Objective: To explore new therapeutic strategies for periodontitis, this study prepared extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUC-MSCs) loaded with saikosaponin D (SSD) and evaluated their efficacy in a mouse model of periodontitis. Methods: This study was approved by the Medical Ethics Committee and the Laboratory Animal Ethics Committee of the unit. hUC-MSCs at logarithmic growth phase were seeded into 96 well plates and then treated with various concentrations of SSD for 24 hours. The CCK-8 assay was used to determine the SSD concentration that preserves hUC-MSC viability. Flow cytometry (FCM) assessed SSD-induced apoptosis in hUC-MSCs and determined drug loading and loading efficiency. Nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) characterized the morphology and size of hUC-MSCs-EVs and hUC-MSCs-SSD-EVs. C57BL/6J mice were divided into five groups with six mice per group: control group, periodontitis model group (PD), PD+SSD group (2 mg/kg), PD+hUC-MSCs-EVs group (10 μg/μL) and PD+hUC-MSCs-SSD-EVs group (10 μg/μL). Administration was performed 3 times a week for 4 consecutive weeks. Micro-CT and hematoxylin-eosin (HE) staining evaluated alveolar bone resorption in periodontal tissues of a mouse model of periodontitis. RT-qPCR measured inflammatory cytokine expression in periodontal tissues of a mouse model of periodontitis. Immunofluorescence analyzed macrophage M1/M2 polarization and runt-related transcription factor 2 (RUNX2) protein expression in periodontal tissues of a mouse model of periodontitis. Results: The CCK-8 assay showed that 10 μmol/L SSD had little effect on hUC-MSCs viability (P>0.05). FCM confirmed the successful loading of SSD into EVs with a loading efficiency of 11.40%. TEM and NTA revealed that hUC-MSCs-SSD-EVs presented round or oval membranous structures with diameters ranging from 200 to 300 nm. Micro CT and HE staining showed that, compared with the PD group, the distance from cementoenamel junction to alveolar bone crest (CEJ-ABC) was markedly shortened (P<0.01）.The alveolar bone of periodontitis‑model bone volume fraction (BV/TV) and trabecular number (Tb.N) were significantly elevated in the PD+hUC-MSCs-SSD-EVs group (P<0.01). RT-qPCR and IF results showed that hUC-MSCs-SSD-EVs in periodontal tissues markedly downregulated the expression of pro-inflammatory cytokines including interleukin-1β, interleukin-6, tumor necrosis factor-α, interleukin-12 and interleukin-22 (P<0.01), upregulated the expression of the anti-inflammatory factor interleukin-10 (P<0.01), promoted the polarization of macrophages from the M1 phenotype to the M2 phenotype, and elevated the protein expression level of RUNX2 (P<0.01). Conclusion hUC-MSC-SSD-EVs effectively alleviated inflammatory responses in a mouse model of maxillary periodontitis and reduced alveolar bone loss, providing a new idea for the treatment of periodontitis.

Alzheimer's disease(AD)is an age-related neurodegenerative disease with an increasing incidence worldwide.A large number of studies have shown that the incidence rates of hearing loss is high in patients with mild cognitive impairment and Alzheimer's disease,and may be a risk factor for the occurrence and development of cognitive impairment.There is an interaction between the two,but the causal mechanism is still unclear.Early screening and management of hearing impairment may play an important role in the early diagnosis,symptom im-provement and disease progression of Alzheimer's disease.This paper reviews relevant clinical and basic research to discuss the correlation between hearing loss and Alzheimer's disease,and the possible causal mechanism between them.

Objective:To explore the feasibility of active surveillance for multifocal papillary thyroid microcarcinoma(PTMC).Methods:It was a cross-sectional study. The thyroid ultrasonography data from 124 580 health check-up participants in the Health Management Center at the Affiliated Zhongshan Hospital of Dalian University between March 2017 and December 2023 were retrospectively analyzed. The patients were divided into unifocal or multifocal group according to the number of PTMC. The rates of growth, lymph node metastasis and progression in the unifocal and multifocal PTMC group during active surveillance were compared by using the Kaplan-Meier method and the log-rank test. The patients were divided into the group of progression or no-progression according to the outcome of active surveillance, and the basic clinical characteristics between the groups were compared. The Cox proportional hazards regression analysis was used to identify the risk factors for the progression of PTMC.Results:A total of 304 patients were enrolled in this study, among them, there were 239 cases of unifocal PTMC and 65 cases of multifocal PTMC. There was no significant differences in the rates of growth, lymph node metastasis and progression in the PTMC between the two groups during active surveillance (all P>0.05). During the active surveillance period, a total of 47 cases of PTMC progressed, while 257 cases did not. The progression rate was 15.5%. There was statistically significant difference in the initial age of PTMC diagnosis between the progression and non-progression group ( P<0.05). Multivariate regression analysis showed that age of initial diagnosis was the only risk factor for the progression of PTMC, the risk of progression decreased by 0.079 for every one-year increase in the initial diagnosis age [ HR=0.921, (95% CI: 0.888-0.955), P<0.001], multifocality was not a risk factor[ HR=1.973, 95% CI(0.972-4.462), P=0.103]. Conclusion:Active surveillance can be performed for multifocal PTMC in patients of appropriate age.

Alzheimer's disease(AD)is an age-related neurodegenerative disease with an increasing incidence worldwide.A large number of studies have shown that the incidence rates of hearing loss is high in patients with mild cognitive impairment and Alzheimer's disease,and may be a risk factor for the occurrence and development of cognitive impairment.There is an interaction between the two,but the causal mechanism is still unclear.Early screening and management of hearing impairment may play an important role in the early diagnosis,symptom im-provement and disease progression of Alzheimer's disease.This paper reviews relevant clinical and basic research to discuss the correlation between hearing loss and Alzheimer's disease,and the possible causal mechanism between them.

Objective:To explore the feasibility of active surveillance for multifocal papillary thyroid microcarcinoma(PTMC).Methods:It was a cross-sectional study. The thyroid ultrasonography data from 124 580 health check-up participants in the Health Management Center at the Affiliated Zhongshan Hospital of Dalian University between March 2017 and December 2023 were retrospectively analyzed. The patients were divided into unifocal or multifocal group according to the number of PTMC. The rates of growth, lymph node metastasis and progression in the unifocal and multifocal PTMC group during active surveillance were compared by using the Kaplan-Meier method and the log-rank test. The patients were divided into the group of progression or no-progression according to the outcome of active surveillance, and the basic clinical characteristics between the groups were compared. The Cox proportional hazards regression analysis was used to identify the risk factors for the progression of PTMC.Results:A total of 304 patients were enrolled in this study, among them, there were 239 cases of unifocal PTMC and 65 cases of multifocal PTMC. There was no significant differences in the rates of growth, lymph node metastasis and progression in the PTMC between the two groups during active surveillance (all P>0.05). During the active surveillance period, a total of 47 cases of PTMC progressed, while 257 cases did not. The progression rate was 15.5%. There was statistically significant difference in the initial age of PTMC diagnosis between the progression and non-progression group ( P<0.05). Multivariate regression analysis showed that age of initial diagnosis was the only risk factor for the progression of PTMC, the risk of progression decreased by 0.079 for every one-year increase in the initial diagnosis age [ HR=0.921, (95% CI: 0.888-0.955), P<0.001], multifocality was not a risk factor[ HR=1.973, 95% CI(0.972-4.462), P=0.103]. Conclusion:Active surveillance can be performed for multifocal PTMC in patients of appropriate age.

Objective:To explore the effectiveness and safety of percutaneous radiofrequency ablation for renal tumors, guided by both ultrasound and CT, under local anesthesia.Methods:A retrospective analysis was conducted on the clinical data of 40 patients with renal tumors admitted to Nanjing Drum Tower Hospital between January 2018 and December 2022. This treatment involved ultrasound/CT dual-guided radiofrequency ablation under local anesthesia. The cohort included 33 males and 7 females, with an average age of (61.5±11.9) years old and a body mass index (BMI) of (24.79±3.37) kg/m 2. The tumors were located in the left kidney in 20 cases and the right kidney in 16 cases, with 4 cases involving bilateral renal tumors. There were 44 tumors in 40 patients, with the maximum tumor diameter ranging from 1.0 to 4.0 cm [mean (2.3 ± 0.7) cm]. Distribution by kidney pole was as follows: 15 cases at the upper pole, 21 at the middle pole, and 8 at the lower pole. Of the tumors, 23 were exophytic, 5 were endophytic, and 16 exhibited mixed features. There were 2 patients with multiple metastases before surgery(including 1 patient with bilateral renal tumor). Preoperative serum creatinine level was 68.0(56.5, 87.5)μmol/L, and the estimated glomerular filtration rate (eGFR) was 114.2 (79.6, 132.4) ml/(min·1.73 m 2). All patients underwent renal biopsy before or during radiofrequency ablation. Percutaneous radiofrequency ablation surgery was performed using ultrasound and CT dual guidance on all patients, ensuring complete tumor destruction during the procedure as confirmed by dual positioning. Patients with bilateral tumors underwent two separate surgeries, spaced one month apart. Postoperatively, closely monitor the patient's vital signs and conduct long-term follow-ups to record any recurrence and metastasis. Results:In this series, all 40 procedures (involving 44 renal units) were successfully completed under local anesthesia without any need for blood transfusion, conversion to open surgery, or perioperative deaths. The average radiofrequency ablation time was (9.5 ± 3.6) min. Tumor characteristics included predominantly exophytic growths (23 cases, 52.3%), with 31 cases (70.5%) located more than 7 mm from the collecting system and 28 cases (63.6%) positioned posteriorly. Thirteen cases (29.5%) were entirely outside the polar line. The average R. E.N.A.L. nephrometry score was 6.1±0.2. Pathological examination revealed 34 cases of clear cell carcinoma, 2 of papillary renal cell carcinoma, 4 of unclassified renal cell carcinoma, and 4 benign renal tumors. In this cohort, two patients with bilateral renal tumors exhibited benign tumors on one side, while two other patients had malignant tumors in both kidneys. All 40 malignant tumors identified in 38 cases were classified at stage cT 1a.Postoperative serum creatinine level was 71.5 (59.0, 94.3) μmol/L, showing no statistically significant change from preoperative levels ( P > 0.05). Similarly, the eGFR post-operation was 107.4 (79.7, 132.2) ml/(min·1.73 m 2), which also did not differ significantly from preoperative values ( P > 0.05). There were no postoperative complications of Clavien-Dindo grade ≥Ⅱ, except for one case of severe pain (score 7-10). The follow-up period ranged from 15 to 70 months. Among the 38 cases, 36 patients did not have distant metastasis before surgery. There were 2 patients (5.5%) with local recurrence within 60 months after surgery. Among them, one case relapsed 6 months after radiofrequency ablation and was treated with partial nephrectomy. The patient was followed up for 60 months after the second treatment, and no local recurrence occurred. Another patient relapsed 41 months after surgery and was treated with radiofrequency ablation again. The patient was followed up for 12 months after the second treatment, and no local recurrence occurred. Two patients with distant metastasis before surgery were treated with targeted therapy plus immunotherapy for 12 months after surgery. One case had local recurrence 8 months after surgery and was treated with partial nephrectomy. The patient was followed up for 60 months after partial nephrectomy and no local recurrence occurred. Another patient with bilateral renal tumors developed left kidney recurrence 34 months after radiofrequency ablation, and underwent left partial nephrectomy. The right kidney recurred 42 months after radiofrequency ablation and underwent radiofrequency ablation again. After the second right renal radiofrequency ablation, no local recurrence occurred during 12 months of follow-up. Conclusions:Ultrasound/CT dual-guided percutaneous radiofrequency ablation, performed under local anesthesia for treating renal tumors, has minimal impact on the patient's renal function. The procedure boasts a low complication rate, with no postoperative severe complications. Additionally, the postoperative tumor control is effective, making it a safe and minimally invasive surgical option.

Objective:To investigate the factors related to the tumor growth in subjects with low-risk papillary thyroid microcarcinoma (PTMC) based on ultrasonography.Methods:This was a cross-sectional study. A total of 136 subjects who received health check-up in Health Management Center, the Affiliated Zhongshan Hospital of Dalian University from October 2017 to December 2023 were enrolled in the study. Low-risk PTMC were detected by ultrasonogrphy in those subjects and ultrashonography was followed up to observe the changes of maximum diameter and volume of the tumor, and metastasis of cervical lymph nodes. The clinical characteristics and ultrasonic image features were compared between the subjects with the tumor growth and without tumor growth, and the influencing factors of tumor growth were analyzed.Results:Among 136 subjects with low-risk PTMC, there were 23 cases (16.9%) with tumor growth (growth group) and 113 cases (83.1%) without tumor growth (non-growth group). Cervical lymph node metastasis occurred in 8 cases (5.9%: 7 (30.4%) in the growth group and 1 (0.9%) in non-growth group), no distant metastasis were detected. There were significantly differences in patients age of initial diagnoisi, maximum diameter and volume of tumors between the growth group and non-growth group (all P<0.05). Logistic regression analysis showed that age of initial diagnoisi ≤40 years ( OR=4.299, 95% CI:1.662-12.175, P=0.003) was an independent risk factor for tumor growth and the maximum diameter of the initial examination was independent protective factor for tumor growth (increasing 1 mm of initial diameter: OR=0.554, 95% CI:0.317-0.969, P=0.038). Conclusion:The size of most low-risk PTMC detected by ultrasonography during the health check-up does not grow and the risk of cervical lymph node metastasis is low; however, for those with age of initial diagnoisi ≤40 years and smaller size tumor, the risk of PTMC growth would be increased.

Objective To investigate the role of intestinal flora in the development of irritable bowel syndrome with diarrhea(IBS-D).Methods The mice treated with antibiotics were randomly assigned to experimental group or control group.IBS-D mouse feces and normal mouse feces were given to the experimental group and control group for fecal microbiota transplantation,respectively.The fecal water content,intestinal transit time,and intestinal sensitivity were observed.qRT-PCR was used to determine the mRNA expression of nuclear factor kappa-B(NF-κB),Toll like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),interleukin-1β(IL-Iβ),interleukin-6(IL-6),and tumor necrosis factor α(TNF-α)in colon tissue.Western blotting was used to determine the protein levels of the above-mentioned parameters.Results IBS-D related symptoms presented in the mice of the experimental group,such as increased fecal water content and intestinal sensitivity,and shortened intestinal transit time.The expression levels of inflammatory factors(IL-Iβ,IL-6,and TNF-α),NF-κB,TLR4,and MyD88 in the colon tissue of experimental group were significantly higher than those in the control group(P<0.05).Conclusion Intestinal flora inbalance is involved in the development and progression of IBS-D as an initiating important driver.Dysbiosis-induced hypo-inflammation of the intestinal mucosa and abnormal activation of NF-κB-related pathways may play key roles in this process.

Type Ⅶ secretion system(T7SS) is a novel and specialized secretion system discovered in recent years. It was first found in Mycobacterium tuberculosis. Type Ⅶ secretion system is involved in the secretion of virulence-associated proteins, the interaction between pathogens and hosts and the balance of zinc/iron ions. Moreover,it plays a critical role in the growth and pathogenesis of Mycobacteria. This review summarizes the components,substrates and translocation mechanisms of the type Ⅶ secretion system and its relation to the virulence of Mycobacteria aiming to provide references for developing novel strategies for disea-ses diagnosis,treatment and prevention.

The study of antigenic epitopes from Toxoplasma gondii has not only enhanced our understanding of the structure and function of antigens, the reactions between antigens and antibodies, and many other aspects of immunology, but it also plays a significant role in the development of new diagnostic reagents and vaccines. In the present study, T. gondii GRA6 epitopes were identified using bioinformatics tools and a synthetic peptide technique. The potential B cell epitopes of GRA6 predicted by bioinformatics tools concentrated upon 3 regions of GRA6, 1-20 aa, 44-103 aa, and 172-221 aa. Ten shorter peptides from the 3 regions were synthesized and assessed by ELISA using pig sera from different time points after infection. Three of the 10 peptides (amino acids 44-63, 172-191, and 192-211) tested were recognized by all sera and determined to be immunodominant B-cell epitopes of GRA6. The results indicated that we precisely and accurately located the T. gondii GRA6 epitopes using pig sera collected at different time points after infection. The identified epitopes may be very useful for further studies of epitope-based vaccines and diagnostic reagents.
Allergy and Immunology ; Antibodies ; Computational Biology ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; Epitopes, B-Lymphocyte* ; Indicators and Reagents ; Peptides ; Toxoplasma* ; Vaccines