Objective To construt the adenovirus vectors of Ad-hTERT-PUMA-EGFP containing human telomerase reverse transcriptase(hTERT) ,PUMA and enhanced green fluorescent protein (EGFP) .Methods By a series of molecular biology tech-niques ,PUMA gene was inserted into downstream of the hTERT promoter .And then EGFP was inserted into downstream of the PUMA in adenovirus vectors .Finally ,recombinant adenovirus vector ,Ad-hTERT-PUMA-EGFP was assembled into the adenovi-rus .Results Correct adenovirus laoding vectors was verified by viral genome PCR technology .After 12 h of transfection ,HEK293 cells showed green fluorescence in 72 h .Conclusion The correct structure of the adenovirus vectors containing PUMA gene and EGFP gene was identified by restriction enzyme analysis .

Objective To screen and identify differentially expressed genes in two new human urothelial carcinoma cell lines, BLS-211 and BLX. Methods Suppression subtractive hybridization (SSH) was used to createa subtracted library, and clones were sequenced. Results Totally 13 over-expressed genes in BLX and 9 in BLS-211 cells were obtained, respectively. Among them, 18 were known genes and 4 were new ESTs (Expressed Sequence Tag), and were collected by GenBank dbEST database (The access number was EB390424-7). Conclusion SSH is a powerful method for the identification of differentially expressed genes. The differential expression of some BCG-associated genes in different cells may be related to the different responses to clinical BCG therapy. The identified new ESTs can be cloned for full length to further study their functions.

Objective:To prepare the egg yolk immunoglobulin Y ( IgY) against human Sucrase and study its stability,in vitro activity. Methods:Hy-line laying hens were immunized with human Sucrase protein,IgY was isolated and purified from egg yolks of im-munized hens using water dilution and salting out method. Indirect ELISA was used to evaluate the titer and stability of IgY. The purity and specificity of IgY were analysed by SDS-PAGE and Western blot respectively. The inhibitory effects of IgY on α-glucosidase was studied by PNPG method. Results:Indirect ELISA results showed IgY could be detected on the tenth day after the first immunization, and the peak titer of IgY was 1:12 800 after the 40th day of immunization. SDS-PAGE showed that the heavy chain and light chain of IgY were 65 kD and 25 kD respectively, and the IgY against human Sucrase could specifically recognize the protein of human Sucrase. The IgY maintained primary titer when it was kept between 29-69℃ for 15 min,and pH 4-7,37℃,4 h. The titer of IgY was maintained 50% after digestion by pepsin and trypsin respectively for 2 hours. IgY had a higher resistence to pepsin than trypsin after longer digestion time. IgY showed an inhibitory effect on α-glucosidase in concentration dependent manner. The half inhibitory concentration (IC50) was 0. 540 mg/ml. Conclusion:The IgY against human Sucrase has been successfully obtained,which established foundations for its study of Type 2 diabetes mellitus rat models in vivo.

Spinal cord injuries is an extremely serious central nervous system injury. The clinical prognosis is very poor. Patients are often associated with lifelong disabilities or paralysis. Regulating NSCs to repair spinal cord injuries is unanimously considered to be a very potential option for the treatment of this type of disease. China has a large population and a large number of patients with spinal cord injuries. Actively regulating spinal cord NSCs is of great significance for the regeneration and repair of spinal cord injuries. Endogenous NSCs avoid many disadvantages of exogenous stem cell transplantation, and have a broader prospect in the treatment of spinal cord injuries. The Wnt signaling pathway plays a very important role in the differentiation of NSCs and the development of the nervous system. However, the molecular mechanism of the proliferation and differentiation of NSCs during regeneration and repair after spinal cord injuries is still not fully understood. This article mainly describes the research progress of Wnt pathway regulating NSCs in the regeneration and repair of spinal cord injuries.

Three-dimensional tooth segmentation is the segmentation of single-tooth models from a digital dental model. It is an important foundation for diagnosis, planning, treatment and customized appliance manufacturing in digital orthodontics. With the deep integration of artificial intelligence technology and big data from stomatology, the use of deep learning algorithms to assist 3D tooth segmentation has gradually become mainstream. This review summarizes the current situation of deep learning algorithms that assist 3D tooth segmentation from the aspects of dataset establishment, algorithm architecture, algorithm performance, innovation and advantages, deficiencies of current research and prospects. The results of the literature review showed that deep learning tooth segmentation methods could obtain an accuracy of more than 95% and had good robustness. However, the segmentation of complex dental models, operation time and richness of the training database still need to be improved. Research and development of the "consumption reduction and strong core" algorithm, establishment of an authoritative data sample base with multiple centers, and expansion of data application depth and breadth will lead to further development in this field.

ObjectiveTo investigate the current landscape and hotspots on researches about treatment of prolonged disorder of consciousness (pDOC) in the recent five years, and forecast the trends. MethodsLiterature about treatment of pDOC was retrieved from the Web of Science Core Collection database, from January 1st, 2019, to June 7th, 2023. The data were analyzed with CiteSpace 5.8.R3 to create knowledge maps for authors, countries, institutions, keywords, references, co-cited authors and co-cited literature. ResultsA total of 411 articles were included. Aurore Thibaut was the most influential author, Belgium was the most influential country, and Harvard Medical School was the institution with the most publications. The researches focused on neuromodulation, prognostic assessment and care, and management of swallowing function. The neuromodulation techniques mainly included transcranial direct current stimulation, repetitive transcranial magnetic stimulation, deep brain stimulation and transcutaneous auricular vague nerve stimulation. In the coming years, the researches trended to explore neuromodulation and mechanisms of consciousness recovery, and the main neuromodulation techniques might be deep brain stimulation and transcutaneous auricular vague nerve stimulation. ConclusionThe researches about treatment of pDOC are increasing, mainly focusing on neuromodulation, prognostic evaluation, nursing care, and training for swallowing function. More researches would focus on neuromodulation and mechanisms for restoring consciousness.

Endogenously eliminating the hematoma is a favorable strategy in addressing intracerebral hemorrhage (ICH). This study sought to determine the role of retinoid X receptor-α (RXR-α) in the context of hematoma absorption after ICH. Our results showed that pharmacologically activating RXR-α with bexarotene significantly accelerated hematoma clearance and alleviated neurological dysfunction after ICH. RXR-α was expressed in microglia/macrophages, neurons, and astrocytes. Mechanistically, bexarotene promoted the nuclear translocation of RXR-α and PPAR-γ, as well as reducing neuroinflammation by modulating microglia/macrophage reprograming from the M1 into the M2 phenotype. Furthermore, all the beneficial effects of RXR-α in ICH were reversed by the PPAR-γ inhibitor GW9662. In conclusion, the pharmacological activation of RXR-α confers robust neuroprotection against ICH by accelerating hematoma clearance and repolarizing microglia/macrophages towards the M2 phenotype through PPAR-γ-related mechanisms. Our data support the notion that RXR-α might be a promising therapeutic target for ICH.

Endogenously eliminating the hematoma is a favorable strategy in addressing intracerebral hemorrhage (ICH). This study sought to determine the role of retinoid X receptor-α (RXR-α) in the context of hematoma absorption after ICH. Our results showed that pharmacologically activating RXR-α with bexarotene significantly accelerated hematoma clearance and alleviated neurological dysfunction after ICH. RXR-α was expressed in microglia/macrophages, neurons, and astrocytes. Mechanistically, bexarotene promoted the nuclear translocation of RXR-α and PPAR-γ, as well as reducing neuroinflammation by modulating microglia/macrophage reprograming from the M1 into the M2 phenotype. Furthermore, all the beneficial effects of RXR-α in ICH were reversed by the PPAR-γ inhibitor GW9662. In conclusion, the pharmacological activation of RXR-α confers robust neuroprotection against ICH by accelerating hematoma clearance and repolarizing microglia/macrophages towards the M2 phenotype through PPAR-γ-related mechanisms. Our data support the notion that RXR-α might be a promising therapeutic target for ICH.
Anilides/pharmacology* ; Cerebral Hemorrhage/drug therapy* ; Hematoma/drug therapy* ; Humans ; Macrophages ; Microglia ; Neuroprotection ; PPAR gamma ; Retinoid X Receptor alpha