Objective:To investigate the genetic mutation characteristics of patients with bilateral macronodular adrenocortical disease（BmacAD）and to analyze the clinical phenotypes and adrenal nodule transcriptomic profiles associated with different genotypes.Methods:This retrospective study analyzed the clinical data of 37 patients diagnosed with BmacAD by clinical and pathological assessment who underwent surgery at Peking Union Medical College Hospital from January 2001 to December 2024. Data included symptoms and signs related to hypercortisolism，24-hour urinary free cortisol，plasma adrenocorticotropic hormone（ACTH），and overnight low-dose dexamethasone suppression test results. Whole-exome sequencing（WES）was performed on frozen adrenal hyperplasia tissue samples from surgical specimens and matched frozen residual blood samples for all patients to screen for germline and somatic mutations. Sanger sequencing was used to validate some of the identified mutations. The relation between genetic mutations and clinical features was analyzed. Transcriptome sequencing of the hyperplastic tissues was conducted，and unsupervised clustering，differential gene expression analysis，and pathway enrichment analysis were used to compare the transcriptomic profiles of patients with different genotypes.Results:A total of 37 cases were included（23 males and 14 females），aged between 31?68 years old. Among the patients with available clinical data，most exhibited signs of hypercortisolism，including hypertension（100.0%，36/36），diabetes（50.0%，18/36），and centripetal obesity（55.9%，19/34），accompanied by relevant endocrine abnormalities such as elevated 24-hour urinary free cortisol（79.4%，27/34），suppressed ACTH（77.8%，28/36），and a positive overnight low-dose dexamethasone suppression test（45.2%，14/31）. WES and Sanger sequencing revealed that ARMC5 gene mutations were the most common. Among the 37 patients，33（89.2%）had ARMC5 germline mutations，and 31（83.8%）concurrently harbored ARMC5 somatic mutations，including 3 cases of loss of heterozygosity. Two patients without detectable ARMC5 mutations both carried a somatic p.S45P mutation in the CTNNB1 gene and neither had typical manifestations of hypercortisolism. Transcriptome sequencing results suggested that patients with ARMC5 mutations had an upregulation of pathways related to steroid secretion.Conclusions:The majority of BmacAD patients present with typical manifestations of hypercortisolism，while a minority exhibit only mild autonomous cortisol secretion without obvious symptoms. Biallelic inactivation of the ARMC5 gene is the primary genetic driver of this disease，leading to more severe hypercortisolism by upregulating steroid hormone secretion. CTNNB1 mutations may be involved in the pathogenesis of some patients without ARMC5 mutations.

Objective:To evaluate the efficacy and safety of immunotherapy combined with chemotherapy as conversion therapy for initially unresectable locally advanced esophageal squamous cell carcinoma.Methods:This retrospective case series study analyzed clinical and pathological data of 32 patients with initially unresectable locally advanced esophageal squamous cell carcinoma who received immunotherapy combined with chemotherapy at the Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University, from June 2020 to December 2024. The cohort included 27 males and 5 females, with an age ( M(IQR)) of 61(9)years (range:46 to 73 years). Five patients were diagnosed with stage Ⅲ, 27 with stage ⅣA. All patients received PD-1 inhibitor sintilimab combined with nedaplatin and albumin-bound paclitaxel. Radiological evaluations were performed every two cycles, the multidisciplinary team evaluation was conducted to determine conversion to resectable status, and patients with successful conversion underwent radical esophagectomy. Follow-up was conducted via telephone or outpatient visits every 3 to 6 months after the last treatment. The primary endpoint was R0 resection rate, secondary endpoints included objective response rate (ORR), pathological complete response (pCR) rate, major pathological response (MPR) rate, event-free survival (EFS), disease-free survival (DFS) in patients with R0 resection, overall survival (OS) and safety. Kaplan-Meier method was used to plot survival curves and estimate median EFS, DFS, OS rates and their 95% CI. The 95% CI for ORR, pCR rate, MPR rate, and downstaging rate were calculated using the Clopper-Pearson method. Results:The median treatment cycle of 2(1) (range:2 to 8). As of June 2025, the median follow-up was 32.5(13.5)months (range:6.4 to 59.1 months). Among the 32 patients, 9 experienced progression or recurrence, including 2 with liver and lymph node metastases, 2 with lung metastases, 2 with thoracic vertebral metastases, and 3 with mediastinal lymph node metastases. After conversion therapy, 29 patients underwent surgery, achieving an R0 resection rate of 84.4% (95% CI:67.2% to 94.7%), a pCR rate of 27.6% (95% CI:12.7% to 47.2%), and an MPR rate of 55.2% (95% CI:35.7% to 73.6%). Grade 3 or higher surgical complications occurred in 6.9%(2/29) of patients, and grade 3 or higher treatment-related adverse events were observed in 15.6%(5/29). Among the 32 patients, the ORR was 56.3% (95% CI:37.7% to 73.6%),the 3-year EFS rate and OS rate was 59.4% (95% CI:40.8% to 86.4%) and 59.7% (95% CI:40.0% to 89.0%) respectively. Conclusion:Immunotherapy combined with chemotherapy demonstrates high conversion rates and favorable safety in the conversion therapy of initially unresectable locally advanced esophageal squamous cell carcinoma, representing a promising treatment strategy.

Objective:To investigate the genetic mutation characteristics of patients with bilateral macronodular adrenocortical disease（BmacAD）and to analyze the clinical phenotypes and adrenal nodule transcriptomic profiles associated with different genotypes.Methods:This retrospective study analyzed the clinical data of 37 patients diagnosed with BmacAD by clinical and pathological assessment who underwent surgery at Peking Union Medical College Hospital from January 2001 to December 2024. Data included symptoms and signs related to hypercortisolism，24-hour urinary free cortisol，plasma adrenocorticotropic hormone（ACTH），and overnight low-dose dexamethasone suppression test results. Whole-exome sequencing（WES）was performed on frozen adrenal hyperplasia tissue samples from surgical specimens and matched frozen residual blood samples for all patients to screen for germline and somatic mutations. Sanger sequencing was used to validate some of the identified mutations. The relation between genetic mutations and clinical features was analyzed. Transcriptome sequencing of the hyperplastic tissues was conducted，and unsupervised clustering，differential gene expression analysis，and pathway enrichment analysis were used to compare the transcriptomic profiles of patients with different genotypes.Results:A total of 37 cases were included（23 males and 14 females），aged between 31?68 years old. Among the patients with available clinical data，most exhibited signs of hypercortisolism，including hypertension（100.0%，36/36），diabetes（50.0%，18/36），and centripetal obesity（55.9%，19/34），accompanied by relevant endocrine abnormalities such as elevated 24-hour urinary free cortisol（79.4%，27/34），suppressed ACTH（77.8%，28/36），and a positive overnight low-dose dexamethasone suppression test（45.2%，14/31）. WES and Sanger sequencing revealed that ARMC5 gene mutations were the most common. Among the 37 patients，33（89.2%）had ARMC5 germline mutations，and 31（83.8%）concurrently harbored ARMC5 somatic mutations，including 3 cases of loss of heterozygosity. Two patients without detectable ARMC5 mutations both carried a somatic p.S45P mutation in the CTNNB1 gene and neither had typical manifestations of hypercortisolism. Transcriptome sequencing results suggested that patients with ARMC5 mutations had an upregulation of pathways related to steroid secretion.Conclusions:The majority of BmacAD patients present with typical manifestations of hypercortisolism，while a minority exhibit only mild autonomous cortisol secretion without obvious symptoms. Biallelic inactivation of the ARMC5 gene is the primary genetic driver of this disease，leading to more severe hypercortisolism by upregulating steroid hormone secretion. CTNNB1 mutations may be involved in the pathogenesis of some patients without ARMC5 mutations.

Objective:To evaluate the efficacy and safety of immunotherapy combined with chemotherapy as conversion therapy for initially unresectable locally advanced esophageal squamous cell carcinoma.Methods:This retrospective case series study analyzed clinical and pathological data of 32 patients with initially unresectable locally advanced esophageal squamous cell carcinoma who received immunotherapy combined with chemotherapy at the Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University, from June 2020 to December 2024. The cohort included 27 males and 5 females, with an age ( M(IQR)) of 61(9)years (range:46 to 73 years). Five patients were diagnosed with stage Ⅲ, 27 with stage ⅣA. All patients received PD-1 inhibitor sintilimab combined with nedaplatin and albumin-bound paclitaxel. Radiological evaluations were performed every two cycles, the multidisciplinary team evaluation was conducted to determine conversion to resectable status, and patients with successful conversion underwent radical esophagectomy. Follow-up was conducted via telephone or outpatient visits every 3 to 6 months after the last treatment. The primary endpoint was R0 resection rate, secondary endpoints included objective response rate (ORR), pathological complete response (pCR) rate, major pathological response (MPR) rate, event-free survival (EFS), disease-free survival (DFS) in patients with R0 resection, overall survival (OS) and safety. Kaplan-Meier method was used to plot survival curves and estimate median EFS, DFS, OS rates and their 95% CI. The 95% CI for ORR, pCR rate, MPR rate, and downstaging rate were calculated using the Clopper-Pearson method. Results:The median treatment cycle of 2(1) (range:2 to 8). As of June 2025, the median follow-up was 32.5(13.5)months (range:6.4 to 59.1 months). Among the 32 patients, 9 experienced progression or recurrence, including 2 with liver and lymph node metastases, 2 with lung metastases, 2 with thoracic vertebral metastases, and 3 with mediastinal lymph node metastases. After conversion therapy, 29 patients underwent surgery, achieving an R0 resection rate of 84.4% (95% CI:67.2% to 94.7%), a pCR rate of 27.6% (95% CI:12.7% to 47.2%), and an MPR rate of 55.2% (95% CI:35.7% to 73.6%). Grade 3 or higher surgical complications occurred in 6.9%(2/29) of patients, and grade 3 or higher treatment-related adverse events were observed in 15.6%(5/29). Among the 32 patients, the ORR was 56.3% (95% CI:37.7% to 73.6%),the 3-year EFS rate and OS rate was 59.4% (95% CI:40.8% to 86.4%) and 59.7% (95% CI:40.0% to 89.0%) respectively. Conclusion:Immunotherapy combined with chemotherapy demonstrates high conversion rates and favorable safety in the conversion therapy of initially unresectable locally advanced esophageal squamous cell carcinoma, representing a promising treatment strategy.

OBJECTIVE To investigate the effects of soybean isoflavones （SI） on the reproductive development of young mice. METHODS C57BL/6 young mice were randomly divided into control group， SI low-dose and high-dose groups （10， 100 mg/kg）， with 10 mice in each group （half male and half female）. The young mice in each group were given corresponding liquid intragastrically， once a day， for 2 consecutive weeks. After the last administration， the percentage of body weight increase was calculated； serum estradiol and testosterone levels， malondialdehyde （MDA） content， total antioxidant capacity （T-AOC）， superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） activities in the reproductive organs of the young mice were determined. The histopathological changes in the reproductive organs were observed. The cell apoptosis of reproductive organs was detected. RESULTS Compared with the control group， the percentage of body weight increase in female mice was increased significantly in the SI high-dose group， while that of male mice was decreased significantly （P＜0.05 or P＜0.01）. Cystic follicles could be seen in the ovarian tissue in SI groups， a loose arrangement of spermatocytes could be seen in the testicular tissue， and partial epithelial cell shedding could be seen in epididymal tissue. The serum level of testosterone in female young mice and the serum levels of testosterone and estradiol in male young mice in SI groups， GSH-Px activity in the ovarian tissue of female young mice in the SI low-dose group， T-AOC activities in the ovarian tissue of female young mice in SI groups as well as the apoptotic rates of cells in testicular and epididymal tissue of male young mice in SI groups were increased significantly （P＜0.05 or P＜ 0.01）； the serum level of estradiol in female young mice in SI groups， SOD activity in the ovarian tissue of female young mice in the SI high-dose group， and MDA contents in the ovarian tissue of female young mice in SI groups as well as the apoptotic rates of cells in ovarian tissue of female mice in SI groups were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS SI can enhance the antioxidant stress capacity of ovarian tissue in female young mice and reduce their oxidative stress damage， but it has certain toxicity to reproductive organs in male mice.

Objective:To analyze retinal vascular parameters and distribution characteristics in Chinese population via the fully automated quantitative measurement of retinal vascular morphological parameters based on artificial intelligence technology.Methods:A cross-sectional study was performed.A total of 1 842 patients without fundus diseases who visited Beijing Tongren Hospital from January 2011 to December 2021 were included.Standardized questionnaires, blood draws and ophthalmologic examinations of enrolled subjects were conducted.Color fundus photographs centered on the optic disk of one eye of patients were collected, and a deep learning-based semantic segmentation network ResNet101-Unet was used to construct a vascular segmentation model for fully automated quantitative measurement of retinal vascular parameters.The main measurement indexes included retinal vascular branching angle, vascular fractal dimension, average vascular caliber, and average vascular tortuosity.To compare different retinal parameters between sexes, the correlation between the above parameters and ocular factors such as best corrected visual acuity, intraocular pressure, and axial length, as well as systemic factors such as sex, age, hypertension, diabetes mellitus, and cardiovascular disease was analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (No.20001220). Written informed consent was obtained from each subject.Results:The model established in this study achieved an accuracy over 0.95 for both vascular and optic disk segmentation.The vascular branching angle, vascular fractal dimension, average vascular caliber, and average vascular tortuosity were (51.023±11.623)°, 1.573(1.542, 1.592), 64.124(60.814, 69.053)μm, (0.001 062±0.000 165)°, respectively.Compared with females, males had larger vascular branching angle, smaller average vascular caliber and smaller vascular tortuosity, and the differences were statistically significant (all at P<0.05). The average vascular caliber increased by 1.142 μm in people with cardiovascular disease compared to people without cardiovascular disease ( B=1.142, P=0.029, 95% CI: 0.116-2.167). The average vascular tortuosity was positively correlated with hypertension ( B=3.053×10 -5, P=0.002, 95% CI: 1.167×10 -5-4.934×10 -5) and alcohol consumption ( B=1.036×10 -5, P=0.014, 95% CI: 0.211×10 -5-1.860×10 -5) and negatively correlated with hyperlipidemia ( B=-2.422×10 -5, P=0.015, 95% CI: -4.382×10 -5-0.462×10 -5). For each 1-mm increase in axial length, there was a decrease of 0.004 in vessel fractal dimension ( B=-0.004, P<0.001, 95% CI: -0.006--0.002), a decrease of 0.266 μm in the average vessel caliber ( B=-0.266, P=0.037, 95% CI: -0.516--0.016), and a decrease of -2.45×10 -5° in the average vessel tortuosity ( B=-2.45×10 -5, P<0.001, 95% CI: -0.313×10 -5--0.177×10 -5). For each 1.0 increase in BCVA, there was an increase of 3.992° in the vascular branch angle ( B=3.992, P=0.004, 95% CI: 1.283-6.702), an increase of 0.090 in vascular fractal dimension ( B=0.090, P<0.001, 95% CI: 0.078-0.102) and a decrease of 14.813 μm in the average vascular diameter ( B=-14.813, P<0.001, 95% CI: -16.474--13.153). Conclusions:A model for retinal vascular segmentation is successfully constructed.Retinal vessel parameters are associated with sex, age, systemic diseases, and ocular factors.

Objective:To investigate the differential expression of microRNA (miR)-214-3p in plasma exosomes in different types of uveal melanoma (UM) and evaluate whether miR-214-3p is a potential molecular biomarker for the diagnosis and prognosis of UM.Methods:Twenty-five UM in situ patients who received the enucleation of eyeball were enrolled at Beijing Tongren Hospital from December 2015 to October 2019, including 10 with epithelioid cell melanoma and 10 with spindle cell melanoma as well as 5 metastatic UM patients (1 with spindle cell-like melanoma and 4 with epithelioid cell-like melanoma) and 10 healthy subjects were enrolled during the same period.Blood sample was collected from all the subjects for the isolation of plasma exosomes.The morphology of exosomes was examined under the electron microscope.The exsomal marker proteins were identified by Western blot.The expression level of miR-214-3p in plasma exosomes was detected by real-time fluorescence quantitative PCR.The differential expression of miR-214-3p among different types of UM patients and healthy controls was compared.The diagnostic and classification performance of exosomal miR-214-3p was evaluated using receiver operating characteristic curve.Written informed consent was obtained from each subject prior to entering the study cohort.This study protocol was approved by an Ethics Committee of Capital Medical Univeristy (No.TRECKY2018-056).Results:The isolated exosomes were hemispherical in shape with a concavity on one side.The diameter of the exosomes was about 100 nm and the particle diameter of vesicles from samples was (82.0±2.7) nm.TSG101 protein was detectable and Calnexin protein was not found in the exosomes.The relative expression levels of plasma exosomal miR-214-3p in healthy control group, in situ UM group and metastatic UM group were 0.86(0.57, 1.49), 0.24(0.10, 0.67), and 0.43(0.23, 0.56), respectively.The miR-214-3p relative expression level in plasma exosomes of in situ UM patients and metastatic UM patients was significantly lower than that of healthy controls, and the differences were statistically significant ( Z=2.62, P<0.01; Z=2.08, P<0.05). The relative expression levels of exosomal miR-214-3p in spindle cell-like UM group and epithelioid cell-like UM group were 0.11(0.07, 0.64) and 0.46(0.14, 0.91), respectively, and no significant difference was found in the expression level of plasma exosomal miR-214-3p among different types of UM (all at P>0.05). The area under the curve of plasma exosomal miR-214-3p for UM diagnosis was 0.795. Conclusions:Plasma exosomal miR-214-3p level is significantly reduced in both in situ UM patients and metastatic UM patients.Plasma exosomal miR-214-3p is a new potential diagnostic biomarker for UM, but the exosomal miR-214-3p appears to not be able to distinguish the types of UM.

Objective To investigate the characteristics of immunophenotyping and clinical significance of MRD analysis in MM patients. Methods Multi-parameter flow cytometry was applied to analyze the immunophenotyping of malignant plasma cells from 172 MM patients, and normal plasma cells from 16 healthy individuals. MRD was analyzed in 32 MM patients with remission. Meanwhile, the effects of MRD status on the disease relapse and patient disease free survival ( DFS ) time was evaluated by following up patients. Results The immunophenotyping of normal plasma cells were CD38, CD138, CD19 and CD45 positive, while the predominant phenotype of MM cells were CD+38( 100.0% ), CD+138( 100.0% ), CD-19 ( 167/172, 97. 1% ), CD+56( 152/172, 88.4% ) and CD-45( 166/172, 96. 5% ). The characteristic markers for MM cells were CD+38, CD-138, CD-19, CD-45 and CD+56. MRD analysis revealed that, among 32 MM patients with remission, 14 patients were MRD negative and 18 patients were MRD positive. During follow-up of 2 to 16 months, the relapse rate in MRD negative patients was significantly lower ( 4/14, 28.6% ) than that of MRD positive patients ( 13/18, 72. 2% ;χ2 =6. 03, P ＜0. 05 ). Furthermore, the DFS time was significantly longer in MRD negative patients[ 16. 23( 10. 37-21.62 )months ] than that of the MRD positive patients [ 10. 07( 3. 79-16. 20 )months,χ2 =7. 53,P ＜0. 05 ]. Conclusions CD+38, CD+138, CD-19, CD-45 and CD+56 are the characteristic markers of MM cells compared to those of the normal plasma cells. MRD analysis is a valuable prognostic factor for MM patients.

Objective To investigate expressions of CDev6 and p-ERK1/2 in AML patients and its clinical significance.Methods Expressions of CD44v6 and p-ERK1/2 on bone marrow blasts in 152 AML patients and 22 normal controls were determined by flow cytometry.Meanwhile,the effects of CD44v6 expression(CD+44v6 and CD-44v6) on relapse rate and survival time were analyzed by following up of 129 AML patients.Results Double positive expressions of CD44v6 and p-ERK1/2 were observed in 4.5%(1/22) of the normal bone marrow blasts while single positive expression of CD44v6 and p-ERK1/2 was observed in 36.2%(55/162) and 64.5%(97/162) of AML patients,respectively.The MFI of p-ERK1/2 expression on blast cells in AML patients was [14(7 -58)],which was higher than that in normal controls [8(6 - 10),U =4.2,P＜0.01].Furthermore,MFI of p-ERK1/2 expression on blast cells in CD+44v6 AML patients was [28(15-61)] ,which was significantly higher than that in CD-44v6 AML patients [18(6- 37),U =6.7,P＜0.01].A strong correlation was obtained between CD44v6 and p-ERK1/2 expression(rs = 0.7,P＜ 0.01).Among 129 patients followed up for 4 to 65 months,the data also revealed that the relapse rate of CD-44v6 AML patients was 32.1%(26/81) ,which was much lower than that in CD44v6 AML patients [81.3%(39/48),x2 = 29.13,P＜ 0.01).And the overall survival in CD44v6 AML patients was(28.5 ± 1.8)months,which was significantly worse than that of CD44v6 AML patients [(51.2±2.0) months,x2 =48.2,P＜ 0.01).Conclusion CD44v6 expression was associated with a poor survival in AML patients,and CD44v6 might promote the expansion of leukemic blast cells by up-regulating p-ERK1/2.

In order to know that Streptococcus exist in health swine herd,2 204 cases of nose swabs were collected in swine from different areas in Dongbei three province of northeast China.Seed swab to Streptococcus culture medium,24 hours after culturing,gram staining with becteria culture,gram positive Streptococcus were identified with PCR technology,then farther identified serotype with Streptococcus suis 1,2,7,9 serotype special primers.The results showed that Streptococcus exist rate in swine herd of Heilongjiang,Jinlin,Liaoning is 29 %,27 %,34 %,respectively.PCR typing assay indicated that 155 strains of Streptococcus suis were isolated including 7 strains of 1 serotype,39 strains of 2 serotype,4 strains of 7 serotype,11 strains of 9 serotype,94 strains of other serotype in three province of northeast China.