Neurological diseases are a major health concern, and brain injury is a typical pathological process in various neurological disorders. Different biomarkers in the blood or the cerebrospinal fluid are associated with specific physiological and pathological processes. They are vital in identifying, diagnosing, and treating brain injuries. In this review, we described biomarkers for neuronal cell body injury (neuron-specific enolase, ubiquitin C-terminal hydrolase-L1, αII-spectrin), axonal injury (neurofilament proteins, tau), astrocyte injury (S100β, glial fibrillary acidic protein), demyelination (myelin basic protein), autoantibodies, and other emerging biomarkers (extracellular vesicles, microRNAs). We aimed to summarize the applications of these biomarkers and their related interests and limits in the diagnosis and prognosis for neurological diseases, including traumatic brain injury, status epilepticus, stroke, Alzheimer's disease, and infection. In addition, a reasonable outlook for brain injury biomarkers as ideal detection tools for neurological diseases is presented.
Humans ; Biomarkers/cerebrospinal fluid* ; Nervous System Diseases/diagnosis* ; Brain Injuries/metabolism* ; Phosphopyruvate Hydratase/cerebrospinal fluid* ; Glial Fibrillary Acidic Protein/blood* ; S100 Calcium Binding Protein beta Subunit/blood* ; tau Proteins/cerebrospinal fluid* ; Ubiquitin Thiolesterase/blood* ; Myelin Basic Protein/cerebrospinal fluid* ; Neurofilament Proteins/blood* ; MicroRNAs/blood* ; Brain Injuries, Traumatic/metabolism*

Objective To investigate whether PLCB1 expression leads to gastric adenocarcinoma metastasis and poor prognosis, and to preliminarily analyze its mechanism. Methods 122 gastric adenocarcinoma patients and their adjacent non-cancerous tissues were selected, and tissue microarray technology was used to detect the expression levels of PLCB1, epithelial cadherin(E-cadherin), vimentin and CD8⁺ T cells by immunohistochemistry, and scored by two pathologists. According to the immunohistochemical score of PLCB1, the patients were divided into PLCB1 high expression group (IHC>90) and PLCB1 low expression group (IHC≤90). The clinical pathological characteristics, epithelial mesenchymal transition(EMT)-related proteins and CD8⁺ T cells expression differences between the two groups were compared. The overall survival of the patients was collected, and COX regression analysis and Kaplan-Meier curve were used to evaluate the relationship between PLCB1 expression level and prognosis. Results PLCB1 was highly expressed in 55 cases of gastric adenocarcinoma tissues, while only 12 cases in adjacent non-cancerous tissues. The tumor invasion depth, lymph node metastasis degree and TNM stage of the PLCB1 high expression group were higher than those of the PLCB1 low expression group. Chi-square test showed that PLCB1 expression level was negatively correlated with E-cadherin (r=-0.339), positively correlated with vimentin (r=0.211), and negatively correlated with CD8⁺ T cells (r=-0.343). Kaplan-Meier curve analysis showed that the overall survival and disease-free survival of gastric adenocarcinoma patients with high PLCB1 expression were significantly reduced. Multivariate COX regression analysis showed that except for lymph node metastasis, tumor invasion depth, TNM stage, E-cadherin and vimentin were also independent prognostic factors for gastric adenocarcinoma patients. Conclusion PLCB1 is highly expressed in gastric adenocarcinoma, and is closely related to tumor aggressiveness and prognosis. PLCB1 may induce EMT and inhibit CD8⁺ T cell infiltration to affect gastric adenocarcinoma metastasis and immune response.
Humans ; Stomach Neoplasms/genetics* ; Epithelial-Mesenchymal Transition ; Male ; Female ; Middle Aged ; Prognosis ; Adenocarcinoma/genetics* ; Cadherins/metabolism* ; Aged ; Adult ; CD8-Positive T-Lymphocytes/immunology* ; Vimentin/metabolism* ; Lymphatic Metastasis ; Neoplasm Metastasis

Objective To explore the mechanism of TPT1-AS1 targeting miR-324/TWIST1 axis to regulate the proliferation, invasion, migration and angiogenesis of epithelial ovarian cancer (EOC) cells, thereby affecting ovarian cancer (OC) progression. Methods RT-qPCR was used to detect the expression of TPT1-AS1 and miR-324 in 29 OC lesions and adjacent tissue samples. The two OC cell models of TPT1-AS1 overexpression and miRNA324 knockdown were constructed, and the cell proliferation, invasion and migration abilities were detected by CCK-8, Transwell^TM and scratch test. Western blot analysis was used to detect the protein expression levels of TWIST1, epithelial cadherin (E-cadherin), Vimentin, and vascular endothelial growth factor A (VEGF-A) in OC cells. Fluorescence in situ hybridization (FISH) and RNA pull-down experiments were used to verify the interaction between TPT1-AS1 and miR-324. Immunohistochemistry and Targetscan bioinformatics analysis were used to verify the negative regulatory role of miR-324 in the epithelial-mesenchymal transition (EMT) process. Results The TPT1-AS1 expression was significantly higher in OC tissues than that in para-cancerous tissues, while the miR-324 expression was significantly lower. In SKOV3 cells with TPT1-AS1 overexpression, the miR-324 expression decreased significantly, and TPT1-AS1 was negatively correlated with miR-324. It was also found that TPT1-AS1 and miR-324 were co-expressed in OC cells, and there was a direct binding relationship between them. Down-regulation of miR-324 significantly promoted the proliferation, invasion and migration of SKOV3 cells. Further studies revealed that miR-324 had a binding site at the 3'-UTR end of the TWIST1, a key transcription factor for EMT. Inhibiting miR-324 expression increased the transcription level of TWIST1, leading to a decrease in E-cadherin protein expression and an increase in Vimentin protein expression. Additionally, the downregulation of miR-324 resulted in an increased expression level of VEGF-A protein, which in turn enhanced angiogenesis of OC. Conclusion TPT1-AS1 promotes EOC cell proliferation, invasion, migration and angiogenesis by negatively regulating the miR-324/TWIST1 axis, thus promoting the development of OC. These findings provide new potential targets for the diagnosis and treatment of OC.
Humans ; MicroRNAs/metabolism* ; Female ; Cell Movement/genetics* ; Ovarian Neoplasms/blood supply* ; Twist-Related Protein 1/metabolism* ; Cell Line, Tumor ; Neovascularization, Pathologic/genetics* ; Neoplasm Invasiveness ; Carcinoma, Ovarian Epithelial/metabolism* ; Nuclear Proteins/metabolism* ; Cell Proliferation/genetics* ; Epithelial-Mesenchymal Transition/genetics* ; Gene Expression Regulation, Neoplastic ; RNA, Long Noncoding/metabolism* ; Cadherins/genetics* ; Vascular Endothelial Growth Factor A/genetics* ; Vimentin/genetics* ; Angiogenesis

OBJECTIVE: To evaluate the wound healing of recipient and donor sites following keratinized mucosa augmentation (KMA) around implants in reconstructed jaw areas and to compare these outcomes with gingival grafts in native jawbone, so as to provide clinical guidance for postoperative maintenance, and to investigate the impact of clinical experience on the evaluation of KMA postoperative healing through subgroup comparisons. METHODS: This study included patients who underwent resection of maxillofacial tumors, fibular or iliac flap reconstruction, and implant placement at Peking University Dental Hospital from October 2020 to April 2023. Three months post-implant placement, the patients were referred for KMA procedures. Clinical photographs of the reconstructed area were taken preoperatively, immediately postoperatively, and 3 weeks and 3 months post-surgery. Additionally, photographs of the palatal donor site were obtained preoperatively and 3 weeks later. Wound healing was assessed by four junior and three senior clinicians utilizing the early healing index (EHI), early wound healing score (EHS), and pink esthetic score (PES).And senior clinicians evaluated the healing effect compared with gingival transplantation on natural jawbone using a 10-point scale. RESULTS: A total of 26 patients with jawbone reconstruction were included, with an average age of (34.2±10.2) years, 11 males (42.3%) and 15 females (57.7%). Among them, 13 cases (50.0%) underwent fibula flap reconstruction, and 13 cases (50.0%) underwent iliac flap reconstruction. The average number of implants per patient was 3.2±0.7. In the recipient area, 3 weeks postoperatively, the EHS was 7.0 (4.0, 9.0), with sub-item scores as follows: Clinical signs of re-epithelialization (CSR) 6.0 (3.0, 6.0), clinical signs of haemostasis (CSH) 1.5 (1.0, 2.0), and clinical signs of inflammation (CSI) 1.0 (0.0, 1.0), indicating that the average appearance of the wound in the recipient area was characterized by generally well-approximated wound edges with minimal fibrin lines and mild erythema and swelling. The EHI for the recipient area was 2.0 (1.5, 2.5), suggesting that the incision was mostly closed with some fibrin lines 3 weeks postoperatively. The long-term healing evaluation system, PES, was 2.5 (2.0, 3.0), with sub-scores for color [1.0 (1.0, 1.5)] and texture [1.5 (1.0, 2.0)], which were slightly different from the reference values.In the palatal donor area, 3 weeks postoperatively, the EHI score was lower at 1.3 (1.0, 2.5), while the EHS score was higher at 8.5 (6.0, 10.0), indicating better soft tissue healing in the donor area compared with the recipient area. Among the clinicians with different levels of experience, the assessment of wound healing revealed that except for the CSI sub-item, where the junior group scored higher than the senior group, all other sub-items showed significantly higher scores in the senior group compared with the junior group. In the EHS evaluation system, the CSH sub-item demonstrated no significant differences between the groups with varying levels of experience. Experienced clinicians' evaluation outcomes of healing effect compared with gum graft on natural alveolar bone was 8.5 (7.5, 9.5), showing high consistency [intraclass correlation coefficient (ICC): 0.892; 95% confidence interval (<i>CIi>): 0.791－0.949], suggesting slightly suboptimal healing results after KMA surgery. CONCLUSION The healing process following KMA in the context of jawbone reconstruction is relatively protracted, emphasizing the necessity for comprehensive postoperative management. Moreover, clinician experience plays a significant role in the assessment of wound healing outcomes for KMA in maxillofacial reconstruction.
Humans ; Wound Healing ; Adult ; Male ; Female ; Gingiva/transplantation* ; Plastic Surgery Procedures/methods* ; Middle Aged ; Surgical Flaps ; Keratins

OBJECTIVES: Esophageal squamous cell carcinoma (ESCC) is characterized by complex pathogenesis and poor prognosis. In recent years, epithelial-mesenchymal transition (EMT) in tumor initiation and progression has attracted increasing attention. Enhancer of zeste homolog 2 (EZH2), which is aberrantly expressed in various tumors, may be closely related to the EMT process. This study aims to examine the expression and correlation of EZH2 and EMT markers in ESCC cells and tissues, evaluate the effects of <i>EZH2i> knockdown on ESCC cell proliferation, invasion, and migration, and explore how EZH2 contributes to the malignant biological behavior of ESCC. METHODS: Bioinformatics analyses were used to assess EZH2 expression levels in ESCC. Small interfering RNA was used to knock down <i>EZH2i> in ESCC cell lines EC109 and EC9706. Cell proliferation, invasion, and migration were evaluated using cell counting kit-8 (CCK-8), wound healing, and Transwell assays. Protein and mRNA expression levels of EZH2, E-cadherin (E-cad), and vimentin (Vim) were detected by Western blotting and real time fluorogenic quantitative PCR (RT-qPCR), respectively. Immunohistochemical (IHC) staining was performed on 70 ESCC tissue samples and 40 paired adjacent normal tissues collected from the First Affiliated Hospital of Shihezi University between 2010 and 2016 to assess the expression of EZH2, E-cad, and Vim, and to analyze their associations with clinicopathological feature and patient prognosis. RESULTS: Bioinformatics analysis showed that <i>EZH2i> was highly expressed in ESCC (<i>Pi><0.001), and high <i>EZH2i> expression was associated with worse prognosis (<i>Pi><0.001). CCK-8, wound healing, and Transwell assays demonstrated that <i>EZH2i> knockdown significantly suppressed the proliferation, invasion, and migration of ESCC cells (<i>Pi><0.001). In addition, Vim expression was significantly reduced, while E-cad expression was significantly increased at both protein and mRNA levels in <i>EZH2i>-silenced cells (all <i>Pi><0.05). IHC staining analysis revealed higher expression of EZH2 and Vim and lower expression of E-cad in ESCC tissues compared to adjacent normal tissues. Kaplan-Meier survival analysis showed that low expression of <i>EZH2i> and <i>Vimi> and high expression of <i>E-cadi> were associated with longer survival (all <i>Pi><0.05). CONCLUSIONS EZH2 promotes malignant biological behavior in ESCC by mediating EMT. Elevated EZH2 expression is associated with poor prognosis in ESCC patients.
Humans ; Enhancer of Zeste Homolog 2 Protein/physiology* ; Esophageal Squamous Cell Carcinoma/pathology* ; Epithelial-Mesenchymal Transition/genetics* ; Esophageal Neoplasms/metabolism* ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement ; Cadherins/genetics* ; Vimentin/genetics* ; Male ; Female ; Middle Aged ; Neoplasm Invasiveness ; Prognosis ; RNA, Small Interfering/genetics* ; Gene Expression Regulation, Neoplastic

Objective:The mucosa of Chronic rhinosinusitis with nasal polyps（CRSwNP） is accompanied by tissue remodeling. Epithelial-mesenchymal transition（EMT） plays an important role in tissue remodeling, but the mechanism of EMT is not yet clear. The purpose of this study is to further clarify the pathogenesis of CRSwNP and provide another idea and theoretical basis for the treatment of CRSwNP. Methods:①The expression of NLRP3 and EMT-related protein（E-cadherin, Vimentin） in the nasal mucosa of the CRSwNP group and the normal control group were detected by immunohistochemistry（IHC）. ②Primary human nasal epithelial cells（HNECs） were cultured in vitro, and HNE-intervened cells with different concentrations（0, 10, 25, 50, 100 ng/mL） were used. After stimulation for 24 h, mRNA and protein expressions of E-cadherin, Vimentin, NLRP3 were detected by qRT-PCR and western blotting. ③Cells were collected at 0, 24, 36, 48 and 72 hours later after incubation with HNE with the optimal concentration, and the mRNA and protein expressions of E-cadherin, Vimentin and NLRP3 were detected by qRT-PCR and western blotting. ④Primary human nasal epithelial cells were pretreated with NLRP3 inhibitor MCC950, then stimulated with HNE, and EMT-related proteins（E-cadherin, Vimentin） and NLRP3 expression were detected by qRT-PCR and western blotting. Results:①The expression levels of NLRP3 and Vimentin in nasal polyps of CRSwNP patients were higher than those of control group, and the expression of E-cadherin was lower（<i>Pi><0.05）. The mRNA and protein expression levels of NLRP3 and Vimentin increased when HNE stimulated primary human nasal epithelial cells, while the expression of E-cadherin decreased. ②The effect was most significant when the HNE stimulated nasal mucosal epithelial cells were exposed to 50 ng/mL（<i>Pi><0.05）. The primary human nasal epithelial cells were stimulated with 50 ng/ml HNE, and the effect was most significant when the duration of HNE exposure was 36 h（<i>Pi><0.05）. ③Primary human nasal epithelial cells were pretreated with MCC950 and then stimulated with HNE. The mRNA and protein expression levels of E-cadherin in the NLRP3 inhibitor pretreated group were increased, while the mRNA and protein expression levels of Vimentin and NLRP3 were decreased（<i>Pi><0.05）. Conclusion:ln CRSwNP, HNE promotes EMT in human nasal mucosal epithelial cells by activating NLRP3.
Humans ; Nasal Polyps/metabolism* ; Epithelial-Mesenchymal Transition ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Sinusitis/metabolism* ; Cadherins/metabolism* ; Vimentin/metabolism* ; Chronic Disease ; Nasal Mucosa/cytology* ; Rhinitis/metabolism* ; Epithelial Cells/metabolism* ; Cells, Cultured ; Rhinosinusitis

Peri-implant keratinized mucosa (PIKM) augmentation refers to surgical procedures aimed at increasing the width of PIKM. Consensus reports emphasize the necessity of maintaining a minimum width of PIKM to ensure long-term peri-implant health. Currently, several surgical techniques have been validated for their effectiveness in increasing PIKM. However, the selection and application of PIKM augmentation methods may present challenges for dental practitioners due to heterogeneity in surgical techniques, variations in clinical scenarios, and anatomical differences. Therefore, clear guidelines and considerations for PIKM augmentation are needed. This expert consensus focuses on the commonly employed surgical techniques for PIKM augmentation and the factors influencing their selection at second-stage surgery. It aims to establish a standardized framework for assessing, planning, and executing PIKM augmentation procedures, with the goal of offering evidence-based guidance to enhance the predictability and success of PIKM augmentation.
Humans ; Consensus ; Dental Implants ; Mouth Mucosa/surgery* ; Keratins

Objective: To investigate the morphology and immunohistochemical (IHC) expression of pseudostratified ependymal tubules in ovarian mature teratoma (MT). Methods: Five cases of ovarian MT with pseudostratified ependymal tubules were collected from Shenzhen Hospital(Futian) of Guangzhou University of Chinese Medicine and the Eighth Affiliated Hospital of Sun Yat-sen University from March 2019 to March 2022. In addition, 15 cases of ovarian MT with monolayer ependymal epithelium from Shenzhen Hospital (Futian) of Guangzhou University of Chinese medicine and seven cases of immature teratoma (IMT) from Hainan Provincial People's Hospital from March 2019 to March 2022 were collected as control. The morphologic characteristics and immunophenotypes of pseudostratified ependymal tubules, monolayer ependymal epithelium, and primitive neural epithelial tubules were observed and compared by H&E stain and IHC expression pattern of genes related to the differentiation status of neuroepithelium, namely SALL4, Glypican3, nestin, SOX2, Foxj1, and Ki-67. Results: Mean age of the five patients of ovarian MT with pseudostratified ependymal tubules was 26 years (range from 19 to 31 years). Two tumors were located in the left ovary and three in the right. All five cases were excised, and clinical follow-up was available (mean follow-up 1.5 years; range 0.5 to 3 years). No recurrence was noted in any cases. The pseudostratified ependymal tubules of ovarian MT, which were lined with columnar or oval epithelia up to 4-6 layers, were morphologically similar to the primitive neuroepithelial tubules of IMT and different from monolayer ependymal epithelium of ovarian MT. By immunohistochemistry, SALL4 and Glypican3 were negative, Foxj1 was positive and Ki-67 index was lower in the pseudostratified ependymal tubules and the monolayer ependymal epithelium of ovarian MT. However, the primitive neuroepithelial tubules of IMT showed variably expression of SALL4 and Glypican3, were negative for Foxj1 and high Ki-67 index. All the above three groups expressed nestin and SOX2. Conclusions: The pseudostratified ependymal tubules of ovarian MT, which have morphological similarities to the primitive neuroepithelial tubules of IMT, are similar to the monolayer ependymal epithelia of the MT in immunophenotype. IHC assessment of Foxj1 and Ki-67 is helpful to differentiate the pseudostratified ependymal tubules of ovarian MT from the primitive neuroepithelial tubules of IMT.
Female ; Humans ; Young Adult ; Adult ; Nestin ; Ki-67 Antigen ; Immunohistochemistry ; Ovarian Neoplasms/pathology* ; Teratoma/pathology*

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.
Male ; Mice ; Animals ; Keratin-18 ; Actins ; Desmin ; Liver ; Hepatocytes ; Hepatic Stellate Cells

BACKGROUND: Long non-coding RNA colon cancer-associated transcript 1 (CCAT1) is involved in transforming multiple cancers into malignant cancer types. Previous studies underlining the mechanisms of the functions of CCAT1 primarily focused on its decoy for miRNAs (micro RNAs). However, the regulatory mechanism of CCAT1-protein interaction associated with tumor metastasis is still largely unknown. The present study aimed to identify proteome-wide CCAT1 partners and explored the CCAT1-protein interaction mediated tumor metastasis. METHODS: CCAT1-proteins complexes were purified and identified using RNA antisense purification coupled with the mass spectrometry (RAP-MS) method. The database for annotation, visualization, and integrated discovery and database for eukaryotic RNA binding proteins (EuRBPDB) websites were used to bioinformatic analyzing CCAT1 binding proteins. RNA pull-down and RNA immunoprecipitation were used to validate CCAT1-Vimentin interaction. Transwell assay was used to evaluate the migration and invasion abilities of HeLa cells. RESULTS: RAP-MS method worked well by culturing cells with nucleoside analog 4-thiouridine, and cross-linking was performed using 365 nm wavelength ultraviolet. There were 631 proteins identified, out of which about 60% were RNA binding proteins recorded by the EuRBPDB database. Vimentin was one of the CCAT1 binding proteins and participated in the tumor metastasis pathway. Knocked down vimetin ( VIM ) and rescued the downregulation by overexpressing CCAT1 demonstrated that CCAT1 could enhance tumor migration and invasion abilities by stabilizing Vimentin protein. CONCLUSION CCAT1 may bind with and stabilize Vimentin protein, thus enhancing cancer cell migration and invasion abilities.
Humans ; HeLa Cells ; RNA, Long Noncoding/metabolism* ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Vimentin/metabolism* ; MicroRNAs/metabolism* ; Colonic Neoplasms/genetics* ; RNA-Binding Proteins/metabolism* ; Gene Expression Regulation, Neoplastic/genetics* ; Cell Movement/genetics*