AIM:To investigate the expression of interleukin (IL)-22 and its related proteins in biopsy specimens from patients with ulcerative colitis (UC) and UC-related carcinogenesis. METHODS:Biopsy specimens were obtained fr...AIM:To investigate the expression of interleukin (IL)-22 and its related proteins in biopsy specimens from patients with ulcerative colitis (UC) and UC-related carcinogenesis. METHODS:Biopsy specimens were obtained from patients with inactive (n = 10), mild-to-moderately active (n = 30), severely active (n = 34), initial (n = 30), and chronic UC (n = 44), as well as UC patients with dysplasia (n = 10). Specimens from patients without colonic abnormalities (n = 20) served as controls. Chronic colitis in experimental mice was induced by 2.5% dextran sodium sulfate. The expression levels of IL-22, IL-23, IL-22R1 and phosphorylated STAT3 (p- STAT3) were determined by immunohistochemistry. Bcl-2, cyclin D1 and survivin expression was detected by Western blotting. RESULTS:Patients with active UC had significantly more IL-22, IL-23, IL-22R1 and p-STAT3-positive cells than the patients with inactive UC and normal controls. Furthermore, IL-22 and related proteins were closely related to the severity of the colitis. The expression of IL-22 and IL-22R1 in the tissue of initial UC was stronger than in that of chronic UC, whereas the expression of p-STAT3 was significantly increased in chronic UC tissues. In dysplasia tissues, the expression level of IL-22 and related proteins was higher compared with controls. Mouse colitis model showed that expression of IL-22, IL-22R1 and IL-23 was increased with time, p-STAT3 and the downstream gene were also remarkably upregulated.CONCLUSION:IL-22/STAT3 signaling pathway may be related to UC and UC-induced carcinogenesis and IL-22 can be used as a biomarker in judging the severity of UC.展开更多
Objective Along with changes in the ecology system and under the influence of various environmental factors, the incidence of tumor has been increasing year after year. There is a trend in cancer therapy to move to co...Objective Along with changes in the ecology system and under the influence of various environmental factors, the incidence of tumor has been increasing year after year. There is a trend in cancer therapy to move to combined therapies involving surgery, radiation chemotherapy and gene therapy. Cancer gene therapy in recent years has brought new opportunities for treatment of tumor. Its advantages include low rate of tolerance, insensitivity to cell cycles, high specificity and coverage for both primary and metastatic tumors1'2. However, this is a new field of clinical research. Regarding the correlation among the STAT3, CyclinD1 and P21 genes and tumors, research has focused on their expression and regulation. This article provides a summary of related research.展开更多
AIN To study the relationship of Imp2 and DR3genes with type Ⅰ diabetes mellitus.NETHODS Imp2 genotypes and DR3 wereidentified in 68 patients with type Ⅰ diabetesmellitus(Ⅰ-DM)and 71 healthy controls.Then,Ⅰ-DM pat...AIN To study the relationship of Imp2 and DR3genes with type Ⅰ diabetes mellitus.NETHODS Imp2 genotypes and DR3 wereidentified in 68 patients with type Ⅰ diabetesmellitus(Ⅰ-DM)and 71 healthy controls.Then,Ⅰ-DM patients and controls were respectivelyallocated into DR3-positive and DR3-negativegroups.The frequencies of Imp2 and DR3 genein random subjects,and Imp2 genotypes in DR3-matched subjects were compared between Ⅰ-DMpatients and controls.At the same time,Ⅰ-DMpatients were divided into 3 groups based on theonset age of diabetics:group A≤14 years,group B 15-30 years and group C≥31 years.RESULTS The frequency of DR3 in Ⅰ-DMpatients was significantly higher than that incontrols(47% vs 21%,P【0.005),and it wassignificantly higher in group A than that in groupB+C(70% vs 36%,x^2=7.07,P【0.01).Therewas a significant difference among groups withdifferent onset age of diabetics(x^2=8.19,rp=0.33,P【0.05).In random subjects,thefrequency of Imp2.R/R in Ⅰ-DM patients waslower(43% vs 61%,P【0.05)and Imp2.R/Hhigher(53% vs 28%,P【0.05)than that incontrols,and there was no significant differenceamong groups with different onset age ofdiabetics.In DR3-positive subjects,thefrequency of Imp2.R/R in Ⅰ-DM patients waslower(47% vs 87%,P【0.05)and Imp2-R/H higher(47% vs 13%,P【0.05)than that incontrols.In DR3-negative subjects,thefrequency of Imp2.R/H in Ⅰ-DM patients washigher than that in controls(58% vs 32%,P【0.01),but the frequency of Imp2-R/R and Imp2H/H was not significantly different betweenthese two groups.CONCLUSION DR3 gene may be one of thesusceptible genes of Ⅰ-DM,and significantlyrelated to the onset age of diabetics,and thepersons with DR3 may have an younger onsetage of diabeteS.The Imp2-R/R may be theprotective genotype of Ⅰ-DM,and Imp2-R/H thesusceptible genotype.These were not affectedby DR3 gene.Imp-2 genotypes were not relatedwith the onset age of diabetics.展开更多
Xanthomonas oryzea pv.oryzae(Xoo)is the causal agent of bacterial blight of rice,which is a significant threat to many of rice-growing regions.The type Ⅲ secretion system(T3SS)is an essential virulence factor in Xoo....Xanthomonas oryzea pv.oryzae(Xoo)is the causal agent of bacterial blight of rice,which is a significant threat to many of rice-growing regions.The type Ⅲ secretion system(T3SS)is an essential virulence factor in Xoo.Expression of the T3SS is often induced in the host environment or in hrp-inducing medium but is repressed in nutrient-rich medium.The elucidation of molecular mechanism underlying induction of T3SS genes expression is a very important step to lift the veil on global virulence regulation network in Xoo.Thus,an efficient and reliable genetic tool system is required for detection of the T3SS proteins.In this study,we constructed a protein expression vector pH3-flag based on the backbone of pHM1,a most widely used vector in Xoo strains,especially a model strain PXO99A.This vector contains a synthesized MCS-FLAG cassette that consists of a multiple cloning site(MCS),containing a modified pUC18 polylinker,and Flag as a C-terminal tag.The cassette is flanked by transcriptional terminators to eliminate interference of external transcription enabling detection of accurate protein expression.We evaluated the potential of this expression vector as T3SS proteins detection system and demonstrated it is applicable in the study of T3SS genes expression regulation in Xoo.This improved expression system could be very effectively used as a molecular tool in understanding some virulence genes expression and regulation in Xoo and other Xanthomonas spp.展开更多
This study examined the role of EMP-1 in tumorigenesis of non-small cell lung carcinoma (NSCLC) and the possible mechanism. Specimens were collected from 28 patients with benign lung diseases and 28 with NSCLC, and im...This study examined the role of EMP-1 in tumorigenesis of non-small cell lung carcinoma (NSCLC) and the possible mechanism. Specimens were collected from 28 patients with benign lung diseases and 28 with NSCLC, and immunohis to chemically detected to evaluate the correlation of EMP-1 expression to the clinical features of NSCLC. Recombinant adenovirus was constructed to over-express EMP-1 and then infect PC9 cells. Cell proliferation was measured by Ki67 staining. Western blotting was performed to examine the effect of EMP-1 on the PI3K/AKT signaling. Moreover, tumor xeno-grafts were established by subcutaneous injection of PC9 cell suspension (about 5×107/mL in 100 μL of PBS) into the right hind limbs of athymic nude mice. The results showed EMP-1 was significantly up-regulated in NSCLC patients as compared with those with benign lung diseases. Over-expression of EMP-1 promoted proliferation of PC9 cells, which coincided with the activation of the PI3K/AKT pathway. EMP-1 promoted the growth of xenografts of PC9 cells in athymic nude mice. It was concluded that EMP-1 expression may contribute to the development and progress of NSCLC by activating PI3K/AKT pathway.展开更多
Plant mitochondrial phosphate transporters regulate phosphate transport and ATP synthesis. Determining whether they function in abiotic stress response process would shed light on their response to salt stress. We use...Plant mitochondrial phosphate transporters regulate phosphate transport and ATP synthesis. Determining whether they function in abiotic stress response process would shed light on their response to salt stress. We used the CRISPR/Cas9 gene-editing system to mutagenize two mitochondrial phosphate transporters, OsMPT3;1 and OsMPT3;2, to investigate their regulatory roles under salt stress. Two cas9(CRISPR-associated protein9)-free homozygous mutants, mpt33 and mpt30, were confirmed to be stable. Both OsMPT3;1 and OsMPT3;2 were markedly induced by salt stress, and their mutagenesis strongly inhibited growth and development, especially under salt stress. Mutagenesis sharply reduced the accumulation of ATP, phosphate, calcium, soluble sugar, and proline and increased osmotic potential, malondialdehyde, and Na^+ /K^+ ratio under salt stress. Both mutants demonstrate normal growth and development in the presence of ATP, revealing high sensitivity to exogenous ATP under salt stress. The mutants showed lowered rates of Na^+ efflux but also of K^+ and Ca^(2+) influx under salt stress. Mutagenesis of OsMPT3;2 altered the enrichment profiles of differentially expressed genes involved mainly in synthesis of secondary metabolites, metabolism of glycolysis, pyruvate, tricarboxylic acid cycle, in response to salt stress. The mutant displayed significant accumulation differences in 14 metabolites involved in 17 metabolic pathways, and strongly up-regulated the accumulation of glutamine, a precursor in proline synthesis, under salt stress. These findings suggest that the OsMPT3 gene modulates phosphate transport and energy supply for ATP synthesis and triggers changes in accumulation of ions and metabolites participating in osmotic regulation in rice under salt stress, thus increasing rice salt tolerance. This study demonstrates the effective application of CRISPR/Cas9 gene-editing to the investigation of plant functional genes.展开更多
Summary: A new type of TGF-β3 fusion protein with targeted therapy function was constructed, and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 g...Summary: A new type of TGF-β3 fusion protein with targeted therapy function was constructed, and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 gene into adipose-derived stem cells (ADSCs). The recombinant pIRES- EGFP-MMP was constructed by inserting the sense and antisense DNA of encoding the amino acid of the synthetic MMP enzyme cutting site into the eukaryotic expression vector pIRES-EGFE LAP and mTGF-β3 fragments were obtained by using RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, and the recombinant plasmid of pIRES-EGFP- LAP-MMP-mTGF-β3 was constructed, which was transferred to ADSCs. The ADSCs were cultured and divided in three groups: experimental group (MMP group), negative control group (no MMP) and non-transfection group. The morphological changes were observed microscopically, and the expression of proteoglycan and type II collagen (Col II) was detected by using Alcian blue staining and immuno- histochemistry staining at 7th, 14th and 21st day after culture. The recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3 was correctly constructed by methods of enzyme cutting and se- quencing analysis. The mTGF-β3 fusion protein was successfully expressed after transfection, and in the presence of the MMP, active protein mTGF-β3 was generated, which significantly promoted differ- entiation of ADSCs into chondrocytes and the expression of cartilage matrix. The novel fusion protein LAP-MMP-mTGF-β3 can targetedly induce differentiation of ADSCs into chondrocytes, which would open up prospects for target therapy of cartilage damage repair in future.展开更多
Objective:To study the correlation of Runt-related transcription factor gene 3 (RUNX3) expression in osteosarcoma tissue with cell proliferation and angiogenesis.Methods: A total of 80 patients with osteosarcoma who w...Objective:To study the correlation of Runt-related transcription factor gene 3 (RUNX3) expression in osteosarcoma tissue with cell proliferation and angiogenesis.Methods: A total of 80 patients with osteosarcoma who were treated in our hospital between February 2014 and February 2017 were collected, and the RUNX3 expression in osteosarcoma tissue and adjacent tissue were detected. According to the RUNX3 expression in tumor tissue, the patients were further divided into high RUNX3 expression group and low RUNX3 expression group, and the proliferation gene and angiogenesis gene expression were compared.Results:RUNX3, KISS-1 and RanBP9 mRNA expression in osteosarcoma tissue were significantly lower than those in adjacent tissue while VCP, Six1, S100A6, IF-1α, MMP-14, bFGF and Ang-2 mRNA expression were significantly higher than those in adjacent tissue;KISS-1 and RanBP9 mRNA expression in osteosarcoma tissue of high RUNX3 expression group were significantly higher than those of low RUNX3 expression group while VCP, Six1, S100A6, IF-1 , MMP-14, bFGF and Ang-2 mRNA expression were significantly lower than those of low RUNX3 expression group.Conclusions:The desease of RUNX3 expression in osteosarcoma tissue is one of the direct causes of increased tumor proliferation activity and strong angiogenesis.展开更多
Common wheat(Triticum aestivum L.)is the most important crop in the world and a typical allopolyploid with a large and complex genome.Pre-harvest sprouting(PHS)leads to a significant reduction in grain quality worldwi...Common wheat(Triticum aestivum L.)is the most important crop in the world and a typical allopolyploid with a large and complex genome.Pre-harvest sprouting(PHS)leads to a significant reduction in grain quality worldwide.PHS is a complex trait with related QTL located on different chromosomes.However,the study of markers and genes related to PHS resistance is limited especially for whitegrained wheat.Four pairs of near isogenic lines(NILs)from a white-grained wheat cross of CharaDM5637B*8 targeting a major QTL for PHS resistance(Qphs.ccsu-3A.1)on wheat chromosme 3AL were genotyped using the 90K SNP Illumina iSelect array.Ten SNPs were identified,with a 75%-100%consistency between genotype and phenotype in the resistant or susceptible isolines.The 10 SNPs were converted to cost-effective kompetitive allele-specific PCR(KASP)markers.Screening of 48 wheat cultivars with different phenotypes of PHS identified four KASP markers with 81.3%-85.4%conformity between genotype and phenotype.Further investigation revealed that the four SNPs(BS00022245_51,Kukri_c49927_151,BS00022884_51 and BS00110550_51)corresponding to the four validated KASP markers are residing in three independent genes(TraesCS3A03G1072800,TraesCS3A03G1072400,TraesCS3A03G1071800)close to each other with a distance of 4.28-4.48 Mb to the targeted QTL.These three annotated genes have potential functions related to PHS resistance.Our study revealed that combined use of NILs and the 90K SNP chip is a powerful approach for developing KASP markers and mining functional genes in wheat.The KASP markers for PHS resistance on chromosome 3AL are useful for high-throughput evaluation and marker-assisted selection,and the three identified genes could lead to a better understanding of the genetic pathways controlling PHS.展开更多
BACKGROUND:With the objective of developing a locally- produced radioactive stent,the present study used in vivo animal experiments to explore apoptosis of proliferative smooth muscle cells resulting from facilitation...BACKGROUND:With the objective of developing a locally- produced radioactive stent,the present study used in vivo animal experiments to explore apoptosis of proliferative smooth muscle cells resulting from facilitation of the expression of genes caused byγ-radiation in order to prevent bile duct restenosis.We therefore explored the effects and significance ofγ-radiation on the activity of caspase-3,Fas and Bcl-2 genes in apoptosis of proliferative smooth muscle cells in the bile duct walls of dogs. METHODS:Twelve dogs were randomly divided into 2 groups(6 in each group).A postinjury bile duct stenosis model was established and radioactive 103 Pd( 103 palladium) or ordinary bile duct stents were implanted into the bile ducts.HE staining,RT-PCR and immunohistochemistry were used to detect the proliferation and apoptosis of bile duct smooth muscle cells in proliferative endomembrane and the expression of related caspase-3,Bcl-2 and Fas genes. RESULTS:The expression of caspase-3 and Fas genes in the bile duct tissues of dogs with radioactive stents was higher than that of dogs with ordinary stents.There was significant apoptosis of proliferative smooth muscle cells in the bile ducts.The expression of the Bcl-2 gene in the bile duct tissues of dogs with radioactive stents was lower than that in those with ordinary stents.There was significant apoptosis of proliferative smooth muscle cells in the dogs with low Bcl-2 gene expression. CONCLUSIONS:Radiation increases the activity of caspase-3 and Fas genes and is associated with apoptosis. The radioactive 103 Pd stent may facilitate apoptosis of proliferative smooth muscle cells in the bile ducts of dogs by activating these genes.The Bcl-2 gene expression level is correlated with the occurrence of apoptosis and the radiosusceptibility of cells.展开更多
The objective of this study was to determine the genotype effects of the bovine insulin-like growth factor Ⅰ (IGF-Ⅰ) and its binding protein 3 (IGFBP3) genes on growth and development traits in beef cows, includ...The objective of this study was to determine the genotype effects of the bovine insulin-like growth factor Ⅰ (IGF-Ⅰ) and its binding protein 3 (IGFBP3) genes on growth and development traits in beef cows, including 130 Chinese Simmental, 42 Nanyang, and 47 Luxi Yellow cattle. Sequence variations in the bovine IGF-Ⅰ and IGFBP3 genes were investigated by single strand conformation polymorphism (SSCP). SSCPs were detected in 6 fragments, which is the 5'-flanking region, the 2nd exon, the 5th exon, and the 5th intron of the IGF-1 gene, and the 2nd exon, the 3rd exon of the 1GFBP3 gene. Two polymorphisms, an A-to-G transition in the 2rid exon of the IGF-Ⅰ gene and a T-to-C transition in the 2rid exon of IGFBP3 gene were detected in 3 breeds. The allele frequencies of 2 polymorphisms were 0.0411 (A), 0.9589 (B), and 0.7237 (A), 0.2763 (B), respectively. These 2 loci were analyzed to associate with body weight, height at withers, body length, heart girth, rump width, and beef production index (BPI) at 0, 6, 12, 24, and 36-month old. The IGFBP3 locus was shown to be associated with rump width, heart girth at 24-month and 36-month. Animals with BB genotype had higher rump width (24.86 ± 0.47) cm at 24-month and (27.50 ± 0.63) em at 36-month. The heart girth was highest for the individuals with BB genotype (171.33 ± 1.84) cm and higher than those with AB genotype (166.68 ± 1.13) cm (P〈 0.05) at 36-month.展开更多
Imprinted genes play significant roles in the regulation of fetal growth, development, function of the placenta and postnatal behavior in mammals, but little is known in pigs. In order to investigate the imprinting st...Imprinted genes play significant roles in the regulation of fetal growth, development, function of the placenta and postnatal behavior in mammals, but little is known in pigs. In order to investigate the imprinting status of porcine retro-transposon like 1 (RTL1) and type 3 iodothyronine deiodinase (DIO3) genes, DNA or RNA samples of the parents and F1 animals, generated with reciprocal crosses between Large White and Meishan breeds, were isolated, and analyzed by reverse transcription polymerase chain reaction restriction fragment length polymorphism (RT-PCR-RFLP). The results demonstrated that the RTL1 gene was paternally expressed in 10 tissues, such as the skeletal muscle, heart, spleen, liver, kidney, lung, stomach, fat, small intestine and brain, and D103 gene exhibited paternal expression in the skeletal muscle, heart, spleen, lung, stomach, and brain, in 2-month-old pigs. The association of RTL1 and DI03 with carcass traits was further analyzed in the F2 population of Large White×Meishan pigs. The statistical results showed that the R TL1 A1101G polymorphism (EU781029) was significantly associated with lean meat percentage (LMP) and fat meat percentage (FMP) (P〈0.05), while the D103 A744C polymorphism (AY533208) was not significantly associated with any carcass traits. These results indicate that the imprinting status of RTL1 and DIO3 is well kept across the mammalian species, and porcine RTL1 may have important roles in muscle growth and fat deposition.展开更多
Background: RING H2 finger E3 ligase (RH2FE3) genes encode cysteine rich proteins that mediate E3 ubiquitin ligase activity and degrade target substrates. The roles of these genes in plant responses to phytohormone...Background: RING H2 finger E3 ligase (RH2FE3) genes encode cysteine rich proteins that mediate E3 ubiquitin ligase activity and degrade target substrates. The roles of these genes in plant responses to phytohormones and abiotic stresses are well documented in various species, but their roles in cotton fiber development are poorly understood. To date, genome wide identification and expression analyses of Gossypium hirsutum RH2FE3 genes have not been reported. Methods: We performed computational identification, structural and phylogenetic analyses, chromosomal distribution analysis and estimated KJKs values of G hirsutum RH2FE3 genes. Orthologous and paralogous gene pairs were identified by all versus all BLASTP searches. We predicted cis regulatory elements and analyzed microarray data sets to generate heatmaps at different development stages. Tissue specific expression in cotton fiber, and hormonal and abiotic stress responses were determined by quantitative real time polymerase chain reaction (qRT PCR) analysis. Results: We investigated 140 G hirsutum, 80 G. orboreum, and evolutionary mechanisms and compared them with orthologs 89 G. roimondii putative RH2FB genes and their in Arobidopsis and rice. A domain based analysis of the G hirsutum RH2FE3 genes predicted conserved signature motifs and gene structures. Chromosomal localization showed the genes were distributed across all G hirsutum chromosomes, and 60 duplication events (4 tandem and 56 segmental duplications) and 98 orthologs were detected, cis elements were detected in the promoter regions of G hirsutum RH2FE3 genes. Microarray data and qRT PCR analyses showed that G hirsutum RH2FE3 genes were strongly correlated with cotton fiber development. Additionally, almost all the (brassinolide, gibberellic acid (GA), indole 3-acetic acid drought, and salt). dentified genes were up regulated in response to phytohormones (IAA), and salicylic acid (SA)) and abiotic stresses (cold, heat, Conclusions: The genome wide identification, comprehensive analysis, and characterization of conserved domains and gene structures, as well as phylogenetic analysis, cis element prediction, and expression profile analysis of G hirsutum RH2FE3 genes and their roles in cotton fiber development and responses to plant hormones and abiotic stresses are reported here for the first time. Our findings will contribute to the genome wide analysis of putative RH2FE3 genes in other species and lay a foundation for future physiological and functional research on G hirsutum RH2FE3 genes.展开更多
Objective: To investigate whether deletion of chromosome 3 is involved in the carcinogenesis of primary glioblastoma multiforme (GBM) and to localize the possible common deletion region in the aforementioned chromosom...Objective: To investigate whether deletion of chromosome 3 is involved in the carcinogenesis of primary glioblastoma multiforme (GBM) and to localize the possible common deletion region in the aforementioned chromosome. Methods: PCR based microsatellite polymorphism analyses were performed to detect loss of heterozygosity (LOH). Twenty-three loci on chromosome 3 were examined in 20 cases of GBM. Fluorescence-labeled primers and Perkin Elmer 377 DNA Sequencer were applied. Results: 50% informative cases of GBM displayed LOH on chromosome 3. 50% of informative cases displayed LOH on 3q and 35% on 3p. 25.6% of informative loci showed LOH in our series, in which frequent LOH were observed in the chromosomal region from loci D3S1614 (42.9%) to D3S1565 (35.3%) on 3q24–27 and at loci D3S1569 (35.3%) on 3q22–23 and D3S1289 (33.3%) on 3p14.1–14.3. Conclusion: Loss of genetic material on chromosome 3 may play an important part in the tumorigenesis of GBM. The chromosomal regions from loci D3S1614 to D3S1565 on 3q24–27 and at loci D3S1569 on 3q22–23 and D3S1289 on 3p14.1–14.3 are potential sites for novel tumor suppressor genes associated with GBM.展开更多
Vegetative Insecticidal Proteins(VIPs), a large family of insecticidal proteins, are produced from Bacillus thuringiensis(Bt) during the vegetative growth stage. VIPs represent the second generation of bio-insecticide...Vegetative Insecticidal Proteins(VIPs), a large family of insecticidal proteins, are produced from Bacillus thuringiensis(Bt) during the vegetative growth stage. VIPs represent the second generation of bio-insecticides that confer a wider insecticidal spectrum and have stronger activity. This work compared the geographical distribution of Bt strains and their vip3 genes in different climatic zones in China, the tropical(Hainan Province), subtropical(Guangxi Province) and temperate zones(Heilongjiang Province). A total of 156 Bt strains were isolated from 841 soil samples in Hainan Province tropical region, 356 Bt strains from 1 420 soil samples in Guangxi Province and 167 Bt strains from 1 010 soil samples in different geographical regions in Heilongjiang Province. Twenty-two out of 156 strains from tropical Hainan Province and two out of 356 from subtropical Guangxi Province were found to express vip3 genes, while vip3 genes were not expressed from temperate zone in Heilongjiang Province. Restriction Fragment Length Polymorphism(RFLP) was used to identify different types of vip3 genes that were within the same family and three fulllength vip3 genes were isolated. The genes cloned from Bacillus thuringiensis strain SL3 expressed in the transformed E. coli BL21 strain. Through SDS-PAGE, 88.6 ku insecticidal protein was expressed. The bioassays used two-instar larva of Lepidoptera insects(Spodoptera exigua and Agrotis ipsilon) were performed. The results of the bioassays showed that the protein strongly inhibited the body weight increasement on Spodoptera exigua and Agrotis ipsilon in a standard bioassay. Taken together, the results indicated that the distribution of Bt strains and vip3 genes had regional preference. Tropical and subtropical regions were the rich resources of Bt strains and vip3 genes compared with temperate region. These results would undoubtedly facilitate the studies of insecticidal proteins and expand the list of the pest-killing candidates to make fully use of the extremely rich microbial resources. The new vip3 genes isolated in the current study might also help resolve the emerging insecticidal resistance problems.展开更多
Objective:To investigate the core target genes of miR-29b-3p,and analyze the clinical significance of the core target genes in glioma.Methods:Bioinformatics analysis was used to predict and screen the target genes of ...Objective:To investigate the core target genes of miR-29b-3p,and analyze the clinical significance of the core target genes in glioma.Methods:Bioinformatics analysis was used to predict and screen the target genes of miR-29b-3p.STRING and Cytoscape software were used to analyze the protein-protein interaction(PPI)of target genes.the differences expression and survival prognosis in glioma were analyzed by GEPIA and CGGA.Independent prognostic factors analyzed by univariate and multivariate Cox proportional hazards regression model.Results:22 target genes of miR-29b-3p were predicted using LinkedOmics,miRDB,miRTarBase,TargetScan,and starbase databases.Through the construction of the PPI network,genes out of the network were removed,and a total of 16 genes were screened for further study of their clinical significance.Based on analysis of GEPIA and CGGA databases,COL2A1,DNMT3A,and DNMT3B were excluded.Through further analysis of the univariate and multivariate Cox proportional hazard regression model,finally identified three core target genes:SERPINH1,LOXL2,CDK6.Conclusion:Bioinformatics analysis showed that miR-29b-3p targeted three core genes such as SERPINH1,LOXL2,and CDK6 in glioma.The expression of these genes was different between brain normal tissues and gliomas,between different grades of tumor,IDH mutation status and 1p/19q codeletion status.Its high expression had adverse effects on overall survival and recurrence-free survival.These core target genes can be used as an independent prognostic factor.展开更多
基金Supported by National Natural Science Foundation of China,No.81072692Natural Science Foundation of Jiangsu Higher Education Institutions of China,No.10KJB320007
文摘AIM:To investigate the expression of interleukin (IL)-22 and its related proteins in biopsy specimens from patients with ulcerative colitis (UC) and UC-related carcinogenesis. METHODS:Biopsy specimens were obtained from patients with inactive (n = 10), mild-to-moderately active (n = 30), severely active (n = 34), initial (n = 30), and chronic UC (n = 44), as well as UC patients with dysplasia (n = 10). Specimens from patients without colonic abnormalities (n = 20) served as controls. Chronic colitis in experimental mice was induced by 2.5% dextran sodium sulfate. The expression levels of IL-22, IL-23, IL-22R1 and phosphorylated STAT3 (p- STAT3) were determined by immunohistochemistry. Bcl-2, cyclin D1 and survivin expression was detected by Western blotting. RESULTS:Patients with active UC had significantly more IL-22, IL-23, IL-22R1 and p-STAT3-positive cells than the patients with inactive UC and normal controls. Furthermore, IL-22 and related proteins were closely related to the severity of the colitis. The expression of IL-22 and IL-22R1 in the tissue of initial UC was stronger than in that of chronic UC, whereas the expression of p-STAT3 was significantly increased in chronic UC tissues. In dysplasia tissues, the expression level of IL-22 and related proteins was higher compared with controls. Mouse colitis model showed that expression of IL-22, IL-22R1 and IL-23 was increased with time, p-STAT3 and the downstream gene were also remarkably upregulated.CONCLUSION:IL-22/STAT3 signaling pathway may be related to UC and UC-induced carcinogenesis and IL-22 can be used as a biomarker in judging the severity of UC.
文摘Objective Along with changes in the ecology system and under the influence of various environmental factors, the incidence of tumor has been increasing year after year. There is a trend in cancer therapy to move to combined therapies involving surgery, radiation chemotherapy and gene therapy. Cancer gene therapy in recent years has brought new opportunities for treatment of tumor. Its advantages include low rate of tolerance, insensitivity to cell cycles, high specificity and coverage for both primary and metastatic tumors1'2. However, this is a new field of clinical research. Regarding the correlation among the STAT3, CyclinD1 and P21 genes and tumors, research has focused on their expression and regulation. This article provides a summary of related research.
文摘AIN To study the relationship of Imp2 and DR3genes with type Ⅰ diabetes mellitus.NETHODS Imp2 genotypes and DR3 wereidentified in 68 patients with type Ⅰ diabetesmellitus(Ⅰ-DM)and 71 healthy controls.Then,Ⅰ-DM patients and controls were respectivelyallocated into DR3-positive and DR3-negativegroups.The frequencies of Imp2 and DR3 genein random subjects,and Imp2 genotypes in DR3-matched subjects were compared between Ⅰ-DMpatients and controls.At the same time,Ⅰ-DMpatients were divided into 3 groups based on theonset age of diabetics:group A≤14 years,group B 15-30 years and group C≥31 years.RESULTS The frequency of DR3 in Ⅰ-DMpatients was significantly higher than that incontrols(47% vs 21%,P【0.005),and it wassignificantly higher in group A than that in groupB+C(70% vs 36%,x^2=7.07,P【0.01).Therewas a significant difference among groups withdifferent onset age of diabetics(x^2=8.19,rp=0.33,P【0.05).In random subjects,thefrequency of Imp2.R/R in Ⅰ-DM patients waslower(43% vs 61%,P【0.05)and Imp2.R/Hhigher(53% vs 28%,P【0.05)than that incontrols,and there was no significant differenceamong groups with different onset age ofdiabetics.In DR3-positive subjects,thefrequency of Imp2.R/R in Ⅰ-DM patients waslower(47% vs 87%,P【0.05)and Imp2-R/H higher(47% vs 13%,P【0.05)than that incontrols.In DR3-negative subjects,thefrequency of Imp2.R/H in Ⅰ-DM patients washigher than that in controls(58% vs 32%,P【0.01),but the frequency of Imp2-R/R and Imp2H/H was not significantly different betweenthese two groups.CONCLUSION DR3 gene may be one of thesusceptible genes of Ⅰ-DM,and significantlyrelated to the onset age of diabetics,and thepersons with DR3 may have an younger onsetage of diabeteS.The Imp2-R/R may be theprotective genotype of Ⅰ-DM,and Imp2-R/H thesusceptible genotype.These were not affectedby DR3 gene.Imp-2 genotypes were not relatedwith the onset age of diabetics.
基金supported by the National Key R&D Program of China (2017YFD0200400)the National Natural Science Foundation of China (31772122 and 31470235)
文摘Xanthomonas oryzea pv.oryzae(Xoo)is the causal agent of bacterial blight of rice,which is a significant threat to many of rice-growing regions.The type Ⅲ secretion system(T3SS)is an essential virulence factor in Xoo.Expression of the T3SS is often induced in the host environment or in hrp-inducing medium but is repressed in nutrient-rich medium.The elucidation of molecular mechanism underlying induction of T3SS genes expression is a very important step to lift the veil on global virulence regulation network in Xoo.Thus,an efficient and reliable genetic tool system is required for detection of the T3SS proteins.In this study,we constructed a protein expression vector pH3-flag based on the backbone of pHM1,a most widely used vector in Xoo strains,especially a model strain PXO99A.This vector contains a synthesized MCS-FLAG cassette that consists of a multiple cloning site(MCS),containing a modified pUC18 polylinker,and Flag as a C-terminal tag.The cassette is flanked by transcriptional terminators to eliminate interference of external transcription enabling detection of accurate protein expression.We evaluated the potential of this expression vector as T3SS proteins detection system and demonstrated it is applicable in the study of T3SS genes expression regulation in Xoo.This improved expression system could be very effectively used as a molecular tool in understanding some virulence genes expression and regulation in Xoo and other Xanthomonas spp.
基金supported by grants from the National Natural Science Foundation of China (Nos.81072431,30872472,30973496 and 30800569)the Innovative Foundation of Huazhong University of Science and Technology (No.2010MS027)+1 种基金the Foundation of "973" Program (No.2009CB521802)by Special Fund for Central University Basic Scientific Research (Nos.2011JC062,2011JC063)
文摘This study examined the role of EMP-1 in tumorigenesis of non-small cell lung carcinoma (NSCLC) and the possible mechanism. Specimens were collected from 28 patients with benign lung diseases and 28 with NSCLC, and immunohis to chemically detected to evaluate the correlation of EMP-1 expression to the clinical features of NSCLC. Recombinant adenovirus was constructed to over-express EMP-1 and then infect PC9 cells. Cell proliferation was measured by Ki67 staining. Western blotting was performed to examine the effect of EMP-1 on the PI3K/AKT signaling. Moreover, tumor xeno-grafts were established by subcutaneous injection of PC9 cell suspension (about 5×107/mL in 100 μL of PBS) into the right hind limbs of athymic nude mice. The results showed EMP-1 was significantly up-regulated in NSCLC patients as compared with those with benign lung diseases. Over-expression of EMP-1 promoted proliferation of PC9 cells, which coincided with the activation of the PI3K/AKT pathway. EMP-1 promoted the growth of xenografts of PC9 cells in athymic nude mice. It was concluded that EMP-1 expression may contribute to the development and progress of NSCLC by activating PI3K/AKT pathway.
基金supported by the National Key Research and Development Program of China(2016YFC0501203)the National Genetically Modified Organism Project(2016ZX08010005-9)。
文摘Plant mitochondrial phosphate transporters regulate phosphate transport and ATP synthesis. Determining whether they function in abiotic stress response process would shed light on their response to salt stress. We used the CRISPR/Cas9 gene-editing system to mutagenize two mitochondrial phosphate transporters, OsMPT3;1 and OsMPT3;2, to investigate their regulatory roles under salt stress. Two cas9(CRISPR-associated protein9)-free homozygous mutants, mpt33 and mpt30, were confirmed to be stable. Both OsMPT3;1 and OsMPT3;2 were markedly induced by salt stress, and their mutagenesis strongly inhibited growth and development, especially under salt stress. Mutagenesis sharply reduced the accumulation of ATP, phosphate, calcium, soluble sugar, and proline and increased osmotic potential, malondialdehyde, and Na^+ /K^+ ratio under salt stress. Both mutants demonstrate normal growth and development in the presence of ATP, revealing high sensitivity to exogenous ATP under salt stress. The mutants showed lowered rates of Na^+ efflux but also of K^+ and Ca^(2+) influx under salt stress. Mutagenesis of OsMPT3;2 altered the enrichment profiles of differentially expressed genes involved mainly in synthesis of secondary metabolites, metabolism of glycolysis, pyruvate, tricarboxylic acid cycle, in response to salt stress. The mutant displayed significant accumulation differences in 14 metabolites involved in 17 metabolic pathways, and strongly up-regulated the accumulation of glutamine, a precursor in proline synthesis, under salt stress. These findings suggest that the OsMPT3 gene modulates phosphate transport and energy supply for ATP synthesis and triggers changes in accumulation of ions and metabolites participating in osmotic regulation in rice under salt stress, thus increasing rice salt tolerance. This study demonstrates the effective application of CRISPR/Cas9 gene-editing to the investigation of plant functional genes.
基金supported by the National Natural Science Foundation of China(No.81101376)
文摘Summary: A new type of TGF-β3 fusion protein with targeted therapy function was constructed, and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 gene into adipose-derived stem cells (ADSCs). The recombinant pIRES- EGFP-MMP was constructed by inserting the sense and antisense DNA of encoding the amino acid of the synthetic MMP enzyme cutting site into the eukaryotic expression vector pIRES-EGFE LAP and mTGF-β3 fragments were obtained by using RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, and the recombinant plasmid of pIRES-EGFP- LAP-MMP-mTGF-β3 was constructed, which was transferred to ADSCs. The ADSCs were cultured and divided in three groups: experimental group (MMP group), negative control group (no MMP) and non-transfection group. The morphological changes were observed microscopically, and the expression of proteoglycan and type II collagen (Col II) was detected by using Alcian blue staining and immuno- histochemistry staining at 7th, 14th and 21st day after culture. The recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3 was correctly constructed by methods of enzyme cutting and se- quencing analysis. The mTGF-β3 fusion protein was successfully expressed after transfection, and in the presence of the MMP, active protein mTGF-β3 was generated, which significantly promoted differ- entiation of ADSCs into chondrocytes and the expression of cartilage matrix. The novel fusion protein LAP-MMP-mTGF-β3 can targetedly induce differentiation of ADSCs into chondrocytes, which would open up prospects for target therapy of cartilage damage repair in future.
文摘Objective:To study the correlation of Runt-related transcription factor gene 3 (RUNX3) expression in osteosarcoma tissue with cell proliferation and angiogenesis.Methods: A total of 80 patients with osteosarcoma who were treated in our hospital between February 2014 and February 2017 were collected, and the RUNX3 expression in osteosarcoma tissue and adjacent tissue were detected. According to the RUNX3 expression in tumor tissue, the patients were further divided into high RUNX3 expression group and low RUNX3 expression group, and the proliferation gene and angiogenesis gene expression were compared.Results:RUNX3, KISS-1 and RanBP9 mRNA expression in osteosarcoma tissue were significantly lower than those in adjacent tissue while VCP, Six1, S100A6, IF-1α, MMP-14, bFGF and Ang-2 mRNA expression were significantly higher than those in adjacent tissue;KISS-1 and RanBP9 mRNA expression in osteosarcoma tissue of high RUNX3 expression group were significantly higher than those of low RUNX3 expression group while VCP, Six1, S100A6, IF-1 , MMP-14, bFGF and Ang-2 mRNA expression were significantly lower than those of low RUNX3 expression group.Conclusions:The desease of RUNX3 expression in osteosarcoma tissue is one of the direct causes of increased tumor proliferation activity and strong angiogenesis.
基金funded by Global Innovation Linkage program (GIL53853) from Australian Department of Industry, Science, Energy and ResourcesAustralian Government RTP Scholarship (International)University Postgraduate Awards (UPA)
文摘Common wheat(Triticum aestivum L.)is the most important crop in the world and a typical allopolyploid with a large and complex genome.Pre-harvest sprouting(PHS)leads to a significant reduction in grain quality worldwide.PHS is a complex trait with related QTL located on different chromosomes.However,the study of markers and genes related to PHS resistance is limited especially for whitegrained wheat.Four pairs of near isogenic lines(NILs)from a white-grained wheat cross of CharaDM5637B*8 targeting a major QTL for PHS resistance(Qphs.ccsu-3A.1)on wheat chromosme 3AL were genotyped using the 90K SNP Illumina iSelect array.Ten SNPs were identified,with a 75%-100%consistency between genotype and phenotype in the resistant or susceptible isolines.The 10 SNPs were converted to cost-effective kompetitive allele-specific PCR(KASP)markers.Screening of 48 wheat cultivars with different phenotypes of PHS identified four KASP markers with 81.3%-85.4%conformity between genotype and phenotype.Further investigation revealed that the four SNPs(BS00022245_51,Kukri_c49927_151,BS00022884_51 and BS00110550_51)corresponding to the four validated KASP markers are residing in three independent genes(TraesCS3A03G1072800,TraesCS3A03G1072400,TraesCS3A03G1071800)close to each other with a distance of 4.28-4.48 Mb to the targeted QTL.These three annotated genes have potential functions related to PHS resistance.Our study revealed that combined use of NILs and the 90K SNP chip is a powerful approach for developing KASP markers and mining functional genes in wheat.The KASP markers for PHS resistance on chromosome 3AL are useful for high-throughput evaluation and marker-assisted selection,and the three identified genes could lead to a better understanding of the genetic pathways controlling PHS.
文摘BACKGROUND:With the objective of developing a locally- produced radioactive stent,the present study used in vivo animal experiments to explore apoptosis of proliferative smooth muscle cells resulting from facilitation of the expression of genes caused byγ-radiation in order to prevent bile duct restenosis.We therefore explored the effects and significance ofγ-radiation on the activity of caspase-3,Fas and Bcl-2 genes in apoptosis of proliferative smooth muscle cells in the bile duct walls of dogs. METHODS:Twelve dogs were randomly divided into 2 groups(6 in each group).A postinjury bile duct stenosis model was established and radioactive 103 Pd( 103 palladium) or ordinary bile duct stents were implanted into the bile ducts.HE staining,RT-PCR and immunohistochemistry were used to detect the proliferation and apoptosis of bile duct smooth muscle cells in proliferative endomembrane and the expression of related caspase-3,Bcl-2 and Fas genes. RESULTS:The expression of caspase-3 and Fas genes in the bile duct tissues of dogs with radioactive stents was higher than that of dogs with ordinary stents.There was significant apoptosis of proliferative smooth muscle cells in the bile ducts.The expression of the Bcl-2 gene in the bile duct tissues of dogs with radioactive stents was lower than that in those with ordinary stents.There was significant apoptosis of proliferative smooth muscle cells in the dogs with low Bcl-2 gene expression. CONCLUSIONS:Radiation increases the activity of caspase-3 and Fas genes and is associated with apoptosis. The radioactive 103 Pd stent may facilitate apoptosis of proliferative smooth muscle cells in the bile ducts of dogs by activating these genes.The Bcl-2 gene expression level is correlated with the occurrence of apoptosis and the radiosusceptibility of cells.
基金funded by the National Key Technology R&D Program of the China (2006BAD01A10,2006BAD14B07, 2006BAD04A16)the National High Technology Research and Development Program of China (2006AA10Z197)
文摘The objective of this study was to determine the genotype effects of the bovine insulin-like growth factor Ⅰ (IGF-Ⅰ) and its binding protein 3 (IGFBP3) genes on growth and development traits in beef cows, including 130 Chinese Simmental, 42 Nanyang, and 47 Luxi Yellow cattle. Sequence variations in the bovine IGF-Ⅰ and IGFBP3 genes were investigated by single strand conformation polymorphism (SSCP). SSCPs were detected in 6 fragments, which is the 5'-flanking region, the 2nd exon, the 5th exon, and the 5th intron of the IGF-1 gene, and the 2nd exon, the 3rd exon of the 1GFBP3 gene. Two polymorphisms, an A-to-G transition in the 2rid exon of the IGF-Ⅰ gene and a T-to-C transition in the 2rid exon of IGFBP3 gene were detected in 3 breeds. The allele frequencies of 2 polymorphisms were 0.0411 (A), 0.9589 (B), and 0.7237 (A), 0.2763 (B), respectively. These 2 loci were analyzed to associate with body weight, height at withers, body length, heart girth, rump width, and beef production index (BPI) at 0, 6, 12, 24, and 36-month old. The IGFBP3 locus was shown to be associated with rump width, heart girth at 24-month and 36-month. Animals with BB genotype had higher rump width (24.86 ± 0.47) cm at 24-month and (27.50 ± 0.63) em at 36-month. The heart girth was highest for the individuals with BB genotype (171.33 ± 1.84) cm and higher than those with AB genotype (166.68 ± 1.13) cm (P〈 0.05) at 36-month.
基金supported by the National Natural Sci-ence Foundation of China (30571331)the China Postdoctoral Science Foundation (2005038161)
文摘Imprinted genes play significant roles in the regulation of fetal growth, development, function of the placenta and postnatal behavior in mammals, but little is known in pigs. In order to investigate the imprinting status of porcine retro-transposon like 1 (RTL1) and type 3 iodothyronine deiodinase (DIO3) genes, DNA or RNA samples of the parents and F1 animals, generated with reciprocal crosses between Large White and Meishan breeds, were isolated, and analyzed by reverse transcription polymerase chain reaction restriction fragment length polymorphism (RT-PCR-RFLP). The results demonstrated that the RTL1 gene was paternally expressed in 10 tissues, such as the skeletal muscle, heart, spleen, liver, kidney, lung, stomach, fat, small intestine and brain, and D103 gene exhibited paternal expression in the skeletal muscle, heart, spleen, lung, stomach, and brain, in 2-month-old pigs. The association of RTL1 and DI03 with carcass traits was further analyzed in the F2 population of Large White×Meishan pigs. The statistical results showed that the R TL1 A1101G polymorphism (EU781029) was significantly associated with lean meat percentage (LMP) and fat meat percentage (FMP) (P〈0.05), while the D103 A744C polymorphism (AY533208) was not significantly associated with any carcass traits. These results indicate that the imprinting status of RTL1 and DIO3 is well kept across the mammalian species, and porcine RTL1 may have important roles in muscle growth and fat deposition.
基金supported by the Major Research Plan of National Natural Science Foundation of China(NO.31690093)Young Elite Scientist Sponsorship Program by CAST(China Association for Science and Technology)
文摘Background: RING H2 finger E3 ligase (RH2FE3) genes encode cysteine rich proteins that mediate E3 ubiquitin ligase activity and degrade target substrates. The roles of these genes in plant responses to phytohormones and abiotic stresses are well documented in various species, but their roles in cotton fiber development are poorly understood. To date, genome wide identification and expression analyses of Gossypium hirsutum RH2FE3 genes have not been reported. Methods: We performed computational identification, structural and phylogenetic analyses, chromosomal distribution analysis and estimated KJKs values of G hirsutum RH2FE3 genes. Orthologous and paralogous gene pairs were identified by all versus all BLASTP searches. We predicted cis regulatory elements and analyzed microarray data sets to generate heatmaps at different development stages. Tissue specific expression in cotton fiber, and hormonal and abiotic stress responses were determined by quantitative real time polymerase chain reaction (qRT PCR) analysis. Results: We investigated 140 G hirsutum, 80 G. orboreum, and evolutionary mechanisms and compared them with orthologs 89 G. roimondii putative RH2FB genes and their in Arobidopsis and rice. A domain based analysis of the G hirsutum RH2FE3 genes predicted conserved signature motifs and gene structures. Chromosomal localization showed the genes were distributed across all G hirsutum chromosomes, and 60 duplication events (4 tandem and 56 segmental duplications) and 98 orthologs were detected, cis elements were detected in the promoter regions of G hirsutum RH2FE3 genes. Microarray data and qRT PCR analyses showed that G hirsutum RH2FE3 genes were strongly correlated with cotton fiber development. Additionally, almost all the (brassinolide, gibberellic acid (GA), indole 3-acetic acid drought, and salt). dentified genes were up regulated in response to phytohormones (IAA), and salicylic acid (SA)) and abiotic stresses (cold, heat, Conclusions: The genome wide identification, comprehensive analysis, and characterization of conserved domains and gene structures, as well as phylogenetic analysis, cis element prediction, and expression profile analysis of G hirsutum RH2FE3 genes and their roles in cotton fiber development and responses to plant hormones and abiotic stresses are reported here for the first time. Our findings will contribute to the genome wide analysis of putative RH2FE3 genes in other species and lay a foundation for future physiological and functional research on G hirsutum RH2FE3 genes.
文摘Objective: To investigate whether deletion of chromosome 3 is involved in the carcinogenesis of primary glioblastoma multiforme (GBM) and to localize the possible common deletion region in the aforementioned chromosome. Methods: PCR based microsatellite polymorphism analyses were performed to detect loss of heterozygosity (LOH). Twenty-three loci on chromosome 3 were examined in 20 cases of GBM. Fluorescence-labeled primers and Perkin Elmer 377 DNA Sequencer were applied. Results: 50% informative cases of GBM displayed LOH on chromosome 3. 50% of informative cases displayed LOH on 3q and 35% on 3p. 25.6% of informative loci showed LOH in our series, in which frequent LOH were observed in the chromosomal region from loci D3S1614 (42.9%) to D3S1565 (35.3%) on 3q24–27 and at loci D3S1569 (35.3%) on 3q22–23 and D3S1289 (33.3%) on 3p14.1–14.3. Conclusion: Loss of genetic material on chromosome 3 may play an important part in the tumorigenesis of GBM. The chromosomal regions from loci D3S1614 to D3S1565 on 3q24–27 and at loci D3S1569 on 3q22–23 and D3S1289 on 3p14.1–14.3 are potential sites for novel tumor suppressor genes associated with GBM.
基金Supported by National Key R&D Projects(2017YFD0201200)Heilongjiang Provincial National Science Foundation(C2016025)
文摘Vegetative Insecticidal Proteins(VIPs), a large family of insecticidal proteins, are produced from Bacillus thuringiensis(Bt) during the vegetative growth stage. VIPs represent the second generation of bio-insecticides that confer a wider insecticidal spectrum and have stronger activity. This work compared the geographical distribution of Bt strains and their vip3 genes in different climatic zones in China, the tropical(Hainan Province), subtropical(Guangxi Province) and temperate zones(Heilongjiang Province). A total of 156 Bt strains were isolated from 841 soil samples in Hainan Province tropical region, 356 Bt strains from 1 420 soil samples in Guangxi Province and 167 Bt strains from 1 010 soil samples in different geographical regions in Heilongjiang Province. Twenty-two out of 156 strains from tropical Hainan Province and two out of 356 from subtropical Guangxi Province were found to express vip3 genes, while vip3 genes were not expressed from temperate zone in Heilongjiang Province. Restriction Fragment Length Polymorphism(RFLP) was used to identify different types of vip3 genes that were within the same family and three fulllength vip3 genes were isolated. The genes cloned from Bacillus thuringiensis strain SL3 expressed in the transformed E. coli BL21 strain. Through SDS-PAGE, 88.6 ku insecticidal protein was expressed. The bioassays used two-instar larva of Lepidoptera insects(Spodoptera exigua and Agrotis ipsilon) were performed. The results of the bioassays showed that the protein strongly inhibited the body weight increasement on Spodoptera exigua and Agrotis ipsilon in a standard bioassay. Taken together, the results indicated that the distribution of Bt strains and vip3 genes had regional preference. Tropical and subtropical regions were the rich resources of Bt strains and vip3 genes compared with temperate region. These results would undoubtedly facilitate the studies of insecticidal proteins and expand the list of the pest-killing candidates to make fully use of the extremely rich microbial resources. The new vip3 genes isolated in the current study might also help resolve the emerging insecticidal resistance problems.
基金National Nature Science Foundation of China(No.82060456)Hainan Provincial Key Research and Development Program Project Fund 405(No.ZDYF2019129)Hainan Provincial Postgraduate Innovation Research Project Fund(No.Hys2019-312)。
文摘Objective:To investigate the core target genes of miR-29b-3p,and analyze the clinical significance of the core target genes in glioma.Methods:Bioinformatics analysis was used to predict and screen the target genes of miR-29b-3p.STRING and Cytoscape software were used to analyze the protein-protein interaction(PPI)of target genes.the differences expression and survival prognosis in glioma were analyzed by GEPIA and CGGA.Independent prognostic factors analyzed by univariate and multivariate Cox proportional hazards regression model.Results:22 target genes of miR-29b-3p were predicted using LinkedOmics,miRDB,miRTarBase,TargetScan,and starbase databases.Through the construction of the PPI network,genes out of the network were removed,and a total of 16 genes were screened for further study of their clinical significance.Based on analysis of GEPIA and CGGA databases,COL2A1,DNMT3A,and DNMT3B were excluded.Through further analysis of the univariate and multivariate Cox proportional hazard regression model,finally identified three core target genes:SERPINH1,LOXL2,CDK6.Conclusion:Bioinformatics analysis showed that miR-29b-3p targeted three core genes such as SERPINH1,LOXL2,and CDK6 in glioma.The expression of these genes was different between brain normal tissues and gliomas,between different grades of tumor,IDH mutation status and 1p/19q codeletion status.Its high expression had adverse effects on overall survival and recurrence-free survival.These core target genes can be used as an independent prognostic factor.