[Effects of Long-term Fertilization on Soil Microbial Diversity and Community Structure in the Agro-pastoral Ecotone].

To explore the effects of different long-term fertilization treatments on soil microbial diversity and community structure in the drylands of an agro-pastoral ecotone, a long-term fertilization experiment at the Inner Mongolia cultivated land conservation science observation and experiment station, Ministry of Agriculture, and rural areas was taken as the research object. Four treatments, including no fertilizer (CK), single nitrogen fertilizer (NF), single chemical fertilizer (CF), and the combined application of organic manure and chemical fertilizer (CFM), were selected for the collection of 0-10 cm and 10-20 cm soil at potato maturity 16 years after the experiment (2019). High-throughput sequencing technology was used to assess the soil bacterial and fungal communities to explore the effects of different fertilization measures on soil quality from the perspective of microorganisms, and the partial least squares path model (PLS-PM) was used to reveal the key environmental driving factors of soil microbial community alternation and crop yield improvement in dryland during fertilization mode transformation. The results showed that:① the CF and CFM treatments significantly improved soil fertility, but the effect of the latter was significantly better than that of the former. Soil available nitrogen, available phosphorus, and available potassium in the CFM treatment increased by 131.9%-174.7%, 216.9%-283.3%, and 103.3%-109.3%, respectively, and organic matter and total nitrogen content also increased significantly. The CF treatment still maintained a high soil pH, whereas the NF treatment significantly decreased soil pH and had little effect in improving soil fertility. ② Compared with that under CK, the NF treatment significantly reduced the soil bacterial Chao1 and Shannon index, and the CFM treatment significantly increased the soil bacterial species richness, Chao1 index, and soil fungal Shannon index, whereas soil bacterial and fungal diversity in the CF treatment did not reach a significant difference level with CK. ③ The soil microbial community composition at 0-10 cm and 10-20 cm was similar. The CFM treatment increased the relative abundance of soil beneficial bacteria and decreased the relative abundance of pathogenic bacteria. The relative abundance of dominant bacteria such as Proteobacteria, Bacteroidetes, and Gemmatimonadetes increased. The relative abundances of Actinobacteria, Ascomycota, and Basidiomycota were decreased, whereas the NF and CF treatments showed the opposite trend. ④ PLS-PM analysis showed that with the gradual change in fertilization mode from CK→NF→CF→CFM, the driving factors affecting microbial community succession and yield increase were also changed from soil pH→soil NPK content→soil pH, SOM, and NPK content. In general, long-term fertilization had significant effects on soil chemical properties and microbial communities in drylands in the agro-pastoral ecotone. As the optimal fertilization choice, CFM was significantly better than NF and CF in improving soil fertility and inhibiting the growth of pathogenic microorganisms. The number of pathogens in long-term non-fertilization and unbalanced fertilization soil was significantly increased, and the risk of crop infection to indigenous diseases was increased. The research results can provide scientific reference for farmland nutrient balance management and soil microenvironment improvement of the agricultural ecosystem in the agro-pastoral ecotone in North China.

Influence of Shielding Gas on Microstructure and Properties of GMAW DSS2205 Welded Joints.

In the present study, the microstructures and properties of DSS 2205 solid wire MIG welded samples prepared in different shielding gases (pure Ar gas, 98%Ar + 2%O2 and 98%Ar + 2%N2) were investigated for improving the weldability of DSS 2205 welded joint. The work was conducted by mechanical property tests (hardness and tensile test) and corrosion resistance property tests (immersion and electrochemical tests). The results show that adding 2%O2 into pure Ar gas as the shielding gas decreases crystal defects (faults) and improves the mechanical properties and corrosion resistance of the welded joints. Phase equilibrium and microstructural homogeneity in welded seam (WS) and heat-affected zone (HAZ) can be adjusted and the strength and corrosion resistance of welded joints increased obviously by adding 2%N2 to pure Ar gas as the shielding gas. Compared with DSS 2205 solid wire MIG welding in 98%Ar + 2%O2 mixed atmosphere, the strength and corrosion resistance of welded joints are improved more obviously in 98%Ar + 2%N2 mixed atmosphere.

Impacts of Stress Relief Treatments on Microstructure, Mechanical and Corrosion Properties of Metal Active-Gas Welding Joint of 2205 Duplex Stainless Steel.

Stress relief treatments were carried out separately with a pneumatic chipping hammer, ultrasonic peening treatment, and heat treatment for metal active-gas welding (MAG) welded joints of 2205 duplex stainless steel. The effects of these methods on the residual stress, microstructure, mechanical properties and corrosion resistance of welded joints were studied. Results show the stress state of the weld and the surrounding area was effectively improved by the pneumatic chipping hammer and ultrasonic peening treatment, and the residual stress field of the surface layer changed from tensile stress to compressive stress. On the contrary, low-temperature stress relieving annealing had no obvious effect on stress distribution. After the pneumatic chipping hammer and ultrasonic peening treatment, the welded joints were machined and hardened. Correspondingly, strength and hardness were improved. However, the heat treatment only led to a slight decrease in strength and hardness due to the static recovery of the welded joint structure. All stress relief methods effectively improved the corrosion resistance of welded joints, with the ultrasonic peening treatment giving the best performance.

MicroRNA-581 promotes hepatitis B virus surface antigen expression by targeting Dicer and EDEM1.

Hepatitis B virus surface antigen (HBsAg) is an important risk factor for hepatocellular carcinoma (HCC) and is downregulated during hepatocarcinogenesis. MicroRNAs (miRNAs) are frequently deregulated in HCC tissues. However, whether the deregulation of certain miRNAs in HCC has an impact on HBsAg expression remains unclear. We found here that microRNA-581 (miR-581), which is deregulated during hepatocarcinogenesis, promoted HBsAg expression. Additionally, miR-581 targeted Dicer and endoplasmic reticulum degradation-enhancing alpha-mannosidase-like protein 1 (EDEM1) and repressed their expression. Although Dicer cannot process HBV transcripts, Dicer knockdown led to increased HBsAg secretion, most likely due to a reduction in the levels of Dicer-processed 7SL RNA fragments. Moreover, Dicer-processed 7SL RNA fragments partially inhibited the ability of miR-581 to stimulate HBsAg expression. Furthermore, we found that forced EDEM1 expression inhibited miR-581-mediated induction of HBsAg. Finally, transfection of miR-581 into HepG2.2.15 cells promoted cell proliferation and led to upregulation of genes involved in development, cell proliferation and protein secretion. Altogether, we conclude that miR-581 promotes HBsAg expression by targeting Dicer and EDEM1. Our findings suggest that downregulation of miR-581 during hepatocarcinogenesis may lead to a reduction in HBsAg expression and impede HCC development.

Hepatitis B surface antigen inhibits MICA and MICB expression via induction of cellular miRNAs in hepatocellular carcinoma cells.

Hepatitis B surface antigen (HBsAg) seropositivity is an important risk factor for hepatocellular carcinoma (HCC), and HBsAg-transgenic mice have been reported to spontaneously develop HCC. The major histocompatibility complex class I-related molecules A and B (MICA and MICB) are NKG2D ligands that play important roles in tumor immune surveillance. In the present study, we found that HBsAg overexpression in HepG2 cells led to upregulation of 133 and downregulation of 9 microRNAs (miRNAs). Interestingly, several HBsAg-induced miRNAs repressed the expression of MICA and MICB via targeting their 3'-untranslated regions. In addition, the expression of MICA and MICB was significantly reduced upon HBsAg overexpression, which was partially restored by inhibiting the activities of HBsAg-induced miRNAs. Moreover, HBsAg-overexpressing HCC cells exhibited reduced sensitivity to natural killer cell-mediated cytolysis. Taken together, our data suggest that HBsAg supresses the expression of MICA and MICB via induction of cellular miRNAs, thereby preventing NKG2D-mediated elimination of HCC cells.

Decreased dicer expression enhances SRP-mediated protein targeting.

We have shown that Dicer processes 7SL RNA into different fragments ranging from ∼20 to more than 200 nucleotides. Here we addressed the molecular functions of these 7SL RNA fragments and found that some of them functioned as dominant-negative regulators of the full-length 7SL RNA, interfering with signal recognition particle (SRP) complex formation. Transfection of these 7SL RNA fragments inhibited the expression of cell surface glycoproteins, the targeting of a reporter protein to the endoplasmic reticulum, and the secretion of secreted alkaline phosphatase. These results suggest that some Dicer-processed 7SL RNA fragments interfered with SRP-mediated protein targeting. Moreover, we showed that Dicer knockdown enhanced SRP-mediated protein targeting and that transfection of a mixture of the 7SL RNA fragments partially restored this effect. Our data indicate that Dicer can fine-tune the efficiency of SRP-mediated protein targeting via processing a proportion of 7SL RNA into fragments of different lengths.

Dicer-dependent biogenesis of small RNAs derived from 7SL RNA.

It has been reported that decreased Dicer expression leads to Alu RNAs accumulation in human retinal pigmented epithelium cells, and Dicer may process the endogenous SINE/B1 RNAs (the rodent equivalent of the primate Alu RNAs) into small interfering RNAs (siRNAs). In this study, we aimed to address whether Dicer can process Alu RNAs and their common ancestor, 7SL RNA. Using Solexa sequencing technology, we showed that Alu-derived small RNAs accounted for 0.6% of the total cellular small RNAs in HepG2.2.15 cells, and the abundance decreased when Dicer was knocked down. However, Alu-derived small RNAs showed different characteristics from miRNAs and siRNAs, the classic Dicer-processed products. Interestingly, we found that small RNAs derived from 7SL RNA accounted for 3.1% of the total cellular small RNAs in the control cells, and the abundance dropped about 3.4 folds in Dicer knockdown cells. Dicer-dependent biogenesis of 7SL RNA-derived small RNAs was validated by northern blotting. In vitro cleavage assay using the recombinant human Dicer protein also showed that synthetic 7SL RNA was processed by Dicer into fragments of different lengths. Further functional analysis suggested that 7SL RNA-derived small RNAs do not function like miRNAs, neither do they regulate the expression of 7SL RNA. In conclusion, the current study demonstrated that Dicer can process 7SL RNA, however, the biological significance remains to be elucidated.

MicroRNA-34a inhibits migration and invasion of colon cancer cells via targeting to Fra-1.

MicroRNA-34a (miR-34a), a transcriptional target of p53, is a well-known tumor suppressor gene. Here, we identified Fra-1 as a new target of miR-34a and demonstrated that miR-34a inhibits Fra-1 expression at both protein and messenger RNA levels. In addition, we found that p53 indirectly regulates Fra-1 expression via a miR-34a-dependant manner in colon cancer cells. Overexpression of miR-34a strongly inhibited colon cancer cell migration and invasion, which can be partially rescued by forced expression of the Fra-1 transcript lacking the 3'-untranslated region. The expression of matrix metalloproteinase (MMP)-1 and MMP-9, two enzymes involved in cell migration and invasion, was decreased in miR-34a-transfected cells, and this can be rescued by Fra-1 overexpression. Moreover, we found that miR-34a was downregulated in 25 of 40 (62.5%) colon cancer tissues, as compared with the adjacent normal colon tissues and that the expression of miR-34a was correlated with the DNA-binding activity of p53. Unexpectedly, the DNA-binding activity of p53 was not inversely correlated with Fra-1 expression, and a significant statistical inverse correlation between miR-34a and Fra-1 expression was only observed in 14 of 40 (35%) colon cancer tissues. Taken together, our in vitro data suggest that p53 regulates Fra-1 expression, and eventually cell migration/invasion, via a miR-34a-dependent manner. However, in vivo data indicate that the p53-miR-34a pathway is not the major regulator of Fra-1 expression in human colon cancer tissues.

[L-arginine reduces intestinal epithelial cell apoptosis in rats with severe abdominal infection].

OBJECTIVE: To explore the effect of L-arginine (L-Arg) on intestinal mucosal cell apoptosis in rats with severe abdominal infection. METHODS: Eighteen Wistar rats were randomized into 3 groups, namely the CLP group (n=6) in which the rats were subjected to cecal ligation plus puncture (CLP) to induce severe abdominal infection, L-Arg group (n=6) where the rats received 300 mg/kg peritoneal L-Arg injection following CLP establishment, and the control group (n=6) where the rats underwent ventrotomy only. Intestinal epithelial apoptotic cells were quantified in each group using TUNEL assay 24 h after the operation. RESULTS: Compared with the control group, the rats in CLP and L-Arg groups showed significantly increased number of apoptotic cells in the intestinal epithelium 24 h after the operation (P<0.001). The apoptotic index (AI) in the L-Arg group (18.1-/+2.2) was significantly lower than that in CLP group (20.8-/+2.3, P=0.038). CONCLUSION: Severe abdominal infection results in increased apoptosis of the intestinal epithelial cells in rats, and L-Arg treatment may reduce the cell apoptosis.

[Effect of L-Arginine on intestinal mucosal injury of rats with severe abdominal infection].

OBJECTIVE: To investigate the effect of L-Arginine on intestinal mucosal injury of rats with severe abdominal infection. METHODS: Rats received cecal ligation and puncture (CLP) to reproduce sepsis model. A total of 18 Wistar rats were divided into two groups randomly (each n=9): L-Arginine group and model group. Three hundred mg/kg of L-Arginine was injected into the abdomen in rats of L- Arginine group after CLP. Model group received equal volume of normal saline. Blood sample was harvested and the serum levels of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) were determined at 24 hours after operation in both groups. The histopathological change of intestinal mucosa was observed under light microscope and mucosa damage index was determined. RESULTS: The intestinal mucosal damage was observed both in model group and L- Arginine group after CLP, but the injury was milder in L- Arginine group. There was significant difference in mucosa injury index between L-Arginine group and model group (3.4+/-0.6 vs. 4.1+/-0.5, P<0.05). The serum level of NO [(76.1+/-26.2) micromol/L vs. (87.3+/-16.7) micromol/L, P>0.05] and iNOS [(30.6+/-7.4) U/L vs(44.4+/-6.6) U/L, P<0.01] in L-Arginine group were lower than those in model group. CONCLUSION: L-Arginine could protect against intestinal mucosal injury and depress the serum level of iNOS in severe abdominal infection of rats.