[Transcriptome analysis of seed embryo in dormancy and dormancy release state of Thesium chinense].

We used exogenous GA_3 to break the seed dormancy of Thesium chinense. We used high-throughput sequencing technology was used to sequence the transcriptome of dormant seed embryos and dormancy breaking seed embryos of Th. chinense, and the data was analyzed bioinformatically and systematically. The results showed that exogenous GA_3 could effectively break the seed dormancy of Th. chinense; 73 794 up-regulated genes and 42 776 down regulated genes were obtained by transcriptome sequencing; 116 570 diffe-rential genes were annotated by GO function to GO items such as metabolism process, cell process, cell, cell component, binding and catalytic activity. A total of 133 metabolic pathways were found by Pathway analysis of 26 508 differentially expressed genes. In the process of dormancy release, DEGs were mainly enriched in translation, carbohydrate metabolism, folding, classification, degradation and amino acid metabolism. Based on the annotation results in KEGG database, 20 metabolic pathways related to dormancy release were found. Dormancy release of Th. chinense seeds is a complex biological process, including cell morphology construction, secondary metabolite synthesis, sugar metabolism and plant signal transduction, among which plant hormone signal transduction is one of the key factors to regulate dormancy release. The results of qRT-PCR showed that the sequencing results were consistent with the actual results.

[Comparative analysis of active components and transcriptome between autotetraploid and diploid of Dendrobium huoshanense].

In this study, the roots, stems and leaves of diploid and autotetraploid Dendrobium huoshanense were used as materials to compare their contents of polysaccharides and alkaloids, and the transcriptome sequencing analysis was carried out. The results showed that the contents of polysaccharides and alkaloids in the roots, stems and leaves of tetraploid were 7.6%, 34.5%, 17.2%, 0.01%, 0.024% and 0.035% higher than those of diploid D. huoshanense, respectively. The contents of active components in different tissues were significantly different. There were 3 687 differentially expressed genes in diploid and tetraploid D. huoshanense, of which 2 346 genes were up-regulated and 1 341 down regulated. Go functional analysis showed that these genes were mainly involved in growth and development, stress resistance and other related functions. KEGG pathway analysis showed that most of the differential genes were concentrated in the processes of carbon metabolism, signal transduction, carbohydrate metabolism, amino acid metabolism and energy metabolism. The differential expression of key genes involved in the metabolism of polysaccharides, terpenes and polyketones, amino acid metabolism, hormone synthesis and signal transduction in diploid and tetraploid plants may be the main reason for the high energy content, the increase of active components and the growth potential of tetraploid plants.

[Identification and differential expression of miRNA related to seed dormancy of Paris polyphylla var. chinensis].

The purpose of this study was to explore the expression pattern of miRNA in the process of embryo dormancy and provide a reference for the mechanism of regulating seed dormancy and germination by miRNA. We used high-throughput sequencing technology, bioinformatics analysis and real-time fluorescent quantitative PCR(qPCR) technology to sequence, screen and identify miRNAs of dormant and dormant embryos. The results showed that there were 23 811 977, 24 276 695, 20 611 876 and 20 601 811 unique sequences in the four sample libraries during the period of dormancy and dormancy release. MiRNAs are mainly distributed between 21 and 24 nt, among which the length of 24 nt occurred most frequently. A total of 31 known miRNAs were identified, belonging to 13 different families. 93 new miRNAs were predicted by bioinformatics software. Ten miRNAs(mir156 a-5 p, mir160 a-5 p, mir160 h-1, mir169 a-5 p, mir157 d, mir159 a-1, mir395-3, mir156 f-5 p, mir156-2 and mir171 a-3 p) were screened out. In this study, 10 miRNAs related to seed dormancy release were identified. The target genes mainly involved carbohydrate metabolism, plant hormone signal transduction, cell division and growth. The results of qRT-PCR showed that the sequencing results were consistent with the actual results.

Unravelling the influence of sulfate loading on enhancing anaerobic co-digestion of corn stover and bio-kerosene production wastewater.

This study was conducted to investigate the effect of sulfate loading on methane production and organic matter degradation during the mesophilic anaerobic co-digestion of corn stover and bio-kerosene production wastewater (BKPW). The highest methane production of 192.04 mL/gVS was obtained at a sulfate concentration of 86 mg/L. This was 46.80% higher than that achieved by a sulfate concentration of 113 mg/L. Additional degradation of organic matter was obtained at a sulfate concentration of 113 mg/L because organic matter in the corn stover and BKPW was oxidized by sulfate-reducing bacteria (SRB). The concentration of sulfate declined by approximately 23% after 29 days of anaerobic co-digestion, and this reduction in sulfate was enhanced when the soluble chemical oxygen demand (SCOD)/sulfate ratio was less than 15. The results of a mass balance analysis showed that 34.87% of C element and 10.04% of S element in substrate, respectively, were converted to biogas during anaerobic co-digestion of corn stover and BKPW at a sulfate concentration of 86 mg/L. The microbial community was analysed using 16S rDNA sequencing technology, and the results showed that the relative abundance of Synergistes (related to methane production with acetic acid) at a sulfate concentration of 86 mg/L had obviously increased and was approximately 287% higher than the abundance achieved at a sulfate concentration of 113 mg/L.

Yangzheng Sanjie decoction regulates proliferation and apoptosis of gastric cancer cells by enhancing let-7a expression.

AIM: To explore the let-7a-mediated anti-cancer effect of Yangzheng Sanjie decoction (YZSJD) in gastric cancer (GC) cells. METHODS: YZSJD-containing serum (YCS) was prepared using traditional Chinese medicine serum pharmacology methods. After YCS treatment, cell proliferation and apoptosis were assessed by cell counting kit-8 assay and flow cytometry, respectively, and miRNA expression profiles were determined using qPCR arrays. Let-7a expression was examined by in situ hybridization in GC tissues and by qPCR in GC cells. c-Myc protein expression was detected by immunohistochemistry in GC tissues, and by Western blot in cell lines. RESULTS: YZSJD significantly inhibited proliferation and induced apoptosis in AGS and HS-746T GC cells. After treatment with YCS, the miRNA expression profiles were altered and the reduced let-7a levels in both cell lines were up-regulated, accompanied by a decrease in c-Myc expression. Moreover, decreased let-7a expression and increased c-Myc expression were observed during the progression of gastric mucosa cancerization. CONCLUSION: YZSJD inhibits proliferation and induces apoptosis of GC cells by restoring the aberrant expression of let-7a and c-Myc.

[Transport Loss Estimation of Fine Particulate Matter in Sampling Tube Based on Numerical Computation].

In-situ measurement of PM2.5 physical and chemical properties is a substantial approach for the mechanism investigation of PM2.5 pollution. Minimizing PM2.5 transport loss in sampling tube is essential for ensuring the accuracy of the measurement result. In order to estimate the integrated PM2.5 transport efficiency in sampling tube and optimize tube designs, the effects of different tube factors (length, bore size and bend number) on the PM2.5 transport were analyzed based on numerical computation. The results showed that PM2.5 mass concentration transport efficiency of vertical tube with flow rate at 20.0 L·min-1, bore size at 4 mm, length at 1.0 m was 89.6%. However, the transport efficiency increased to 98.3% when the bore size increased to 14 mm. PM2.5 mass concentration transport efficiency of horizontal tube with flow rate at 1.0 L·min-1, bore size at 4 mm, length at 10.0 m was 86.7%, and increased to 99.2% with length at 0.5 m. Low transport efficiency of 85.2% for PM2.5 mass concentration was estimated in bend with flow rate at 20.0 L·min-1, bore size at 4 mm, curvature angle at 90°. Laminar flow of air in tube through keeping the ratio of flow rate (L·min-1) and bore size (mm) below 1.4 was beneficial to decrease the PM2.5 transport loss. For the target of PM2.5 transport efficiency higher than 97%, it was advised to use vertical sampling tubes with length less than 6.0 m for the flow rates of 2.5, 5.0, 10.0 L·min-1 and bore size larger than 12 mm for the flow rates of 16.7 or 20.0 L·min-1. For horizontal sampling tubes, tube length was decided by the ratio of flow rate and bore size. Meanwhile, it was suggested to decrease the amount of the bends in tube of turbulent flow.

[The protective effect of PEP-1-SOD1 preconditioning on hypoxia/reoxygenation injury in cultured human umbilical vein endothelial cells].

OBJECTIVE: To construct prokaryotic expression vector of pET15b-PEP-1-SOD1 and investigate whether PEP-1-SOD1 fusion protein could be transduced into human umbilical vein endothelial cells (HUVECs) and the effects on hypoxia/reoxygenation injury. METHODS: The recombinant plasmids pET15b-SOD1 and pET15b-PEP-1-SOD1 were constructed and transformed into E. coli BL21 (DE3) to express SOD1 and PEP-1-SOD1 with an N-terminal His-tag. The purified SOD1 and PEP-1-SOD1 were incubated with HUVECs and the viability (MTT assay) and the release of lactate dehydrogenase (LDH) in culture medium were determined in the hypoxia/reoxygenation injury model. The morphological changes were observed under an inverted phase contrast microscope. The content of malondialdehyde (MDA) in HUVECs was also determined with the method of thiobarbituric acid. RESULTS: PEP-1-SOD1 fusion protein could be transduced into cultured HUVECs in a time- and dose-dependent manner. The intracellular enzymatic activity of PEP-1-SOD1 after 30 min incubation with HUVECs was significantly higher than control group (60.88 U/ml +/- 6.73 U/ml vs. 41.06 U/ml +/- 4.19 U/ml, P < 0.01). The transduced PEP-1-SOD1 protein was enzymatically stable for 24 h within cells. After hypoxia/reoxygenation injury, control HUVECs shrunk, became round-shaped and intercellular space increased, while these morphological changes were not observed in PEP-1-SOD1 transduced HUVECs. PEP-1-SOD1 transduction also markedly increased the viability, decreased LDH leakage into culture media and reduced the content of MDA post hypoxia/reoxygenation. CONCLUSIONS: PEP-1-SOD1 fusion protein could be efficiently transduced into HUVECs in a natively active form, and the delivered enzymatically active PEP-1-SOD1 exhibits cellular protection against hypoxia/reoxygenation injury in HUVECs. The transduction of SOD1 mediated by cell-penetrating peptide, PEP-1, provides a basis for further research on the prevention of ischemia/reperfusion injury in vivo.