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The T3SS is required for the virulence of many Gram-negative bacterial pathogens of plants and animals. This study focuses on the sensing and function of the T3SS protein HrpP during plant interactions. Our findings show that HrpP and its N-terminal truncation HrpP 1–119 can interact with MKK2, promote effector translocation, and manipulate plant immunity to facilitate bacterial infection, highlighting the P. syringae T3SS component involved in the fine-tuning of plant immunity.

Background Plants and their associated microbiota constitute an assemblage of species known as holobionts. The plant seed microbiome plays an important role in nutrient uptake and stress attenuation. However, the core vertically transmitted endophytes remain largely unexplored. Results To gain valuable insights into the vertical transmission of rice seed core endophytes, we conducted a large-scale analysis of the microbiomes of two generations of six different rice varieties from five microhabitats (bulk soil, rhizosphere, root, stem, and seed) from four geographic locations. We showed that the microhabitat rather than the geographic location and rice variety was the primary driver of the rice microbiome assemblage. The diversity and network complexity of the rice-associated microbiome decreased steadily from far to near the roots, rice exterior to interior, and from belowground to aboveground niches. Remarkably, the microbiomes of the roots, stems, and seeds of the rice interior compartments were not greatly influenced by the external environment. The core bacterial endophytes of rice were primarily comprised of 14 amplicon sequence variants (ASVs), 10 of which, especially ASV_2 (Pantoea) and ASV_48 (Xanthomonas), were identified as potentially vertically transmitted taxa because they existed across generations, were rarely present in exterior rice microhabitats, and were frequently isolated from rice seeds. The genome sequences of Pantoea and Xanthomonas isolated from the parental and offspring seeds showed a high degree of average nucleotide and core protein identity, indicating vertical transmission of seed endophytes across generations. In silico prediction indicated that the seed endophytes Pantoea and Xanthomonas possessed streamlined genomes with short lengths, low-complexity metabolism, and various plant growth-promoting traits. We also found that all strains of Pantoea and Xanthomonas exhibited cellulase activity and produced indole-3-acetic acid. However, most strains exhibited insignificant antagonism to the major pathogens of rice, such as Magnaporthe oryzae and X. oryzae pv. oryzae. Conclusion Overall, our study revealed that microhabitats, rather than site-specific environmental factors or host varieties, shape the rice microbiome. We discovered the vertically transmitted profiles and keystone taxa of the rice microbiome, which led to the isolation of culturable seed endophytes and investigation of their potential roles in plant-microbiome interactions. Our results provide insights on vertically transmitted microbiota and suggest new avenues for improving plant fitness via the manipulation of seed-associated microbiomes.

Strain S150 was isolated from the tobacco rhizosphere as a plant growth‐promoting rhizobacterium. It increased plant fresh weight significantly and lateral root development, and it antagonized plant pathogenic fungi but not phytobacteria. Further tests showed that strain S150 solubilized organic phosphate and produced ammonia, siderophore, protease, amylase, and cellulase, but it did not produce indole‐3‐acetic acid. Using morphology, physiological characteristics, and multi‐locus sequence analysis, strain S150 was identified as Pseudomonas koreensis. The complete genome of strain S150 was sequenced, and it showed a single circular chromosome of 6,304,843 bp with a 61.09% G + C content. The bacterial genome contained 5,454 predicted genes that occupied 87.7% of the genome. Venn diagrams of the identified orthologous clusters of P. koreensis S150 with the other three sequenced P. koreensis strains revealed up to 4,167 homologous gene clusters that were shared among them, and 21 orthologous clusters were only present in the genome of strain S150. Genome mining of the bacterium P. koreensis S150 showed that the strain possessed 10 biosynthetic gene clusters for secondary metabolites, which included four clusters of non‐ribosomal peptide synthetases (NRPSs) involved in the biosynthesis of cyclic lipopeptides (CLPs). One of the NRPSs possibly encoded lokisin, a cyclic lipopeptide produced by fluorescent Pseudomonas. Genomic mutation of the lokA gene, which is one of the three structural NRPS genes for lokisin in strain S150, led to a deficiency in fungal antagonism that could be restored fully by gene complementation. The results suggested that P. koreensis S150 is a novel plant growth‐promoting agent with specific cyclic lipopeptides and contains a lokisin‐encoding gene cluster that is dominant against plant fungal pathogens., We reported a newly isolated Pseudomonas koreensis strain S150, which antagonized plant pathogens, solubilized phosphates, produced siderophores and enzymes, and promoted plant growth. Genome mining showed that the strain S150 possesses 10 genomic islands coding for secondary metabolites, one of which is lokisin, an unusual cyclic lipopeptide. We generated a deletion mutant for the smallest non‐ribosomal peptide synthetase gene lokA in strain S150. Antifungal activity was deficient in the lokA mutant and was restored by gene complementation.

The type III secretion system (T3SS) in many Gram-negative bacterial pathogens is regarded as the most critical virulence determinant and an attractive target for novel anti-virulence drugs. In this study, we constructed a T3SS secretion reporter containing the β-lactamase gene fused with a signal peptide sequence of the T3SS effector gene, and established a high-throughput screening system for T3SS inhibitors in the plant pathogenic bacterium Acidovorax citrulli. From a library of 12,000 chemical compounds, we identified a series of benzyloxy carbonimidoyl dicyanide (BCD) derivatives that effectively blocked T3SS-dependent β-lactamase secretion. Substitution of halogens or nitro groups at the para-position on the benzene ring contributed to an increased inhibitory activity. One representative compound, BCD03 (3,4-dichloro-benzyloxy carbonimidoyl dicyanide), dramatically reduced pathogenicity of A. citrulli on melon seedlings, and attenuated hypersensitive responses in the non-host Nicotiana tabacum caused by pathogenic bacteria A. citrulli, Xanthomonas oryzae pv. oryzae and Pseudomonas syringae pv. tomato at sub-MIC concentrations. Western blotting assay further confirmed that BCD03 inhibited effector secretion from the above bacteria via T3SS in the liquid medium. Taken together, our data suggest that BCD derivatives act as novel inhibitors of T3SS in multiple plant bacterial pathogens.

The aim of the present study was to investigate the effects of baicalein on the protein expression of SATB1 in the MDA-MB-231 human breast cancer cell line. MDA-MB-231 cells were treated with various concentrations of baicalein (0, 10, 20, 40 µM). Following treatment, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and wound healing assay were used to detect the changes in cell proliferation and migration. In addition, western blot analysis was performed to detect the changes in the protein expression levels of SATB1 in the MDA-MB-231 cell line. With the prolongation of administration time and the increase in drug concentration, the inhibitory effect of baicalein on the proliferation and migration of MDA-MB-231 cells gradually increased in a time- and dose-dependent manner (P

Aiming at nonlinearity in indexes of the combat capability system, based on the comprehensive evaluation of the analysis method of system reliability calculation, improve the system of nonlinear characteristics of structure reliability expression. The method is effective using the example of operation capability of certain reconnaissance intelligence armament system.

The Bayesian network for survivability assessment modeling of command information system is introduced. We establish survivability evaluation model from three aspects, the network node, the network structure and model parameters. Proposed the attribute dependence progressive genetic.

microRNAs (miRNAs) act as a major regulator of acquired chemo-resistance in various types of cancer therapeutics. This study investigated the contribution of miRNAs in influencing multiple drug resistance in esophageal squamous cell carcinoma (ESCC). The sensitivity of four ESCC cell lines (EC109, EC9706, TE-1 and KYSE-150) to 5-fluorouracil (5-FU) and oxaliplatin (OX) was determined by MTT assay. A 5-FU and OX-resistant subline, EC9706R, was established by continuous exposure to stepwise increasing concentration of 5-FU and OX. Microarray technology was used to compare the differential expression of miRNAs between resistant cells and parental cells. Chemo-sensitivity assay was performed to evaluate drug response in EC9706R cells transfected with miRNA mimic or inhibitor. The direct targets of miRNA were identified by employing pathway analysis and then confirmed with luciferase assay. Sixty ESCC tissue samples and their paired adjacent normal tissues were collected to validate the expression of identified miRNA. Mouse models were further utilized to investigate the function of miRNA on acquired chemo-resistance. MicroRNA panel results indicated that a total of 12 miRNAs were differentially expressed and miR-141-3p was highly over expressed in resistant cells. Inhibition of miR-141-3p reversed acquired chemo-resistance in EC9706R cells by stimulating apoptosis. The expression of miR-141-3p was significantly increased in ESCC tissue samples compared to their matched distant normal tissues. In addition, the elevated miR-141-3p expression was found to be associated with ESCC differentiation status and TNM stage. Moreover, Phosphatase and tensin homolog (PTEN) was identified as direct target of miR-141-3p. Western blot exhibited altered protein levels of PTEN, Akt, and PI3k with miR-141-3p inhibitor. An inverse correlation between PTEN expression and miR-141-3p expression was also observed in tissue samples. EC9706R xenograft mouse model became sensitized to 5-FU and OX treatment following miR-141-3p inhibitor transfection in vivo. Our study demonstrated that miR-141-3p contributed to an acquired chemo-resistance through PTEN modulation both in vitro and in vivo.

Based on the characteristics of command information system, the concept of survivability is presented. The system survivability framework, analysis model and validation processes are described also.

To explore the expression and significance of respiratory chain enzyme of cells in urine sediment in mitochondrial encephalopathy myopathy, lactic acidosis and stroke-like episodes (MELAS) syndrome.Through enzyme histochemistry, the authors analyzed the changes of respiratory chain enzyme in urine sediment in 20 MELAS patients due to mitochondrial A3243G mutation (MELAS group) and 20 health peoples (control group). And the impact on the expression of protein encoded by nuclear DNA (A21347) and mitochondrial DNA (A6404) was detected by immunochemistry. Image pro Plus 6.0 software was used for analysis of absorbance (A) of staining images as staining intensity. The data were expressed as M (Q1, Q3) and analyzed through statistical software.The staining intensity of complexes Iin the MELAS group was lower than that in the control group (0.06(0.01, 0.12) vs 0.12(0.01, 0.62), P = 0.010). The intergroup staining intensity of complex II showed no marked difference. Increased density of blue particle and cytoplasmic gathering was found in 13 cased (65%) of the MELAS group under light microscope. The staining intensity of complexes IV was expressed at a low level in the MELAS group (0.14(0.03, 0.32) vs 0.23(0.06, 0.43), P = 0.038). The expression of protein encoded by nuclear DNA (A21347) was lower than that in the control group (0.05(0.02, 0.45) vs 0.17(0.03, 0.70), P = 0.000). The expression of protein encoded by mitochondrial DNA (A6404) was also lower than that in the control group (0.03(0.01, 0.07) vs 0.15 (0.09, 0.23), P = 0.000).Abnormal change of respiratory chain enzyme in urine sediment in MELAS due to mitochondrial A3243G mutation and a low expression of proteins encoded by two kinds of DNA in complexes IV can help to confirm the genetic diagnosis of mitochondrial encephalomyopathies so that different subtypes may be classified and its pathogenesis elucidated.

To summarize the clinical audiologic features of patients with mitochondrial DNA (mtDNA) A3243G mutation and explore the lesion location of hearing loss so as to examine its correlation with the related syndrome.A total of 44 patients with mtDNA A3243G mutation from 2009-2011 were studied. Audiological evaluations consisted of measurements of pure-tone and speech audiometry, tympanometry, distortion-product otoacoustic emissions and auditory brainstem response. We investigated a possible correlation between the degree of hearing loss and gender, age and mutation rate.(1) Pure tone test was performed in 41 patients and showed normal hearing or symmetrical sensorineural hearing loss. Pure tone audiogram (PTA) showed high-frequency loss and descending curve in a majority of patients. There were 75 ears with hearing loss in 82 ears (91.46%), 22 ears with abnormal speech audiometry in 26 ears, 77 ears with abnormal distortion product otoacoustic emissions (DPOAE)testing in 86 ears, including 5 ears with normal PTA, 31 ears with abnormal electrocochleography in 75 ears, 25 ears with abnormal auditory brainstem response (ABR) in 82 ears. The abnormal ABR showed elevated threshold in 10 ears, delayed interpeak latencies of wave I-V in 2 ears and disappearance of wave V before wave I in 1 ear. In addition, there were 2 ears with speech audiometry abnormal but with normal ABR. (2) The correlation between the severity of hearing and gender did not reach statistical significance, nor the severity of hearing and mutation ratio. Age could influence the hearing of A3243G-induced MELAS.The predominant lesions of mtDNA A3243G is at cochlea and retrocochlear sites. Significant variations in clinical manifestation of hearing are the prominent features in patient with A3243G mutation. There was no correlation between the degree of hearing loss and mutation load. However, hearing impairment is the most common symptom of A3243G mutation.

In order to summarize and analyze the progresses and problems of resistance testing and technology research on acupoints, investigate the related solutions, articles on resistance testing from 1990 to 2011 were collected, and advantages and shortcomings of different types of testing instruments were discussed in this article as well. Detection technology of acupoint impedance has under updating changes, which are often applied on locating acupoints and clinical diagnosis. The progresses of those technologies are mainly embodied in improvement of circuit model, which enable a longer testing duration, less media interference and so on. The previous studies often focus on resistance testing of the skin. Since dermal resistance does not equal to acupoint resistance, the designation of equivalent circuit does not embody the condition of cells, tissues and nerves at acupoints, and testing technology is often affected by factors such as polarization of electrodes, contact medium and anatomical characteristics of acupoints, the resistance of acupoints can not be taken as the effective framework of the diagnostic standard and judgement of therapeutic effect in clinic. Based on the key problems in resistance testing of acupoints, it is proposed by the article that the techniques and designation of circuit model should be reformed, and mathematical model should be established in the study to meet the requirement of the life system. Inductance of human body should be considered in resistance research of acupoints, and interference factors should be eliminate as far as possible in designation of equipments. Thus, the change law of acupoint resistance in the inner part of the body can be fully embodied by the designation of experiments.

Aggregation of systematic operational capability of armaments is significant for armaments systems' developing, constructing and using. As holistic avalanche and operational capability's nonlinear character of armaments systems, the feedback mechanism among capability indexes is researched and an aggregate method of systematic operational capability based on feedback mechanism is proposed on this paper, what's more, the method is proved effective by certain example of reconnaissance intelligence armaments systems' operational capability aggregation.

To investigate the spectrum of mitochondrial DNA (deoxyribonucleic acid) 3271TC, 8356TC, 9176TC/G and 13513GA mutations in Chinese patients with mitochondrial encephalomyopathies.Peripheral blood samples were collected from 500 mitochondrial encephalomyopathies patients clinically diagnosed as mitochondrial encephalomyopathy lactic acidosisstroke-like episodes (MELAS), myoclonus epilepsyragged-red fibers (MERRF) or Leigh's syndrome from October 2005 to October 2009. The methods of PCR- polymerase chain reaction-restriction fragment length polymorphism (RFLP) and PCR-sequencing were performed to identify the mutations.No patients with the 3271TC, 8356TC, 9176TC/G or 13513GA mutations were identified.The mutations of 3271TC, 8356TC, 9176TC/G and 13513GA are rare causes of mitochondrial encephalomyopathies in Chinese patients.

To understand the clinical and genetic features of Huntington disease (HD).The clinical data of HD cases from 2 Chinese families were analyzed and trinucleotide repeat in the IT15 gene were investigated in 9 of the two families by polymerase chain reaction and GeneScan.Among the two pedigrees, 6 cases were ascertained as HD by genetic test. Genotypes of IT15 were heterozygous in these HD patients. CAG repeat of the patients in the HD chromosome were 40-78. In the two pedigrees, the onset age was earlier in the subsequent generations than that of their fathers. In pedigree 2, the onset age was inversely correlated with CAG repeat number. One out of the 6 cases was juvenile-onset type of Huntington disease, whose clinical symptoms were different from those of the adult-onset cases, especially the hypertonic manifestation.HD is an autosomal dominant neurodegenerative disorder with genetic anticipation caused by enlargement of CAG repeat in IT15 gene. The clinical manifestation is different between the juvenile-onset and the adult-onset. The number of CAG repeat is inversely correlated with the onset age and clinical severity.

A3243G mutation in mitochondrial DNA is the most common pathogenic point mutation causing a variety of phenotypes. The clinical phenotype and the relationship between the clinical phenotype and the ratio of A3243G mutation were studied in the members from nuclear families carrying A3243G mutation.A total of 42 families carrying A3243G mutation were recruited and their clinical symptoms, laboratory results and the ratio of A3243G analyzed.(1) In probands, myopathy, seizure, hirsutism, headache, cognitive impairment, weight loss and short stature were the most common clinical features. They tended to occur simultaneously. Lactic acid, pyruvate and MRI were abnormal in most probands; (2) most carriers had a normal phenotype. Myopathy, weight loss and short stature were their most common clinical features; (3) the ratio of A3243G mutation in urine was higher than that in blood in probands (t = -15.06, P0.001). And the ratio of A3243G mutation in urine was higher than that in blood in their mothers (z = -6.241, P0.001); (4) the ratio of A3243G mutation in probands was 2-fold higher than that in their mothers in both blood and urine.The phenotype of patients carrying A3243G mutation is varied. The clinical symptoms and laboratory results of probands are worse than those of mothers. It is probably due to a higher mutation ratio of m.3243AG in their tissues.

To explore effects of 650 nm laser and moxibustion pretreatment on visceral traction pain (VTP) and its mechanism.Forty male SD rats were randomly devided into a sham operation group (group A), a VTP group (group B), a 650 nm laser pretreatment group (group C), a moxibustion pretreatment group (group D). Rats in group A and group B were not treated except sham operation or VTP model. In group C and D, the VTP models were produced immediate after 650 nm laser irradiation or moxibustion at "Zusanli" (ST 36), respectively. The changes of pain score and systolic pressure were investigated and the activity of AChE, the content of SP and leu-enkephaline (LEK), and the positive index of c-Fos protein and glial fibrillary acidic protein (GFAP) were detected by biochemistry, radio-immunity method and immunohistochemistry, respectively.Compared with group A, the pain score, systolic pressure, the activity of AChE, the content of SP, and the positive index of c-Fos protein and GFAP of group B increased significantly (all P0.05); compared with group B, the pain score, AChE activity, the content of SP and the positive index of c-Fos protein and GFAP of both group C and group D decreased significantly (all P0.05); compared with group B, the content of LEK increased and systolic pressure decreased significantly in group C (both P0.05).Both 650 nm laser and moxibustion pretreatment can inhibit VTP and the mechanism may be related to reducing the activity of AChE and the content of SP, and increasing the activity of LEK and decreasing the expression of c-Fos protein and GFAP.

To identify a better non-invasive method to detect the carrier of mitochondrial A3243G mutation, a cause of mitochondrial encephalopathy-lactic acidosis-stroke like episode (MELAS) syndrome.DNA was extracted from the peripheral blood, urine, hair follicle, and saliva of 25 MELAS syndrome patients carrying A3243G mutation and their mothers and other maternal relatives, 33 persons in number, and the muscle tissues from 5 patients obtained by biopsy. A3243G mutation was detected by PCR-RFLP method, and the A3243G mutation ratio was identified by measuring the density of each band and calculation with the software AlphaEase 5.0.A3243G mutations were detected in all tissues of the 25 MELAS patients. The A3243G mutation ratio in urine was 62% +/- 9%, significantly higher than that in the blood [(36% +/- 10%), t = -11.13, P0.01]. A3243G mutations were detected in at least one tissue of the 28 maternal relatives. The A3243G mutation rates in their urine samples was 33.0% (5.0% - 70.4%), significantly higher than that in their blood samples [8.0% (0 - 33.3%), z = -4.197, P0.01]. There was no significant difference in A3243G mutation ratio among the samples of hair follicle, saliva, and blood.The A3243G mutation ratio in urine is significantly higher than those in blood samples of the patients and their maternal relatives. A noninvasive method, A3243G mutation ratio analysis of urine is superior to that in blood.

To conduct a molecular epidemiological survey on the mitochondrial DNA C1494T mutation in non-syndromic hearing loss patients in Chinese population.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to screen the mitochondrial DNA 12S rRNA C1494T mutation in 20 patients with aminoglycoside antibiotic induced hearing loss, 136 sporadic non-syndromic hearing loss patients and 50 probands of pedigrees with non-syndromic hearing loss.The C1494T mutation did not appear in all cases except for the positive control.Incidence of mitochondrial DNA C1494T mutation is much lower than that of mitochondrial DNA A1555G mutation in non-syndromic hearing loss of Chinese population. Mitochondrial DNA C1494T mutation may be a rare variation in non-syndromic hearing loss and is not the main cause of aminoglycoside antibiotic induced-deafness.

To investigate the association between single nucleotide polymorphisms (SNPs) of casein kinase I gamma 2 (CSNK1G2) gene and children with familial febrile convulsions.The study samples were collected from unrelated Chinese Han population of Hebei province, including a cohort of 53 children with familial febrile convulsions(FC) and a control cohort of 101 individuals. Genotypes of SNPs rs2074882, rs740423, rs2277737, rs4806825, rs1059684 were typed by polymerase chain reaction-restriction fragment length polymorphism.The frequencies of the five SNPs complied well with the Hardy-Weinberg equilibrium in FC group and normal group. The distribution of genotype and frequencies of alleles of the SNPs rs740423, rs2277737, rs1059684 in familial febrile convulsions group was significantly different from that in control group. No significant difference was observed in the distribution of genotypes and frequencies of alleles at SNP rs2074882 between two groups. Analysis on rs4806825 was not made owing to its less allele frequency.These data indicate that SNPs rs740423, rs2277737, rs1059684 of CSNK1G2 gene may contribute to familial febrile convulsions in children.