Objective To construct a diagnostic and prognostic model for malignant pleural effusion (MPE) in patients with non-M1b stage (AJCC 7th edition) non-small cell lung cancer (NSCLC) by machine learning. Methods Retrospective analysis was conducted on patients diagnosed with NSCLC in the Surveillance, Epidemiology, and End Results database from 2010 to 2015, excluding those in the M1b stage. Two sets of data were collected: data 1 (patients with non-M1b stage NSCLC, n=47 392) was used to construct the MPE diagnostic model; and data 2 (patients with M1a stage NSCLC and MPE, n=2 422) was used to construct a prognostic model. The Least Absolute Shrinkage and Selection Operator (LASSO) regression was used to screen feature variables, with a training set and validation set ratio of 7:3. Models were built using eight machine learning algorithms, with evaluation metrics including accuracy, precision, recall, F1 score, area under the ROC curve (AUC), decision curve, calibration curve, and precision recall curve (PR), with ROC-AUC as the main evaluation metric. Results The incidence of MPE in patients with non-M1b stage NSCLC was 5.12%, and the 1-year survival rate of patients with MPE was 32.5%. LASSO regression identified nine diagnostic-related variables and 12 prognostic-related variables. The AUC values of the models constructed by eight machine learning algorithms all exceeded 0.70. The random forest model performed the best in the diagnostic model (training set AUC=0.908, validation set AUC=0.897), and the XGBoost model showed the best performance in the prognostic model (training set AUC=0.905, validation set AUC=0.875). Other evaluation indicators showed good results and balanced distribution. SHAP feature importance analysis showed that tumor size, lymph node metastasis, and histological type were important influencing factors for the occurrence of MPE, and chemotherapy intervention was the most remarkably prognostic factor. Conclusion The random forest diagnostic model constructed in this study can effectively predict the risk of MPE in patients with non-M1b stage NSCLC, and the XGBoost prognostic model can predict the prognosis of M1a-stage NSCLC patients with concurrent MPE.

Gut microbial communities are likely remodeled in tandem with accumulated physiological decline during aging, yet there is limited understanding of gut microbiome variation in advanced age. Here, we performed a metagenomics-based enterotype analysis in a geographically homogeneous cohort of 367 enrolled Chinese individuals between the ages of 60 and 94 years, with the goal of characterizing the gut microbiome of elderly individuals and identifying factors linked to enterotype variations. In addition to two adult-like enterotypes dominated by Bacteroides (ET-Bacteroides) and Prevotella (ET-Prevotella), we identified a novel enterotype dominated by Escherichia (ET-Escherichia), whose prevalence increased in advanced age. Our data demonstrated that age explained more of the variance in the gut microbiome than previously identified factors such as type 2 diabetes mellitus (T2DM) or diet. We characterized the distinct taxonomic and functional profiles of ET-Escherichia, and found the strongest cohesion and highest robustness of the microbial co-occurrence network in this enterotype, as well as the lowest species diversity. In addition, we carried out a series of correlation analyses and co-abundance network analyses, which showed that several factors were likely linked to the overabundance of Escherichia members, including advanced age, vegetable intake, and fruit intake. Overall, our data revealed an enterotype variation characterized by Escherichia enrichment in the elderly population. Considering the different age distribution of each enterotype, these findings provide new insights into the changes that occur in the gut microbiome with age and highlight the importance of microbiome-based stratification of elderly individuals.
Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Bacteroides ; China ; Diabetes Mellitus, Type 2/microbiology* ; Escherichia coli/classification* ; Gastrointestinal Microbiome/genetics* ; Metagenomics ; East Asian People

Portal vein thrombosis (PVT) is one of the most common complications of liver cirrhosis. The formation of PVT can increase the mortality rate of cirrhotic patients and adversely affect the successful implementation and prognosis of liver transplantation. A hypercoagulable state is a unique mechanism underlying PVT formation in cirrhotic patients. In recent years, the pathogenesis of PVT has gradually been elucidated, with specific mechanisms including the following aspects: systemic and local inflammatory responses lead to vascular endothelial cell dysfunction, thereby promoting the activation of the coagulation system; abnormal activation of the monocyte-macrophage system exacerbates local inflammation, enhancing platelet adhesion and aggregation, and facilitating thrombus formation; an imbalance between the coagulation and fibrinolytic systems results in a sustained hypercoagulable state; and intestinal microbiota dysbiosis induces inflammation and metabolic disturbances, thereby increasing the risk of PVT. This article summarizes the latest research progress on these key mechanisms and their interactions, providing new insights into the molecular and cellular mechanisms of PVT. It also offers directions for the early diagnosis of PVT and the exploration of novel intervention strategies in the future.

OBJECTIVE To develop a highly sensitive and specific quantitative real-time poly-merase chain reaction(qPCR)method for detecting the VGM-R02b(a gene therapy drug for glutaric acidemia type Ⅰ)gene in cynomolgus monkeys and analyze the biological distribution of VGM-R02b.METHODS A standard curve was constructed using the VGM-R02b standard plasmid[an adeno-associ-ated virus serotype 9(AAV9)capsid]on a qPCR platform.The detection method was optimized and validated for key parameters,including the quantitative range,accuracy,precision,dilution linearity,selectivity,specificity,stability,and parallelism.The established method was used to determine the target gene of VGM-R02b in the blood,brain,stomach,heart,liver,spleen,lung,kidney,thymus,and duodenum of cynomolgus monkeys on day 29(D29)and D92 after a single,unilateral intraventricular injection of VGM-R02b.The biodistribution of VGM-R02b in cynomolgus monkeys was analyzed.RESULTS A qPCR method for quantifying the VGM-R02b target gene in cynomolgus monkeys was established and validated.The standard curve demonstrated a quantitative range of 5.00×109 to 5.00×10^1 copies·μg-1 DNA,with excellent precision,accuracy,dilution linearity,selectivity,and specificity.The target gene in tissues remained stable after being stored at room temperature for 4 h,-15 to-25℃ for 9 d,-60 to-80℃ for 90 d and after five freeze-thaw cycles.Similarly,the target gene in whole blood remained stable after being stored at room temperature for 4 h,-15 to-25℃ for 93 d,-60 to-80℃ for 93 d and after five freeze-thaw cycles.Extracted nucleic acid samples also showed stability after being stored at room temperature for 4.25 h,2-8℃ for 24.16 h,-60 to-80℃ for 109 d and after five freeze-thaw cycles.The method was also applied to evaluate the biological distribution of the target gene in cynomolgus monkeys.In the control group,the target gene was undetectable on D29 and D92 post-administration,but in the drug administration group,the target gene was distributed across the tissues,with higher concen-trations observed in the brain,liver,spleen,and spinal cord,and there were no significant differences in the target gene content across the tissues between D29 and D92.CONCLUSION The established qPCR method is robust,reliable and suitable for determination of VGM-R02b target gene in cynomolgus monkeys.

Objective:To investigate the awareness levels of doctors from various departments regarding fertility preservation to provide reference and suggestions for further improving awareness and promoting multidisciplinary development in fertility preservation.Methods:From November to December 2023, the China Alliance of Fertility Preservation initiated this survey covering 88 hospitals in 23 cities across 20 provinces in China. The survey focused on the degree of understanding of basic concepts, technological perceptions, treatment concepts, and related challenges regarding fertility preservation. Descriptive statistics and Fisher's exact test were used to analyze the difference in awareness levels across various departments.Results:There was a significant difference in the awareness of fertility preservation guidelines among physicians from different departments ( P<0.001). Reproductive center physicians had the highest level of understanding [68.3% (41/60)]. Doctors in the reproductive centers had a higher awareness rate of the five fertility preservation techniques (ovarian tissue cryopreservation, oocyte cryopreservation, sperm cryopreservation, testicular tissue cryopreservation, and embryo cryopreservation) than that in other departments (all P<0.05). Regarding treatment concepts, more reproductive center physicians believed that psychological counselling about fertility preservation before oncological treatment was necessary ( P=0.016). Hematologists had more worries and concerns about fertility preservation treatment. Additionally, 96.7% (58/60) of reproductive center physicians expressed the need to establish effective referral mechanisms within the same city, province, or across provinces. However, 55.6% (35/63) of oncologists, 64.6% (31/48) of hematologists, and 77.8% (7/9) of pediatricians considered treating primary tumors far more important than fertility preservation. Conclusion:Doctors from different departments have varying levels of understanding regarding fertility preservation. It is necessary to strengthen the dissemination and training of fertility preservation knowledge among physicians from various departments, as well as to promote a multidisciplinary treatment model to improve efficiency, to establish a comprehensive referral mechanism to improve patients' opportunities for fertility preservation.

Objective:To investigate the impact of hysteroscopic surgery on fresh embryo transfer outcomes in patients with transvaginal ultrasound-diagnosed FIGO Type Ⅱ/Ⅲ uterine fibroids that may affect the uterine cavity morphology, providing a reference for clinical practice.Methods:A retrospective cohort study was performed to analyze the clinical data of 346 patients who visited the Reproductive Medicine Center, Department of Obstetrics and Gynecology, Peking University Third Hospital between January 2020 and December 2022, with transvaginal ultrasound findings indicating uterine fibroids adjacent to or protruding into the uterine cavity (FIGO Type Ⅱ, Ⅲ) and who underwent fresh embryo transfer. Patients were divided into two groups based on whether hysteroscopic surgery were performed: the hysteroscopic surgery group ( n=237, the group that underwent hysteroscopy before embryo transfer) and the non-surgery group ( n=109, the group that proceeded directly to embryo transfer without hysteroscopy). Basic patient characteristics and pregnancy outcomes were compared between the two groups. Multivariate logistic regression analysis was used to identify factors influencing clinical pregnancy rate and live birth rate. Results:The baseline characteristics showed no statistically significant differences between the two groups. The clinical pregnancy rate [44.3% (105/237)] and the live birth rate [32.5% (77/237)] following fresh embryo transfer in the hysteroscopic surgery group were significantly higher than those in the non-surgery group [31.2% (34/109), P=0.021; 18.3% (20/109), P=0.007], whereas the pregnancy loss rate showed no significant difference. Stratified analysis by age revealed that for patients aged <35 years, the clinical pregnancy rate [62.1% (54/87)] and the live birth rate [51.7% (45/87)] in fresh embryo transfer cycles were significantly higher in the hysteroscopic surgery group compared with the non-surgery group [30.8% (12/39), P=0.001; 25.6% (10/39), P=0.006]. Multivariate logistic regression analysis further confirmed that hysteroscopic surgery was an independent factor influencing live birth rate ( OR=2.128, 95% CI: 1.152-3.930, P=0.016). Among patients aged <35 years, hysteroscopic surgery was an influencing factor of both clinical pregnancy rate ( OR=4.222, 95% CI: 1.745-10.215, P=0.001) and live birth rate ( OR=3.449, 95% CI: 1.436-8.282, P=0.006). Conclusion:For infertile patients with ultrasound findings of uterine fibroids adjacent to or protruding into the uterine cavity, especially younger patients, hysteroscopy is recommended. It can improve pregnancy outcomes of fresh embryo transfer, increasing both the clinical pregnancy rate and the live birth rate.

Objective:To investigate the role of serotonergic (5-HT) neurons activation in the dorsal raphe nucleus (DRN) in heat-induced seizures and its influence on breathing patterns in Scn1a + /- mice. Methods:24-day-old male Scn1a + /- mice were used for experiments, and sudden unexpected death in epilepsy(SUDEP) model was established through heat induction for 5 consecutive days. (1) Fluoxetine intervention experiment: 20 male mice were randomly divided into model group( n=10) and fluoxetine group ( n=10) according the weight-matched method. The fluoxetine group received intraperitoneal injection of fluoxetine (10 mg/kg) 45 min before heat induction each day for 5 consecutive days, while the model group received equal volume of 0.9% NaCl solution. (2) Chemogenetic activation experiment: 20 male mice were randomly divided into vector control group( n=10) and chemogenetic activation group ( n=10) according the weight-matched method. Empty vector or rAAV-TPH2-hM3d(Gq)-EGFP-WPREs was stereotaxically injected into DRN 14 d prior to seizure induction, and deschloroclozapine (5 mg/kg) was intraperitoneally injected 30 min before heat induction. Seizure characteristics and survival were assessed through video monitoring, and respiratory parameters were monitored. Immunofluorescence was used to detect colocalization of tryptophan hydroxylase 2(TPH2) and c-Fos in DRN. Statistical analysis was performed using SPSS 24.0 and GraphPad Prism 9.0. The independent sample t-test or Mann-Whitney test was used for inter-group comparison. Results:(1) In the fluoxetine intervention experiment: the survival rates between the model group and fluoxetine group showed no statistically significant difference ( χ2=2.23, P>0.05). As for the frequency of grade Ⅳ seizures, the model group (1.50(1.25, 2.35)min) demonstrated higher frequency than the fluoxetine group (0.43(0.20, 0.67)min) ( t=-3.40, P<0.05).With respect to respiratory parameters, the model group demonstrated shorter expiratory time ((0.10±0.02) s) and inspiratory time ((0.15±0.02) s) compared to the fluoxetine group ((0.16±0.05) s, (0.19±0.04) s) ( t=-3.47, -3.73, both P<0.01). The respiratory rate in the model group ((269.96±44.84) times/min) was significantly higher compared to the fluoxetine group ((195.04±52.37) times/min) ( t=3.44, P<0.01). The tidal volume in the model group ((0.10±0.02) mL) was significantly lower than the fluoxetine group ((0.13±0.04) mL) ( t=-2.19, P<0.05).The number of TPH2+ /c-Fos+ co-expressing cells in the model group (11.00±4.00) was lower than that in the fluoxetine group (33.00±8.39)( t=-4.16, P<0.05). (2) In the chemogenetic activation experiment: compared to the vehicle group, the chemogenetic activation group demonstrated significantly enhanced survival rates ( χ2=5.83, P<0.05). As for the frequency of grade Ⅴ seizures, the vehicle group (2.11(1.62, 3.44) times/min) showed higher frequency compared to the chemogenetic activation group (0.81(0.00, 1.62) times/min) ( t=18.00, P<0.05). In terms of respiratory parameters, the vehicle group showed shorter expiratory time ((0.10±0.01) s) and inspiratory time ((0.14±0.01) s) compared to the chemogenetic activation group ((0.12±0.01) s, (0.15±0.01) s) ( t=-2.78, -2.50, both P<0.05). The respiratory rate in the vehicle group ((208.37±9.73) times/min) was significantly higher than the chemogenetic activation group ((191.85±8.83) times/min) ( t=3.98, P<0.01). The tidal volume in the vehicle group ((0.09±0.01) mL) was significantly lower than the chemogenetic activation group ((0.12±0.02) mL) ( t=-4.77, P<0.001).The number of TPH2+ /c-Fos+ co-expressing cells in the vehicle group (9.00±3.46) was lower than that in the chemogenetic activation group (43.00±11.02)( t=-5.20, P<0.01). Conclusion:Specific activation of serotonergic neurons in the DRN can ameliorate heat-induced epileptic symptoms, improve respiratory function, and prolong survival time in Scn1a + /- mice.

Objective To describe and evaluate the clinical studies of postoperative pain sensitization caused by sleep deprivation through the evidence map system,understand the distribution of evidence in this field,and provide reference for subsequent evidence research.Methods A computer-based search of PubMed,EMBASE,Cochrane library,Web of Science,CNKI,Wanfang Data,VIP and Chinese Biomedical Literature Database from inception to August 2023 was conducted to obtain intervent ion studies,observational studies and systematic reviews/Meta-analysis of postoperative pain sensitization caused by sleep deprivation.The research characteristics and methodological quality were analyzed and evaluated.The Cochrane Handbook for Systematic Reviews,the Newcastle-Ottawa Scale(NOS)and the AMSTAR-2 scale were used to evaluate the quality of the included studies,and the evidence was comprehensively analyzed and displayed by means of bubble chart,table and text.Results A total of 35 observational studies(31 cohort studies and 4 case-control studies),15 randomized controlled trials and 4 systematic reviews/Meta-analyses were included.The number of publications increased rapidly after 2018 and peaked in 2022,and clinical studies in this field mainly fo-cused on cohort studies,with fewer randomized controlled trials and systematic reviews/Meta-analysis studies.The results of the evidence map showed that in terms of quality,22 studies were'high quality',24 studies were'medium quality',and 8 studies were'low quality'.Thirty studies showed that sleep deprivation could induce postoperative pain sensitization.Only 2 studies suggested that sleep disorders were not significantly asso-ciated with postoperative pain sensitization,and ten studies were uncertain whether sleep deprivation could in-duce postoperative pain sensitization.Conclusions Overall evidence shows that sleep deprivation can induce postoperative pain sensitization,but the evaluation dimensions are limited and the methodological quality of the included literature needs to be improved.More high-quality,large-sample and standardized clinical studies should be carried out in the future to provide better scientific basis for clinical work.

Objective:To investigate the clinicopathological and molecular genetic characteristics of diffuse gliomas with the features of polymorphous low-grade neuroepithelial tumor of the young (PLNTY) and their prognostic values.Methods:A retrospective analysis was performed on 14 cases of diffuse gliomas with PLNTY features diagnosed at the First Affiliated Hospital of Fujian Medical University, Fuzhou, China from June 2020 to August 2024. Their clinicopathological characteristics were examined, and their molecular genetic and epigenetic features were assessed using next-generation sequencing (NGS) and methylation analysis. Factors influencing prognosis were also analyzed.Results:Among the 14 patients, there were 8 males and 6 females, aged 3-62 years, median 29 (9, 50) years. All cases were initially diagnosed as low-grade diffuse gliomas histologically but exhibited the histological and immunohistochemical features of PLNTY. At the molecular level, all cases showed molecular abnormalities involving the mitogen-activated protein kinase pathway, including 5 cases with FGFR3-TACC3 (F3T3) fusion, 3 cases with FGFR2 fusion, 5 cases with BRAF V600E mutation, and 1 case with FGFR1 mutation. Among them, TERT promoter mutations were frequently observed in tumors with F3T3 fusion (5/5), while NCOR2 in-frame insertion mutations were prominent in tumors with non-F3T3 fusions. Clinical follow-up showed recurrence in 3 cases, all of which had F3T3 fusion and concurrent TERT promoter mutations. Prognostic analysis confirmed that F3T3 fusion with concurrent TERT promoter mutation was associated with poor prognosis.Conclusions:Diffuse gliomas with PLNTY features exhibit heterogeneity in clinicopathology and molecular genetics, with FGFR3/FGFR2 fusions and BRAF/FGFR1 mutations as the most common molecular alteration. They often have concurrent F3T3 fusion and TERT promoter mutations, which are related to poor prognosis. The possibility of molecular glioblastoma should be considered for these tumors. It is thus recommended to perform genetic testing on diffuse gliomas with PLNTY features in order to facilitate integrated diagnosis and provide molecular evidence for accurate evaluation of prognoses.