Enhancer-driven transcription of MCM8 by E2F4 promotes ATR pathway activation and glioma stem cell characteristics.

BACKGROUND: Glioma stem cells (GSCs) are responsible for glioma recurrence and drug resistance, yet the mechanisms underlying their maintenance remains unclear. This study aimed to identify enhancer-controlled genes involved in GSCs maintenance and elucidate the mechanisms underlying their regulation. METHODS: We analyzed RNA-seq data and H3K27ac ChIP-seq data from GSE119776 to identify differentially expressed genes and enhancers, respectively. Gene Ontology analysis was performed for functional enrichment. Transcription factors were predicted using the Toolkit for Cistrome Data Browser. Prognostic analysis and gene expression correlation was conducted using the Chinese Glioma Genome Atlas (CGGA) data. Two GSC cell lines, GSC-A172 and GSC-U138MG, were isolated from A172 and U138MG cell lines. qRT-PCR was used to detect gene transcription levels. ChIP-qPCR was used to detect H3K27ac of enhancers, and binding of E2F4 to target gene enhancers. Western blot was used to analyze protein levels of p-ATR and γH2AX. Sphere formation, limiting dilution and cell growth assays were used to analyze GSCs growth and self-renewal. RESULTS: We found that upregulated genes in GSCs were associated with ataxia-telangiectasia-mutated-and-Rad3-related kinase (ATR) pathway activation, and that seven enhancer-controlled genes related to ATR pathway activation (LIN9, MCM8, CEP72, POLA1, DBF4, NDE1, and CDKN2C) were identified. Expression of these genes corresponded to poor prognosis in glioma patients. E2F4 was identified as a transcription factor that regulates enhancer-controlled genes related to the ATR pathway activation, with MCM8 having the highest hazard ratio among genes positively correlated with E2F4 expression. E2F4 bound to MCM8 enhancers to promote its transcription. Overexpression of MCM8 partially restored the inhibition of GSCs self-renewal, cell growth, and the ATR pathway activation caused by E2F4 knockdown. CONCLUSION: Our study demonstrated that E2F4-mediated enhancer activation of MCM8 promotes the ATR pathway activation and GSCs characteristics. These findings offer promising targets for the development of new therapies for gliomas.

[Efficacy of percutaneous short segment fixation in the treatment of Magerl A3 thoracolumbar fractures with low bone mineral density:a retrospective study].

OBJECTIVE: To explore the clinical efficacy of percutaneous pedicle screw short segment internal fixation with or without the intermediate screw in the treatment of Magerl A3 thoracolumbar fractures with low bone mineral density. METHODS: Patients with Magerl A3 thoracolumbar fracture underwent percutaneous pedicle screw short segment internal fixation from January 2017 to July 2020 were retrospectively analyzed, 93 cases met the diagnosis and inclusion criteria, 9 cases were excluded according to the exclusion criteria, and the remaining 84 cases obtained complete imaging follow-up data. There were 38 males and 46 females, the age ranged from 56 to 73 years old with an average of (64.78±7.12) years old, bone mineral density (BMD) ranged from 0.61 to 0.89 g/cm3 with an average of (0.73±0.14) g/cm3, the follow-up time was 11 to 25 months with an average of (17.58±6.12) months. There were 45 cases in group A with intermediate screw and 39 cases in group B without intermediate screw. The operation time and intraoperative blood loss were recorded, Oswestry Disability Index (ODI) and visual analogue scale (VAS) were used for clinical evaluation. The Cobb angle, vertebral wedge angle (VWA) and anterior vertebral body height (AVBH) were measured by X-ray after the operation. The corrected loss of the above parameters was calculated. RESULTS: There were 5 cases of screw loosening in 84 patients (2 cases in group A and 3 cases in group B, P>0.05). There were significant differences in operation time and intraoperative blood loss between two groups(P<0.01). Clinical effects of two groups were good, postoperative VAS and ODI after operation obviously improved, there was no significant difference between two groups during all follow-up periods (3 days, 1 month after operation and the final follow-up) (P>0.05). Three days after the operation, the image evaluations (Cobb angle, VWA and AVBH) were significantly improved (P<0.05), but significant reduction loss was observed in both groups at 1 month after the operation and at the final follow-up (P<0.05). At the final follow-up, the loss of Cobb angle, VWA and AVBH in group A were (5.26±4.18) °, (4.63±3.80) ° and (9.54±8.71)%, respectively;group B was (6.01±4.34) °, (6.55±6.21) ° and (11.67± 9.95)%, respectively;however, there was no significant difference in reduction loss between the two groups(P>0.05). CONCLUSION: Although the curative effect of the patients is satisfactory, the stability of the patients can not be improved by increasing the middle injured vertebra screw placement, the two groups of percutaneous short segment internal fixation can not resist the reduction loss of Magerl-A3 thoracolumbar fracture with low bone mineral density. Because the injured vertebra screw increases the operation time and intraoperative blood loss, it is not significant to use the intermediate screw for the elderly Magerl A3 thoracolumbar fractures with low bone mineral density.

Long noncoding RNA OIP5-AS1 targets Wnt-7b to affect glioma progression via modulation of miR-410.

The present study was undertaken to investigate the underlying mechanisms of long noncoding RNA OIP5-AS1 via regulating miR-410 to modulate Wnt-7b in the progression of glioma. To address this problem, we measured the expression of OIP5-AS1 and miR-410 in glioma tissues by qRT-PCR. Glioma U87 cells were transfected with OIP5-AS1 siRNA or miR-410 inhibitors. The targeting relationships among miR-410, OIP5-AS1 and Wnt-7b were verified by luciferase reporter assays. Western blotting was employed to determine the expression of Wnt-7b/ß-catenin pathway-related proteins, while MTT, flow cytometry, Transwell assays and wound-healing assays were used to measure the biological characteristics of glioma cells. The results showed that OIP5-AS1 expression was higher and miR-410 was lower in glioma tissues. Luciferase reporter assays confirmed a targeting relationship between OIP5-AS1 and miR-410, as well as between miR-410 and Wnt-7b. Silencing OIP5-AS1 reduced cell proliferation, invasion and migration of glioma U87 cells and led to depressed expression levels of miR-410, Wnt-7b, p-ß-catenin, GSK-3ß-pS9, c-Myc and cyclin D1. Furthermore, down-regulation of OIP5-AS1 induced G0/G1 phase cell cycle arrest and apoptosis of glioma cells. Inhibitors of miR-410 abolished the biological effects of OIP5-AS1 siRNA in glioma cells. In vivo, OIP5-AS1 knockdown also inhibited tumor growth. Taken together, this research suggested that silencing OIP5-AS1 may specifically block the Wnt-7b/ß-catenin pathway via targeted up-regulating miR-410, thereby inhibiting growth, invasion and migration while promoting apoptosis in glioma cells.

Exosomal miR-214-5p Released from Glioblastoma Cells Modulates Inflammatory Response of Microglia after Lipopolysaccharide Stimulation through Targeting CXCR5.

BACKGROUND AND OBJECTIVE: Exosomes communicate inter-cellularly and miRNAs play critical roles in this scenario. MiR-214-5p was implicated in multiple tumors with diverse functions uncovered. However, whether miR-214-5p is mechanistically involved in glioblastoma, especially via exosomal pathway, is still elusive. Here we sought to comprehensively address the critical role of exosomal miR-214-5p in glioblastoma (GBM) microenvironment. METHODS: The relative expression of miR-214-5p was determined by real-time PCR. Cell viability and migration were measured by MTT and transwell chamber assays, respectively. The secretory cytokines were measured with ELISA kits. The regulatory effect of miR-214-5p on CXCR5 expression was interrogated by luciferase reporter assay. Protein level was analyzed by Western blot. RESULTS: We demonstrated that miR-214-5p was aberrantly overexpressed in GBM and associated with poorer clinical prognosis. High level of miR-214-5p significantly contributed to cell proliferation and migration. GBM-derived exosomal miR-214-5p promoted inflammatory response in primary microglia upon lipopolysaccharide challenge. We further identified CXCR5 as the direct target of miR-214- 5p in this setting. CONCLUSION: Overexpression of miR-214-5p in GBM modulated the inflammatory response in microglia via exosomal transfer.

PPD results after 3-6 years of BCG vaccination and its effect on latent tuberculosis infection in children / 中国学校卫生

Objective@#To understand the results of tuberculin skin test (PPD) in preschool children after the vaccination of BCG, and to analyze the effect of BCG vaccination on latent tuberculosis infection in children.@*Methods@#From January to November 2018, a total of 1 359 preschool children from 14 kindergartens in 8 districts and cities of Jiangsu Province were selected for tuberculin test(PPD), and chest X-ray examination was performed on children with strong PPD results.@*Results@#The positive rate of PPD in preschool children in Jiangsu Province was 23.33%, of which strong positive and moderate positive (PPD≥10 mm) were totaled 6.47%. There were 149 boys (21.29%) with PPD positive reactions and 168 girls(25.50%) with PPD positive reactions, and differences of PPD positive reactions with different genders were of no statistical significance (χ2=3.36, P>0.05) And there were 201 children (25.35%) with PPD positive reactions in northern Jiangsu, 116 children (20.50%) with PPD positive reactions in southern Jiangsu. There were significant differences in the results of PPD positive and negative reactions between different regions(χ2=4.35, P<0.05). There was 1 case of PPD positive reactions among 3-year-old children(0.71%), 19 cases among 4-year-old children(3.89%), 31 cases among 5-year-old children(8.96%), and 37 cases among 6-year-old children(9.63%), and the differences of PPD positive reactions of different age groups were of statistical significance(χ2=21.69, P<0.01).@*Conclusion@#The positive rate of PPD in preschool children in Jiangsu Province is very low, indicating that PPD can be used as a detection method for latent infection in children. The overall positive rate of PPD in preschool children in Jiangsu Province is also low, and appropriate measures should be taken to protect susceptible children and effectively prevent and control childhood tuberculosis.

Downregulation of ZMYND11 induced by miR-196a-5p promotes the progression and growth of GBM.

ZMYND11 (zinc finger MYND-type containing 11) has been widely regarded to be involved in a variety of cancers as a potential suppressor. However, the biological role and mechanism of ZMYND11 in glioblastoma multiform (GBM) remain unknown. In this study, we found that ZMYND11 expression was remarkably decreased in GBM tissues from 20 cases and cell line (U87) compared to normal brain tissue from 10 cases (P < 0.001). Furthermore, we explored that ZMYND11 upregulation significantly suppressed U87 cells proliferation and invasion, induced cell cycle arrest and apoptosis in vitro. Subsequently, we identified increased ZMYND11 inhibited the tumor growth using tumor cells xenograft experiment on rude mice. Moreover, we explored that ZMYND11 was a new direct and functional target of miR-196a-5p in U87 via luciferase reporter assay. In addition, we confirmed the negative correlation between miR-196a-5p and ZMYND11 in GBM tissue and U87 cells by changing the expression level of miR-196a-5p with lentivirus and plasmid vector. Furthermore, we demonstrated that decreased ZMYND11 could reverse suppressive effect of downregulated miR-196a-5p on U87 by rescue experiment. Taken together, ZMYND11 was demonstrated to be a potential and extremely promising suppressor of GBM, while miRNA-196a-5p was quite an important target of treatment of GBM.

One-Dimensional Zinc-Based Coordination Polymer as a Higher Capacity Anode Material for Lithium Ion Batteries.

A zinc-based one-dimensional (1D) coordination polymer ([Zn(H2mpca)2(tfbdc)(H2O)], Zn-ODCP) has been synthesized and characterized by spectroscopic and physicochemical methods, single-crystal X-ray diffraction, and thermogravimetric analysis (H2mpca = 3-methyl-1H-pyrazole-4-carboxylic acid; H2tfbdc = 2,3,5,6-tetrafluoroterephthalic acid). Zn-ODCP shows blue luminescence in the solid state. When Zn-ODCP acts as an anode material for lithium ion batteries, it exhibits a good cyclic stability and a higher reversible capacity of 300 mAh g-1 at 50 mA g-1 after 50 cycles. The higher capacity may be mainly ascribed to the metal ion and ligand all taking part in lithium storage. Searching for electrode materials of lithium ion batteries from 1D metal coordination polymers is a new route.

[Effect of hyacinth mulching on rice (Oryza sativa L.) uptake and utilization of nitrogen].

Nitrogen is the most important element for rice growth, and hyacinth can absorb large quantities of nitrogen and accumulate in their tissues. The field experiment was conducted to investigate the effects of hyacinth mulching on rice nitrogen (N) concentration, uptake, efficiency and allocation at different growth stages. Taked Yun 2645 as a material, the nitrogen uptake and utilization of rice were evaluated under two different levels of N input: low (LN, 120 kg x hm(-2)) and normal N (NN, 240 kg x hm(-2)) in 2009. Main results showed that: (1) Compared with AMB, hyacinth mulching significantly increased N concentration in rice plant over the season, hyacinth mulching significantly increased nitrogen accumulation in rice plant, and the increasing rate was gradual addition from mid-tillering to heading, but from heading to maturity the increasing rate was gradual decline. (2) Hyacinth mulching had no obvious effect on nitrogen allocation pattern in leaves and stems of rice over the season, significantly decreased nitrogen allocation pattern in spikes. (3) Except mid-tillering, hyacinth mulching resulted in the significant decrease in N use efficiency for biomass (NUEp) over the season and in N use efficiency for grain yield (NUEg) at grain maturity, but no effect was observed on nitrogen harvest index (NHI). (4) Nitrogen concentration and accumulation at most growth stages of rice increased with increasing N supply (p < 0.05 or 0.01), but NUEp and NUEg showed the opposite trends. 1(5) nteractions between hyacinth mulching x N were not observed for N uptake and utilization. Hyacinth mulching increased N concentration and N uptake, decreased NUEp and NUEg at most growth stages of rice.

[Treatment of lumbar spinal stenosis in the elder by micro-endoscopic discectomy].

OBJECTIVE: To explore the clinical effects of micro-endoscopic discectomy(MED) in treatment of lumbar spinal stenosis in the elder. METHODS: The data of 376 cases of the lumbar spinal stenosis were treated by MED from December 2003 to December 2009, 47 cases of them aged above 60 years old. Of them, 43 old patients were followed up for 3 months to 6 years. There were 27 males and 16 females,with an average age of 68.3 years old ranging from 60 to 91 years. The average age was 68.3 years. The course was from 3 months to 15 years (averaged 3.5 years). The clinical effects of postoperation were observed and evaluated according to Nakai standard. RESULTS: Forty-three patients were followed up for 3 months to 6 years. The incision of all the patients had no infection and achieved primary healing. The clinical effect of the patients were evaluated by Nakai standard. The results were excellent in 26 cases,good in 12,fair in 5. CONCLUSION: MED has characteristics of less injury,fast recovery and thorough decompression to nerve roots. It is a kind of good method for treating lumbar spinal stenosis in the elder if the indication were grasped strictly.

[Effects of free-air CO2 enrichment on nitrogen uptake and utilization of wheat].

In this paper, the effects of free-air CO2 enrichment (FACE) and its interaction with nitrogen supply on the nitrogen content, uptake, allocation, and use efficiency of winter wheat variety Ningmai 9 at its different growth stages were studied in 2001-2003. The results showed that under FACE treatment, the nitrogen content in wheat plant sampled at different growth stages all decreased significantly, with an average decrement of 4.4% to 13.4% compared with the control. The nitrogen accumulation under FACE increased significantly (9.2% -32.3%), and the increasing rate was larger at middle growth stage than at early and late growth stages. Nitrogen fraction was higher in stem, but lower in leaf. As for spike, the accumulation of nitrogen depended on the growth stage. FACE resulted in a significant increase (5.5% -10.3%) of nitrogen use efficiency for biomass production at all growth stages, of nitrogen harvest index (16.3%) at maturing stage, and of nitrogen use efficiency (9.3%) for grain yield. Nitrogen application increased the nitrogen content of wheat plant and its N uptake at all growth stages, decreased the nitrogen use efficiency, but had less effect on the nitrogen allocation in different organs.

(Piperazin-1-ium-κN)tris-(thio-cyanato-κN)zinc(II).

Hydro-thermal reaction of NaSCN, piperazine, Zn(II) and 2,6-naphthalenedicarboxylic acid in aqueous solutions gave rise to the title complex, [Zn(NCS)(3)(C(4)H(11)N(2))]. The Zn(II) atom is four-coordinate with distorted tetra-hedral geometry and lies in a mirror plane. N-H⋯S hydrogen bonds assemble the mol-ecules to form a three-dimensional framework.

[Landmark of facial nerve in middle ear surgery].

OBJECTIVE: To re-evaluate landmarks for facial nerve in middle ear surgery through temporal bone dissection and facial nerve surgery. METHODS: Some relative landmarks were found through 44 facial nerves dissection in cadaver and 106 cases of facial nerve decompression surgery. RESULTS: (1) Landmarks for vertical segment of the facial nerve: the vertical line in combined point between posterior and middle 1/3 horizontal semicircular canal clews the posterior edge of facial nerve; the prolong line of superior radian of incus short process clues to the anterior edge of the facial nerve, the facial nerve and horizontal semicircular canal are almost in the same plane. (2) Landmarks for horizontal segment of the facial nerve: the facial nerve tracks forward inferior to short process of incus and anterior to horizontal semicircular canal carina in 30 angel. The facial nerve, locating posterior and superior to cochleariform process and parallel with it, forms the step of middle-superior tympanic cavity and tracks forward to geniculate ganglion. (3) location of geniculate ganglion: The same distance prolong line of stapes head to cochleariform process clues to geniculate ganglion. (4) Location of the chorda tympani nerve: chorda tympani nerve, leaving tympanic sulcus at 3 clock of bone canaline left ear and at 9 clock of bone canaline right ear, tracks forward along tympani sulcus and then cross between long process of incus and manubrium. It lies in the border of pars tensa and pars flaccid and is about 5 - 8 mm from the stylomastoid foramen to where the chorda tympani nerve leaves the facial nerve. There is no difference of facial nerve structure in temporal bone dissection and in surgery. CONCLUSIONS: The fixed landmarks of middle ear are the frame of reference of facial nerve, in which horizontal semicircular canal is most invariable; and the safety of surgery will be improved by the reference of the facial nerve.