Full-length transcriptome characterization of Platycladus orientalis based on the PacBio platform.

As a unique and native conifer in China, Platycladus orientalis is widely used in soil erosion control, garden landscapes, timber, and traditional Chinese medicine. However, due to the lack of reference genome and transcriptome, it is limited to the further molecular mechanism research and gene function mining. To develop a full-length reference transcriptome, tissues from five different parts of P. orientalis and four cone developmental stages were sequenced and analyzed by single-molecule real-time (SMRT) sequencing through the PacBio platform in this study. Overall, 37,111 isoforms were detected by PacBio with an N50 length of 2,317 nt, an average length of 1,999 bp, and the GC content of 41.81%. Meanwhile, 36,120 coding sequences, 5,645 simple sequence repeats (SSRs), 1,201 non-coding RNAs (lncRNAs), and 182 alternative splicing (AS) events with five types were identified using the results obtained from the PacBio transcript isoforms. Furthermore, 1,659 transcription factors (TFs) were detected and belonged to 51 TF families. A total of 35,689 transcripts (96.17%) were annotated through the NCBI nr, KOG, Swiss-Prot and KEGG databases, and 385 transcript isoforms related to 8 types of hormones were identified incorporated into plant hormone signal transduction pathways. The assembly and revelation of the full-length transcriptome of P. orientalis offer a pioneering insight for future investigations into gene function and genetic breeding within Platycladus species.

[Influence of long non-coding RNA Gm15645 on the podocyte injury in diabetic nephropathy].

OBJECTIVE: To explore the influence of long non-coding (lnc) RNA Gm15645 on the podocyte injury in mice with diabetic nephropathy. METHODS: Male db/db mice (with Type 2 diabetes) with a genetic background of C57BLKs/J and db/m mice (healthy) born in littermates were randomly divided into three groups. db/db group was injected with lncRNAGm15645 shRNA lentivirus with a podocyte-specific marker NPHS2; db/db blank group was injected with saline, and db/db control group was injected withnon-sense lentivirus. The results of PAS staining, pathological changes of renal tissue, relative expression of GSK-3beta, and podocin expression were compared. RESULTS: lncRNAGm15 645 was overexpressed and podocin was down-regulated in the lentivirus overexpressed group. Mesangial cell proliferation, mesangial matrix hyperplasia, thickened basement membrane, widely fused foot process, and podocyte injury were observed by PAS staining. The expression of Gm15645 in the db/db group was significantly lower than that of the db/db blank group and db/db control group (P< 0.05), while the expression of podocin was higher (P< 0.05). Gm15645 was co-stained with podocin in renal tissue, and the target gene was GSK-3beta. CONCLUSION: lncRNAGm15645 may provide an early biomarker for the occurrence of podocyte injury in diabetic nephropathy. The mechanism may be related to the feedback regulation of GSK-3beta gene.

MiR-429 prohibited NF-κB signalling to alleviate contrast-induced acute kidney injury via targeting PDCD4.

MiR-429 was reported to be downregulated in contrast-induced acute kidney injury (CI-AKI). However, whether miR-429 is functionally relevant with CI-AKI needs further investigation. Human renal tubular epithelial cell (HK-2) cells were stimulated with contrast media iodixanol to establish in vitro CI-AKI model. Cell Counting Kit-8 (CCK-8) was applied to access cell viability. Flow cytometry was performed to determine apoptosis. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to evaluate level of programmed cell death 4 (PDCD4) mRNA and miR-429 while western blot was applied to evaluate level of proteins including PDCD4, B-cell leukaemia/lymphoma 2 (Bcl-2), BCL2-associated X protein (Bax), cleaved caspase 3, cleaved caspase 9, p65, phosphorylated p65. Dual luciferase assay was used to validate miR-429 targeting PDCD4. MiR-429 was downregulated whereas PDCD4 was upregulated in contrast media iodixanol-stimulated HK-2 cells. MiR-429 overexpression elevated cell viability and attenuated cell apoptosis. Moreover, the activation of nuclear factor kappa-B (NF-κB) signalling was suppressed after miR-429 overexpression, while PDCD4 overexpression reversed these effects. MiR-429 directly targeted PDCD4 and negatively regulated its expression. CI-AKI induced NF-κB signalling activation and PDCD4 overexpression further promoted NF-κB signalling activation. However, the treatment of BAY11-7082 reversed above results. Overexpression of miR-429 attenuated apoptosis and elevated cell viability in a CI-AKI cell model via targeting PDCD4 and thus restraining NF-κB signalling.

Long-term efficacy of different procedures for treatment of varicose veins: A network meta-analysis.

BACKGROUND: Various procedures for the treatment of varicose veins have been shown to have long-term effectiveness, but research has yet to identify the most effective procedure. The aim of this study was to investigate the long-term efficacy of different procedures based on Bayesian network meta-analysis and to rank therapeutic options for clinical decision-making. METHODS: Globally recognized databases, namely, MEDLINE, Embase, and Cochrane Central, were searched for randomized controlled trials (RCTs). Quantitative pooled estimation of successful treatment rate (STR) and recurrence rate (RR) was performed to assess the long-term efficacy of each procedure with more than a 1-year follow-up. The surface under the cumulative ranking (SUCRA) probabilities of the P values regarding STR and RR were calculated to rank various procedures. Grades of Recommendations Assessment, Development and Evaluation (GRADE) criteria were utilized for the recommendation of evidence from pairwise direct comparisons. RESULTS: A total of 39 RCTs encompassing a total of 6917 limbs were eligible and provided relative raw data. After quantitative analysis, the CHIVA procedure was determined to have the best long-term efficacy, as it had the highest STR (SUCRA, 0.37). Additionally, the results revealed that CHIVA possessed the highest probability of achieving the lowest long-term RR (SUCRA, 0.61). Moreover, the sensitivity analysis with inconsistency approach clarified the reliability of the main results, and the evidence of most direct comparisons was ranked as high or moderate. CONCLUSION: CHIVA seemed to have superior clinical benefits on long-term efficacy for treating varicose veins. However, the conclusion still needs additional trials for supporting evidence.

Daphnetin inhibits high glucose-induced extracellular matrix accumulation, oxidative stress and inflammation in human glomerular mesangial cells.

Diabetic nephropathy (DN) is one of the most common causes of end-stage renal disease (ESRD). Oxidative stress and inflammation have been documented to play important roles in the pathogenesis of DN. Daphnetin, a natural coumarin compound, possesses antioxidant and anti-inflammatory activities. However, the role of daphnetin in DN has not yet been investigated. The aim of the present study was to explore the function of daphnetin in DN and the underlying mechanism in vitro. Our results demonstrated that daphnetin alleviated cell proliferation induced by high glucose (HG) in human mesangial cells (MCs). Daphnetin strikingly reduced reactive oxygen species (ROS) and malonaldehyde (MDA) levels, and induced the superoxide dismutase (SOD) activity in HG-stimulated MCs. Besides, the production of TNF-α, IL-1ß, IL-6, fibronectin (FN) and collagen IV (Col IV) was also inhibited by daphnetin in HG-stimulated MCs. In addition, daphnetin enhanced the expression of nuclear factor-erythroid 2-related factor 2 (Nrf2) and inhibited the levels of p-Akt and p-p65 in HG-stimulated MCs. The results indicated that daphnetin inhibited HG-induced oxidative stress, inflammatory response, and ECM accumulation in human MCs. The effect is partially mediated by Nrf2/keap1 and Akt/NF-κB pathways. The findings suggested that daphnetin might be a therapeutic or preventive agent for DN.

An improved method for high-temperature induced embryo sac chromosome doubling in Populus simonii Carr. × P. nigra var. italica (Moench.) Kochne, an interspecific hybrid of Tacamahaca and Aigeiros poplars.

Chromosome doubling is considered an important technique in poplar breeding, with many triploid clones being artificially induced and selected for promotion in the north and northeast of China because of their outstanding traits in vegetative growth and environmental adaption. In this study, the triploid yield of Populus simonii Carr × P. nigra var. italica (Moench.) Kochne was 23.41%, which exceeded the yield attained in our previous studies due to the use of an optimized method of chromosome doubling in the embryo sac at a high temperature. The development of the embryo sac after the pollination of this hybrid was investigated to determine the induction period. Ploidy of seedlings was identified by flow cytometry after initial filtering using the chloroplast counting method. Eleven triploids and one tetraploid were ultimately obtained, and the optimal operating conditions were exposure of female catkins to 41 °C for 2 h at 66 h after pollination (HAP). This study identified an efficient method of chromosome doubling in P. simonii × P. nigra var. italica and provided several polyploids for Populus polyploid breeding programs and subsequent studies.

Inducing triploids and tetraploids with high temperatures in Populus sect. Tacamahaca.

KEY MESSAGE: This study is the first to report that triploids and tetraploids have been successfully produced through embryo sac and zygotic embryo chromosome doubling with high temperatures in P. simonii Carr. and its hybrid. A new synthetic polyploid induced by hybridization with unreduced gametes and heterozygotic embryo chromosome doubling can effectively combine polyploidy and heterosis, which can provide two major breeding advantages. In Populus, successfully creating and cultivating new polyploid varieties have economic and ecological production value. This was the first successful study in which embryo sac and zygotic embryo chromosome doubling was induced using high temperatures to produce triploids and tetraploids in Populus simonii Carr. and its hybrid, P. simonii × P. nigra var. Italica, of Populus sect. Tacamahaca. The relationship between flower bud morphological characteristics (time after pollination) and female meiotic stage (embryo sac and zygotic embryo development) was established to guide the induction treatment period. In the resulting progeny, 37 triploids and 12 tetraploids were obtained and identified using flow cytometry. The optimal temperatures for embryo sac and zygotic embryo chromosome doubling were 38 and 41 °C, respectively. Cytogenetic analysis revealed that 66-72 h after pollination (HAP), a period characterized by a high proportion of one-nucleate and two-nucleate embryo sacs, was the optimal period for embryo sac chromosome doubling. For zygotic embryo chromosome doubling, 168 HAP was the optimal induction period, as there was a high proportion of two-cell and four-cell proembryos. The results indicate that inducing embryo sac and zygotic embryo chromosome doubling is an ideal method for producing polyploids. The methods for inducing polyploids and for evaluating ploidy and offspring with different ploidies and heterozygosity in this study will be useful for genetic research and Populus breeding programmes.

Thiazolidinedione attenuate proteinuria and glomerulosclerosis in Adriamycin-induced nephropathy rats via slit diaphragm protection.

AIM: The slit diaphragm (SD) of podocyte impairment contributes to massive proteinuria and progressive glomerulosclerosis in many human glomerular diseases. The aim of the study was to determine if thiazolidinedione (TZD) reduce proteinuria and glomerulosclerosis in focal segmental glomerulosclerosis (FSGS) by preserving the structure and function of SD. METHODS: Adriamycin-induced FSGS rat models were employed. Urinary protein content was measured dynamically during the experiment. Additional biochemical parameters in serum samples were measured after the animals were killed. Glomerular sclerosis index (SI) and podocyte foot processes fusion rate (PFR) were evaluated. The protein and mRNA expressing levels of nephrin, podocin and CD2-associated protein (CD2AP) in glomeruli were assessed by immunohistochemistry and real-time quantitative polymerase chain reaction, respectively. The density of podocytes was also evaluated after anti-Wilms' tumour-1 immunohistochemical staining. RESULTS: Rosiglitazone treatment partially reduced proteinuria, but did not significantly affect the serum levels of triglyceride, cholesterol, albumin, glucose, urea nitrogen and creatinine in Adriamycin-induced FSGS rats. Glomerular SI and podocyte foot PFR were significantly attenuated by rosiglitazone treatment. Rosiglitazone prevented the reduction of nephrin, podocin and CD2AP protein expression induced by Adriamycin, however, the mRNA expression levels of these SD-related markers did not change significantly. Rosiglitazone therapy did not reverse Adriamycin-mediated reduction of the density of podocytes. CONCLUSIONS: The study data suggest that TZD are promising therapeutic agents on FSGS, and the mechanism may be mediated in part by directly protecting the structure and function of SD.

Strategies comparison of eliminating the passivation of non-aromatic intermediates in degradation of Orange II by Fe3+/H2O2.

The degradation kinetics of Orange II by Fe(3+)/H(2)O(2) was examined under various conditions in the presence of the non-aromatic organic intermediates such as malonic acid (MA), oxalic acid (OX) and EDTA. Various strategies were adopted to eliminate the passivation of the non-aromatic organic intermediates in the Fenton-like degradation of Orange II. UV irradiation, visible irradiation, hydroquinone (HQ) additive and their hybrid were used to accelerate the decoloration of Orange II; however, some of them were failed to show a good performance in the presence of non-aromatic organics, especially for the mineralization of Orange II. Aluminum ions additive released the ferric ions via a competitive chelation process, and accelerated the reaction obviously. From the decoloration and the mineralization data of Orange II, UV irradiation combined with Al(3+) ion additive was regarded as the most favorable strategy for deep mineralization of Orange II by Fe(3+)/H(2)O(2) in the presence of non-aromatic intermediates. However, by considering the decoloration of Orange II with moderate mineralization, the combination of visible irradiation and Al(3+) ion additive might present the best performance.

5-{2-[(2-Hydroxy-5-methyl-phen-yl)(phenyl)methyl-eneamino]phenyl-imino-meth-yl}-pyrrole-2-carbaldehyde.

The title compound, C(26)H(21)N(3)O(2), is an unsymmetrical tetra-dentate Schiff base ligand. The hydr-oxy group forms an intra-molecular O-H⋯N hydrogen bond with an adjacent N atom. An inter-molecular N-H⋯O hydrogen bond creates centrosymmetric dimers in the crystal packing.

N-(4-Cyano-benz-yl)benzamide.

The title compound, C(15)H(12)N(2)O, is a derivative of 4-(amino-meth-yl)benzonitrile, an important pestcide inter-mediate. In the crystal structure, mol-ecules are linked via inter-molecular N-H⋯O hydrogen bonds, forming infinite chains.