediated by NLRP1,caspase-1,cleaved caspase-1,GSDMD,GS-DMD-N significantly increased in HepaRG cells.Con-clusion EKB administration induces HepaRG cell py-roptosis,and EKB activates HepaRG cells to undergo endoplasmic reticulum stress and activates the NLRP1/caspase-1/GSDMD-mediated pyroptosis pathway.

Aim To study the difference in hepatotox-icity of psoralen on normal rats and Yin-deficiency rats from the perspective of lipid metabolism,so as to help explain the mechanism of psoralen cautiously used in patients with Yin deficiency recorded in ancient books.Methods SD rats were randomly divided into the nor-mal control group(carboxymethyl cellulose-Na,CMC-Na),normal administration group(CMC-Na+psor-alen),Yin-deficiency control group(CMC-Na+thy-roxine)and Yin-deficiency administration group(CMC-Na+thyroxine+psoralen).The model of Yin-deficiency was established by thyroxine(1 mg·kg-1)for ten days,and then psoralen(200 mg·kg-1)was given for three days.The serum indexes related to liver injury were detected by automatic biochemical analy-zer,the morphological changes of liver tissue were ob-served using HE and oil red O staining,and the relative transcription levels of lipid metabolism related enzymes and mRNA of transporter and endoplasmic reticulum stress related factors were detected using Real-time PCR.Results After intragastric administration of psoralen for three days,compared with the normal group,the levels of serum alanine aminotransferase(ALT),aspartate transaminase(AST),total bile acid(TBA)and triglyeride(TG)in Yin deficiency group increased more significantly,while TC,ALB and TP de-creased more significantly,and liver HE and oil red O staining showed more obvious lipid degeneration.TG synthesis factors adrenocortical carcinoma(ACC),fatty acid synthase(FASN)and sterolregulatory element binding protein-1(SREBP-1)were down-regulated more significantly,TG transport factors mili-total pro-tein(MTP)and lipoprotein pipase(LPL)were down-regulated more evidently,fatty acid β-oxidation related factors carnitine palmitoyltransferase 1A(CPT1A),carnitine/organic cation transporter 2(OCTN2)and peroxisome proliferators-activated receptors-alpha(PPARα)were down-regulated more apparently,TC transporter adenosine triphosphate binding cassette transporter G8(ABCG8)and bile acid receptor farne-soid X receptor(FXR)were down-regulated more ob-viously,and endoplasmic reticulum stress factor activa-ting transcription factor 4(ATF4)was up-regulated more significantly.Conclusions Psoralen can cause more severe hepatotoxicity in Yin deficiency rats than that in normal administration group,and its mechanism may be related to the disorder of hepatic lipid metabo-lism,aggravation of hepatic cholestasis and steatosis,and activation of endoplasmic reticulum stress re-sponse.

The phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)signaling pathway plays a crucial role in the regulation of renal fibrosis by participating in inflammatory response,oxidative stress and autophagy.Paeoniflorin exhibits remarkable efficacy in treating myocardial and liver fibrosis.This article provides a comprehensive review on the research progress of paeoniflora in preventing and treating renal fibrosis through modulation of the PI3K/Akt/mTOR signaling pathway,offering novel insights for traditional Chinese medicine-based approaches to prevent and treat renal fibrosis.

Objective:To optimize the previously established Improved Membrane Filtration(ISET)method for de-tecting circulating tumor cells(CTCs)and to develop and validate a new,reliable CTC detection method by combining it with immunofluorescence techniques.Methods:The study involved optimizing the CTC detection system using mixed samples of gastric cancer 803 cell lines and peripheral blood(PB)from healthy volunteers to simulate the peripheral blood of cancer patients.The optimized system was validated using peripheral blood samples from 23 patients with ad-vanced(Ⅲ/Ⅳ)cancer,employing the ISET technique combined with immunocytochemistry(ICC)and paraffin block im-munohistochemistry(IHC)identification techniques.A cohort of 74 patients with various cancer types was used to com-pare the diagnostic performance of the membrane filtration combined with immunofluorescence(ISET-ICC)system against the CTC-Biopsy and CellSearchTM methods.Results:By adding a red blood cell lysis step and increasing the membrane pore size to 10 μm,the filtration time of the ISET method was reduced by threefold.The ISET-ICC detection method achieved a CTC positivity rate of 65.2％.The combination of the ISET-ICC system with the cell paraffin block-IHC method enhanced the reliability of identifying circulating tumor cells undergoing epithelial-mesenchymal transition(EMT-CTCs).The ISET-ICC technique demonstrated a significantly higher CTC positivity rate(17.6％;13/74)within the study cohort compared to the CellSearchTM method,which showed a CTC positivity rate of 12.2％(9/74;x2=10.21,P=0.007).The difference in positive detection rates among patients at different stages was statistically significant(x2=3.64,P=0.029),with a notably higher CTC detection rate in stage Ⅳ patients compared to those in stages Ⅰ-Ⅲ(x2=6.76,P=0.001).Conclusion:The improved ISET-ICC system effectively detects CTCs across various cancer types and dem-onstrates greater accuracy compared to the CellSearchTM system.

The study aimed to investigate the effect of the CD200R1 gene deletion on microglia activation and nigrostriatal dopamine neuron loss in the Parkinson's disease (PD) process. The CRISPR-Cas9 technology was applied to construct the CD200R1^-/- mice. The primary microglia cells of wild-type and CD200R1^-/- mice were cultured and treated with bacterial lipopolysaccharide (LPS). Microglia phagocytosis level was assessed by a fluorescent microsphere phagocytosis assay. PD mouse model was prepared by nigral stereotaxic injection of recombinant adeno-associated virus vector carrying human α-synuclein (α-syn). The changes in the motor behavior of the mice with both genotypes were evaluated by cylinder test, open field test, and rotarod test. Immunohistochemical staining was used to assess the loss of dopamine neurons in substantia nigra. Immunofluorescence staining was used to detect the expression level of CD68 (a key molecule involved in phagocytosis) in microglia. The results showed that CD200R1 deletion markedly enhanced LPS-induced phagocytosis in vitro by the microglial cells. In the mouse model of PD, CD200R1 deletion exacerbated motor behavior impairment and dopamine neuron loss in substantia nigra. Fluorescence intensity analysis results revealed a significant increase in CD68 expression in microglia located in the substantia nigra of CD200R1^-/- mice. The above results suggest that CD200R1 deletion may further activates microglia by promoting microglial phagocytosis, leading to increased loss of the nigrostriatal dopamine neurons in the PD model mice. Therefore, targeting CD200R1 could potentially serve as a novel therapeutic target for the treatment of early-stage PD.
Animals ; Microglia/physiology* ; Mice ; Phagocytosis ; Parkinson Disease/genetics* ; Disease Models, Animal ; Receptors, Cell Surface/physiology* ; Dopaminergic Neurons/pathology* ; Antigens, CD/metabolism* ; Gene Deletion ; Substantia Nigra ; Mice, Inbred C57BL ; Mice, Knockout ; Cells, Cultured ; Male ; alpha-Synuclein ; CD68 Molecule ; Orexin Receptors