Objective:To screen differentially expressed long non-coding RNA (lncRNA) in exosomes of the brain extracellular space in mice with traumatic brain injury (TBI) and explore their biological roles in brain injury repair.Methods:Twenty-four C57BL/6J male mice were randomly divided into TBI group and control group with 12 in each group. The TBI group was treated with drill craniostomy followed by hydraulic impact while the control group was treated with drill craniostomy only. The brain tissue samples were collected at 3 hours after TBI. The exosomes from the brain extracellular space were isolated with papain digestion combined with an exosome isolation kit, the appearance of the exosomes was observed by phosphotungstic acid negative staining under a transmission electron microscope, and western blot was used to identify the exosomal marker proteins cluster of differentiation 63 (CD63), tumor susceptibility gene 101 (TSG101) and heat shock protein 70 (HSP70). After exosome identification, RNA libraries were constructed and subjected to library quality control, whole transcriptome sequencing was performed to detect, characterize and annotate lncRNA. Subsequently, the RNA high-throughput sequencing results were verified by randomly selecting five differentially expressed lncRNA. Moreover, the accuracy of the RNA high-throughput sequencing results was verified by qRT-PCR. The differentially expressed lncRNA were screened afterwards. Finally, bioinformatics analysis of differentially expressed lncRNA was performed, i.e., gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis were performed based on differentially expressed lncRNA-related genes.Results:(1) Identification of exosomes: the exosomes were cup-shaped with clear and intact membranes and diameters of 30-100 nm. Western blot confirmed the presence of typical exosome markers of the exosomes, such as CD63, TSG101, and HSP70, as well as astrocyte marker glial fibrillary acidic protein (GFAP), all of which were found in the brain tissue. (2) Validation of lncRNA high-throughput sequencing data accuracy: the 4/5 sequencing accuracy indicated that the sequencing results of lncRNAs were largely reliable. (3) Screening of differentially expressed exosomal lncRNAs after TBI: compared with the control group, a total of 442 lncRNA were significantly differentially expressed in the TBI group, including 255 up-regulated and 187 down-regulated lncRNA. (4) Bioinformatics analysis of differentially expressed lncRNAs after TBI: GO analysis showed that the differentially expressed lncRNA mainly involved in cell localization, cognition, learning or memory, were closely related to the composition of the nucleus and organelles and the binding processes of proteins, oxygen, and kinases. KEGG pathway analysis showed that upregulated lncRNA were mainly enriched in the mitogen activated protein kinase (MAPK) signaling pathway, while downregulated lncRNA were mainly enriched in the endocytosis pathway. The pathway relation network showed that MAPK signaling pathway played a key role in downstream regulation.Conclusions:There are significant differences in lncRNA expression profiles of the exosomes in the extracellular spaces of the brain cells in mice with TBI, and the differentially expressed lncRNA may mediate the process of neuronal repair, body prognosis and learning ability recovery after TBI by participating in MAPK and endocytosis signaling pathways.

Objective: To construct a hypertension risk prediction model for residents aged 18-79 years, so as to provide an assessment tool for early screening and prevention of hypertension in high-risk groups. Methods: The permanent residents aged 18-79 years from 6 townships (streets) in Fangshan District of Beijing Municipality were selected as the study subjects using a multi-stage stratified random sampling method from March to June 2023. Demographic information, lifestyle, body mass index (BMI), blood pressure, fasting blood glucose, and blood lipid were collected through questionnaire survey, physical examination, and laboratory tests. Subjects were randomly divided into training and validation sets at a 7∶3 ratio. The logistic regression model was used to screen the risk factors of hypertension, and a hypertension risk prediction nomogram was constructed. Receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis were used to verify the discrimination, fit, and clinical application value of the model. Results: A total of 4 438 subjects were included, including 2 365 males (53.29%) and 2 073 females (46.71%), with a mean age of (44.99±14.90) years. The prevalence of hypertension was 35.29% (1 566 cases), and the standardized prevalence was 24.74%. The logistic regression model screened out 9 influencing factors of hypertension. The nomogram was established as ln[p/ (1-p)]= -2.873 + 0.935×40-<50 years + 1.463×50-<60 years + 1.908×60-<70 years + 2.346×70-79 years + 0.298×male-0.675×college degree or above + 0.384×smoking + 0.227×drinking + 0.572×overweight + 1.449×obesity + 0.557×heart rate ≥80 beats/min + 0.428×diabetes + 0.484×dyslipidemia. The area under the ROC curve of the validation set was 0.821 (95%CI: 0.798-0.843), and the calibration curve results showed that the calibration curve fitted the actual curve well. Decision curve analysis showed that the threshold probability was in the range of 0.10 to 0.70, and the model had good predictive value and clinical application value. Conclusion The nomogram based on age, gender, educational level, smoking, drinking, body mass index, heart rate, diabetes, and dyslipidemia can be used to predict the risk of hypertension among residents aged 18-79 years.

Objective:To screen differentially expressed long non-coding RNA (lncRNA) in exosomes of the brain extracellular space in mice with traumatic brain injury (TBI) and explore their biological roles in brain injury repair.Methods:Twenty-four C57BL/6J male mice were randomly divided into TBI group and control group with 12 in each group. The TBI group was treated with drill craniostomy followed by hydraulic impact while the control group was treated with drill craniostomy only. The brain tissue samples were collected at 3 hours after TBI. The exosomes from the brain extracellular space were isolated with papain digestion combined with an exosome isolation kit, the appearance of the exosomes was observed by phosphotungstic acid negative staining under a transmission electron microscope, and western blot was used to identify the exosomal marker proteins cluster of differentiation 63 (CD63), tumor susceptibility gene 101 (TSG101) and heat shock protein 70 (HSP70). After exosome identification, RNA libraries were constructed and subjected to library quality control, whole transcriptome sequencing was performed to detect, characterize and annotate lncRNA. Subsequently, the RNA high-throughput sequencing results were verified by randomly selecting five differentially expressed lncRNA. Moreover, the accuracy of the RNA high-throughput sequencing results was verified by qRT-PCR. The differentially expressed lncRNA were screened afterwards. Finally, bioinformatics analysis of differentially expressed lncRNA was performed, i.e., gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis were performed based on differentially expressed lncRNA-related genes.Results:(1) Identification of exosomes: the exosomes were cup-shaped with clear and intact membranes and diameters of 30-100 nm. Western blot confirmed the presence of typical exosome markers of the exosomes, such as CD63, TSG101, and HSP70, as well as astrocyte marker glial fibrillary acidic protein (GFAP), all of which were found in the brain tissue. (2) Validation of lncRNA high-throughput sequencing data accuracy: the 4/5 sequencing accuracy indicated that the sequencing results of lncRNAs were largely reliable. (3) Screening of differentially expressed exosomal lncRNAs after TBI: compared with the control group, a total of 442 lncRNA were significantly differentially expressed in the TBI group, including 255 up-regulated and 187 down-regulated lncRNA. (4) Bioinformatics analysis of differentially expressed lncRNAs after TBI: GO analysis showed that the differentially expressed lncRNA mainly involved in cell localization, cognition, learning or memory, were closely related to the composition of the nucleus and organelles and the binding processes of proteins, oxygen, and kinases. KEGG pathway analysis showed that upregulated lncRNA were mainly enriched in the mitogen activated protein kinase (MAPK) signaling pathway, while downregulated lncRNA were mainly enriched in the endocytosis pathway. The pathway relation network showed that MAPK signaling pathway played a key role in downstream regulation.Conclusions:There are significant differences in lncRNA expression profiles of the exosomes in the extracellular spaces of the brain cells in mice with TBI, and the differentially expressed lncRNA may mediate the process of neuronal repair, body prognosis and learning ability recovery after TBI by participating in MAPK and endocytosis signaling pathways.

Objective: To investigate the prevalence, awareness, treatment and control rates of diabetes among residents in Fangshan District, Bejing Municipality, so as to provide reference for strengthening the prevention and control of diabetes. Methods: The residents aged 18 to 79 years were selected in Fangshan District using multistage stratified cluster sampling method from March to June 2023. Demographic information, lifestyle, body mass index, fasting blood glucose, and blood lipid were collected through questionnaire surveys, physical examinations and laboratory tests. The prevalence, awareness, treatment and control rates of diabetes among residents were analyzed and their influencing factors were identified using a multivariable logistic regression model. Results: A total of 4 438 residents were enrolled, including 2 367 males (53.33%) and 2 071 females (46.67%), and had a mean age of (44.99±14.90) years. The prevalence, awareness, treatment and control rates of diabetes were 15.75%, 57.37%, 56.22% and 22.90%, respectively. Multivariable logistic regression analysis showed that age (aged 30 to 39 years, OR=1.624, 95%CI: 1.011-2.609; aged 40 to 49 years, OR=3.469, 95%CI: 2.163-5.561; aged 50 to 59 years, OR=4.202, 95%CI: 2.595-6.803; aged 60 to 69 years, OR=7.229, 95%CI: 4.406-11.862; aged 70 to 79 years, OR=7.929, 95%CI: 4.665-13.479), obesity (OR=1.368, 95%CI: 1.071-1.748), hypertension (OR=1.943, 95%CI: 1.601-2.359) and dyslipidemia (OR=2.754, 95%CI: 2.287-3.317) were the factors affecting the prevalence of diabetes. Sex (women, OR=1.529, 95%CI: 1.019-2.293), age (aged 40 to 49 years, OR=4.971, 95%CI: 1.681-14.698; aged 50 to 59 years, OR=6.070, 95%CI: 2.045-18.020; aged 60 to 69 years, OR=6.902, 95%CI: 2.287-20.832; aged 70 to 79 years, OR=6.954, 95%CI: 2.191-22.070) and hypertension (OR=1.906, 95%CI: 1.337-2.718) were the factors affecting the awareness of diabetes. Age (aged 40 to 49 years, OR=4.699, 95%CI: 1.594-13.855; aged 50 to 59 years, OR=5.990, 95%CI: 2.023-17.732; aged 60 to 69 years, OR=7.134, 95%CI: 2.370-21.470; aged 70 to 79 years, OR=6.639, 95%CI: 2.102-20.964) and hypertension (OR=1.857, 95%CI: 1.304-2.645) were the factors affecting the treatment of diabetes. Age (aged 40 to 49 years, OR=2.948, 95%CI: 1.349-6.438; aged 60 to 69 years, OR=2.344, 95%CI: 1.049-5.234) was the factor affecting the control of diabetes. Conclusions The prevalence rate of diabetes among residents in Fangshan District was high, and the improvements of awareness rate, treatment rate and control rate were not obvious. Health education and management of diabetes should be strengthened for elderly, hypertensive and dyslipidemic residents.

Objective:Radiotherapy is the primary treatment for nasopharyngeal carcinoma,but it frequently leads to radiotherapy-induced temporal lobe injury(RTLI).Magnetic resonance imaging(MRI)is the main diagnostic method for RTLI after radiotherapy for nasopharyngeal carcinoma,but it is prone to missed diagnoses.This study aims to investigate the causes of missed diagnoses of RTLI in nasopharyngeal carcinoma patients undergoing MRI after radiotherapy. Methods:Clinical and MRI data from nasopharyngeal carcinoma patients diagnosed and treated with radiotherapy at Xiangya Hospital of Central South University,from January 2010 to April 2021,were collected.Two radiologists reviewed all head and neck MRIs(including nasopharyngeal and brain MRIs)before and after radiotherapy of identify cases of late delayed response-type RTLI for the first time.If the original diagnosis of the initial RTLI in nasopharyngeal carcinoma patients did not report temporal lobe lesions,it was defined as a missed diagnosis.The first diagnosis of RTLI cases was divided into a missed diagnosis group and a non-missed diagnosis group.Clinical and imaging data were compared between the 2 groups,and multivariate logistic regression analysis was used to identify independent risk factors for MRI missed diagnoses of initial RTLI. Results:A total of 187 nasopharyngeal carcinoma with post-radiotherapy RTLI were included.The original diagnostic reports missed 120 cases and accurately diagnosed 67 cases,with an initial RTLI diagnosis accuracy rate of 35.8%and a missed diagnosis rate of 64.2%.There were statistically significant differences between the missed diagnosis group and the non-missed diagnosis group in terms of lesion size,location,presence of contralateral temporal lobe lesions,white matter high signal,cystic degeneration,hemorrhage,fluid attenuated inversion recovery(FLAIR),and examination site(all P<0.05).Multivariate logistic regression analysis showed that lesions≤25 mm,non-enhancing lesions,lesions without cystic degeneration or hemorrhage,lesions located only in the medial temporal lobe,and MRI examination only of the nasopharynx were independent risk factors for missed MRI diagnosis of initial RTLI(all P<0.05). Conclusion:The missed diagnosis of initial RTLI on MRI is mainly related to lesion size and location,imaging characteristics,and MRI examination site.

Objective To investigate the expression of inositol polyphosphate 4-phosphatase type Ⅱ B(INPP4B)in colorectal cancer(CRC)and the relevant clinical significance,to determine the relationship between INPP4B and matrix metallopeptidase 7(MMP7)in CRC cells,and to make preliminary exploration of the effects of INPP4B on the proliferation and migration of CRC cells and mechanisms involved.Methods The TIMER2.0 and GEPIA2 databases were used to analyze the differences in INPP4B expression between cancer and para-cancerous tissues and the effects of such differences on the prognosis of CRC.The expression of INPP4B in 102 surgically resected CRC tumors was determined by immunohistochemistry(IHC),and the correlation between INPP4B and clinical pathological indicators was analyzed.In CRC cells with overexpressed/knocked-down INPP4B,the expression of INPP4B and MMP7 were examined by real time fluorogenic quantitative PCR,the protein expression of INPP4B was assessed by Western blot,cell proliferation was determined using the CellTiter 96? AQueous One assay,and cell migration and invasion were assessed using wound healing assay and real-time label-free dynamic cell analysis(RTCA).The LinkedOmics database was used to analyze signaling pathways related to INPP4B function,and the role of potential key molecules was validated at the cellular level.Results Analysis with the TIMER2.0 database and GEPIA2 database showed elevated INPP4B expression(colon adenocarcinoma[COAD]:2.30,rectal adenocarcinoma[READ]:2.33)in CRC compared to normal tissue(COAD:1.91,READ:1.89).IHC testing confirmed that INPP4B was upregulated in clinical CRC tissues and paracancerous tissues(P＜0.001).Cox regression model analysis showed that INPP4B(hazards ratio[HR]=1.457,95％confidence interval[CI]:1.003-2.115)affected the prognosis of CRC,and the Kaplan-Meier curve showed that patients with high INPP4B expression had shorter overall survival(P＜0.05).x2 test was performed to analyze the relationship between INPP4B expression and clinicopathological indexes,and it was found that high expression of INPP4B was correlated with lymph node metastasis(x2=3.997,P=0.046)and neural invasion(x2=8.511,P=0.004).In in vitro experiments,CRC cells overexpressing INPP4B showed a significantly increased cell proliferation and migration compared to the cells in the control group(P＜0.05).Analysis using the LinkedOmics database showed that INPP4B was correlated with extracellular matrix remodeling and cell migration.Pearson's correlation analysis showed that MMP7 was positively correlated with INPP4B(r=0.3782,P＜0.001).INPP4B overexpression or knockdown in vitro also led to the upregulation or the downregulation of MMP7 expression in CRC cells.Conclusion INPP4B is highly expressed in CRC tissues and significantly correlated with lymph node metastasis,neural invasion,and patient prognosis.MMP7 may mediate the role of INPP4B in promoting CRC cell migration and invasion.

Objective:To compare liver adhesive suspension method and V-shaped liver suspension of anterior abdominal wall in laparoscopic radical gastrectomy.Methods:This study retrospectively evaluated 115 patients who underwent laparoscopic radical gastrectomy from Jan 2018 to Dec 2019 at Shanxi Provincial People′s Hospital, 59 cases were under liver adhesive suspension (study group) and 56 cases under V-shaped liver suspension of anterior abdominal wall (control group). Intraoperative and postoperative conditions were observed and compared between the two groups.Results:All the 115 cases successfully completed surgery. The liver suspension time in study group was (2.4±0.5)min, which was significantly shorter than that of the control group((12.5±2.6) min)( t=-28.100, P<0.01). The total operation time in was (284±69) min and (307±78) min, respectively, ( t=-1.656, P=0.10). There were no statistically significant differences between the two groups in preoperative ALT, AST, total bilirubin, intraoperative incidence of hepatic lacerations and subcapsular hematoma, postoperative total bilirubin level, AST on the day 7 postoperatively, tumor location, TNM staging, surgical method, postoperative duration of hospital stay, and VAS score ( P>0.05). Conclusions:Both methods of liver suspension are safe and feasible in laparoscopic surgery for gastric cancer.Compared with V-shaped liver suspension of anterior abdominal wall, liver adhesive suspension has the advantages of less invasive, safe, and time-saving.

Purpose: This study was performed to introduce an in vivo hybrid multimodality technique involving the coregistration of micro-computed tomography (micro-CT) and high-resolution magnetic resonance imaging (HR-MRI) to concomitantly visualize and quantify mineralization and vascularization at follow-up in a rat model. Materials and Methods: Three adult female rats were randomly assigned as test subjects, with 1 rat serving as a control subject. For 20 weeks, the test rats received a weekly intravenous injection of 30 μg/kg zoledronic acid, and the control rat was administered a similar dose of normal saline. Bilateral extraction of the lower first and second molarswas performed after 10 weeks. All rats were scanned once every 4 weeks with both micro-CT and HR-MRI. Micro-CT and HR-MRI images were registered and fused in the same 3-dimensional region to quantify blood flow velocity and trabecular bone thickness at T0 (baseline), T4 (4 weeks), T8 (8 weeks), T12 (12 weeks), T16 (16 weeks), and T20 (20 weeks). Histological assessment was the gold standard with which the findings were compared. Results: The histomorphometric images at T20 aligned with the HR-MRI findings, with both test and control rats demonstrating reduced trabecular bone vasculature and blood vessel density. The micro-CT findings were also consistent with the histomorphometric changes, which revealed that the test rats had thicker trabecular bone and smaller marrow spaces than the control rat. Conclusion The combination of micro-CT and HR-MRI may be considered a powerful non-invasive novel technique for the longitudinal quantification of localized mineralization and vascularization.

OBJECTIVE: To investigate the effects of millimeter wave (MMW) exposure on apoptosis of human melanoma A375 cells and explore the mechanisms. METHODS: Through electromagnetic field calculation we simulated MMW exposure in cells and calculated the specific absorption rate (SAR). The optimal irradiation parameters were determined according to the uniformity and intensity of the SAR. A375 cells were then exposed to MMV for 15, 30, 60, or 90 min, with or without pretreatment with the caspase-3 inhibitor AC-DEVD-fmk (10 μmol/L) for 1 h at 90 min before the exposure. CCK-8 assay was used to assess the changes in the viability and Annexin-V/ PI staining was used to detect the apoptosis of the cells following the exposures; Western blotting was used to detect the expression of caspase-3 in the cells. RESULTS: The results of electromagnetic field calculation showed that for optimal MMV exposure, the incident field needed to be perpendicular to the bottom of the plastic Petri dish with the antenna placed below the dish. CCk-8 assay showed that MMW exposure significantly inhibited the cell viability in a time-dependent manner ( < 0.05); exposures for 15, 30, 60, and 90 min all resulted in significantly increased apoptosis of the cells ( < 0.05). The cells with MMW exposure showed significantly increased expression of caspase-3. The inhibitory effect of MMW on the cell viability was antagonized significantly by pretreatment of the cells with AC-DEVD-fmk ( < 0.05), which increased the cell viability rate from (36.7±0.09)% to (59.8±0.06)% ( < 0.05). CONCLUSIONS 35.2 GHz millimeter wave irradiation induces apoptosis in A375 cells by activating the caspase-3 protein.
Apoptosis ; Caspase 3 ; metabolism ; Caspase Inhibitors ; pharmacology ; Cell Line, Tumor ; Cell Survival ; Electromagnetic Fields ; Enzyme Activation ; Humans ; Magnetic Field Therapy ; Melanoma ; enzymology ; pathology ; therapy ; Time Factors