ObjectiveTo investigate the effect and mechanism of transplantation of human umbilical cord mesenchymal stem cell （hUC-MSC） with overexpression of the Numb gene in the treatment of cholestatic liver fibrosis （CLF）. MethodsThe technique of lentiviral transfection was used to induce the overexpression of the Numb gene in hUC-MSC （hUC-MSC^Numb-OE）， and hUC-MSC transfected with empty vector （hUC-MSC^OE-EV） was used as negative control. Bile duct ligation （BDL） was performed to establish a rat model of CLF， and then the rats were randomly divided into BDL group， hUC-MSC group， hUC-MSC^OE-EV group， and hUC-MSC^Numb-OE group， while a sham-operation group was also established. The rats in the intervention groups were given a single splenic injection of the corresponding cells after BDL， and samples were collected at the end of week 4. Related indicators were measured， including serum biochemistry， liver histopathology， the content of hydroxyproline （Hyp） in the liver， hepatic stellate cell activation， ductular reaction， liver regeneration， and the expression levels of key molecules in the Numb-p53 signaling axis. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the BDL group， the hUC-MSC group and the hUC-MSC^OE-EV group had significant reductions in the levels of serum biochemical parameters （aspartate aminotransferase， gamma-glutamyl transpeptidase， total bile acid， total bilirubin， and direct bilirubin）， liver fibrosis markers （the content of Hyp and the expression levels of alpha-smooth muscle actin， tumor necrosis factor-α， and transforming growth factor-beta 1）， and ductular reaction markers （the expression levels of CK7 and CK19） （all P <0.05）， and compared with the hUC-MSC^OE-EV group， the hUC-MSC^Numb-OE group had significantly greater improvements in the above indicators （all P <0.05）. In addition， compared with the hUC-MSC^OE-EV group， the hUC-MSC^Numb-OE group had significant improvements in the expression levels of liver regeneration-related markers （albumin and hepatocyte nuclear factor 4α） and the molecules associated with the Numb-p53 signaling axis （Numb， pNumb， Mdm2， and p53） （all P <0.05）. ConclusionOverexpression of the Numb gene can enhance the therapeutic effect of hUC-MSC on CLF， possibly by activating the Numb-PTB^L-p53-HNF4α axis， promoting the hepatic differentiation of hUC-MSCs and subsequently enhancing liver regeneration.

Objective:To investigate the spatial and temporal distribution characteristics of brucellosis in Heilongjiang Province, Jilin Province and Inner Mongolia Autonomous Region (hereafter referred to as the Hei-Ji-Meng province and region), and study the influencing factors of brucellosis incidence.Methods:Using a retrospective analysis method, the data of brucellosis incidence in 22 prefecture-level cities in the Hei-Ji-Meng province and region from 2014 to 2018, as well as the data of natural environmental factors (precipitation, temperature, relative humidity, solar radiation, wind speed, vegetation index) and social environmental factors (livestock breeding, livestock and poultry meat production, cultivated land area) were collected and integrated. The spatial autocorrelation model and spatial Dubin model were used to analyze the spatial and temporal distribution rule of brucellosis and the impact of environmental factors on the incidence of brucellosis in prefecture-level cities.Results:From 2014 to 2018, the incidence of brucellosis in the Hei-Ji-Meng province and region was 18.77/100 000, among which the incidence in Inner Mongolia Autonomous Region was 33.45/100 000, followed by Heilongjiang Province (13.67/100 000) and Jilin Province (6.57/100 000). Among all prefecture-level cities, the brucellosis incidence was relatively higher in Hinggan League of Inner Mongolia Autonomous Region (69.10/100 000). Except 2016, the brucellosis incidence in other years had a positive spatial correlation (global Moran's I > 0, P < 0.05). The high-high concentration aggregation pattern was mainly concentrated in Ulanqab City, Xilingol League, Chifeng City, Tongliao City, Hinggan League, Hohhot City, and Hulunbuir City of Inner Mongolia Autonomous Region. Among the environmental factors related to brucellosis, cultivated land area and sheep breeding density had a positive effect on brucellosis incidence in the local area, with the effect intensity of 0.39 and 0.32, respectively ( P < 0.001). The average humidity from April to July had a negative effect on the brucellosis incidence in the local area, with the effect intensity of - 0.22 ( P = 0.023). The indirect effect of the number of pigs slaughtered on the brucellosis incidence in adjacent areas was the most significant, and the effect intensity was 2.15 ( P < 0.001). Conclusions:Among the Hei-Ji-Meng province and region, the annual average incidence rate of brucellosis in Inner Mongolia Autonomous Region is relatively higher, and the high-high concentration areas are mainly concentrated in this region. Among the environmental factors, the cultivated land area, sheep breeding density, the average humidity from April to July and the number of pigs slaughtered have an impact on brucellosis incidence.

Objective:To investigate the efficacy and safety of percutaneous trans-hepatic choledochoscopic lithotripsy (PTCSL) in the treatment of recurrent hepatobiliary calculi under enhanced recovery after surgery (ERAS) mode.Methods:Clinical data of 88 patients with recurrent hepatobiliary calculi, who were treated with PTCSL at Zhengzhou Central Hospital Affiliated to Zhengzhou University and the First Affiliated Hospital of Xi'an Jiaotong University between June 2022 and June 2024 were retrospectively analyzed, including 34 males and 54 females, aged (52.0±13.8) years. The scheme includes preoperative education, prophylactic antibiotic application, ensuring the quality of surgery, early postoperative feeding and activity, etc. The operation can be divided into two fashions: percutaneous transhepatic cholangial drainage and PTCSL, which can be completed in one stage (one-stage expansion method) or in two stages (staged expansion method). Clinical data such as gender, age, operative time, intraoperative blood loss, residual stone, and surgical complications were recorded.Results:All 88 patients underwent PTCSL under ERAS mode successfully, including 52 cases using one-stage expansion method and 36 cases using staged expansion method. The operative time was (53±20) min, the intraoperative blood loss was (9.7±3.8) ml, the postoperative hospital stay was (3.6±1.7) d, and the hospitalization cost was (17 500±4 700) yuan. Sixty-nine patients (78.4%, 69/88) had one-time stone removal in the first PTCSL. A total of 19 cases of residual stones were managed again by percutaneous sinus soft choledochoscopy, of which 12 cases were managed by one-time choledochoscopy, five cases by two-time choledochoscopy, and two cases by three-time choledochoscopy. The rate of residual stone was significantly higher in one-stage expansion method compared to staged expansion method [28.8% (15/52) vs. 11.1% (4/36), P=0.040]. No death, conversion to open surgery, or severe complications such as intra-abdominal hemorrhage or bile leakage occurred in the patients. No residual stones or recurrence were found during the follow-ups of (7.5±2.1) months after discharge. Conclusion:PTCSL under ERAS mode is safe and effective in the treatment of recurrent hepatobiliary calculi.

Objective To investigate the inhibitory effects of formononetin on sortase A(SrtA)of Staphylococcus epidermidis(S.epi-dermidis)and its biofilm formation.Methods Biofilm-producing S.epidermidis standard strain RP62A and biofilm-producing S.epider-midis clinical strains were collected.The minimal inhibitory concentration(MIC)of formononetin against S.epidermidis was investigated using the two-fold dilution method.The inhibitory effects of different concentrations of formononetin on the growth of S.epidermidis in vitro were studied by growth curve analysis using microplate reader.The cytotoxicity of formononetin was measured by cell culture.The biocompatibility of formononetin was studied by hemolysis assay.The inhibitory effect of formononetin on the activity of SrtA was deter-mined by fluorescence resonance energy transfer(FRET)assay.The effect of formononetin on the ability of S.epidermidis binding fi-bronectin was assessed by fibronectin binding assay.The effect of formononetin on the anchoring of serine-rich repeat proteins(SRRPs)in S.epidermidis was assessed by immunofluorescence staining.The effect of formononetin on the biofilm formation of S.epidermidis was measured by crystal violet staining.Results The MICs of formononetin against S.epidermidis were greater than 512 μg/mL.Compared with the control group,32 μg/mL of formononetin did not significantly affect the growth of S.epidermidis or reduce the viability of hu-man aortic endothelial cells.The hemolysis rate of human red blood cells was less than 5%.The inhibition rate of SrtA activity was ap-proximately 53.8%.The binding capacity of S.epidermidis to fibronectin was reduced by approximately 45.4%.Microscopic observation revealed a significant reduction in the anchoring of SRRP in S.epidermidis,and the formation of biofilm of S.epidermidis was inhibited by approximately 41.7%.Conclusion Formononetin inhibits the activity of SrtA in S.epidermidis,reduce its fibronectin-binding ca-pacity and biofilm formation,and thus represents a promising candidate for the prevention of S.epidermidis infections.

Objective:To explore the effects of peripheral nerve injury(PNI)on neuropathic pain(NP)and memo-ry function in mice,as well as the activation of neurons in the paraventricular nucleus(PVT)of the thalamus,so as to provide a basis for studying the relationship between NP and memory impairment.Methods:Twenty one 8-week-old male C57 BL/6J mice were randomly divided into sham group and experimental group,and the routine spared nerve inju-ry(SNI)was constructed in the mice of experimental group.The pain behavior and memory impairment of mice after SNI were evaluated with hot plate and eight-arm maze behavioral tests.Pearson correlation analysis was performed to an-alyze the correlation between pain behavior and memory impairment.The c-FOS expression in PVT was detected with immuno-staining.Results:Compared with the sham group,the heat pain threshold of mice in the experimental group was significantly reduced(P＜0.001).The results of the eight-arm maze test showed that the total rest time was signifi-cantly increased(P＜0.001),and the working memory error was increased from 1 to 4 days after SNI(P＜0.01).Correlation analysis indicated that early working memory errors were negatively correlated with heat pain threshold after SNI(P＜0.001).The immunofluorescence revealed that the number of c-FOS positive cells in PVT increased signifi-cantly(P＜0.001).Conclusion:SNI can cause abnormal pain behavior and memory impairment in mice,and cause neuronal activation in PVT.This study provides a basis for neurons in PVT to participate in the regulation of memory impairment in the context of NP.

Objective:To explore the effects of peripheral nerve injury(PNI)on neuropathic pain(NP)and memo-ry function in mice,as well as the activation of neurons in the paraventricular nucleus(PVT)of the thalamus,so as to provide a basis for studying the relationship between NP and memory impairment.Methods:Twenty one 8-week-old male C57 BL/6J mice were randomly divided into sham group and experimental group,and the routine spared nerve inju-ry(SNI)was constructed in the mice of experimental group.The pain behavior and memory impairment of mice after SNI were evaluated with hot plate and eight-arm maze behavioral tests.Pearson correlation analysis was performed to an-alyze the correlation between pain behavior and memory impairment.The c-FOS expression in PVT was detected with immuno-staining.Results:Compared with the sham group,the heat pain threshold of mice in the experimental group was significantly reduced(P＜0.001).The results of the eight-arm maze test showed that the total rest time was signifi-cantly increased(P＜0.001),and the working memory error was increased from 1 to 4 days after SNI(P＜0.01).Correlation analysis indicated that early working memory errors were negatively correlated with heat pain threshold after SNI(P＜0.001).The immunofluorescence revealed that the number of c-FOS positive cells in PVT increased signifi-cantly(P＜0.001).Conclusion:SNI can cause abnormal pain behavior and memory impairment in mice,and cause neuronal activation in PVT.This study provides a basis for neurons in PVT to participate in the regulation of memory impairment in the context of NP.

The Blow-Fill-Seal(BFS)technology is a highly automated aseptic manufacturing process that significantly reduces contamination risks by integrating container blowing,filling,and sealing operations.First,this article introduced the core technical features of BFS equipment and its advantages in aseptic production,followed by a comparative analysis of regulatory requirements for BFS technology across different jurisdictions.Finally,in light of pharmaceutical industry trends,it explored the future development prospects of BFS technology in intelligent manufacturing and high-barrier material applications,while examining its potential value in personalized medicine and specialized drug production.

Objective Biomarkers associated with liver cancer CD8+T cells identified and prognostic models established by single-cell RNA sequencing and Bulk RNA sequencing.Methods Single-cell datasets of liver cancer were downloaded from the GEO database and differentially expressed genes in CD8+T cells between patients and controls were extracted by single-cell RNA sequencing.Gene expression profiling data and clinical data of liver cancer were downloaded from the TCGA database,and modular genes with relevance to CD8+T cells were screened using the CIBERSORT algorithm and WGCNA technology.Differential genes and modular genes were taken as intersecting genes and subjected to GO and KEGG analyses,and a prognostic model was established by applying univariate COX regression analysis and LASSO algorithm.The predictive effectiveness of the model in both internal and external datasets is validated by K-M and ROC curves.The effectiveness of the model's prediction in both internal and external datasets was validated by K-M and ROC curves.High-and low-risk groups were divided according to the median value of the risk score,and the distribution of infiltrating immune cells and tumor mutations between high-and low-risk groups were analyzed.Results A prognostic model with nine genes was constructed,and the K-M and ROC curves showed that the model had good predictive ability in both the internal and external datasets,and there were significant differences in the distribution of infiltrating immune cells and gene mutations in the high-and low-risk groups.Conclusion In this study,a novel prognostic model based on CD8+T cells was developed using bioinformatic method in combination with single-cell RNA sequencing and Bulk RNA sequencing technologies,which provides a reliable theoretical basis for prognostic improvement and survival prediction of liver cancer patients.

The Blow-Fill-Seal(BFS)technology is a highly automated aseptic manufacturing process that significantly reduces contamination risks by integrating container blowing,filling,and sealing operations.First,this article introduced the core technical features of BFS equipment and its advantages in aseptic production,followed by a comparative analysis of regulatory requirements for BFS technology across different jurisdictions.Finally,in light of pharmaceutical industry trends,it explored the future development prospects of BFS technology in intelligent manufacturing and high-barrier material applications,while examining its potential value in personalized medicine and specialized drug production.

Purpose To investigate the feasibility of a deep learning framework to automatically analyze echocardiographic color Doppler videos in detecting valvular regurgitation.Materials and Methods This study retrospectively collected echocardiographic images of 1 109 patients with valvular regurgitation in the Fourth Medical Center of PLA General Hospital,from June 2015 to September 2019 as the training and validation sets.A prospective continuous collection of 1 562 echocardiography images was used as the test set in the Fourth Medical Center of PLA General Hospital from May 13 to June 13,2023,including 378 cases of mitral regurgitation and 223 cases of aortic regurgitation.This study developed deep learning networks to establish view classification model and valvular regurgitation recognition model,including the efficiency of section classification of deep learning models.Results The deep learning view classification model in this study could automatically identify two views for diagnosing mitral regurgitation and aortic regurgitation.The recognition accuracy for the parasternal long axis color Doppler view and the apical four chamber mitral color Doppler view was 1.00 and 0.93,respectively.The sensitivity,specificity,accuracy and area under the curve of the deep learning model for diagnosing mitral regurgitation were 0.847,0.852,0.849 and 0.930,respectively.The sensitivity,specificity,accuracy and area under the curve of the deep learning model in diagnosing aortic regurgitation were 0.857,0.861,0.859 and 0.940,respectively.Conclusion Deep learning algorithms can automatically identify valvular regurgitation and have the potential to become a screening tool for valvular heart disease.