ObjectiveThis paper aims to investigate the traditional Chinese medicine syndrome types in patients with chronic hepatitis B （CHB） complicated by metabolic dysfunction-associated fatty liver disease （MAFLD） and explore the correlations between these syndrome types and clinical indicators， as well as ocular manifestation characteristics， thereby providing a reference for syndrome differentiation and treatment strategies in traditional Chinese medicine. MethodsGeneral data， information from the four diagnostic methods of traditional Chinese medicine， clinical indicators， and ocular manifestation data were collected from 506 patients with CHB complicated by MAFLD enrolled at the Public Health Clinical Center of Chengdu between June 2024 and December 2024. Cluster analysis， principal component analysis， and complex network models were employed to identify the distribution patterns of traditional Chinese medicine syndromes. Correlations between different syndrome types and clinical indicators， as well as ocular manifestation characteristics， were further analyzed. ResultsThe predominant syndromes identified in patients with CHB complicated by MAFLD were dampness and heat accumulation （51.58%）， liver depression with spleen deficiency （31.62%）， blood stasis obstructing collaterals （8.89%）， and Qi-Yin deficiency （7.91%）. No statistically significant differences were found among the four syndrome types in routine blood tests and liver function indicators. However， patients with the dampness and heat accumulation type exhibited significantly higher levels of total cholesterol （TC）， triglycerides （TG）， low-density lipoprotein cholesterol （LDL-C）， liver stiffness measurement （LSM）， controlled attenuation parameter （CAP）， and alpha-fetoprotein （AFP）， along with lower levels of high-density lipoprotein cholesterol （HDL-C）， compared with those with other syndrome types. Regarding ocular manifestations， the incidence of moon halo signs was significantly higher in patients with the blood stasis obstructing collaterals type than in those with other syndrome types. Additionally， the incidence in scleral zone 3 （corresponding to the large intestine） was higher in patients with the damp and heat accumulation type. ConclusionDampness and heat accumulation is the core syndrome type in patients with CHB complicated by MAFLD， commonly accompanied by spleen deficiency， liver depression， blood stasis， and Yin deficiency. A complex syndrome pattern characterized by a predominance of dampness and heat， along with a mixture of deficiency and excess， is formed. Different traditional Chinese medicine syndrome types are associated with distinct clinical indicators and ocular manifestation characteristics. Among them， patients with the dampness and heat accumulation type exhibit more pronounced metabolic disturbances and liver injury， whereas those with the blood stasis type show a higher incidence of moon halo signs. Abnormalities in scleral zone 3 are also more prevalent in patients with dampness and heat type.

ObjectiveThis paper aims to investigate the distribution patterns of traditional Chinese medicine syndromes in patients with chronic hepatitis B （CHB） comorbid with metabolically associated fatty liver disease （MAFLD） and analyze their correlation with clinical characteristics and the progression of liver fibrosis. MethodsA cross-sectional study method was employed， and 506 patients with CHB comorbid with MAFLD who attended the Hepatology Outpatient Department of Public Health Clinical Center of Chengdu from June 2024 to December 2024 were enrolled. General information， traditional Chinese medicine syndromes information， laboratory indicators， and imaging examination results were collected using case report forms （CRF）. Tongue images of patients were acquired using a tongue diagnosis instrument， and tongue feature parameters were extracted using computer image processing technology. Frequency analysis， factor analysis， and cluster analysis， and other methods were used to explore syndrome categories and distribution patterns. Non-parametric tests were used to compare the differences in clinical characteristics among different syndromes. Univariate and multivariate logistic regression analyses were performed to investigate the correlation between traditional Chinese medicine syndromes and the progression of liver fibrosis. ResultsThe main traditional Chinese medicine syndromes in patients with CHB comorbid with MAFLD were mainly dominated by damp-heat accumulation syndrome， liver stagnation and spleen deficiency syndrome， and phlegm-blood stasis syndrome， with damp-heat accumulation syndrome accounting for the highest proportion （41.89%）. Compared with those without damp-heat accumulation syndrome， patients with damp-heat accumulation syndrome had significantly lower tongue proper H value， tongue coating H value， and tongue coating a^* value （P<0.05）， significantly higher tongue coating b^* value （P<0.05）， significantly increased levels of white blood cell （WBC）， red blood cell （RBC）， hemoglobin （HGB）， and glucose （GLU）， increased CAP values （P<0.05）， a higher proportion of males （P<0.05）， and a younger age （P<0.05）. Univariate and multivariate logistic regression analyses show that age， hepatitis B surface antigen （HBsAg）， diabetes， and damp-heat accumulation syndrome are independent risk factors for liver fibrosis （P<0.05）， and that damp-heat accumulation syndrome is predominantly distributed in liver fibrosis stage F0-F1. ConclusionDamp-heat accumulation syndrome is a typical syndrome in patients with CHB comorbid with MAFLD， which is significantly associated with enhanced inflammatory response， metabolic disorders， and early liver fibrosis， and is a key link in disease progression. Clinical attention and early intervention are needed.

OBJECTIVES: To analyze the distribution of Y chromosome azoospermia factor (AZF) microdeletions and their association with testicular development in male pediatric patients with congenital reproductive system disorders, including hypospadias, cryptorchidism, and disorders of sex development (DSD). METHODS: A prospective cohort study was conducted on pediatric patients admitted to the Department of Urology of Shanghai Children's Hospital from November 2021 to December 2023. The observation group included boys with hypospadias, cryptorchidism, or DSD, while the control group comprised boys with phimosis, indirect inguinal hernia, or hydrocele. Blood samples were collected for AZF microdeletion analysis using multiplex PCR to detect 15 sequence-tagged sites. Testicular ultrasound was performed to record testicular position and volume. Propensity score matching (PSM) was used to balance the groups. After matching, testicular volume differences were assessed. Stratified analyses compared testicular volume among children with AZF microdeletions, the control group, and children without micro-deletions in observation group. RESULTS: A total of 493 children were enrolled (observation group: 463; control group: 30). No Y chromosome microdeletions were detected in the control group. Four boys in the observation group had AZF microdeletions: one with cryptorchidism (AZFc+AZFd), one with isolated hypospadias (AZFc), and two with DSD (one with AZFb+AZFc+AZFd and one with AZFa). Ultrasonography measured 888 testicles. After PSM, testicular volume was significantly smaller in the observation group than in the control group (P<0.01). Stratified analysis revealed that among children under 9 years, those with AZF microdeletions tended to be older but had smaller testicular volumes compared to the control group and those without microdeletions in the observation group, although differences were not statistically significant (all P>0.05). Among children over 9 years, ages were comparable, but children with AZF microdeletions had smaller testicular volumes than the other two groups (statistical analysis was not performed due to small sample size). CONCLUSIONS The prevalence of Y chromosome microdeletions is higher in male children with congenital reproductive system disorders compared to the general population, particularly in those with DSD. Hypospadias, cryptorchidism, DSD, and AZF microdeletions may be associated with delayed testicular development in these children.

Objective:To explore the possible mechanism of cognitive impairment in aged mice induced by sevoflurane and the role of the AMPK/GSK-3β/Nrf2 signaling pathway.Methods:Eighteen 16-month-old SPF male C57BL/6J mice were randomly assigned to either a control group or a sevoflurane group according to the random number table method( n=9 per group). Mice in the sevoflurane group were exposed to 3% sevoflurane for 2 h every day for three consecutive days.Cognitive function of mice was assessed by novel object recognition test and Morris water maze test.Golgi staining was used to analyze dendritic spine morphology and density in the hippocampus. Mitochondrial ultrastructure was examined by transmission electron microscopy. Oxidative stress in hippocampal tissue was evaluated by reactive oxygen species(ROS) and glutathione(GSH) assay kits. Western blot was performed to measure synaptic plasticity-related proteins and proteins involved in the AMPK/GSK-3β/Nrf2 signaling pathway, while Nrf2 expression was assessed by immunofluorescence. Statistical analysis was performed using GraphPad Prism 7 software with independent-samples t-test or Mann-Whitney U test for between-group comparisons. Results:(1) The novel object recognition test showed that the recognition index in the sevoflurane group was significantly lower than that in the control group ( Z=-3.256, P=0.001).The escape latency in the Morris water maze test was longer ((49.50±10.14) s, (18.62±6.59) s; t=-7.221, P<0.001) and the number of platform crossings was lower ( Z=-2.673, P=0.008) in the sevoflurane group compared with those of the control group. (2) Results from Western blot and Golgi staining showed that the expression levels of hippocampal synapse-related proteins in the sevoflurane group, including PSD95 ((0.38±0.07), (1.00±0.21); t=4.885) and SYN1 ((0.30±0.10), (1.00±0.10); t=8.575), were lower than those in the control group, as well as the number of dendritic spines ((10.3±2.5), (20.0±1.0); t=6.183)(all P<0.05). (3) Transmission electron microscopy showed mitochondrial damage in the hippocampus of the sevoflurane group, and the expression level of ROS ((3.05±0.90), (0.97±0.16); t=-4.555, P=0.004) was significantly higher than that of the control group, while the expression level of GSH ((0.71±0.07), (1.00±0.09); t=5.396, P=0.002) was significantly lower than that of the control group.(4) Western blot and immunofluorescence demonstrated that hippocampal p-AMPK, p-GSK-3β (Ser9), and HO-1 expression levels in the sevoflurane group were significantly lower than those in the control group ( t=2.845, 7.087, 4.551, all P<0.05), and Nrf2 fluorescence intensity was also markedly reduced ( P<0.05). Conclusion:The cognitive impairment induced by sevoflurane in aged mice may be caused by mitochondrial damage, oxidative stress response and synaptic damage, which maybe associated with the inhibition of the AMPK/GSK-3β/Nrf2 signaling pathway.

Objective:To explore the possible mechanism of cognitive impairment in aged mice induced by sevoflurane and the role of the AMPK/GSK-3β/Nrf2 signaling pathway.Methods:Eighteen 16-month-old SPF male C57BL/6J mice were randomly assigned to either a control group or a sevoflurane group according to the random number table method( n=9 per group). Mice in the sevoflurane group were exposed to 3% sevoflurane for 2 h every day for three consecutive days.Cognitive function of mice was assessed by novel object recognition test and Morris water maze test.Golgi staining was used to analyze dendritic spine morphology and density in the hippocampus. Mitochondrial ultrastructure was examined by transmission electron microscopy. Oxidative stress in hippocampal tissue was evaluated by reactive oxygen species(ROS) and glutathione(GSH) assay kits. Western blot was performed to measure synaptic plasticity-related proteins and proteins involved in the AMPK/GSK-3β/Nrf2 signaling pathway, while Nrf2 expression was assessed by immunofluorescence. Statistical analysis was performed using GraphPad Prism 7 software with independent-samples t-test or Mann-Whitney U test for between-group comparisons. Results:(1) The novel object recognition test showed that the recognition index in the sevoflurane group was significantly lower than that in the control group ( Z=-3.256, P=0.001).The escape latency in the Morris water maze test was longer ((49.50±10.14) s, (18.62±6.59) s; t=-7.221, P<0.001) and the number of platform crossings was lower ( Z=-2.673, P=0.008) in the sevoflurane group compared with those of the control group. (2) Results from Western blot and Golgi staining showed that the expression levels of hippocampal synapse-related proteins in the sevoflurane group, including PSD95 ((0.38±0.07), (1.00±0.21); t=4.885) and SYN1 ((0.30±0.10), (1.00±0.10); t=8.575), were lower than those in the control group, as well as the number of dendritic spines ((10.3±2.5), (20.0±1.0); t=6.183)(all P<0.05). (3) Transmission electron microscopy showed mitochondrial damage in the hippocampus of the sevoflurane group, and the expression level of ROS ((3.05±0.90), (0.97±0.16); t=-4.555, P=0.004) was significantly higher than that of the control group, while the expression level of GSH ((0.71±0.07), (1.00±0.09); t=5.396, P=0.002) was significantly lower than that of the control group.(4) Western blot and immunofluorescence demonstrated that hippocampal p-AMPK, p-GSK-3β (Ser9), and HO-1 expression levels in the sevoflurane group were significantly lower than those in the control group ( t=2.845, 7.087, 4.551, all P<0.05), and Nrf2 fluorescence intensity was also markedly reduced ( P<0.05). Conclusion:The cognitive impairment induced by sevoflurane in aged mice may be caused by mitochondrial damage, oxidative stress response and synaptic damage, which maybe associated with the inhibition of the AMPK/GSK-3β/Nrf2 signaling pathway.

Objective:To discuss the effect of sialic acid-binding immunoglobulin-like lectin-15(Siglec15)derived from M2 tumor-associated macrophages(M2-TAMs)on promoting the malignant biological behavior of the esophageal squamous cell carcinoma(ESCC)through bioinformatics analysis,and to validate the findings through cell experiment.Methods:The Tumor Immune Estimation Resource(TIMER)online Database was used to analyze the expression differences and immune infiltration of Siglec15 in pan-cancer and adjacent normal tissues.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Siglec15 mRNA in M2-TAMs and ESCC EC109 and KYSE150 cells.Based on the non-contact co-culture of M2-TAMs and ESCC cells,the following groups were set up,such as EC109/KYSE150 group,EC109/KYSE150+si-NC group(transfected with si-NC sequence),and EC109/KYSE150+si-Siglec15 group(transfected with si-Siglec15#1 and si-Siglec15#2 sequences).CCK-8 method was used to detect the proliferation activities of the cells in various groups;wound healing assay was used to detect the wound healing rates of the cells in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion cells in various groups;flow cytometry was used to detect the apoptotic rates of the cells in various groups.Results:The bioinformatics analysis results showed that compared with adjacent normal tissue,the expression levels of Siglec15 mRNA in pan-cancer tissues such as esophageal cancer,colon cancer,and head and neck squamous cell carcinoma tissues were increased(P＜0.05 or P＜0.01),and the expression level of Siglec15 mRNA in esophageal cancer tissue was significantly positively correlated with the infiltration of the macrophages(P＜0.05).Compared with the EC109 cells and KYSE150 cells,the expression level of Siglec15 mRNA in M2-TAMs was significantly increased(P＜0.01).There was no significant difference in the proliferation rate of the cells among EC109/KYSE150 group,EC109/KYSE150+si-NC group,and EC109/KYSE150+si-Siglec15 group(P＞0.05).Compared with EC109/KYSE150 group,after treated for 24 and 48 h,the wound healing rate of the cells in EC109/KYSE150+si-NC group was increased(P＜0.01),the numbers of migration and invasion cells were increased(P＜0.05),and the apoptotic rate was decreased(P＜0.01).Compared with EC109/KYSE150+si-NC group,the wound healing rates of the cells in EC109/KYSE150+si-Siglec15#1 group and EC109/KYSE150+si-Siglec15#2 group were decreased(P＜0.05),the numbers of migration and invasion cells were decreased(P＜0.05),and the apoptotic rates of the cells had no significant difference(P＞0.05).Conclusion:Siglec15 derived from M2-TAMs may be a key factor in promoting the migration and invasion of the ESCC cells.

Objective To investigate the expression,synergistic relationship and clinical significance of alcohol de-hydrogenase(ADH1A)and vascular endothelial growth factor-A(VEGFA)in hepatocellular carcinoma(HCC).Methods The expression and correlation of ADH1A and VEGFA in HCC and adjacent normal tissues were ana-lyzed by GEPIA.TCGA and GSEA were used to analyze the pathway of ADH1A in HCC.The clinical and patho-logical data of 84 patients with HCC were collected,and 54 patients with paracancer normal tissue samples were se-lected as controls to analyze the correlation between ADH1A and VEGFA and clinicopathological parameters of HCC.Immunohistochemistry was used to detect the protein expression of ADH1A and VEGFA in cases and con-trols,and the correlation between the expression of ADH1A and VEGFA and the clinical progression and prognosis of patients with HCC was analyzed based on clinical pathological parameters and Kaplan-Meier.Results Bioinfor-matics analysis found that ADH1A was low-expressed in HCC and VEGFA was highly expressed in HCC,and there was a negative correlation between the two(P<0.001);immunohistochemical detection results showed that the expression of ADH1A in HCC tissue was lower than that in normal tissue adjacent to cancer(P<0.01)while the expression rate of VEGFA in HCC tissue was significantly higher than that of normal tissue adjacent to cancer(P<0.01);The recurrence rate of vascular thrombus and HCC patients in HCC group with high expression of ADH1A was lower(P<0.05).The proportion of tumor diameter>5 cm,high TNM stage,microsatellite and G2-G3 dif-ferentiation in HCC tissues in VEGFA high expression group was higher(P<0.05).Kaplan-Meier survival analy-sis showed that patients with high ADH1A expression and low VEGFA expression had a higher five-year survival rate.Conclusion Low expression of ADH1A and high expression of VEGFA in tumor tissues of patients with HCC indicate tumor progression and can be used as one of the prognostic evaluation indicators for patients with HCC.

Background/Aims: The shortage of donor liver hinders the development of liver transplantation. This study aimed to clarify the poor outcomes of functionally marginal liver grafts (FMLs) and provide evidence for the improvement of ischemia-free liver transplantation (IFLT) after FML transplantation. Methods: Propensity score matching was used to control for confounding factors. The outcomes of the control group and FML group were compared to demonstrate the negative impact of FMLs on liver transplantation patients. We compared the clinical improvements of the different surgical types. To elucidate the underlying mechanism, we conducted bioinformatic analysis based on transcriptome and single-cell profiles. Results: FMLs had a significantly greater hazard ratio (HR: 1.969, P=0.018) than did other marginal livers. A worse 90-day survival (Mortality: 12.3% vs. 5.0%, P=0.007) was observed in patients who underwent FML transplantation. Patients who received FMLs had a significant improvement in overall survival after IFLT (Mortality: 10.4% vs 31.3%, P=0.006). Pyroptosis and inflammation were inhibited in patients who underwent IFLT. The infiltration of natural killer cells was lower in liver grafts from these patients. Bulk transcriptome profiles revealed a positive relationship between IL-32 and Caspase 1 (R=0.73, P=0.01) and between IL-32 and Gasdermin D (R=0.84, P=0.0012). Conclusions FML is a more important negative prognostic parameter than other marginal liver parameters. IFLT might ameliorate liver injury in FMLs by inhibiting the infiltration of NK cells, consequently leading to the abortion of IL-32, which drives pyroptosis in monocytes and macrophages.

OBJECTIVE To study pharmacodynamics and potential mechanism of Blumea balsamifera total flavonoids against acute myocardial infarction （AMI） model rats. METHODS AMI model of SD rats was established by ligating anterior descending branch of left coronary artery. Fifty model rats were randomly divided into model group （0.8% carboxymethyl cellulose solution）， positive control group （Compound danshen tablet， 300 mg/kg）， B. balsamifera total flavonoids low-dose， medium-dose and high- dose groups （3， 10， 30 mg/kg）， with 10 rats in each group. Other 10 rats were included in sham operation group （0.8% carboxymethyl cellulose solution）. After 1 day of surgery， they were given relevant medicine 3 mL/kg intragastrically， once a day， for 4 consecutive weeks. The changes of S-T segment were recorded before and after operation， after weekly intragastric administration. The hemodynamic indexes of rats were all determined， i.e. systolic blood pressure （SBP）， diastolic blood pressure （DBP）， mean arterial pressure （MAP）， left ventricular systolic pressure （LVSP）， left ventricular end diastolic blood pressure （LVEDP）， maximal left ventricular pressure rising rate （+LVdp/dtmax）， maximal left ventricular pressure decreasing rate （－LVdp/ dtmax）. The levels of serum myocardial enzymes [lactate dehydrogenase （LDH）， creatine kinase isoenzyme-MB （CK-MB）] and inflammatory factors [tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， IL-1β] were determined. The myocardial infarction rate of rats and the phosphorylation levels of phosphoinositide 3-kinase （PI3K） and protein kinase B （Akt） proteins in myocardial tissue were determined. RESULTS Compared with model group， S-T segments of electrocardiogram were all decreased significantly in administration groups （P＜0.05）. SBP， DBP， MAP， LVSP， +LVdp/dtmax， －LVdp/dtmax， and ratio of p-PI3KTyr607/ PI3K， p-AktThr308/Akt， p-Aktser473/Akt were increased significantly in B. balsamifera total flavonoids medium-dose and high-dose groups （P＜0.05）. The levels of LVEDP， serum myocardial enzymes and inflammatory factors， myocardial infarction rate were all decreased significantly （P＜0.05）. CONCLUSIONS balsamifera total flavonoids can improve cardiac function of AMI model rats， the mechanism of which may be associated with inhibiting the expression of inflammatory factor and activating PI3K/Akt signaling pathway.

Vagus nerve stimulation (VNS) is a neuromodulation technique that achieves therapeutic purpose through intermittent and chronic stimulation of vagus nerve afferent fibers, which has a remarkable effect on functional diseases of the central nervous system, and has been approved by FDA for intractable epilepsy, depression and migraine treatments. Transcutaneous auricular vagus nerve stimulation(taVNS) is a new type of non-invasive nerve regulation therapy based on traditional VNS and vagus nerve anatomy, and has a wide range of central regulation; taVNS can improve the cognitive state by regulating functions of cognition-related cerebral cortex and nerve nuclei, regulating inflammatory response, promoting neurotransmitter transmission, and so on, which has a broad application prospect in cognition-related diseases. This paper mainly summarizes the recent advance in central mechanism of taVNS in improving cognitive function.