ObjectiveTo investigate the potential mechanisms and pathways through which Gandouling （GDL） exerts its effects in the treatment of liver fibrosis in Wilson disease. MethodsSixty male SD rats were randomly divided into six groups： the normal group， the model group， the GDL low-， medium-， and high-dose groups （0.24， 0.48， 0.96 g·kg^-1）， and the penicillamine group （90 mg·kg^-1）， with 10 rats in each group. A copper-loaded Wilson disease rat model was established by gavage administration of 300 mg·kg^-1 copper sulfate pentahydrate to all groups except the normal group. Hematoxylin-eosin （HE） staining and Masson staining were used to observe the pathomorphological changes in the liver. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of hyaluronic acid （HA）， laminin （LN）， procollagen type-Ⅲ peptide （PC-Ⅲ）， and collagen type-Ⅳ （C-Ⅳ）. Transmission electron microscopy was used to examine the ultrastructure of liver tissues. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression levels of liver tissues and serum exosomal long noncoding RNA H19 （LncRNA H19）， phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， and mammalian target of rapamycin （mTOR）. Western blot analysis was performed to assess the expression levels of PI3K， Akt， mTOR， and their phosphorylated forms， as well as autophagy-related proteins Beclin1 and microtubule-associated protein 1 light chain 3B （LC3-Ⅱ/LC3-Ⅰ） in liver tissues. Beclin1 and LC3-Ⅱ fluorescence signal intensity was observed by immunofluorescence. ResultsCompared with the normal group， the model group exhibited inflammatory cell infiltration in hepatocytes， unclear nuclear boundaries with cell cleavage and necrosis， and collagen fiber deposition around confluent areas. The levels of HA， LN， PC-Ⅲ， and C-Ⅳ were significantly elevated （P<0.01）. Transmission electron microscopy revealed an increased number of autophagic vesicles， with autophagic lysosomes exhibiting a single-layer membrane structure following degradation of most envelopes. Expression levels of Beclin1 and LC3-Ⅱ/LC3-Ⅰ were significantly increased （P<0.01）， and fluorescence signals of Beclin1 and LC3-Ⅱ were markedly enhanced. The protein expression levels of PI3K， Akt， mTOR， p-PI3K， p-Akt， and p-mTOR were reduced （P<0.01）， while LncRNA H19 expression was increased （P<0.01）， and mRNA expression levels of PI3K， Akt， and mTOR were decreased （P<0.01）. After treatment with GDL， the degree of liver fibrosis was significantly improved， with decreased levels of HA， LN， PC-Ⅲ， and C-Ⅳ. The number of autophagic vesicles was significantly reduced， and expression levels of Beclin1 and LC3-Ⅱ/LC3-Ⅰ proteins were lower （P<0.01）. The fluorescence signals of Beclin1 and LC3-Ⅱ weakened dose-dependently. The protein levels of PI3K， Akt， mTOR， p-PI3K， p-Akt， and p-mTOR were elevated （P<0.01）， while the expression level of LncRNA H19 was reduced （P<0.01）. Furthermore， the mRNA expression levels of PI3K， Akt， and mTOR increased （P<0.05， P<0.01）. ConclusionGDL may alleviate liver fibrosis and reduce liver injury by regulating the PI3K/Akt/mTOR autophagy signaling pathway via LncRNA H19.

Objective To investigate the value of serum thymidine kinase 1(TK1),hypoxia-inducible fac-tor-1α(HIF-1α)and squamous cell carcinoma antigen(SCC)in the diagnosis and prognosis of esophageal cancer.Methods A total of 105 patients with esophageal cancer treated in the Qinhuai Medical Zone of East-ern Theater General Hospital from February 2019 to October 2021 were selected as the study group,and 80 healthy subjects were selected as the control group during the same period.The serum levels of TK1,HIF-1αand SCC were compared between study group,control group and patients with different pathological charac-teristics.Patients with esophageal cancer were followed up for 3 years,and the overall survival(OS)and pro-gression-free survival(PFS)were recorded.Receiver operating characteristic curve was used to analyze the di-agnostic efficiency of serum TK1,HIF-1α and SCC combined detection for esophageal cancer,Pearson correla-tion analysis was used to analyze the correlation between the serum indicators,and Kaplan-Meier survival a-nalysis was used to analyze the OS and PFS of patients with different serum levels of TK1,HIF-1α and SCC.Multivariate COX regression was performed to analyze prognostic factors.Results Compared with the control group,the serum TK1,HIF-1α and SCC levels in the study group increased(P＜0.05).The area under the curve(AUC)of serum TK1,HIF-1α,SCC alone and combined for diagnosis of esophageal cancer were 0.893,0.744,0.841,0.922,respectively,and their combined diagnoses of esophageal cancer had the largest AUC.Se-rum TK1,HIF-1α and SCC in patients with different TNM stages and differentiation stages were significantly different(P＜0.05).Serum TK1,HIF-1α and SCC in patients with lymph node metastasis were higher than those without lymph node metastasis(P＜0.05).Serum TK1 was positively correlated with HIF-1α and SCC levels in patients with esophageal cancer(P＜0.05).The survival functions of OS and PFS in TK1,HIF-1αand SCC low expression group were better than those in high expression group(P＜0.05).Multivariate COX regression analysis showed that low differentiation and lymph node metastasis were independent risk factors for poor prognosis in esophageal cancer patients(P＜0.05).Conclusion Serum levels of TK1,HIF-1α and SCC are increased in patients with esophageal cancer,the combined diagnosis of the three is effective.The high expression of TK1 and HIF-1αand SCC will shorten OS and PFS.

Objective To investigate the relationship between serum microRNA(miRNA,miR)-9-5p,miR-21-5p and miR-206 and hemorrhagic transformation(HT)occurs and prognosis in patients with large ar-tery atherosclerotic cerebral infarction(ACI-LAA)after thrombolysis.Methods A total of 265 patients with ACI-LAA admitted to the hospital from April 2021 to November 2023 were selected as the research objects.According to whether HT occurred after intravenous thrombolysis,they were divided into non-HT group and HT group,and the expression levels of serum miR-9-5p,miR-21-5p and miR-206 were compared between the two groups.The general data of patients with ACI-LAA were collected.Univariate and multivariate Logistic regression analysis were used to analyze the influencing factors of HT occurs in patients with ACI-LAA after thrombolysis.The receiver operating characteristic(ROC)curve was used to analyze the efficacy of serum miR-9-5p,miR-21-5p and miR-206 expression levels in predicting HT occurs after thrombolysis in patients with ACI-LAA.The patients with ACI-LAA were followed up for 90 d.According to the modified Rankin scale(mRS)score,the patients were divided into a good prognosis group and a poor prognosis group,and the expression levels of serum miR-9-5p,miR-21-5p and miR-206 were compared between the two groups.Univa-riate and multivariate Logistic regression analysis were used to analyze the influencing factors of poor progno-sis in patients with ACI-LAA.ROC curve was used to analyze the efficacy of serum miR-9-5p,miR-21-5p and miR-206 expression levels in predicting poor prognosis of patients with ACI-LAA.Results HT occurred in 74(27.92％)of 265 patients with ACI-LAA after thrombolysis.Compared with the non-HT group,the National Institutes of Health Stroke Scale(NIHSS)score atadmission,the proportion of hypertension,the proportion of infarct size ≥10 cm2,and the expression levels of serum miR-21-5p and miR-206 were significantly in-creased in the HT group(P＜0.05).However,the expression level of serum miR-9-5p was decreased(P＜0.05).Hypertension,high NIHSS score at admission,the proportion of infarct size ≥10 cm2,high expression of miR-21-5p and miR-206 were risk factors for HT occurs in patients with ACI-LAA after intravenous thrombolysis,and high expression of miR-9-5p was a protective factor(P＜0.05).The ROC curve showed that the area under the curve(AUC)and 95％CI of single and combined detection of serum miR-9-5p,miR-21-5p and miR-206 for predicting HT occurs were 0.796(0.726-0.866),0.779(0.711-0.846),0.784(0.714-0.854),0.891(0.839-0.943),respectively.Among 265 patients with ACI-LAA,83 patients(31.32％)had poor prognosis at 90 d of follow-up.Compared with the good prognosis group,the age,NIHSS score at admis-sion,the proportion of HT,the proportion of infarct size ≥10 cm2,and the expression levels of serum miR-21-5p and miR-206 were increased(P＜0.05),and the expression level of serum miR-9-5p was decreased(P＜0.05)in the poor prognosis group.Advanced age,high NIHSS score at admission,HT,infarct size ≥10 cm2,high expression of miR-21-5p,and high expression of miR-206 were risk factors for poor prognosis of ACI-LAA patients,and high expression of miR-9-5p was a protective factor(P＜0.05).The ROC curve showed that The AUC(95％CI)of single and combined detection of serum miR-9-5p,miR-21-5p and miR-206 for predicting poor prognosis were 0.754(0.691-0.818),0.779(0.719-0.838),0.792(0.815-0.919),0.931(0.902-0.960),respectively.Conclusion Low expression of serum miR-9-5p,high expression of miR-21-5p and high expression of miR-206 are associated with the HT occurs and poor prognosis in patients with ACI-LAA after thrombolysis,and the combined detection of the three indicators has a high predictive value for the HT occurs and poor prognosis.

Objective:To analyze the characteristics of inhibitory control and cognitive flexibility in hearing-impaired children.Methods:From March to April 2023, a convenience sampling method was used to select 33 hearing-impaired children from a special education school in Meizhou City, Guangdong Province, and 35 normal-hearing children from two ordinary schools as participants. Inhibitory control and cognitive flexibility of the participants were assessed by the Flanker task and the dimensional change card sorting (DCCS) task. Statistical analysis was conducted using SPSS 26.0 software, and independent sample t-test was used to compare the differences in reaction time and accuracy rate between two groups of participants. Results:There were no significant differences in the Flanker task reaction time ((558.39±123.65) ms vs (566.11±118.20) ms) and accuracy rate((0.93±0.10) vs (0.96±0.04))between hearing-impaired children and normal-hearing children ( t=-0.295, -1.645, both P>0.05). The hearing-impaired children had significantly longer reaction time ((1 019.60±131.08) ms)than the normal-hearing children ((857.85±129.19) ms) ( t=4.046, P=0.001) in the DCCS task, while there was no statistically significant difference in the accuracy rate between hearing-impaired children (0.62±0.16) and normal-hearing children (0.57±0.15) ( t=-1.602, P>0.05). Conclusion:There is no difference in inhibitory control ability between hearing-impaired children and normal-hearing children, but the hearing-impaired children have a lag in cognitive flexibility.

Objective To compare image quality of 5.0T and 3.0T non-contrast MRI for displaying insulinoma.Methods Twelve patients with insulinoma were prospectively enrolled,and non-contrast abdominal T1WI,T2WI as well as diffusion-weighted imaging(DWI)were acquired using 5.0T and 3.0T MR scanners,respectively.The subjective scores of image quality of each sequence of 5.0T and 3.0T MRI,also of tumor-pancreas parenchyma contrast scores were compared.The signal-to-noise ratio(SNR)and contrast-to-noise ratio(CNR)of insulinomas were observed,and the displayed rate of insulinoma by each sequence and overall MRI were compared.Results The subjective scores of 5.0T T1WI and DWI were higher than those of 3.0T T1WI and DWI(both P＜0.05),but not significantly different between 5.0T and 3.0T T2WI(P=0.166).Furthermore,the tumor-pancreas parenchyma contrast score of 5.0T T1WI was higher than that of 3.0T T1WI(P=0.023),but not significantly different between 5.0T and 3.0T T2WI,nor between 5.0T and 3.0T DWI(both P＞0.05).SNR of insulinomas on 5.0T T2WI were higher than on 3.0T T2WI(P=0.015),however,no significant difference of SNR was found between 5.0T and 3.0T T1WI,nor between 5.0T and 3.0T DWI(both P＞0.05).CNR of insulinomas on all 5.0T MRI were not significantly different with those on 3.0T MRI(all P＞0.05).The displayed rate of insulinoma on 5.0T T1WI,T2WI and DWI was 100％(12/12),66.67％(8/12)and 83.33％(10/12),respectively,on 3.0TT1WI,T2WI and DWI was 75.00％(9/12),58.33％(7/12),66.67％(8/12),respectively.The overall displayed rate of insulinoma on 5.0T and 3.0T MRI was 100％(12/12)and 83.33％(10/12),respectively.Conclusion Compared with 3.0T MRI,5.0T MRI was superior for displaying insulinoma,hence being helpful for diagnosis.

Objective:To investigate the necessity of internal fixation of the fibular fracture in the treatment of degree Ⅱ ankle supination-external rotation injury by the Dias-Tachdjian classification in children.Methods:A retrospective study was conducted to analyze the data of 69 children with ankle fracture (degree Ⅱ ankle supination-external rotation injury by the Dias-Tachdjian classification) who had been treated at Department of Orthopedic Trauma, The People's Hospital of Weifang and Department of Pediatric Orthopedics, Tianjin Hospital from January 2015 to October 2021. There were 41 males and 28 females with an age of (10.1±1.4) years, and 37 left and 32 right sides affected. The patients were divided into 2 groups according to whether internal fixation of the fibular fracture was performed. Group A consisted of 21 cases with fibular internal fixation and group B of 48 cases without fibular internal fixation. The preoperative data, operation time and operation expenses were recorded and compared between the 2 groups. At the last follow-up, the anteroposterior and lateral radiographs of bilateral full length lower limbs and ankle joints were taken; the lateral distal tibial angle (LDTA) and the anterior distal tibial angle (ADTA) on the affected side, and the disparity between bilateral ankle tibiotalar angles were measured; ankle function was assessed according to the ankle-hindfoot score of American Association of Foot and Ankle Surgery (AOFAS); the occurrence of premature physeal closure (PPC) was recorded.Results:There was no significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). All patients were followed up for (19.5±4.1) months. At the last follow-up, the LDTA on the affected side was 89.6° (87.9°, 90.5°) in group A and 88.6°±1.9° in group B; the ADTA on the affected side was 80.9° (79.0°, 81.4°) in group A and 80.0° (78.6°, 81.2°) in group B; the disparity between bilateral ankle tibiotalar angles was 1.1°±0.5° in group A and 1.2°±0.5° in group B; the AOFAS ankle-hindfoot score was (89.5±5.2) points in group A and 89.0 (87.0, 92.0) points in group B. There was no statistically significant difference between the 2 groups in the above items ( P>0.05). The incidence of PPC was, respectively, 14.3% (3/21) and 14.6% (7/48) in groups A and B, showing no statistically significant difference ( P>0.05). Conclusions:In the treatment of degree Ⅱ ankle supination-external rotation injury in children, internal fixation of the fibular fracture has no significant effect on the imaging angles or the function of the ankle joint. After anatomic reduction of the distal tibia, the fibular fracture can be treated without internal fixation to reduce operational trauma, shorten operation time and reduce operation expenses.

Objective:To develop a wearable gait analysis system based on "Lab-in-Shoe" intelligent footwear for quantitative assessment of gait dysfunction in patients with ankle injuries.Methods:In this study, integration of inertial sensors and insole-type plantar pressure distribution sensors into footwear formed the hardware core of the "Lab-in-Shoe" intelligent footwear system. In terms of algorithms, acceleration data from the inertial sensors were integrated to obtain spatial parameters of gait. The insole-type plantar pressure sensors were employed to acquire the data concerning foot pressure distribution, as well as temporal parameters and mechanical parameters of gait, including support phase, swing phase, and zero velocity moments. To validate the accuracy of this system, 8 young and healthy participants [age: (25.6±1.3) years; height: (175.4±2.2) cm] were recruited for gait data collection in an optical motion capture laboratory. By comparing the gait data between the "Lab-in-Shoe" intelligent footwear system and the gold standard Vicon optical motion capture system, the effectiveness and reliability of the intelligent footwear system were respectively tested. Additionally, a calibration experiment was conducted for the sensing units of the plantar pressure sensors to examinate the accuracy of the pressure data.Results:The tested system accurately captured the following gait parameters of the participants: step length, step width, step frequency, walking speed, gait phase division, foot pressure distribution, and center of pressure curve, among other core spatiotemporal gait parameters. Moreover, the system demonstrated its ability to replicate the dual-foot posture during gait. Compared with the gold standard Vicon optical motion capture system through Bland-Altman, the Lab-in-Shoe smart shoe system showed stride length mean error within 3.12% (range: 2.76% to 4.24%) across 3 different walking speeds [slow speed (0.68±0.05) m/s, preferred speed (1.10±0.07) m/s, and fast speed (1.40±0.13) m/s]. 90% of the results fell within the 95% limits of agreement, indicating good consistency. The intraclass correlation coefficients (ICC) for stride parameters within the slow, preferred, and fast walking speed groups were 0.93, 0.917, and 0.893, respectively, indicating good reliability. The calibration data of multiple sensor units from the plantar pressure sensors all fell within the 95% linear regression range, with a correlation coefficient of r=0.949 ( P<0.05). The plantar pressure data collected by the intelligent footwear system presented a distinct bimodal characteristic. Conclusions:The "Lab-in-Shoe" smart shoe system developed by our institute is capable of collecting and calculating gait parameters conveniently and quickly, and demondtrates good reliability and validity across different walking speeds. Therefore, it is valueable for large-scale gait data collection in a clinical setting.

Objective:To identify early potential risk factors for sepsis-associated liver injury and to provide a reference for early clinical identification and intervention.Methods:The clinical data of septic patients admitted to the intensive care unit (ICU) in the Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University from March 2020 to April 2023 were retrospectively analyzed. Patients with sepsis were categorized into the liver injury group and the non-liver injury group according to whether liver injury occurred or not, univariate and multivariate logistic regression analyses were used to explore the risk factors for SALI, receiver operating characteristic (ROC) curve analysis was used to assess its predictive effect for SALI, and performed subgroup analyses basing on the cut-off point.Results:Among 530 eligible patients, 403 patients were included. The incidence of liver injury was 39.45% in 159 cases with liver injury and 244 cases without liver injury. Multivariate logistic regression analysis showed that serum prealbumin, lactate and lactate dehydrogenase were independent risk factors for SALI. ROC curve analysis showed that all single indicators had some predictive value for SALI, the area under the curve was prealbumin (AUC: 0.752, 95% CI: 0.703-0.801), lactate (AUC: 0.679, 95% CI: 0.627-0.732), lactate dehydrogenase (AUC: 0.664, 95% CI: 0.611-0.718), respectively, The AUC for predicting SALI by lactate/prealbumin ratio (L/P) and lactate dehydrogenase/prealbumin ratio were 0.808 (95% CI: 0.766-0.850) and 0.795 (95% CI: 0.750-0.840), respectively, with the best efficacy of L/P in predicting SALI. Subgroup analyses showed that the incidence of liver injury was significantly higher in septic patients with L/P ≥0.23 than that in septic patients with L/P <0.23, at the same time, the acute physiology and chronic health evaluation II score, shock probability, and hospital mortality rate also increased accordingly, the differences were all statistically significant (all P < 0.001). Conclusions:L/P is early independent risk factor of SALI, for sepsis patients with L/P≥0.23 should be alerted to the development of liver injuryis.

Infertility affects an estimated 10 to 15 percent of couples worldwide,with approximately half of the cases attributed to male-related issues.Most men diagnosed with infertility exhibit symptoms such as oligospermia,asthenospermia,azoospermia,and compromised sperm quality.Spermatogenesis is a complex and tightly coordinated process of germ cell differentiation,precisely regulated at transcriptional,posttranscriptional,and translational levels to ensure stage-specific gene expression during the development of spermatogenic cells and normal spermiogenesis.N6-methyladenosine(m6A)stands out as the most prevalent modification on eukaryotic mRNA,playing pivotal roles in various biological processes,including mRNA splicing,transportation,and translation.RNA methylation modification is a dynamic and reversible process primarily mediated by"writers",removed by"erasers",and recognized by"readers".In mammals,the aberrant methylation modification of m6A on mRNA is associated with a variety of diseases,including male infertility.However,the precise involvement of disrupted m6A modification in the pathogenesis of human male infertility remains unresolved.Intriguingly,a significant correlation has been found between the expression levels of m6A regulators in the testis and the severity of sperm concentration,motility,and morphology.Aberrant expression patterns of m6A regulatory proteins have been detected in anomalous human semen samples,including those of oligospermia,asthenozoospermia,and azoospermia.Furthermore,the examination of both sperm samples and testicular tissues revealed abnormal mRNA m6A modification,leading to reduced sperm motility and concentration in infertile men.Consequently,it is hypothesized that dysregulation of m6A modification might serve as an integral link in the mechanism of male infertility.This paper presents a comprehensive review of the recent discoveries regarding the spatial and temporal expression dynamics of m6A regulators in testicular tissues and the correlation between deregulated m6A regulators and human male infertility.Previous studies predominantly utilized constitutive or conditional knockout animal models for testicular phenotypic investigations.However,gene suppression in additional tissues could potentially influence the testis in constitutive knockout models.Furthermore,considering the compromised spermatogenesis observed in constitutive animals,distinguishing between the indirect effects of gene depletion on testicular development and its direct impact on the spermatogenic process is challenging,due to their intricate relationship.Such confounding factors might compromise the validity of the findings.To address this challenge,an inducible and conditional gene knockout model may serve as a superior approach.To date,nearly all reported studies have concentrated solely on the level changes of m6A and its regulators in germs cells,while the understanding of the function of m6A modification in testicular somatic cells remains limited.Testicular somatic cells,including peritubular myoid cells,Sertoli cells,and Leydig cells,play indispensable roles during spermatogenesis.Hence,comprehensive exploration of m6A modification within these cells as an additional crucial regulatory mechanism is warranted.In addition,exploration into the presence of unique methylation mechanisms or m6A regulatory factors within the testes is warranted.To elucidate the role of m6A modification in germ cells and testicular somatic cells,detailed experimental strategies need to be implemented.Among them,manipulation of the levels of key enzymes involved in m6A methylation and demethylation might be the most effective approach.Moreover,comprehensive analysis of the gene expression profiles involved in various signaling pathways,such as Wnt/β-catenin,Ras/MAPK,and Hippo,in m6A-modified germ cells and testicular somatic cells can provide more insight into its regulatory role in the spermatogenesis process.Further research in this area could provide valuable insights for developing innovative strategies to treat male infertility.Finally,considering the mitigation impact of m6A imbalance regulation on disease,investigation concerning whether restoring the equilibrium of m6A modification regulation can restore normal spermatogenesis function is essential,potentially elucidating the pivotal clinical significance of m6A modulation in male infertility.

Infertility is a global health issue,and more and more people are hoping to have babies by means of assisted reproductive technology.However,there are still many challenges in fertilization and pregnancy outcomes.Sperm quality is a key factor affecting the success rate of assisted reproduction.Therefore,sperm quality screening is crucial for achieving breakthroughs in assisted reproduction technology.At present,with its capabilities in the field of image recognition,artificial intelligence(AI)is providing new ideas and methods for sperm screening.Various attempts have been made with AI-based models to evaluate indicators such as sperm morphology,DNA quality,and motility level,and some results have been achieved.Herein,we reviewed the application of AI in sperm quality analysis and selection,providing support for the future development of AI and the improvement in the fertilization rate and outcomes of assisted reproductive technology.