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Objective: To observe the effect of RND3 intervention on injury of pancreatic acinar cells AR42J induced by caerulein in rats, and explore regulatory mechanism of RND3 in acute pancreatitis (AP) . Methods: Lentiviral AP models with RND3 gene silencing and overexpression, and AR42J cell AP models induced by caerulein were constructed in 6 groups (NC control group, caerulein treatment group, si-con negative control group, pcDNA negative control group, si-RND3 silencing group, and pcDNA-RND3 overexpression group) . After the administration of caerulein, mRNA and protein expression level of RND3 in AR42J cells were quantified at time points of 0 h, 4 h, 8 h, 12 h and 24 h by qRT-PCR and Western blot, respectively. Cell survival and LDH leakage were detected by CCK-8 method. Apoptosis rate of AR42J cells was detected by flow cytometry. mRNA expression levels of related inflammatory factors (TNF-α, IL-6, IL-1β and IL-18) were detected by qRT-PCR. Western blot was used to detect expressions of apoptosisrelated proteins (GSDMD, NLRP3, Cleaved caspase-1 and caspase-1) in AR42J cells. Results: Expression of RND3 mRNA and protein in AR42J cells induced by caerulein was inhibited, and expression of RND3 was inhibited more significantly with the increase of caerulein treatment time. Compared with NC control group, expressions of TNF-α, IL-6, IL-1β and IL-18 in AR42J cells of rats in si- RND3 group and caerulein treatment group were up-regulated (P<0.05), at the same time, expressions of apoptosis-related proteins GSDMD, NLRP3, Cleaved caspase-1 and caspase-1 were increased. On the contrary, levels of cell inflammation and apoptosis were significantly reduced in pcDNA-RND3 overexpression group (P<0.05) . Conclusion: RND3 can reduce the inflammatory reaction and apoptosis of rats pancreatic acinar cells AR42J, which mechanism is related to the activity of NLRP3/caspase-1/GSDMD pathway. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Contemporary Pediatrics is the property of Xiangya Medical Periodical Press and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: To explore the effect of methyltransferase METTL3 on the proliferation, migration and osteogenic differentiation of jaw bone marrow mesenchymal stem cells (JBMMSCs). Methods: JBMMSCs were cultured in vitro and transfected with lentivirus to construct a cell model of silencing METTL3 gene. After transfection, the mRNA level (P<0.001) and protein level (P<0.01) of JBMMSCs in the siMETTL3 group decreased. The proliferation, migration, alkaline phosphatase activity, calcium deposition, and osteogenesis-related factors osteocalcin(OCN)mRNA and protein level of the two groups of JBMMSCs were detected. Results: Compared with the siNC group, the proliferation activity of the siMETTL3 group decreased after 24, 48, 72, and 96 h of culture (all P<0.001), and the cell migration ability decreased after 24 h (P<0.05). The activity of alkaline phosphatase in the siMETTL3 group decreased after 7 days of osteogenic induction, and the mineralization deposition after 21 days was less than that in the siNC group. qRT-PCR and Western blot showed that the mRNA level(P<0.001) and protein expression level(P<0.05) of OCN reduced. Conclusion: Silencing METTL3 can inhibit the proliferation, migration and osteogenic differentiation of JBMMSCs. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of angiopoietin 4 (ANGPT4) on the odontogenic differentiation of human dental pulp stem cells. Methods This study has been reviewed and approved by the Ethics Committee, and informed consent has been obtained from patients. Human premolars were fixed, decalcified, dehydrated, embedded, and sectioned. Immunofluorescence staining was used to observe the expression and localization of ANGPT4. Human dental pulp stem cells (hDPSCs) were isolated and cultured in vitro. The growth state and morphology of hDPSCs were observed under an inverted phase contrast microscope. The expression of cell surface-related molecular markers was detected by flow cytometry. Alkaline phosphatase and alizarin red S staining were used to detect the odontogenic differentiation potential of hDPSCs. Oil-red O staining was used to detect the adipogenic differentiation potential of hDPSCs. RNA was extracted from hDPSCs at different time points after odontogenic induction, and RT-qPCR was used to analyze the mRNA expression of ANGPT4 and odontogenic-related genes during the odontogenic differentiation of hDPSCs in vitro. siRNA gene silencing technology was used to silence the expression of ANGPT4 in hDPSCs, and the silencing efficiency was detected by RT-qPCR and Western Blot. After silencing ANGPT4 in hDPSCs for 24 h, odontogenic induction was performed. Alkaline phosphatase and alizarin red S staining were performed on the 7th and 14th of induction to detect the odontogenic differentiation ability of hDPSCs after silencing ANGPT4. Results Immunofluorescence staining of human premolars showed that ANGPT4 was expressed in odontoblasts and sub-odontoblastic cell-rich zone. hDPSCs were in good condition after 14 days of isolation and culture. Under the microscope, multiple cell colonies were observed, and the cell morphology was uniform and long spindle-shaped. The results of flow cytometry showed that hDPSCs expressed mesenchymal stem cell markers CD105 (90.42%) and CD90 (97.15%), but did not express hematopoietic cell markers CD45 (0.01%) and CD34 (0.08%). After odontogenic and adipogenic induction of hDPSCs, alkaline phosphatase staining, alizarin red S staining and oil red O staining were positive. The results of RT-qPCR after the odontogenic induction of hDPSCs showed that ANGPT4 was highly expressed on the 5th, 7th, 11th and 14th days of differentiation of hDPSCs (P

Tomato (Solanum lycopersicum) is one of the most popular vegetables worldwide and is a classic model plant for studying fruit development and ripening due to its short growth cycle, clear genetic background and ease of molecular manipulation. This paper used virus-induced gene silencing (VIGS) to construct SlWRKY53b gene-silenced tomato fruits and analyzed the effect of SlWRKY53b gene silencing in the tomato fruit ripening process. We found that transient silencing of SlWRKY53b resulted indelayed inbroken color, higher chlorophyll contents (P<0.05) and reduced carotenoid contents (P<0.05) in tomato fruits, and color difference results indicated that the differences in L*, a* and b* values were consistent with fruit color changes. Further studies showed that genes significantly down-regulated (P<0.01) in SlWRKY53b gene-silenced tomato fruits include the chlorophyll degradation-related genes (NYC1, PAO, PPH, SGR1), carotenoid synthesis-related genes (PSY1, PDS, ZDS), ethylene synthesis pathway-related genes (ACO1, ACS2, NOR, ACO3, E4, RIN), and cell wall degradation-related genes (PG, EXP, CEL2). Correlation analysis showed that the expression of SlWRKY53b was negatively correlated with chlorophyll contents and positively correlated with carotenoid contents and the expression of maturation-related genes. These results suggest that inhibition of SlWRKY53b expression at the transcriptional level can achieve the effect of delaying tomato fruit ripening, indicating that SlWRKY53b plays a role as a facilitator in the tomato fruit ripening process. [ABSTRACT FROM AUTHOR]

Tomato (Solanum lycopersicum), the fruit is rich in vitamins, lycopene and other beneficial nutrients to human body, is one of the important vegetable crops. ERF is a type of plant-specific transcription factor, which is an ethylene response factor and can directly regulate the expression of target genes. However, the function of SlERF14 in tomato remains unclear. In this paper,31 ERF gene families in tomato, Arabidopsis thaliana and sweet potato were analyzed by phylogenetic tree, and five ERF genes from a clade with high homology of SlERF14 were screened and summarized for amino acid sequence alignment. The results showed that the function of these genes may be more conservative, and the gene properties and structure are more similar. The expression of SlERF14 gene in tomato was analyzed, and it was found that SlERF14 gene might play a role in fruit ripening. In addition, by virus-induced gene silencing technique (VIGS), it was found that SlERF14 gene silencing promoted the ripening of tomato fruits, resulting in decreased chlorophyll content and increased carotenoid content. After silencing for 15 days, chlorophyll a content was significantly down-regulated (P<0.01), and carotenoid content was significantly up-regulated (P<0.01). The expression levels of carotenoid synthesis related genes (PSY1, PDS, ZDS), chlorophyll degradation related genes (NYC1, PAO, PPH, SGR1), and ethylene anabolic pathway related genes (ACO1, ACO3, ACS2, RIN) all showed an up-regulated trend (P<0.05). It is concluded that SlERF14 gene plays a negative regulatory role in tomato ripening and senescence. In conclusion, the bioinformatics analysis of SlERF14 gene family and its function were studied in this paper, which is of great significance to further study the function of SlERF14 gene in tomato. [ABSTRACT FROM AUTHOR]

BACKGROUND: It has been reported that makorin ring finger protein 1 (MKRN1) knockdown can inhibit the growth of tumor cells, but whether MKRN1 plays a role in the body as a tumor-related protein needs further study. The siRNA-mediated gene silencing is transient, and therefore, the duration is short and the conventional lentivirus technique also has an off-target effect. A cell line stably silencing MKRN1 gene constructed using Tet-on lentivirus system has reversible advantages and can avoid the above shortcomings, which is of great significance for the further study on the biological function of MKRN1. OBJECTIVE: To construct a MKRN1 gene silencing vector mediated by the Tet-on lentivirus system in order to study the effect of MKRN1 gene silencing on the proliferation and apoptosis of human embryonic kidney cells HEK-293T. METHODS: Using RNAi technique, three short hairpin RNAs (shRNAs) were designed for MKRN1 gene, namely sh1, sh2, and sh3, and three shRNA sequences were ligated into tetracycline-induced expression vector pTripz. The correct sequence was verified by sequencing. HEK-293T cells were transfected with the three-plasmid system and cultured in the complete medium containing 1 mg/L doxycycline for lentivirus packaging. The virus solution was collected at 48 and 72 hours and concentrated to infect HEK-293T cells. Forty-eight hours after infection, the virus infection efficiency was observed by fluorescence microscope. The knockout efficiency of MKRN1 was detected by real-time fluorescence quantitative PCR and western blot assay. The successful construction of tetracyclineinduced expression system was detected by western blot assay. Colony formation test and western blot assay were used to detect the effect of silencing MKRN1 expression on the proliferation and apoptosis of HEK-293T cells. RESULTS AND CONCLUSION: (1) Three MKRN1-shRNAs, named sh1, sh2, and sh3, were successfully inserted into the Tet-on vector, and the sequence of each vector was correct. HEK-293T cells were successfully transfected with lentivirus. (2) RT-qPCR and western blot results indicated that the MKRN1 knockdown level of the first sequence of the three shRNA sequences was significantly decreased (P < 0.05), and the expression of the target gene was rescued in the silent group without doxycycline induction. (3) The proliferation of HEK-293T cells decreased after MKRN1 silencing detected by colony formation experiments (P < 0.05). Western blot assay results revealed that after MKRN1 knockdown, the expression of proliferating cell nuclear antigen was down-regulated, the expression of BAX was increased, the expression of Bcl2 was down-regulated, and the Bcl2/BAX ratio was down-regulated (P < 0.001). (4) To conclude, the HEK-293T cell line stably silencing MKRN1 can be constructed by using Tet-on lentiviral vector system, and the silencing of MKRN1 can inhibit the proliferation of HEK293T cells and promote their apoptosis. The construction of stably silencing MKRN1 HEK-293T cells can lay a foundation for the further study on the biological function of MKRN1 and also provide a methodological reference for the study of the biological function of other genes. [ABSTRACT FROM AUTHOR]

AIM: To examine the impact of inhibiting the expression of the tetraspanin 1 (Tspan1) gene on the resistance of colorectal cancer (CRC) cells to oxaliplatin (OXA), as well as to elucidate the underlying mechanism. METHODS: This study involved the collection of cancer tissues from two groups of patients with CRC: one group comprising 31 patients who exhibited sensitivity to OXA chemotherapy (referred to as the OXA sensitive group), and the other group comprising 26 patients who showed resistance to OXA chemotherapy (referred to as the OXA resistant group). The mRNA and protein expression levels of Tspan1 in the cancer tissues were measured using quantitative reverse transcription)polymerase chain reaction (RT-qPCR) and Western blot analysis. To establish an OXA-resistant cell line, HCT116/ OXA, the concentration of OXA was incrementally increased. The proliferation activity of the drug-resistant cell lines was assessed using the MTT assay, and the half inhibitory concentration (IC50) and drug resistance index (RI) were calculated. The mRNA and protein expression levels of Tspan1 were measured in both the drug-resistant HCT116/OXA cells and the parent HCT116 cells using RT-qPCR and Western blot analysis. Small interfering RNA (siRNA) technology was employed to silence the expression of the Tspan1 gene in HCT116/OXA cells, and the mRNA and protein expression levels of Tspan1 were measured in the transfected drug-resistant cell lines. Various concentrations of OXA were administered to the transfected drug-resistant cell lines, and the MTT assay was employed to assess cell proliferation activity and calculate the IC50 value. Cell apoptosis was examined using flow cytometry. The immunofluorescence intensity of the autophagy index LC3B was determined using an immunofluorescence assay, and the expression levels of autophagy-related proteins such as LC3, beclin-1, and P62 were evaluated using Western blot analysis. RESULTS: In comparison to the OXA sensitive group, the expression levels of Tspan1 mRNA and protein in the cancer tissues of patients in the OXA resistant group were significantly elevated (P<0. 01). Likewise, the HCT116/OXA cells demonstrated higher mRNA and protein expression levels of Tspan1 compared to the HCT116 cells (P<0. 01). The IC50 values of the drug-resistant cell line HCT116/OXA and its parent HCT116 cells were (101. 6±2. 7) and (16. 0±0. 5) mg/L, respectively, resulting in an RI value of 6. 4 after 48 h of treatment with various OXA concentrations. The mRNA and protein expression levels of Tspan1 in the drug-resistant HCT116/OXA cell line was significantly greater than that in the parent HCT116 cell line (P<0. 01). Silencing the expression of the Tspan1 gene significantly reduced the mRNA and protein expression levels of Tspan1 and reversed the drug resistance to OXA in HCT116/OXA cells (P<0. 05). It also increased the rate of apoptosis in HCT116/OXA cells when exposed to OXA (P<0. 05). Additionally, it led to a decrease in the fluorescence intensity of the LC3B protein, a downregulation of the LC3-II/LC3-I protein ratio and the expression level of beclin-1 protein in cells (P<0. 05), and an increase in the expression of the P62 protein (P<0. 05). CONCLUSION: Inhibiting autophagy through the silencing of Tspan1 gene expression effectively reversed OXA resistance in CRC cells. [ABSTRACT FROM AUTHOR]

Phosphorylation, a widespread post-translational modification observed in eukaryotic cells, plays a crucial role in regulating various metabolic activities and cell signal transduction processes. In organisms, plant viral proteins have the ability to modulate the biological activity of proteins through phosphorylation and dephosphorylation modifications, thereby controlling the transmission of biological signals within cells. The phosphorylation-modified plant viral proteins actively participate in regulating viral infection, viral RNA synthesis, RNA silencing and host gene expression. In recent years, extensive research has been conducted to unravel the molecular mechanisms underlying the phosphorylation of plant virus proteins. This article provides an overview of phosphorylation modification sites, methods for identifying phosphorylation, and the biological functions of plant viral proteins, aiming to offer valuable insights into the role of phosphorylation in plant-virus interactions. [ABSTRACT FROM AUTHOR]

Virus-induced gene silencing（VIGS）technology has been widely used in plant gene function research. The efficiency of gene silencing in soybean mediated by tobacco rattle virus（TRV）-based vector system remains to be clarified. The seamless cloning technique was used to construct a TRV-VIGS system to explore the silencing efficiency in soybean target genes in different tissues with different inoculation methods, providing a basis for gene function studies in soybean. Taken phytoene desaturase（GmPDS）and ubiquitin ligase（GmATL3）as target genes, pTRV1 and the recombinant vectors solution were inoculated into soybean（Glycine max）plants Zhonghuang13 by three methods: injection, agroinoculation and a combination of injection plus agroinoculation. The silencing phenotypes were observed at 28 d after inoculation, and the relative expressions of the genes in the roots and leaves were quantified by RT-qPCR to determine the silencing effectiveness of the different methods. Silencing of GmPDS resulted in yellowing and chlorosis on the edges and inside of the leaves, while chlorosis spots and fold chlorosis phenotypes in the leaves surface occurred. The results of RT-qPCR showed that silencing efficiency of GmPDS of all three inoculation methods was close to 100%. The silencing efficiency of GmATL3 by the injection was 80%-95% in the leaves and 40%-60% in the roots; the silencing efficiency of the root inoculation and injection plus root inoculation was 70%-90% in roots and 15%-50% in leaves. Different inoculation methods caused different levels of silencing phenotypes and silencing efficiencies for different endogenous genes. The silencing efficiency of an inoculation method on different tissues was different, and the highest silencing efficiency on the leaves and roots were injection method and injection plus agroinoculation, respectively. [ABSTRACT FROM AUTHOR]

The use of RNA interference (RNAi) is becoming a routine method of scientific discovery and treatment of human diseases. However, its application is hindered by adverse effects, especially the off-target effects. In this work, we established an shRNA test platform based on the plasmid to test the silencing ability of short hairpin RNA (shRNA) targeting survivin gene designed according to shRNA online software and then screened out two shRNAs with excellent silencing ability. The off-target effects were measured by testing the silencing ability of two selected shRNAs against targets with single-nucleotide mutations at different locations. Based on our understanding of the 3′ tail function of microRNA (miRNA) and small interfering RNA (siRNA), we propose a method to reduce the off-target effect of short hairpin RNA by shortening the complementary length of the 3′ tail and target sequence of siRNA only. This method can reduce the off-target effect of shRNA without damaging the silencing efficiency of shRNA gene, so as to effectively improve the specificity of shRNA gene silencing. Compared to previous studies of the weak base pairing of “seed” and 3′ regions to reduce the off-target effect, our method is simpler and more efficient, without being limited by the sequence of 3′ regions of the antisense strand. This method of reducing the off-target effects provides new ideas for the design of shRNA and siRNA, simplifies the sequence restriction in the design of shRNA and siRNA drugs to a certain extent, and broadens their use and prospects as therapeutic and diagnostic tools in medical treatment. [ABSTRACT FROM AUTHOR]

Copyright of West China Journal of Stomatology is the property of Sichuan University, West China College of Stomatology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Modern Laboratory Medicine is the property of Journal of Modern Laboratory Medicine Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Applied Entomology is the property of Chinese Journal of Applied Entomology, Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: To establish MDA-MB-231/cisplatin (DDP) resistant cell lines of triple-negative breast cancer, and to explore the mechanism of transforming growth factor β1 (TGF-β1) regulating DDP resistance of triple-negative breast cancer. Methods:MDA-MB-231 resistant cell lines (MDA-MB-231/DDP) were established by low-dose intermittent induction method. TGF-β1 silencing cells were constructed in MDA-MB-231/DDP and they were divided into TGF-β1 silencing group (sh-TGF-β1), negative control group and control group. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect TGF-β1 content. MDA-MB-231 cells and MDA-MB-231/DDP cells were divided into MDA-MB-231 group (MDA-MB-231 sensitive cells cultured normally) and MDA-MB-231-DDP group (MDA-MB-231 sensitive cells cultured normally DDP resistant cells), TGF β1-shRNA group (MDA-MB-231DDP cells transfected with TGFβ1-shRNA lentiviral vector) and MDA-MB-231-DDP combined with 3-MA group (MDA-MB-231 DDP cells were treated with 5 m M 3-MA for 2 h). Half inhibitory concentration (IC50) of drug-resistant strains was detected by cell counting kit (CCK-8), and drug resistance index and reverse resistance index were calculated. TGF-β1 content was detected by RT-qPCR, and expression levels of TGF-β1 and autophagy related proteins LC3-I and LC3-II were detected by Western blot. The changes of autophagy flow were observed by laser confocal microscopy. SPSS 20.0 software was used for statistical analysis. Results: The DDP resistant cell line MDA-MB-231/DDP was established successfully, and the resistance index was 5.231. The m RNA expression and protein expression of TGF-β1 in the MDA-MB-231/DDP cells were significantly up-regulated compared with MDA-MB-231 cells (P＜0.05). The expression of autophagy related protein LC3-II/LC3-I in the MDA-MB-231/DDP resistant cells were significantly higher than those in the MDA-MB-231 cells (P＜0.05). The expression of autophagy related protein LC3-II/LC3-I in the MDA-MB-231/DDP cells was significantly decreased after the application of autophagy inhibitor 3-methyladenine (3-MA) (P＜0.05). After silencing TGF-β1 gene in the MDAMB-231/DDP cells, the drug resistance index in the DDP resistant cell lines decreased from 5.231 to 3.404, while the expression of autophagy related protein LC3-II/LC3-I were decreased (P＜0.05), and the yellow and red spots were significantly reduced by confocal laser microscopy, it showed that autophagy was inhibited. Conclusion: TGF-β1 is associated with DDP resistance in the triple-negative breast cancer, and the mechanism may be increased autophagy causing DDP resistance in the MDA-MB-231 cells. By silencing TGF-β1, autophagy levels are reduced and DDP sensitivity of triple-negative breast cancer cells is restored. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Northwest A & F University - Natural Science Edition is the property of Editorial Department of Journal of Northwest A&F University (Natural Science Edition) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective To investigate the effects of over expression and low expression of antisense transcripts of circular RNA cerebellar degeneration associated protein 1 (CDR1as) in Balb/C mouse bone marrow mesenchymal stem cells (BMSCs) on factors related to osteogenesis and angiogenesis. Methods BMSCs were cultured and identified in vitro. The lentiviral (LV) vector containing the overexpressed and silenced circRNA CDR1as genes and the control lentivirus were respectively transfected into mouse BMSCs, and stable cell lines were screened. The cells were divided into the circRNACDR1as over expression group and the over expression control group, and the CircRNACDR1as low expression group and the low expression control group. The components were stained with Alizarin Red S and alkaline phosphatase after 14 and 21 days of osteoinduction; qRT-PCR was used to detect the target genes circRNA CDR1as, osteogenic differentiation markers alkaline phosphatase (ALP), runt- related transcription factor 2 (RUNX2), osteocalcin (OCN), osteopontin (OPN), osterix(Osx), collagen I (COL-1), and the mRNA expression levels of vascular endothelial grown factor (VEGF) and angiogenin-1 (Ang-1). Results The results of alizarin red staining and alkaline phosphatase staining showed that the extracellular matrix calcium precipitation and ALP staining area of the over expression experimental group was greater than its control group, and those of the low expression experimental group was less than its control group. As the number of days of osteogenic induction increased, the calcium precipitation and ALP staining in each group also increased. RT-PCR results showed that the mRNA expression levels of circRNA CDR1as, ALP, RUNX2, OCN, OPN, OSX, COL-1, VEGF and Ang-1 in the over expression experimental group BMSCs were significantly increased (P

RNA interference (RNAi) is a highly conserved gene silencing phenomenon in eukaryotes, which has shown broad potential application in medicine and plant protection. Recently, RNAi-related products have been or about to been in the pharmaceutical and pesticide markets. For RNAi-based fungicides, although researchers have made a lot of efforts in the development of its products, the commercial application of RNAi against plant pathogens has not arrived on the market yet. Here, the development of RNAi research since 1990 is summarized, and new ideas that may help improve the development of RNAi-based fungicides are introduced. At the same time, the influence of RNAi-based fungicides on traditional chemical fungicides is discussed, which is the innovation and application of RNAi fungicides. [ABSTRACT FROM AUTHOR]

【Objective】Tomato zonate spot orthosporospirus (TZSV) are transmitted in a persistent way by thrips, which has seriously affected the yield and quality of important vegetables and ornamental crops in Southwest China, and has brought serious harm to local agricultural production. The N and NSm genes of TZSV are closely related to virus classification, movement and systemic infection. The study aims to provide materials for further research on the function of TZSV N and NSm genes in virus infection, and provide some basis for disease resistance breeding and green control in the field based on constructing N and NSm gene silencing vectors. 【Method】In the study, specific primers were designed according to the sequences of TZSV N and NSm genes, N and NSm gene fragments were amplified by RT-RCR and were cloned into the vector of pEASY-Blunt-Zero. The pEASY-Blunt-Zero vector containing N and NSm gene sequences and pTRV-pTV00 vector were both digested with Bam HI and Hind III restriction enzymes and were purified and recycled, thereafter the purified products were linked to pTRV-pTV00 vector using T4 DNA ligase to obtain pTRV-PTV00-N and pTRV-PTV00-NSm recombinant vectors, which were then transferred into Agrobacterium tumefaciens GV3101 and injected into Nicotiana benthamiana and N. tabacum cv. K326. The silencing efficiency of N and NSm genes was detected by real-time fluorescent quantitative PCR (RT-qPCR), and the content of N and NSm proteins was detected by indirect enzyme-linked immunosorbent assay (ID-ELISA). 【Result】RT-qPCR analysis showed that the silencing efficiency of N and NSm genes in N. benthamiana and N. tabacum cv. K326 was greater than 90% 5 days after TZSV inoculation. The results of ID-ELISA showed that the content of two proteins decreased significantly in different time, and the content of NSm protein decreased significantly for 9 consecutive days. 【Conclusion】The construction of N and NSm gene silencing vectors was successful. The experiment was successfully conducted on N. benthamiana and N. tabacum cv. K326.The construction of N and NSm gene silencing vectors provides materials for studying the pathogenesis of TZSV, and provides a theoretical basis for breeding and control of TZSV resistance in the field. [ABSTRACT FROM AUTHOR]

[Objectives] To clone the Hyphantria cunea glutathione S-transferases (GST) gene (HcGST-E1 ) and verify the role of this gene in the response of H. cunea to quercetin, a common secondary substance in host plants. [Methods] HcGST-E1 was cloned based on the quercetin-induced midgut transcriptome of H. cunea and real-time, quantitative PCR results. The gene was then silenced using RNAi technology and the effect of this on the weight and survival of quercetin-treated 4th instar larvae investigated. [Results] The HcGST-E1 was successfully cloned and silenced. dsHcGST-E1 effectively inhibited the expression of HcGST-E1 with a maximum silencing efficiency of 62.3%. Over the same period of time, 1 000 ng of dsHcGST-E1 was more effective than 500 ng dsHcGST-E1. The survival rates of HcGST-E1 silenced larvae that had been fed an artificial diet containing either 0.5% or 1.0% quercetin were 40.00% and 33.33%, respectively, significantly lower than their respective control groups (73.33% and 66.67%; P < 0.05) or blank control groups (86.67% and 80.00%; P < 0.05) . The body weight of HcGST-E1 silenced H. cunea larvae did not fluctuate significantly, and their weight was lower than that of larvae in both respective, and blank, control groups. Larvae in the 5% quercetin treatment group were not significantly lighter than those in the respective control group, but were significantly lighter than those in the blank control groups on day 6 (P < 0.01). Similarly, larvae in the 1.0% quercetin treatment group were not significantly lighter than those in the respective control group, but were significantly lighter than those in the blank control groups on days 4 and 6 (P <0.05). [Conclusion] Silencing the HcGST-E1 gene increased the susceptibility of H. cunea larvae to quercetin, retarding their growth and development. This indicates that HcGST-E1 plays a role in the detoxification and metabolization of quercetin. These results provide a basis for further research on the role of GST genes in the resistance of H. cunea to plant secondary compounds. [ABSTRACT FROM AUTHOR]

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Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Henan University of Technology Natural Science Edition is the property of Henan University of Technology Journal Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agronomica Sinica is the property of Crop Science Society of China and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The WRKY transcription factor gene family is a plant-specific transcription factor that plays important roles defense responses. Studies in model plant Arabidopsis demonstrated that WRKYs function downstream of mitogen activated-protein kinase (MAPK) signaling cascade and participate in defense responses through activating the expression of defense-related genes. However, the roles of WRKYs in defense responses have not been previously investigated in paleopolyploidy soybean. Bioinfomatic analysis revealed that there are three pair of GmWRKY33 genes in the soybean genome. The identity of first two pair of GmWRKY33 genes is greater than 84% (named as GmWRKY33A ). The identity of genes within the same pair is greater than 95%. A 300 bp fragment highly homologous to these four GmWRKY33A was chosen to clone into bean pod mosaic virus (BPMV)-based silencing vector (BPMV-VIGS) to achieve the goal of silencing four GmWRKY33A genes simultaneously. In this study, we simultaneously silenced four homologous genes of GmWRKY33A using a bean pod mottle virus (BPMV) vector carrying a single fragment of GmWRKY33A . Comparing the silenced plants with the vector control plants, no evident morphological phenotypes were observed. However, the GmWRKY33A- silenced plants exhibited significantly reduced resistance to Pseudomonas syringae pv. glycinea ( Psg ), Xanthomonas axonopodis pv. glycine ( Xag ), as well as to soybean mosaic virus (SMV). Furthermore, we demonstrated that silencing these GmWRKY33A genes significantly inhibited the activation of Gm MPK3/ Gm MPK6 induced by Psg infection. Collectively, our results suggest that Gm WRKY33As are involved in soybean immunity through regulating the transcription of GmMPK3/6 genes or activating the kinase activities of Gm MPK3/6. Taken together, our results demonstrated that Gm WRKY33As are positive regulators of soybean immune responses.

To clarify the roles of the heat shock protein gene Hsp70 in the sclerotial formation and pathogenicity of Sclerotinia sclerotiorum , we employed reverse transcription PCR (RT-PCR) to clone Hsp70 from S . sclerotiorum and performed sequence analysis. Quantitative real-time PCR (qRT-PCR) was employed to determine the relative expression levels of Hsp70 at different growth stages and under the stress induced by cyclic adenosine monophosphate (cAMP) and low and high temperatures. The thermal stability of Hsp70 was measured. The Agrobacterium -mediated method was employed to construct the Hsp70 -silenced strain. The pathogenicity and fungicide resistance of strains were tested by inoculation in detached rapeseed leaves and cultivation in the media containing procymidone and thiophanate-methyl, respectively. The results showed that the cloned Hsp70 had a total length of 1 890 bp and close relationship with the Hsp70 gene of Ciborinia . Hsp70 showcased the highest expression level in sclerotia, which was more than 30 times higher than that in hyphae. The cAMP stress significantly induced the expression of Hsp70 . The expression level of Hsp70 showed an increasing-decreasing-increasing trend at 40 ℃ and no significant change at 4 ℃. Recombinant strain with high expression of Hsp70 showed good thermal stability. The Hsp70- silenced transformant did not form sclerotia, with decreased pathogenicity and fungicide resistance. This study reveals that Hsp70 plays an important role in the sclerotial formation and stress resistance of S . sclerotium , providing reference for further in-depth research on the biological roles of Hsp70 in S . sclerotium .

Copyright of Chinese Journal of Applied Entomology is the property of Chinese Journal of Applied Entomology, Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: To investigate the effect of suppressor of cytokine signaling (SOCS1) gene silencing of dendritic cells (DCs) on the immune effect of cervical cancer cells in the tumor bearing mouse model of cervical cancer cell TC-1. Methods: The tumor bearing mouse model of cervical cancer cell line TC-1 was constructed. The lentivirus vector containing SOCS1 silencing gene was infected with DCS cells. After cellular immunization of tumor bearing mouse, the tumor growth and survival rate of mouse were monitored, the cleavage rate of spleen cells to TC-1 cells in vitro and the expression of γinterferon (IFN-γ) and interleukin-12 (IL-12) were detected. Results: DCs SOCS1 gene silencing can prolonged the survival of mouse, enhanced the inhibitory effect of DCs on tumor growth in vivo, enhanced the lysis rate of mouse spleen cells to TC-1 cells in vitro (P<0.05), increased the expression of IL-12 factor in serum of mouse (P<0.05) and IFN-γexpression of mouse spleen cells (P<0.05). However, the number of effector cells in spleen of mouse was not affected (P>0.05). Conclusion: In vivo experiments in mouse show that DCs SOCS1 gene silencing could enhance the killing effect of effector cells on tumor cells in mouse to a certain extent. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Oil Crop Sciences is the property of Oil Crops Research Institute of Chinese Academy of Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective To investigate the effect and mechanism of silencing long non-coding RNA (lncRNA) LINC00472 on lipopolysaccharide (LPS)-induced renal tubular epithelial cell injury. Methods The human renal tubular epithelial cell line HK-2 (hereinafter referred to as HK-2 cells) was cultured in vitro and randomly divided into the control group, LPS group, LPS + si-LINC00472 group, LPS + si-NC group, LPS + miR-136-3p group, LPS + miR-NC group group, LPS + si-LINC00472 + anti-miR-136-3p group, and LPS + si-LINC00472 + anti-miR-NC group. Cells in the LPS + si-LINC00472 group were transfected with si-LINC00472, LPS + si-NC group with si-NC, LPS + miR-136-3p group with miR-136-3p, LPS + miR-NC group with miR-NC, LPS + si-LINC00472 + anti-miR- 136-3p group with si-LINC00472 + anti-miR-136-3p, and LPS + si-LINC00472 + anti-miR-NC group with si-LINC00472 + anti-miR-NC, followed by 1 μg/mL LPS stimulation for 24 h after 6-hour transfection, respectively. The cells in the LPS group were not transfected and were only stimulated with 1 μg/mL LPS for 24 h. The cells in the control group were routinely cultured without transfection and LPS stimulation. After the intervention, the cells in each group were collected, and RT-qPCR was used to detect the expression of lncRNA LINC00472 and miR-136-3p. The levels of IL-6 and IL-8 in the culture supernatant were detected by ELISA, and the binding sites of lncRNA LINC00472 and miR-136-3p, miR-136-3p and PDCD4 were predicted by Lnc-Base Predicted v.2 online prediction website and starBase database. The targeted regulatory relationship between lncRNA LINC00472 and miR-136-3p, miR-136-3p and PDCD4 was verified by dual-luciferase reporter gene experiments, and the protein expression of PDCD4 was detected by Western blotting. Results The relative expression of lncRNA LINC00472 in the LPS group was significantly higher than that in the control group, the relative expression of miR-136-3p was significantly lower than that in the control group, the cell viability was also significantly lower than that in the control group, and the levels of IL-6 and IL-8 in the culture supernatant were significantly higher than those of the control group (all P< 0. 05). The relative expression of lncRNA LINC00472 in the LPS + si-LINC00472 group was significantly lower than that in the LPS + si-NC group, the relative expression of miR-136-3p was significantly higher than that in the LPS + si-NC group, the cell activity was also significantly higher than that in the LPS + si-NC group, and the levels of IL-6 and IL-8 in the culture supernatant were significantly lower than those in the control group (all P<0. 05). The relative expression level of miR-136-3p and cell activity in LPS + miR-136-3p group were significantly higher than those in the LPS + miR-NC group, while the apoptosis rate, the levels of IL-6 and IL-8 in the culture supernatant and the relative expression of PDCD4 protein were significantly lower than those in the LPS + miR-NC group (all P<0. 05). The relative expression level of miR- 136-3p and the cell activity in the LPS + si-LINC00472 + anti-miR-136-3p group were significantly lower than those in LPS + si-LINC00472 + anti-miR-NC group, but the levels of IL-6 and IL-8 in culture supernatant and the relative expression of PDCD4 protein were significantly higher than those in the LPS + si-LINC00472 + anti-miR-NC group (all P<0. 05). Predicted by LncBase Predicted v.2 online prediction website and starBase database and verified by dual luciferase reporter gene experiments, lncRNA LINC00472 had a binding site with miR-136-3p, and miR-136-3p had a binding site with PDCD4. Conclusion Silencing lncRNA LINC00472 could alleviate LPS-induced renal tubular epithelial cell injury by promoting the expression of miR-136-3p and inhibiting the expression of PDCD4. [ABSTRACT FROM AUTHOR]

Copyright of Food Research & Development is the property of Food Research & Development Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background and purpose: Golgi membrane protein 1 (GOLM1) plays the role of an oncogene in lung adenocarcinoma (LUAD), however, the effects of GOLM1 on the proliferation, invasion and migration of LUAD cells and its mechanism are not clear yet. This study investigated the effects of GOLM1 on the proliferation, invasion and migration of LUAD cells and its mechanism of action. Methods: We selected cancer tissues and corresponding paracancerous tissue specimens from 90 LUAD patients who underwent surgical resection in Karamay Central Hospital from April 2019 to April 2021. The expression of GOLM1 protein in LUAD tissues and paracancerous tissues was detected by immunohistochemistry, and the relationship between GOLM1 protein expression and clinicopathological characteristics of LUAD tissues was analyzed. Western blot was used to detect the expression of GOLM1 protein in human lung epithelial cells BEAS-2B and human lung adenocarcinoma H460, A549, PG49 and H1299 cells. Lung adenocarcinoma A549 cells in logarithmic growth stage were randomly divided into blank group (cells not transfected), GOLM1 small interfering RNA negative control (si-NC) group (cells transfected with si-NC), GOLM1 small interfering RNA (si-GOLM1) group (cells transfected with si-GOLM1), insulin-like growth factor-1 (IGF-1) group [10 μmol/L phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway activator IGF-1 for 30 min] and si-GOLM1+IGF-1 group (after treatment with 10 μmol/L IGF-1 for 30 min, si-GOLM1 was transfected). Cell counting kit-8 method was used to detect cell proliferation in each group of lung adenocarcinoma A549 cells. Scratch test was used to detect cell migration in each group of lung adenocarcinoma A549 cells. Transwell experiment was used to detect cell invasion in each group of lung adenocarcinoma A549 cells. Western blot was used to detect the expressions of GOLM1 and PI3K/AKT/mTOR signaling pathway related proteins in each group of lung adenocarcinoma A549 cells. Xenograft model was constructed by subcutaneously injecting A549 cells on the right side of BALB/c nude mice, which were divided into: nude mice blank group, nude mice si-NC group, nude mice si-GOLM1 group, nude mice IGF-1 group, nude mice si-GOLM1+IGF-1 group, with 6 mice in each group. The nude mice were sacrificed six weeks after injection, the tumor was collected, and the weight and volume of the tumor were measured. Results: The results of immunohistochemistry showed that the positive expression rate of GOLM1 protein was significantly higher in LUAD tissues than in adjacent tissues (P<0.05). The expression of GOLM1 protein was significantly correlated with the degree of tumor differentiation, lymph node metastasis, and clinical stage (P<0.05), but not significantly correlated with gender, age and smoking status of LUAD patients (P>0.05). Compared with BEAS-2B cells, the relative expression level of GOLM1 protein in human lung adenocarcinoma H460, A549, PG49 and H1299 cells was significantly increased (P<0.05), and the relative expression level of GOLM1 protein in A549 cells was the highest. Therefore, A549 cells were selected for subsequent experiments. Compared with the blank group and the si-NC group, OD value, scratch healing rate, number of invaded cells, GOLM1, p-PI3K/PI3K, p-AKT/AKT and p-mTOR/mTOR protein relative expression levels in the lung adenocarcinoma A549 cells of the si-GOLM1 group were significantly reduced (P<0.05). In the lung adenocarcinoma A549 cells of the IGF-1 group, there was no significant change in GOLM1 protein, and the other corresponding indicators were significantly increased (P<0.05). Compared with the si-GOLM1 group, the OD value, scratch healing rate, number of invaded cells, p-PI3K/PI3K, p-AKT/AKT and p-mTOR/mTOR protein relative expression levels of lung adenocarcinoma A549 cells in the si-GOLM1+IGF-1 group were significantly increased (P<0.05), and there was no significant difference in GOLM1 protein relative expression level (P>0.05). Compared with the nude mice blank group and nude mice si-NC group, the mass and volume of transplanted tumors in the nude mice si-GOLM1 group were significantly reduced, while the mass and volume of transplanted tumors in the nude mice IGF-1 group were significantly increased (P<0.05). Compared with the nude mice si-GOLM1 group, the mass and volume of transplanted tumors in the nude mice si-GOLM1+IGF-1 group were significantly increased (P<0.05). Conclusion: Silencing GOLM1 gene can inhibit the activation of PI3K/AKT/mTOR signaling pathway, thereby inhibiting the proliferation, migration and invasion of lung adenocarcinoma A549 cells. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Modern Laboratory Medicine is the property of Journal of Modern Laboratory Medicine Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Applied Entomology is the property of Chinese Journal of Applied Entomology, Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective To investigate the effect of siRNA mediated KDM3A gene silencing on cell proliferation and apoptosis of breast cancer cell line MDA-MB-231. Methods Constructed a KDM3A gene-specific siRNA lentiviral vector to infect breast cancer MDA-MB-231 cells, and detected the efficiency of breast cancer cells transfected with siRNA-KDM3A by the real-time fluorescent quantitative PCR. CCK-8 method was used to detect cell proliferation, Western blot was used to test cell apoptosis and Transwell method was used to detect cell invasion. Results Compared with the control group,the expression of KDM3A mRNA was down-regulated in KDM3A-sh1, KDM3A-sh2 and KDM3A-sh3 cells, and the most efficient interference RNA was KDM3A-sh2, with the expression of mRNA decreased by 80.1%(P＜0.01).After silencing KDM3A, the proliferation of MDA-MB-231 cells decreased, the apoptosis rate increased (P＜0.05), and the invasion ability decreased (P＜0.01). Conclusions siRNA-mediated KDM3A gene silencing inhibits proliferation and invasion of breast cancer cells and promotes their apoptosis.

Copyright of Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao is the property of Chinese Journal of Preventive Veterinary Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

[ Objective] This paper aimed to overcome the easy breakdown of conventional disease resistant plants. [ Method] A new kind of transgenic rice plant5 was generated based on Host Induced Gene Silencing ( HIGS) technology. [Results] SEC11, a key subunit of signal peptide complex in Magnaporthe oryzae, was selected for generating the HIGS expression vector by gateway technology. HIGS vector was transferred into the Japonica rice cultivar Nipponbare by agrobacterium mediated transformation to obtain transgenic rice plants. The spray assay with conidia of rice blast indicated that the HIGS transgenic rice plants were improved in rice blast resistance. [Conclusion] This study lays the foundation for molecular breading of rice blast resistant varieties with HIGS technology. [ABSTRACT FROM AUTHOR]

BACKGROUND: S100A4 is a member of S100 family of calcium binding proteins, which can bind to the corresponding target proteins, and then have an important impact on the proliferation and metastasis of tumor cells and pluripotent stem cells. Recent studies have shown that ultrasound targeted destruction of microbubbles can enhance gene transfection efficiency. OBJECTIVE: To study the effect of S100A4 gene silencing by ultrasound targeted microbubble destruction on the stemness and epithelial-mesenchymal transformation of gastric cancer stem cells. METHODS: SGC-7901 human gastric cancer stem cells in logarithmic growth phase were harvested to isolate tumor stem cell spheres by serum-free suspension culture. The expression of EpCAM+CD44+ on the surface of tumor stem cells was detected by flow cytometry. The expression plasmid targeting S100A4 gene was constructed and transfected into EpCAM+CD44+ gastric cancer stem cells by ultrasound microbubble. At 48 hours after transfection, S100A4 expression was detected by western blot assay. The proliferation, clone formation, migration and invasion of gastric cancer stem cells were detected by CCK-8, clone formation test and Transwell cell. The expression of markers related to epithelial mesenchymal transformation was detected by western blot assay and qRT-PCR. RESULTS AND CONCLUSION: Flow cytometry results showed that the proportion of EpCAM+CD44+ cells after enrichment of gastric cancer stem cells was significantly higher than that before enrichment (P < 0.05). The expression of S100A4 protein in the transfected group was significantly lower than that in the untransfected group (P < 0.01). At 48 hours after transfection, the proliferation rate, clone-forming ability, migration and invasion ability of gastric cancer stem cells significantly decreased, the expression of E-cadherin increased, and the expression of N-cadherin and Vimentin decreased. These findings indicate that RNA interference with S100A4 gene can inhibit the proliferation, clone-forming ability, migration and invasion of gastric cancer stem cells by regulating the expression of E-cadherin, N-cadherin and Vimentin. [ABSTRACT FROM AUTHOR]

[Objectives] The purpose of this study was to verify the function of MDHAR2 in multigene family of monodehydroascorbate reductase (MDHAR) and to explore its effect on ascorbic acid (AA) content in non-heading Chinese cabbage. [Methods] The full length of BeMDHAR2 was amplified from cDNA of non-heading Chinese cabbage Suzhouqing. Bioinformatics was used to analyze the amino acid sequence. Sub cellular localization was analyzed by transient expression in tobacco, and real-time fluorescence quantitative PCR (RT-PCR) was used to detect the expression of the gene under different abiotic stresses and hormones treatments. The effect of the gene expression on the ascorbic acid content of Suzhouqing was verified by vinus induced gene silencing (VIGS). [Results] The gene contained an open reading frame(ORF) of 1 308 bp. encoding 435 amino acids. The phylogenetic tree analysis showed that BeMDHAR2 had the closest evolutionary relationship with Brassica rapa. BcMDHAR2 was located on the nucleus, cytoplasm and membrane. RT-qPCR showed that the expression of BeMDHARZ was up-regulated and then down-regulated under low temperature salt, heavy metal ions and exogenous hormones. The gene had a certain response to photoperiod, and the expression level increased with the light time. The expression level decreased after dark. After BMDHAR2 gene silencing, the expression level of this gene was significantly reduced, and AsA content also decreased. The expression levels of related genes in AsA-GSH cycle were also changed. The SOD activity increased in the plant. [Conclusions] The expression level of BeMDHAR2 changed with the changes of external conditions. BeMDHAR2 could be induced by different hormones and respond to different stresses, which had positive regulation on AsA content in non-heading Chinese cabbage. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Modern Laboratory Medicine is the property of Journal of Modern Laboratory Medicine Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

To compare the silencing efficiency of different virus-induced gene silencing (VIGS) systems in cotton. [Method] In this study, the kanamycin-resistant gene nptⅡ driven by the 35S promoter was used as the target gene. The RNA virus vector TRV and the DNA virus vector CLCrV were used to construct a gene silencing vector to study the gene silencing efficiency of these two sets of VIGS systems in cotton And the duration of silence. [Results] The TRV system has higher silencing efficiency than the CLCrV system, and the TRV system has a shorter silence duration than the CLCrV system. The CLCrV system can play a silent role in the reproductive organs such as cotton buds, flowers, and anthers. [Conclusion] The TRV virus silencing system is mainly suitable for short-term cotton endogenous gene silencing, and the CLCrV virus silencing system is suitable for studying the gene functions related to reproductive growth in the late stage of cotton growth and development. This study provides a reference for selecting a suitable VIGS system for cotton, which is of great significance to the study of cotton gene function. [ABSTRACT FROM AUTHOR]

To explore the effect of hypoxia-inducible factor 1α（HIF-1α）silencing on hypoxia-induced transdifferentiation in human lens epithelial cells（HLECs）. Methods HLEC-B3 cells in logarithmic growth phase were randomly divided into four groups：the normal oxygen group，the hypoxia group，negative siRNA hypoxia group，and the HIF-1α siRNA hypoxia group. HLEC-B3 in the hypoxia group，HIF-1α siRNA hypoxia group and negative siRNA hypoxia group were cultured in hypoxic culture medium（100 μmol/L CoCl2）for 24 h. HLEC-B3 cells in negative siRNA hypoxia group and HIF-1α siRNA hypoxia group were cultured under hypoxic conditions for 24 h，then transfected with negative siRNA and HIF-1α siRNA. The expression of HIF-1α，E-cadherin and α-SMA mRNA in HLEC-B3 cells were detected by qPCR. The expression of HIF-1α，E-cadherin and α-SMA protein in HLEC-B3 cells were measured with Western blot⁃ ting. Results Compared with the normal oxygen group，the relative expression of HIF-1α mRNA and protein in the hy⁃ poxia group and the negative siRNA hypoxia group increased，and the relative expression of HIF-1α mRNA and protein in the HIF-1α siRNA hypoxia group decreased（all P <0. 05）. Compared with the normal oxygen group，the relative expres⁃ sion of E-cadherin mRNA and protein in the hypoxia group and the negative siRNA hypoxia group decreased，and the rela⁃ tive expression of α-SMA mRNA and protein increased（all P<0. 05）. Compared with the hypoxia group and the negative siRNA hypoxia group，the relative expression of E-cadherin mRNA and protein in the HIF-1α siRNA hypoxia group in⁃ creased，and the relative expression of α-SMA mRNA and protein decreased（all P<0. 05）. Conclusion HIF-1α gene si⁃ lencing can inhibit the process of hypoxia-induced HLECs transdifferentiation. [ABSTRACT FROM AUTHOR]

Objective To observe the effect of silencing partitoning defective gene 3（PARD3）on the migration and invasion of cervical cancer cell line SiHa，and to explore its mechanism. Methods SiHa cells were divided into the siR⁃ NA1 group，siRNA2 group，and siRNC group. SiHa cells in the siRNA1 group were transfected with PARD3 small inter⁃ ference RNA1（siRNA1），the siRNA2 group with PARD3 small interference RNA2（siRNA2），and the siRNC group with blank vector，and then they were incubated for 48 h after transfection. Transwell migration and invasion assay was used to observe the migration and invasion abilities of the three groups，which were represented by the relative number of migration cells and the relative number of invasion cells，respectively. Pard3，JAK2，STAT3，E-cadherin and Vimentin mRNAs were detected by real-time quantitative PCR. Pard3，JAK2，STAT3，E-cadherin，Vimentin，p-JAK2，and p-STAT3 pro⁃ teins in the three groups were detected by Western blotting. Results The relative number of migration cells and the rela⁃ tive number of invasion cells in the siRNA1 and siRNA2 groups were higher than those in the siRNC group（all P<0. 05）. The relative expression levels of PARD3，E-cadherin mRNA and protein in the siRNA1 and siRNA2 groups were lower than those in the siRNC group（all P<0. 05），and the relative expression levels of Vimentin mRNA，Vimentin protein，pJAK2 protein and p-STAT3 protein in the siRNA1 and siRNA2 groups were higher than those in the siRNC group（all P< 0. 05）. Conclusion Silencing PARD3 gene can promote the migration and invasion of SiHa cells，and the mechanism may be related to the regulation of JAK2/STAT3 phosphorylation signal transduction pathway. [ABSTRACT FROM AUTHOR]

[Objectives]The paper aimed to clone BcMAX1 gene and its promoter, analyze the expression pattern of BcMAX1 and explore its function in tillering regulation. [Methods]BcMAX1 gene and its promoter were cloned, and the bioinformatics method was used to analyze the gene and promoter sequence, the protein sequence encoded by the gene, and the evolutionary relationship of MAX1 among different species. The relative expression levels of BcMAX1 in different tissues(root, stem, leaf and axillary)at different growth stages in ‘Maertou’ (the tillering variety) and ‘Suzhouqing’ (the non, tillering variety) were also compared. The function of BcMAX1 gene was preliminarily studied in Arabidopsis thaliana transgenic plants and non, heading Chinese cabbage silent plants. The expression levels of BcMAX1 and strigolactones response genes BcMAX2 and BcD14 were detected by RT, qRCR. [Results]The BcMAX1 gene contained an open reading frame of 1 593 bp, encoding 531 amino acids. MAX1 was highly conserved among different species. The promoter of BcMAX1 contained the multiple motifs that determine transcription initiation and efficiency, and the motifs that respond to drought and light. BcMAX1 gene played an important role in the process of the axillary buds elongation and tiller formation in ‘Maertou’ and ‘Suzhouqing’, and the main expression sites were root, leaf and axillary bud. BcMAX1 overexpression transgenic A. thaliana plants had fewer lateral branches, and the expression levels of BcMAX1 gene and strigolactones response genes BcMAX2 and BcD14 increased in overexpressed A. thaliana plants, but decreased in non-heading Chinese cabbage which silenced endogenous BcMAX1. [Conclusions] The expression of BcMAX1 gene in ‘Maertou’ and ‘Suzhouqing’ was spatiotemporal and tissue-specific. It was involved in the negative regulation of plant tillering and the positive regulation of the expression of strigolactones response genes BcMAX2 and BcD14. [ABSTRACT FROM AUTHOR]

[Objectives]This article aimed to explore the characteristics and regulatory functions of BcEGL3 gene in non-heading Chinese cabbage purple material and its green mutant. [Methods]In this experiment, the non-heading Chinese cabbage purple inbred line NJZX1-3 and its green mutant NJZX1-0 were used as materials to clone the BcEGL3 gene. Then BcEGL3 protein was analyzed for subcellular location by constructing the expression vector pRI101-BcEGL3, and by constructing the silencing vector pTY-BcEGL3, the changes of the anthocyanin content in the materials and the gene expression levels of BcEGL3 and BcDFR genes in NJZX1-3, NJZX1-0, pTY-S and pTY-BcEGL3 plants were analyzed. In the end, by shading NJZX1-3 materials, the accumulation of anthocyanins and the changes in the transcription level of BcEGL3 gene were analyzed. [Results]The EGL3 gene sequence cloned from NJZX1-3 and NJZX1-0 was completely identical. The sequence was 1 821 bp in length, containing an open reading frame (ORF)of 1 818 bp, encoding 606 amino acids, and named BcEGL3. The analysis of protein structure showed that EGL3 protein belonged to the bHLH-MYC-N super family, and its protein structure was relatively simple. The phylogenetic tree results showed that BcEGL3 protein had the closest relationship with BraEGL3 in Chinese cabbage, and the homology was as high as 99.9%. The results of subcellular localization showed that BcEGL3 protein was localized in the nucleus. The gene silencing results showed that, compared with NJZX1-3 plant, the leaves of transgenic pTY-BcEGL3 or pTY-S plants became lighter in purple and the corresponding anthocyanin content decreased. RT-qPCR results showed that the relative expression levels of BcEGL3 and BcDFR genes in pTY-BcEGL3 plants were lower than those in transgenic pTY-S and NJZX1-3 plants, and the color of plant leaves changed from purple to green, and the anthocyanin content significantly decreased. But the total chlorophyll content and carotenoid content did not change significantly. The anthocyanin content in non-heading Chinese cabbage leaves decreased by 68% compared with the control after shading treatment for 5 days, and the anthocyanin content in leaves decreased the most on 11 days. In addition, the BcEGL3 gene showed a downward trend after shading, with the largest decline at 14 d compared with the control. [Conclusions]BcEGL3 was located in the nucleus. BcEGL3 gene silencing could inhibit the synthesis of anthocyanins in leaves, and was closely related to the expression of BcDFR gene, and played an important role in the process of non-heading cabbage anthocyanin synthesis. [ABSTRACT FROM AUTHOR]

: [Objectives] The paper aimed to study the expression pattern of YUCCA family gene BrYUCCA5.1 in heading and non-heading Chinese cabbage and explore its function during the leaf development of Chinese cabbage. [Methods] To gain insightful information of the putative YUCCA protein, the phylogenetic relationship, conserved domain and its chemical prope1iies were conducted by bioinformatics analysis software. Quantitative real-time PCR( RT-qPCR) was performed to explore the expression level of BrYUCCA5.1 gene in different segments of heading Chinese cabbage W30, non-heading Chinese cabbage 082 and its F2 generation. Turnip yellow mosaic virus( TYMV )-derived virus induced gene silencing( VIGS) technology was employed to knock down the expression of BrYUCCA5.1 gene in non-heading Chinese cabbage 082,and study the function of this gene. [Results] The protein encoded by BrYUCCA5.1 gene of heading cabbage had high homology with Arabidopsis thaliana, Brassica oleracea, Brassica nap us and Raphanus sativus. The core regions showed high conservation with structures of FMO-like protein. Compared with other individual non-heading plants,BrYUCCA5.1was highly expressed in the middle and basal region of the leaves in heading Chinese cabbage W30,as well as the middle region of F2 generation. Using VIGS technology to silence BrYUCCA5.1 of non-heading Chinese cabbage, the silencing efficiency reached 84%. Compared with the control group, Br YUCCA5.1-pTY susceptible plants showed symptoms of virus infection on the leaves of a large area, the leaves curled and shrunk, and the :morphology was teleaxialization. [ Conclusions]BrYUCCA5.1gene pmiicipated in the regulation of leaf development of cabbage and played important role in the development of leaf bulbs of heading Chinese cabbage. [ABSTRACT FROM AUTHOR]