Objective To investigate the factors influencing the natural outcome of low-grade squamous intraepithelial lesions(LSIL)of the cervix.Methods A total of 154 patients who underwent colposcopic cervical biopsy in the Third Affiliated Hospital of Zhengzhou University from January 2022 to January 2023 were selected and divided into negative conversion group(55 cases),continuous group(70 cases)and upgraded group(29 cases)according to the follow-up results.Logistic regression was used to analyze the influence of related factors on the outcome of LSIL.Results There were statistically significant differences in the number of vaginal births,the results of thin-prep cytology test,human papilloma virus(HPV)typing,whether vaginal LSIL was combined,whether there were symptoms,and vaginal microecology among three groups(P＜0.05).Multivariate Logistic analysis showed that combined vaginal LSIL,vaginal microecological abnormalities,atypical squamous cell-cannot exclude high-grade squamous intraepithelial lesion(ASC-H)/high-grade squamous intraepithelial lesion(HSIL),HPV 16/18 positive or mixed positive were independent risk factors for persistent infection(P＜0.05),combined vaginal LSIL,symptomatic,vaginal microecological abnormalities,ASC-H/HSIL,HPV 16/18 positive or mixed positive were all independent risk factors for escalation of lesions(P＜0.05).Conclusion Patients with vaginal LSIL,vaginal microecological abnormalities,ASC-H/HSIL,HPV 16/18 positive or mixed positive should be followed up and early intervention to reduce the risk of disease escalation.

Objective:To explore the phenotypic heterogeneity among patients harboring identical pathogenic variants in the dystrophin ( DMD) gene by analyzing clinical and imaging data from 7 pairs of male twins with dystrophinopathy. Methods:Clinical and laboratory data of 14 (7 pairs) male twins diagnosed with dystrophinopathy through genetic testing among 1 767 patients at Peking University First Hospital from January 2017 to October 2024 were collected. Eleven patients underwent thigh muscle magnetic resonance imaging (MRI), and muscle biopsies were performed in at least 1 case of each pair.Results:Among the 7 pairs of twin patients, 2 pairs had Duchenne muscular dystrophy, and 5 pairs had Becker muscular dystrophy. In terms of variant types, 4 pairs had in-frame deletions, while the remaining 3 pairs had duplication variants, frameshift variants, and nonsense variants, respectively. Clinically, 6 individuals had asymptomatic hypercreatine kinasemia, and 8 had varying degrees of limb weakness. Among the 5 pairs of symptomatic twins, there were differences in the degree of limb weakness. Four individuals showed no significant abnormalities in thigh muscle MRI, 7 showed fat infiltration mainly in the bilateral gluteus maximus and adductor magnus muscles, and 2 pairs of twins had obvious differences in the degree of fat infiltration in muscle MRI. Muscle biopsies revealed dystrophic or mild myopathic pathological changes, with 2 individuals showing severe loss of dystrophin, while the others had partial loss.Conclusions:Dystrophinopathy exhibits significant individual differences. Even among individuals with highly similar genetic background, clinical and imaging manifestations caused by the same pathogenic variant also vary.

Objective To explore the effect of chrysin on intestinal flora in mice with alcoholic liver disease(ALD).Methods Mice were randomly assigned to normal control group,ALD model group,Silymarin group,chrysin low-dose group,chrysin high-dose group(25,50 mg·kg-1).The mice were fed with alcoholic liquid diet and a single dose of alcohol(5 g·kg-1)for eight weeks to establish the ALD model.After eight weeks of oral administration,each group's serum and plasma lipids and liver function indices were collected and detected using kits;then collected the liver and observed the pathological changes of the liver using HE staining;meanwhile,intestinal contents were collected and changes in mouse gut flora were analyzed by 16S rDNA sequencing.Results Compared with the ALD group,the level of aspartate transaminase(AST),alanine transaminase(ALT)and triacylglycerol(TG)of low-dose and high-dose chrysin groups were significantly reduced,and it can alleviate liver cell steatosis and inflammatory reactions caused by alcohol.16S rDNA results showed that the total number and types of intestinal flora in the ethanol group were significantly reduced,as well as a change in the dominant genus to Escherichia-Shigella and Akkermansia.Compared to the ALD model group,the Shannon index of the intestinal microbiota increased significantly in mice treated with low and high doses of chrysin.In addition,at the phylum and genus level,the abundance of the high-dose chrysin group increased significantly,resulting in an overall increase in the total number and amount of microbiota.The abundance of dominant bacterial groups,such as Oscillospirales,irmicutes,andAlloprevotella,was also significantly increased.Conclusion Chrysin may exert therapeutic effects on ALD by improving intestinal flora imbalance in ALD mice.

Objective To investigate the factors influencing the natural outcome of low-grade squamous intraepithelial lesions(LSIL)of the cervix.Methods A total of 154 patients who underwent colposcopic cervical biopsy in the Third Affiliated Hospital of Zhengzhou University from January 2022 to January 2023 were selected and divided into negative conversion group(55 cases),continuous group(70 cases)and upgraded group(29 cases)according to the follow-up results.Logistic regression was used to analyze the influence of related factors on the outcome of LSIL.Results There were statistically significant differences in the number of vaginal births,the results of thin-prep cytology test,human papilloma virus(HPV)typing,whether vaginal LSIL was combined,whether there were symptoms,and vaginal microecology among three groups(P＜0.05).Multivariate Logistic analysis showed that combined vaginal LSIL,vaginal microecological abnormalities,atypical squamous cell-cannot exclude high-grade squamous intraepithelial lesion(ASC-H)/high-grade squamous intraepithelial lesion(HSIL),HPV 16/18 positive or mixed positive were independent risk factors for persistent infection(P＜0.05),combined vaginal LSIL,symptomatic,vaginal microecological abnormalities,ASC-H/HSIL,HPV 16/18 positive or mixed positive were all independent risk factors for escalation of lesions(P＜0.05).Conclusion Patients with vaginal LSIL,vaginal microecological abnormalities,ASC-H/HSIL,HPV 16/18 positive or mixed positive should be followed up and early intervention to reduce the risk of disease escalation.

Objective To explore the effect of chrysin on intestinal flora in mice with alcoholic liver disease(ALD).Methods Mice were randomly assigned to normal control group,ALD model group,Silymarin group,chrysin low-dose group,chrysin high-dose group(25,50 mg·kg-1).The mice were fed with alcoholic liquid diet and a single dose of alcohol(5 g·kg-1)for eight weeks to establish the ALD model.After eight weeks of oral administration,each group's serum and plasma lipids and liver function indices were collected and detected using kits;then collected the liver and observed the pathological changes of the liver using HE staining;meanwhile,intestinal contents were collected and changes in mouse gut flora were analyzed by 16S rDNA sequencing.Results Compared with the ALD group,the level of aspartate transaminase(AST),alanine transaminase(ALT)and triacylglycerol(TG)of low-dose and high-dose chrysin groups were significantly reduced,and it can alleviate liver cell steatosis and inflammatory reactions caused by alcohol.16S rDNA results showed that the total number and types of intestinal flora in the ethanol group were significantly reduced,as well as a change in the dominant genus to Escherichia-Shigella and Akkermansia.Compared to the ALD model group,the Shannon index of the intestinal microbiota increased significantly in mice treated with low and high doses of chrysin.In addition,at the phylum and genus level,the abundance of the high-dose chrysin group increased significantly,resulting in an overall increase in the total number and amount of microbiota.The abundance of dominant bacterial groups,such as Oscillospirales,irmicutes,andAlloprevotella,was also significantly increased.Conclusion Chrysin may exert therapeutic effects on ALD by improving intestinal flora imbalance in ALD mice.

Objective:To explore the phenotypic heterogeneity among patients harboring identical pathogenic variants in the dystrophin ( DMD) gene by analyzing clinical and imaging data from 7 pairs of male twins with dystrophinopathy. Methods:Clinical and laboratory data of 14 (7 pairs) male twins diagnosed with dystrophinopathy through genetic testing among 1 767 patients at Peking University First Hospital from January 2017 to October 2024 were collected. Eleven patients underwent thigh muscle magnetic resonance imaging (MRI), and muscle biopsies were performed in at least 1 case of each pair.Results:Among the 7 pairs of twin patients, 2 pairs had Duchenne muscular dystrophy, and 5 pairs had Becker muscular dystrophy. In terms of variant types, 4 pairs had in-frame deletions, while the remaining 3 pairs had duplication variants, frameshift variants, and nonsense variants, respectively. Clinically, 6 individuals had asymptomatic hypercreatine kinasemia, and 8 had varying degrees of limb weakness. Among the 5 pairs of symptomatic twins, there were differences in the degree of limb weakness. Four individuals showed no significant abnormalities in thigh muscle MRI, 7 showed fat infiltration mainly in the bilateral gluteus maximus and adductor magnus muscles, and 2 pairs of twins had obvious differences in the degree of fat infiltration in muscle MRI. Muscle biopsies revealed dystrophic or mild myopathic pathological changes, with 2 individuals showing severe loss of dystrophin, while the others had partial loss.Conclusions:Dystrophinopathy exhibits significant individual differences. Even among individuals with highly similar genetic background, clinical and imaging manifestations caused by the same pathogenic variant also vary.

Objective:To investigate the effect of decarbromodiphenyl ether(BDE-209)exposure on the migration ability of triple-negative breast cancer(TNBC)cells and to explore the underlying mechanism.Methods:Human TNBC MDA-MB-231 cells were divided into blank control group and BDE-209 exposure groups(treated with 0.02,0.20,2.00,20.00 and 200.00 ng/mL BDE-209 in high glucose DMEM).Extracellular vehicles(EVs)secreted by MDA-MB-231 cells were isolated by differential ultracentrifugation.Transmission electron microscopy(SEM),nanoparticle tracking analysis(NTA)and Western blotting were performed to characterize the EVs.The effect of the EVs induced by BDE-209 exposure(EVs-BDE-209)on the migration and invasion of MDA-MB-231 cells was detected by wound-healing assay and Transwell test.qRT-PCR was used to measure the miR-221 level in EVs-BDE-209.The expression of MMP9 in MDA-MB-231 cells was determined by Western blotting.Results:Compared with the blank control,BDE-209 exposure increased the tumor cell-derived EVs in dose-dependent manner.The MDA-MB-231 cells co-cultured with EVs released by 200.00 ng/mL BDE-209 exposure showed an 86%increase in cell migration rate,a 1.32-fold higher number of membrane-penetrating cells,a 2.71-fold higher expression level of miR-221,and a 1.62-fold higher expression level of MMP9 compared with the blank control group(all P<0.05).While transfection with anti-miR-221 antibody to decrease miR-221 level in EVs significantly reversed the increased invasion ability of the MDA-MB-231 cells treated with EVs-BDE-209.Conclusion:BDE-209 exposure may promote metastasis potential of MDA-MB-231 cells via EVs-BDE-209 transmitted miR-221.

Objective Mitochondrial reactive oxygen species(mtROS)could cause damage to pancreatic β-cells,rendering them susceptible to oxidative damage.Hence,investigating the potential of the mitochondria-targeted antioxidant(Mito-TEMPO)to protect pancreatic β-cells from ferroptosis by mitigating lipid peroxidation becomes crucial. Methods MIN6 cells were cultured in vitro with 100 μmol/L sodium palmitate(SP)to simulate diabetes.FerroOrange was utilized for the detection of Fe2+fluorescence staining,BODIPY581/591C11 for lipid reactive oxygen species,and MitoSox-Red for mtROS.Alterations in mitophagy levels were assessed through the co-localization of lysosomal and mitochondrial fluorescence.Western blotting was employed to quantify protein levels of Acsl4,GPX4,FSP1,FE,PINK1,Parkin,TOMM20,P62,and LC3.Subsequently,interventions were implemented using Mito-TEMPO and Carbonyl cyanide 3-chlorophenylhydrazone(CCCP)to observe changes in ferroptosis and mitophagy within MIN6 cells. Results We found that SP induced a dose-dependent increase in Fe2+and lipid ROS in MIN6 cells while decreasing the expression levels of GPX4 and FSP1 proteins.Through bioinformatics analysis,it has been uncovered that mitophagy assumes a crucial role within the ferroptosis pathway associated with diabetes.Additionally,SP decreased the expression of mitophagy-related proteins PINK1 and Parkin,leading to mtROS overproduction.Conversely,Mito-TEMPO effectively eliminated mtROS while activating the mitophagy pathways involving PINK1 and Parkin,thereby reducing the occurrence of ferroptosis in MIN6 cells.CCCP also demonstrated efficacy in reducing ferroptosis in MIN6 cells. Conclusion In summary,Mito-TEMPO proved effective in attenuating mtROS production and initiating mitophagy pathways mediated by PINK1 and Parkin in MIN6 cells.Consequently,this decreased iron overload and lipid peroxidation,ultimately safeguarding the cells from ferroptosis.

Objective:To evaluate the effect of gastrogin on AMP-activated protein kinase (AMPK)/transient receptor potential anchor protein 1 (TRPA1) signaling pathway in rats with neuropathic pain.Methods:Thirty-six SPF-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-230 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation+ normal saline group (SHAM group), neuropathic pain+ normal saline group (NP group), and neuropathic pain+ gastrogin group (GAS group). Neuropathic pain was induced by chronic constrictive injury to sciatic nerve under 2% isoflurane anaesthesia. The sciatic nerve was only exposed but not ligated in SHAM group. Gastrogin 100 mg/kg was intraperitoneally injected for 14 consecutive days after developing the model in GAS group, while the equal volume of normal saline was given instead in SHAM and NP groups. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before developing the model (T 0) and 1, 3, 5, 7, 10 and 14 days after developing the model (T 1-6). The rats were anesthetized and sacrificed following the measurement of pain thresholds at T 4 and T 6. The lumbar segment (L 4-6) of the spinal cord was removed for determination of TRPA1 mRNA expression (by quantitative real-time polymerase chain reaction), expression of TRPA1, AMPK and p-AMPK (by Western blot), expression of TRPA1 (by immunofluorescence staining) and expression of tumor necrosis-alpha(TNF-α), interleukin-1beta(IL-1β) and c-fos (by immunohistochemistry). Results:Compared with SHAM group, MWT and TWL were significantly decreased at T 1-6, the expression of TRPA1 mRNA, TRPA1, TNF-α, IL-1β and c-fos was up-regulated, the expression of p-AMPK was down-regulated ( P<0.05), and no significant change was found in AMPK expression in NP group ( P>0.05). Compared with NP group, MWT at T 3-6 and TWL at T 2-6 were significantly increased, the expression of TRPA1 mRNA, TRPA1, TNF-α, IL-1β and c-fos was down-regulated, and p-AMPK expression was up-regulated ( P<0.05), and no significant change was found in AMPK expression in GAS group ( P>0.05). Conclusions:The mechanism by which gastrogin reduces neuropathic pain may be related to modulating the expression of the AMPK/TRPA1 signaling pathway in rats.

OBJECTIVE To study the effects of bergapten in the treatment of liver fibrosis and its mechanism based on serum metabolomics. METHODS Forty mice were divided into normal control group （0.5% carboxymethyl cellulose sodium solution）， model group （0.5% carboxymethyl cellulose sodium solution）， and BP low-dose and high-dose groups （50， 100 mg/kg）， with 10 mice in each group. Except for the normal control group， the other three groups were all treated with carbon tetrachloride to induce liver fibrosis model； they were given relevant medicine/solution intragastrically， once a day， for consecutive 8 weeks. After the last medication， the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum were detected， and liver pathological changes were observed； the expressions of α-smooth muscle actin （α-SMA） and Collagen Ⅰ were detected in liver tissue； the serum of the mice was collected for metabolomics analysis. RESULTS Compared with the model group， serum levels of ALT and AST and protein expressions of α-SMA and Collagen Ⅰ in liver tissue were decreased significantly in BP high-dose and low-dose groups （P＜0.05）， while liver fibrosis was improved significantly. Meanwhile， metabolomics analyses showed that there were a total of 175 serum differential metabolites in the BP high-dose group and model group， of which 18 substances were upregulated and 157 substances were downregulated； the main metabolic pathways involved in bergapten intervention were pyrimidine metabolism， butanoate metabolism， fatty acid synthesis， tyrosine metabolism， β-alanine metabolism， nicotinic acid and nicotinamide metabolism， glutathione metabolism， etc. CONCLUSIONS BP is effective in the treatment of liver fibrosis by regulating pyrimidine metabolism， butanoate metabolism， glutathione metabolism and so on in rats with liver fibrosis.