Soil nitrogen availability and microbial carbon use efficiency are dependent more on chemical fertilization than winter drought in a maize-soybean rotation system.

Soil nitrogen (N) availability is one of the limiting factors of crop productivity, and it is strongly influenced by global change and agricultural management practices. However, very few studies have assessed how the winter drought affected soil N availability during the subsequent growing season under chemical fertilization. We conducted a field investigation involving snow removal to simulate winter drought conditions in a Mollisol cropland in Northeast China as part of a 6-year fertilization experiment, and we examined soil physicochemical properties, microbial characteristics, and N availability. Our results demonstrated that chemical fertilization significantly increased soil ammonium and total N availability by 42.9 and 90.3%, respectively; a combined winter drought and fertilization treatment exhibited the highest soil N availability at the end of the growing season. As the growing season continued, the variation in soil N availability was explained more by fertilization than by winter drought. The Mantel test further indicated that soil Olsen-P content and microbial carbon use efficiency (CUE) were significantly related to soil ammonium availability. A microbial community structure explained the largest fraction of the variation in soil nitrate availability. Microbial CUE showed the strongest correlation with soil N availability, followed by soil available C:P and bacteria:fungi ratios under winter drought and chemical fertilization conditions. Overall, we clarified that, despite the weak effect of the winter drought on soil N availability, it cannot be ignored. Our study also identified the important role of soil microorganisms in soil N transformations, even in seasonally snow-covered northern croplands.

Phylogeny and Genetic Divergence among Sorghum Mosaic Virus Isolates Infecting Sugarcane.

Sorghum mosaic virus (SrMV, the genus Potyvirus of the family Potyviridae) is a causal agent of common mosaic in sugarcane and poses a threat to the global sugar industry. In this study, a total of 901 sugarcane leaf samples with mosaic symptom were collected from eight provinces in China and were detected via RT-PCR using a primer pair specific to the SrMV coat protein (CP). These leaf samples included 839 samples from modern cultivars (Saccharum spp. hybrids) and 62 samples from chewing cane (S. officinarum). Among these, 632 out of 901 (70.1%) samples were tested positive for SrMV. The incidences of SrMV infection were 72.3% and 40.3% in modern cultivars and chewing cane, respectively. Phylogenetic analysis showed that all tested SrMV isolates were clustered into three clades consisting of six phylogenetic groups based on 306 CP sequences (this study = 265 and GenBank database = 41). A total of 10 SrMV isolates from South America (the United States and Argentina) along with 106 isolates from China were clustered in group D, while the remaining 190 SrMV isolates from Asia (China and Vietnam) were dispersed in five groups. The SrMV isolates in group F were limited to Yunnan province in China, and those in group A were spread over eight provinces. A significant genetic heterogeneity was elucidated in the nucleotide sequence identities of all SrMV CPs, ranging from 69.0% to 100%. A potential recombination event was postulated among SrMV isolates based on CP sequences. All tested SrMV CPs underwent dominant negative selection. Geographical isolation (South America vs. Asia) and host types (modern cultivars vs. chewing cane) are important factors promoting the genetic differentiation of SrMV populations. Overall, this study contributes to the global understanding of the genetic evolution of SrMV and provides a valuable resource for the epidemiology and management of the mosaic in sugarcane.

TRIM29 facilitates proliferation and malignancy of cholangiocarcinoma cells by activating MAPK and ß-catenin pathways.

BACKGROUND: Tripartite motif-containing 29 (TRIM29) has been found to be involved in the regulation of cancer progression and its function varies depending on the type of cancer. However, the role of TRIM29 in cholangiocarcinoma has yet to be revealed. PURPOSE: This study initially explored the role of TRIM29 in cholangiocarcinoma. METHODS: TRIM29 expression in cholangiocarcinoma cells were scrutinized by quantitative real-time reverse transcription polymerase chain reaction and Western blot. The function of TRIM29 on cholangiocarcinoma cell viability, proliferation, migration and sphere formation abilities were studied by cell count kit-8, clone formation, Transwell and sphere formation assays. TRIM29 effect on the expression of proteins associated with epithelial-mesenchymal transition and cancer stem cell characteristics were researched by Western blot. TRIM29 effect on MAPK and ß-catenin pathway activity was researched through Western blot. RESULTS: TRIM29 was overexpressed in cholangiocarcinoma cells. TRIM29 silencing mitigated the viability, proliferation, migration and sphere formation abilities of cholangiocarcinoma cells, increased E-cadherin expression and decreased the expression of N-cadherin, Vimentin, CD33, Sox2 and Nanog proteins in cholangiocarcinoma cells. The loss of TRIM29 suppressed the expression of p-MEK1/2/MEK1/2 and p-ERK1/2/ERK1/2 in cholangiocarcinoma cells. The inhibition of the MAPK and ß-catenin signaling pathways abrogated the promotion of TRIM29 on cholangiocarcinoma cell viability, proliferation, migration, EMT, and cancer stem cell characteristics. CONCLUSION: TRIM29 plays an oncogenic role in cholangiocarcinoma. It may promote the malignancy of cholangiocarcinoma via inducing the activation of the MAPK and ß-catenin pathways. Thus, TRIM29 may aid in the creation of innovative treatment strategies for cholangiocarcinoma.

High expression of IGHG1 promotes breast cancer malignant development by activating the AKT pathway.

This study researched the exact function of IgG1 heavy chain (IGHG1) on breast cancer (BC) progression. IGHG1 level within BC and paired normal tissues was acquired in Gene Expression Profiling Interactive Analysis dataset. Meanwhile, this work harvested tumor and paired healthy tissues in 42 BC cases. siRNA targeting IGHG1 was transfected into BC cells. SC79 was used to treat the transfected BC cells. CCK-8 assay, clone formation experiment, BrdU assay, Transwell experiment and flow cytometry were carried out to measure the viability, colony formation, proliferation, invasion, and apoptosis of BC cells. Paclitaxel and cisplatin sensitivity of BC cells was evaluated by MTT assay. Real-time quantitative reverse transcription-polymerase chain reaction and Western-blot were performed for measuring mRNA and protein expression. The overexpressed IGHG1 indicated dismal BC survival. IGHG1 silencing attenuated the viability, invasion, proliferation, epithelial-mesenchymal transition, but enhanced the apoptosis of BC cells. IGHG1 silencing enhanced the paclitaxel and cisplatin sensitivity of BC cells. IGHG1 silencing suppressed the activity of the MEK, AKT, and ERK pathways. AKT agonist partially reversed the inhibition of IGHG1 silencing on BC cell malignant phenotype and resistance to paclitaxel and cisplatin. IGHG1 promotes the malignant development of BC by activating the AKT pathway. It may be an effective target for BC treatment.

Microbial network structure, not plant and microbial community diversity, regulates multifunctionality under increased precipitation in a cold steppe.

It is known that the dynamics of multiple ecosystem functions (i. e., multifunctionality) are positively associated with microbial diversity and/or biodiversity. However, how the relationship between microbial species affects ecosystem multifunctionality remains unclear, especially in the case of changes in precipitation patterns. To explore the contribution of biodiversity and microbial co-occurrence networks to multifunctionality, we used rainfall shelters to simulate precipitation enhancement in a cold steppe in Northeast China over two consecutive growing seasons. We showed that an increased 50% precipitation profoundly reduced bacterial diversity and multidiversity, while inter-annual differences in precipitation did not shift microbial diversity, plant diversity, or multidiversity. Our analyses also revealed that increased annual precipitation significantly increased ecosystem, soil, nitrogen, and phosphorous cycle multifunctionality. Neither increased precipitation nor inter-annual differences in precipitation had a significant effect on carbon cycle multifunctionality, probably due to the relatively short period (2 years) of our experiment. The co-occurrence network of bacterial and fungal communities was the most dominant factor affecting multifunctionality, the numbers of negative interactions but not positive interactions were linked to multifunctionality. In particular, our results provided evidence that microbial network topological features are crucial for maintaining ecosystem functions in grassland ecosystems, which should be considered in related studies to accurately predict the responses of ecosystem multifunctionality to predicted changes in precipitation patterns.

SOX12 Promotes Thyroid Cancer Cell Proliferation and Invasion by Regulating the Expression of POU2F1 and POU3F1.

PURPOSE: SOX12 is overexpressed in many cancers, and we aimed to explore the biological function and mechanism of SOX12 in thyroid cancer. MATERIALS AND METHODS: We first analyzed the expression of SOX12 in thyroid cancer using data in The Cancer Genome Atlas. Immunohistochemistry and qRT-PCR were performed to identify SOX12 expression in thyroid cancer tissue and cells. Thyroid cancer cells were transfected with small interfering RNA targeting SOX12, and cellular functional experiments, including CCK8, wound healing, and Transwell assays, were performed. Protein expression was examined by Western blot analysis. A xenograft model was developed to evaluate the effect of SOX12 on tumor growth in vivo. RESULTS: SOX12 expression was increased in thyroid cancer tissue and cells. SOX12 promoted cell proliferation, migration, and invasion and accelerated tumor growth in vivo. The expression of PCNA, Cyclin D1, E-cadherin, Snail, MMP-2, and MMP-9 was affected by SOX12 knockdown. Bioinformatic analysis showed that SOX12 could interact with the POU family. SOX12 knockdown inhibited the expression of POU2F1, POU2F2, POU3F1 and POU3F2, and SOX12 expression showed a positive correlation with POU2F1, POU3F1, and POU3F2 expression in clinical data. POU2F1 and POU3F1 were able to reverse the effect of SOX12 knockdown on thyroid cancer cells. CONCLUSION: SOX12 affects the progression of thyroid cancer by regulating epithelial-mesenchymal transition and interacting with POU2F1 and POU3F1, which may be novel targets for thyroid cancer molecular therapy.

Tumour-suppressing functions of the lncRNA MBNL1-AS1/miR-889-3p/KLF9 axis in human breast cancer cells.

This study aimed to explore the role and potential mechanism of the long non-coding (lncRNA) MBNL1-AS1 in human breast cancer. We included 80 patients with breast cancer in this study. Breast cancer cell lines, including MCF7, SKBR3, MDA-MB-231 and MDA-MB-415, and the normal human breast cell line MCF10A were used in this study. MBNL1-AS1, miR-889-3p mimics, si-Krüppel-like factor 9 (KLF9) or their controls were transfected in the cells. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), Western blotting and immunohistochemistry assay were performed to detect the expression of MBNL1-AS1, miR-889-3p and KLF9. Cell proliferation, invasion and migration were detected. Luciferase reporter gene and pull-down assay were performed to verify the target relationship among MBNL1-AS1, miR-889-3p and KLF9. Glycolysis was also detected after transfection. The expression of the lncRNA MBNL1-AS1 was low in the breast cancer tissues and cells. Lower expression levels of the lncRNA MBNL1-AS1 were associated with poor prognosis of breast cancer. Overexpression of the lncRNA MBNL1-AS1 decreased proliferation, invasion, migration and glycolysis of breast cancer cells. The lncRNA MBNL1-AS1 could interact with miR-889-3p, and KLF9 was the downstream target of miR-889-3p. Moreover, miR-889-3p was negatively correlated with KLF9 and lncRNA MBNL1-AS1. Both miR-889-3p and si-KLF9 could reverse the overexpression of lncRNA MBNL1-AS1 in breast cancer development. The lncRNA MBNL1-AS1 decreased proliferation, invasion, migration and glycolysis of breast cancer via the miR-889-3p/KLF9 axis, which might be a potential biomarker for the diagnosis of breast cancer.

CACNA1B facilitates breast cancer cell growth and migration by regulating cyclin D1 and EMT: the implication of CACNA1B in breast cancer.

PURPOSE: This study mainly aimed to explore the influences of Calcium Voltage-Gated Channel Subunit Alpha1 B (CACNA1B) on the development of breast cancer and the related mechanism. MATERIALS AND METHODS: The information of patients with breast cancer from TCGA database was used for analyses of CACNA1B expression and its prognostic value. Loss- and gain- of functions of CACNA1B were conducted in MCF7 and Bcap-37 cells, respectively. CCK-8, colony formation and transwell assays were applied for evaluating the cell viability and motility. Western blot was used for protein expression detection. RESULTS: We revealed that highly expressed CACNA1B in breast cancer tissues was related to poor prognosis according to the data gained from TCGA database. The outcomes of functional assays showed that depletion of CACNA1B restrained MCF7 cell growth, invasion and migration and high-expression of CACNA1B fortified the growth, invasion and migration in Bcap-37 cells. Finally, we manifested that silencing CACNA1B obviously raised the protein expression level of E-cadherin and reduced the protein levels of Cyclin D1, N-cadherin and Snail in MCF7 cells, whilst, over-expression of CACNA1B reduced the level of E-cadherin and increased the expression of Cyclin D1, N-cadherin and Snail in Bcap-37 cells. CONCLUSIONS: These results identified CACNA1B as a forwarder of the growth, invasion and migration in breast cancer cells.

Risk Factors for Early Recurrence of Gallstones in Patients Undergoing Laparoscopy Combined With Choledochoscopic Lithotomy: A Single-Center Prospective Study.

Objective: For patients with gallstones, laparoscopy combined with choledochoscopic lithotomy is a therapeutic surgical option for preservation rather than the removal of the gallbladder. However, postoperative recurrence of gallstones is a key concern for both patients and surgeons. This prospective study was performed to investigate the risk factors for early postoperative recurrence of gallstones. Methods: The clinical data of 466 patients were collected. Each patient was followed up for up to 2 years. The first follow-up visit occurred 4 months after the operation, and a follow-up visit was carried out every 6 months thereafter. The main goal of each visit was to confirm the presence or absence of gallbladder stones. The factors associated with gallstone recurrence were analyzed by univariate analysis and Cox regression. Results: In total, 466 eligible patients were included in the study, and 438 patients (180 men and 258 women) completed the 2-year postoperative follow-up. The follow-up rate was 94.0%. Recurrence of gallstones was detected in 5.71% (25/438) of the patients. Univariate analysis revealed five risk factors for the recurrence of gallstones. Multivariate Cox regression analysis showed that multiple gallstones, a gallbladder wall thickness of ≥4 mm, and a family history of gallbladder stones were the three predictive factors for postoperative recurrence of gallstones (P < 0.05). Conclusion: The overall 2-year recurrence rate of gallstones after the operation was 5.71%. Multiple gallstones, a gallbladder wall thickness of ≥4 mm, and a family history of gallstones were the three risk factors associated with early postoperative recurrence of gallstones.

Is there a role for growth status in distinguishing gallbladder adenomas from cholesterol polyps? - A retrospective study based on 520 cholecystectomy patients.

INTRODUCTION: Gallbladder adenomas have cancerous potential and occur in 4-8.9% of gallbladder polyps. The growth status (size progression and growth rate during follow-up) of polyps and their effectiveness for predicting adenomas are poorly defined. Herein, we compared adenomas and cholesterol polyps based on the growth status and evaluated the reported risk factors in predicting neoplasm. METHODS: We enrolled 520 patients who underwent preoperative ultrasonic follow-up more than 6 months with post-cholecystectomy pathologically confirmed gallbladder polyps. The patients were classified into adenoma and cholesterol polyp groups. Growth status, clinical characteristics, laboratory data, ultrasonic findings were reviewed and compared between the groups. RESULTS: Seventy-nine adenomas and 441 cholesterol polyps were analyzed. The mean diameter of adenomas (cholesterol polyps) was 7.24 ± 4.36 mm (6.23 ± 2.88 mm) in the initial and 12.06 ± 4.61 mm (10.05 ± 2.95 mm) in the preoperative examination. The median size progression (range) of polyps in the cholesterol polyps [3 (0, 22)] mm was smaller than that in adenomas [4 (0, 21)] mm (p = .075). The mean growth rate of adenomas (1.07 ± 1.33 mm/6 months) was slightly faster than in cholesterol polyps (0.83 ± 1.04 mm/6 months) (p = .338). The indicators significantly associated with adenomas were age >49.5 years, lack of gallbladder polyps/cholelithiasis family history, polyp size >11.5 mm and solitary polyp (p = .005, p = .027, p = .001, and p = .021, respectively). CONCLUSION: Growth status was not a valuable modality to distinguishing gallbladder adenomas from cholesterol polyps. Risk factors such as age, polyp size, and solitary polyp were effective in predicting adenomas.

Comparative genomics reveals insights into genetic variability and molecular evolution among sugarcane yellow leaf virus populations.

Yellow leaf disease caused by sugarcane yellow leaf virus (SCYLV) is one of the most prevalent diseases worldwide. In this study, six near-complete genome sequences of SCYLV were determined to be 5775-5881 bp in length. Phylogenetic analysis revealed that the two SCYLV isolates from Réunion Island, France, and four from China were clustered into REU and CUB genotypes, respectively, based on 50 genomic sequences (this study = 6, GenBank = 44). Meanwhile, all 50 isolates were clustered into three phylogroups (G1-G3). Twelve significant recombinant events occurred in intra- and inter-phylogroups between geographical origins and host crops. Most recombinant hotspots were distributed in coat protein read-through protein (RTD), followed by ORF0 (P0) and ORF1 (P1). High genetic divergences of 12.4% for genomic sequences and 6.0-24.9% for individual genes were determined at nucleotide levels. The highest nucleotide diversity (π) was found in P0, followed by P1 and RdRP. In addition, purifying selection was a main factor restricting variability in SCYLV populations. Infrequent gene flow between Africa and the two subpopulations (Asia and America) were found, whereas frequent gene flow between Asia and America subpopulations was observed. Taken together, our findings facilitate understanding of genetic diversity and evolutionary dynamics of SCYLV.

lncRNA FLVCR1-AS1 regulates cell proliferation, migration and invasion by sponging miR-485-5p in human cholangiocarcinoma.

Long non-coding RNA (lncRNA) FLVCR1 antisense RNA 1 (FLVCR1-AS1) serves a crucial role in many types of cancer; however, to the best of our knowledge, the biological effect of FLVCR1-AS1 in cholangiocarcinoma (CCA) remains unclear. The present study aimed to elucidate the involvement of FLVCR1-AS1 in the regulation of human CCA cell growth, migration and invasion, as well as the mechanisms underlying its effect. The expression levels of FLVCR1-AS1 in CCA tumor tissues, adjacent normal tissues, CCA cell lines and a cholangiocyte cell line were determined by reverse transcription-quantitative polymerase chain reaction. A significantly higher expression level of FLVCR1-AS1 was identified in CCA tumor tissues and the CCA cell lines HuCCT1 and CCLP1 compared with the normal controls. Short hairpin RNA targeting FLVCR1-AS1 (shFLVCR1-AS1) and a control plasmid (shNC) were transfected into CCA cell lines. Cell proliferation, colony formation, migration and invasion of CCA cells transfected with shFLVCR1-AS1 were significantly suppressed compared with the shNC groups. The expression levels of migration and invasion-associated proteins, including Twist, matrix metalloproteinase (MMP)-2 and MMP-9, were also significantly suppressed by shFLVCR1-AS1-treatment. Furthermore, FLVCR1-AS1 knockdown inhibited tumor growth in a xenograft model. Mechanistically, FLVCR1-AS1 was demonstrated to sponge microRNA-485-5p (miR-485-5p) in human CCA. The expression of miR-458-5p was significantly decreased in CCA tissue compared with normal tissue, and Pearson's correlation analysis revealed that FLVCR1-AS1 expression was negatively correlated with miR-485-5p expression in CCA tissues. These results suggested that lncRNA FLVCR1-AS1 may be used as a novel therapeutic target and a potential diagnostic marker for CCA.