In this report, the protective effect and molecular mechanism of Chaihu Shugan San-containing serum on corticosterone(CORT)-induced mitochondrial damage in pheochromocytoma(PC12) cells was studied based on CORT-induced rat PC12 cell model. The cultured cells were divided into five groups: blank control group, CORT group(400 μmol·L~(-1) CORT), Chaihu Shugan San-containing serum group(400 μmol·L~(-1) CORT + 10% Chaihu Shugan San-containing serum), control serum group(400 μmol·L~(-1) CORT + 10% control serum), and fluoxetine group(400 μmol·L~(-1) CORT + 10% fluoxetine-containing serum). The study was carried out by cell activity detection, mitochondrial morphology observation, membrane potential measurement, energy metabolism analysis, and mitochondrial dynamics-related protein detection. The results showed that CORT treatment significantly reduced the survival rate of PC12 cells, altered mitochondrial morphology, and decreased mitochondrial membrane potential and adenosine triphosphate(ATP) synthetic rate. Both Chaihu Shugan San-and fluoxetine-containing serum significantly increased the survival rate of CORT-treated PC12 cells and the ATP synthetic rate in the mitochondria. Unlike fluoxetine, Chaihu Shugan San-containing serum significantly inhibited the decrease in mitochondrial membrane potential caused by CORT and increased the oxygen consumption rate(OCR) values of both mitochondrial maximum respiration and reserve respiration capacity. Western blot analysis showed that CORT induced upregulated protein expressions of dynamin-related protein 1(Drp1) and peroxisome proliferator-activated receptor gamma co-activator 1α(PGC-1α) in PC12 cells and specific protein expression of optic atrophy protein 1(OPA1), yet it repressed the protein expressions of silent information regulator 1(SIRT1) and mitochondrial fusion protein 1(Mfn1) in PC12 cells. Both Chaihu Shugan San-and fluoxetine-containing serum significantly inhibited the protein expression of Drp1. However, only Chaihu Shugan San-containing serum could significantly inhibit the CORT-induced upregulation protein of PGC-1α. RESULTS:: herein suggest that Chaihu Shugan San-containing serum can alleviate CORT-induced damage in PC12 cells, which may be related to the mitochondrial fragmentation/lipid peroxidation protection by Drp1 inhibition, as well as mitochondrial dynamics and energy metabolism mediated by PGC-1α/SIRT1 signaling pathway.
Animals ; PC12 Cells ; Rats ; Mitochondrial Dynamics/drug effects* ; Mitochondria/metabolism* ; Corticosterone/adverse effects* ; Membrane Potential, Mitochondrial/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Protective Agents/pharmacology* ; Cell Survival/drug effects*

Solid-phase extraction(SPE)combined with surface-enhanced Raman spectroscopy(SERS)has emerged as a promising analytical technique for detection and analysis of trace components in complex sample matrices.SPE enriches analytes through selective adsorption and solvent elution,effectively increasing the concentration and signal intensity.SERS enables ultra-sensitive and highly selective molecular analysis through the use of SERS-active substrates engineered to amplify Raman signals.The integration of these two techniques overcomes the limitations of conventional Raman spectroscopy in low-concentration detection field,while significantly improving sample preparation efficiency and analytical accuracy.This review provided a comprehensive overview of the characteristics of three SPE-SERS coupling modes,including two-step,one-step,and online integration.Special emphasis was placed on recent advancements in one-step SPE-SERS approaches based on novel functional adsorbent materials such as graphene,metal-organic frameworks,covalent organic frameworks,and molecularly imprinted polymers.Furthermore,future directions and development prospects of SPE-SERS technology were discussed.

AIM To investigate the effects of Huazhuo Jiedu Shugan Formula(HJSGF)on improving learning and memory impairment in a rat model of vascular dementia(VD)via SIRT1/PGC-1α/PPARγ pathway.METHODS The SD rats were randomly divided into the sham control group,the model group,the donepezil group(0.5 mg/kg),and the low-,medium-and high-dose HJSGF groups(2.7,5.4,10.8 g/kg),with 10 rats in each group.The VD rat models established by bilateral common carotid artery permanent ligation(2-VO)had their neurological behavior assessment using the Longa5-point scale,and their modeling success confirmed by the Morris water maze test and their 3-week corresponding dosing of drugs by gavage afterward.After the drug administration,the rats had their spatial memory ability tested through behavioral experiments;their serum levels of IL-18 and IL-1β measured by ELISA;their histopathological changes and neuronal morphology in the hippocampal CA1 region observed by HE staining and Nissl staining;and their hippocampal protein expressions of SIRT1,PGC-1α and PPARγ detected by immunohistochemistry and Western blot.RESULTS Compared with the sham control group,the model group showed prolonged escape latency(P＜0.01);decreased platform crossing times and target quadrant residence time(P＜0.01);disorganized arrangement of hippocampal CA1 neurons,nuclear condensation,reduced Nissl bodies,increased secretion and protein expressions of IL-1β and IL-18(P＜0.01);and reduced hippocampal protein expressions of SIRT1,PGC-1α and PPARγ(P＜0.01).Compared with the model group,the groups intervened with donepezil or HJSGF showed shortened escape latency(P＜0.05,P＜0.01);increased platform crossing times and target quadrant residence time(P＜0.05,P＜0.01);alleviated damage of the hippocampal CA1 region,reduced secretion and protein expressions of IL-1β and IL-18(P＜0.05,P＜0.01);and elevated hippocampal protein expressions of SIRT1,PGC-1α and PPARγ(P＜0.05,P＜0.01).CONCLUSION HJSGF may alleviate the inflammatory responses in VD rats and therefore improve their learning and memory impairment by activating the SIRT1/PGC-1α/PPARγ signaling pathway.

Objective The objective of this study was to explore the effect of knocking down CXCL8 on the efficacy of pacli-taxel chemotherapy in cervical squamous cell carcinoma and to investigate its potential mechanism of action.Methods The Hela cell model was used to specifically inhibit CXCL8 gene expression through lentivirus-mediated RNA interference(RNAi)technology.The optimal transfection conditions were HitransG P and a multiplicity of infection(MOI)of 100.The CCK-8 assay was used to screen the optimal intervention concentration of puromycin as 1.5μg/mL.The LV-CXCL8-RNAi(3 targets)and negative control lentivirus were transfected into cells under optimal transfection conditions and set up a blank control group.The qRT-PCR assay was used to select the sh-CXCL8-13 group lentivirus as the intervention sequence and virus for subsequent experiments.The experiments were divided into the blank control group,negative control group,sh-CXCL8 group,paclitaxel group,and sh-CXCL8+paclitaxel group.The prolifer-ative activity and invasive ability of cervical cancer cells were assessed by CCK-8 and cell invasion assays.The expression of CXCL8,Bcl2,Bax,and β-actin were detected by qRT-PCR and Western blot.Results Compared with the other four groups,the proliferative and invasive ability of Hela cells was significantly reduced in the sh-CXCL8+paclitaxel group,and the difference was statistically sig-nificant(P<0.01).The qRT-PCR results showed that the expression of CXCL8,Bcl2,PIK3CB,and Akt1 genes was significantly re-duced,and the expression of Bax gene was significantly increased in the sh-CXCL8+paclitaxel group.The difference between the groups was statistically significant(P<0.001).The results of Western blot showed that the expression of CXCL8,PIK3CB,and p-Akt1 proteins was reduced in the sh-CXCL8+paclitaxel group(P<0.05).Conclusion Knocking down CXCL8 can reduce the prolif-erative and invasive capacity of Hela cells,possibly by affecting the PI3K/Akt pathway to affect the drug sensitivity of Hela cells to paclitaxel.

Objective To assess the expression of CXCL8 in cervical cancer and its association with clinicopathological features and therapeutic efficacy of patients.Methods Bioinformatic analysis was performed using The Cancer Genome Atlas and Genotype-Tissue Expression databases to compare CXCL8 expression between cervical cancer and normal tissues.R software(version 4.4.0)was used for data analysis,with the timeROC package applied to construct time-dependent receiver operating characteristic(ROC)curves for evalua-ting the prognostic predictive efficacy of CXCL8.The survival package with the survfit function was used to compare survival differences between CXCL8 high-and low-expression groups.Clinical data and tissue specimens were collected from 94 patients with cervical squa-mous cell cancer treated at The First Affiliated Hospital of Xinjiang Medical University between January 2017 and December 2021.Immu-nohistochemical staining was used to detect CXCL8 expression levels and analyze its correlation with clinicopathological characteristics,therapeutic efficacy,and prognosis.Results Bioinformatic analysis showed that CXCL8 was highly expressed in cervical cancer tissues than in normal tissues(P＜0.05).Time-dependent ROC curves and survival analyses showed that patients with high CXCL8 expression had significantly shorter overall survival than those with low CXCL8 expression(P＜0.001).Immunohistochemical results showed that CXCL8 expression in cervical cancer tissues was significantly higher than that in adjacent tissues(P＜0.000 1).Clinical correlation analy-sis revealed that CXCL8 expression levels were associated with treatment regimen(P＜0.001)and short-term therapeutic efficacy(P=0.017).Compared to the low-expression group,the high-expression group showed a significantly lower therapeutic efficacy and shorter overall survival(P＜0.05).Conclusion CXCL8 is highly expressed in cervical cancer tissues,and patients with high CXCL8 expression have poor prognosis.Thus,CXCL8 may be an effective target for assessing the prognosis and clinical treatment of cervical cancer.

Objective To assess the expression of CXCL8 in cervical cancer and its association with clinicopathological features and therapeutic efficacy of patients.Methods Bioinformatic analysis was performed using The Cancer Genome Atlas and Genotype-Tissue Expression databases to compare CXCL8 expression between cervical cancer and normal tissues.R software(version 4.4.0)was used for data analysis,with the timeROC package applied to construct time-dependent receiver operating characteristic(ROC)curves for evalua-ting the prognostic predictive efficacy of CXCL8.The survival package with the survfit function was used to compare survival differences between CXCL8 high-and low-expression groups.Clinical data and tissue specimens were collected from 94 patients with cervical squa-mous cell cancer treated at The First Affiliated Hospital of Xinjiang Medical University between January 2017 and December 2021.Immu-nohistochemical staining was used to detect CXCL8 expression levels and analyze its correlation with clinicopathological characteristics,therapeutic efficacy,and prognosis.Results Bioinformatic analysis showed that CXCL8 was highly expressed in cervical cancer tissues than in normal tissues(P＜0.05).Time-dependent ROC curves and survival analyses showed that patients with high CXCL8 expression had significantly shorter overall survival than those with low CXCL8 expression(P＜0.001).Immunohistochemical results showed that CXCL8 expression in cervical cancer tissues was significantly higher than that in adjacent tissues(P＜0.000 1).Clinical correlation analy-sis revealed that CXCL8 expression levels were associated with treatment regimen(P＜0.001)and short-term therapeutic efficacy(P=0.017).Compared to the low-expression group,the high-expression group showed a significantly lower therapeutic efficacy and shorter overall survival(P＜0.05).Conclusion CXCL8 is highly expressed in cervical cancer tissues,and patients with high CXCL8 expression have poor prognosis.Thus,CXCL8 may be an effective target for assessing the prognosis and clinical treatment of cervical cancer.

AIM To investigate the effects of Huazhuo Jiedu Shugan Formula(HJSGF)on improving learning and memory impairment in a rat model of vascular dementia(VD)via SIRT1/PGC-1α/PPARγ pathway.METHODS The SD rats were randomly divided into the sham control group,the model group,the donepezil group(0.5 mg/kg),and the low-,medium-and high-dose HJSGF groups(2.7,5.4,10.8 g/kg),with 10 rats in each group.The VD rat models established by bilateral common carotid artery permanent ligation(2-VO)had their neurological behavior assessment using the Longa5-point scale,and their modeling success confirmed by the Morris water maze test and their 3-week corresponding dosing of drugs by gavage afterward.After the drug administration,the rats had their spatial memory ability tested through behavioral experiments;their serum levels of IL-18 and IL-1β measured by ELISA;their histopathological changes and neuronal morphology in the hippocampal CA1 region observed by HE staining and Nissl staining;and their hippocampal protein expressions of SIRT1,PGC-1α and PPARγ detected by immunohistochemistry and Western blot.RESULTS Compared with the sham control group,the model group showed prolonged escape latency(P＜0.01);decreased platform crossing times and target quadrant residence time(P＜0.01);disorganized arrangement of hippocampal CA1 neurons,nuclear condensation,reduced Nissl bodies,increased secretion and protein expressions of IL-1β and IL-18(P＜0.01);and reduced hippocampal protein expressions of SIRT1,PGC-1α and PPARγ(P＜0.01).Compared with the model group,the groups intervened with donepezil or HJSGF showed shortened escape latency(P＜0.05,P＜0.01);increased platform crossing times and target quadrant residence time(P＜0.05,P＜0.01);alleviated damage of the hippocampal CA1 region,reduced secretion and protein expressions of IL-1β and IL-18(P＜0.05,P＜0.01);and elevated hippocampal protein expressions of SIRT1,PGC-1α and PPARγ(P＜0.05,P＜0.01).CONCLUSION HJSGF may alleviate the inflammatory responses in VD rats and therefore improve their learning and memory impairment by activating the SIRT1/PGC-1α/PPARγ signaling pathway.

Objective The objective of this study was to explore the effect of knocking down CXCL8 on the efficacy of pacli-taxel chemotherapy in cervical squamous cell carcinoma and to investigate its potential mechanism of action.Methods The Hela cell model was used to specifically inhibit CXCL8 gene expression through lentivirus-mediated RNA interference(RNAi)technology.The optimal transfection conditions were HitransG P and a multiplicity of infection(MOI)of 100.The CCK-8 assay was used to screen the optimal intervention concentration of puromycin as 1.5μg/mL.The LV-CXCL8-RNAi(3 targets)and negative control lentivirus were transfected into cells under optimal transfection conditions and set up a blank control group.The qRT-PCR assay was used to select the sh-CXCL8-13 group lentivirus as the intervention sequence and virus for subsequent experiments.The experiments were divided into the blank control group,negative control group,sh-CXCL8 group,paclitaxel group,and sh-CXCL8+paclitaxel group.The prolifer-ative activity and invasive ability of cervical cancer cells were assessed by CCK-8 and cell invasion assays.The expression of CXCL8,Bcl2,Bax,and β-actin were detected by qRT-PCR and Western blot.Results Compared with the other four groups,the proliferative and invasive ability of Hela cells was significantly reduced in the sh-CXCL8+paclitaxel group,and the difference was statistically sig-nificant(P<0.01).The qRT-PCR results showed that the expression of CXCL8,Bcl2,PIK3CB,and Akt1 genes was significantly re-duced,and the expression of Bax gene was significantly increased in the sh-CXCL8+paclitaxel group.The difference between the groups was statistically significant(P<0.001).The results of Western blot showed that the expression of CXCL8,PIK3CB,and p-Akt1 proteins was reduced in the sh-CXCL8+paclitaxel group(P<0.05).Conclusion Knocking down CXCL8 can reduce the prolif-erative and invasive capacity of Hela cells,possibly by affecting the PI3K/Akt pathway to affect the drug sensitivity of Hela cells to paclitaxel.

Background/Aims: Despite the high efficacy of direct-acting antivirals (DAAs), approximately 1–3% of hepatitis C virus (HCV) patients fail to achieve a sustained virological response. We conducted a nationwide study to investigate risk factors associated with DAA treatment failure. Machine-learning algorithms have been applied to discriminate subjects who may fail to respond to DAA therapy. Methods: We analyzed the Taiwan HCV Registry Program database to explore predictors of DAA failure in HCV patients. Fifty-five host and virological features were assessed using multivariate logistic regression, decision tree, random forest, eXtreme Gradient Boosting (XGBoost), and artificial neural network. The primary outcome was undetectable HCV RNA at 12 weeks after the end of treatment. Results: The training (n=23,955) and validation (n=10,346) datasets had similar baseline demographics, with an overall DAA failure rate of 1.6% (n=538). Multivariate logistic regression analysis revealed that liver cirrhosis, hepatocellular carcinoma, poor DAA adherence, and higher hemoglobin A1c were significantly associated with virological failure. XGBoost outperformed the other algorithms and logistic regression models, with an area under the receiver operating characteristic curve of 1.000 in the training dataset and 0.803 in the validation dataset. The top five predictors of treatment failure were HCV RNA, body mass index, α-fetoprotein, platelets, and FIB-4 index. The accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of the XGBoost model (cutoff value=0.5) were 99.5%, 69.7%, 99.9%, 97.4%, and 99.5%, respectively, for the entire dataset. Conclusions Machine learning algorithms effectively provide risk stratification for DAA failure and additional information on the factors associated with DAA failure.

The assessment of adverse drug events is an important basis for clinical safety evaluation and post-marketing risk control of drugs,and its causality assessment is gaining increasing attention.The existing methods for assessing the causal relationship between drugs and the occurrence of adverse reactions can be broadly classified into three categories:global introspective methods,standardized methods,and probabilistic methods.At present,there is no systematic introduction of the operational details of the various methods in the domestic literature.This paper compares representative causality assessment methods in terms of definition and concept,methodological steps,industry evaluation and advantages and disadvantages,clarifies the basic process of determining the causality of adverse drug reactions,and discusses how to further improve the adverse drug reaction monitoring and evaluation system,with a view to providing a reference for drug development and pharmacovigilance work in China.