Objective: To explore the correlation between Candida albicans and the development of oral leukoplakia (OLK), and to provide a basis for improving the pathogenic mechanism of the malignant transformation of OLK. Methods: Oral microbiome data were obtained from public databases (NCBI BioProject, PRJNA788378; GEO, GSE227919), and bioinformatic methods were employed to evaluate the correlation between Candida albicans infection and OLK. Approval was obtained from the institutional Medical Ethics Committee. A tissue microarray was constructed using samples collected from an OLK clinical cohort. Hematoxylin and eosin (H&E) staining and periodic acid-Schiff (PAS) staining were performed to analyze the relationship between the Candida albicans detection rate and clinicopathological features. Approval was obtained from the institutional Animal Ethics Committee. A mouse model was established by combining 4-nitroquinoline-1-oxide (4NQO) in drinking water with oral inoculation of Candida albicans (4NQO + Candida albicans group), while mice treated with 4NQO in drinking water and PBS served as the control group (4NQO + PBS group). The degree of epithelial dysplasia was compared between the two groups to assess the impact of Candida albicans infection on lesion progression (defined in this study as the progression from mild/moderate epithelial dysplasia to severe dysplasia/carcinoma in situ or invasive squamous cell carcinoma). Results: Bioinformatic analysis revealed that the detection rate of Candida albicans in OPMDs and OLK tissues was significantly higher than that in the healthy control group. Staining results of clinical samples demonstrated that Candida albicans colonized OLK lesions; compared with Candida albicans-negative patients, positive patients exhibited a state of high-grade progression. Animal experiments indicated that, compared with the 4NQO + PBS group, the degree of oral epithelial dysplasia in the 4NQO + Candida albicans group was significantly exacerbated, and the malignant transformation rate was higher, suggesting that Candida albicans promotes the high-grade progression of OLK. Conclusion Candida albicans exhibits a increasing trend during the malignant progression of the OLK. It aggravates the degree of epithelial dysplasia in OLK and promotes its transformation into high-grade lesions, suggesting that Candida albicans plays a crucial promoting role in the high-grade progression of OLK.

ObjectiveTo investigate whether Carthamus tinctorius L.-derived nanovesicles （CDNVs） can alleviate angiotensin Ⅱ （ANGⅡ）-induced cardiomyocyte （H9C2） hypertrophy by inhibiting the NF-κB signaling pathway and to elucidate the underlying molecular mechanisms. MethodsCDNVs were isolated and purified by ultracentrifugation combined with sucrose density gradient centrifugation， and their morphology and particle size distribution were identified by transmission electron microscopy and nanoparticle tracking analysis. Rat H9C2 cardiomyocytes were cultured in vitro and randomly divided into the normal control group （NC）， ANGⅡ （1.0 μmol/L） group， ANGⅡ+CDNVs group， and ANGⅡ+CDNVs+NF-κB activator 1 group. After 24 h of intervention， cell viability was detected by the CCK-8 assay， the expression of myocardial hypertrophy-related genes （ANP， BNP） was determined by qPCR， the relative cell area was measured by phalloidin staining， the expression of hypertrophy marker proteins and NF-κB pathway proteins （P-p65/p65， P-IκBα/IκBα） was examined by Western blot， and nuclear translocation of p65 was observed by immunofluorescence. Subsequently， NF-κB signaling pathway agonist activator 1 was added， and the expression of myocardial hypertrophy marker proteins and NF-κB pathway proteins was detected. ResultsCDNVs exhibited a typical round-like vesicle structure with an average particle size of 140.4 nm， which was consistent with the morphological characteristics of exosomes. ANGⅡ at concentrations of 0.1-1.0 μmol/L had no significant effect on cardiomyocyte viability. Compared with the NC group， ANGⅡ significantly upregulated the gene expression of ANP and BNP， increased the relative surface area of cardiomyocytes， elevated the levels of myocardial hypertrophy marker proteins and NF‑κB pathway proteins， and promoted the nuclear translocation of p65 （P<0.05）. Following treatment with CDNVs， all the above indicators were markedly reduced in comparison with the ANGⅡ group （all P<0.05）. However， all these experimental results were reversed after the addition of an NF‑κB agonist. ConclusionCDNVs alleviate ANGⅡ-induced H9C2 cell hypertrophy by inhibiting the NF-κB signaling pathway.

ObjectiveTo investigate the effect of growth hormone （GH） pretreatment on the clinical outcomes of in vitro fertilization/intracytoplasmic sperm injection-embryo transfer （IVF/ICSI-ET） in patients with diminished ovarian reserve （DOR）. MethodsA retrospective analysis was conducted on the clinical data of 359 DOR patients undergoing IVF/ICSI-ET. Based on the use of GH pretreatment before ovarian stimulation and/or its addition during the stimulation cycle， patients were divided into three groups： the Control group （n=119）， receiving no GH pretreatment or addition； the Non-GH pretreatment group （n=115）， receiving GH （4 IU/d） only during ovarian stimulation without pretreatment； and the GH pretreatment group （n=125）， pretreated with GH （2 IU/d） for 4-6 weeks before stimulation and also receiving GH （4 IU/d） during stimulation. The basic characteristics， ovarian stimulation parameters， embryological data， and cumulative live birth rate （CLBR） were compared among the groups. Results① No significant differences were found among the three groups regarding female age， male age， duration of infertility， body mass index （BMI）， basal hormone levels， distribution of ovarian stimulation protocols， endometrial thickness， serum estradiol （E2） and luteinizing hormone （LH） levels on the day of HCG trigger， or the number of blastocysts transferred per cycle. ② The GH pretreatment group required significantly lower total gonadotropin （Gn） dosage and fewer days of stimulation than the Control and Non-GH pretreatment group（P<0.05）. Compared to the Control group， the GH pretreatment group showed significantly higher numbers of retrieved oocytes， the number of mature oocytes， 2-pronuclei （2PN） zygotes， 2PN cleaved embryos， transferable embryos， high-quality embryos， as well as higher cumulative clinical pregnancy rates and CLBR （P<0.05）. Compared to the Non-GH pretreatment group， the GH pretreatment group had significantly more transferable embryos， high-quality embryos， and a higher CLBR （P<0.05）. ③ Compared to the Control group， the Non-GH pretreatment group only had a significantly higher number of retrieved oocytes （P<0.05）， with no significant differences in the number of mature oocytes， 2PN fertilized oocytes， 2PN cleaved embryos， transferable blastocysts， high-quality blastocysts， cumulative clinical pregnancy rate， and CLBR. ConclusionFor DOR patients， GH pretreatment for 4-6 weeks prior to ovarian stimulation can improve oocyte yield， embryo quality， cumulative clinical pregnancy rate， and CLBR， thereby enhancing the clinical outcomes of IVF/ICSI-ET.

OBJECTIVE To explore the mechanism of Yiqi yangyin huazhuo tongluo formula in ameliorating diabetic kidney disease （DKD） based on the gut microbiota-lipopolysaccharide （LPS）/Caspase-11/gasdermin D （GSDMD） pathway.METHODS A DKD mouse model was established by high-fat and high-sugar diet combined with intraperitoneal injection of low-dose streptozotocin. The successfully modeled mice were randomly divided into model group， metformin group （positive control， 200 mg/kg） and Yiqi yangyin huazhuo tongluo formula group （32.4 g/kg， calculated as crude drug dosage）， with 10 mice in each group； another 10 mice were set as the control group. Mice were intragastrically administered with corresponding drug solutions or normal saline once a day for 8 consecutive weeks. After the last administration， serum glycolipid metabolism indicators and urinary renal function indicators of mice were determined； the pathological and morphological changes of renal and colonic tissues were observed； LPS contents in renal tissues， colonic contents and serum were detected. 16S rRNA high-throughput sequencing was performed for gut microbiota analysis of colonic contents. The protein expressions of Occludin and zonula occludens-1 （ZO-1） in colonic tissues as well as Caspase-11 and GSDMD in renal tissues were detected.RESULTS Compared with the control group， the model group exhibited significantly elevated levels of serum fasting blood glucose， triglyceride， total cholesterol， serum creatinine， blood urea nitrogen and microalbumin， LPS contents in each sample， as well as protein expressions of Caspase-11 and GSDMD in renal tissues （ P ＜0.05）. The protein expressions of ZO-1 and Occludin in colonic tissues， together with Chao1 and Ace indices of gut microbiota， were markedly decreased （ P ＜0.05）， and the ratio of Firmicutes to Bacteroidetes was abnormally increased. Obvious histopathological changes were observed in renal and colonic tissues； the percentage of PAS-positive area in kidney was markedly increased （ P ＜0.05）. Compared with the model group， the Yiqi yangyin huazhuo tongluo formula group showed remarkable reductions in glycolipid metabolism and renal function-related indicators（except total cholesterol）， LPS contents in various samples， and renal protein expressions of Caspase-11 and GSDMD （ P ＜0.05）. The protein expression level of ZO-1 in colonic tissues and the Ace index of gut microbiota were significantly increased （ P ＜0.05）， and the relative abundance of beneficial genera including Bifidobacterium ， Faecalibaculum and Akkermansia in the gut microbiota was elevated.CONCLUSIONS The mechanism by which Yiqi yangyin huazhuo tongluo formula ameliorates DKD may be related to reshaping gut microbiota， repairing intestinal mucosal barrier， reducing LPS translocation into blood， and further inhibiting Caspase-11/GSDMD-mediated pyroptosis in kidney.

Objective:This study aims to investigate the differences in emotional status and cognitive function among patients with vestibular migraine (VM), Meniere′s disease (MD), and their comorbidity (VMMD), and to analyze key factors influencing cognitive function.Methods:This cross-sectional study included 96 outpatients (32 males, 64 females, aged 21-73 years) from the Department of Otorhinolaryngology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, between December 2022 and December 2023. The study population consisted of 31 VM patients (VM group), 36 MD patients (MD group), and 29 VMMD patients (VMMD group), along with 32 healthy controls (16 males, 16 females, aged 19-74 years). Cognitive function was assessed using the Montreal Cognitive Assessment (MoCA), while emotional status and somatization symptoms were evaluated through the Generalized Anxiety Disorder scale, Patient Health Questionnaire Depression scale, Symptom Checklist-90, and the Self-rating Somatization Symptom scale. Multiple linear regression analysis was conducted to explore the influence of different variables on cognitive function.Results:The total MoCA score in the VMMD group (26.0 [24.5, 28.0]) was significantly lower than that in the control group (28.0 [27.0, 29.0]) and the MD group (28.0 [26.0, 30.0]) ( P=0.006). VMMD patients exhibited significant impairments in specific cognitive domains, including visuospatial/executive function, delayed recall, and orientation ( P<0.05). Patients with VM, MD, and VMMD showed higher rates of anxiety, depression, and somatization symptoms compared to the control group ( P<0.05), with the VMMD group experiencing the most severe emotional distress. Multiple linear regression analysis identified education level and vestibular disease type as key factors affecting cognitive function, with a university-level education predicting higher MoCA scores ( P<0.001), while VMMD was associated with cognitive decline ( P<0.01). Conclusions:Patients with VM and MD, particularly those with comorbid VMMD, exhibit significant emotional distress. Cognitive impairments are present in VM and VMMD patients, affecting different cognitive domains. These factors should be comprehensively considered in clinical assessments to develop more effective treatment strategies.

Objective:To analyze the impact of changes in cardiovascular and cerebrovascular diseases mortality on the gender gap in life expectancy among Tianjin residents from 2004 to 2020.Methods:This is a cross-sectional study. The data sources included all-cause death surveillance data collected by the Tianjin Centre for Disease Control and Prevention, covering the entire population of Tianjin from 2004 to 2020, and the number of urban and rural residents by age, sex, and urban and rural areas from the Population Management Office of the Tianjin Public Security Bureau during the same period. The abbreviated life table and Arriaga′s decomposition method were applied to analyze the cause-of-death surveillance data of Tianjin residents from 2004 to 2020, and to calculate the impact of changes in mortality rates of cardiovascular and cerebrovascular diseases on gender differences in life expectancy by age, subcategory, and urban and rural areas.Results:In 2004, there were 56 189 deaths among Tianjin residents, including 31 233 males and 24 956 females. In 2020, there were 79 782 deaths among Tianjin residents, including 44 829 males and 34 953 females. The gender difference in life expectancy attributable to cardiovascular and cerebrovascular diseases among Tianjin residents increased from 1.52 years in 2004 to 2.02 years in 2020, with females having a longer life expectancy than males. The cumulative impact rate was higher among residents aged 40-74 years, increasing from 26.41% to 33.48%, while the cumulative impact rate among residents aged≥75 declined from 12.66% to 8.54%. The impact rates of intracerebral hemorrhage and cerebral infarction decreased significantly during the observation period, with intracerebral hemorrhage impact rate decreasing from 13.21% to 6.12% and cerebral infarction impact rate decreasing from 11.32% to 5.02%. The impact rate of acute myocardial infarction decreased but remained at a high level, from 9.70% to 6.99%. The impact rate of sequelae of cerebrovascular disease and other coronary heart diseases increased significantly, with the impact rate of sequelae of cerebrovascular disease increasing from 0.54% to 14.42% and the impact rate of other coronary heart diseases rising from 1.35% to 6.34%. The impact of sequelae of cerebrovascular disease deaths on the gender difference in life expectancy was greater in urban areas compared to rural areas, with an increase of 2 101.41% in urban areas and 1 898.51% in rural areas.Conclusions:The gender difference in life expectancy attributable to cardiovascular and cerebrovascular diseases among Tianjin residents is widening. Attention should be paid to the prevention of cerebrovascular disease sequelae and other coronary heart diseases among males aged of 40-74 years in urban areas.

Objective:To investigate the role of the Smad3 signaling pathway in the process of silica-induced epithelial-mesenchymal transition (EMT) .Methods:In September 2022, lung epithelial cells (BEAS-2B) were exposed to different concentrations of silica suspension (0, 50, 100, and 150 μg/ml) for 6 and 12 hours. Additionally, SIS3, a specific inhibitor of phosphorylated Smad3 (p-Smad3) , was utilized to establish the p-Smad3 inhibition model. The cells were divided into four groups: blank control gruop, silica group, SIS3 intervention group, and SIS3 +silica group. Cell morphology was observed using an inverted fluorescence microscope, while cell viability was assessed using a Cell Counting Kit-8 (CCK-8) . The mRNA and protein expression levels of E-cadherin (E-Cad) , N-cadherin (N-Cad) , Vimentin, Smad3, and p-Smad3 were analyzed by Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Differences between two groups were compared using Student's t-test, and multiple group comparisons were analyzed using a one-way analysis of variance with the Student-Newman-Keuls test.Results:Compared with the blank control group, the morphology of BEAS-2B cells shifted from epithelial to mesenchymal cell-like following silica exposure, and the cell viability of BEAS-2B cells declined after exposure to 150 μg/ml silica for 6 and 12 hours. Furthermore, silica exposure led to significant reductions in mRNA and protein expression levels of the epithelial cellular marker (E-Cad) in BEAS-2B cells, accompanied by increased expressions of interstitial cellular markers (N-Cad and Vimentin) . Importantly, the level of p-Smad3/Smad3 expression levels was also elevated in silica-treated cells ( P<0.05) . Compared to the blank control group, the level of p-Smad3/Smad3 expression levels was significantty reduced. Moreover, compared to the silica group, the protein expression levels of N-Cad and Vimentin in the cell of the SIS3+silica group were significantly reduced, while the E-Cad expression was increased ( P<0.05) . Conclusion:Silica exposure can prmote the epithelial mesenchymaol transformotion process by activating smod3 signa ling pathuay, and in hibiting smad3 signa ling pathuay can effctively alleviate the occurrence of epithelial mesenchymal transformation process.

Objective:To compare clinical features, esophageal motility, and reflux characteristics in elderly patients with gastroesophageal reflux disease(GERD)with and without extraesophageal symptoms.Methods:This study was conducted as a cross-sectional analysis involving 75 patients aged 60 years and older, who visited the outpatient department of gastroenterology at China-Japan Friendship Hospital between January and December 2023.Among these, 43 patients exhibited extraesophageal symptoms [23 males, 20 females; median age 69(66, 75)], while 32 patients did not present with such symptoms [12 males, 20 females; median age 69(66, 73.75)].We collected data regarding the patients' demographic characteristics, lifestyle habits, and comorbidities via a questionnaire, and analyzed the range of extraesophageal symptoms.The presence of reflux esophagitis was assessed according to the Los Angeles classification, and the status of the gastroesophageal valve was evaluated through gastroscopy.Additionally, we measured esophageal motility and reflux characteristics using high-resolution manometry and 24-hour pH-impedance monitoring.Results:The smoking rate was significantly higher in patients with extraesophageal symptoms compared to those without extraesophageal symptoms[39.53%(17/43) vs.15.63%(5/32), P=0.039].Conversely, the prevalence of hypertension was significantly higher in patients without extraesophageal symptoms compared to those with extraesophageal symptoms[41.86%(18/43) vs.65.63%(21/32), χ2=4.151, P=0.042].Among patients with extraesophageal symptoms, the proportion experiencing pharyngeal symptoms was the highest at 83.72% (36/43), followed by oral symptoms at 32.56%(14/43), while the lowest proportion was for respiratory symptoms at 16.28%(7/43).The esophageal body peristaltic contraction rate was significantly higher in patients without extraesophageal symptoms compared to those with extraesophageal symptoms[100%(100%, 100%) vs.100%(80%, 100%), Z=-2.671, P=0.008].The number of non-acid reflux episodes was significantly higher in patients with extraesophageal symptoms compared to those without[5.00(1.00, 5.00) vs.1.00(1.00, 2.00), Z=-3.842, P<0.001].Additionally, the proportion of proximal reflux was significantly higher in patients with extraesophageal symptoms compared to those without[83.8%(65.6%, 95.4%) vs.68.5%(59.8%, 77.35%), Z=-2.037, P=0.042]. Conclusions:Patients with GERD and extraesophageal symptoms exhibit a higher prevalence of smoking, diminished peristaltic function in the esophageal body, an increased incidence of proximal esophageal reflux, and a greater number of non-acid reflux episodes.

Objective:This study aimed to explore the molecular mechanisms of the maladaptation of H3N2 influenza virus in chicken embryos, provide a theoretical basis for the restoration of H3N2 influenza vaccine production in chicken embryos.Methods:Samples of respiratory secretions from patients with influenza-like symptoms (Influenza-like Illness, ILI) caused by H3N2 influenza virus were inoculated into chicken embryos and Madin-Darby Canine Kidney cells (MDCK), respectively. After isolating the virus, hemagglutination experiments were conducted to detect hemagglutination titers and hemagglutination inhibition experiments were used to compare antigenic differences; further, whole-genome sequencing of H3N2 influenza virus was performed using second-generation high-throughput gene sequencing (Next Generation High-Throughput Gene Sequencing, NGS), and key amino acid sites of mutations were identified through sequence alignment; combined with sialic acid receptor binding experiments, the differences in the binding of wild-type and mutant receptor binding sites (RBS) to sialic acid receptors were compared; finally, molecular docking and molecular dynamics simulation method were used to explore the specific molecular mechanisms of how mutation sites affect the differences in the affinity of the RBS pocket for sialic acid receptors.Results:The hemagglutination assay result indicated that both chicken embryos and MDCK cells could isolate the influenza virus, and the hemagglutination inhibition test showed that no antigenic differences were produced in the isolated strains. NGS analysis revealed that the H3N2 virus underwent an F195Y mutation in the (RBS) region of the hemagglutinin (HA) protein after adaptation through chicken embryo passages. Receptor-binding experiments demonstrated that the F195Y mutation enhanced the virus′s binding ability to α2, 3-linked sialic acid glycan (Neu5Acα2-3Galβ1-4GlcNAcβ-PAA, 3′SLN), while the mutation did not affect the affinity of the RBS pocket for α2, 6-linked sialic acid glycan (Neu5Acα2-6Galβ1-4GlcNAcβ-PAA, 6′SLN). Molecular docking and molecular dynamics simulation result indicate that the F195Y mutation, by replacing a hydrophobic amino acid with a hydrophilic one, leads to a significant decrease in the structure of the RBS pocket, enhancing the binding stability of the H3N2 influenza virus with α2, 3-sln. This is specifically manifested by an increase in binding time and an increase in the number of hydrogen bonds at the RBS site with the receptor. Furthermore, the F195Y mutation does not alter the binding of the virus to other receptors.Conclusions:The F195Y mutation in the RBS pocket of H3N2 influenza virus is a key site affecting the viral chicken embryo inadaptability.

Chronic hepatitis B（CHB）is one of the major challenges in the global public health field. As of 2022，approximately 254 million people worldwide were infected with the hepatitis B virus（HBV）. CHB is one of the main causes of liver cirrhosis and hepatocellular carcinoma（HCC）. Nucleos（t）ide analogs（NAs）and interferon therapy can delay the progression of liver fibrosis by inhibiting viral replication，but they cannot completely avoid the problem of heterogeneous treatment responses. Some patients are in a state of low-level viremia（LLV）during treatment. The persistent LLV state can induce chronic inflammation and the progression of liver fibrosis，ultimately increase the risk of HCC. In patients with poor treatment responses，the continuous active viral replication can induce immune disorders，accelerate the evolution of fibrosis to the decompensated stage of liver cirrhosis，and increase the risk of patient death. This article aims to review the definition，mechanisms，and impact on treatment outcomes of LLV and suboptimal response based on the latest research，provide a basis for optimizing antiviral therapy for CHB.