Background/Aims: Cholangiocarcinoma (CCA) is a primary malignant neoplasm with an extremely poor prognosis. While combined chemoradiotherapy has been demonstrated to delay CCA progression to a certain extent, the absence of specific molecular biomarkers or targets significantly hinders the diagnosis and treatment of CCA. Methods: Through cross-analysis of proteomics and ADMA modificationomics, we identified DDX1 overexpressed in CCA with elevated R602-ADMA modifications. HPLC-MS/MS identified PRMT1 as the methyltransferase and USP10 as the deubiquitinating enzyme for DDX1. Immunofluorescence and nuclear-cytoplasmic partitioning experiments confirmed DDX1’s nuclear localization. GO and KEGG analyses clarify the biological functions of DDX1 in response to hypoxia. RNA-seq transcriptomics analyzed key pathways influenced by DDX1. A hydrodynamic in situ CCA mouse model was established to validate the chemopreventive effects of the PRMT1-specific inhibitor GSK715 on CCA development. Results: DDX1 promotes CCA progression both in vivo and in vitro and can be inhibited by GSK715. Mechanistically, PRMT1 mediates ADMA modification at position R602 of DDX1. This modification promotes DDX1 nuclear localization by recruiting USP10 to deubiquitinate DDX1, while simultaneously inhibiting PRMT1 degradation. DDX1 promotes the transcription of PRMT1 and USP10 by binding to the mRNA 3’UTR region, establishing a positive feedback regulatory pathway. This mechanism promotes the occurrence and development of CCA and can serve as a target for the inhibitor GSK715 to suppress CCA progression. Conclusions Our study identified DDX1-R602-ADMA modification as a novel ADMA modification in CCA. It further confirmed its pivotal role in CCA progression. Targeting the USP10-PRMT1-DDX1 axis may represent a significant therapeutic approach for CCA.

Objective To explore the effects of nail-tail transverse connection in the treatment of atlantoaxial dislocation(AAD)and its impacts on bone metabolism,serum vascular endothelial growth factor(VEGF),and fibroblast growth factor-2(FGF-2)levels.Methods A total of 150 pa-tients with AAD were selected as the research subjects and divided into two groups using the random number table method,with 75 cases in each group.The observation group was treated with nail-tail transverse connection combined with posterior atlantoaxial pedicle screw internal fixation(C1-C2 PSR),while the control group was treated with C1-C2 PSR alone.Serum bone metabolism indicators[osteocalcin(BGP),type Ⅰ collagen N-terminal peptide(NTX),bone alkaline phosphatase(BALP),tartrate-resistant acid phosphatase(TRAP),type Ⅰ collagen carboxy-terminal peptide(CTX)],VEGF and FGF-2 levels were compared between the two groups at different time points.The Japanese Orthopaedic Association(JOA)score was used to evaluate the patients' neurological function before surgery and 3 years after surgery.Bone graft fusion was evaluated at 6 months,1 year,2 years,and 3 years after surgery.Results At 1,3 and 6 months after surgery,the serum VEGF and FGF-2 levels in the observation group were higher than those in the control group,and the differences were statistically significant(P＜0.05).After treatment,the BALP,BGP,NTX,TRAP and CTX levels in both groups were lower than those before treatment,and their levels in the observation group were lower than those in the control group,with statistically significant differences(P＜0.05).Before surgery,there was no statistically significant difference in the JOA scores be-tween the two groups(P＞0.05).At 3 years after surgery,the JOA scores in both groups were higher than those before surgery,and the JOA score and the score improvement rate in the observa-tion group were higher than those in the control group,with statistically significant differences(P＜0.05).At 6 months,1 year,2 years and 3 years after surgery,the success rate of bone graft fusion in the observation group was higher than that in the control group,with statistically significant differ-ences(P＜0.05).Conclusion Nail-tail transverse connection has significant effects in the treat-ment of AAD,which can effectively improve patients' bone metabolism and increase the serum VEGF and FGF-2 levels.

Objective:To investigate the protective effects of resveratrol (RSV) on human sperm cryopreservation and explore its underlying protective mechanisms.Methods:A total of 165 normal fresh semen samples were collected from the Reproductive Medicine Center, Department of Obstetrics and Gynecology and Human Sperm Bank of the First Affiliated Hospital of Anhui Medical University between December 2022 and December 2024. Among them, 65 samples were used to obtain semen parameters before and after conventional freezing. Each sample of the other 104 samples was mixed at a 2∶1 volume ratio with cryoprotectant containing 0, 10 -?, 10 -?, or 10 -? mol/L RSV, followed by cryopreservation in liquid nitrogen for 24 h. Post-thaw assessments included routine sperm parameters, sperm DNA fragmentation index (DFI) evaluated by sperm chromatin dispersion assay, reactive oxygen species (ROS) levels measured via flow cytometry, RSV and nuclear factor erythroid 2-related factor 2 (NRF2) interactions examined by molecular docking and cellular thermal shift assay (CETSA), NRF2 protein contents analyzed by immunofluorescence and Western blotting, mRNA levels of NRF2 and downstream antioxidant proteins Heme Oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1) quantified by qRT-PCR and effects of NRF2 inhibitor ML385 on sperm parameters. Results:Compared with fresh samples, conventional cryopreservation significantly reduced sperm motility (all P<0.001). The addition of 10 -? mol/L RSV significantly improved the percentage of forward motile sperm after freezing (26.98%±8.98% vs. 19.61%±8.03%, P<0.001) while reducing DFI (9.84%±3.81% vs. 15.06%±4.22%, P<0.001) and ROS levels ( P<0.001) compared with the post-freezing group without the addition of RSV. Both molecular docking analysis and CETSA confirmed that RSV interacted with NRF2. Notably, sperm cryopreserved with 10 -? mol/L RSV exhibited significantly higher contents of NRF2 and its downstream effectors HO-1 and NQO1 compared with the post-freezing group without the addition of RSV (all P<0.001). This protective effect was markedly attenuated by co-treatment with the NRF2 inhibitor ML385, as evidenced by a significant decline in sperm motility ( P<0.001). Conclusion:RSV exerts cryoprotective effects likely through NRF2-mediated antioxidant pathways, reducing oxidative stress and enhancing post-thaw sperm quality.

Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T _NANC), time to symptom improvement (T _SI), and time of hospital stay (T _HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T _NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.

We analyzed the epidemiological features and spatial distribution characteristics of human brucellosis in Jining city from 2014 to 2023,to provide a reference for further development of targeted prevention and control strategies and measures.Descrip-tive epidemiological methods were used to analyze the epidemiological characteristics of brucellosis cases in Jining from 2014 to 2023.The spatial regional correlation of brucellosis incidence in Jining and the clustering patterns of local areas were studied through spatial autocorrelation analysis with townships as the basic unit.A total of 3 520 cases of brucellosis were reported in Jining from 2014 to 2023,and the average annual incidence rate was 4.23/100 000,thus indicating a fluctuating trend overall.Reported cases peaked from March to August,and a sex ratio of 2.71 males to 1 female was observed.The 40-59 year age group had the most reported cases(50.39％).The incidence of brucellosis in Jining showed an imbalanced spatial distribution.Brucellosis incidence showed a spatially clustered distribution(Moran's I>0,P<0.05).Hotspots were distributed primarily in Sishui,Qufu,and Zoucheng.A total of one class Ⅰ clustering area and one class Ⅱ clustering area were detected in the spatial and temporal scans,and were located in Sishui,Qufu,and Liangshan county.After pathogenic AMOS-PCR typing analysis,64 Brucella isolates collected from Jinan City from 2022 to 2024 were all of the sheep strain,and sheep biovar 3 was predominant(70.31％).In 2014-2023,although Jining City experienced a high incidence of brucellosis,a downward trend was observed.Brucellosis showed a spatial clustering pattern concentrated in the northeastern region.Therefore,awareness and education must be strengthened among brucellosis practitioners in cluster areas,to en-hance case surveillance,improve the level of protection,and achieve early detection and treatment.

Objective:To investigate the current status and related factors of antiviral treatment for chronic hepatitis B virus (HBV) infection in rural communities in China.Methods:In 2023, 866 chronic HBV-infected individuals from rural communities in Zhangqiu District, Jinan City, were included in the study. Basic information, disease status and antiviral treatment conditions of the infected individuals were collected through questionnaires, specimen collection and laboratory tests. Univariate and multivariate logistic regression were used to analyze the related factors of the antiviral treatment rate of those who met the indications for hepatitis B antiviral treatment.Results:The median age ( Q1, Q3) of subjects was 56 (48, 66) years old, among which 436 (50.4%) were males. There were 712 (82.2%) individuals who met the indications for hepatitis B antiviral treatment, and 110 individuals received antiviral treatment with a rate of 15.5%. Multivariate logistic regression analysis showed that compared with males, families with an average monthly income per capita of <1 000 yuan, no alcohol consumption, no smoking, and a family history of HBV infection, females ( OR=4.66, 95% CI: 2.88-7.53), families with an average monthly income per capita of 1 000-1 999 yuan ( OR=1.64, 95% CI: 1.00-2.68) and ≥2 000 yuan ( OR=2.78, 95% CI: 1.54-5.03), alcohol consumption ( OR=6.42, 95% CI: 2.80-14.7), smoking ( OR=1.98, 95% CI: 1.04-3.77), and no family history of HBV infection ( OR=1.90, 95% CI: 1.16-3.09) had a lower antiviral treatment rate for chronic HBV infection. Conclusion:The antiviral treatment rate of chronic HBV-infected individuals in rural communities of Zhangqiu District, Jinan City is low, and the related factors are female, high monthly income per capita, alcohol consumption, smoking, and no family history of HBV infection.

Objective:To evaluate the role of hippocampal activating transcription factor 5 (ATF5) in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR) in mice.Methods:This study was conducted in 2 parts. Experiment Ⅰ Twenty-four SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups ( n=12 each) using a random number table method: sham operation group (S1 group) and neuropathic pain group (NP group). Neuropathic pain was induced by chronic constriction injury to the sciatic nerve. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model and at 7, 14, 21 and 28 days after developing the model. Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model. After the end of the novel object recognition test, mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5 (by Western blot) and the expression of ATF5 in neurons, microglia and astrocytes (by immunofluorescence double staining). Experiment Ⅱ Thirty-six SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S2 group), neuropathic pain + ATF5 up-regulation group (NA group), and neuropathic pain + empty virus group (NE group). On day 14 after developing the model, a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region. The MWT and TWL were measured at days 28 and 35 after developing the model. The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function. After the end of the behavioral test, mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins (Lon protease [LONP1] and heat shock protein 60 [HSP60]) by Western blot. Results:Experiment Ⅰ Compared with S1 group, no statistically significant change was found in the MWT and TWL before developing the model ( P>0.05), the MWT and TWL were significantly decreased on days 7, 14, 21 and 28 after developing the model, the discrimination index (DI) was decreased at day 31 after developing the model, the expression of ATF5 was down-regulated, the expression of ATF5 in neurons was down-regulated ( P<0.05), and no statistically significant change was found in the expression of ATF5 in mircrolia and astrocytes in NP group ( P>0.05). Experiment Ⅱ Compared with S2 group, the MWT and TWL were significantly decreased on days 28 and 35 after developing the model in NE group and NA group, DI was decreased, and the expression of ATF5, LONP1 and HSP60 was down-regulated in NE group ( P<0.05), and no significant change was found in NA group ( P>0.05). Compared with NE group, no significant change was found in the MWT and TWL in NA group ( P>0.05), DI was significantly increased, and the expression of ATF5, LONP1 and HSP60 was up-regulated in NA group ( P<0.05). Conclusions:Down-regulated ATF5 in the hippocampus is involved in the process of cognitive impairment caused by neuropathic pain, and the mechanism may be related to the inhibition of mtUPR.

Objective To investigate the dynamic functional connectivity differences between insomnia patients and healthy controls in triple brain networks[the significant network(SN),the default mode network(DMN),and the executive control network(ECN)]using functional magnetic resonance imaging,and uncover their associations with cognitive ability.Methods Dynamic functional connectivity analysis was performed on functional magnetic resonance imaging data from 40 insomnia patients and 40 healthy controls.The changes in dynamic functional connectivity values were studied for SN,DMN,ECN[including the left executive control network(LECN)and the right executive control network(RECN)];the similarities and differences in time characteristic indicators such as time score,average dwell time,and conversion rate were explored;and their associations with clinical information were analyzed.Results The SN-LECN and DMN-RECN dynamic functional connectivity was significantly higher in insomnia patients than in healthy controls(P=0.013,0.047),while the RECN-LECN and RECN internal functional connectivity strength was lower in insomnia patients than in healthy controls(P<0.001).Additionally,the fractional time in state 2 in insomnia group was significantly higher than that in healthy controls(P<0.001),and it was positively correlated with the Pittsburgh sleep quality index(r=0.524,P=0.001).Conclusion Insomnia patients exhibit significant abnormalities in triple brain network dynamic functional connectivity,which may be related to abnormalities in cognitive control and sensory processing in insomnia patients.These findings provide a new perspective for further research on the neural mechanisms and potential intervention strategies for insomnia.

Objective To evaluate the predictive significance of contrast-enhanced ultrasound parameters in combination with serum levels of chemokine ligand 9(CXCL9)and insulin-like growth factor-binding protein 3(IGFBP-3)for the recurrence of hepatocellular carcinoma following interventional therapy.Methods A total of 212 patients with liver cancer who received interventional therapy at the hospital between June 2022 and Novem-ber 2023 were selected and categorized into Group A(61 cases)and Group B(151 cases)based on whether tumor recurrence occurred within one year following the intervention.Multivariate logistic regression analysis was con-ducted to identify potential influencing factors associated with post-treatment recurrence.The predictive perfor-mance of contrast-enhanced ultrasound parameters combined with serum biomarkers was evaluated using receiver operating characteristic(ROC)curve analysis.Results The levels of several factors including a tumor with a maximum diameter exceeding 3 cm,presence of microvascular invasion,high peak intensity(PI),short time to peak(TTP),short arrival time(AT),elevated serum CXCL9,and decreased serum IGFBP-3 were identified as indepen-dent risk factors for recurrence in patients with hepatocellular carcinoma following interventional therapy(P<0.05).The area under the curve(AUC)for the combination of PI,TTP,AT,along with serum levels of CXCL9 and IGFBP-3,in predicting recurrence after interventional therapy was significantly higher than that of individual fac-tors alone(P<0.05).Conclusions The risk of recurrence in patients undergoing interventional therapy for liver cancer is substantial,and it is influenced by a range of independent risk factors.Combining contrast-enhanced ul-trasound parameters with serum levels of CXCL9 and IGFBP-3 may enhance the predictive accuracy for tumor re-currence in these patients.

Objective To analyze the results of newborn screening for galactocerebrosidase(GALC)gene and enzyme activity and pro-vide a basis for the clinical diagnosis and genetic counseling of Krabbe disease.Methods The dried blood samples on filter paper from 12 744 newborns born at Nanjing Women and Children's Healthcare Hospital from March 18,2022 to December 31,2022 were collect-ed.The pathogenic variant sites of GALC gene were detected using the chip capture next-generation sequencing technology and Sanger sequencing was used for family validation.The tandem mass spectrometry was used to determine the enzyme activity of GALC in dried blood spots.The comparison of GALC enzyme activity between the carriers with GALC variants and negative group was conducted using the independent sample t-test.Results Among the 12 744 newborns,315 were identified as the carriers with GALC variants,with a carrier rate of 1/40.The most common variant was c.1901T＞C(269 cases,1/47),followed by c.1592G＞A(12 cases,1/1 062).Four newborns(P1-P4)were found to have two pathogenic variant sites and 2 cases were homozygous variants of c.1901T＞C.Family validation showed that the two variant sites in the four newborns were inherited from their parents,leading to a diagnosis of Krabbe dis-ease.Meanwhile,the family validation of the potential P2 patient's sister revealed that they had the same genotype,and the diagnosis of Krabbe disease was made.A preliminary threshold of 0.42 μmol/(L·h)for the enzyme activity of GALC detected by the tandem mass spectrometry was established based on 200 newborn samples.The GALC enzyme activities of 4 children with Krabbe disease were 0.74,0.21,0.17,and 0.29 μmol/(L·h),respectively.The GALC enzyme activity of the P2 patient's sister was 0.28 μmol/(L·h).Except for the P1 patient,the GALC enzyme activities of the P2,P3,and P4 patients and the P2 patient's sister were reduced,indica-ting a positive result.Conclusion The carrier rate of pathogenic variants in the GALC gene is relatively high,with the hotspot mutation being c.1901T＞C.The estimated prevalence of Krabbe disease is 1/3 186.The screening strategy using genetic testing as the primary screening and enzyme activity testing as the second screening can effectively improve the diagnostic efficiency of Krabbe disease,provi-ding a reference for the clinical diagnosis and genetic counseling of Krabbe disease.