Objective:To establish a rapid,quantitative and visualized method for the rapid detection of Streptococcus mutans(S.mutans)based on loop-mediated isothermal amplification(LAMP).Methods:Nucleic acid of S.mutans was extracted for LAMP reaction,the detection experimental conditions were optimized.Finally the positive results of the experiments were quantita-tively analyzed by using the mobile-based detection equipment.Results:The system detected the samples within 23 min and showed good specificity and sensitivity with an accuracy of 1.625×10-3 ng/μL.Positive results were captured,identified and quantified u-sing a mobile application,and the quantified results showed a good linear relationship between the absolute G value and the DNA concentration,with an equation of y=3.2x+57.133(R2=0.988 2).Conclusion:Quantitative detection of S.mutans by LAMP has the characteristics of high sensitivity,high specificity,result visualization,convenience and time-saving,and can quantify bacterial DNA.It is a new method for the early detection of caries susceptibility,and can be widely used in clinic.

Objective:This study analyzes the clinicopathological features and prognostic factors of early-stage gastric mixed adenoneuroendocrine carcinoma (G-MANEC), which is an exceedingly rare malignancy, in an effort to provide evidence-based insights for clinical decision-making.Methods:A retrospective observational study was conducted using the clinical data of 36 patients with early-stage G-MANEC who underwent surgical resection at Zhongshan Hospital, Fudan University, from July 2014 to May 2022. The observed indicators included clinicopathological data and follow-up information on recurrence, metastasis, and overall survival (OS).Results:Among the 36 patients there were 21 males and 15 females, aged 32-84 (65±11) years. The most common initial symptoms were abdominal pain and distension (19/36, 52.8%), followed by incidental findings during physical examinations (7/36, 19.4%). Tumors were located in the proximal stomach in 13 cases (36.1%), the middle stomach in 4 cases (11.1%), and the distal stomach in 19 cases (52.8%). Average tumor diameter was (2.48±1.18) cm. Gross morphology included elevated type in 12 cases (33.3%), flat type in 20 cases (55.6%), and depressed type in 4 cases (11.1%). Ulceration was present in 12 cases (33.3%). There were 11 cases (30.6%) at T1a stage and 25 cases (69.4%) at T1b stage. Lymph node metastasis was positive in 10 cases (27.8%), and the differentiation grades of the adenocarcinoma component were Grade I, II, and III in 3 (8.3%), 10 (27.8%), and 23 (63.9%) cases, respectively. Furthermore, the proportion of neuroendocrine carcinoma component was ≥50% in 18 cases (50.0%) and <50% in 18 cases (50.0%). Lymphovascular or perineural invasion was present in 18 cases (50.0%). Lauren classification included mixed type in 10 cases (27.8%), intestinal type in 19 cases (52.8%), and diffuse type in 7 cases (19.4%), and chromogranin A (CgA) positivity was found in 20 cases (55.6%). Additionally, the Ki-67 index positivity was found in 26 cases (72.2%). Total gastrectomy was performed in 12 cases (33.3%) and partial gastrectomy in 24 cases (66.7%), with a median follow-up duration of 77.5 months. The 3-year and 5-year OS rates were 88.89% and 79.67%, respectively. Univariate analysis revealed that age, gross morphology, ulceration, proportion of neuroendocrine carcinoma component, lymphovascular or perineural invasion, and chromogranin A (CgA) positivity showed statistical significance in their association with OS ( P<0.10). Multivariate Cox regression analysis further identified ulceration (HR=7.74, 95%CI: 1.24-48.30, P=0.028) and CgA positivity (HR=21.76, 95%CI: 1.86-53.97, P=0.014) as independent risk factors of OS. Conclusions:Patients with early-stage G-MANEC are typically asymptomatic, and those with ulceration or positive CgA immunohistochemical staining tend to have a poor prognosis.

Objective·To investigate the changes in transcriptome and chromatin accessibility during the differentiation of human embryonic stem cells(hESCs)into neural progenitor cells(NPCs)using in vitro differentiation models and high-throughput multi-omics sequencing technologies.Methods·hESCs were first induced to differentiate into NPCs in vitro using the embryoid body formation method,and cells at both stages were collected.The cell phenotypes were identified by reverse transcription-quantitative real-time PCR(RT-qPCR)and immunofluorescence(IF)staining.Transcriptome sequencing(RNA-seq)was conducted to detect and analyze the differentially expressed genes(DEGs)between hESCs and NPCs.The assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)was employed to assess chromatin accessibility changes between hESCs and NPCs.Motif enrichment analysis was performed on differentially accessible chromatin regions to discover potential regulatory transcription factors.Finally,an integrated analysis of RNA-seq and ATAC-seq data and the protein-protein interaction(PPI)network were performed to identify key genes and regulatory pathways involved in the early stages of neural differentiation in vitro.Results·Both RT-qPCR and IF results indicated that the expression levels of pluripotency markers(NANOG and POU5F1)were high at the hESC stage but significantly decreased at the NPC stage,while early neural differentiation markers(PAX6,SOX1,and NES)were minimally expressed at the hESC stage but markedly upregulated at the NPC stage.RNA-seq analysis revealed that compared to the hESC stage,there were 5 597 genes upregulated and 3 654 genes downregulated at the NPC stage.Gene function enrichment analysis showed that the upregulated genes at the NPC stage were enriched in the functions related to neural development.ATAC-seq analysis demonstrated a total of 27 491 genomic regions had significant changes in chromatin accessibility during the differentiation from hESC to NPC,with 12 381 regions showing increased accessibility and 15 110 regions showing decreased accessibility.Motif enrichment analysis revealed that transcription factor genes such as DLX1 and LHX2 might play an important role in the differentiation process from hESCs into NPCs.Integrated analysis of RNA-seq and ATAC-seq data revealed that overlapping genes with high expression at the NPC stage were mainly enriched in axon guidance,forebrain development,and neuron migration.After neural differentiation,the expression levels of CTNND2 and LHX2 genes increased,and the chromatin accessibility of related genomic regions also increased.PPI network analysis indentified candidate downstream genes including PRKACA,CDH2,and ERBB4.Conclusion·The in vitro differentiation model of hESCs combined with high-throughput multi-omics sequencing technologies can be used to depict the changes in transcriptome and chromatin accessibility during the differentiation of hESCs into NPCs.In this process,the expression levels of genes related to axon guidance,forebrain development,and neuronal migration pathways increase and related chromatin accessibility is enhanced.

This study evaluates the safety and early clinical outcomes of a novel 3D-printed titanium alloy "sleeve" component for revising fractured femoral stem prostheses in distal femoral megaprostheses without removing the fractured stem. The six patients included 2 males and 4 females, with an age range of 8-57 years. They were treated at Peking University People's Hospital between August 2020 and December 2023 and underwent revision surgery using the customized sleeve. A self-designed 3D-printed titanium alloy "sleeve" component was used for revision without removing the fractured stem, in the form of an external sleeve around the stem. Postoperative imaging was performed every three months to assess implant stability and bone integration. Functional outcomes were evaluated using the Musculoskeletal Tumor Society (MSTS)-93 score. All six patients successfully completed the surgery and follow-up, with surgical durations ranging from 120 to 230 minutes and intraoperative blood loss ranging from 150 to 800 ml. The follow-up period ranged from 6 to 46 months. At three months postoperatively, X-ray and CT imaging showed cortical bridging between the host bone and the "sleeve" component. By six months, full integration of the host cortical bone with the metal trabecular interface of the "sleeve" was observed. At the final follow-up, MSTS-93 scores ranged from 26 to 29 points, with no complications such as wound healing issues, implant loosening, fracture, infection, or degenerative arthritis. These findings suggest that 3D-printed titanium "sleeve" provide an effective, bone-preserving solution for femoral stem revision in oncologic megaprostheses, leading to favorable early stability and functional recovery.

Objective:To establish a rapid,quantitative and visualized method for the rapid detection of Streptococcus mutans(S.mutans)based on loop-mediated isothermal amplification(LAMP).Methods:Nucleic acid of S.mutans was extracted for LAMP reaction,the detection experimental conditions were optimized.Finally the positive results of the experiments were quantita-tively analyzed by using the mobile-based detection equipment.Results:The system detected the samples within 23 min and showed good specificity and sensitivity with an accuracy of 1.625×10-3 ng/μL.Positive results were captured,identified and quantified u-sing a mobile application,and the quantified results showed a good linear relationship between the absolute G value and the DNA concentration,with an equation of y=3.2x+57.133(R2=0.988 2).Conclusion:Quantitative detection of S.mutans by LAMP has the characteristics of high sensitivity,high specificity,result visualization,convenience and time-saving,and can quantify bacterial DNA.It is a new method for the early detection of caries susceptibility,and can be widely used in clinic.

Objective To analyze the clinicopathological characteristics and survival prognosis of young female patients with gastric adenocarcinoma. Methods A retrospective analysis was conducted on female patients who underwent radical gastrectomy at Zhongshan Hospital, Fudan University between January 2014 and December 2020, with postoperative pathological confirmation of gastric adenocarcinoma. Those aged ≤45 years were defined as the young group (n＝287), and were matched in a 1∶2 ratio by pTNM stage with female patients aged ≥60 years (elderly group, n＝574). Clinicopathological characteristics were compared between the two groups. Survival curves were plotted using the Kaplan-Meier method, and overall survival (OS) rates were assessed by log-rank test. Prognostic factors in the young group were analyzed using Cox regression models. Results Compared to elderly patients, young female gastric cancer patients exhibited a higher prevalence of tumors in the middle third of the stomach and a lower proportion in the upper third of the stomach. Molecular profiling revealed a higher frequency of HER2-low expression and elevated Ki-67 index. Pathologically, these patients were more frequently diagnosed with poorly differentiated or undifferentiated adenocarcinoma and signet ring cell carcinoma, while Lauren classification showed a predominance of the diffuse type with a lower proportion of the intestinal type (P＜0.05). Stratified analysis showed no significant difference in OS rates between the two groups for stage Ⅱ and Ⅲ patients; however, among stage Ⅰ patients, the young group had significantly better OS rates than the elderly group (P＝0.037). Multivariate Cox analysis and log-rank test confirmed that pN₃ stage (HR＝3.576, 95%CI 1.652–7.740), stage Ⅲ (HR＝3.581, 95%CI 1.059–12.106), and diffuse type (HR＝2.711, 95%CI 1.316–5.585) were risk factors for poor prognosis in young female gastric cancer patients. Conclusions Young female patients with gastric adenocarcinoma typically present with clinicopathological features such as the diffuse type, poor differentiation, and high proliferation. Moreover, pN₃ stage, stage Ⅲ cancer, and the diffuse type histology are correlated with a poor prognosis in this demographic.

This study evaluates the safety and early clinical outcomes of a novel 3D-printed titanium alloy "sleeve" component for revising fractured femoral stem prostheses in distal femoral megaprostheses without removing the fractured stem. The six patients included 2 males and 4 females, with an age range of 8-57 years. They were treated at Peking University People's Hospital between August 2020 and December 2023 and underwent revision surgery using the customized sleeve. A self-designed 3D-printed titanium alloy "sleeve" component was used for revision without removing the fractured stem, in the form of an external sleeve around the stem. Postoperative imaging was performed every three months to assess implant stability and bone integration. Functional outcomes were evaluated using the Musculoskeletal Tumor Society (MSTS)-93 score. All six patients successfully completed the surgery and follow-up, with surgical durations ranging from 120 to 230 minutes and intraoperative blood loss ranging from 150 to 800 ml. The follow-up period ranged from 6 to 46 months. At three months postoperatively, X-ray and CT imaging showed cortical bridging between the host bone and the "sleeve" component. By six months, full integration of the host cortical bone with the metal trabecular interface of the "sleeve" was observed. At the final follow-up, MSTS-93 scores ranged from 26 to 29 points, with no complications such as wound healing issues, implant loosening, fracture, infection, or degenerative arthritis. These findings suggest that 3D-printed titanium "sleeve" provide an effective, bone-preserving solution for femoral stem revision in oncologic megaprostheses, leading to favorable early stability and functional recovery.

Objective·To investigate the changes in transcriptome and chromatin accessibility during the differentiation of human embryonic stem cells(hESCs)into neural progenitor cells(NPCs)using in vitro differentiation models and high-throughput multi-omics sequencing technologies.Methods·hESCs were first induced to differentiate into NPCs in vitro using the embryoid body formation method,and cells at both stages were collected.The cell phenotypes were identified by reverse transcription-quantitative real-time PCR(RT-qPCR)and immunofluorescence(IF)staining.Transcriptome sequencing(RNA-seq)was conducted to detect and analyze the differentially expressed genes(DEGs)between hESCs and NPCs.The assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)was employed to assess chromatin accessibility changes between hESCs and NPCs.Motif enrichment analysis was performed on differentially accessible chromatin regions to discover potential regulatory transcription factors.Finally,an integrated analysis of RNA-seq and ATAC-seq data and the protein-protein interaction(PPI)network were performed to identify key genes and regulatory pathways involved in the early stages of neural differentiation in vitro.Results·Both RT-qPCR and IF results indicated that the expression levels of pluripotency markers(NANOG and POU5F1)were high at the hESC stage but significantly decreased at the NPC stage,while early neural differentiation markers(PAX6,SOX1,and NES)were minimally expressed at the hESC stage but markedly upregulated at the NPC stage.RNA-seq analysis revealed that compared to the hESC stage,there were 5 597 genes upregulated and 3 654 genes downregulated at the NPC stage.Gene function enrichment analysis showed that the upregulated genes at the NPC stage were enriched in the functions related to neural development.ATAC-seq analysis demonstrated a total of 27 491 genomic regions had significant changes in chromatin accessibility during the differentiation from hESC to NPC,with 12 381 regions showing increased accessibility and 15 110 regions showing decreased accessibility.Motif enrichment analysis revealed that transcription factor genes such as DLX1 and LHX2 might play an important role in the differentiation process from hESCs into NPCs.Integrated analysis of RNA-seq and ATAC-seq data revealed that overlapping genes with high expression at the NPC stage were mainly enriched in axon guidance,forebrain development,and neuron migration.After neural differentiation,the expression levels of CTNND2 and LHX2 genes increased,and the chromatin accessibility of related genomic regions also increased.PPI network analysis indentified candidate downstream genes including PRKACA,CDH2,and ERBB4.Conclusion·The in vitro differentiation model of hESCs combined with high-throughput multi-omics sequencing technologies can be used to depict the changes in transcriptome and chromatin accessibility during the differentiation of hESCs into NPCs.In this process,the expression levels of genes related to axon guidance,forebrain development,and neuronal migration pathways increase and related chromatin accessibility is enhanced.

Objective:This study analyzes the clinicopathological features and prognostic factors of early-stage gastric mixed adenoneuroendocrine carcinoma (G-MANEC), which is an exceedingly rare malignancy, in an effort to provide evidence-based insights for clinical decision-making.Methods:A retrospective observational study was conducted using the clinical data of 36 patients with early-stage G-MANEC who underwent surgical resection at Zhongshan Hospital, Fudan University, from July 2014 to May 2022. The observed indicators included clinicopathological data and follow-up information on recurrence, metastasis, and overall survival (OS).Results:Among the 36 patients there were 21 males and 15 females, aged 32-84 (65±11) years. The most common initial symptoms were abdominal pain and distension (19/36, 52.8%), followed by incidental findings during physical examinations (7/36, 19.4%). Tumors were located in the proximal stomach in 13 cases (36.1%), the middle stomach in 4 cases (11.1%), and the distal stomach in 19 cases (52.8%). Average tumor diameter was (2.48±1.18) cm. Gross morphology included elevated type in 12 cases (33.3%), flat type in 20 cases (55.6%), and depressed type in 4 cases (11.1%). Ulceration was present in 12 cases (33.3%). There were 11 cases (30.6%) at T1a stage and 25 cases (69.4%) at T1b stage. Lymph node metastasis was positive in 10 cases (27.8%), and the differentiation grades of the adenocarcinoma component were Grade I, II, and III in 3 (8.3%), 10 (27.8%), and 23 (63.9%) cases, respectively. Furthermore, the proportion of neuroendocrine carcinoma component was ≥50% in 18 cases (50.0%) and <50% in 18 cases (50.0%). Lymphovascular or perineural invasion was present in 18 cases (50.0%). Lauren classification included mixed type in 10 cases (27.8%), intestinal type in 19 cases (52.8%), and diffuse type in 7 cases (19.4%), and chromogranin A (CgA) positivity was found in 20 cases (55.6%). Additionally, the Ki-67 index positivity was found in 26 cases (72.2%). Total gastrectomy was performed in 12 cases (33.3%) and partial gastrectomy in 24 cases (66.7%), with a median follow-up duration of 77.5 months. The 3-year and 5-year OS rates were 88.89% and 79.67%, respectively. Univariate analysis revealed that age, gross morphology, ulceration, proportion of neuroendocrine carcinoma component, lymphovascular or perineural invasion, and chromogranin A (CgA) positivity showed statistical significance in their association with OS ( P<0.10). Multivariate Cox regression analysis further identified ulceration (HR=7.74, 95%CI: 1.24-48.30, P=0.028) and CgA positivity (HR=21.76, 95%CI: 1.86-53.97, P=0.014) as independent risk factors of OS. Conclusions:Patients with early-stage G-MANEC are typically asymptomatic, and those with ulceration or positive CgA immunohistochemical staining tend to have a poor prognosis.

Objectives:To investigate the survival and tumor recurrence after en bloc spondylectomy of spinal tumor and analyze the risk factors of postoperative tumor recurrence.Methods:This is a retrospective case series study. Data of 101 patients undergoing en bloc spondylectomy of spinal tumors in the Musculoskeletal Tumor Center, Peking University People′s Hospital from December 2006 to June 2022 were analyzed. There were 58 males and 43 females, aged (38.2±15.8) years (range: 10 to 79 years) at the time of surgery; the follow-up time was(44.0±36.0) months (range: 12 to 171 months).Among them, there were 25 relapsed patients, with 7 females and 18 males; aged (34.8±16.3) years (range: 12 to 66 years) at the time of surgery. The types of tumors included 5 giant cell tumors of bone, 6 osteosarcomas, 1 chordoma, 5 chondrosarcomas, 1 undifferentiated sarcoma, 1 fibrosarcoma, 2 Ewing sarcomas, 3 metastases and 1 malignant giant cell tumor of bone. Survival analysis of overall and relapsed patients were performed using the Kaplan-Meier curves. A segmented regression model was used to fit the sequence of recurrence rate changes over time since admission and identify change points for further analysis on risk factors. Univariate and multivariate Logistic regression analysis were performed to assess risk factors associated with recurrence rate; results from multivariate regression analysis were presented using a forest plot.Results:The tumor recurrence rate after en bloc spondylectomy was 24.8% (25/101).The overall median recurrence-free survival after en bloc spondylectomy was 161 months (95% CI: 92 months to NA).The median recurrence-free survival of recurrent patients was 13 months (95% CI: 12 to 27 months).Regarding the classification based on tumor malignancy, and relapse-free survival of metastatic tumors was significantly shorter ( P=0.007); and among the surgical margin groups, relapse-free survival of R0 group was significantly better than the R1 and R2 groups ( P<0.01). According to the segmented regression model, the tumor recurrence rate for en bloc spondylectomy showed a significant downward trend over time, with relatively higher recurrence rates before 2009 and a relatively stable trend after 2014. The results of univariate analysis showed that surgical margin and time of admission were the influencing factors of patient recurrence. The results of multivariate analysis showed that the R1 resection( OR=13.453,95% CI:2.897 to 97.941, P=0.002) and R2 resection( OR=11.379,95% CI:2.658 to 79.429, P=0.003) were independent influencing factor affecting patient recurrence. Conclusions:The overall tumor recurrence rate after en bloc spondylectomy was high. The surgical margin of tumor resection is an independent risk factor affecting tumor recurrence. Specifically, R2 and R1 resections significantly increase the risk of tumor recurrence.