OBJECTIVE: To explore the serological and molecular genetic characteristics of a family with subtype B(A)06. METHODS: A neonatal hyperbilirubinemia patient who was treated at Henan Children's Hospital on June 15, 2023 due to "yellowing of the skin and gradual aggravation", and was found to have inconsistent ABO forward and reverse typing through blood type testing, was selected as the research subject. Six milliliters of peripheral blood were collected from the newborn and her family members (grandfather, grandmother, father, mother and aunt) respectively. ABO blood group identification was performed by the blood group serological method. Human genomic DNA was extracted using the nucleic acid extraction or purification reagent BT-01. ABO gene exons 2 to 7 were amplified by PCR. The PCR-specific products that were successfully amplified were sequenced by Sanger method. Taking ABO*A1.01 as the reference sequence, the ABO gene sequences of the newborn and her family members were analyzed to determine the ABO genotype. The procedures followed in this study were approved by the Ethics Committee of Henan Children's Hospital (Ethics No.: 2022-K-L036). RESULTS: The serological results of ABO blood group showed that the newborn, her grandfather, father and aunt were all incompatible with the forward and reverse typing. The blood group phenotype of the newborn was AwB or B(A), the blood group phenotype of the grandfather was A2B or B(A), the blood group phenotype of the father and aunt were A2B, and the blood group phenotype of the grandmother and mother were both O. The screening test results of hemolytic disease of the newborn showed that the free test detected IgG anti-A1 antibody, while the elution test, direct antiglobulin test and antibody screening results were all negative. The Sanger sequencing results showed that the newborn had variations of c.261delG, c.297A>G, c.526C>G, c.657C>T, c.703G>A, c.796C>A and c.930G>A. Her grandfather had variations of c.297A>G, C.526C>G, c.657C>T, c.703G>A, c.796C>A, c.803G>C and c.930G>A. Her grandmother had variations of c.106G>T, c.188G>A, c.189C>T, c.220C>T, c.261delG, c.297A>G, c.646T>A, c.681G>A, c.771C>T and c.829G>A. Her father and aunt had variations of c.106G>T, c.188G>A, c.189C>T, c.220C>T, c.261delG, c.297A>G, c.526C>G, c.646T>A, c.657C>T, c.681G>A, c.703G>A, c.771C>T, c.796C>A, c.829G>A and c.930G>A. Her mother had variations of c.106G>T, c.188G>A, c.189C>T, c.220C>T, c.261delG, c.297A>G, c.646T>A, c.681G>A, c.771C>T, and c.829G>A.The genotype of the newborn was ABO*BA.06/ABO*O.01.01, her grandfather was ABO*BA.06/ABO*B.01, her grandmother was ABO*O.01.02/ABO*O.01.02, her father and aunt were ABO*BA.06/ABO*O.01.02, and her mother was ABO*O.01.01/ABO*O.01.02. The ABO*BA.06 allele of the newborn, grandfather, father and aunt was caused by the c.803C>G variation in exon 7 based on the ABO*B.01 allele. The ABO*BA.06 allele can be stably inherited in this family. CONCLUSION The blood type of neonatal patients with B(A)06 subtype can be accurately determined by gene sequencing technology. If the forward typing is ≤ 3+ agglutination intensity in newborn ABO blood group identification, the reason should be carefully analyzed, and the molecular biology technology and family gene sequencing results should be used to jointly determine if necessary.
Humans ; ABO Blood-Group System/genetics* ; Female ; Pedigree ; Male ; Infant, Newborn ; Asian People/genetics* ; Genotype ; China ; Blood Grouping and Crossmatching ; Hyperbilirubinemia, Neonatal/blood* ; East Asian People

Objective To investigate the clinicopathological characteristics, diagnostic key points, and treatment strategies of penile metastatic extramedullary plasmacytoma(EMP). Case Report A 47-year-old male patient presented with a painless indurated nodule on the dorsal side of the penis and had a previous history of plasmacytoma in the nasal cavity and tibia. He subsequently underwent complete excision of the penile lesion. Postoperative histopathological examination confirmed the diagnosis of anaplastic plasmacytoma. Immunohistochemical analysis showed positive staining of CD138, CD38, MUM1, and CD79a, with Ki-67(+60%). Light chain restriction was also identified. No local recurrence or distant metastasis was observed during the 5-month follow-up. Conclusion Metastatic EMP of the penis is an extremely rare condition, and the diagnosis relies on pathological examinations combined with immunohistochemical analysis. Surgical resection can be an effective treatment modality for localized lesions;however, long-term follow-up is essential to monitor disease progression.

Extracellular vesicles(EVs)are nanoscale membrane vesicles released by cells that carry bioactive molecules such as proteins and nucleic acids. They play a key role in intercellular communication. In recent years, they have shown significant potential in the diagnosis and treatment of corneal and ocular surface diseases. As diagnostic biomarkers, EVs derived from tears or other sources can reflect disease-specific characteristics, providing a basis for the early disease detection. In terms of therapy, mesenchymal stem cells(MSCs)-derived EVs deliver miRNAs to modulate inflammatory pathways, promote corneal epithelial repair, and inhibit fibrosis. Furthermore, as natural drug vehicles, EVs enable precise delivery of therapeutic agents and promote corneal wound healing. This review systematically summarizes the recent research advances in the application of EVs in the field.

Objective:To investigate the clinicopathological characteristics and prognostic factors of biliary neuroendocrine neoplasms (NENs).Methods:The retrospective cohort study was conducted. The clinicopathological data of 36 patients who underwent surgical treatment for biliary NENs at The First Hospital of Jilin University from January 2013 to December 2023 were collected. There were 22 males and 14 females, aged (59±9)years. Observation indicators: (1) clinicopatholo-gical characteristics of patients; (2) follow-up; (3) prognostic factors analysis of patients. Compari-son of measurement data with normal distribution among multiple groups was conducted using the ANOVA. Comparison of measurement data with skewed distribution among multiple groups was conducted using the Kruskal-Wallis H test. Comparison of count data between groups was conducted using the chi-square test or Fisher exact probability. The Kaplan-Meier method was used to calculate survival rate and plot survival curve, and Log-rank test was used for survival analysis. The Cox risk regression model was used for univariate and multivariate analyses. Results:(1) Clinicopatholo-gical characteristics of patients. None of the 36 patients with biliary NENs had carcinoid syndrome. There were 11 cases with tumor located at gallbladder, 14 cases with tumor located at bile duct, and 11 cases with tumor located at ampulla of Vater. There were significant differences in weight loss and TNM stage among biliary NENs patients with different tumor location ( χ2=9.14, 6.54, P<0.05). Of the 36 patients, there were 12 cases with neuroendocrine tumors, 16 cases with neuroendocrine carcinomas, and 8 cases with mixed neuroendocrine-non-neuroendocrine neoplasms. (2) Follow-up. All 36 patients were followed up for 39(range, 10-93)months. Of the 36 patients, 19 patients experienced tumor recurrence and 16 patients experienced tumor metastasis. There were 18 patients died. The median overall survival time of 36 patients was 30 months, with the 1-, 2-, 3-year overall survival rates of 63.9%, 51.0%, and 35.7%, respectively. The 1-, 2-, 3-year recurrence-free survival rates were 47.5%, 34.1% and 21.3%, respectively. The 1-, 2-, 3-year overall survival rates of 19 patients with tumor recurrence were 55.6%, 55.6% and 27.8%, respectively. The 1-, 2-, 3-year overall survival rates of 17 patients without tumor recurrence were 71.3%, 50.4% and 42.0%, respectively. There was no significant difference in overall survival between patients with and without tumor recurrence ( χ2=0.24, P>0.05). (3) Prognostic factors analysis of patients. Results of multivariate analysis showed that pathological type as neuroendocrine carcinomas and mixed neuroendocrine-non-neuroendocrine neoplasms, non-R 0 margin were independent risk factors influencing overall survival time of patients ( hazard ratio=5.50, 5.33, 14.04, 95% confidence interval as 1.32-23.01, 1.17-24.35, 2.67-73.79, P<0.05). Conclusions:Biliary NENs lack specific clinical manifestations. Poorly differentiated neuroendocrine carcinomas are the most common pathological type. Pathological type as neuroendocrine carcinomas and mixed neuroendocrine-non-neuroendocrine neoplasms, non-R 0 margin are independent risk factors influencing prognosis of patients.

Neuronal intranuclear inclusion disease（NIID）is a rare hereditary neurodegenerative disorder that can affect multiple systems. However，it is uncommon for urinary dysfunction to be the initial symptom. This article reports five cases. The five patients began to experience voiding dysfunction such as frequent urination，weak urination，and incomplete urination at the mean ages of 55.4（47 - 65）years old. Four months to twelve years after urinary onset，neurological symptoms such as headache，memory decline，transient loss of consciousness，and unsteady gait began to appear. Four of the five cases had a family history. Brain MRI revealed the “ribbon sign” or “crest sign” in all cases. Skin biopsy revealed eosinophilic inclusions in the cell nuclei，and NOTCH2NLC gene testing identified abnormal GGC mutations. Three of the five patients underwent cystostomy due to secondary hydronephrosis，while the other two received no special treatment. After a follow-up of 18 to 35 months since diagnosis，the patients who underwent cystostomy had normal renal function. Neurological symptoms in all five patients worsened to varying degrees.

Background and Aims:Tumor deposits(TDs)may influence prognosis beyond the current 8th edition AJCC pTNM nodal classification in gastric cancer(GC).This study investigates the prognostic value of TD number and proposes an improved pN staging(mpN)that classifies patients with TD number>1 as pN3b.We validated the mpN staging against the 8th AJCC pN staging.Methods:A dual-center retrospective cohort study was performed,including 1 327 patients who underwent radical gastrectomy at Sun Yat-sen University Cancer Center(2011-2015;test cohort)and 340 patients from Guangdong Provincial People's Hospital(2015-2022;validation cohort).Patients were dichotomized into low-TD(≤1)and high-TD(>1)groups.Outcomes were overall survival(OS)and disease-free survival(DFS).Survival analyses used Kaplan-Meier curves,IPTW,and Cox regression.Predictive performance of staging systems was assessed by time-dependent ROC(tROC)/tAUC,concordance index(C-index)and Akaike information criterion(AIC).Results:TDs were present in 435/1 327(32.7%)in the test cohort.Presence of TD was associated with worse OS(IPTW-adjusted HR=2.69,95%CI=2.18-3.31,P<0.01)and DFS(HR=2.82,95%CI=2.32-3.42,P<0.01).In multivariable models,TD remained an independent adverse factor for OS(HR=1.65,95%CI=1.34-2.05;P<0.01)and DFS(HR=1.74,95%CI=1.43-2.11,P<0.01).Increasing TD number correlated with progressively poorer survival;X-tile identified>1 as an optimal cutoff,with high-TD patients showing markedly worse outcomes(OS:adjusted HR=3.65,95%CI=2.74-4.88;DFS:adjusted HR=3.74,95%CI=2.85-4.91;both P<0.01).Incorporation of TD number into the mpN staging(assigning TD>1 to pN3b)improved prognostic discrimination:in the test cohort 5-year OS tAUC was 0.746 for mpN vs.0.703 for AJCC pN(C-index 0.738 vs.0.721,AIC 5 805.27 vs.5 849.30);similar improvements were observed in the validation cohort.Conclusion:TD presence and number exert significant negative prognostic impact in GC.Classifying patients with TD number>1 as pN3b enhances prognostic accuracy.Routine reporting of TD counts and further prospective multicenter validation of mpN staging are warranted.

Neuronal intranuclear inclusion disease（NIID）is a rare hereditary neurodegenerative disorder that can affect multiple systems. However，it is uncommon for urinary dysfunction to be the initial symptom. This article reports five cases. The five patients began to experience voiding dysfunction such as frequent urination，weak urination，and incomplete urination at the mean ages of 55.4（47 - 65）years old. Four months to twelve years after urinary onset，neurological symptoms such as headache，memory decline，transient loss of consciousness，and unsteady gait began to appear. Four of the five cases had a family history. Brain MRI revealed the “ribbon sign” or “crest sign” in all cases. Skin biopsy revealed eosinophilic inclusions in the cell nuclei，and NOTCH2NLC gene testing identified abnormal GGC mutations. Three of the five patients underwent cystostomy due to secondary hydronephrosis，while the other two received no special treatment. After a follow-up of 18 to 35 months since diagnosis，the patients who underwent cystostomy had normal renal function. Neurological symptoms in all five patients worsened to varying degrees.

Background and Aims:Tumor deposits(TDs)may influence prognosis beyond the current 8th edition AJCC pTNM nodal classification in gastric cancer(GC).This study investigates the prognostic value of TD number and proposes an improved pN staging(mpN)that classifies patients with TD number>1 as pN3b.We validated the mpN staging against the 8th AJCC pN staging.Methods:A dual-center retrospective cohort study was performed,including 1 327 patients who underwent radical gastrectomy at Sun Yat-sen University Cancer Center(2011-2015;test cohort)and 340 patients from Guangdong Provincial People's Hospital(2015-2022;validation cohort).Patients were dichotomized into low-TD(≤1)and high-TD(>1)groups.Outcomes were overall survival(OS)and disease-free survival(DFS).Survival analyses used Kaplan-Meier curves,IPTW,and Cox regression.Predictive performance of staging systems was assessed by time-dependent ROC(tROC)/tAUC,concordance index(C-index)and Akaike information criterion(AIC).Results:TDs were present in 435/1 327(32.7%)in the test cohort.Presence of TD was associated with worse OS(IPTW-adjusted HR=2.69,95%CI=2.18-3.31,P<0.01)and DFS(HR=2.82,95%CI=2.32-3.42,P<0.01).In multivariable models,TD remained an independent adverse factor for OS(HR=1.65,95%CI=1.34-2.05;P<0.01)and DFS(HR=1.74,95%CI=1.43-2.11,P<0.01).Increasing TD number correlated with progressively poorer survival;X-tile identified>1 as an optimal cutoff,with high-TD patients showing markedly worse outcomes(OS:adjusted HR=3.65,95%CI=2.74-4.88;DFS:adjusted HR=3.74,95%CI=2.85-4.91;both P<0.01).Incorporation of TD number into the mpN staging(assigning TD>1 to pN3b)improved prognostic discrimination:in the test cohort 5-year OS tAUC was 0.746 for mpN vs.0.703 for AJCC pN(C-index 0.738 vs.0.721,AIC 5 805.27 vs.5 849.30);similar improvements were observed in the validation cohort.Conclusion:TD presence and number exert significant negative prognostic impact in GC.Classifying patients with TD number>1 as pN3b enhances prognostic accuracy.Routine reporting of TD counts and further prospective multicenter validation of mpN staging are warranted.

Objective:To investigate the clinicopathological characteristics and prognostic factors of biliary neuroendocrine neoplasms (NENs).Methods:The retrospective cohort study was conducted. The clinicopathological data of 36 patients who underwent surgical treatment for biliary NENs at The First Hospital of Jilin University from January 2013 to December 2023 were collected. There were 22 males and 14 females, aged (59±9)years. Observation indicators: (1) clinicopatholo-gical characteristics of patients; (2) follow-up; (3) prognostic factors analysis of patients. Compari-son of measurement data with normal distribution among multiple groups was conducted using the ANOVA. Comparison of measurement data with skewed distribution among multiple groups was conducted using the Kruskal-Wallis H test. Comparison of count data between groups was conducted using the chi-square test or Fisher exact probability. The Kaplan-Meier method was used to calculate survival rate and plot survival curve, and Log-rank test was used for survival analysis. The Cox risk regression model was used for univariate and multivariate analyses. Results:(1) Clinicopatholo-gical characteristics of patients. None of the 36 patients with biliary NENs had carcinoid syndrome. There were 11 cases with tumor located at gallbladder, 14 cases with tumor located at bile duct, and 11 cases with tumor located at ampulla of Vater. There were significant differences in weight loss and TNM stage among biliary NENs patients with different tumor location ( χ2=9.14, 6.54, P<0.05). Of the 36 patients, there were 12 cases with neuroendocrine tumors, 16 cases with neuroendocrine carcinomas, and 8 cases with mixed neuroendocrine-non-neuroendocrine neoplasms. (2) Follow-up. All 36 patients were followed up for 39(range, 10-93)months. Of the 36 patients, 19 patients experienced tumor recurrence and 16 patients experienced tumor metastasis. There were 18 patients died. The median overall survival time of 36 patients was 30 months, with the 1-, 2-, 3-year overall survival rates of 63.9%, 51.0%, and 35.7%, respectively. The 1-, 2-, 3-year recurrence-free survival rates were 47.5%, 34.1% and 21.3%, respectively. The 1-, 2-, 3-year overall survival rates of 19 patients with tumor recurrence were 55.6%, 55.6% and 27.8%, respectively. The 1-, 2-, 3-year overall survival rates of 17 patients without tumor recurrence were 71.3%, 50.4% and 42.0%, respectively. There was no significant difference in overall survival between patients with and without tumor recurrence ( χ2=0.24, P>0.05). (3) Prognostic factors analysis of patients. Results of multivariate analysis showed that pathological type as neuroendocrine carcinomas and mixed neuroendocrine-non-neuroendocrine neoplasms, non-R 0 margin were independent risk factors influencing overall survival time of patients ( hazard ratio=5.50, 5.33, 14.04, 95% confidence interval as 1.32-23.01, 1.17-24.35, 2.67-73.79, P<0.05). Conclusions:Biliary NENs lack specific clinical manifestations. Poorly differentiated neuroendocrine carcinomas are the most common pathological type. Pathological type as neuroendocrine carcinomas and mixed neuroendocrine-non-neuroendocrine neoplasms, non-R 0 margin are independent risk factors influencing prognosis of patients.

Objective:To investigate the impact of the number of examined lymph nodes (ELN) on survival outcomes in gastric cancer patients with postoperative pathological stage pN3b.Methods:This retrospective cohort study included 279 pN3b gastric cancer patients who underwent D2 gastrectomy at Nanfang Hospital, Southern Medical University (September 2008 to April 2023), with 35 patients receiving combination chemotherapy and anti-PD-1 therapy (immunotherapy group) and 244 receiving adjuvant chemotherapy alone (nonimmunotherapy group). Additionally, 422 patients with pN3b from the SEER database (2005 to 2020) were collected as an external validation cohort to determine the optimal cutoff value for the number of lymph nodes examined in the nonimmunotherapy group. The primary endpoints were overall survival (OS) and recurrence-free survival (RFS) in the nonimmunotherapy group of the Nanfang Hospital cohort, stratified by whether the number of examined lymph nodes was above or below the ELN optimal cutoff value. These findings were subsequently validated in the SEER cohort.Results:The optimal ELN cutoff value (34 nodes) was determined using X-tile software and by constructing an ELN-HR fitting model with inflection point identification. In the nonimmunotherapy group, patients with ELN >34 exhibited significantly prolonged survival compared to ELN ≤34 (median OS: 25.0 (95%CI:20.5-29.5) to 17.0 (95%CI:12.7-21.3) months, P=0.004; median RFS: 19.0 (95%CI:15.6-22.4) to 13.0 (95%CI:9.5-16.5) months, P=0.048). Multivariate Cox analysis also showed ELN >34 to be an independent protective factor for both OS (HR=0.576, 95%CI: 0.397-0.836) and RFS (HR=0.701, 95%CI: 0.492-0.998). In the SEER cohort, ELN >34 was associated with a 5-month OS extension (19 to 14 months, P=0.065), with multivariate analysis supporting its independent prognostic significance (HR=0.729, 95%CI: 0.580-0.915, P=0.006). Notably, in the immunotherapy group, patients with ELN >34 ( n=30) achieved a median OS of 41 months, but the median OS had not been reached in the ELN ≤34 group ( n=5) (1 death at 48 months). Conclusion:Higher ELN (>34) correlates with improved survival in nonimmunotherapy-treated pN3b gastric cancer patients. However, in pN3b gastric cancer patients treated with immunotherapy, the optimal ELN threshold requires further exploration to determine.