Upregulation of miR-199a-5p Protects Spinal Cord Against Ischemia/Reperfusion-Induced Injury via Downregulation of ECE1 in Rat

Ischemia–reperfusion (I/R)-induced spinal cord injury can cause apoptotic damage and subsequently act as a blood–spinal cord barrier damage. MicroRNAs (miRNAs) contributed to the process of I/R injury by regulating their target mRNAs. miR-199a-5p is involved in brain and heart I/R injury; however, its function in the spinal cord is not yet completely clarified. In this study, we investigated the role of miR-199a-5p on spinal cord I/R via the endothelin-converting enzyme 1, especially the apoptosis pathway. In the current study, the rat spinal cord I/R injury model was established, and the Basso Beattie Bresnahan scoring, Evans blue staining, HE staining, and TUNEL assay were used to assess the I/R-induced spinal cord injury. The differentially expressed miRNAs were screened using microarray. miR-199a-5p was selected by unsupervised hierarchical clustering analysis. The dual-luciferase reporter assay was used for detecting the regulatory effects of miR-199a-5p on ECE1. In addition, neuron expression was detected by immunostaining assay, while the expressions of p-ERK, ERK, p-JNK, JNK, caspase-9, Bcl-2, and ECE1 were evaluated by Western blot. The results indicated the successful establishment of the I/R-induced spinal cord injury model; the I/R induced the damage to the lower limb motor. Furthermore, 18 differentially expressed miRNAs were detected in the I/R group compared to the sham group, and miR-199a-5p protected the rat spinal cord injury after I/R. Moreover, miR-199a-5p negatively regulated ECE1, and silencing the ECE1 gene also protected the rat spinal cord injury after I/R. miR-199a-5p or silencing of ECE1 also regulated the expressions of caspase-9, Bcl-2, p-JNK, p-ERK, and ECE1 in rat spinal cord injury after I/R. Therefore, we demonstrated that miR-199a-5p might protect the spinal cord against I/R-induced injury by negatively regulating the ECE1, which could aid in developing new therapeutic strategies for I/R-induced spinal cord injury.     

金丝李果实、种子形态分化及外源物质对种子萌发和幼苗生长的影响

通过采集3个天然种群的金丝李(Garcinia paucinervis)果实,观察果实和种子的形态性状,分析其种群内和种群间的形态分化,观察不同种群种子萌发和幼苗生长的规律;采用四种植物生长调节剂和两种化学药剂,研究浸种对金丝李种子萌发和幼苗生长的影响。结果表明:金丝李种子长2.48~3.08 cm、宽1.49~1.67cm、百粒重392.50~438.18 g;3个种群果实和种子已产生明显的形态分化,胡润种群多数表型性状值在3个种群中最小,安宁种群的果实和种子偏圆,弄岗种群偏窄;在种群内部,果实和种子重量的分化较显著,整体形态较稳定;在萌发过程中,种子留存,不定根逐渐取代胚根成为主根;3个种群的萌发过程均缓慢,中间阶段几乎停滞,出苗极不整齐,萌发率表现为安宁种群(78.33%)弄岗种群(55%)胡润种群(48.33%),种子存在休眠;从种子萌发来看,以80 mg·L~(-1)的6-BA浸种24 h处理效果最好,萌发时滞最短、萌发率最高、平均萌发时间最短;结合幼苗生长情况来看,10~50 mg·L~(-1)的6-BA和500 mg·L~(-1)的GA_3处理均适宜,以50 mg·L~(-1)的6-BA效果最佳。低浓度的KNO_3和NH_4NO_3能提高萌发率,但未显著加快萌发进程,IAA和NAA对种子萌发和幼苗生长有抑制作用。该研究结果为金丝李的遗传多样性提供了形态学资料,为更好地保护、开发和利用这一珍贵树种提供了科学依据。     

Mouse DNA polymerase kappa has a functional role in the repair of DNA strand breaks

The Y-family of DNA polymerases support of translesion DNA synthesis (TLS) associated with stalled DNA replication by DNA damage. Recently, a number of studies suggest that some specialized TLS polymerases also support other aspects of DNA metabolism beyond TLS in vivo. Here we show that mouse polymerase kappa (Polκ) could accumulate at laser-induced sites of damage in vivo resembling polymerases eta and iota. The recruitment was mediated through Polκ C-terminus which contains the PCNA-interacting peptide, ubiquitin zinc finger motif 2 and nuclear localization signal. Interestingly, this recruitment was significantly reduced in MSH2-deficient LoVo cells and Rad18-depleted cells. We further observed that Polκ-deficient mouse embryo fibroblasts were abnormally sensitive to H₂O₂ treatment and displayed defects in both single-strand break repair and double-strand break repair. We speculate that Polκ may have an important role in strand break repair following oxidative stress in vivo.     

Design,synthesis and in vitro anti-mycobacterial evaluation of gatifloxacin-1H-1,2,3-triazole-isatin hybrids

A set of novel gatifloxacin-1H-1,2,3-triazole-isatin hybrids 6a-l was designed, synthesized and evaluated for their in vitro anti-mycobacterial activities against M. tuberculosis (MTB) H₃₇Rv and MDR-TB as well as cytotoxicity. The results showed that all the targets (MIC: 0.025–3.12 μg/mL) exhibited excellent inhibitory activity against MTB H₃₇Rv and MDR-TB, but were much more toxic (CC₅₀: 7.8–62.5 μg/mL) than the parent gatifloxacin (GTFX) (CC₅₀: 125 μg/mL). Among them, 61 (MIC: 0.025 μg/mL) was 2–32 times more potent in vitro than the references INH (MIC: 0.05 μg/mL), GTFX (MIC: 0.78 μg/mL) and RIF (MIC: 0.39 μg/mL) against MTB H₃₇Rv. The most active conjugate 6 k (MIC: 0.06 μg/mL) was 16–>2048 times more potent than the three references (MIC: 1.0–>128 μg/mL) against MDR-TB. Both of the two hybrids warrant further investigations.     

Effects of nitrogen deposition and management practices on leaf litterfall and N and P return in a Moso bamboo forest

The effects of nitrogen (N) deposition and management practices on leaf litterfall and N and P return in Moso bamboo forest are not yet known. In this study, we investigated the effects of four levels of simulated N deposition, including low-N (N30, 30 kg ha^?1 year^?1), medium-N (N60, 60 kg ha^?1 year^?1), and high-N (N90, 90 kg ha^?1 year^?1), and a control with no N (N-free addition). The experiment was performed in a Moso bamboo forest under conventional management (CM) and intensive management (IM). The results showed that leaf litterfall and N and P return occurred mainly from March to June and accounted for 78.2–82.2, 78.5–82.1, and 85.6–94.6% of annual leaf litterfall, N return, and P return, respectively. Unlike CM, IM significantly increased leaf litterfall and N and P return. The positive effects were further amplified by low- and medium-N deposition, but not high-N deposition. The combination of low- and medium-N deposition and IM significantly increased N and P return, but not litterfall. Our results indicated that the interaction of anthropogenic management practices and N deposition need to be considered when estimating the effects of N deposition on the biogeochemical cycle of a forest ecosystem.     

The complete chloroplast genomes of three Hamamelidaceae species: Comparative and phylogenetic analyses

Hamamelidaceae is an important group that represents the origin and early evolution of angiosperms. Its plants have many uses, such as timber, medical, spice, and ornamental uses. In this study, the complete chloroplast genomes of Loropetalum chinense (R. Br.) Oliver, Corylopsis glandulifera Hemsl., and Corylopsis velutina Hand.‐Mazz. were sequenced using the Illumina NovaSeq 6000 platform. The sizes of the three chloroplast genomes were 159,402 bp (C. glandulifera), 159,414 bp (C. velutina), and 159,444 bp (L. chinense), respectively. These chloroplast genomes contained typical quadripartite structures with a pair of inverted repeat (IR) regions (26,283, 26,283, and 26,257 bp), a large single‐copy (LSC) region (88,134, 88,146, and 88,160 bp), and a small single‐copy (SSC) region (18,702, 18,702, and 18,770 bp). The chloroplast genomes encoded 132–133 genes, including 85–87 protein‐coding genes, 37–38 tRNA genes, and 8 rRNA genes. The coding regions were composed of 26,797, 26,574, and 26,415 codons, respectively, most of which ended in A/U. A total of 37–43 long repeats and 175–178 simple sequence repeats (SSRs) were identified, and the SSRs contained a higher number of A + T than G + C bases. The genome comparison showed that the IR regions were more conserved than the LSC or SSC regions, while the noncoding regions contained higher variability than the gene coding regions. Phylogenetic analyses revealed that species in the same genus tended to cluster together. Chunia Hung T. Chang, Mytilaria Lecomte, and Disanthus Maxim. may have diverged early and Corylopsis Siebold & Zucc. was closely related to Loropetalum R. Br. This study provides valuable information for further species identification, evolution, and phylogenetic studies of Hamamelidaceae plants.