BACKGROUND:Meningeal lymphatic vessels can drain cerebral spinal fluid and amyloid β-protein,promoting T lymphocyte to transport and home to deep cervical lymph nodes.A simple,quick and definite method of dural separation and accurate localization of deep cervical lymph nodes can provide strong support for the study of neurodegenerative diseases.OBJECTIVE:To establish a convenient and practical method for exfoliating dural and deep cervical lymph nodes.METHODS:ICR mice,3 months old,were taken,anesthetized and injected with Evans blue and tracer in the occipital pool for localizing deep cervical lymph nodes.A midline incision of about 3 cm in length was made about 5 mm above the clavicle,the superficial fat and fascia were bluntly separated,and the lateral sternocleidomastoid muscle was pulled to expose the deep cervical lymph nodes,which were removed under a stereomicroscope and frozen at-80℃.Subsequently,the mouse head was cut and the skin and muscles of the head were separated to expose the entire skull structure.The skull and brain tissue were separated from the foramen magnum along the lower parietal bone with scissors,and the complete skull top was obtained.The skull was sequentially fixed in 40 g/L paraformaldehyde solution for 24 hours,120 g/L paraformaldehyde for 24 hours,and 120 g/L paraformaldehyde for 10,20,30,and 40 minutes,and the dural structure was stripped.The drainage capacity of meningeal lymphatic vessels and deep cervical lymphatic vessels was verified by tracer,and the meningeal lymphatic vessels were identified by the lymphatic vessel endothelial hyaluronan receptor 1 using the immunofluorescence method.RESULTS AND CONCLUSION:(1)Obvious blue staining was observed in deep cervical lymph nodes 15 minutes after Evans blue staining.(2)The skull was sampled and fixed in 120 g/L paraformaldehyde for 24 hours,resulting in a less tight connection between the dura mater and the skull,and easier stripping of the dural structures with an intact shape.The dura mater fixed at 120 g/L concentration was more resilient and remained more intact during peeling compared with the conventional 40 g/L concentration;120 g/L paraformaldehyde fixed meninges for a short time,and 30-40 minutes was preferred.(3)The frozen section of deep cervical lymph nodes showed the presence of the tracer,complete meningeal lymphatic vessels were visible in the dura mater,and the tracer was observed at the tail of lymphatic vessels.Immunofluorescence staining for endothelial hyaluronan receptor 1 was positive in the deep cervical lymph nodes and dural lymphatics.In summary,the best peeling concentration and time is 120 g/L paraformaldehyde fixed for 24 hours.At this concentration,the dura mater has a stretched morphology,a better toughness,and is more intact after peeling,which is conducive to later use.Verified by Evans blue,tracers and immunofluorescence,deep cervical lymph nodes are located accurately,which can be used as a basis for the study of various neurodegenerative diseases.

Objective:To investigate the risk factors and their predictive efficacy for postoperative premature epiphyseal closure in pediatric patients with distal tibial epiphyseal fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 216 pediatric patients with distal tibial epiphyseal fractures admitted to Zhengzhou Orthopedics Hospital from January 2018 to December 2023, including 136 males and 80 females, aged 2-16 years [11.8(9.8, 13.3)years]. Among them, 112 patients were affected on the left side and 104 on the right. According to the Salter-Harris fracture classification, the fracture was classified as type I in 14 patients, type II in 97, type III in 38, type IV in 64 and type VI in 3. According to the presence of premature epiphyseal closure after surgery, the patients were divided into premature epiphyseal closure group ( n=38) and normal epiphyseal group ( n=178). Age, gender, body mass index (BMI), injury mechanism, side of injury, Salter-Harris classification of fracture, initial displacement distance of the fracture end, medial malleolar involvement by the fracture line surgical fixation method, operation duration, reduction method, and reduction quality were recorded in the two groups. Univariate analysis and binary Logistic regression analysis were used to evaluate and determine the independent risk factors for postoperative premature physeal closure in pediatric patients with distal tibial epiphyseal fracture. The receiver operating characteristic (ROC) curve was plotted and area under the curve (AUC) was calculated to evaluate the predictive efficacy of each risk factor for the occurrence of premature physeal closure in pediatric patients with distal tibial epiphyseal fractures. Results:Univariate analysis showed the occurrence of postoperative premature epiphyseal closure of the distal tibia was associated with age, Salter-Harris fracture classification, medial malleolar involvement by the fracture line, surgical fixation method, reduction method, and reduction quality ( P<0.05), while it was not correlated with gender, BMI, injury mechanism, side of injury, initial displacement distance of the fracture end, and operation duration ( P>0.05). Multivariate binary Logistic regression analysis showed that medial malleolar involvement by the fracture line ( OR=0.18, 95% CI 0.04, 0.76, P<0.05) and reduction quality ( OR=43.18,95% CI 10.71, 174.05, P<0.01) were significantly correlated with the occurrence of postoperative premature epiphyseal closure of the distal tibia. The results of ROC curve analysis showed that medial malleolar involvement by the fracture line had limited predictive efficiency (AUC=0.53, 95% CI 0.43, 0.63), reduction quality had moderate predictive efficacy (AUC=0.81, 95% CI 0.72, 0.91), while their combination demonstrated even higher predictive efficacy (AUC=0.83, 95% CI 0.74, 0.91). Conclusions:Medial malleolar involvement by the fracture line and reduction quality are independent risk factors for postoperative premature epiphyseal closure in pediatric patients with distal tibial epiphyseal fractures. Reduction quality demonstrates good predictive efficacy, while medial malleolar involvement by the fracture line shows limited predictive value. The combination of both factors achieves even better predictive performance.

Objective: To explore the relationship between beverage dependence and sleep quality among college students, providing empirical evidence for improving their sleep quality. Methods: From December 2024 to January 2025, a convenience sampling method was used to conduct a questionnaire survey among 3 974 college students from four universities in Anhui Province. The Beverage Addiction Scale for College Students (BASCS) was used to assess beverage dependence, and the Self rating Scale of Sleep(SRSS) was used to evaluate sleep quality. A multivariate Logistic regression model was employed to analyze the relationship between beverage dependence and sleep quality, and a restricted cubic spline model was used to examine the dose response relationship between the two. Results: The positive rate of beverage dependence symptoms among college students was 7.6%, with positive rates of 9.6%, 13.8%, and 7.4% for the withdrawal symptoms, health effects, and dependence symptoms dimensions, respectively. The detection rate of sleep disorders was 23.6%. Multivariate Logistic regression analysis showed that after adjusting for covariates such as grade, gender, and body mass index, compared with the no beverage dependence group, students with positive beverage dependence symptoms had a higher risk of sleep disorders( OR =3.71, 95% CI =2.87-4.80, P <0.01). The OR (95% CI ) for sleep disorders among students with positive symptoms in the withdrawal symptoms, health effects, and dependence symptoms dimensions were 2.80(2.22-3.53), 2.38(1.95-2.91), and 2.45(1.89-3.18)(all P <0.01). Further analysis using a restricted cubic spline model revealed that the overall beverage dependence score and its three dimensional scores were approximately linearly related to the risk of sleep disorders among college students (all nonlinear P >0.05). Conclusions Beverage dependence is associated with sleep quality among college students. Schools should take multiple approaches, such as health education on beverage awareness, to improve students sleep quality.

Objective:To investigate the clinical effectiveness of tumor lesion curettage and allogeneic bone grafting via the fovea of femoral head through the surgical hip dislocation approach in children and adolescents with femoral head chondroblastoma (FHCB).Methods:Case series study.The clinical data of 7 FHCB patients [3 males and 4 females with a mean age of 13.7 years (11-19 years)] treated at the Department of Pediatric Orthopedics, Zhengzhou Orthopedic Hospital from January 2016 to March 2023 were retrospectively analyzed.All patients underwent lesion curettage and allogeneic bone grafting via the fovea of femoral head through the surgical hip dislocation approach.The healing, local recurrence and complications were assessed via clinical and radiological tests.Functional outcomes were evaluated using the Musculoskeletal Tumour Society (MSTS) scoring system.A paired t-test was used to compare the MSTS scores before the operation and at the final follow-up. Results:All patients were followed up for 30 months on average (18-48 months).The average healing time for bone grafting was 4.9 (3-8) months.Local recurrence and serious postoperative complications such as avascular necrosis of femoral head, secondary osteoarthritis, ectopic ossification and nonunion of greater trochanter osteotomy end were not observed in all patients up to the last follow-up.The average MSTS score at the last follow-up was (29.14±0.45) points.Conclusions:Curettage and bone grafting via the fovea of femoral head through the surgical hip dislocation approach is effective and safe in the treatment of FHCB in children and adolescents.This method can maintain the integrity of the articular surface and growth plate, and also preserve the blood supply of the femoral head and the normal function of the hip joint, with satisfactory short-term effects.

BACKGROUND:Meningeal lymphatic vessels can drain cerebral spinal fluid and amyloid β-protein,promoting T lymphocyte to transport and home to deep cervical lymph nodes.A simple,quick and definite method of dural separation and accurate localization of deep cervical lymph nodes can provide strong support for the study of neurodegenerative diseases.OBJECTIVE:To establish a convenient and practical method for exfoliating dural and deep cervical lymph nodes.METHODS:ICR mice,3 months old,were taken,anesthetized and injected with Evans blue and tracer in the occipital pool for localizing deep cervical lymph nodes.A midline incision of about 3 cm in length was made about 5 mm above the clavicle,the superficial fat and fascia were bluntly separated,and the lateral sternocleidomastoid muscle was pulled to expose the deep cervical lymph nodes,which were removed under a stereomicroscope and frozen at-80℃.Subsequently,the mouse head was cut and the skin and muscles of the head were separated to expose the entire skull structure.The skull and brain tissue were separated from the foramen magnum along the lower parietal bone with scissors,and the complete skull top was obtained.The skull was sequentially fixed in 40 g/L paraformaldehyde solution for 24 hours,120 g/L paraformaldehyde for 24 hours,and 120 g/L paraformaldehyde for 10,20,30,and 40 minutes,and the dural structure was stripped.The drainage capacity of meningeal lymphatic vessels and deep cervical lymphatic vessels was verified by tracer,and the meningeal lymphatic vessels were identified by the lymphatic vessel endothelial hyaluronan receptor 1 using the immunofluorescence method.RESULTS AND CONCLUSION:(1)Obvious blue staining was observed in deep cervical lymph nodes 15 minutes after Evans blue staining.(2)The skull was sampled and fixed in 120 g/L paraformaldehyde for 24 hours,resulting in a less tight connection between the dura mater and the skull,and easier stripping of the dural structures with an intact shape.The dura mater fixed at 120 g/L concentration was more resilient and remained more intact during peeling compared with the conventional 40 g/L concentration;120 g/L paraformaldehyde fixed meninges for a short time,and 30-40 minutes was preferred.(3)The frozen section of deep cervical lymph nodes showed the presence of the tracer,complete meningeal lymphatic vessels were visible in the dura mater,and the tracer was observed at the tail of lymphatic vessels.Immunofluorescence staining for endothelial hyaluronan receptor 1 was positive in the deep cervical lymph nodes and dural lymphatics.In summary,the best peeling concentration and time is 120 g/L paraformaldehyde fixed for 24 hours.At this concentration,the dura mater has a stretched morphology,a better toughness,and is more intact after peeling,which is conducive to later use.Verified by Evans blue,tracers and immunofluorescence,deep cervical lymph nodes are located accurately,which can be used as a basis for the study of various neurodegenerative diseases.

Objective:To investigate the clinical effectiveness of tumor lesion curettage and allogeneic bone grafting via the fovea of femoral head through the surgical hip dislocation approach in children and adolescents with femoral head chondroblastoma (FHCB).Methods:Case series study.The clinical data of 7 FHCB patients [3 males and 4 females with a mean age of 13.7 years (11-19 years)] treated at the Department of Pediatric Orthopedics, Zhengzhou Orthopedic Hospital from January 2016 to March 2023 were retrospectively analyzed.All patients underwent lesion curettage and allogeneic bone grafting via the fovea of femoral head through the surgical hip dislocation approach.The healing, local recurrence and complications were assessed via clinical and radiological tests.Functional outcomes were evaluated using the Musculoskeletal Tumour Society (MSTS) scoring system.A paired t-test was used to compare the MSTS scores before the operation and at the final follow-up. Results:All patients were followed up for 30 months on average (18-48 months).The average healing time for bone grafting was 4.9 (3-8) months.Local recurrence and serious postoperative complications such as avascular necrosis of femoral head, secondary osteoarthritis, ectopic ossification and nonunion of greater trochanter osteotomy end were not observed in all patients up to the last follow-up.The average MSTS score at the last follow-up was (29.14±0.45) points.Conclusions:Curettage and bone grafting via the fovea of femoral head through the surgical hip dislocation approach is effective and safe in the treatment of FHCB in children and adolescents.This method can maintain the integrity of the articular surface and growth plate, and also preserve the blood supply of the femoral head and the normal function of the hip joint, with satisfactory short-term effects.

Objective:To investigate the risk factors and their predictive efficacy for postoperative premature epiphyseal closure in pediatric patients with distal tibial epiphyseal fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 216 pediatric patients with distal tibial epiphyseal fractures admitted to Zhengzhou Orthopedics Hospital from January 2018 to December 2023, including 136 males and 80 females, aged 2-16 years [11.8(9.8, 13.3)years]. Among them, 112 patients were affected on the left side and 104 on the right. According to the Salter-Harris fracture classification, the fracture was classified as type I in 14 patients, type II in 97, type III in 38, type IV in 64 and type VI in 3. According to the presence of premature epiphyseal closure after surgery, the patients were divided into premature epiphyseal closure group ( n=38) and normal epiphyseal group ( n=178). Age, gender, body mass index (BMI), injury mechanism, side of injury, Salter-Harris classification of fracture, initial displacement distance of the fracture end, medial malleolar involvement by the fracture line surgical fixation method, operation duration, reduction method, and reduction quality were recorded in the two groups. Univariate analysis and binary Logistic regression analysis were used to evaluate and determine the independent risk factors for postoperative premature physeal closure in pediatric patients with distal tibial epiphyseal fracture. The receiver operating characteristic (ROC) curve was plotted and area under the curve (AUC) was calculated to evaluate the predictive efficacy of each risk factor for the occurrence of premature physeal closure in pediatric patients with distal tibial epiphyseal fractures. Results:Univariate analysis showed the occurrence of postoperative premature epiphyseal closure of the distal tibia was associated with age, Salter-Harris fracture classification, medial malleolar involvement by the fracture line, surgical fixation method, reduction method, and reduction quality ( P<0.05), while it was not correlated with gender, BMI, injury mechanism, side of injury, initial displacement distance of the fracture end, and operation duration ( P>0.05). Multivariate binary Logistic regression analysis showed that medial malleolar involvement by the fracture line ( OR=0.18, 95% CI 0.04, 0.76, P<0.05) and reduction quality ( OR=43.18,95% CI 10.71, 174.05, P<0.01) were significantly correlated with the occurrence of postoperative premature epiphyseal closure of the distal tibia. The results of ROC curve analysis showed that medial malleolar involvement by the fracture line had limited predictive efficiency (AUC=0.53, 95% CI 0.43, 0.63), reduction quality had moderate predictive efficacy (AUC=0.81, 95% CI 0.72, 0.91), while their combination demonstrated even higher predictive efficacy (AUC=0.83, 95% CI 0.74, 0.91). Conclusions:Medial malleolar involvement by the fracture line and reduction quality are independent risk factors for postoperative premature epiphyseal closure in pediatric patients with distal tibial epiphyseal fractures. Reduction quality demonstrates good predictive efficacy, while medial malleolar involvement by the fracture line shows limited predictive value. The combination of both factors achieves even better predictive performance.

Objective:To investigate the changes in interaction proteome of NUS1 mutant R290C and their relations with pathogenicity of Lennox Gastaut syndrome (LGS). Methods:The wild-type and mutant NUS1(R290C) plasmids were constructed and transfected into human embryonic kidney HEK293T cells; 48 h after that, NUS1 protein expression in HEK293T cells was detected by Western blotting. Co-immunoprecipitation, silver nitrate staining, and proteomic analysis were used to analyze the proteins interacted with wild-type or mutant NUS1 and identify the differential interacting proteins. Enrichment analyses of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were performed to annotate the molecular function and signaling pathways involved in the differential proteins. DisGeNet database was used to analyze the association between differential proteins and human diseases. Protein-protein interaction (PPI) was used to analyze the interaction network of NUS1 with protein folding regulatory proteins (RTN4 and DHDDS) and developmental epileptic encephalopathy related proteins.Results:(1) There was no significant difference in NUS1 protein expression between the wild-type and mutant NUS1 transfected HEK293T cells ( t=0.536, P=0.620). (2) Compared with that with wild-type NUS1 plasmid, number of proteins interacting with mutant NUS1 plasmid was significantly reduced in the transfected cells; 310 differential interacting proteins were screened in the mutant NUS1. (3) GO and KEGG enrichment analyses showed that the differential proteins were mainly involved in protein folding reaction and translation regulation. (4) DisGeNet association analysis showed that the two most relevant proteins in the differential interacting proteins were associated with frontotemporal dementia and developmental epileptic encephalopathy. (5) PPI analysis showed that NUS1 may be involved in occurrence of neurological diseases such as LGS by affecting protein folding signaling pathways. Conclusion:NUS1 mutant R290C alters its interacting protein lineage and mediates the development of LGS and other neurological diseases probably by regulating protein folding-related signaling.

Underactive bladder (UAB) belongs to the category of "urinary retention" and "urinary incontinence" in Traditional Chinese Medicine (TCM). Its pathogenesis is based on spleen qi deficiency and kidney deficiency, while dampness heat and blood stasis are the symptoms, and the nature of the disease is basedon the deficiency in nature and excess in superficiality and simultaneous occurrence of deficiency and excess syndrome. TCM treatment of UAB is mainly based on spleen qi deficiency syndrome, kidney deficiency syndrome, water and heat syndrome, deficiency and blood stasis syndrome to be treated with oral taken of Chinese materia medica, or the use of acupuncture, moxibustion, acupoint application and other external treatments, or the combination of acupuncture and medicine, TCM combined with rehabilitation training and other comprehensive treatments. All methods can improve the urodynamic indexes of patients, relieve symptoms and improve the quality of life, with certain efficacy.

Objective:To analyze the law of TCM syndrome differentiation and treatment for type 2 diabetic kidney disease (T2DKD) stage Ⅳ based on literature research.Methods:Literature on type 2 diabetic kidney disease stage Ⅳ was retrieved from CNKI, WanFang data, VIP and SinoMed database. The retrieval time was from the establishment of the databases to December 31, 2020. Data screening was conducted based on the inclusion and exclusion criteria prior to data entry in Microsoft Office Excel 365. Data mining and statistical analysis were performed by SPSS Statistics 23.0 and SPSS Modeler 18.1.Results:A total of 110 articles with 3 969 T2DKD stage Ⅳ cases, 111 prescriptions and 206 kinds of Chinese materia medica were included. Kidney and spleen were the main location of T2DKD stage Ⅳ. T2DKD stage Ⅳ based on TCM deficiency in nature syndrome was mainly based on qi and yin deficiency, and the most common excess in superficiality syndrome was blood stasis. The prescriptions commonly used included Liuwei Dihuang Decoction, Zhenwu Decoction, Buyang Huanwu Decoction, and Shenqi Dihuang Decoction etc. The classification of medication efficacy with the highest frequency was qi-tonifying herb, followed by blood-activating and stasis-resolving herb. Among them, Astragali Radix was the core Chinese materia medica in the prescription. The results of association rule obtained 54 association rules. Conclusions:The disease characteristics of T2DKD stage Ⅳ is simultaneous occurrence of deficiency and excess syndromes. The deficiency in nature is mainly characterized by deficiency of qi and yin, deficiency of spleen and kidney, deficiency of spleen-kidney yang, and excess in superficiality is mainly characterized by blood stasis, dampness and toxin. Tonifying qi and nourishing yin, activating blood circulation and dredging collaterals are the basic treatment methods, while strengthening spleen and kidney, dampness and detoxification should be emphasized. Astragali Radix, Angelicae Sinensis Radix, Salviae Miltiorrhizae Radix et Rhizoma, Poria, Dioscoreae Rhizoma, Corni Fructus, Rhei Radix et Rhizoma and Alismatis Rhizoma were the basic Chinese materia medica in this period, which reflects the idea of "treating qi, blood and water together".