METTL3-mediated m6A modification of SOX4 regulates osteoblast proliferation and differentiation via YTHDF3 recognition.

N6-methyladenosine (m6A), the most prevalent internal modification in mRNA, is related to the pathogenesis of osteoporosis (OP). Although methyltransferase Like-3 (METTL3), an m6A transferase, has been shown to mitigate OP progression, the mechanisms of METTL3-mediated m6A modification in osteoblast function remain unclear. Here, fluid shear stress (FSS) induced osteoblast proliferation and differentiation, resulting in elevated levels of METTL3 expression and m6A modification. Through Methylated RNA Immunoprecipitation Sequencing (MeRIP-seq) and Transcriptomic RNA Sequencing (RNA-seq), SRY (Sex Determining Region Y)-box 4 (SOX4) was screened as a target of METTL3, whose m6A-modified coding sequence (CDS) regions exhibited binding affinity towards METTL3. Further functional experiments demonstrated that knockdown of METTL3 and SOX4 hampered osteogenesis, and METTL3 knockdown compromised SOX4 mRNA stability. Via RNA immunoprecipitation (RIP) assays, we further confirmed the direct interaction between METTL3 and SOX4. YTH N6-Methyladenosine RNA Binding Protein 3 (YTHDF3) was identified as the m6A reader responsible for modulating SOX4 mRNA and protein levels by affecting its degradation. Furthermore, in vivo experiments demonstrated that bone loss in an ovariectomized (OVX) mouse model was reversed through the overexpression of SOX4 mediated by adeno-associated virus serotype 2 (AAV2). In conclusion, our research demonstrates that METTL3-mediated m6A modification of SOX4 plays a crucial role in regulating osteoblast proliferation and differentiation through its recognition by YTHDF3. Our research confirms METTL3-m6A-SOX4-YTHDF3 as an essential axis and potential mechanism in OP.

Screening and identification of potential hub genes and immune cell infiltration in the synovial tissue of rheumatoid arthritis by bioinformatic approach.

Background: Rheumatoid arthritis (RA) is an autoimmune disease that affects individuals of all ages. The basic pathological manifestations are synovial inflammation, pannus formation, and erosion of articular cartilage, bone destruction will eventually lead to joint deformities and loss of function. However, the specific molecular mechanisms of synovitis tissue in RA are still unclear. Therefore, this study aimed to screen and explore the potential hub genes and immune cell infiltration in RA. Methods: Three microarray datasets (GSE12021, GSE55457, and GSE55235), from the Gene Expression Omnibus (GEO) database, have been analyzed to explore the potential hub genes and immune cell infiltration in RA. First, the LIMMA package was used to screen the differentially expression genes (DEGs) after removing the batch effect. Then the clusterProfiler package was used to perform functional enrichment analyses. Second, through weighted coexpression network analysis (WGCNA), the key module was identified in the coexpression network of the gene set. Third, the protein-protein interaction (PPI) network was constructed through STRING website and the module analysis was performed using Cytoscape software. Fourth, the CIBERSORT and ssGSEA algorithm were used to analyze the immune status of RA and healthy synovial tissue, and the associations between immune cell infiltration and RA-related diagnostic biomarkers were evaluated. Fifth, we used the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) to validate the expression levels of the hub genes, and ROC curve analysis of hub genes for discriminating between RA and healthy tissue. Finally, the gene-drug interaction network was constructed using DrugCentral database, and identification of drug molecules based on hub genes using the Drug Signature Database (DSigDB) by Enrichr. Results: A total of 679 DEGs were identified, containing 270 downregulated genes and 409 upregulated genes. DEGs were primarily enriched in immune response and chemokine signaling pathways, according to functional enrichment analysis of DEGs. WGCNA explored the co-expression network of the gene set and identified key modules, the blue module was selected as the key module associated with RA. Seven hub genes are identified when PPI network and WGCNA core modules are intersected. Immune infiltration analysis using CIBERSORT and ssGSEA algorithms revealed that multiple types of immune infiltration were found to be upregulated in RA tissue compared to normal tissue. Furthermore, the levels of 7 hub genes were closely related to the relative proportions of multiple immune cells in RA. The results of the qRT-PCR demonstrated that the relative expression levels of 6 hub genes (CD27, LCK, CD2, GZMB, IL7R, and IL2RG) were up-regulated in RA synovial tissue, compared with normal tissue. Simultaneously, ROC curves indicated that the above 6 hub genes had strong biomarker potential for RA (AUC >0.8). Conclusions: Through bioinformatics analysis and qRT-PCR experiment, our study ultimately discovered 6 hub genes (CD27, LCK, CD2, GZMB, IL7R, and IL2RG) that closely related to RA. These findings may provide valuable direction for future RA clinical diagnosis, treatment, and associated research.

Application of Triggered EMG in the Intraoperative Neurophysiological Monitoring of Posterior Percutaneous Endoscopic Cervical Discectomy.

OBJECTIVE: To describe the rationale and application of triggered EMG (T-EMG) in intraoperative neurophysiological monitoring, and to explore the efficacy and safety of posterior percutaneous endoscopic cervical discectomy (PPECD) in the treatment of cervical spondylotic radiculopathy (CSR) under multimodal intraoperative neurophysiological monitoring (IOM). METHODS: This study was a retrospective cohort control study. The clinical data of 74 patients with single-segment CSR from June 2015 to August 2018 were analyzed retrospectively, of whom 35 underwent IOM-assisted PPECD with triggered EMG (T-EMG group), while 39 were subjected to IOM-assisted PPECD alone (IOM group). Operation time, hospital stay, and complications were recorded for both groups. The curative effect was evaluated according to the Visual Analog Scale (VAS) of neck and arm pain, Japanese Orthopaedic Association (JOA) score, and modified MacNab scale. RESULTS: Operations were successful and all patients were followed up for at least 24 (average 31.77 ± 9.51) months with no patient lost to follow-up. No significant difference was found in preoperative baseline data between the T-EMG and the IOM group (P > 0.05). Also, no significant difference was found in the operation time between the T-EMG (108.29 ± 11.44 min) and the IOM (110.13 ± 12.70 min) (P > 0.05) group, but the difference in hospital stay (T-EMG: 5.66 ± 0.99 days; IOM: 7.10 ± 1.43 days) was statistically significant (P < 0.05). The VAS for the neck and upper limbs in the two groups at 1 month post-operation (T-EMG: 2.09 ± 1.07, 2.26 ± 0.92; IOM:2.18 ± 1.05, 2.31 ± 0.77) and the last follow-up (T-EMG: 0.83 ± 0.62, 0.86 ± 0.55; IOM: 0.90 ± 0.50, 0.87 ± 0.61) were significantly different from the preoperative scores (T-EMG: 6.14 ± 1.09, 7.17 ± 1.04; IOM: 6.18 ± 1.28, 7.15 ± 1.23) (P < 0.05). However, no significant difference was found between the two groups (P > 0.05). The 1-month postoperative JOA scores for the two groups (12.69 ± 0.76; 12.59 ± 0.82) and those at the last follow-up (14.60 ± 0.77; 14.36 ± 0.78) were significantly different from the preoperative scores (11.09 ± 0.98; 11.05 ± 0.89) (P < 0.05), but the difference between the two groups was not significant (P > 0.05). One patient in the T-EMG group developed a transient aggravation of symptoms on the first day after surgery. In the IOM group, three patients had intraoperative cerebrospinal fluid leakage, and symptoms of C5 nerve root paralysis were presented in four patients following surgery. Compared with the IOM group, the T-EMG group had fewer complications (1/35; 7/39, P < 0.05). At the last follow-up, the modified MacNab criteria were 91.43% (32/35) and 89.7% (35/39) for the T-EMG group and IOM group, respectively. CONCLUSIONS: Triggered EMG prevents the occurrence of neurological complications, which not only aids PPECD for CSR treatment in achieving satisfactory results, but also reduces average hospital stay and complication rates.

Percutaneous Transforaminal Endoscopic Discectomy and Fenestration Discectomy to Treat Posterior Ring Apophyseal Fractures: A Retrospective Cohort Study.

OBJECTIVE: To compare the efficacy and safety of percutaneous transforaminal endoscopic discectomy (PTED) and fenestration discectomy (FD) for posterior ring apophyseal fractures (PRAF). METHODS: This study was a retrospective cohort control study. A total of 96 patients with lumbar PRAF who underwent surgical treatment at the Henan Provincial People's Hospital of Henan University from September 2013 to December 2017 were retrospectively examined, of which 51 were treated by PTED and 45 by FD. The average age of those in the PTED group was 28.24 years, including 38 males and 13 females. The average age of those in the FD group was 28.07 years, with 33 males and 12 females. Operation time, total blood loss, hospitalization days, preoperative and postoperative visual analog scale (VAS), and Oswestry disability index (ODI) scores were evaluated. Modified MacNab criteria were used to evaluate the clinical effect at the last follow-up. RESULTS: Both operations were successful and no serious complications occurred. All patients were followed up for 12-30 (average 16.7 ± 3.2) months, and no patients were lost to follow-up. No statistically significant difference was found in the mean age and gender between the PTED group and the FD group (P < 0.05). Operation time, total blood loss, and length of hospital stay were significantly lower in the PTED group (87.65 ± 13.15 min, 12.78 ± 4.95 mL, and 6.80 ± 1.67 days, respectively) than in the FD group (114.11 ± 14.39 min, 30.89 ± 7.09 mL, and 11.71 ± 1.98 days, respectively) (P < 0.05). The VAS and ODI scores of the two groups at postoperative day 1 (PTED: 3.82 ± 0.97, 37.73% ± 3.72%; FD: 3.62 ± 1.09, 36.62% ± 3.05%), and at 3 months (PTED: 2.90 ± 1.08, 26.02% ± 2.90%; FD: 3.07 ± 0.99, 27.16% ± 4.02%), 6 months (PTED: 2.31 ± 0.88, 22.53% ± 2.67%; FD: 2.36 ± 0.77, 21.18% ± 3.35%), and the last follow-up (PTED: 1.90 ± 0.83, 19.88% ± 3.01%; FD: 1.89 ± 0.86, 18.22% ± 3.03%) were significantly different from the preoperative scores (PTED: 6.53 ± 1.00, 55.24% ± 4.54%; FD: 6.78 ± 1.31, 53.56% ± 5.73%) (P < 0.05). The VAS and ODI scores at 3 months postoperatively, 6 months postoperatively, and the last follow up were not significantly different between the two groups (P > 0.05). In the PTED group, 2 patients developed a transient nerve stimulation symptom within 1 day after surgery and 1 patient had recurrence at 3 months after surgery. In the FD group, 2 patients had severe dural ruptures due to adhesion during surgery, 1 patient developed infection complications, and 2 patients relapsed at 2 and 3 months after surgery. At the last follow-up, the modified MacNab criteria for clinical effect were 93.3% and 94.1% in the FD and PTED groups, respectively. CONCLUSION: While PTED has the same efficacy as FD for treating PRAF, it is associated with shorter operation time, less trauma, and quicker recovery.

[Spatiotemporal variation of vegetation net primary productivity and its driving factors from 2000 to 2015 in Qinling-Daba Mountains, China.] / 2000­2015å¹´ç§¦å·´å±±åŒºæ¤è¢«å‡€åˆçº§ç”Ÿäº§åŠ›æ—¶ç©ºå˜åŒ–åŠå ¶è¶‹åŠ¨å› å­.

We analyzed spatiotemporal changes in vegetation net primary productivity (NPP) in the Qinling-Daba Mountains based on MOD17A3 data and driving factor data from 2000 to 2015. We used trend analysis, Hurst index, and correlation analysis to examine the effects of climate change, soil type, topographic factors, vegetation type, and human activities on NPP. With respect to spatial variation, NPP was the highest in southwestern area and lowest in the northeast. The temporal trend over the 16-year period showed that NPP was increased in the northwest and decreased in the northeast. The projected trend of NPP was of continuity in northern area and anti-continuity in southern area of the Qinling-Daba Mountains. NPP was positively correlated with precipitation and temperature. NPP in areas with dark-brown earths, yellow earths, purplish soils and paddy soils were significantly higher than those with other soil types. The spatial distribution and trends of NPP differed among vegetation types. High NPP mainly occurred on slopes of 25° to 50° and altitudes of 500 to 1000 m or above 2500 m. Human activities had both positive and suppressive effects on NPP.

[Evaluation of the forest ecosystem service values in Qinling, China]. / 秦岭森林生态系统服务价值的时空演变.

Landsat TM images of Mt. Qinling in 1984, 2000, 2005, and 2014 were used to extract land use and land cover type by a supervised classification method in ERDAS software. Equivalent factor of ecosystem service value per unit area was revised regionally in temporal and spatial dimensions, and then was modified pixel by pixel using the vegetation coverage. Forest ecosystem service values from 1984 to 2014 were calculated with RS and GIS techniques, and the results were tested by the sensitivity index. The results showed that from 1984 to 2014, forest area accounted for 77% of the study area, and the area changes in cultivated land and construction land were the most obvious. The total ecosystem service value of Mt. Qinling increased annually and by a total of 1.68×1017 yuan during the study period. The forest ecosystem service value (its growth rate was the biggest) contributed 93.8% of the overall ecosystem service value, while water and grass ecosystem service value contributed 1.6% and 1.3%, respectively. The ecosystem service value tended to increase with increasing distance from the study area boundary, which was consistent with the spatial distribution and per-unit ecosystem service value of land use types in Mt. Qinling. The annual rate of change of each ecosystem service value was insignificant. Among the four ecosystem functions, the service value of regulation accounted for 62.7%-65.8% of the total ecosystem service value, and the service value of climate regulation showed the most dramatic increase by 4.91×1016 yuan over the study period. The sensitivity index was <1, which indicated that the modified service value index was stable and the calculated results were reliable. Overall, the forest ecosystem service value in Mt. Qinling was very large, and strengthening forest protection would be an effective measure for maintaining the stability of the Mt. Qinling ecosystem.

Role for monoaminergic systems in the antidepressant-like effect of ethanol extracts from Hemerocallis citrina.

ETHNOPHARMACOLOGICAL RELEVANCE: Hemerocallis citrina, a traditional herbal medicine, has been used for the improvement of emotions in Eastern-Asia countries. AIM OF THE STUDY: Herein, we explored the antidepressant-like effect and its monoaminergic mechanism of the ethanol extracts from Hemerocallis citrina (HCE). MATERIALS AND METHODS: Effect of HCE (90, 180 and 360 mg/kg, p.o.) on the immobility time was assessed in the mouse forced swim test (FST) and tail suspension test (TST), and locomotor activity was evaluated in the open-field test (OFT). Additionally, the monoamine neurotransmitters serotonin (5-HT), noradrenaline (NA) and dopamine (DA) levels involved in the antidepressant-like effect of HCE were also measured in the mice brain regions of frontal cortex and hippocampus. RESULTS: HCE (90, 180 and 360 mg/kg, p.o.) administration significantly reduced the immobility time in both the FST and TST without accompanying changes in locomotor activity in the OFT. The pretreatment of mice with WAY 100635 (0.1 mg/kg, s.c., a 5-HT(1A) receptor antagonist), cyproheptadine (3 mg/kg, i.p., a 5-HT(2) receptor antagonist), prazosin (62.5 µg/kg, i.p., an α(1)-adrenoceptor antagonist), yohimbine (1 mg/kg, i.p., an α(2)-adrenoceptor antagonist), propranolol (5 mg/kg, i.p., a ß-adrenoceptor antagonist) or sulpiride (50 mg/kg, i.p., a dopamine D(2) receptor antagonist), but not SCH23390 (0.05 mg/kg, s.c., a dopamine D(1) receptor antagonist) prevented the antidepressant-like effect of HCE (360 mg/kg, p.o.) in the TST. In addition, HCE enhanced 5-HT and NA levels in the frontal cortex and hippocampus as well as elevated DA levels in the frontal cortex. CONCLUSION: The results indicate that the antidepressant-like effect of HCE is dependent on the serotonergic (5-HT(1A) and 5-HT(2) receptors), noradrenergic (α(1)-, α(2)- and ß-adrenoceptors) and dopaminergic (D(2) receptor) systems as well as the elevation of 5-HT, NA and DA levels in the mouse brain.

[Forensic pathological analysis on 23 cases of fatal pulmonary thromboembolism].

OBJECTIVE: To analyze the forensic pathological characteristics of sudden death caused by pulmonary thromboembolism and the chronological transformation of thrombus and explore the assessment method of the causal relationship between previous trauma and the following fatal PTE episode. METHODS: All the 23 cases reviewed here were collected from our institute files from the year of 1998 to 2008. RESULTS: Trauma, surgery and braking etc. were all risky factors of PTE. Of these cases, 12 cases were caused by trauma, 21 cases were caused by surgery and 22 cases died in hospitals which were often happened one or two weeks after injury or one week's postoperative time. Of all the cases, 6 cases had single attack of thrombus and the rest 17 cases had the recurrence of thrombus. The number of the leg deep vein to be the embolic source was 16 cases which were often seen in the left leg. CONCLUSION: It is important to confirm the embolic source, trauma, surgery and chronological events in determing the sudden death with PTE.